Manual Software ASpect UV en

Operating manual
ASpect UV
Software for SPECORD PLUS
Manufacturer Analytik Jena GmbH
Konrad-Zuse-Str.1
07745 Jena  Germany
Phone + 49 3641 77 70
Fax + 49 3641 77 9279
Email info@analytik-jena.com
Service Analytik Jena GmbH

Konrad-Zuse-Str. 1
07745 Jena  Germany
Phone + 49 3641 77 7407
Email service@analytik-jena.com
General information http://www.analytik-jena.com
Documentation number 10-2210-002-23
Edition A (01/2021)
Implementation of the Analytik Jena GmbH

Technical Documentation
© Copyright 2021, Analytik Jena GmbH

Inhalt
Inhalt
1 About this document .................................................................................................. 7
2 Install ASpect UV ........................................................................................................ 8
3 Starting and exiting ASpect UV .................................................................................. 9

3.1 Starting ASpect UV ............................................................................................................ 9
3.2 Starting ASpect UV with a module or quick start ............................................................. 9
3.3 Exiting ASpect UV ............................................................................................................ 10
4 What you should know about ASpect in advance .................................................... 11

4.1 ASpect UV program structure ......................................................................................... 11
4.1.1 Modules in ASpect UV ..................................................................................................... 11
4.1.2 The main window in ASpect UV...................................................................................... 12
4.2 The document window ................................................................................................... 15
4.2.1 Overview of the document window ................................................................................ 16
4.2.2 Menu bar and File menu in the document window ....................................................... 17
4.2.3 Worksheets in the document .......................................................................................... 18
4.2.4 Show spectra and measurement curves on the worksheet ........................................... 19
4.3 Managing, printing and exporting documents ............................................................. 22
4.3.1 Opening and saving documents ..................................................................................... 23
4.3.2 Creating a new document ............................................................................................... 24
4.3.3 Editing document titles ................................................................................................... 25
4.3.4 Tracking changes in the audit trail ................................................................................. 25
4.3.5 Print documents .............................................................................................................. 26
4.3.6 Export data ...................................................................................................................... 29
4.3.7 Automatically archiving documents ............................................................................... 30
4.4 Sample sequences ........................................................................................................... 32
4.4.1 Structure of sample sequence and sample types ........................................................... 32
4.4.2 Create sample sequence ................................................................................................. 34
4.4.3 Edit sample sequences .................................................................................................... 37
4.5 Using formulas ................................................................................................................ 38
4.6 Using accessories in SPECORD PLUS .............................................................................. 40
4.6.1 Passive accessories .......................................................................................................... 40
4.6.2 Cassette sipper system .................................................................................................... 40
4.6.3 Sampler ........................................................................................................................... 41
4.6.4 Cell changer ..................................................................................................................... 42
4.6.5 Peltier-tempered accessories .......................................................................................... 43
4.6.6 Integrating sphere ........................................................................................................... 47
4.6.7 Reflectance measuring attachments .............................................................................. 47
4.6.8 Scanning attachment for solid samples ......................................................................... 48
5 Quick measurement module and online measurements ......................................... 49
6 Photometry module ................................................................................................. 52

6.1 Method configurations in Photometry module.............................................................. 52
6.1.1 Photometry – Settings / General .................................................................................... 52
6.1.2 Photometry – Settings / Accessory................................................................................. 54
6.1.3 Photometry – Settings / Evaluation ............................................................................... 55
6.1.4 Photometry – Settings / Calibration ............................................................................... 55
6.1.5 Photometry – Settings / Sample sequence .................................................................... 56
6.1.6 Photometry – Settings / Archive automatically ............................................................. 56
6.1.7 Photometry – Settings / Notice ...................................................................................... 57
6.2 Performing measurements in Photometry module ....................................................... 57
6.3 Displaying, analyzing and editing results in the Photometry module ........................... 59
6.3.1 Edit sample table in Photometry module ....................................................................... 61
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6.3.2 Analyzing photometry data............................................................................................. 62

6.3.3 Editing photometry data ................................................................................................. 65
6.4 Example measurement for the Photometry module...................................................... 67
7 Spectrum module ..................................................................................................... 71

7.1 Method configuration in the Spectrum module ............................................................. 71
7.1.1 Spectrum - Settings / General......................................................................................... 72
7.1.2 Spectrum – Settings / Accessory ..................................................................................... 73
7.1.3 Spectrum – Settings / Evaluation ................................................................................... 73
7.1.4 Spectrum – Settings / Calibration ................................................................................... 75
7.1.5 Spectrum – Settings / Sample sequence ........................................................................ 76
7.1.6 Spectrum – Settings / Archive automatically ................................................................. 76
7.1.7 Spectrum – Settings / Note............................................................................................. 76
7.2 Performing measurements in the Spectrum module ..................................................... 77
7.3 Displaying, analyzing and editing results in the Spectrum module ............................... 79
7.3.1 Display options for the results in the Spectrum module ................................................ 80
7.3.2 Analyzing spectral data ................................................................................................... 80
7.3.3 Editing spectral data........................................................................................................ 82
7.3.4 Combining measuring results from different documents .............................................. 87
7.4 Example measurement for the Spectrum module ......................................................... 87
8 Kinetics module ........................................................................................................ 91

8.1 Method configuration in the Kinetics module ................................................................ 91
8.1.1 Kinetics - Settings / General ........................................................................................... 91
8.1.2 Kinetics – Settings / Accessory........................................................................................ 93
8.1.3 Kinetics – Settings / Evaluation ...................................................................................... 93
8.1.4 Kinetics – Settings / Calibration...................................................................................... 94
8.1.5 Kinetics – Settings / Sample Sequence........................................................................... 96
8.1.6 Kinetics – Settings / Archive automatically .................................................................... 96
8.1.7 Kinetics - Setting / Note .................................................................................................. 96
8.2 Performing measurements in the Kinetics module ........................................................ 96
8.3 Displaying, analyzing and editing results in the Kinetics module .................................. 98
8.3.1 Display options for the results in the Kinetics module ................................................. 100
8.3.2 Evaluate kinetic curves .................................................................................................. 100
8.3.3 Editing kinetics data ...................................................................................................... 103
8.4 Example measurement for the Kinetics module .......................................................... 104
9 Thermometry module ............................................................................................ 108

9.1 Method configuration in the Thermometry module .................................................... 108
9.1.1 Thermometry – Settings / General ............................................................................... 108
9.1.2 Thermometry – Settings / Accessory ............................................................................ 110
9.1.3 Thermometry – Settings / Evaluation .......................................................................... 112
9.1.4 Thermometry – Settings / Sample Sequence ............................................................... 113
9.1.5 Thermometry – Settings / Archive automatically ........................................................ 113
9.1.6 Thermometry – Settings / Note.................................................................................... 113
9.2 Performing measurements in the Thermometry module ............................................ 114
9.3 Displaying, analyzing and editing results in the Thermometry module ...................... 116
9.3.1 Display options for the results in the Thermometry module ....................................... 117
9.3.2 Evaluating the temperature curves ............................................................................... 117
9.3.3 Editing temperature curves ........................................................................................... 119
9.4 Example measurement for the Thermometry module ................................................ 121
10 Colorimetry module................................................................................................ 126

10.1 Method configuration in the Colorimetry module ....................................................... 126
10.1.1 Colorimetry - Settings / General ................................................................................... 127
10.1.2 Colorimetry – Settings / Accessory ............................................................................... 128
10.1.3 Colorimetry – Settings / Color values ........................................................................... 128
10.1.4 Colorimetry – Settings / Sample Sequence .................................................................. 130
10.1.5 Colorimetry – Settings / Archive automatically ............................................................ 130
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10.1.6 Colorimetry – Settings / Note ....................................................................................... 130

10.2 Performing measurements in the Colorimetry module ............................................... 130
10.3 Displaying, analyzing and editing results in the Colorimetry module ......................... 132
10.3.1 Display options for the spectra in the Colorimetry module ......................................... 134
10.3.2 Analyzing spectra in the Colorimetry module .............................................................. 135
10.4 Exemplary measurement in the Colorimetry module .................................................. 135
11 General functions of the SPECORD PLUS – Menu Device ...................................... 139

11.1 Show device status ........................................................................................................ 139
11.2 Initializing the SPECORD PLUS and starting simulation mode .................................... 139
11.3 Correct SPECORD PLUS ................................................................................................. 140
11.4 Checking the measuring technology condition of the SPECORD PLUS. ...................... 141
11.4.1 Checking the lamp intensity.......................................................................................... 141
11.4.2 Checking the functionality ............................................................................................ 142
11.5 Measuring energy ......................................................................................................... 143
11.6 Setting the zeroth order ................................................................................................ 144
11.7 Additional functions for controlling accessories .......................................................... 144
11.7.1 Switching active accessory detection on and off .......................................................... 144
11.7.2 Switching accessories on and off .................................................................................. 145
11.7.3 Adjusting the cell changer and moving to individual positions ................................... 145
11.7.4 Optimizing the pumping time of the cassette sipper system ...................................... 146
11.7.5 Adjusting the sampler, moving to individual positions and optimizing the pumping time
....................................................................................................................................... 148
12 Setting ASpect UV in the Options window ............................................................. 152

12.1 Configuring the launch bar ........................................................................................... 152
12.2 Configuring quick start .................................................................................................. 153
12.3 Directory path presets ................................................................................................... 154
12.4 Setting the language ..................................................................................................... 154
12.5 Defining the start mode for ASpect UV ........................................................................ 154
12.6 Switching on the lamps and recording monitor files ................................................... 155
12.7 Configuring simulation mode ....................................................................................... 155
12.8 Displaying reference measurements ............................................................................ 156
12.9 Configuring calibration ................................................................................................. 156
12.10 Defining decimal places ................................................................................................ 157
12.11 Configuring password properties .................................................................................. 157
12.12 Creating signatures ....................................................................................................... 158
12.13 Activating automatic archiving ..................................................................................... 159
12.14 Configuring the CSV export ........................................................................................... 160
12.15 Configuring the webserver ............................................................................................ 160
13 Validating the SPECORD PLUS (optional) .............................................................. 161

13.1 Validation AJ .................................................................................................................. 161
13.1.1 Test parameters and required equipment .................................................................... 162
13.1.2 Entering validation parameters .................................................................................... 164
13.1.3 Running the validation ................................................................................................. 167
13.2 Validation Standard Ph.Eur........................................................................................... 169
13.3 Validation according to USP .......................................................................................... 176
13.3.1 Test parameters, acceptance criteria and required test equipment ............................ 177
13.4 Validation Maintenance Economic ............................................................................... 187
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Inhalt
14 User administration and electronic signatures ...................................................... 194

14.1 User management ......................................................................................................... 194
14.1.1 Preinstalled users .......................................................................................................... 194
14.1.2 Functions in the user administration ............................................................................ 196
14.1.3 Create and edit user profiles ......................................................................................... 197
14.1.4 Assign user rights .......................................................................................................... 198
14.1.5 Creating reports on the user management .................................................................. 199
14.1.6 Log off user .................................................................................................................... 200
14.1.7 Lock ASpect UV .............................................................................................................. 200
14.1.8 Passwords in ASpect UV ................................................................................................ 201
14.2 Electronic signatures ..................................................................................................... 201
15 AJ File Protection .................................................................................................... 203
16 WebApp .................................................................................................................. 204

16.1 Sent data........................................................................................................................ 204
16.2 Configuring the network for using the web server ....................................................... 204
16.3 Configuring the instrument list ..................................................................................... 205
16.4 Supported end instruments .......................................................................................... 206
16.5 Opening the WebApp from the end instrument .......................................................... 206
17 Overview of revisions ............................................................................................. 207
18 Supplement ............................................................................................................ 208

18.1 Supplement A - Default methods ................................................................................. 208
18.2 Supplement B - Generation of measured values and recording of reference values .. 211
18.3 Supplement C – Data format for importing sample information ................................ 212
19 Index .......................................................................Fehler! Textmarke nicht definiert.
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About this document
1 About this document

What is ASpect UV The ASpect UV program is used to control and analyze the measured data from the
following devices of Analytik Jena GmbH:
 SPECORD 250 PLUS
 SPECORD 50 PLUS
Program versions This description is based on the following program versions:

 ASpect UV from V. 1.5
 ASpect UV Firmware from 4.4.2
The program version is displayed if you select Info  About ASpect UV from the menu
in the main window.
Target group This documentis aimed at qualified specialist personnel with knowledge of UV/Vis
analysis. Basic knowledge of working with a computer and programs, such as marking
an option by clicking with the mouse or saving and opening files, is assumed.
For the safe operation of the SPECORD PLUS with ASpect UV knowledge of the operating
instructions "SPECORD PLUS UV/Vis - spectral photometer" and " SPECORD PLUS
Accessories UV/Vis spectral photometer " is required.
Typographical Instructions for action are numbered or grouped into blocks of instructions, shown as
conventions bullet points.
The elements of the ASpect UV program are indicated as follows:
 All elements, such as menu items, window names, buttons etc. are indicated with
small caps, e.g. Modules.
 Menu items in a command sequence are separated by an arrow (), e.g. File  Open.
 Buttons are indicated by square brackets, e.g. [OK].
The following pictograms and signal words are used:
Attention This is a note to be followed to avoid operating errors and
obtain correct results.
Tip The notes identified in this manner provide you with useful
information concerning the program control.
7
Install ASpect UV
2 Install ASpect UV
Attention The installation of ASpect UV requires administrator rights for the computer.
For the correct installation of the USB driver and to update the firmware the following
sequence must be adhered to:
 First install the ASpect UV program on the computer.
 Only then connect the SPECORD PLUS to the computer and switch on the SPECORD
PLUS.
When the SPECORD PLUS is connected for the first time, a device recognition procedure
is performed, and the drivers are allocated after the device is switched on.
Install ASpect UV  Switch on computer.

 Insert the ASpect UV installation CD into the CD-ROM drive.
On most computers, the installation will start automatically after a brief delay. If this
does not happen, start the file setup.exe in the top directory of the CD.
 Click on [Install] and follow the instructions of the installation program:
− Select the language of installation.
− Select the target folder for the installation.
 The installation completes automatically.
 Connect the SPECORD PLUS to the computer vis the USB connections.
 Switch on the SPECORD PLUS.
 The required USB drivers will be installed automatically.
Install optional modules The optionally available modules are also supplied on CDs.
 Insert the installation CD into the drive.
If the installation does not start automatically, start the file setup.exe on the CD.
 Install the programs in the same folder as ASpect UV.
 The optional modules are integrated into the program when ASpect UV starts.
8
Starting and exiting ASpect UV
3 Starting and exiting ASpect UV

With the default settings of Aspect UV installed the main window is displayed after
starting the program and the connection to the SPECORD PLUS and connected
accessories is established. However, you may also configure the program in such a way
that an application module opens straight away, or the quick start menu is shown with a
collection of selected methods.
3.1 Starting ASpect UV
By default the connection between the computer and SPECORD PLUS is established
during the start of ASpect UV. The connection sequence must be observed:
Execution
 Install the accessories for the measurement in the sample chamber and switch on
SPECORD PLUS from the mains switch on the right side of the housing.
 Start ASpect UV by clicking on the desktop icon or by selecting Windows Program
icon  ASpect UV  ASpect UV from the Windows desktop.
 Only if using the module ASpect UV FDA 21 CFR Part 11:
− In the login window, enter the Login (user name) and Password.
 ASpect UV starts and establishes the connection to the SPECORD PLUS. The
monochromator of the SPECORD PLUS moves and the message "Initialization" is
shown on the monitor. The SPECORD PLUS is now ready to measure.
Tip
If the connection to SPECORD PLUS could not be established, ASpect UV is started in
simulation mode. Check the settings in the window Options / Start / Start ASpect UV.
There, the checkbox Initialize instrument must be enabled (→ "Defining the start mode
for ASpect UV" p. 154). If necessary, establish the connection later by selecting
Instrument  Initialization from the menu (→ "Initializing the SPECORD PLUS and
starting simulation mode" p. 139).
3.2 Starting ASpect UV with a module or quick start
By default ASpect UV starts with the main window as start screen. However, the
program start can also be configured for a favorite module or the quick start menu to be
displayed.
Execution
 Select Setting  Options from the menu.
Alternatively, click on [Options] in the launch bar of the main window.
 In the window Options / Start / Start ASpect UV, select the desired module or the
quick start from the list Open element. With the option None, ASpect UV starts
with the main window.
9
Starting and exiting ASpect UV
 When selecting the quick start, configure the quick start menu (→ "Configuring
quick start" p. 153).
 During the next start of ASpect UV, the selected element is displayed as start
screen.
3.3 Exiting ASpect UV
With the software-assisted shutdown routine, the monochromator of the spectrometer

is locked into the original position. In this position, the transport lock of the
monochromator can be attached (see SPECORD PLUS UV/Vis Spectral Photometer
operation instructions).
Execution
 Close all document windows.
 Select the File  Close menu command.
 The message "shut-down routine running" is displayed.
 After exiting the ASpect UV program, switch off the SPECORD PLUS from the mains
switch.
 The SPECORD PLUS is shut down.
If the work at the SPECORD PLUS has been completed and only the analysis of the data
in the ASpect UV remains, first switch to the simulation mode (Instrument 
Simulation). The USB connection to the device is thus closed and you can switch off
SPECORD PLUS at the power switch. The ASpect UV program can be ended later.
10
What you should know about ASpect in advance
4 What you should know about ASpect in advance

This chapter includes information about the program structure and cross-module
program functions of ASpect UV.
4.1 ASpect UV program structure
4.1.1 Modules in ASpect UV
The ASpect UV program is of modular design. Each application area of the UV/Vis
spectroscopy has a module assigned to it, in which the measuring parameters to be
configured are adapted to the application and the connected accessories. This permits a
quick orientation when selecting the required analysis parameters and simplifies data
analysis.
The following applications/modules are available:
 Photometry
In this module, measurements are taken at selected wavelengths. Quantitative
analyses at selected wavelengths for routine analysis, as well as all analyses
integrating the measured values at selected wavelength into a formula, e.g.
biochemical analyses, can be implemented.
 Spectrum
This module permits the measuring of spectra or spectral ranges. Based on this
mathematic data processing of the spectra or a peak search can be performed.
Sophisticated quantitative determinations based on the spectral analysis are also
possible (automatic peak search, peak areas, analysis of spectral derivations, etc.).
 Kinetics
This module permits the recording of time-dependent reactions, i.e. registration of
the change of transmission or absorbance at a wavelength. The graph and measured
values for the selected points in time are analyzed.
 Quick measurement
This module permits a quick determination of absorbance at a wavelength. Further
processing, saving or printing of the values is not possible.
 Thermometry
This module is used to evaluate temperature-dependent absorbance or transmission
changes. The temperature curves can be used to determine the DNA melting point.
The temperature curves are recorded by means of Peltier-tempered accessories.
 Colorimetry
The color measurement module determines the color coordinates of reflecting
surfaces or optically transparent samples. Moreover, different color numbers can be
determined.
Optionally the following modules are available:
 ASpect UV Validation AJ
 ASpect UV Validation Standard Ph. Eur.
 ASpect UV Validation USP
11
 ASpect UV Maintenance economic

 AJ File Protection
Also available as an option is the ASpect UV FDA 21 CFR Part 11 software, which
includes user management and file signing in addition to the functions of ASpect UV.
4.1.2 The main window in ASpect UV
The main window opens after starting ASpect UV. It provides the following functions:
 Open the modules with the related measuring and analysis functions
 Configuration of module-independent basic functions for measurement and analysis
(options)
 Functions for testing the SPECORD PLUS
 Access to user administration if the module ASpect UV FDA 21 CFR Part 11 has been
installed
ASpect UV main window
Components in the main The list below includes the key components of the main window. The numbers
window correspond to those in the figure above:
No. Description
1 The launch bar contains the icon buttons for the modules and basic program functions.
2 The general functions of ASpect UV have been arranged in menus of the menu bar.
3 The documents are arranged in the workspace (→ "The document window" p. 15).
Menu bar The menu bar of the main window includes the following functions:
Menu Subitem Description
File  Open method Open a saved method for a subsequent measurement
Open results Open a saved result in its module environment
12
Close Exiting ASpect UV
Settings  Hide menu Hide menu bar
Hide/show Hide launch bar

launchbar
Options Make basic configurations for ASpect UV
Module  Quick Start quick measurements at a selected wavelength

measurement
Spectrum Open a new document in the module Spectrum / Change
to the module Spectrum
Photometry Open a new document in the module Photometry /

Change to the module Photometry
Kinetics Open a new document in the module Kinetics / Change to
the module Kinetics
Colorimetry Open a new document in the module Colorimetry /
Change to the module Colorimetry
Thermometry Open a new document in the module Thermometry /

Change to the module Thermometry
Quick start Open the quick start menu
Instrument Info After initialization, display information on the device, the

 DCU version and the connected accessories
Initialization Establish the connection between SPECORD PLUS and
ASpect UV
Simulation Put ASpect UV in simulation mode
Test Test the functionality of the SPECORD PLUS

Validate SPECORD PLUS (optional modules)
Correction Start automatic software-supported device corrections
Zeroth order Set the monochromator grating to the zero order for
adjusting accessories and testing the beam paths
Accessory Switch the accessory on and off; with a sipper, start the
pump time optimization; with Peltier accessories, query
the temperatures
User Log off Log off the user and lock ASpect UV
Change Change password of logged-on user

password
User Open user administration
management
Info  Help Open help for ASpect UV
About Program version
Launch bar The launch bar on the left side of the main window includes icons providing quick access
to the most frequently used program functions. The launch bar can be configured in the
13
Options menu (→ "Configuring the launch bar" p. 152). The following functions can be
arranged:
Symbol Functional description
Open Photometry module / Add new document
Open Spectrum module / Add new document
Open Kinetics module / Add new document
Open Thermometry module / Add new document
Open Colorimetry module / Add new document
Load saved method for next measurements
Open saved measured values and analyses
Initialize SPECORD PLUS
Display quick start menu with method selection
Open Options window
Exiting ASpect UV
Log off user
Display of the absorbance value at the currently configured wavelength of

the monochromator
Status bar The following information is displayed in the status bar:

 SIM: The software is in simulation mode.
 INIT: The device was initialized at the indicated time.
 Device type and serial number
 Connected accessories
 Module: Measurement is being performed in the indicated module.
 Logged in user
 HL / D2E: Lamp status
14
The lamp status is indicated via colored fields:
Status color Meaning
Gray The lamps are not switched on or the software is in simulation mode.
Green The lamps are in operation.
Red The lamps are defective.
Show and hide the menu If required, the menu and launch bar can be shown and hidden to enlarge the workspace
and launch bar of the main window.
Execution
 To hide the menu, select Settings  Hide menu.
 To show the menu again, click on the dropdown menu icon in the top left corner of
the main window.
Icon for showing the menu in the main window
 The launch bar can be hidden and shown by clicking on Settings  Hide/show
launchbar.
Use help Help on the operation of ASpect UV is available via Info  help. In the ASpect UV
windows, you can activate the [Help] button at the bottom left of the window or the
function key [F1] to obtain information relating to the window settings.
4.2 The document window
ASpect UV uses a single structure for all files, also referred to below as documents, which
includes all method configurations, sample names, measurement data and analyses.
The document window in which a document is opened is of similar design in all
modules. It includes functions for measurements, data administration and analysis. This
section explains the general control elements and functions of the module windows.
Functions that are only available in individual modules are described in the chapters
relating to the corresponding modules.
Change between open If several documents of the same module have been opened, these are arranged behind
documents each other in the form of tabs in the workspace of the main window. Only the
15
documents of one module are displayed simultaneously. If documents of different

modules have been opened, it is necessary to change to the corresponding module by
clicking on the module icon in the launch bar or selecting the menu item in the Module
menu in order to display them.
4.2.1 Overview of the document window
Example of a document window of the Spectrum module
The list below includes the key components of the document window. The numbers
correspond to those in the figure above:
No. Description
1 The module name indicates which module has been opened.
2 The document title is used to differentiate the opened documents. It is defined during
the method configuration and may be later edited. If several documents in a module are
opened, it is possible to change to the corresponding document by clicking on the tab
with the document title.
3 The menu and function bar provides functions for measurements and file
administration in the document.
4 The worksheet includes measured values, analyses, data processing and audit trail.
Several worksheets with different analyses can be created in a document (→
"Worksheets in the document" p. 18).
5 The file name of the document is displayed after saving.
6 The function menu of the worksheet includes the menus for analysis, data processing
and the view of the worksheet.
7 Display of the accessories used in the method. A method is always tailored to an
accessory and can only be used with this accessory.
8 On the tabs, the method parameters of the methods saved in the document, the note
relating to the measurement and the data processing performed in the worksheet are
displayed.
16
9 The status bar of the document window displays information about the reference and
the measurement (current measured values and measurement time). A progress bar is
displayed during the measurement.
4.2.2 Menu bar and File menu in the document window
Menu bar The menu and function bar of the document window includes functions for file
administration and measurements. The display of the function bar is automatically
adjusted dependent on the current analysis status and the accessories used.
Symbol Description
File menu with function for data administration in the module window
Open the method window and configure the method
Start the individual sample measurement independent of the sequence configured

Measure the sequence specified in the method
The sequence may temporarily change if function called up repeatedly.
Start reference measurement independent of the settings in the sequence
Available for cassette sipper system and APG sampler:

Switch pump on or off and convey sample or stop conveying
Available for APG sampler:

Lower or raise sample cannula
Available for APG sampler with stirring function:
Switch stirrer on or off
Available for Peltier-tempered accessories:

Configure starting temperature in accessories
Stop sequence
The ongoing sample measurement is completed and the sample sequence still
pending is shown. The sample sequence may be re-edited.
Only for kinetic measurements: Pause
Interrupt the current measurement and continue at a later time
Stop
Stop the current measurement without continuing later
File menu The functions in the File menu of the document window are always applied to the
current document. Some functions may overwrite already existing data. In this case,
there will be a security prompt before overwriting the data.
The following functions are available in the File menu of the document window:
Description
File 
New Empty the current document and prepare for a new
analysis
Open method Open a method from a saved document

The method can be modified for subsequent
measurements.
17
Open results Open measured values, method and analyses

Additional data can be added to the document through
measurements with an unchanged method.
Copy / Paste Available in the Spectrum and Colorimetry modules:
Copy measured results and insert into a different document
Sign document Sign a document with an electronic signature
Display / Hide Signaturese Show signatures of a document / close display
Display / Hide Audit Trail Show or hide the audit trail
Save Save measured values, method and analyses in a file
Save as Save measured values, method and evaluations in a new

file
Export measured values to Export measured values of the document window to the
CSV file CSV format
Export Export measured values and analyses via the report

functions
Print Print measured values, method and analyses in accordance
with the selected report template
Creates print template Create your own print templates
Edit document title Change document title in the tab of the document
Open Excel, export measured Export measured values and results to an Excel file and
values and results open in this Excel file
Note: Requires installation of Excel
Close Close the document
4.2.3 Worksheets in the document
The measured values and analyses are displayed in the worksheets of the document.
The Measurement worksheet includes the original measured values which may only be
analyzed with the functions available in the module method. The Measurement
worksheet cannot be deleted, which means the original measurement data is always
retained.
If you wish to edit the measured values using the mathematical functions available in
the module, you will need to open another worksheet with a copy of the original
measurement data in the document.
Function bar in worksheet The worksheet has its own function bar on the top right side:
Function Description
Evaluation Menu with module-specific analyses
+New worksheet Only available in the Measurement worksheet:

Copy measured values to a new worksheet
Data handling Not available in the Measurement worksheet:

Modify data through mathematical operations
Options Scale graph view, create text boxes, rename worksheet
18
Divide the worksheet The worksheet is divided into a bottom and top area. Generally, the measured values are
displayed in the top area and the results of the data analysis in the bottom area. The
boundaries between the areas can be moved freely.
Execution
 Move the mouse pointer on to a boundary line and once the double arrow appears
move the boundary line with the left mouse button held down.
 Move the boundaries into predefined areas by clicking on the separating line arrows
at the ends of the separating lines.
Example of separating line arrows to adjust the window areas
Tip
You will also find the separating line arrows in the method window on the Sample
sequence page between the graphical input and the sample table.
Add new worksheet Execution

 In the Measurement worksheet, click on [+New worksheet].
 In the document, an additional worksheet with the title New worksheet and a
copy of the original data appears. The function bar of the new worksheet
contains the Data handling menu for mathematical data processing.
Tip
Only the measurement data activated on the worksheet will be copied. Deactivate the
measurement data on the Measurement worksheet which does not need to be
processed further (right-click on the sample and select the function Deactivate in the
context menu).
Rename a worksheet A new worksheet can be given a different title. The Measurement worksheet cannot be
renamed.
Execution
 On the worksheet, select Options  Rename worksheet .
 Edit the name in the input field.
 The current worksheet is renamed.
4.2.4 Show spectra and measurement curves on the worksheet
This chapter applies for the following modules:

 Spectrum
19
 Kinetics
 Thermometry
 Color measurement
Spectra or other measured curves are represented in the same way in all modules. The
following functions can be applied to the representation of the curves:
 Scaling curves
 Adding text to curves (text box)
 Marking curves/samples
 Changing the color of curves
 Hiding a curve (deactivating a sample)
Scale curves In the graphical representation of the curves, you can change the display ratio and
enlarge the display. You define the curve section interactively with the mouse or digitally
in an input field.
Run by drawing a frame around the area with the mouse
 With the mouse button held down draw a rectangle across the area to be enlarged.
 When releasing the mouse button the display is updated for the selected area.
 Double-clicking on the curve restores the original display.
Run via dialog window
 Select the Options  Scaling on the worksheet.
 In the list Scaling, select the option User defined.
 In the input fields, enter the X-Axis and Y-Axis of the new limits.
If the Fixed option has been activated, the diagram can no longer be changed with
the mouse.
 The curve display is updated.
Note:
Selecting the option Automatic in the list Scaling will restore the original view.
Create text box A text box is a text field that can be positioned freely on the diagram and be modified in
size. A reference line from the text box can refer to a specific location in the diagram.
This allows for the addition of notes on details of the measurement graph to the
diagram. Text boxes are saved and printed with the data.
Text boxes in a graphic
20
Execution
 Select Options  Create textbox on the worksheet.
The cursor changes to a crosshair with the label "T".
 Click on the position in the curve where the reference line should start and move the
cursor with the mouse button held down to the position where the text box should
be placed.
 Click in the text box and edit the text.
 The text box can be printed and saved.
Additional text box functions
 The size of the text box can be adjusted later. To this end, move the cursor on to one
of the frame lines and move the frame line with the mouse button held down to the
desired position.
 To move a text box, double click on the text box and move the text box to the new
position with the mouse button held down.
 To delete the text box move the cursor on to the upper right corner in the text box.
When the icon is shown, click on it.
Highlight samples For a better overview in a group of graphs graphs can be highlighted using a bold line.
Execution
 Right-click on a sample in the “Measured values” sample table.
 In the context menu, select the item Highlight multiple samples.
 In the selection window, only enable the checkboxes whose samples are to be
highlighted in the graphic and the sample table.
 The selected sample curves are highlighted in bold in the graphic and
highlighted in bold font in the sample table.
Note
Alternatively, individual samples can also be highlighted or their highlighting removed
directly. To do this, select Highlight in chart or Remove highlight in chart in the
context menu.
21
Worksheet with highlighted spectrum
Changing the color of The curves are automatically assigned a color. The colors of individual sample curves
curves may be changed, however.
Execution
 In the Measurement sample table, click on the Color table cell of the sample.
 In the Color dialog, select a new color by mouse click.
 The display of the curve is updated with the new color.
Hide curve / (De)activate Samples can be disabled and hidden from the display. At the same time the sample is
samples excluded from analyses if disabled but not completely deleted from the record.
Execution
 In the Measurement sample table, right-click on a sample.
 In the context menu, select the item Activate multiple samples.
shown and analyzed.
 The disabled sample spectra are deleted from the graphic and highlighted in
light gray font in the sample table.
Alternatively, an individual sample can also be enabled or disabled. To this end, select
the item Deactivate or Activate in the context menu.
4.3 Managing, printing and exporting documents
Documents in ASpect UV have a uniform data structure and include the following
information:
 Measured values
 Method parameters
22
 Analyses of the measured values/calibration graphs

 Parameters for data processing
 Audit trail
The files are identified with the file extension “*.auv” in the Microsoft Windows file
system.
4.3.1 Opening and saving documents
Documents can be opened both in the main window and in the modules. The easiest
way to load measuring results or extract methods is by clicking on the corresponding
icon in the launch bar of the main window. Notes on the configuration of the launch bar
can be found in section "Configuring the launch bar" p. 152.
Open results Using the function Open results, the entire document is loaded to the workspace of
ASpect UV. You can view previous measuring results and analyses, add additional
measured values or edit or amend the analysis.
Note:
You can also open measuring results with the extension “*.dat”, which have been
created with WinASPECT or WinASPECT PLUS.
Execution
 In the launch bar, click on the icon Open results or select File  Open results in the
main window.
 In the Open default window, select the file types “*.auv” for ASpect UV files or
“*.dat” for WinASPECT or WinASPECT PLUS files.
 Select the file and open with [OK].
 The results are displayed in the respective module.
When loading the results, the accessory settings and the connected SPECORD PLUS with
accessories are checked (switched on and initialized or simulated). Additional
measurements may only be added to the document if the accessories currently
connected in SPECORD PLUS are consistent with the accessory settings in the method.
Further analysis of the measurement data is always possible.
Alternatively, you may select File  Open results in the document window of a module.
Note:
The following restrictions for loading into the document window of a module should be
noted:
 Only results created with this module can be opened. Otherwise an error message is
displayed.
 The data are opened in the current document. Previous content of the document is
overwritten after a query. If the underlying file of the current document has already
been saved, there will be no change to its memory content.
 WinASPECT or WinASPECT PLUS files may only be opened in the main window.
23
Open method Using the function Open method, the method is extracted from an existing document.
The method contains the measurement parameters, parameters for analyses and the
parameters of the measuring sequence. The method may be edited or use it to
immediately start a measurement.
When loading the method, it is checked whether the initialized device (accessories and
SPECORD type) are consistent with the settings in the method. If this is not the case, the
method will not be loaded. Install the corresponding accessories or load a suitable
method. If you are working in simulation mode, select the SPECORD type and the
accessories from the settings in the Options window.
Execution
 In the launch bar, click on the icon Open method or select File  Open method in
the main window.
 Select the file from the Open default window.

 The method is loaded and can be edited in the module window.
Alternatively, you may select File  Open method in the document window.
Note:
When loading the method, the same restrictions apply as those for loading results (see
above).
Save document The data are saved in the document window with the standard functions Save and Save
as. Saving the filese, both the original data and any changes are saved in the document.
Execution
 In the document window, select the menu item File  Save if changes are to be
saved to an existing file at the same file location.
 In the document window, select the menu item File  Save as if changes are to be
saved in a file at any file location and optionally also a changed file name.In the
document window, select.
 In the Save as default window, enter or select a file name.
 The files are saved with the selected name.
4.3.2 Creating a new document
A new document is the starting point for creating a method with subsequent
measurement. When starting a module, a new document appears without data.
Alternatively, the current document in the module can also be overwritten by a new
document.
Tip
Several documents may be open at the same time in a module. The documents are
arranged behind each other on the workspace and may be selected via the document
title tab.
24
Execution
 By clicking on a module icon in the launch bar or selecting the module menu item, a
new document is created in the module.
For example: Module  Spectrum or Spectrum.

 A new document with the title New document appears.
 By repeatedly clicking on the module symbol, additional new documents will be
opened in the module.
 If there are various modules open at the same time, click on the module icon to first
return the module to the foreground before further documents are created.
 Alternatively, you may select File  New in the current document. Data already
existing in the current document window will be deleted after a query and the
document updated.
4.3.3 Editing document titles
The document title is displayed in the document tab. If several documents in a module
are open, the document can be identified by means of the title. When creating a
document, it displays the title "New document". The title is first defined in the method
settings on the General page. After completion of the method, i.e. after starting the first
measurements, the document title can be changed via a menu command.
Execution
 Select File  Change document title.
 Edit the name in the New title field.
 The document title is updated.
4.3.4 Tracking changes in the audit trail
All operations in a document are always recorded in an audit trail. The audit trail thus
represents the logbook for the created method, recorded measured values and further
data processing.
Display the audit trail Execution

 To display the audit trail in the module, select File  Show Audit Trail.
 The audit trail is shown in a spreadsheet in the lower half of the worksheet.
 To hide the audit trail, select File  Hide Audit Trail.
25
4.3.5 Print documents
Print document / Send Documents are printed via the LLPrint Preview window in the corresponding modules.
email
LLPrintPreview window with print preview and selection for printing and emailing of report
The LL Print Preview window includes the following print options:
Option Description
Print all pages Print the entire report
Print current page Only print the currently selected page
Save as Export the report in various formats (→ "Export data" p. 29).
Send to Using the email client on the computer, the send the report as
email. The same formats as for export are available.
Execution
 In the document window, select File  Print.
 In the window Open in the folder \\Users\Public\Documents\Analytik Jena\ASpect
UV\Reports, select the report template required for the current module (see below).
 In the LLPrint Preview window, select one of the following options:
 Only when printing the report:
Configure printer if necessary.
 The report is printed or the email is sent.
26
Report templates for print Report templates with varying content are used for printing and exporting the
and export measurement data and analysis results. The selection of a report template can thus
determine the scope and content of the output. There are special report templates
available for every module which are adapted to the measurement data and the possible
analyses.
A set of report templates is installed as standard. If required these templates can be
adapted individually with the report designer "Report/Print module List & Label" by
programmers or experienced users. A description of the Report designer is included on
the installation CD of ASpect UV.
After program installation, the templates are stored in the following folders:
 In the installation path C:\Program Files (x86)\ ASpect UV\Reports
 In the library path \\Users\Public\Documents\Analytik Jena\ASpect UV\Reports
Dependent on the language selected during installation the German or English
templates are installed.
Module Spectrum Template (DE/EN) Content of the printout or export
Spektrum_Komplett.lst/ All available data: method, spectra, calibration,

Spectrum_Complete.lst results table, analyses
Spektrum_Methode.lst/ Method of the document

Spectrum_Method.lst
Spektrum_Audit Trail.lst/ Audit trail of the document
Spectrum_Audit Trail.lst
Spektrum_Messwerte.lst/ Measured values of the document
Spectrum_Raw data.lst
Spektrum_Ergebnisse.lst/ Result table with variavles and statistics and

Spectrum_Results.lst calibration curve (if applicable)
Spektrum_Kalibration.lst/ Calibration curve and calibration parameters
Spectrum_Calibration.lst
Spektrum_Auswertung.lst Analyses of the measured values
Spectrum_Evaluation.lst
Energie_Service.lst For service documents only

Inspection of the UV lamp
Spektrum_Grafik.lst Spectra (with thick lines), results and calibration
Spectrum_Chart.lst curve
Spektrum_Messwerte_Transponiert.lst Transposed output of the measured values of the
Spectrum_RawData_Transposed.lst document
Module Photometry Template (DE/EN) Content of the printout or export
Photometrie_Komplett.lst/ All available data: method, spectra, calibration,

Photometry_complete.lst results table, analyses
Photometrie_Methode.lst/ Method of the document
Photometry_Method.lst
Photometrie_Audit Trail.lst/ Audit trail of the document

Photometry_Audit Trail.lst
27
Photometrie_Messwerte.lst/ Measured values of the document

Photometry_Raw data.lst
Photometrie_Ergebnisse.lst/ Results table (incl. Variables and statistics, if

Photometry_Results.lst activated) and the calibration curve (if available)
Photometrie_Kalibration.lst/ Calibration curve and calibration parameters
Photometry_Calibration.lst
Photometrie_Auswertung.lst Analyses of the measured values
Photometry_Evaluation.lst
Photometrie_Messwerte_Transponiert.lst Transposed output of the measured values of the

Photometry_RawData_Transposed.lst document
Kinetics module Template (DE/EN) Content of the printout or export
Kinetik_Komplett.lst All available data: method, spectra, calibration, results

Kinetic_complete.lst table, analyses
Kinetik_Methode.lst Method of the document

Kinetic_Method.lst
Kinetik_Audit Trail.lst Audit trail of the document.
Kinetic_Audit Trail.lst
Kinetik_Messwerte.lst Measured values of the document
Kinetic_Raw data.lst
Kinetik_Ergebnisse.lst Results table with variables and statistics and

Kinetic_Results.lst calibration (if available)
Kinetik_Kalibration.lst Calibration curve and calibration parameters
Kinetic_Calibration.lst
Kinetik_Auswertung.lst Analyses of the measured values
Kinetic_Evaluation.lst
Kinetik_Grafik.lst Results table, kinetic curves (with thick lines) and

Kinetic_Chart.lst calibration (if available)
Kine- Transposed output of the measured values of the
tik_Messwerte_Transponiert.lst document
Ki-netics_RawData_Transposed.lst
Thermometry module Template (DE/EN) Content of the printout or export
Thermometrie_Komplett.lst All available data: method, temperature curves, results table,

Thermometry_complete.lst analyses
Colorimetry module Template (DE/EN) Content of the printout or export
Farbmetrik_Komplett.lst All available data: method, spectra, results table, color

Colorimetry_complete.lst coordinates/color numbers
28
Validation AJ module, Template (DE/EN) Content of the printout or export

Standard Ph. Eur.,
Compact.lst Short log with overview of validation results
Maintenance economic
Detailed.lst Detailed validation protocol with target values, measurement data or
spectra, validation results and audit trail.
USP module Template (DE/EN) Content of the printout or export
USP_komplett.lst Detailed validation protocol with target values, measurement data or

USP_complete.lst spectra, validation results and audit trail
User management Template (DE/EN) Content of the printout or export
UM_Audit_Trail.lst Audit trail about the changes in the user profiles
UM_Login_List.lst List of successful and failed user logins in ASpect UV
UM_UserList.lst List of user profiles created and their status
UM_UserRights.lst List of rights for a selected user profile
4.3.6 Export data
ASpect UV files with the extension "*.auv" are encrypted and thus protected against
manipulation. However, to allow the use of the data in other programs, the data can be
exported. For the output of data ASpect UV has a large number of output formats
available. Along with being output to the printer, the data can be exported into PDF,
HTML, CSV, Excel or Text format or can be saved as an image file. If there is an Excel
program installed on your computer, you may use the menu command Open Excel.
export measured values and results to export to a new Excel folder.
Export data using the The report functions permit the following export formats:
report functions
 HTML
 PDF
 Rich Text Format (RTF)
 Text format (*.txt)
Tip
Use the text format (*.txt) for output in a format that can be read in a spreadsheet
program or export the measurement data to a CSV file.
The content of the exported file is controlled by the report templates. Various templates
are available for each module:
Execution
 Select File  Export in the document window.
 In the window Open, select the template in the folder
\\Users\Public\Documents\Analytik Jena\ASpect UV\Reports (→ "Report templates for
print and export" p. 27).
 In the window Output settings in the list Output task, select the desired export
format and click on [Start].
29
 In the Save as window enter the name for the export file.
 The document data are exported to the selected format.
Tip
Exporting via Reports is also possible via the Print function. To do this, use the Save as
function in the LLPrint Preview window.
Export measured values in The measured values may be exported to a CSV file.
CSV-format
Execution
 In the document window, select File  Export measured values in CSV-format.
 In the Save as default window, enter or select a file name.
 The data is exported to a CSV file with the selected name.
Export measured values If you have installed an Excel program on your computer, the measured values can be
to Excel directly exported to the program without being saved.
Execution
 In the document window, select File  Open Excel, export measured values and
results.
 Excel opens and the data is exported to a new Excel folder.
4.3.7 Automatically archiving documents
Data can be automatically archived. The following options are available for this:
 Automatic save
 Automatic print with selected report template
 Automatic export with selected report template
 Automatic export to a CSV file
For automatic archiving, the target files and the report template must be specified in the
method. The time and activation of this function is defined program-wide in the
Options window (→ "Activating automatic archiving" p. 159).
30
Defining the target files

and the report template in
the method
Archive automatically page in a photometry method
Procedure
 Switch to the Archive automatically page in the method parameters.
 Activate the archiving function.
 For the automatic save or the automatic export, select the target file. Click on […]
next to Target file and enter the path and the file name in the Save as window.
 Optionally, open a window with options for the target file name with [Advanced]
and define the following:
− Addition in the file name: Add a character string in front of the selected name
(Prefix) or after the name (Suffix).
This function can be used together with the Allow user change before saving
option to add a serial ID in front of or behind the defined sample name, for
example.
− Timestamp: Add the measurement start time to the name.
The start time is in the format yyyy-mm-dd_hh_mm-ss.sss, for example
"autosave_Photometry 2020-10-22_13_59-50.022.auv". The timestamp can
also be added in front of or behind the file name.
This option is activated by default.
− Allow user change before saving: When activated, a window appears with an
entry field to change the file name at the time of archiving. The file path cannot
be changed.
 Select the report template for the automatic print or the automatic export. Click on
[…] next to Report template and select the report template in the Open window.
Note: Automatic archiving must be activated program-wide in the Options / Advanced
/ Automatic archiving window (→ "Activating automatic archiving" p. 159). If archiving
is not activated, a notification will be displayed when a method with automatic archiving
is closed or loaded. The Options window can be opened via the link in the notification
and archiving activated. If archiving is kept deactivated, measurements can be started
with the method, but the data must be saved, printed or exported manually.
31
4.4 Sample sequences
A sample sequence contains a list with the sequence of the samples to be processed,
with information about sample type, name and other data important for the analysis.
After starting the measurement, the sequence is automatically processed and the results
are displayed in the results window. Sample sequences may be viewed during the
measurement in order to check editing status. Further samples may also be added to the
sequences.
4.4.1 Structure of sample sequence and sample types
The sample sequence window is divided into a graphic area on the left and an input
table on the right. In the graphic area, the sample types can be assigned to the samples
by mouse click. In the input table, additional sample properties are specified.
Tip
For the input in the graphic area, the figures in the sample icons should be visible.
Enlarge the method window and move the separating line between graphic and sample
table until the figures are visible.
Separating line arrows between the graphical and spreadsheet area
32
Sample sequence in the method of the Spectrum module
Structure of the sample The Sample sequence window contains the following options and buttons:
sequence window
Option/ Description
button
Mode of Activate a sample type using the buttons and then assign the samples to the
sample type graphic or table per mouse click
[Add Add one or several samples of a sample type to the sequence from a selected
samples] list place in the list
[Remove Remove one or several samples of a sample type from the sequence
samples]
[Import Import sample data from a CSV or TXT file (→ "Supplement C – Data
data] format for importing sample information" S. 212)
Sample table Sample table with specific properties (name, type, note, formula variables (A,
B, …)).
Only for analyses with calibration:
The concentrations of the standard samples are additionally entered. The
dilution and weight of standards and samples are only taken into account in
the result calculation for a concentration analysis with calibration.
33
Sample types ASpect UV uses the following sample types:
Sample type Button Description
reference [R] Reference measurement

Every absorbance, transmission or reflectance measurement
requires a preceding reference measurement. This reference
measurement is usually taken with a reference sample containing
the sample matrix without analyte. The following sample
measurements are corrected with the measured value of the
sample matrix and the baseline (device-specific value).
Note:
This value is saved internally and will not be output.
Sample [S] Sample with unknown analyte concentration
Standard [Std] Sample with known analyte concentration
blank [0] This sample type is only required for displaying the absorbance of
the sample matrix. For this purpose, set the sample sequence to
Reference – Blank value – Sample. The reference measurement is
taken with an empty measuring space (air) or a sample without
reference matrix (e.g. cell with water). The subsequent blank
measurement is taken with the sample matrix and without
analyte. The absorbance caused by the matrix is output as the
blank value.
The following sample measurements are corrected with the
reference value and the blank value.
No [?] For cell changer and APG:
measurement The corresponding measuring space in the accessory is not filled.
No measurement is carried out at this measuring space.
wash [W] For cassette sipper system and APG:
Sample only used for purging the sample paths. No measurement
takes place.
4.4.2 Create sample sequence
Configure sample table The columns of the sample table may be shown or hidden as required:
 After right click on the table header enable the required columns in the context
menu.
Attention At the start of the sample frequency, there should be a reference. If no reference has
been defined in the sequence, a reference measurement must be taken manually before
starting the sequence.
Sample sequences for When using cell holders and accessories for solid samples, the sample must be manually
manual sample changes replaced after every measurement. The samples are consecutively added to the sample
table, without any gaps.
 Click on [Add samples].
 In the window Add samples, enter the following data:
34
Option Description
Paste Selection of insertion area of certain type of sample:

At the start Add samples at the start of the sample table (line 1)
At the end Add samples at the end of the sample table
From line number Add samples in consecutive order according to start of
line number selected
Number Number of samples to be added
Sample Selection of sample type (→ "Structure of sample sequence" p. 32)

type
 Confirm the selection with [OK].

 The samples appear in the sample table.
The sample types may be subsequently edited (→ "Edit sample sequences" p. 37).
Sample sequences for When using cell changers oder samplers, the sample types may also be assigned in
automatic sample batches, whereby one batch corresponds to one accessories load. Not all measuring
changes positions need to be populated without gaps within a batch.
Tip
If not already shown, when using a cell changer or sampler, show the Charge column for
a better overview of the sample placement in the accessories.
 Click on [Proben hinzufügen].

 In the window Add samples, enter the following data:
Option Description
Add Selection of insertion area of certain type of sample:

At start Add samples at the start of the sample table (1st
batch, line 1)
At the end Add samples at the end of the sample table
From line number Add samples in consecutive order according to start of
line number selected
From position in batch Add samples from the selected position in the batch
From first free position in batch Automatic search for the next free
position to add the samples
Sample type Selection of sample type (→ "Structure of sample sequence" p. 32)
Number Total number of samples to be added
Total per Total number of samples within one loading of the accessories regardless of
batch the sample type
 Confirm the selection with [OK].

 The samples appear in the sample table. If the number of samples per batch
differs from the maximum accessories load, the samples are automatically
divided up into the following batches.
35
Example of sample Using various parameter settings, samples of different types may be allocated across
distribution within several various batches in the cell changer or autosampler. In the example, the settings for a
batches: measurement of 4 batches with 5 samples respectively and the associated reference are
applied for the 8-cell changer.
 Click on [Add samples] and first allocate the 4 reference samples to the 1st sample
position of each individual batch of the cell changer:
 Click again on [Add samples] and allocate the samples to the various batches (5
samples * 4 batches = 20; 1 reference + 5 samples = 6 samples in total per batch):
 There are 4 batches created, in each of which there is first a reference, followed
by 5 samples. Per batch (load), there are 2 sample positions that remain empty
in the cell changer.
36
Example of sample sequence for cell changer
Remove samples from the Sample positions that are not required may be removed from the sample table:
sequence
 Click on [Remove samples].
 The following options are available to choose from:
Option Description
Delete At the beginning Delete number of samples at start of sample table

At the end Delete number of samples at end of sample table
Number from line number Delete number of samples from selected line
number
Line number from - to Delete samples between the line numbers specified
Sample all Delete samples, regardless of type

type sample type Samples of selected sample type
Number Number of samples to be removed
4.4.3 Edit sample sequences
The sample sequences may be edited:

 Assign a name, notes, concentration values (for standard samples) and different
variables to a sample
 Change the sample type
 Remove samples from the sequence
Enter sample name  In the sample cell, click on the column Name and enter the name.
Note:
Right-click in the context menu to choose from standard editing functions (cut, copy,
add and delete).
37
Consecutive samples can be assigned the same name or, for numbers with numbering,
also an ascending index:
Highlight the cell with the name and the following cells with the mouse button held down.
Right-click on the marking and in the context menu Fill down with same value Fill or
Down ascending values.
Enter additional sample  When activating the calibration, the concentration of the standard samples must be
data entered in the Concentration column of the sample table
 If applicable, enter the factors for sample weight and dilution for samples and
standard samples.
 In other table columns, individual variables A – H, Note and other sample data may
be entered for each sample.
Tip
Sample weight and dilution are only available for analyses with calibration. For all other
analyses the entries in these columns are ignored. If you want the sample weight and
dilution to be taken into account in a concentration analysis with factor, use the
variables A - H in the formula.
Edit sample types The sample types in the sample table may be edited with the sample switches in the
graphic.
 Enlarge the graphical area by moving the separating line arrows until the sample
number appears on the sample icon.
 Activate the sample type button (Mode of sample type) and click on the sample
icon. Several adjacent samples may be highlighted together by drawing a frame
around the sample icons with the mouse button held down.
 Alternatively, you can click in the sample type field in the sample table or move over
several consecutive samples with the mouse button held down.
 Right-click on the Typ table cell of a sample to open a context menu in which the
sample type may be selected for the sample.
Editing the sample An ongoing measurement may be interrupted in order to edit samples of the sequence
sequence during the not yet measured or to add further samples to the sequence.
measurement
 In the document window, interrupt the measurement with .
 The window Sequence settings appears.
 Add further samples from the sequence or edit any samples not yet edited (see
above) and confirm with [OK].
 Confirm the query "Start measurement?" with [Yes].
 The measurement is resumed.
4.5 Using formulas
In ASpect UV measuring results can be linked in a formula and their results be further
analyzed. In the method window, this function can be found on the Analysis page. The
38
result of a formula calculation can also be used in a calibration. In this case, select the
formula on the Calibration page.
The possible formula arguments (x value) are adapted to the respective module. In the
Photometry module, for example, you will find the measuring results of the selected
wavelengths or in the Spectrum module, automatically searched for peaks. The input of
a formula is the same in all modules except for the selection of the values.
Structure of the formula
editor
Example of formula editor from the Spektrum module
The list below includes the components of the formula editor. The numbers correspond
to those in the figure above.
No. Element Description
(1) Input area The formula can either be compiled using the input elements of the
editor or be entered directly via the keyboard.
The buttons are used to check and correct the formulas:
[Test] Tests the correctness of the formula syntax.
[Back] Deletes the last entry in the formula.
[Clear] Deletes the entire formula.
(2) Number On the number block the numerical entries are made.
block
(3) Block for In the formula, up to 8 (A – H) sample-dependent variables can be

sample used. The value of the variables is entered individually already in the
variables sample table of the sequence or for subsequent calculations after the
measurement in the results table.
(4) Functional The functional area provides the mathematical operators for the
area formula.
(5) Value area Depending on the module selected, various arguments can be linked in
the formula, e.g. defined wavelengths in the Photometry module, peaks
in the Spectrum module or slope in the Kinetics module. The values
selected are transferred to the formula with the icon.
(6) Unit of the The entered unit is output on the worksheet during the analysis.
results
39
4.6 Using accessories in SPECORD PLUS
Connected accessories are detected automatically when switching on the SPECORD

PLUS and are taken into account in the method parameters. This chapter provides an
overview of the accessory-specific method parameters and program functions organized
by accessory type and module.
Please also pay attention to the notes on assembly, adjustment and function of
accessories in the operating instruction "Accessories SPECORD PLUS".
In the method, the accessory parameters are always arranged on the Accessories tab.
4.6.1 Passive accessories
Passive accessories are accessories that are not electronically controlled or automatically
detected via an identification plug:
 Standard cell holder  Thermostated cell holder 10 mm and
50 mm
 Holder for 100mm cells
 Holder for cylindrical cells up to 50
 Holder for round cells
mm and 100 mm
 Holder for microcells
 Optic fiber coupling kit
 Adjustable cell holder for microcells
 Holder for solid samples
Method parameters The following method parameters are available in all modules:
Option Description
Select passive Selection of the accessories listed above

accessory The accessories are displayed in the document. The selection does not
affect the method parameters further.
Change meas. Swap beam paths in the sample space. The beam path for the sample
channels measurement (M) becomes the reference beam path (R) and vice versa.
4.6.2 Cassette sipper system
Option Description
Pump time The pump time required to transport the sample optimally into the cell.
Rinse time Time for the pick-up of rinse samples to rinse the cell
Note:
The order of the samples and rinse samples is defined in the sequence (→
"Sample sequences" p. 32).
Slow run The pump speed has been reduced by up to one half. Thus allowing for the
reduction of degassing or segregation. Smaller sample volumes may be
dosed more accurately.
40
Functions in the When using the cassette sipper system, the following buttons are additionally displayed
document window in the icon bar of the document window:
Button Description
Switch pump on and off
Run With these functions can be used for purgig, draining or filling after
sipper/ measurement.
Stop sipper
Functions in the main The optimum pump time can be calculated with program support using the menu
window command Instrument  Accessories  Optimize pump time (→ "Optimizing the
pumping time of the cassette sipper system" p. 146).
4.6.3 Sampler
The following samplers are available for the SPECORD PLUS:

 APG 49
 APG 64
 APG 116
 APG S
Option Description
Pump time Time for transporting the sample optimal into the cell
Wash time Time for aspiration of the rinsing liquid from the defined rinse sample
spaces for rinsing the cell
Manual Start Start the measurement at a sample only after pressing another button.
This option must be enabled if multiple measurements are run per sample.
Stirrer enable Enable the stirring function for samplers with stirrer
Stirrer speed Stirring speed in percent

The maximum stirring speed corresponds to 100 %.
Stirring time Stirring time for the sample before the sample is aspirated and transported
to the cell.
Test stirring Switch stirrer on and off to determine the optimum stirring speed
Slow run The pump speed has been reduced by up to one half. Thus allowing for the
reduction of degassing or segregation. Smaller sample volumes may be
dosed more accurately.
41
Functions in the When using a sampler the following buttons are additionally displayed in the icon bar of
document window the module.
Button Description
Switch pump on and off
Run/Stop This function can be used to purge, rinse or fill the system additionally or
sipper after the end of the measurement.
Lower or raise the sampler needle in the sample container

Needle
down/up
Switch the magnetic stirrer on and off

Run/Stop
stirrer
Functions in the main In the menu item Instrument  Accessory of the main window, additional functions are
window released for adjusting the sampler (→ "Adjusting the sampler, moving to individual
positions and optimizing the pumping time" See also 148).
4.6.4 Cell changer
The following cell changers available in the SPECORD PLUS:

 Cell carousel
 6-cell changer (also Peltier tempered)
 8-cell changer (also Peltier tempered)
 Simultaneous use of two 6-cell or 8-cell changers in the measurement and reference
channel of the SPECORD PLUS.
 Combination of a Peltier-tempered cell changer with the Peltier-tempered cell holder
The function of the Peltier tempering for the 6-cell and 8-cell changer is explained in
the section "Peltier-tempered accessories" on page 43.
Parameter Description
Cyclic for Only for enabled multiple measurement

Sequence of sample processing
Sample All repeat measurements are first performed at one
sample. Then the next sample in the cell changer is
indexed and the measurements are performed there.
All samples are consecutively processed in this
manner.
Batch First, the first measurement is performed in each
sample in the cell changer, then the cell changer
returns to position 1 and starts the next measurement
in each sample. This is repeated until the last
measurement.
42
Note
If more than one batch has been defined in the sequence, the batches are
processed consecutively.
Manual Every measurement of a sample in the cell changer must be started by pressing
start a button.
Synchronius Only if two cell changers are used:

If enabled, the two cell changers move synchronously. A separate reference
can be used for each sample.
If deactivated, the cell changers are moved consecutively through the beam
paths (offset mode). In this way, both cell changers are used for the sample
measurement. At first, the samples in the reference channel are measured
followed by the samples in the measuring channel. The last cell position of the
cell changer in the reference channel and the first cell position of the cell
changer in the measuring channel are used in offset mode to record the
baseline (device-specific value). They must remain empty or be populated with
identical reference samples.
Functions in the main In the menu item Instrument  Accessory of the main window, the following functions
menu are available for cell changers:
 Index specific individual sample positions
 Switch cell changer on and off
When switching off, the cell changer travels to the park position in facilitate a simple
removal and installation and for packing into the storage box.
 Adjust cell changers
 Detect accessories
 See also section "Adjusting the cell changer and moving to individual positions" p.
145
4.6.5 Peltier-tempered accessories
The following Peltier-tempered accessories are available for the SPECORD PLUS:
 Peltier-tempered cell holder
 6-cell changer, Peltier-tempered
 Two 6-cell changers, Peltier-tempered
 8-cell changer, Peltier-tempered
 Two 8-cell changers, Peltier-tempered
 8-cell or 6-cell changers (in measuring channel) and cell holder, Peltier-tempered (in
reference channel)
The function of the sample positioning with the cell changer has been explained in
section "Cell changer" p. 42.
43
Functions in the The following buttons are additionally displayed in the icon bar of the document
document window window:
Button Description
Set the accessory temperature to the defined value (Fixed temperature

Set mode) or the starting temperature of the temperature curve (Variable
Temperature temperature mode).
With the Peltier-tempered accessories the following temperature operation modes are
possible:
 None
 Fixed temperature
 Variable
Temperature mode none The temperature mode None is available in the modules Photometry, Spectrum,
Colorimetry and Kinetics. In this mode, the temperature of the cell holder/changer is
not controlled via the ASpect UV program, but can be directly specified at the
temperature control unit (see also the operating instructions on "Accessories for
SPECORD PLUS").
After clicking on [Info], the information about the available temperature range and the
actual temperatures appear.
Temperature mode Fixed The temperature mode Constant is available in the modules Photometry, Spectrum,
temperature Colorimetry and Kinetics. In this mode, a constant temperature is configured and kept
for the entire measuring period. The measurement is taken 5 s after reaching the
temperature within the agreed temperature accuracy range plus a potential waiting
period.
Temperature None Set a constant temperature.

mode
Temperature [°C] Enter the target temperature
Precision of Select the accuracy of the temperature control

temperature
Waiting time Enter the waiting time from reaching the target temperature to starting
the optical measurement
Cell sensor active At the start of the measurement, record the registered cell temperature
[Info] Display information about the adjustable temperature range and the
current temperatures in accessories
44
Method configuration for constant temperature mode of the Peltier-tempered accessories
Temperature mode The temperature mode variable allows you to record temperature-dependent
variable measurement curves. Each measurement is taken 5 s after reaching the temperature
within the agreed temperature accuracy range plus a potential waiting period.
The temperature mode variable is available in the modules Thermometry, Colorimetry
and Spectrum. In the Thermometry module, the measurement is carried out at a
wavelength (→ “Thermometry“ p. 108). In the Spectrum module, a spectrum is recorded
for every temperature specified. A section may be applied to the spectral group at a
wavelength and the intersection curve evaluated in the Thermometry module.
Temperature Variable Recording of a temperature curve (Thermometry

mode module) or a spectral group (Spectrum module or
Colorimetry)
Precision of Select the accuracy of the temperature control
temperature
Cell sensor At the start of the measurement when the target temperature is reached,
active the registered cell temperature is recorded. The measured value
(absorbance / %T) is allocated to the cell temperature. The temperature
measurement is thus more accurate with regard to the sample temperature.
If the option is disabled, the block temperature (standard temperature) is
recorded or indicated as an abscissa value in the Thermometry module.
Temp. after If enabled, the set temperature is approached and held after the last optical
measurement measurement.
Temperature Use [setup] to open the Variabler Temperaturbetrieb window to enter the
values temperature values
[Info] Show information about the adjustable temperature range and the current
temperatures in accessories
Method configuration for variable temperature mode of the Peltier-tempered accessories
45
After clicking on [Setup], the window Variable temperature mode appears. The
parameters of the temperature stages can be edited individually in the table rows or
defined automatically using the checkboxes and input fields.
Option Description
[Insert] Enter a new temperature step after the highlighted table row
[Delete] Delete the highlighted table row
Gradient from Automatically generate a temperature gradient from a highlighted table

marked row. Starting with the start temperature of the highlighted table row
position further temperature stages are inserted into the table in steps until the
final temperature has been reached.
Diff. [°C] Temperature difference of a temperature stage
to °C Final temperature of the gradient
[Add] Add temperature gradient at the highlighted
position
Waiting time Waiting time from reaching the target temperature to starting the optical
for all measurement
With [Set], the temperature entered is transferred to all temperature
stages of the table.
Note: Regardless of the set waiting time, the optical measurement is
carried out no less than 5 s after reaching the temperature within the
agreed temperature accuracy range in the block. The waiting time is added
to this 5 s period.
[Undo] Reverse the last action
Optical Yes Perform an optical measurement at temperature

measurement step
No No optical measurement takes place at the
temperature step.
46
Configuration of the temperature stages in the Variable temperature mode window
4.6.6 Integrating sphere
The integrating sphere is used for the measurement of scattering solid or liquid samples
in transmittance, absorbance or diffuse reflectance.
Method parameters In the method parameters, the following options are released under Measuring mode:
 Transmittance
 Absorbance (not with Colorimetry)
 Reflectance
4.6.7 Reflectance measuring attachments
The following reflectance measuring attachments are available for the SPECORD PLUS:
 Absolute reflectance attachment
 Reflection measuring attachment 11° 60°
Method parameters In all modules, only the Reflection option is available as measuring mode.
47
4.6.8 Scanning attachment for solid samples
The scanning attachment for solid samples is used for the locally resolving
determination of absorbance or the transmission of small transparent solid samples.
Option Description
Start Start coordinate of a measurement
Finish End coordinate of a measurement
Step Measuring point spacing on the sample

width
[Set] Move the scanning attachment to the start or end coordinate of the
measurement
[Zeroth Set the monochromator grating to the zero order

order] The undispersed (white) light of the Vis lamp passes through the sample
chamber and due to its good visibility facilitates the adjustment of the sample in
the scanning attachment.
Method parameters for the scanning attachment for solid samples
48
Quick measurement module and online measurements
5 Quick measurement module and online measurements

Quick measurement The Quick measurement module is intended for the measurement of one absorbance or
module transmittance value. The data is only displayed on screen and is neither saved nor
printed. Optionally, the value can be determined during a single measurement or
automatically continuously. In this measurement, electrically controlled accessories are
not supported. The cell changer can only be used as cell holder; the cassette sipper
system or the autosampler cannot be used.
Structure of Quick
measurement window
Quick measurement window for determining absorbance values
Option / button Description

With manual start,
Measure
start measurement.
Set zero Determine reference value. The reference value is used as a

base value, i.e. is set to zero.
Exit the measurement window.
Close
Value display Displays the measured absorbance value.
Waevelength and slider Enter the wavelength in the input field or set it via the slider.
Display List for measured values: Absorbance or Tansmittance
Measure automatically, Determine the measured value continuously every 5 seconds.

interval 5 sec
Manual start
Determine the measured value after clicking on .
Manual start In this mode you select a wavelength and start the measurement via mouse click.
Execution
 Select Module  Quick measurement.
 Edit the wavelength in the input field or set it via the slider.
 Enable the option Manual start.
49
 Insert reference sample into the cell holder where applicable and click on .
The reference measurement is performed. In the display, the value "0.000" is shown.
 insert samples into the cell holder and click on .

 The results display is updated. Read the measured value.
 For subsequent sample measurements insert the next sample into the cell holder
and click on .
Continuous measurement In this mode a measured value is determined continuously at time intervals of 5 s.
Execution
 Select Module  Quick measurement.
 Edit the wavelength in the input field or set it via the slider.
 Enable option Automatic measurement, interval 5 sec.
 Insert reference sample into the cell holder and click on .

The reference measurement is performed. In the display, the value "0.000" is shown.
 Insert samples into the cell holder.
 Wait until the new measured value is displayed and read the measured value.
 For the subsequent measurement, insert samples and wait each time until the
measured value is updated.
Online measurement During the online measurement, the measured values are permanently determined at
the currently set wavelength at one second intervals and output to the launch bar of the
main window (→ "The main window in ASpect UV" p. 12). The online measured values
are only shown after the first measurement. The online measurement may be enabled
in the window Options / Launch bar (menu command Settings  Options).
Like the module Schnellmessung, the online measurement can be used for a quick
sample value determination. The measurement is not saved and cannot be printed. To
use the online measurement, the wavelength must be set in the quick measurement and
the reference value determined (set to zero). The measurand shown complies with the
value last measured in the active document.
Execution
 Activate the window Quick measurement.
 Set the wavelengths and click on [Set zero].
 Close the window Quick measurement.
 Insert samples into the cell holder and read the current measured values in the
launch bar.
 The measured values are continuously updated.
50
Displaying online measurement
51
Photometry module
6 Photometry module
The Photometry module is based on the measurement at selected wavelengths. Based
on this the measured values of the individual wavelengths can be analyzed in a
calibration or formula. The method thus meets the requirements for quantitative routine
analysis.
Open Photometry module Execution

 Open the Photometry module by clicking on the button in the launch bar of the
main window:
Alternatively, select the item Module  Photometry in the menu.

 A new document is opened in the workspace. All functions of the window are
assigned to the Photometry module.
 With each additional click on the button Photometry in the launch bar, another new
document is opened in the workspace.
6.1 Method configurations in Photometry module
This chapter includes all configurations that can be made for a method in the module
Photometry. Please also pay attention to the notes in chapter "What you should know
about ASpect in advance" p. 11
Tip
In the ASpect UV program, various methods for enzymology, water analysis and
biochemical analyses have already been installed (→ "Supplement A - Default methods"
p. 208).
Creating a method Execution
 Open the method by clicking on [Method setup].

 Configure the method on the pages of the window Photometry - Settings.
 End the input with [OK].
 The method parameters are output on the left side of the document window on
the Method tab. The buttons for starting the measurement appear.
6.1.1 Photometry – Settings / General
In the Photometry - Settings / General window, the basic measurement configurations

are made.
52
Photometry module
Photometry – Settings / General window with configuration of the measurement parameters
Title Enter the title of the document. The title is displayed in the tab of the document (→
"The document window" p. 15).
Parameter Select measurement parameter configuration:
Option Description
Measurement Select Transmittance, Absorbance or Reflectance (only available with

mode reflection accessories and integrating sphere) as y-axis display.
Wavelengths List of wavelengths for the analysis
 Click on , enter the desired wavelength in the input field and
transfer it to the list with .
 To remove a wavelength from the list, highlight the wavelength in the
list and click on .
Integration Select the time for the recording a measuring point.
time
Slit For SPECORD 210/250 PLUS

Select spectral width (optical resolution):
0.2; 0.5; 1; 2; 4 nm
* The 0.2 nm slit is only displayed in the SPECORD 210/250 PLUS where it has actually
been installed.
Lamp change Select wavelength of change from UV lamp to Vis lamp

at The preset lamp change at 320 nm guarantees an optimal distribution of
energy across the entire wavelength range of the spectrometer. If you are
only working in UV or Vis range, you may also measure the selected
analytical range with the options UV only or Vis only.
53
Photometry module
Multiple measurement Multiple measurements may be carried out on a sample. For the subsequent calculations
and analyses, the average value of the individual sample measurements will be used.
Option Description
Number Number of repeat measurements

From a value > 1 the multiple measurement is enabled.
Cyclic for Sequence of sample processing
Sample All repeat measurements are first performed on one
sample before the measurements start at the next
sample.
Batch The first measurement is consecutively taken in each
sample of the sequence, then the next measurement
starts in each sample. This continues until all repeat
measurements have been completed.
Note for cell changer or APG
One load of the cell changer/APG is measured at a time. Once all repeat
measurements for this load (either by sample or batch) have been processed,
the samples are replaced in the accessories and the next load is measured.
Time If enabled, the repeat measurement takes place automatically after the interval
controlled time entered.
If disabled, the individual measurements are started manual by pressing a
button.
Interval Enter the time after which the measurement is repeated.

time Note:
If the set interval time in a cell changer/APG during batch mode is shorter than
the time required to measure the entire batch, the shortest possible interval will
automatically be used. The results are mapped to the actual times.
6.1.2 Photometry – Settings / Accessory
The parameters in the window Photometry - Settings / Accessory depend on the

installed accessories. The configurations have been described in section "Using
accessories in SPECORD PLUS" p.40.
Photometry – Settings / Accessory window with setting for passive accessory
54
Photometry module
6.1.3 Photometry – Settings / Evaluation
In the Photometry - Settings / Evaluation window, the following settings are made:
 Input of formulas
 Input of a limit value for the measuring results
Photometry – Settings / Evaluation window
Formula If enabled, the measured values are linked in a formula (→ "Using formulas" p. 38). This
function can be used, for example, for a quantitative analysis with factor.
The following functional arguments can be used:
 Measuring result of a wavelength (e.g. "w500")
 Standard deviation of a multiple measurement (e.g. "std500")
Limit value If enabled, an upper or lower limit value is selected for the measured
absorbance/transmittance. Measured values that are above or below the limit value are
marked with "!" in the results table.
6.1.4 Photometry – Settings / Calibration
In the window Photometry – Settings / Calibration, the configuration for a quantitative

analysis with calibration is defined.
Photometry – Settings / Calibration window
55
Photometry module
Calibration setting For concentration determinations with standards, activate the Calibration option:
Option Description
Choice Create new Record calibration within the subsequent measuring

series. The standard samples and their concentrations
must be defined in the sample sequence.
Load from file Load calibration parameters from an existing method.
Regression Select regression. Three types are available:
y=B*x
y=A+B*x
y = A + B * x + C * x2
Ordinate Select the ordinate value for the calibration:
value Use wavelength Select a measured value at one of the wavelengths for
the calibration in the list field. The wavelengths are
determined on the General page.
Formula Use the result of a formula calculation for the
subsequent calibration. After clicking on the
formula editor appears (→"Using formulas" p. 38).
Note:
The formula used on the Calibration page is independent of any formula
entered on the Analysis page.
Unit Enter a unit for the analysis result.
Pathlength Enter the pathlength of the cell.

This value is for information only and is saved with the method parameters.
Limit value If enabled an upper and lower limit value is set for the analysis result. Results
above or below this limit value are identified with "!" in the results table.
6.1.5 Photometry – Settings / Sample sequence
On this page, a sample sequence may be entered (→ "Sample sequences" p. 32), which
may be started with the button in the document window. A sample sequence may
also be created before a measurement.
Requirements for sample  Activating the calibration requires entering a sequence with standard samples.
sequence
 When using cell changers and APG the samples are assigned to the sample spaces in
the accessories.
 If the sample sequence is empty, the Sequence settings window will first appear
when starting the measurement of the sample sequence. This window may only be
exited once there has been at least one sample defined.
6.1.6 Photometry – Settings / Archive automatically
Data can be saved, exported and printed automatically. Select the target files and report
templates in the Archive automatically page (→ "Automatically archiving documents" p.
30). Additionally, automatic archiving must be activated program-wide in the Options
56
Photometry module
window and the times for the individual archiving functions must be selected (→
"Activating automatic archiving" p. 159).
6.1.7 Photometry – Settings / Notice
A note on the measurement may be optionally entered on the Photometry – Settings /

Notice page.
6.2 Performing measurements in Photometry module
The measurement may be carried out if a method has been newly created or loaded in
the document window. During the first sequence measurement start, the sample
sequence included in the method is always processed. Other individual sample
measurements or sample sequences can then be appended.
To start measurements the following buttons are shown:
Button Description
Start the individual sample measurement independent of the sample

[Measure sequence configured
sample]
Start sample sequence
[Measure If you have not saved any sample sequences to the measurement
sequence] parameters, the Sequenz einstellen window will appear after clicking on
this button. This window may only be exited once there has been at least
one sample defined.
Measure a reference
[Reference] A separate reference measurement is required if the sample sequence does
not start with a reference measurement or no reference is available that
matches the current method parameters.
Other accessory-specific buttons have been described in section "Using accessories in

SPECORD PLUS" p.40.
Measure reference If the sample sequence does not have a reference at the first location, the reference
must be recorded separately:
 Place the reference into the sample space.
 Click on [Reference] and confirm the prompt about the reference

measurement.
 The reference measurement is run.
All subsequent measurements are corrected with this reference until the measuring
parameters are modified by a new method configuration or the next reference
measurement takes place.
Start first sequence With the start of the sample sequence the samples are processed in the order specified
in the sample table.
57
Photometry module
Execution
 Place the first sample of the sequence in the sample space or fill cell changer/APG
with samples.
 Click on [Measure sequence].

 Follow the subsequent on-screen prompts for inserting the samples and starting
sample measurements.
 The sample sequence is processed. The measured values and analyses are
displayed in the document window.
Interrupt or stop An ongoing measurement may be interrupted and then continued or stopped
measurements completely. To do this, click on the corresponding button:
Button Description
Stop measurement
Measurement data recorded up to this point remain intact and may be
processed further.
Interrupt measurement and continue at later stage
[Suspend The current measurement step is stopped and then the processing
sequence] interrupted. The measurement pause may be used to view and edit the
ongoing sample sequence (→ "Edit sample sequences" p. 37).
Append next sequence After processing the first sample sequence, the sequence may be started again or re-
edited on other occasions. This is also possible if the document has already been saved
and re-opened to display the results.

The Sequence settings window appears with the sequence stored in the method.
 If necessary, compile a new sequence (→ "Sample sequences" p. 32).
 Confirm the sequence with [OK].
 The current sequence is processed and the data appended to the existing
measurement table/analyses.
Performing single Regardless of the sequence configured, single samples of sample type Probe may be
measurements measured and appended to the measurement table/analyses.
 Place the sample into the sample space.
 Click on [Measure sample].
 Follow further on-screen prompts for starting the measurement.
 The measurement is run and the data are appended to the existing
Tip If measurements of other sample types are to be appended, use the function
[Measure sequence].
58
Photometry module
Re-measuring samples Outliers of a measurement may be re-measured and thus values from a measuring
series that differ greatly replaced by new values. The re-measurement of samples is
always recorded in the audit trail. For the re-measurement of a probe, you will need to
switch to the Measurement worksheet.
 Right click on the row of the sample to be re-measured.
If performing multiple measurements, individual measurements may be re-
measured by right-clicking on the individual measured value.
 In the context menu, select Remeasure sample.
 The measurement is run and the measured value replaced in the sample table.
Existing analyses will be updated.
Attention If additional worksheets besides the worksheet Measurement are already available in
the document, these are not updated during a re-measurement. Empty values may not
be re-measured.
When using cell changers or APG the sample to be re-measured must be placed in the
same position in the accessories. The position in the accessories can be displayed in the
sample table on the worksheet (→ "Displaying, analyzing and editing results in the
Photometry module" p. 59).
6.3 Displaying, analyzing and editing results in the Photometry module
This section explains the particularities of the results display and analysis in the
Photometry module. Please read up on the general structure of the document windows
in section "The document window" p. 15.
59
Photometry module
Photometry document
window
Document window in Photometry module with measured values and quantitative analysis
1 Measurement worksheet 3 Spreadsheets with analyses
2 New worksheet worksheet
Measurement worksheet The Measurement worksheet includes the sample table with the original data. This has
been arranged in the top part of the worksheet.
In the bottom part of the worksheet, data analyses already defined in the method have
been arranged in individual spreadsheets. These analyses can be amended and edited
without changing the original measured values (→ "Analyzing photometry data" p. 62).
Sample table The display of the sample table can be configured freely. After right clicking on the table
header the following parameters can be selected in the context menu:
 Number in the sequence
 Position in the accessories when using cell changer or sampler
 Batch when using cell changer or sampler
 Sample name
 Sample type
 Date/time
 Measured value or average value for multiple measurements
 Standard deviation for multiple measurements
 Temperature for methods with Peltier-tempered accessories
 Notes
 Sample concentration for concentration determination through calibration
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Photometry module
 Dilution and sample weight (only taken into account for concentration determination
through calibration)
 Variables A – H
Via Drag&Drop the order of the table columns can be changed.
For multiple measurements the average values is displayed in the sample table. The
individual measurements are listed adjacent in a separate table.
Sample table with average values from multiple measurements and values from individual measurements
New worksheet The New worksheet worksheet is created after clicking on +New worksheet in the
Measurement worksheet. The original data is first copied to the new worksheet and can
then be edited, e.g. by multiplication with a factor (→ "Editing photometry data" p. 65).
6.3.1 Edit sample table in Photometry module
Edit sample table The following parameters of the sample table can be edited later:
 Sample names
 Notes
 Dilution and sample weight for analysis with calibration
Execution
 Click on the table cell and edit the value.
 The changes are documented in the audit trail. Existing analyses are re-
calculated and updated.
Highlight sample Samples may be highlighted in the sample table. The samples are then shown in bold
font.
Execution
 Right click on a sample in the sample table.
 In the context menu, select the item Highlight multiple samples.
 In the selection window, enable the sample checkboxes.
 The selected samples are shown in bold font in the sample table and
highlighted with a thick outline in the calibration graph display.
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Photometry module
Note
Alternatively, individual samples can also be highlighted or their highlighting removed
directly. To this end, select the item Mark or Remove mark in the context menu.
Sample table with highlighted sample
Enable/disable samples Samples may be disabled in the sample table. The disabled sample is not taken into
account during the analysis but it is not deleted from the record. A disabled sample may
be reactivated.
Execution
 Right click on a sample in the sample table.
 In the context menu, select the item Activate multiple samples.
shown and analyzed.
 The analysis results are re-calculated. The disabled samples are highlighted in
light gray font in the sample table.
Note
Alternatively, an individual sample can also be enabled or disabled. To this end, select
the item Deactivate or Activate in the context menu.
6.3.2 Analyzing photometry data
The data analyses offered in this method can also be performed later. The functions
have been compiled in the menu Analysis that is available on every worksheet. The
analyses always relate to the measured values of the respective worksheet. If the
measured values are edited, the analysis on this worksheet is adjusted accordingly.
The analysis are output in their own spreadsheet in the bottom part of the worksheet.
Edit analysis The analysis parameters can be edited for existing analyses.
Execution
 Click on Settings in the spreadsheet of the analysis and change the input
parameters in the dialog window.
Delete analysis Analyses that are not required may be deleted.

Execution
 At the top right of the spreadsheet of the analyses click on .
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Photometry module
Formula Use the Formula function to link the measured values of a sample in a formula.
Execution
 Select Evaluation  Formula on the worksheet.
 Click on and create the formula with the formula editor (→ "Using formulas" p.
38).
 If required, add further formulas in the same manner.
 The table with the results from the formula calculations is displayed.
Spreadsheet with formula analysis
Tip
Only the measured values of one sample can be used with this function. The Formula
function in the data analysis permits the linking of measured values from different
samples (→ "Editing photometry data - Form" p. 66).
Calibration For measurement series without standards, the Calibration function permits the
calibration data to be loaded from other files and used for a quantitative analysis.
During this time, only the calibration parameters will be loaded. The calibration can
therefore not be combined and re-calculated with standards from the current document.
Execution
 Select Evaluation  Calibration on the worksheet.
 Enable the Calibration option.
 Click on [Open] and select the document with the calibration graph.
 The Calibration spreadsheet with the current calibration graph and the
calculated calibration parameters is displayed.
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Photometry module
Calibration graph display
Spreadsheet with a calibration
The Calibration spreadsheet contains the following information:

 Source of the calibration graph (measured in this document or loaded from a file)
 Regression model
 Analyses wavelengths or calibration formula used
 Regression coefficients A, B and C if calculated in the regression model
 Coefficient of determination
 Pathlength of the cell (for information only, not used in the calculation)
 Date/time of the measurement
Display of the concentration results

The results of the concentration analysis with calibration are entered directly into the
sample table. Concentrations outside of the calibrated range are placed in brackets, e.g.
"(45.3)".
Limit The Limit function permits the identification of measured values above or below a
defined value.
Execution
 Select Evaluation  Limit on the worksheet.
 Enter the value in the input field Limit.
 In the list, select the option Upper limit or Lower limit.
 On the spreadsheet Limit value the samples that are below/above the specified
value are shown. In addition the values are identified in the sample table in
bold and with a "!", e.g. "0.426!".
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Photometry module
Limit value spreadsheet with sample values outside the range
6.3.3 Editing photometry data
Run data editing Data is only ever edited in the copy of the measured values in the New worksheet. The
original measured values in the Measurement worksheet are not modified by these
operations but remain intact. When saving the documents the edited and the original
data are saved.
Attention If the original data in the Measurement worksheet changes, e.g. due to re-
measurement or by disabling a sample, the data in the New worksheet is not updated.
The following functions are available in the Data handling menu:

 Addition of an offset
 Multiplication with a factor
 Convert values to transmission [%T] or absorbance
 Link values from various samples in a formula
The data editing operations are applied to all samples in the sample table.
Execution
 In the Measurement worksheet, click on [+New worksheet].
 In the Data handling menu select the desired function.
Reverse data editing The operations report can be found on the left-hand side of the document on the
Worksheet tab. Here, the operations are listed one after the other. By clicking on the
corresponding operation, the editing step is displayed. Operations can be reversed on
this tab.
Execution
 Right click on the last operation to be reversed.
 Select one of the two options in the context menu:
− Copy to a new worksheet up to this step
A new worksheet is opened and the data is copied with the change history up to
the selected operation. All operations remain intact in the previous worksheet.
− Delete operation
All steps including the selected operation are deleted in the current worksheet.
65
Photometry module
 The operations are reversed in accordance with the selected option.
Data analysis report
Offset With an offset, a constant is added to the measured values. The value of the constant
may be both positive or negative. This allows for interferences to be simulated or
compensated.
Execution
 Select Data handling  Offset on a new worksheet.
 In the field Offset enter the value.
 The measured values and analyses are re-calculated and the displays updated.
Factor With the function Factor the measured values are multiplied by a constant. The
multiplication of an absorbance value with a constant theoretically corresponds to a
change of the pathlength of the sample and/or the concentration.
Execution
 Select Data handling  Factor on a new worksheet.
 In the Factor field enter the value.
 The measured values and analyses are re-calculated and the displays updated.
Change values to This function permits the conversion of the measured values from absorbance to
transmission [%T] / transmission [%T] and vice versa.
absorbance
Execution
 Select Data handling  Convert values to transmission [%T] (Convert values to
absorbance) on a new worksheet.
 The measured values are converted and the display updated.
Formula The Formula function permits the mathematical linking of the measured values of
different samples.
Execution
 Select Data handling  Formula on a new worksheet.
 Enter the mathematical operation in the formula editor (→ "Using formulas" p. 38).
The samples can be selected as arguments in the dropdown list Select sample and
inserted into the formula by clicking on . The samples are numbered in

accordance with their order in the sample sequence and identified by their names.
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Photometry module
 The formula result is calculated and the display updated.
6.4 Example measurement for the Photometry module
In the example, the concentration is determined in 3 samples using the formula

"c = A(446 nm) * 24.3". The standard cell holder is used for the measurement. The gray
filters of the Hellma standard filter set can be used as samples.
The following steps must be carried out:
 Create the document in the module.
 Open method and enter parameter.
 Start measurement.
Preparing the document

 Click on [Photometry] in the launch bar of ASpect UV or select Module 
Photometry.
Define method  Click on [Method setup].

parameters
 Enter the parameters on the pages of the Method window (see below).
 Exit the parameter input by clicking on [OK] in the Photometry - Settings window
and return to the document window.
Parameter entries Enter the parameters as shown in the screenshot.

General page:
Enter 446 nm as wavelength:

 Double click on the preset value "500".
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Photometry module
 Enter the value "446" and transfer it to the list with .

Note: The slit setting only applies for the SPECORD 210/250 Plus.
Accessory page: Make no entries.
Evaluation page:
Enter the formula "w446 * 24.3":
 Click on . The formula editor appears.
 Select the value "w446" from the list and click on .

 Click on "*” and then enter "24.3".
 Enter "mg/l" in the field Einheit der Ergebnisse. Confirm the entries with [OK].
Calibration page: Make no entries.
Sample sequence page:
 Click on [Add samples] and set a reference at the start of the sample table:
68
Photometry module
 Click on [Add samples] once again and add 3 samples to the end of the sample
table:
 Enter "Reference” in the Name field of the reference line and confirm with ENTER.
 In the 2nd line, enter "Sample 1" as the name and then confirm with ENTER.
Highlight the three successive sample name fields, incl. the "Sample 1” field, with
the mouse button held down.
Right-click on the highlighted choice and in the context menu, select Fill with
ascending values.
Note: If the sample icons and type buttons are insufficiently visible in the graphic area,
the separating line between the areas of graphic and sample table can be moved by
clicking on the separating line arrow.
Archive automatically page: Make no entries.
Note page: Enter the text "Example of a concentration analysis with factor".
Running the  Click on in the document window.

measurement
The start information for measuring the reference is displayed.
 Insert the reference sample and click on [Yes] to start the measurement.
The reference measurement is performed. A prompt to measure sample 1 is
displayed.
 Insert sample 1 and click on [Yes] to start the measurement.
 Measure sample 2 and sample 3 in the same manner.
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Photometry module
Example of concentration analysis with factor in Photometry module
70
Spectrum module
7 Spectrum module
In the Spectrum module, spectra or spectral sections are recorded. The Spectrum
module provides the following functions for analyzing and editing spectra:
 Quantitative analysis with calibration function
 Analysis of measured values at defined wavelengths of the spectrum
 Comprehensive mathematical data processing of the spectra
 Automatic peak search
 Apply a section to the spectral group (result of a multiple measurement or a
measurement with Peltier-tempered accessories)
Open Spectrum module Execution

 Open the Spectrum module by clicking on the button in the launch bar of the main
window:
..
Alternatively, select the item Module  Spectrum in the menu.
assigned to the Spectrum module.
With each additional click on the button in the launch bar, another new document is
opened in the workspace.
7.1 Method configuration in the Spectrum module
This chapter includes all configurations that can be made for a method in the Spectrum
module. Please also pay attention to the notes in chapter "
What you should know about ASpect in advance" p. 11.

 Configure the method on the pages of the window Spectrum - Settings.
 End the input with [OK] .
 The method parameters are displayed on the left side of the document window
on the Method tab. The buttons for starting the measurement appear.
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Spectrum module
7.1.1 Spectrum - Settings / General
In the Spectrum - Settings / General window, the basic measurement configurations

are made.
Spectrum - Settings / General window with configuration of the measurement parameters
Title Enter the Title of the document. The title is displayed in the tab of the document (→
Option Description
Measurement Select Transmittance, Absorbance or Reflectance (only available with

mode reflection accessories and integrating sphere) as ordinate display.
Range Enter the wavelength range for the measurement in the input fields.
Measuring Select distance between measuring points.

points Adjustment range: 0.02 to 10 nm
Speed Select measuring speed.
Adjustment range: 0.02 to 200 nm/s
Integration Time for recording a measuring point

time This value is calculated automatically.
0.2; 0.5; 1; 2; 4 nm
been installed.
72
Spectrum module

Multiple measurement For multiple measurements of a sample, the individual spectra are presented in the
results display. For a repeat measurement with time resolution, sections at selected
wavelengths can be inserted in the group of spectra.
Option Description
Number Number of repeat measurements:

From a value > 1 the multiple measurement is enabled.
Cyclic for For cell changer or APG
Select sequence of sample processing.
Sample All repeat measurements are first performed on one
sample before the measurements start at the next
sample.
Batch The first measurement is taken in each sample of the
sequence, then the next measurement starts in each
sample. This continues until all repeat measurements
have been completed.
One batch of the cell changer/APG is measured at a time. Once all repeat
the samples are replaced in the accessories and the next load is measured.
Time If enabled, the repeat measurement takes place automatically after the interval
controlled time entered.
If disabled, the individual measurements are started manual by pressing a
button.
Interval Enter the time after which the measurement is repeated.
time Note: If the set interval time in a cell changer/APG during batch mode is shorter
than the time required to measure the entire batch, the shortest possible
interval will automatically be used. The results are mapped to the actual times.
7.1.2 Spectrum – Settings / Accessory
The parameters in the window Spectrum - Settings / Accessory depend on the installed
accessories. The configurations have been described in section "Using accessories in
SPECORD PLUS" p.40.
7.1.3 Spectrum – Settings / Evaluation
The following analyses may be defined in the window Spectrum – Settings /

Evaluation:
 Peak determination
 Display of the measured values at selected wavelengths
73
Spectrum module
Spectrum – Settings / Evaluation window
Determine peak By enabling the checkbox Determine peaks extreme values are automatically searched
for based on specified criteria. The peaks are highlighted in the spectral display after the
measurement and output to a separate analysis table. Peaks can be used in a formula.
Option Description
List of the complete spectrum Search for all peaks in the measuring range
that meet the criteria below.
only in defined ranges of the spectrum Enter different spectral ranges
in which peaks are searched for.
Type of peak Select the type of peak: Minima, Maxima or Minimum and Maximum.
Show peaks Absolute limit for displaying the peaks

only Peaks are only displayed if they are above or below the ordinate value
entered.
Threshold Height of peak taking baseline into consideration
Difference of peak height between local minimum/maximum and the
subsequent neighboring local maximum/minimum.
Select measured values at Selected measured values are displayed in the sample table and can be used in the
defined wavelengths analysis with formula or in the calibration.
Click on the option Values on defined wavelengths to select the measured values. Then
open an input field with and consecutively enter the desired wavelengths.
Formula If enabled, the measured values are linked in a formula (→ "Using formulas" p. 38).
The values entered above can be used as functional arguments:
 Absorbance/transmission of peaks in selected partial ranges
 Measured values at defined wavelengths
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Spectrum module
7.1.4 Spectrum – Settings / Calibration
On the Spectrum – Settings / Calibration page, you can define an analysis with
calibration. To this end the Calibration checkbox must be enabled.
Spectrum – Settings / Calibration window
Calibration configuration For concentration determinations with standards, activate the Calibration option:
Option Description

series. The standard samples and their concentrations
must be recorded in the sample sequence.
Load from file Load calibration parameters from an existing results
file.
y=B*x
y=A+B*x
y = A + B * x + C * x2
Layer Enter the layer thickness of the cell.

thickness This value is for information only and is saved with the method parameters.
above or below this limit value are identified in the results table with "!".
Ordinate values The Spectrum module can use different values from the spectrum for the calibration:
Ordinate Explanation
value
Value at Measured value at one of the wavelengths

wavelength
75
Spectrum module
Formula Result of a formula calculation

After clicking on the formula editor appears (→"Using formulas" p. 38). As
functional argument, the peaks in defined partial ranges or the values at
defined wavelengths selected on the Evaluation page are available.
Band Automatically calculated band maximum between the wavelengths x1 and x2

amplitude
Area Area under the band between the wavelengths 1 and 2
When enabling the Correction option, a baseline correction of the bands is
performed.
Corrected Baseline-corrected bands at a wavelength
band Corrected band x Enter the wavelength of the bands.
Range begin x1 and Range end x2 Enter the wave lengths between which
the baseline correction takes place.
Derivative 1st to 4th derivative of the spectrum between the wavelengths x1 and x2
Wavelength 1 and 2 Enter the wavelength range for the analysis of the
derivative
Order Order of the derivative
Supporting points Select the number of support points for the derivative
with the integrated smoothing filter
Range Select a value for the calibration (from 0 to
maximum, from 0 to minimum or from min. to max.)
7.1.5 Spectrum – Settings / Sample sequence
sequence
the accessories.
7.1.6 Spectrum – Settings / Archive automatically
7.1.7 Spectrum – Settings / Note
A note on the method may be optionally entered on the Note page.
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Spectrum module
7.2 Performing measurements in the Spectrum module
Button Description

sample]
one sample defined.
Measure a reference

measurement.
Execution
with samples.

displayed in the document window. During the measurement, the remaining
measurement time is displayed.
77
Spectrum module
Button Description
Stop measurement
processed further.
[Stop The current measurement step is stopped and then the processing

The Sequenz editieren window appears with the sequence stored in the method.
 Confirm the sequence with [OK].
Append individual Regardless of the sequence configured, single samples of sample type Probe may be
[Measure sequence].
switch to the Measure worksheet.
78
Spectrum module

Attention If additional worksheets besides the worksheet Measurement are already available in
the document, these are not updated during a re-measurement. Empty values may not
be re-measured.
When using cell changers or APG the sample to be re-measured must be placed in the
same position in the accessories. The position in the accessories can be displayed in the
sample table on the worksheet.
7.3 Displaying, analyzing and editing results in the Spectrum module
This section explains the particularities of the results display and analysis in the
Spectrum module. Please read up on the general structure of the document windows in
section "The document window" p. 15.
Document window in the Spectrum module with several spectra

1 Measurement worksheet 3 Measured values spreadsheet
2 New worksheet worksheet 4 Spreadsheet with data analysis
Measurement worksheet The Measurement worksheet includes the original data. The spectra are displayed in
the upper half of the worksheet.
The Measured values spreadsheet in the lower half of the worksheet contains the
sample table in the order of the measurements and the table with the digital measured
values. The additional spreadsheets include the data analyses already defined in the
method. These analyses can be amended and edited without changing the original
measured values.
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Spectrum module
New worksheet worksheet The New worksheet worksheet is created after clicking on the menu item with the same
name in the Measurements worksheet. The original data is first copied to the new
worksheet and can then be edited, e.g. by calculating a new baseline, a normalization or
a derivative (→ "Editing spectral data" p. 82).
Sample table The Measured values sample table in the lower part of the worksheet can be configured
freely. After right clicking on the table header the following parameters can be selected
in the context menu:
 Color
 Sample name
 Sample type
 Date and time of the measurement
 Temperature for measurements with Peltier-tempered accessories
 Notes
The individual measurements for each sample are listed adjacent in a separate table.
7.3.1 Display options for the results in the Spectrum module
The following functions are available for display in the Spectrum module:
 Scaling curves
These functions are described in the section "Show spectra and measurement curves on
the worksheet" p. 19.
7.3.2 Analyzing spectral data
analyses always relate to the measured values of the respective worksheet. If the
measured values are edited, the analysis on this worksheet is adjusted accordingly.
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Spectrum module
Display of the analysis The analysis are output in their own spreadsheet in the bottom part of the worksheet.
Worksheet with peak list
Edit analysis Each analysis function can only be used once in a worksheet. Thus a worksheet cannot,
for example, contain several peak lists. However, it is possible to edit the analysis
parameters and thus adjust the analysis in a spreadsheet.
Execution
 To edit an analysis, click on Settings in the spreadsheet of the analysis and change
the input parameters in the dialog window.
Delete analysis Analyses that are not required may be deleted.

Execution
Peak list An automatic peak search may be started with the Peaks function.
Execution
 Select Evaluation  Peak list on the worksheet.
 Configure the parameters in accordance with section "Spectrum – Settings /
Evaluation" p. 73.
 The peak list is displayed.
Values at defined The Values of def. wavelengths function compiles measured values of selected
wavelengths wavelengths in a table. The measured values can then be used further in a formula.
Execution
 Select Evaluation  Values of def. wavelengths on the worksheet.
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Spectrum module
 Select the wavelengths in accordance with section "Spectrum – Settings /

Evaluation" p. 73.
 The value table is displayed.
Formula With the function Formula, selected measured values of a spectrum results of the peak
search are linked in a formula. The values must already have been generated with the
analysis functions Peaks or Values of def. wavelengths.
Execution
38).
Tip
This function can only be applied to the measured values of the same spectrum. The
Formula function in the data analysis permits the linking of measured values from
different spectra (→ Chapter "Editing spectra data" / "Formula" p. 86).
calibration data to be loaded from other files and use them for a quantitative analysis.
Execution
 Select Evaluate  Calibration on the worksheet.
 Click on [Open] and select the file with the calibration curve.
 The Calibration spreadsheet with the current analysis is shown.
Attention When using the calibration from a different document only the calibration parameters
will be loaded. The calibration can therefore not be combined and re-calculated with
standards from the current document.
7.3.3 Editing spectral data
Run data editing The function Data handling allows for key operations to be performed during the
spectral analysis. The processing is logged on the audit trail of the document window. If
several spectra are overlayed in a window, the operations are applied to all spectra.
Data is only ever edited in the copy of the measured values in the New worksheet. The
data are saved.
Execution
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Spectrum module
this tab.
Execution
Trim When trimming a spectrum, select a spectral section. The values outside the selected
area are deleted.
Execution
 Select Data handling  Cut out on the worksheet.
The cursor changes to a vertical line with the label "Range".
 Click on the left or right limit of the x-coordinate range to be cut out and with the
mouse button held down move the cursor to the opposite limit.
Alternatively, enter the range limits in the window Cut out spectra in the fields
Begin and End.
 The spectral representation is updated. Now only the selected area of the
measurement graph is visible.
Time window When time-controlled multiple measurements (interval measurements, → "Spectrum -

Settings / General" p. 72) are taken, a three-dimensional spectral group with the
coordinates wavelength, absorbance and/or transmission and time will result. With a
time window a cut is placed through a group of graphs at a defined wavelength. The
result is a curve with the time-related change of the measured value at the selected
wavelength. This kinetic curve is displayed in a new document window of the Kinetics
module and can be analyzed and further edited there.
Execution
 Select Data handling  Time cut on the worksheet.
 In the input field enter the point on the x-axis (here wavelength) and click on the
corresponding wavelength in the spectrum.
 The resulting kinetic curve is displayed in a new document of the Kinetics
module.
Temperature cut When a spectra measurement with Peltier-tempered accessories is carried out in
Variable temperature mode, a three-dimensional spectral group with the coordinates
wavelength, and temperature will result. When a temperature cut is performed, a
section is applied to a spectral group at the selected wavelength. The result is a
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Spectrum module
absorbance/transmission graph in which absorbance or temperature is plotted against

the temperature. This graph is displayed in a new document window of the
Thermometry module and can be analyzed and further edited there.
Execution
 Select Data handling  Time cut on the worksheet.
 In the input field enter the point on the x-axis (here wavelength) and click on the
corresponding wavelength in the spectrum.
 The temperature graph is displayed in a new document of the Thermometry
module.
Offset An offset moves the entire spectrum evenly along the y-axis by adding a constant to the
measured values. By entering positive or negative values a movement is possible in both
directions. This allows for interferences to be simulated or compensated.
Execution
 Select Data handling  Offset on the worksheet.
 The spectrum is corrected with the offset value and the display updated.
Factor With the function Factor the measured values of the spectrum are multiplied by a
constant. This expands or compresses the spectrum. The multiplication of an
absorbance spectrum with a constant theoretically corresponds to a change of the layer
thickness of the sample and/or the concentration.
Execution
 Select Data handling  Factor on the worksheet.
 The spectrum is multiplied with the factor and the display updated.
Smoothing The spectrum is smoothed using the Savitzky-Golay method.

Execution
 Select Data handling  Smooth on the worksheet.
 In the dropdown list Base points, select the number of points to be considered for
smoothing in accordance with Savitzky-Golay.
 The spectrum is re-calculated and the display updated.
Derivative For the spectrum, the derivation of the 1st to 4th order with integrated constant
smoothing over 5 points and variable number of points for the derivation filter (n
points) according to Savitzky-Golay is calculated. The derived spectrum can suppress
background signals overlayed on the measurement spectrum and more significantly
highlight the specific absorbances.
A spectrum can be derived several times. However, the algorithm is specifically adapted
to the derivative type; therefore the fourth derivative does not result in the same y-
coordinate values as four times the first derivative. Therefore, the same method should
always be used for the quantitative analysis.
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Spectrum module
Especially the derivative of a higher order always results in the "coarsening" of the
measurement graph. Therefore, dependent on the width of the measurement graph
structure, always select the highest possible number of support points. Smoothing
beforehand to suppress the statistical noise is sometimes also recommended. Note that
the value range will be reduced in accordance with the selected number of base points
(n) by the number of values (n-1)/2+2 on both ends of the spectra. To reduce this effect
a prior interpolation is recommended.
Execution
 Select Data handling  Derivative on the worksheet.
 In the Derivative list, select the order of the derivative.
 In the Base points list, select the number of support points to be used for the
derivative with integrated smoothing filter.
 The derivative is calculated and the display updated. In the audit trail the order
of the derivative and the number of support points are documented.
Normalize In normalization, the spectrum is multiplied with a factor k to achieve a specified y-

coordinate value at a wavelength or peak.
Execution - Normalization at a wavelength
 Select Data handling  Normalization  One point on the worksheet.
The cursor changes to a continuous vertical line with the label "W".
 In the graphic, click on the required wavelength.
 Edit the wavelength in the input window, if necessary, and enter the Normalization
value.
Execution - Normalization at a peak
 Select Data handling  Normalization  Maximum in band on the worksheet.
 Click on the left or right limit of the peak and with the mouse button held down
move the cursor to the opposite limit.
 Edit the wavelengths of the peak limits in the input window, if necessary, and enter
the Normalization value.
Interpolate The function Interpolation permits the interpolation of a spectrum by changing the
measuring point distance. The function can be used to gain intermediate values for a
more precise peak determination. The interpolation is optionally linear or with a cubic
spline.
Execution
 Select Data handling  Interpolation on the worksheet.
 In the dropdown list Calculation, select the calculation type.
 In the Step width field, enter the new measuring point distance. The new
measuring point distance must be smaller than the original one.
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Spectrum module
Mean value spectrum The function Average spectrum calculates the mean value spectrum from a group of
graphs.
Execution
 Select Data handling  Average spectrum on the worksheet.
 The mean value spectrum is calculated and the display updated.
Baseline correction With the Baseline correction, a rising or falling baseline of a band can be corrected.
The linear function of the baseline is determined by two points in the graph.
Execution
 Select Data handling  Baseline correction on the worksheet.
 Click with the mouse on the first point in the spectrum, drag the cursor with the
mouse button held down to the second point and release.
 In the input window edit the two points (start point and end point) for the linear
function, if necessary, and confirm with [OK].
Auto baseline correction With the auto baseline correction a rising or falling baseline of a band is automatically
corrected. The linear function of the baseline is placed through the minima on both
sides of the peak.
Execution
 Select Data handling  Auto baseline correction on the worksheet.
Conversion to This function permits the conversion of a spectrum from absorbance to transmission
transmission [%T] / [%T] and vice versa.
absorbance
Execution
 Select Data handling  Convert to transmittance [%T] (Convert to absorbance) on
a new worksheet.
 The spectrum is converted and the display updated.
Formula The Formula function permits

 The addition/subtraction and multiplication/division of all measured values of a
spectrum with a constant
An absorbance spectrum can e.g. be multiplied with a constant. This corresponds
theoretically to a change of the layer thickness or concentration change of the
sample.
 Linking spectra arithmetically to each other
e.g. is the addition of the spectra of two (or several) known substances is useful, if
the total spectrum of these substances is, for example, to be created for comparison
purposes.
86
Spectrum module
A condition for linking the spectra to each other is that the spectra are overlayed
together in a window. Spectra from different files can be combined with Copy/Paste in
a new window (→ "Combining measuring results from different documents" p. 87).
Execution
 Select Data  Handling on the worksheet.
 Enter the function in the formula editor (→ "Using formulas" p. 38).
The spectra can be selected as arguments in the list Choose spectra and inserted
into the formula by clicking on .

 The result is calculated and the display updated.
7.3.4 Combining measuring results from different documents
Copy and insert data This function permits the overlaying and joint editing of spectra from different
documents. To this end the spectra are copied from their original file to a new
document. The spectra may have different wavelength ranges and measuring point
distances.
Execution
 In the Spectrum module, open all files where the required spectra are located (e.g.
Spektrum1.auv and Spektrum2.auv).
 Open a new document by clicking on [Spectrum].

 In the first document (Spektrum1.auv), activate all spectra that need to be copied
(→ "Hide curve / (De)activate samples" S. 22).
 In the first document (Spectrum1.auv), select File  Copy.
 Change to the new document and select there File  Paste.
 From the second document copy the required spectra in the same manner to the
new document.
 The spectra in the new document can now be edited.
7.4 Example measurement for the Spectrum module
As an example a spectral measurement in the range from 250 to 700 mm is used. The
Holmium standard from the standard filter set of Hellma can be used as sample if
available. The measurement is performed with the standard cell holder:
 Create document in module.

Click on [Spectrum] in the launch bar or select Modules Spectrum of the main
window.
87
Spectrum module
Define method  Click on [Method settings].

parameters
 Enter the parameters on the pages of the Method window.
 Exit the parameter input by clicking on [OK] in the Spectrum - Settings window
Parameter entries Enter the parameters as shown in the screenshot.

General page:

Evaluation page: Make no entries.
Sample sequencepage:
88
Spectrum module
 Enter "Reference" in the Name field of the reference line and confirm with ENTER.
 Click on [Add samples] once again and add one sample to the end of the sample
table:
 In the second line of the sample table, enter "Holmium" as the name and confirm
with ENTER.

Note page: Enter "Example for spectra scan".

measurement
 Insert the reference sample (filter F0) and click on [Yes] to start the measurement.
displayed.
 Insert sample1 (filter F1) and click on [Yes] to start the measurement.
 The Holmium spectrum appears in the document window.
89
Spectrum module
Example of a spectral scan
90
Kinetics module
8 Kinetics module
In the Kinetics module, time-related changes of the measured values at a wavelength
are recorded and analyzed.
Open the module Kinetics Execution

 Open the Kinetics module by clicking on the button in the launch bar of the main
window:
Alternatively, select the item Module  Kinetics in the menu.

assigned to the Kinetics module.
8.1 Method configuration in the Kinetics module
This chapter includes all configurations that can be made for a method in the module
Kinetics. Please also pay attention to the notes in chapter "What you should know about
ASpect in advance" p. 11.

 Configure the method on the pages of the window Kinetics - Settings.
 The method parameters are output on the left side of the document window on
the Method tab. The buttons for starting the measurement appear.
8.1.1 Kinetics - Settings / General
On the General page, the basic measurement configurations are made.
91
Kinetics module
Kinetics – Settings / General window
Option Description
Measurement Select transmission or absorbance as y-axis display

mode
Wavelength Enter the wavelength at which the kinetics measurement is performed
Integration Time for recording a measuring point

time
0.2; 0.5; 1; 2; 4 nm
been installed.

The preset lamp change at 320 nm guarantees an optimal distribution of
92
Kinetics module
Time cycle Enter times for the kinetics measurement.
Option Description
Measuring Enter total measuring time

time
Interval Enter the time between the recording of two consecutive measured values
time
Delay time Enter the delay time between the measuring start and actual start of the first
measurement
Display Select the time unit for the x-axis of the graphical representation
time in
chart
Cyclic for For cell changer or APG
Sample The entire reaction-kinetic measurement is performed
in one sample and then the measurement started in
the next sample. All samples in the sequence are
processed consecutively in this manner.
This measurement is suited for quick reaction kinetics
if the reaction is started just before the measurement
in each case. On the Accessory page, the Manual start
option must be enabled.
Batch In this mode, the sample measurements are staggered
in time. Within an interval, all samples of the sequence
are measured, starting with the 1st sample. The next
interval then re-starts with measuring the first sample.
In this manner the measurement is continued until the
last interval.
One batch of the cell changer/APG is measured at a time. Once all
the samples are replaced in the accessories and the next batch is measured.
Measurement If the Measurement break option is enabled, the measurement is interrupted once the
cancellation absorbance or transmission value entered has been reached (depending on ordinate
unit selected). It is possible to select in the list whether the value represents the upper or
lower limit.
8.1.2 Kinetics – Settings / Accessory
The parameters in the Accessory page depend on the installed accessories. The
configurations have been described in section "Using accessories in SPECORD PLUS"
p.40.
8.1.3 Kinetics – Settings / Evaluation
On the Evaluation page, values can be determined at defined points in time and linked
to a formula.
93
Kinetics module
Kinetics – Settings / Evaluation window
Select measured values at Measured values at selected points in time are displayed in the sample table and can be
defined points in time used for the analysis in a formula or in the calibration.
Execution
 Click on the Values of defined times option to select the measured values.
 Click on , enter the desired time in the input field and transfer it to the list with
.
Select points in time from the measuring graph
 To delete the value select it in the list and click on .
Formula If the Formula option is enabled, the measured values are linked in a formula (→ "Using
formulas" p. 38).
The following functional arguments can be used:
 Measured values at selected points in time (e.g. "t 02:00")
 Slope of kinetic curve
The slope of the kinetic curve is first calculated over the entire measuring duration. After
Tip
completion of the measurement, the points of time for the range to be analyzed can be
defined (→ "Range of the slope calculation" p. 101).
8.1.4 Kinetics – Settings / Calibration
On the Calibration page, you can define an analysis with calibration. The Calibration
checkbox must be enabled for this purpose.
94
Kinetics module
Kinetics – Settings / Calibration
Calibration configuration Option Description

series. To this end standard samples and their
concentrations must be recorded in the sample
sequence.
Load from file Load calibration parameters from an existing results
file.

y=B*x
y=A+B*x
y = A + B * x + C * x2
Pathlength Enter the pathlength of the cell.

This value is for information only and is saved with the method parameters.
above or below this limit value are identified in the results table.
Ordinate values The Kinetics module can use different values from the time-related measuring graph for
the calibration:
Ordinate value Explanation
Slope Slope of the entire kinetic curve from the start to the end time of the
measurement. The range may be limited after completion of the
measurement.
Maximum Maximum of kinetic chart between Start time and End time
between t1 and
t2
Formula Result of a formula calculation

After clicking on the formula editor appears (→"Using formulas" p.
38). As arguments for the formula, the time values that have been
selected on the Evaluation page and the slope of the kinetic curve are
available.
95
Kinetics module
Lineweaver Reciprocal reaction speed 1/v

Burk
8.1.5 Kinetics – Settings / Sample Sequence
sequence
the accessories.
8.1.6 Kinetics – Settings / Archive automatically
8.1.7 Kinetics - Setting / Note
A note on the method may be optionally entered on the Note page.
8.2 Performing measurements in the Kinetics module
Button Description

sample]
one sample defined.
96
Kinetics module
Measure a reference

SPECORD PLUS" p.40.

measurement.
 The reference value at the analysis wavelengths is taken.
All subsequent measurements are corrected with this reference value until the
measuring parameters are modified by a new method configuration or the next
reference measurement takes place.
Execution
with samples.

Button Description
Stop measurement
processed further.
[Pause] The measurement stops immediately without displaying the sample

sequence still pending and is continued by clicking on [Weiter].
The current measurement step is stopped and then the processing
[Suspend interrupted.
sequence] The current measurement step is stopped and then the processing
interrupted. The measurement pause may be used to view and edit the
97
Kinetics module

 Confirm the sequence with [OK] .
Append individual Regardless of the sequence configured, single samples of sample type Sample may be
[Measure sequence].
8.3 Displaying, analyzing and editing results in the Kinetics module
This section explains the particularities of the results display in the Kinetics module
window. The general structure of the document windows is detailed in section "The
document window" p. 15.
98
Kinetics module
Document window in Kinetics module with several kinetic curves

1 Measurement worksheet 4 Calibration spreadsheet (analysis)
2 New Worksheet worksheet 5 Kinetic curves (ordinates and time of
3 Measured values spreadsheet measurement)
The spreadsheet Measured values located in the lower half contains the worksheet in
the order of the measurements and the table with the digital measured values. The
additional spreadsheets include the data analyses already defined in the method. These
analyses can be amended and edited without changing the original measured values.
New worksheet The New worksheet worksheet is created after clicking on the menu item with the same
name in the Measurements worksheet. The original data are first copied to the new
worksheet and can then be edited.
 Graph color
 Sample name
 Sample type
 Slope
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Kinetics module
 Slope from / slope to (range limits of the slope)

 Temperature for measurements with Peltier-tempered accessory
 Notes
8.3.1 Display options for the results in the Kinetics module
The following functions are available for displaying the kinetic curve:
 Scaling curves
Time display for kinetic The display of the x-coordinate in the kinetic curve can be changed between hours,
curve minutes and seconds.
Execution
 Select Options Show time in the chart as and enable the unit in the submenu.
8.3.2 Evaluate kinetic curves
All data analyses available in the method can also be subsequently applied to the
measured values. The functions have been compiled in the menu Evaluation that is
available on every worksheet. The analyses always relate to the measured values of the
respective worksheet. If the data is edited, the analysis on this worksheet is adjusted
accordingly.
Display of the analysis The analysis are output in their own spreadsheet in the bottom part of the worksheet.
100
Kinetics module
Kinetics chart with display of the values at defined points in time
Edit analysis Each analysis function can only be used once in a worksheet. However, it is possible to
edit the analysis parameters and thus adjust the analysis in a spreadsheet.
Execution
 To edit an analysis, click on Settings in the spreadsheet of the analysis and
change the input parameters in the dialog window.
Delete analysis Analyses that are not required can be deleted.

Execution
Range of the slope The slope is initially calculated over the entire kinetic curve. After completion of the
calculation measurement, this range can be limited and only a section of the reaction kinetics,
ideally the linear part, is used for the calculation.
Execution
 If the slope is only calculated for individual samples, highlight these samples on the
worksheet (→ "Display options for the results in the Kinetics module" S. 100).
 Select Evaluation Range for slope calculation on the worksheet.
 Enter times for the start and end of the linear range.
 In the list select the samples for which the slope will be calculated.
Option Description
For all measurements The calculation is performed for all samples
Only for highlighted The calculation is only performed for the previously highlighted
measurements samples.
101
Kinetics module
 The value of the slope in the measured values table is updated. Any analyses
are also re-calculated.
Determine range for slope calculation under Analysis
The slope calculation range may also be edited in the sample table on the Measured
values spreadsheet:
 Click on the Slope from and Slope to table cells to edit the range limits.
Edit the slope calculation range in the sample table
Values of def. times The Values of defined times function compiles the measured values of selected times in
a table. The measured values can then be used further in the formula analysis.
Execution
 Select Evaluation  Values of defined times on the worksheet.
 Select the wavelengths in accordance with section "Kinetics – Settings / Evaluation"
p. 93.
Formula With the Formula function, measured values at defined times or the slope can be linked
in a formula. This is possible because these values have already been created with the
Values of def. times analyses.
Execution
38).
calibration data to be loaded from other files and used for a quantitative analysis.
102
Kinetics module
Execution
 Select Evaluate  Calibration on the worksheet.
 Click on [Open] and select the document with the calibration graph.
 The Calibration spreadsheet with the current calibration graph and the
calculated calibration parameters is displayed.
Attention When using the calibration from a different document only the calibration parameters
will be loaded. The calibration can therefore not be combined and re-calculated with
standards from the current document.
Calibration graph display
The following is shown on the Calibration spreadsheet:
 Source of the calibration graph (measured in this document or loaded from a file)
 Regression model
 Regression coefficients A, B and C if calculated in the regression model
 Pathlength of the cell (for information only, not used in the calculation)
 Date/time of the measurement
Note:
Measured results outside of the calibrated range are placed in brackets in the sample
table, e.g. "(45.3)".
8.3.3 Editing kinetics data
Run data editing In the Kinetic module, the following functions may be executed in the Data handling
menu:
 Trim kinetic curve
 Smooth kinetic curve
The processing is logged on the audit trail of the document window. If several kinetic
curves are overlayed in a window, the operations are applied to all curves.
Data is only ever edited in the copy of the measured values in the New worksheet. The
data are saved.
Execution
 In the measured values worksheet, click on [+New worksheet].
 In the Data handling menu, select the desired function.
this tab.
103
Kinetics module
Execution
Trim The Cut out spectra function allows for the selection of a kinetic curve and deletes the
values outside the selected range.
Execution
 Select Data handling  Cut out spectra on the worksheet.
Alternatively enter the range limits in the window Trim in the fields Start and End.
 After clicking on [OK], the display is updated. Now only the selected area of the
kinetic curve is visible.
Smoothing The kinetic curve is smoothed using the Savitzky-Golay method.

Execution
 Select Data handling  Smooth on a new worksheet.
 In the dropdown list Base points select the number of points to be considered for
 The kinetic curve is re-calculated and the display updated.
8.4 Example measurement for the Kinetics module
As an example a kinetics measurement is demonstrated at 500 nm over a time period of

30 s. The demonstration measurement can be performed with an empty sample space.
 Create document in module.

Click on [Kinetics] in the launch bar of the main window or select Modules 
Kinetics.
104
Kinetics module

parameters
 Enter the parameters in the method window.
 Exit the parameter input by clicking on [OK] in the Kinetics - Settings window and
return to the document window.
Parameter entries General page:

Evaluation page: Make no entries.
105
Kinetics module
 Click on [Add samples] once again and add a sample to the end of the sample
table:
Automatic save page: Make no entries.

Note tab: Enter the note "Example for kinetics".

measurement
 Insert the reference sample and click on [Yes] to start the measurement.
The reference measurement is performed.
 The prompt to measure sample 1 is displayed. Click on [Yes] to start the
measurement.
 The kinetic curve is shown in the document window.
106
Kinetics module
Example of kinetic measurement recording
107
Thermometry module
9 Thermometry module
In the Thermometry module, temperature-resolved measured curves can be recorded
and analyzed with a Peltier-tempered accessory. The following analyses are available:
 DNA melting point determination
 Output of measured values at selected temperatures
 Linking of the measured values in a formula
Opening the Execution

Thermometry module
 Open the Thermometry module by clicking on the button in the launch bar of the
main window.
Alternatively, select the item Module  Thermometry in the menu.
assigned to the Thermometry module.
9.1 Method configuration in the Thermometry module
This section includes all configurations that can be made for a method in the
Thermometry module. Please also pay attention to the notes in chapter "What you
should know about ASpect in advance" p. 11.

 Configure the method on the pages of the Thermometry - Settings window.
9.1.1 Thermometry – Settings / General
In the Thermometry - Settings / General window, the basic measurement

configurations are made.
108
Thermometry module
Thermometry - Settings / General window with configuration of the measurement parameters
Option Description
Measurement Select transmittance or absorbance as ordinate display

mode
Wavelength Select wavelengths for the analysis
Integration Select the time for recording a measuring point.

time

0.2; 0.5; 1; 2; 4 nm
been installed.

Measurement The temperature-dependent measurement can be ended when a defined measurement

cancellation value (absorbance or transmission) is reached.
Option Description
Measurement Enable the checkbox when the measurement is to be canceled at a

break predefined limit value.
Value Select the limit value for the cancellation.
List Select approximation to limit value: Upper limit or Lower limit.
109
Thermometry module
9.1.2 Thermometry – Settings / Accessory
In the Thermometry – Settings / Accessory window, use the settings for the
temperature curve parameters.
Thermometry – Settings / Accessory window with the temperature curve settings
Parameter settings Option Description
Synchronous Only available when 2 cell changers are used:

If enabled the two cell changers move synchronously. A separate
reference can be used for each sample.
If disabled, the first samples in the reference channel are
measured followed by the samples in the measuring channel
(offset mode).
See also section "Cell changer" p. 42
Manual start Only available when the cell changer is used:
Every sample measurement must be started by pressing a button.
Precision of Select the accuracy of the temperature control in the accessories.
temperature
Cell sensor active During the sample measurement, the actual temperature is
measured in the cell and assigned to the measured value. To do
so, the temperature sensor must be installed in the cell.
Temp. after When this option is active the entered temperature is approached
measurement and held after the last optical measurement; otherwise the last
temperature in the temperature table is held.
Temperature values Use [Setup] to open the window for entering the temperature
values.
Entering the temperature After clicking on [Setup], the window Variable temperature mode appears. The
progression parameters of the temperature stages can be edited individually in the table rows or
defined automatically using the checkboxes and input fields.
Option Description
[Insert] Enter a new temperature step after the highlighted table row
[Delete] Delete the highlighted table row
110
Thermometry module
Gradient from Automatically generate a temperature gradient from a highlighted table

marked row. Starting with the start temperature of the highlighted table row
position further temperature stages are inserted into the table in steps until the
final temperature has been reached.
Diff. [°C] Temperature difference of a temperature stage
to °C Final temperature of the gradient
[Add] Add temperature gradient at the highlighted
position
Waiting time Waiting time from reaching the target temperature to starting the optical
for all measurement
With [Set], the temperature entered is transferred to all temperature
stages of the table.
Note: Regardless of the set waiting time, the optical measurement is
carried out no less than 5 s after reaching the temperature within the
agreed temperature accuracy range in the block. The waiting time is added
to this 5 s period.
[Undo] Reverse the last action
Optical Yes Perform an optical measurement at temperature

measurement step
No No optical measurement takes place at the
temperature step.
Variable temperature mode window for setting the temperature curves
111
Thermometry module
9.1.3 Thermometry – Settings / Evaluation
In the Thermometry – Settings / Evaluation window, the following analyses may be

selected:
 Determination of DNA melting point
 Output of measured values at selected temperatures
Thermometry – Settings / Evaluation window
Determining the melting When the Determination of DNA melting point option is activated, the melting point of
point the DNA melting curves is determined.
Determine peak By enabling the Determine peaks checkbox, extreme values are automatically searched
for based on specified criteria. The peaks are highlighted in the temperature curve after
the measurement and output to a separate analysis table. Peaks can be used in a
formula.
Option Description
List of the complete spectrum Search for all peaks in the measuring range
that meet the criteria below.
only in defined ranges of spectrum Enter different spectral ranges in which
peaks are searched for.
Type of peak Select the type of peak: Minima, Maxima or Minima and Maxima.
112
Thermometry module
Show peaks Absolute limit for displaying the peaks

only Peaks are only displayed if they are above or below the ordinate value
entered.
Threshold Height of peak taking baseline into consideration

Difference of peak height between local minimum/maximum and the
subsequent neighboring local maximum/minimum.
Selecting measured values Measured values at selected temperatures are displayed in a table and can be used with
at defined temperatures a formula for the analysis.
Click on the Values of defined temperatures option to select the measured values. Then
open an input field with and consecutively enter the desired temperatures.
Using formulas If enabled, the measured values are linked in a formula (→ "Using formulas" p. 38).
The values entered above can be used as functional arguments:
 Absorbance/transmission of peaks in selected temperature ranges
 Measured values at defined temperatures
9.1.4 Thermometry – Settings / Sample Sequence
Requirements for sample  When using cell changers the samples are assigned to the sample spaces in the
sequence accessories.
9.1.5 Thermometry – Settings / Archive automatically
9.1.6 Thermometry – Settings / Note
A note on the measurement may be optionally entered on this page.
113
Thermometry module
9.2 Performing measurements in the Thermometry module
Button Description

sample]
one sample defined.
Measure a reference
[Reference] A separate reference measurement is required if the sample sequence
does not start with a reference measurement or no reference is available
that matches the current method parameters.
[Set Approach the start temperature for the measurement on the cell block.
Temperature]

SPECORD PLUS" p.40.

measurement.
 The reference measurement is performed at the start temperature.
Execution
 Place the first sample of the sequence in the cell holder or fill cell changer with
samples.
114
Thermometry module
Button Description
Stop measurement
processed further.
[Suspend The current measurement step is stopped and then the processing

[Measure sequence].
115
Thermometry module
9.3 Displaying, analyzing and editing results in the Thermometry module
This section explains only the particularities of the results display in the Thermometry
window. Please read up on the general structure of the document windows in section
"The document window" p. 15.
Document window in the Thermometry module

1 Measurement worksheet 3 Measurement values spreadsheet
2 New worksheet worksheet 4 DNA melting point spreadsheet (analysis)
The Measurement spreadsheet in the lower half of the worksheet contains the sample
table in the order of the measurements and the table with the digital measured values.
The additional spreadsheets include the data analyses already defined in the method.
These analyses can be amended and edited without changing the original measured
values.
New worksheet worksheet The New worksheet worksheet is created after clicking on the menu item with the same
name in the Measurements worksheet. The original data is first copied to the new
worksheet and can then be edited, e.g. by calculating a derivative (→ "Editing
temperature curves" p. 119).
 Position in the accessories when using a cell changer
 Color (graph color)
116
Thermometry module
 Batch when using cell changers

 Sample name
 Sample type
 Slope and range limits for the calculation
 Notes
9.3.1 Display options for the results in the Thermometry module
The following functions are available for displaying the temperature curve:
 Scaling curves
9.3.2 Evaluating the temperature curves
analyses always relate to the curves of the respective worksheet. If the data is edited, the
analysis on this worksheet is adjusted accordingly.
Display of the analysis The analyses are output in their own spreadsheet in the bottom part of the worksheet.
117
Thermometry module
Worksheet with DNA melting point
Edit analysis Each analysis function can only be used once in a worksheet. Thus a worksheet cannot,
for example, contain several peak lists. However, it is possible to edit the analysis
parameters and thus adjust the analysis in a spreadsheet.
Execution
 To edit an analysis, click on Settings in the spreadsheet of the analysis and change
the input parameters in the dialog window.
Delete analysis Analyses that are not required may be deleted:

Execution
Calculating the melting The melting point can be determined in a DNA melting curve.
point
Execution
 Select Evaluation  DNA melting point on the worksheet.
 An overview of the determined melting points is displayed.
Peak list function The Peaks function automatically determines and displays peaks in the temperature
curve.
Execution
 Select Analysis  Peak list on the worksheet.
 Configure the parameters in accordance with section.
 The peak list is displayed.
118
Thermometry module
Values of def. The Values of def. temperatures function compiles measured values of selected
temperatures temperatures in a table. The measured values can then be used further in a formula.
Execution
 Select Evaluation  Values of def. temperature on the worksheet.
 Select the wavelengths in accordance with section "Thermometry – Settings /
Evaluation" p. 112.
Create formula With the Formula function, selected measured values or results of the peak search of a
temperature curve are integrated in a formula. The values must already have been
generated with the Peak list or Values of def. temperatures analysis function.
Execution
38).
Selecting the range of the The slope is initially calculated over the entire temperature curve. After completion of
slope calculation the measurement, this range can be limited and only a section of the temperature curve,
ideally the linear part, is used for the calculation.
Execution
 If the slope is only calculated for individual samples, highlight these samples on the
worksheet (→ "Display options for the results in the Thermometry module" S. 117).
 Select Evaluation  Slope range on the worksheet.
 Enter times for the start and end of the linear range.
 In the list select the samples for which the slope will be calculated.
Option Description
Set to all measurements The calculation is performed for all samples
Set only to marked The calculation is only performed for the previously
measurements highlighted samples.
 The value of the Slope in the measured values table is updated. Any analyses
are also re-calculated.
9.3.3 Editing temperature curves
The Data handling function includes mathematical functions for editing the
temperature curves. The processing is logged on the audit trail of the document.
If several temperature curves are overlayed in a document, the operations are applied to
all charts. The data editing also affects existing analyses. The analyzed data are updated
when you open the corresponding spreadsheet.
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Thermometry module
Temperature curves are always edited only in the copy of the original data. The original
measured values are not modified by these operations but remain intact. When saving
the documents the edited and the original data are saved.
Execution
this tab.
Execution
Trim When trimming a temperature curve, a graph section is selected and the values outside
of the selected range are deleted.
Execution
 Select Data handling  Trim on the worksheet.
Optionally, enter the range limits in the Trim spectra window in the Start
temperature and End temperature fields.
 After clicking on [OK], the graphic is updated. Now only the selected area of the
measurement graph is visible.
Offset An offset moves the entire temperature curve evenly along the ordinate by adding a
constant to the measured values. By entering positive or negative values a movement is
possible in both directions.
Execution
 Select Data handling  Offset on the worksheet.
 The temperature curve is corrected with the offset value and the display is
updated.
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Thermometry module
Factor With the Factor function, all measured values of the temperature curve are multiplied by
a constant. This expands or compresses the curve along the y-axis.
Execution
 Select Data handling  Factor on the worksheet.
 The temperature curve is corrected with the offset value and the display
updated.
Smoothing The temperature curve is smoothed using the Savitzky-Golay method.

Execution
 Select Data handling  Smooth on the worksheet.
 In the Support points dropdown list select the number of points to be considered for
 The temperature curve is re-calculated and the display updated.
Derive For the temperature curve the derivation of the 1st to 4th order with integrated
constant smoothing over 5 points and variable number of points for the derivation filter
(n points) according to Savitzky-Golay is calculated.The derived curve can suppress
background signals overlayed on the measurement spectrum and more significantly
highlight the specific absorbances.
A curve can be derived several times. However, the algorithm is specifically adapted to
the derivative type, therefore the fourth derivative does not result in the same y-
coordinate values as four times the first derivative. Therefore, the same method should
always be used for the comparative analysis.
Especially the derivative of a higher order always results in the "coarsening" of the
measurement graph. Therefore, dependent on the width of the measurement graph
structure, always select the highest possible number of support points. Smoothing
beforehand to suppress the statistical noise is sometimes also recommended. Note that
the value range will be reduced in accordance with the selected number of base points
(n) by the number of values (n-1)/2+2.
Execution
 Select Data handling  Derivative on the worksheet.
 In the Derivative list, select the order of the derivative
 In the Base points list, select the number of support points to be used for the
derivative with integrated smoothing filter.
 The derivative is calculated and the display updated. In the audit trail the order
of the derivative and the number of support points are documented.
9.4 Example measurement for the Thermometry module
In the example, the melting point of a DNA sample is determined. For this purpose, a
temperature curve in a range between 25 – 90 °C is run. The measurement requires
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Thermometry module
Peltier-tempered accessories with a cell sensor. A suitable DNA sample or water can be
used as sample.

 Click on [Thermometry] in the launch bar or select Modules  Thermometry of
the main window.

parameters
 Enter the parameters on the tab of the Method window.
 Exit the parameter input by clicking on [OK] in the Thermometry - Settings
window and return to the document window.
Entry of parameters General page:
Accessory page:
Setting the temperature curve parameters:

 Open the Variable temperature mode window by clicking on [Setup].
 Click on the Temperature column and enter the value "25".
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Thermometry module
 Enter the value "1" in the Diff. [°C] field and "30" in the to °C field. Click on [Add].
 Repeat these entries for the value pairs "0.5 °C"/"80 °C" and "1°C"/"°C90".
 Confirm entries with [OK].
Evaluation page:
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Thermometry module
table:

Note page: Enter the note "Example of melting point determination".
Running the  Insert the reference sample and click on .

measurement
 When the message for starting the reference measurement appears, lick [Yes].
displayed.
 Insert sample 1 and click on [Yes] to start the measurement.
 The temperature curve appears in the document window.
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Thermometry module
Example of melting curve with melting point determined
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Colorimetry module
10 Colorimetry module
The Colorimetry module performs the colorimetric analysis of transmission spectra of
transparent solid or liquid samples and reflection spectra of non-transparent solid or
powdery samples.
The following parameters can be determined in the Colorimetry module:
Color spaces/Tristimulus  Tristimulus values X, Y, Z (as per EN ISO 11664)

values
 Tristimulus values x, y (as per EN ISO 11664)
 CIE lab, CIE luv (as per EN ISO 11664)
 Hunter lab
The following parameters can be selected to calculate the color coordinates:
 Viewing angle 2° and 10°
 CIE illuminant A and D65 according to EN ISO 11664
 CIE illuminant C
Color numbers  Degree of whiteness/yellowness index

 Platinum Cobalt acc. to ASTM D 5386-05 with linear regression over yellow index
acc. to ASTM E313-15
 Iodine with calibration function via CIE lab values
 Gardner according to DIN ISO 4630
 Saybolt calculated from CIE lab values acc. to ASTM D156, D6045
Open the Colorimetry Execution

module
 Open the Colorimetry module by clicking on the button in the launch bar of the main
window:
Alternatively, select Module  Colorimetry in the menu.
assigned to the Colorimetry module.
10.1 Method configuration in the Colorimetry module
This chapter includes all configurations that can be made for a method in the
Colorimetry module. Please also pay attention to the notes in chapter "What you should
know about ASpect in advance" p. 11
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Colorimetry module
 Configure the method on the pages of the Colorimetry - Settings window.

10.1.1 Colorimetry - Settings / General
In the Colorimetry - Settings / General page, the basic measurement configurations are
made.
Colorimetry - Settings / General window with configuration of the measurement parameters
Option Description
Parameter Click on [Set] to set the measurement parameters for the spectral
according to EN range, the measuring point distance and the spectral width required by
ISO 11664 EN ISO 11664.
Measurement Select Transmittance or Reflectance as ordinate display.

mode
Range Enter the wavelength range for the measurement in the input fields.
Adjustment range: For the colorimetric analysis, a spectral range of at
least 380 – 780 nm must be captured.
Measuring points Select distance between measuring points.
Speed Select measuring speed.
Integration time Time for recording a measuring point

This value is calculated automatically from the measuring point
distance and measuring speed.
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Colorimetry module

0.2; 0.5; 1; 2; 4 nm
* The 0.2 nm slit is only displayed in the SPECORD 210/250 PLUS where it has
actually been installed.
Lamp change at Select wavelength of change from UV lamp to Vis lamp

The preset lamp change at 320 nm guarantees an optimal distribution
of energy across the entire wavelength range of the spectrometer. If
you are only working in UV or Vis range, you may only use the
corresponding lamp with the options UV only or Vis only.
10.1.2 Colorimetry – Settings / Accessory
The parameters in the Accessory page depend on the installed accessories. The
configurations have been described in section "Using accessories in SPECORD PLUS"
p.40.
10.1.3 Colorimetry – Settings / Color values
On the Color values page, the color coordinates and color numbers to be calculated are
selected.
Colorimetry – Settings / Color values window
Selection of color The following color coordinates can be calculated in ASpect UV according to EN ISO
coordinates 1164:
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Colorimetry module
 Tristimulus values for X, Y, Z

 Proportion of tristimulus values x, y
 CIE lab
 CIE luv
 Hunter lab
Parameter The color coordinates can be determined with two viewing angles:
 2°
 10°
The following illuminants are available:
 Standard Illuminant A according to EN ISO 1164
 Standard Illuminant D65 according to EN ISO 1164
 Illuminant C
Multiple color coordinates and parameters can be selected for the analysis of a
measurement.
Color numbers The following color numbers can be calculated:

 Degree of whiteness, yellow index
 Platinum Cobalt acc. to ASTM 5386, with lin. regression via yellow index acc. to
ASTME 313
 Iodine with calibration function via CIE lab values
 Gardner according to DIN ISO 4630
 Saybolt calculated from CIE lab values acc. to AST D156, D6045
Pathlength In order to determine the color values, color coordinates and the color numbers derived
from these, a transmission or reflection value is required for the normalized pathlength
of 1 cm (10 mm).
 For transmission measurements, select the active pathlength in the direction of the
beam for the sample or cell used from the Pathlength list or enter the value directly
in the list field.
 For reflection measurements of impermeable surfaces, the value must be normalized
at 10 mm (1 cm).
Attention In the standards for determining the color numbers Platinum Cobalt (Hazen), Iodine and
Gardner, it is important to remember that an application may only take place if the
absorbance characteristics or the color value of the sample are similar to the
corresponding standard. The color value is defined by its spectral color. In order to
guarantee the validity of a transparent standard, e.g. water, the definition of an Epsilon
environment around the achromatic point is required. Both parameters are accordingly
predefined in an Ini-file for the specified color values, taking both metrological and
color-specific aspects into consideration. If these limits need to be changed, open the
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Colorimetry module
colcount.ini file in an editor at ProgramData\Analytik Jena\ASpect UV. Change the values

to meet your requirements and save the file again.
10.1.4 Colorimetry – Settings / Sample Sequence
On this page, a sample sequence can be prepared and saved with the method
parameters (→ "Sample sequences" p. 32).
Requirements for sample  When using cell changers and APG, the samples are assigned to the sample spaces in
sequence the accessories.
10.1.5 Colorimetry – Settings / Archive automatically
10.1.6 Colorimetry – Settings / Note
A note on the measurement may be optionally entered on the Note page.
10.2 Performing measurements in the Colorimetry module
Button Description

sample]
[Measure If you have not saved any sample sequences to the method, the Sequenz
sequence] einstellen window will appear after clicking on this button. This window
may only be exited once there has been at least one sample defined.
Measure a reference
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Colorimetry module

measurement.
Execution
with samples.

Button Description
Stop measurement
processed further.
[Sequence The current measurement step is stopped and then the processing
suspend] interrupted. The measurement pause may be used to view and edit the

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Colorimetry module
[Measure sequence].
switch to the Measurement worksheet.
10.3 Displaying, analyzing and editing results in the Colorimetry module
This section explains the particularities of the results display in the Farbmetrik module
window. Please read up on the general structure of the document windows in section
"The document window" p. 15.
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Colorimetry module
Document window in Farbmetrik module
1 Measurement worksheet 3 Measured values spreadsheet

2 Color view worksheet 4 Color values spreadsheet
Measurement worksheet The Measurement worksheet includes the original data. The sample spectra are
displayed in the upper half of the worksheet.
Sample table The Measured values spreadsheet contains the sample table in the order of the
measurements and the table with the digital measured values of the sample spectra.
The sample table can be configured freely. After right clicking on the table header the
following parameters can be selected in the context menu:
 Color of the spectrum in the graphic
 sample name
 Notes
 Sample type
 Temperature for Peltier-tempered accessory
The individual measurements for each sample are listed in a separate table.
Output of color values On the Color values spreadsheet, the data analyses already defined in the method are
shown. These analyses can be amended and edited.
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Colorimetry module
Color view worksheet The Color view worksheet displays the chromaticity coordinates and their position in the
coordinate system:
Option/value Description
Parameter Variation of parameters for calculating the chromaticity coordinates:

Observer Select viewing angle 2° or 10°.
Light type Standard light type A, C or D65.
Note:
The selection on this spreadsheet does not have an effect on the results
output on the Color values spreadsheet. The pathlength is set at 1 cm,
regardless of the pathlength agreed.
Wavelength Dominant wavelength of the hue on the standard color value chart
Saturation On the standard color value table, the saturation of the hue is the ratio of the
distance of illuminant (achromatic point)/color coordinates (x, y) and
illuminant/dominant wavelengths on the standard color value chart.
Coordinates xy values of the hue on the standard color value chart.
Display of color values with color value chart
10.3.1 Display options for the spectra in the Colorimetry module
The following functions are available for displaying the color spectrum:
 Scaling curves
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Colorimetry module
10.3.2 Analyzing spectra in the Colorimetry module
The data analyses offered in this method can also be performed later.
Display of the analysis The analyses are shown in the lower part of the worksheet on the Color values
spreadsheet.
Color values spreadsheet
Edit analysis The data analysis parameters defined in the method (→ "Colorimetry – Settings / " p.
128) can be edited.
Execution
 In the Color values spreadsheet, click on Settings.
 In the Settings – Color values dialog window, activate the desired color coordinates
with the parameters or the color numbers to be calculated.
 The view on the Color values spreadsheet is adjusted depending on the
selected options and parameters.
10.4 Exemplary measurement in the Colorimetry module
As an example, the tristimulus values under the viewing angles of 2° and 10° with
standard light type A are determined. For the measurements, an integrating sphere and
monochrome paper can be used. If unavailable, a cell with water can be used as
reference and a color solution as sample. See operating instructions " Accessory
SPECORD PLUS" for information on how to handle the integrating sphere.
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Colorimetry module
Open the method and

create a document Click on [Colorimetry] in the launch bar or select Modules  Colorimetry of the
main window.

parameters
 Enter the parameters in the method window.
 Exit the parameter input by clicking on [OK] in the Colorimetry - Settings window
General page:

Color valuesscreen:
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Colorimetry module
Sample sequencescreen:
table:
Automatic save page: Make no entries.

Note page: Enter "Example for color measurement".

measurement
 Insert the reference sample (reflectance standard from integration sphere
accessory) and click [Yes] to start the measurement.
displayed.
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Colorimetry module
 Insert sample 1 (color paper) and click on [Yes] to start the measurement.
 The sample spectrum appears in the document window.
Example of a color measurement
138
General functions of the SPECORD PLUS – Menu Device
11 General functions of the SPECORD PLUS – Menu Device
11.1 Show device status
After selecting Instrument  Info in the menu, the device status of the SPECORD PLUS is
displayed. After successful initialization, display information on the firmware version
and on connected accessories.
If the SPECORD PLUS is not connected to the computer, the message "Simulation mode:
SPECORD XXX PLUS" is shown, whereby XXX corresponds to the device configured in the
window Options / Start / Simulation (Settings  Options).
11.2 Initializing the SPECORD PLUS and starting simulation mode
In accordance with the standard settings in the window Options / Start / Start ASpect
UV, the device is initialized when the ASpect UV program is started and SPECORD PLUS
is switched on, i.e. there is a connection established between the device and the
computer. The SPECORD PLUS is then ready to perform measurements (→ "Defining the
start mode for ASpect UV" p. 154).
The ASpect UV program runs in simulation mode if it is not connected to SPECORD
PLUS. In this mode, existing measured values may be analyzed and processed. In
addition measuring functions are simulated; the program can be explored without the
SPECORD PLUS being switched on.
If the connection to the SPECORD PLUS fails while starting the ASpect UV, ASpect UV
automatically switches to simulation mode. The device will be ready again to perform
measurements following a subsequent device initialization.
Initialize SPECORD PLUS Execution

 If necessary, install and connect the accessories in the SPECORD PLUS. Switch on
the SPECORD PLUS.
 Close all documents in Aspect UV.
 Click on the [Initialization] button on the launch bar or select Instrument 
Initialization.
 The monochromator of the SPECORD PLUS moves and on the monitor the
message "Initialization" is shown. If the options to switch on the lamps are not
activated in the window Options / Start / SPECORD PLUS under While switch
on the device, a prompt relating to switching on the lamps will appear.
After completing the lamp activation phase, any connected accessory will be
initialized and the SPECORD PLUS is then ready for measurements.
Start simulation mode Execution

 Close all documents in Aspect UV
139
 Select Instrument  Simulation from the menu.

 The USB connection to the device is closed and the device and accessory chosen
in the window Options / Simulation mode are activated.
Note: If no further measurements are to take place, the SPECORD PLUS may be
switched off at the power switch.
11.3 Correct SPECORD PLUS
Basic and grating The SPECORD PLUS is fully adjusted and set up ex factory. Changes in the adjustment
correction caused by transport, ambient temperature or change of lamps will be corrected by the
ASpect UV program, without requiring mechanical intervention in the device.
The following parameters are checked and corrected where necessary during the Basic
correction:
 Offset (dark current)
 zero order of the lamps
 amplifier stages of the receivers
 Wavelength correction using internal holmium filter and UV lamp
In the SPECORD 250 PLUS, a grating correction is additionally performed and the
grating of the pre-monochromator aligned with the grating of the main
monochromator.
Attention Only perform the basic correction after 2 hours warm-up time. During this period both
lamps must be switched on.
In the SPECORD 250 PLUS start the grating correction first and then the basic
correction.
Execution
 Remove samples and accessories that influence the beam path (integration sphere,
flow cell etc.) from the beam paths in the sample space.
 Only SPECORD 250 PLUS:
Select Instrument  Correction  Grating correction from the menu.
 The grating correction starts immediately.
 Select Instrument  Correction  Basic correction from the menu.
 The basic correction starts immediately.
 Once the correction is complete, you have the option of showing the results of the
correction.
At the end of the correction process, the new correction data is permanently saved in
SPECORD PLUS and on the computer. The data remain in place until the correction
values are recorded the next time and are used for the correction of all subsequent
measurements.
Note: If accessories requiring elaborate installation, such as Peltier-tempered changers,
are solely used for measurements and they were already installed before initialization,
140
they may remain in the sample space if accessory detection has been switched off before
the correction. Only the samples will need to be removed from these accessories
beforehand.
Correcting infinite Correction of the infinite absorbance is only possible if the “Validation according to USP”
absorbance module has been installed. Further information on infinite absorbance can be found at
"Supplement B - Generation of measured values and recording of reference values" p.
211.
Execution
 Carry out a grating correction (with SPECORD 250 PLUS) and basic correction (see
above).
 Select Instrument  Correction  Correction of infinite absorbance from the menu.
 Recording of the correction spectra starts.
At the end of the correction process, the correction spectra are saved in Aspect UV on
the computer. The data remain in place until the correction spectra are next recorded
and are used for all subsequent measurements.
11.4 Checking the measuring technology condition of the SPECORD PLUS.
ASpect UV provides functions for checking the measuring technology condition of

SPECORD PLUS. This allows for validation of the correctness of the measured values and
makes it easier to find device errors and faults. The functions for checking SPECORD
PLUS are located in the menu Instrument.
11.4.1 Checking the lamp intensity
The halogen lamp for the Vis range and the deuterium lamp for the UV range are subject
to normal wear. With aging, the beam properties deteriorate resulting in a worse signal-
to-noise ratio and thus a lower photometric reproducibility and a reduced photometric
range. The lamp test checks whether the energy of the lamps is still sufficient to meet
the technical specification of the SPECORD PLUS.
Attention If one or both of the lamps have been replaced, the device correction must be carried out
again (→ "Correct SPECORD PLUS" p. 140).
Lamp test Execution

 In the main window, select Instrument  Test  Lamp check.
 The energy measurement of the UV lamp followed by the Vis lamp takes place.
141
After completion of both measurements, the bar charts of the measured results of the
tested lamps and the information about the lamp condition are displayed in the window
Lamp check:
 The lamp energy is within the standard range
 The lamp is nearing the end of its service life
 The lamp should be replaced
11.4.2 Checking the functionality
Scope of the functional The ASpect PLUS program includes a diagnostic program for checking the following
check functions of the SPECORD PLUS:
 Energy in the measurement and reference beam path
 Reference measurement as ratio of measuring and reference beam path in %T
 Blank measurement in %T as 100-percent base line
 Wavelength accuracy through registration of the spectral lines of the deuterium lamp
at 485.9 nm and 656.1 nm
The diagnostic results can be printed and saved in a file.
Functional check Execution

 In the main window, select Instrument  Test  Instrument check.
 Click on [Start].
 The test items are processed automatically.
After completion of the test, the measuring graphs are displayed on the 4 tabs of the
window Instrument check. In case of device errors, corresponding messages are output.
142
Graph display after completed device test
For the graph view, the graphical zoom function is available.

 With the mouse button held down, draw a frame around the area to be enlarged.
The action is reversed by double clicking on the diagram.
The results of the device check may be saved, reopened and printed using the File
menu.
11.5 Measuring energy
During the energy measurement, the energy progression is recorded separately in the
reference and measurement beam paths and output as energy values. This function is
mainly used by Service to evaluate the device condition and find faults.
After selecting Instrument  Test  Energy, the Energy measurement window appears.
Measurement parameter The measurement parameter configuration takes place in the same way as the
configuration configuration in the Spectrum module with the following limitations:
 No measuring mode can be selected. Only the energy signal is recorded
automatically.
 No calibration is provided.
 In the sample sequence, only the sample type Sample is permitted.
See also section "Method configuration in the Spectrum module" p. 71.
143
Start/analyze energy After selecting the measurement parameters, the buttons for starting the energy
measurement measurement are released.
Execution
 Start energy measurement with .
 Add further measurements, as required, with or .

 To analyze the data, the functions Peaks, Values of def. wavelengths and Formula
may be used (→ "Analyzing spectral data" p. 80).
11.6 Setting the zeroth order
With the function Zeroth order, the monochromator is rotated into a position where the
light is not divided at the grating but reflected like in a mirror. The undivided (white)
light passes through the sample space and can be easily monitored due to its intensity.
This is used to adjust accessories in the beam path, e.g. microcells. The zeroth order may
be separately set for the halogen lamp and the deuterium lamp.
Execution
 To configure the zeroth order of the deuterium lamp, select Instrument  Zeroth
order  UV lamp.
 To configure the zeroth order of the halogen lamp, select Instrument  Zeroth order
 Vis lamp.
 The lamp radiation not passes undivided through the monochromator and
becomes visible in the sample space.
11.7 Additional functions for controlling accessories
The following functions for controlling accessories are available in the menu Device:
 Switching active accessory detection on and off
 Switch active accessories on and off electrically
 Move cell changers and samplers to individual positions
 Adjust cell changers
 Optimize pump times
 With a Peltier-tempered accessory, prompt for temperature range and current
temperatures
11.7.1 Switching active accessory detection on and off
The detection of active accessories can be switched off with software support without
this having to be removed from the sample space or the identification plug having to be
pulled.
In the subsequently opened method parameters, the accessory is treated as a passive
accessory, even though the accessory continues to be supplied with power.
144
Execution
 In the main window, select Instrument  Accesory  Accessory detection off.
 The accessory detection is switched off and no longer used for the subsequent
measurements. The accessory continues to be supplied with power.
 To re-activate the accessories, select Instrument  Accessory  Accessory detection
on from the menu.
 The accessories are available again for methods and measurements.
11.7.2 Switching accessories on and off
With this function, the electrically controlled accessory is switched off. The accessory is
also moved to the parking position and the power supply interrupted. The accessory can
then be easily removed from the sample chamber without risk of short circuit.
Execution
 In the main window, select Instrument  Accessory  Off.
 The accessory is switched off and will not be displayed in the method
parameters anymore. The accessory can then be removed from the sample
chamber.
 In order to reactivate the accessory, select Instrument  Accessory  On from the
menu.
 The accessory is initialized and available again for methods and measurements.
Note If the connector is not removed from the accessory, the accessory will be automatically
recognized during the next initialization of the device, unless the accessory detection
has also been switched off.
11.7.3 Adjusting the cell changer and moving to individual positions
Moving to positions in the The cell changer or cell carousel may be moved to a specific sample position.
cell changer or cell
Execution
carousel
 In the main window, select Instrument  Accessory  Sample position.
 In the window Sample position move the cell changer to the desired position with
the buttons:
Button Description
[Pos. 1] Move the cell changer to position 1 and initialize
[Pos. +1] Move the cell changer to the next position
[Park] Move the cell change to parking position

In the parking position it is easier to assemble the cell changer in the sample
space or pack it into its storage box.
 The cell changer moves to the specified position.
145
Window Sample position for moving to a position in the cell changer
Adjust cell changers To position the cells optimally within the beam path the cell changer can be adjusted
with the aid of a computer.
An adjustment is necessary in case of
 the first use of the cell changer
 the use of special cells
 after a basic correction
 after transporting the SPECORD PLUS
Execution
 Assemble the empty cell changer in the sample space.
 Start ASpect PLUS and switch on SPECORD PLUS.
 When using microcells:
Fill the microcells with solvent (e.g. distilled water) and populate every space in the
sample changer with them.
When using standard or semi-microcells:
Empty all spaces in the cell changer.
 Start adjustment with Instrument  Accessory  Adjust.
 The adjustment is carried out automatically and the corrected values saved to
the device and the PC.
11.7.4 Optimizing the pumping time of the cassette sipper system
When calculating the optimum pumping time, there is a measurement taking place
while the sample or reference solution is pumped through the cell. The change of
energy, absorbance and transmission on 3 tabs over time is displayed on the screen.
During the process the measured value increases or decreases and finally reaches a
plateau. The optimum pumping time corresponds to the reaching of the plateau phase.
At this time the cell has been flushed sufficiently with sample or reference and the carry-
over is smallest.
146
Functions of Optimization
of pumping time window
Optimization of pumping time window for the cassette sipper system
In the menu bar the following buttons are available:
Button Description
Menu for data administration
File Save Save optimization data
Open Open optimization data
Close Close Pumpzeit optimieren window.
Start optimization.
Start
Switch pump on/off independent of starting the measurement.
Pump on/off
Stop the measurement early.
Stop
On the tab Configuration the following parameters must be entered:
Wavelength Enter the analysis wavelength of the sample.
Measuring Enter the total measuring time for the optimization.

time
Integration Select the time for recording a measuring point, e.g. 0.1 s.
time
Slit Only SPECORD 210/250 PLUS:
Select the slit setting, e.g. 1 nm.
147
Start pump Time during the measurement from which the pump is running.
Stop pump Time during the measurement at which the pump stops.
Slow run The pump speed is reduced by 50% in order to lower the degassing or
segregation, or to allow for more accurate dosing with smaller sample
volumes.
Energy Enter the ordinate range for the display of the measured values as energy
value.
Absorbance Enter the ordinate range for the display of the measured values as
absorbance value.
Transmittance Enter the ordinate range for the display of the measured values as
transmittance value.
The measured values are displayed graphically during optimization on the tabs Energy,
Absorbance and Transmission.
Perform optimization Execution

 Provide reference and sample solution.
 Start optimization by selecting Instrument  Accessory  Optimize pumping time.
 Remove the flow cells from the holder and fill them with reference solution:
− Submerge the aspiration hose into the reference solution and click on .
− Once the cell is filled without bubbles, stop the pump by clicking on .
 Place the flow cells in the holder and submerge the aspiration hose into the sample
solution.
 Start the measurement by clicking on .

 To start, the reference is automatically determined and then the measured
values are recorded.
 Observe the change of the measured value on either one of the tabs Energy,
Absorbance or Transmission:
− The time range during which the pump is running is shown on a white
background. The range during which the pump was stopped is gray.
− The pump time is optimal if the measured value is stable or has reached the
required accuracy.
− During the time after stopping the pump it can be observed whether the sample
remains stable or still changes due to decreasing turbulence.
11.7.5 Adjusting the sampler, moving to individual positions and optimizing the pumping
time
Moving to individual Individual positions of the sampler can be moved to separately.

positions
Execution
 In the main window, select Instrument  Accessory  Sample position.
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 In the window APG xyz sampler, move the sampler to the desired position with the
buttons:
APG-116 xyz-Probengeber window
Adjust the autosampler Depending on the size of the opening on the sample containers, the sample positions
may need to be readjusted.
Execution
 Place a sample container at the 4 corners of the sample tray
 Lower the cannula holder without cannula using the arrow (downwards) in the
APG- xyz-Probengeber window. Then insert the cannula and position slightly over
the edge of the container.
 Centrally position the cannula over Off X and Off Y. Repeat the process at the three
other corners and determine the values for Off X and Off Y between the 4 corners.
Off x / Off y Adjust the sample aspiration hose with the center of the container in
the X and Y direction
Adjust the Z-lift of the sampler arm
[factory settings] Restore the factory settings
Pos. 0 Initialize APG and move to position 0

Note: A move to the start position is only possible in the positioning
window. The system can be pre-rinsed with a separate purge cup.
Tip
The final immersion depth is set by moving the sample aspiration hose with the sample
arm lowered in its guide (→ "Operating instructions SPECORD PLUS Accessories").
APG S sampler For the optionally available APG S sampler, ASpectUV additionally provides the option of
adjusting the sample tray to meet customer requirements.
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APG-S xyz Sampler window
Parameter / Description
button
nx / ny Set the number of sample containers via the number of table rows (nx)
and columns (ny).
nx0 / ny0 Shift position 1 in x/y direction
lx / ly Set the sample position via the spacing of the sample tubes relative to
each other.
Note: The graphic representation in ASpect UV is not adjusted.
column by Swap the direction of the numbering of the sample locations.

column
Optimize pump time The pump time of the cassette sipper system must be optimized for the transport path
of the sampler.
 Provide reference and sample solution.
 Start adjustment by selecting Instrument  Accessory  Optimize pump time.
 Click on Sample position and move to the position of the reference.
 Lower the autosampler arm with Needle down.

Continue as described for the pump time optimization of the cassette sipper system (→
"Optimizing the pumping time of the cassette sipper system" p. 146). Move to the
positions on the APG with the buttons , and and lower or raise the sampler
arm.
150
Window Optimization of pumping time for APG
151
Setting ASpect UV in the Options window
12 Setting ASpect UV in the Options window

A number of functions for setting the view, the start and the execution of the program
are included in ASpect UV. For example, a quick start menu with your most used
methods can be displayed at program start or the calibration in the individual modules
can be preconfigured. These settings are made in the Options window.
Options window
The pages of the Options window are sorted by topic for better overview.
Group Content
General Configuration of the launch bar, quickstart, directory path presets

and language
Start Settings for program start, the device and the simulation mode
Measurement / Settings for reference measurement, calibration and decimal points

display
Advanced Configuration for signatures, passwords, archiving, CSV export and

webservers
 The Options window is opened via menu item Settings  Options.
12.1 Configuring the launch bar
The launch bar provides quick access to the individual modules and functions of ASpect
UV. The launch bar is configured in the Options / General / Launch bar window.
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Options / General / Launch bar window
Procedure
 Open the Options window via menu item Settings  Options.
 On the General / Launch bar page, activate the checkboxes for the desired
elements.
 Drag&drop (with the mouse button held down) the active elements to sort them in
the desired order.
 After exiting the Options window with [OK], the elements are displayed in this
order in the launch bar.
12.2 Configuring quick start
Quick start is a collection of methods in which routine methods may be saved. When
quick start is accessed, these methods are available for use and may be started
immediately. Configure the quick start menu in the Options / General / Quick start
window.
Procedure
 On page General / Quick start, click on .

 Enter the name to list the method in the quick start menu in the input field.
 Click on and select the method.

 Any number of methods can be stored in quick start in this manner.
A method that is not required can be removed from the quick start as follows:
 Highlight the method that is not required in the list on the General / Quick start
page.
 Click on and confirm deletion of the element.
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 The element is removed from the quick start.
12.3 Directory path presets
Directories for loading and saving data (methods, results, calibrations, print and export
templates, sample data) can be preset in the Options / General / Paths window. This
path can be changed when loading or saving the data.
Procedure
 Select the module to preset directory paths for in the Module list on the General /
Paths page.
Select All modules to set the same directory paths for all modules.
 Select a function in the list, for example Open method, and select a directory option
in the list next to it:
− Last used path: The last path used will be used the next time the selected data is
loaded/saved.
− Default path: The path set here is displayed at next load/save.
 Click on […] and set the path in the Select directory window.
Note: Open a file via menu items File  Open results or File  Open method, the
directory for the last module opened is then displayed.
12.4 Setting the language
Change the program language in the Options / General / Setting language window.
Procedure
 Select the language on the General / Setting language page.
 The new language is immediately displayed in the main window and in newly
opened documents.
Documents that are already open will partly contain the previous language. Close and
open these documents again in order to show these documents with the current
language setting.
12.5 Defining the start mode for ASpect UV
The start options for ASpect UV are set in the Options / Start / Start ASpect UV
window. Automatic initialization and the program element to display when the device
starts can be set.
Procedure
154
 Set the following on the Start / Start ASpect UV page:
Option Description
Initialize When starting ASpect UV, connect SPECORD PLUS with the computer and
instrument initialize.
Note:
First switch on SPECORD PLUS and wait for the internal device initialization
to complete. Then start ASpect UV.
Open element Select a start element.
Either the modules or the quick start may be selected here (→ "Configuring
quick start" p. 153). If the option None is selected, the main window of
ASpect UV is displayed after starting.
12.6 Switching on the lamps and recording monitor files
Automatic activation of the lamps when the SPECORD PLUS is switched on can be set in
the Options / Start / SPECORD PLUS window.
Procedure
 Set the following on the Start / SPECORD PLUS page:
Option Description
Switch on the When the device is switched on, switch on the UV lamp
UV lamp
Switch on the When the device is switched on, switch on the Vis lamp
Vis lamp
Monitor file If activated, the data traffic between the SPECORD PLUS and the computer is
recorded. This service may be used by Analytik Jena Service for remote
maintenance of the SPECORD PLUS.
UV off Switch off the UV lamp
Vis off Switch off the Vis lamp
If automatic activation is deactivated, a dialog will open asking whether to switch on the
lamps during initialization and start of a method. The lamps are only switched on when
this is answered with [YES].
12.7 Configuring simulation mode
The type of the SPECORD PLUS and the active accessories used for method parameter
input and simulated measurements are set in the Options / Start/ Simulation mode
window.
Procedure
155
 Set the following on the Start / Simulation page:
Option Description
Instrument Select the SPECORD PLUS type.

Different measurement ranges and columns are available in the measurement
parameters depending on the type.
Accessory Select accessories for simulated measurements.
If you wish to simulate non-electrical accessories in the sample chamber, for
example the standard cell holder or the holder for solid samples, select the
Passive accessory option.
12.8 Displaying reference measurements
By default, the reference measurement values are not displayed. They can, however, be
displayed for service purposes or to check the baseline. Select this setting in the Options
/ Measurement /display / Reference measurement window.
Procedure
 Set the following on the Measurement / Display / Reference measurement page.
Option Description
Display measurement Display reference values or graphs during recording.

values
Close automatically after The reference value window is closed automatically after
the measurement recording. Otherwise, there will be a prompt to close the
window.
12.9 Configuring calibration
In the Options / Measurement / Display / Calibration window, the validity of a

calibration graph can be defined and default settings for the calibration in the method
parameters can be changed.
Procedure
 Set the following on the Measurement / Display / Calibration page.
Option Description
R2 adjust A determination coefficient of the R2 adjust regression for recorded

calibration graphs can be entered. Only calibration graphs that meet this
criterion can be used for an analysis.
Period of If activated, a validity period must be entered for the calibration graph. After
validity expiry of this period, new calibration must take place.
Regression Preconfigure the regression function for the method parameters.
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Unit Preconfigure the concentration unit.
Pathlength Preconfigure the pathlength of the cell.
Limit value Preconfigure the limit value for the concentration analysis.
12.10 Defining decimal places
The numbers behind the decimal points displayed and used for export can be defined for
all numerical values in ASpect UV in the Options / Measurement / Display / Decimals
window. These are broken down into General for measured values, and Calibration for
quantitative analyses and for the special analyses in the Kinetics, Colometry and
Thermometry modules. Increase or decrease the number of decimal places with the
arrow keys next to the measuring units.
Procedure
 Make settings on the Measurement / Display / Decimals page.
Note:
The internal calculation accuracy for floating-point numbers is double. When exporting,
make sure the necessary number of decimal places is set in order to reduce calculation
inaccuracies.
12.11 Configuring password properties
If the additional ASpect UV FDA 21 CFR Part 11 module with user management is used,
the criteria for valid passwords can be set in the Options / Advanced / Password
management window.
Procedure
 Set the following on the Advanced / Password management page:
Criterion Description
Minimum Minimum number of characters for the password

length
Upper and The password must contain upper and lower case letters.
lower case
Special The password must contain at least one special character.

characters
Digits The password must contain at least one digit.
Number When a new password is created, the system checks whether the user has
used it before. If a password has already been used (Number), it will be
rejected.
Further information on the use of passwords can be found here (→ "Passwords in

ASpect UV" p. 201).
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12.12 Creating signatures
If the additional ASpect UV FDA 21 CFR Part 11 module is used, Aspect UV files can be
signed. A signature limits further editing options for a document. The limits can be
freely selected for every created signature. Optionally, the signature sequence to apply
to a document can be defined.
In the Options / Advanced / Signatures window, signatures can be created, the
properties of the signatures can be configured, and the signature sequence can be
defined.
On the Signatures page, existing signatures with their stored functions are displayed in
the List of signatures tab, and signatures are created and deleted. A required signature
sequence can be defined in the Sequence of signatures tab.
Adding, editing and Signatures are managed in the Options / Advanced / Signatures n window in the List
deleting signatures of signatures tab.
Procedure
 Switch to the List of signatures tab on the Advanced / Signatures page.
 To add a signature, click on to open the Append signatures window.

− Enter the signature in the Name input field and an additional description in the
Description field.
− In the Document, Measurement procedure and Worksheet tabs, activate the
functions that can still be carried out after the signature is applied.
 To edit an existing signature, double-click on the corresponding signature in the list.
 Mark a signature that is no longer required in the list and click to delete it. Only
signatures not included in the signature sequence can be deleted.
Attention Functions that can no longer be carried out after application of a signature will remain
disabled after the application of further signatures.
Defining a signature If a signature sequence is defined, signatures included in the sequence can only be
sequence applied in the indicated sequence. A document is considered fully signed when all
signatures have been applied. For incompletely signed documents, a message that the
file has not been completely signed appears when saving and opening the file.
Signatures in the Available signatures list are not included in the signature sequence.
They can be used on a document at any time and in any sequence.
Procedure
 Switch to the Sequence of signatures tab on the Advanced / Signatures page.
 Select a signature in the Available signatures list and transfer it to the The order to
be following when signing list with the arrow key.
 Transfer all further required signatures to the The order to be followed when
signing list in the same manner.
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 Sort the collected signatures in a sequence. Mark each signature and move it to its
required position with the arrow keys to the right of the list. Work downward from
the top of the signatures list to the bottom
 Mark the signatures not required in the sequence and transfer them back to the
Available signatures list with the arrow key in the middle.
Note: Permission to use a specific signature must be assigned to each user in their user
profile (→ "Create and edit user profiles" p. 197).
12.13 Activating automatic archiving
Automatic archiving of documents can be set in ASpect UV. To do this program-wide,

activate the Automatic archiving option in the window and select the time for each
archiving function. Define the file name and the report options for automatic archiving
in the method (→ "Automatically archiving documents" p. 30).
Options / Advanced / Automatic archiving window
Procedure
 To activate automatic archiving program-wide, activate the Automatic archiving
option on the Advanced / Automatic archiving page.
If this option is deactivated, Automatic archiving is deactivated program-wide,
regardless of whether this function is activated in a method.
 Select a function, for example Save, in the Archiving function list.
 Activate the options to carry out the selected archiving function, e.g., When closing
the document.
 Define all further archiving functions in the same manner.
159
12.14 Configuring the CSV export
Settings for the manual CSV file export from a document window and for the automatic
CSV file export defined in the method parameters can be made in the Options /
Advanced / Settings CSV Export window.
Procedure
 Set the following on the Advanced / Settings CSV export page:
Option Description
Transposed When activated, columns and rows in the CSV export file are transposed.
For example, in the Spectrum module, the default is that the wavelength is
written continuously in one row, and the corresponding measured values in
the next row. In the transposed display, output is by column: one column with
the wavelengths, and the corresponding measured values in the next.
Column The following column separators can be selected:
separator  Space
 Semicolon
 Comma
 Tabulator
12.15 Configuring the webserver
Configure the network for the use of the web server. These settings must be made by a
system administrator (→ "WebApp" p. 204).
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Validating the SPECORD PLUS (optional)
13 Validating the SPECORD PLUS (optional)

For quality assurance when working with the SPECORD PLUS, performing a validation at
regular intervals is recommended. The following modules are optionally available for the
various requirements in the individual labs:
 Validation AJ: comprehensive validation of the measuring technology properties in
accordance with the technical data of the manufacturer
 Validation Standard Ph. Eur.: Validation according to the requirements of the
European Pharmacopeia
 Validation USP: Validation in accordance with the requirements of the
Pharmacopeia of the United States of America
 Validation Maintenance economic: Validation as part of a service agreement
13.1 Validation AJ
Validation AJ is a comprehensive validation of the measuring technology properties in

accordance with the technical data of the manufacturer (see Technical Data in the
operating manual "SPECORD PLUS UV/Vis spectrophotometer). Furthermore, personal
limit values may be determined for the validation in order to validate the device for a
specific measurement task.
Validation module The Validation regarding AJ module window appears after selection of Validation 
window Test  Validation  AJ.
Validation regarding AJ window
The scope of the parameters to be verified is determined under [Setup]. The

validation results are displayed on a separate tab for each test parameter. The Results
161
tab outputs the digital test results in each case and the Measurement values tab a
diagram of the measured spectra. In the Part of validation list field, a parameter group
can be selected for the display in each case.
The audit trail may be shown on an additional tab. The results of the validation may be
saved, exported, printed and opened again.
Attention To achieve optimum results, observe the following information:

 Start the validation after a device warm-up period of 2 hours. During this period,
both lamps of the SPECORD PLUS must be switched on.
 Perform a basic correction directly before opening the Validation window or, for the
SPECORD 250 PLUS, a grating and basic correction.
13.1.1 Test parameters and required equipment
Test parameters The following parameters can be checked using the device validation:
Transmittance zero Measurement with covered measurement beam path and open
point reference beam path in the range of 200 to 1000 nm
The minimum and maximum of the zero point transmittance of the
device are calculated from the result.
Baseline deviation Recording of the baseline in the absorbance mode in the range of
200 to 510 nm
Baseline noise Recording of 30 measuring cycles of the baseline in the range 490
to 1000 nm
Determination of the RMS noise (standard deviation) at 500 nm
100% transmittance Measurement of the baseline in transmittance without correction

uncorrected Determining the minimum and maximum transmission
Photometric accuracy Test with a certified standard filter
Vis range Determination of difference between measured absorbances and
specified target values of the standard filter set at 5 wavelengths
Photometric accuracy For SPECORD 210 PLUS with extended measuring range
in the Vis and NIR Test with a certified standard filter
range Determination of difference between measured absorbances and
specified target values of the standard filter set at 10 wavelengths
Photometric accuracy Measurement of the absorbance of a potassium dichromate solution
UV range at 4 wavelengths in the range of 235 to 350 nm
Determination of difference between measured absorbances and
specified target values of potassium dichromate solutions
Photometric accuracy Measurement of the absorbance of a potassium dichromate solution
at 430 nm at wavelength 430 nm
Determination of difference between measured absorbance and
specified target value of potassium dichromate solutions
162
Wavelength accuracy Measurement of the peak wavelength positions of a holmium glass

filter or a holmium oxide solution with known target values for the
wavelengths in the five ranges 270 to 285 nm, 350 to 370 nm,
450 to 485 nm, 530 to 545 nm and 630 to 645 nm.
Wavelength accuracy Measurement of the wavelength position of the D2E lines (485.99
at 486 nm and 656 nm and 656.1 nm) at the wavelengths 486 nm and 656.1 nm
nm/UV lamp
Wavelength Performance of ten measurement cycles at 358 - 365 nm for
reproducibility determining the standard deviation of the wavelength position of
the holmium oxide peak
Stray light Determination of the maximum value of stray light in the

transmittance mode with different cut-off filters in the following
ranges
Cut-off filter Measuring range
KCl filter cell 198 nm
NaJ filter cell 220 nm to 240 nm
NaNO2 filter cell 340 nm
Spectral resolution Test of the spectral resolution capability with a toluene solution in n-
hexane by determining the absorbance ratio of the wavelength 269
nm to the wavelength 266 nm (A269/A266)
Longterm stability Repeated measurement of the baseline over an hour and calculation
of the gradient of the measured values at 500 nm
Test equipment For the validation of the SPECORD PLUS, the following filters and certified standards are
required:
 Certified filter kit for the photometric measurement of the Vis spectral range and
certified holmium oxide glass filter for the wavelength accuracy
 Standard solution of holmium oxide in perchloric acid for checking wavelength
accuracy (alternative to the holmium oxide glass filter)
 Potassium dichromate standard solution for the UV photometry and for 430 nm
 Sodium nitrite standard solution for the stray light measurement
 Sodium iodide standard solution for the stray light measurement
 Potassium chloride standard solution for the stray light measurement
 Referenced water cell or bidistilled water (free from gas bubbles)
 0.02% standard solution toluene in hexane and hexane as reference for spectral
resolution test
Attention For the certified standards, the manufacturers specify a temperature range for use on
the certificates. You must comply with this temperature range during validation. A
validation outside the specified temperature tolerances can result in incorrect
measurement results!
163
13.1.2 Entering validation parameters
At the start of the validation, the parameters to be validated need to be selected.

Execution
 In the main window, select Instrument  Test  Validation ▸ AJ from the menu.
 Click on [Setup].
 In the Settings window, select the following parameters:
Tab Description
General Name of the operator

If the user management is is installed, the logged on in the software user is
displayed.
Selection of the parameters to be tested
Acceptance Configuration of the limit values and tolerances of the parameters to be
criteria tested
all other tabs Test equipment parameters (designation, test wavelength, target values,
etc.)
The entries on the respective tab must be made if the corresponding test
parameter has been selected on the tab General.
 Confirm the configurations of the parameter limits and values with [OK].
 The Configuration window is closed and the button (Start) released in the
Validation window.
Select test parameters –

General tab
Select validation test parameters, Settings / General window
The parameters to be tested can be freely selected to allow your own validation routine
to be defined for the spectrometer.
164
Test parameters Description
General Transmittance zero point; baseline deviation; baseline noise; 100% T

line, uncorrected
Photometric Photometric accuracy in the Vis range

accuracy Vis
Photometric Only for SPECORD 210 PLUS with extended measuring range
accuracy Vis / NIR Photometric accuracy in the Vis range and in NIR range (up to 1200
nm)
Photometric Photometric accuracy in the range 235 – 350 and at the wavelength
accuracy UV / 430 430 nm
nm
Wavelengths Wavelength accuracy and wavelength reproducibility in the range 250
nm to 650 nm
Stray light Stray light at 198 nm, 220 nm, 240 nm and 340 nm
Spectral resolution Spectral resolution capability with a toluene solution in n-hexane by

determining the absorbance ratio of the wavelength 269 nm to the
wavelength 266 nm (A269/A266)
Long-term stability Measurement of the baseline over an hour and calculation of the
gradient of the measured values at 500 nm
Select limit values – The limit values for the device test can be set in accordance with the parameter limits
Acceptance criteria tab and tolerances guaranteed by the manufacturer or within your own defined range limits.
In the latter case it will be ensured that the spectrometer is suited for a specific
measuring task.
Entering limit values for the validation, Setting / Limit values window (for SPECORD 250 PLUS)
165
Option Description
Default Use parameter limits and tolerances guaranteed by the manufacturer

values AJ The data corresponding to the technical specification is displayed in the input
fields of the table. They cannot be edited.
Values Use own data record with parameter limits and tolerances
from file New Enter new limit values.
Save Save limit values. The files are given the extension
"*.vat" (validation tolerances)
Open Load saved limit values and use for the validation
Enter key data of the

Enter the identification numbers and target values of the resources used on the
standards used
respective tabs if you have selected the corresponding test on the GENERAL tab.
Photometric accuracy Vis tab
Name / Id. Identification number of the certified standard filter kit (gray filter) in
accordance with the datasheet
Filter 5 test wavelengths and related filter absorbances by absorbance value 1
parameters
Photometric accuracy Vis/NIR tab

Only for SPECORD 210 PLUS with extended measuring range
Name / ID Identification number of the certified standard filter kit (gray filter) in
accordance with the datasheet
Filter 10 test wavelengths and related filter absorbances by absorbance value 1
parameters
Photometric accuracy UV / 430 nm tab
Reference Identification number of the reference (0.01 N sulfuric acid

or perchloric acid)
Potassium dichromate Enter the identification number of the potassium dichromate
solutions with the concentration 60 mg/l and 600 mg/l.
Test wavelengths and Enter wavelengths and target values for the test in
nominal absorbance values accordance with the datasheet.
Wavelengths tab
Name / Id. Identification number of the wavelength standard used:

Holmium oxide filters or holmium perchlorate standard solution
Test wavelengths Target values of the spectral peak in accordance with the datasheet
Stray light tab
Reference Identification number of the reference
KCl Identification number of the potassium chloride standard solution
NaI Identification number of the sodium iodide standard solution
166
NaNO2 Identification number of the sodium nitrite standard solution
Spectral resolution tab
Reference (Hexan) Identification number of the hexane reference solution
Toluene/Hexane Identification number of the toluene standard solution
13.1.3 Running the validation
Attention Before starting the validation the SPECORD PLUS must have already been switched on
for 2 hours with both lamps. Directly before opening the window Validation regarding
AJ, a correction of the SPECORD PLUS must be performed (→ "Correct SPECORD PLUS" p.
140).
Place filters and cells All filters and cells with standard solutions used, including references, must always be
placed in the measurement beam path. The reference beam path remains empty for all
measurements!
Running the validation After entering the validation parameters in the window Settings, the button [Start]
is released. The validation runs assisted by the program. Dependent on your selection of
test parameters in the Settings window, the measurements are performed
consecutively.
 Start the validation by clicking on

 Follow the subsequent instructions for the use of filters and cells in the windows.
Window with instruction to insert a filter
 After each partial validation step the test results and defined tolerances are
displayed in a window. With its buttons the test results window offers the following
options for the subsequent validation method:
Option Description
Repeat Repetition of the test item.

The measurements already recorded for the current test item are discarded.
Note: The repetition of a test item is documented in the audit trail.
Continue Start the measurement of the next test parameter.
Abort Cancel the validation. However, measurements taken so far are temporarily saved
and displayed on the tabs in the main window.
167
Display validation results After completing the validation, the results are displayed on the tabs of the Validierung
window.
Validation regarding AJ window with results
In the list Part of validation, the group of the test parameters to be displayed is
selected. The Results tab displays the digital measurement results in comparison to the
permissible tolerances. The spectra are shown on the Measured values tab, whereby
each spectrum is shown on its own tab. Select File ▸ Overview to display a concise
overview of the validation results.

 In the document window, select File ▸ View Audit Trail from the menu.
 Click on the Audit Trail tab.
 Regardless of the validation part selected, the entire audit trail will be shown on
the far right tab.
Note: The audit trail is also printed with the "Detailed.lst" template.
Export validation results Execution

 In the document window, select File ▸ Export.
 In the ASCII-Export window, enter the name of the export file and the decimal
separator and confirm with [OK].
168
 The data is exported
Print validation results Execution

 In the Open window in the folder \\Users\Public\Documents\Analytik Jena\ASpect
UV\Reports\Validation\AJ, select the "Compact.lst" or "Detailed.lst" print template.
 In the LLPrint Preview window, select the option Print all pages.
 The protocol is printed.
Attention With an operational qualification, the printout is included in the OQ report.
13.2 Validation Standard Ph.Eur.
Validation Standard Ph.Eur. is a validation of the measuring technology properties in

accordance with the requirements of the European Pharmacopeia chapter 2.2.25.
Validation Standard Ph.Eur. comprises all common quality standards. The validation of
the spectrometer is carried out according Ph. Eur. issue 10. Alternative certified
reference materials that have been approved are not available for selection.
Changes as of ASpect UV Ph.Eur. issue 10 requires the validation of the spectrometer in the range of the expected
1.4.2 measured values. The wavelength accuracy should be checked at least 2 wavelengths.
The photometric accuracy (absorbance) should be checked at least at the upper and
lower limits of the expected measurement range. Furthermore, a test of the photometric
linearity was newly included.
In accordance with the requirements of Ph. Eur., the following changes were made in
ASpect UV version 1.4.2 and higher:
 When testing the wavelength accuracy, not only holmium oxide but also dydimium
oxide and the rare earth metal oxide filter for the UV as CRM (certified reference
materials) can now be selected.
 The test for determining the photometric linearity has been newly recorded.
Validation regarding Validation regarding Standard Ph. Eur. module window appears after selecting
Standard Ph. Eur. module Instrument  Test  Validation  Standard Pharm. Eur.
window
169
Window for the validation according to Standard Ph. Eur.
The scope of the parameters to be verified is determined under [Setup]. The

validation results are displayed on a separate tab for each test parameter. The Results
tab outputs the digital test results in each case and the Measurement tab a diagram of
the measured spectra. In the list Part of validation, a parameter group for the display
can be selected in each case.
The audit trail may be shown in a separate window. The results of the validation may be
saved, printed and opened again.
Attention To achieve optimum results observe the following notes:

 Start the validation after a device warm-up period of 2 hours. During this period both
lamps of the SPECORD PLUS must be switched on.
 Perform a basic correction directly before opening the window Validation or, for the
Test parameters The following parameters are checked using the device validation:
Wavelength Measurement of up to 5 standard wavelengths optionally with the rare

accuracy earth metal oxide filters holmium oxide, Didymium glass filter and/or lower
UV
Absorbance Testing of absorbance with potassium dichromate solutions 60mg/l with
UV/Vis wavelengths 235, 257, 313, 350 nm and 600 mg/L at 430 nm
Photometric Test of the photometric linearity at a standard wavelength with 3 or 4
linearity different absorbances or concentrations optionally with neutral glass filter,
potassium dichromate or niacin.
170
Stray light at Stray light measurement at 198 nm with potassium chloride solution 12 g/l
198 nm
Spectral Test of the spectral resolution capability with a toluene solution in n-

resolution hexane by determining the absorbance ratio of the wavelength 269 nm to
the wavelength 266 nm (A269/A266)
This test is optional and only required for the analysis of selected
substances.
Test equipment For the validation of the SPECORD PLUS, the following filters and certified standards are
required:
 Standard glass filter of holmium oxide in perchloric acid or holmium glass filter,
didymium glass filter and/or rare earth metal oxide filters of lower UV for checking
the wavelength in accordance with Ph. Eur.
 Potassium dichromate standard solutions for checking the absorbance in UV and for
430 nm according to Ph. Eur.
 Potassium chloride standard solutions for the stray light measurement according to
Ph.Eur.
 Referenced water cell or bidistilled water (free from gas bubbles)
 0.02 % standard solution toluene in hexane and hexane as reference for the spectral
resolution test
 CRMs with 3 or 4 different absorbances/concentrations at a standard wavelength
optionally with the CRMs neutral glass filter, potassium dichromate or niacin for
testing the photometric linearity
At the start of the validation, the parameters need to be selected.

Execution
 In the main window, select Instrument  Test  Validation  Standard Ph. Eur..
Tab Description
General Selection of parameters to be verified
Acceptance Acceptance criteria of the parameters to be tested according to Ph. Eur.

criteria
All other tabs Test equipment parameters (designation, test wavelengths, target values
and, where applicable, slit size)
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Validation window.
Select test parameters –

General tab
Select validation test parameters, Settings / General window
The parameters to be checked are defined in the European Pharmacopeia. Activate the
parameters you want to check.
View acceptance criteria – The Acceptance criteria tab contains the acceptance criteria in accordance with the
Acceptance criteria tab European Pharmacopeia.
Settings / Acceptance criteria window

Enter the identification numbers and target values of the resources used on the
standards used
respective tabs.
172
Wavelengths tab
Rare earth metall oxide Identification number of the wavelength standard used:
glass filter holmium oxid glass filter, didymium glass filter and/or the
standard for lower UV
Test wavelengths Nominal values of the spectral peak in accordance with the
datasheet
Adsorbanz UV/Vis tab
Reference Identification number of the reference (0.01 N sulfuric acid

or perchloric acid)
Potassium dichromate Enter the identification number of the potassium dichromate
solutions with the concentrations 60 mg/l and 600 mg/l.
Test wavelengths and Enter wavelengths and nominal values for the test in
minimal absorbance values accordance with the datasheet.
Liniearity tab
Type of CRM Select the CRM: Neutral glass filter, potassium dichromate or niacin
Wavelength Test wavelength according datasheet
CRM 1 to CRM Enable the number of used CRM (at least 3)

4 Enter the identification number in the first filed and the absorbance value
in the second filed
Stray light tab
KCl Identification number of the potassium chloride standard solution
Spectral resolution tab
Reference Identification number of the hexane reference solution

(Hexan)
Toluene/hexane Identification number of the toluene standard solution
Slit For SPECORD 210 PLUS and SPECORD 250 PLUS:

Select the slit with (spectral bandwidth). The spectral bandwidth selected
influences the spectral resolution capacity of the device. An overview of
the spectral width and the values to be met can also be found on this tab.
measurements!
173
consecutively.
 Start the validation by clicking on .

Window with instruction to insert a cell
 After each partial validation step, the test results and tolerances in accordance with
the European Pharmacopeia are displayed in a window. With its buttons, the test
results window offers the following options for the subsequent validation method:
Option Description

Interim result during ongoing test
174
Display validation results After completing the validation, the results are displayed on the tabs of the Validation
regarding Standard Ph. Eur. window
Displaying results in the Validation Ph.Eur.
permissible tolerances. The spectra are shown on the Measurement values tab,
whereby each spectrum is shown on its own tab. Select File ▸ Overview to display a
concise overview of the validation results.

 In the document window, select File ▸ Show Audit Trail from the menu.
the far right tab.

175
 The data is exported to the txt-format.

 In the Öffnen window, select the Compact.lst or Detailed.lst print template in the
folder \\Users\Public\Documents\Analytik Jena\ASpect
UV\Reports\Validation\Standard Ph Eur.
 In the LLPrint Preview window, select the option Print all.
Note With an operational qualification (OQ), the printout is included in the OQ report.
13.3 Validation according to USP
Validation according to USP is a validation of the measuring technology properties in

accordance with the acceptance criteria of the U.S. Pharmacopeia <857>.
Note In Aspect UV the version of USP can be selected for validation. Starting with ASpect UV
1.5 the parameters of USP version USP-NF 2021 are also available.
The Validations regarding USP module window appears after selection of Instrument 
Test  Validation  USp.
Validation regarding USP window
The scope of the parameters to be verified is determined under [Setup]. The results
of the validation are shown for each piece of test equipment on two separate tabs with
the name of the test equipment. The top tab includes a selection of the measuring range
176
for the test equipment and the graphical representation of the spectra measured. The
bottom tab shows the digital test results for all test equipment.
The audit trail may be shown on an additional tab. The results of the validation may be
saved, exported, printed and opened again.
Note To achieve optimum results observe the following notes:
Important notes on  Start the validation after a device warm-up period of 2 hours. During this period both
validation lamps of the SPECORD PLUS must be switched on.
SPECORD 250 PLUS, a grating and basic correction, as well as a correction of the
infinite absorbance (→ “Supplement B - Generation of measured values and
recording of reference values” p. 211).
13.3.1 Test parameters, acceptance criteria and required test equipment
Validation according to USP includes the following parameters:

 Accuracy and precision of the wavelength
 Accuracy and precision of the absorbance
 From USP 42: Test of photometric linearity
 Compliance with limit values for stray light
 Checking the resolution
USP involves validation of the spectrometer in the operating range, i.e. the range in
which the expected measured values are found.
In order to cover an extended measuring range both in the wavelength and the
absorbance, a larger number of methods and certified reference materials (CRM,
certified reference materials) is suggested. It is the responsibility of the user to select the
CRM and methods suited to his operating range. The USP also permits, in addition to
the standards it recommends, alternative CRMs once their certification can be
guaranteed.
The test equipment and acceptance criteria for the test parameters are compiled below.
The test parameters apply unchanged for the USP versions 38, 40 and 41. The changes
made in version 42 are marked accordingly in this document. The structuring of the
validation as well as the acceptance criteria of the version USP42, to USP43, to USP-
NF2021 do not differ.
Wavelength In the USP, proof of accuracy and precision of the wavelength is required.
Test equipment / CRM Accuracy Precision
 Hg and D2 emission lines Minimum of 6 measured Minimum of 6 measured

 Rare-earth metal oxide values values
solution (CRM) Difference between average Standard deviation from
 Rare-earth metal oxide value and certified value of average: ≤ 0.5 nm
glass (CRM) CRM:
 Alternative CRM UV: ≤ ±1 nm
Vis: ≤ ±2 nm
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Absorbance Proof of accuracy and precision of absorbance required.

From USP 42: The criteria of photometric linearity are fulfilled if at least at one
wavelength in the UV and/or Vis range with three different absorbances the criteria for
accuracy are fulfilled.
Test equipment / CRM Accuracy Precision
K2CR2O7 solution (235; 257; ≤ ± 1% A above 1.0 A or Minimum of 6 measured

313; 350 nm) ≤ ± 0.010 A below 1.0 A values
Neutral glass filter (440; USP version 38, 40, 41 Standard deviation
465; 546,1; 590 and 635 ≤ ± 0.8% A over 1.0 A or ≤ 0.5% A above 1.0 A
nm) ≤ ± 0.0080 A under 1.0 A ≤ 0.005 A below 1.0 A
From USP version 42:
≤ ± 0.8% A over 1.0 A or
≤ ± 0.008 A under 1.0 A
Alternative CRM
Stray light Filters with a sharply defined spectrum (“cut-off filters) are used for the stray light
measurements, which block any light from passing through below a certain wavelength.
Ideally, filters with a cut-off wavelength that lies as close as possible above the required
wavelength are used.
The USP offers two options for determining the stray light. In the method "10 mm versus
5 mm", a cell with 5 mm path length is first measured as a reference (filled with the
filter solution) and then a 10 mm cell (filled with the same solution) as a sample. The
resulting peak is used to determine the position and height of the maximum absorbance
and the associated stray light value Sλ.
In the second method "10 mm versus 10 mm H2O", the filter solution is measured
against water as reference. In both measurements, cells with a pathlength of 10 mm are
used. The absorbance measured at the certificate wavelength (at the edge of an
absorbance peak) must be greater than or equal to 2 A.
USP version 38, 40, 41
Test equipment / CRM Method Requirement
12 g/L KCl (190 to 205 mm) 10 mm versus 5 mm Sλ ≤ 0.01; Aλ ≥ 0.7 A

10 g/L NaI (210 to 259 nm)
Acetone (250 to 320 nm)
10 mm versus 10 mm Aλmax ≥ 2 A
50 g/L NaNO2 (300 to 385 nm)
From USP version 42
Test equipment / CRM Method Requirement
12 g/L KCl (190 to 210 mm) 10 mm versus 5 mm Sλ ≤ 0.01; Aλ ≥ 0.7 A

10 g/L NaI (210 to 270 nm)
Acetone (250 to 330 nm)
10 mm versus 10 mm Aλmax ≥ 2 A
50 g/L NaNO2 (300 to 400 nm)
Sλ = 0.25 × 10−2𝐴𝐴𝜆𝜆 Sλ – Stray light

Aλ – Absorbance value of maximum with longest wave
178
Due to the lower stray light quotient of the SPECORD PLUS devices, , the measuring
range for NaI, Acetone, and NanO2, is extended further beyond the range as given in
Table 3 of USP <857> by a few nanometers (towards longer wavelengths).
In accordance with the versions of USP<857>, the following acceptance criteria are
given for the two processes.
10 mm versus 5 mm variant
 The wavelength of the longest wave maximum (λ) is greater than the certificate
wavelength - measurement uncertainty
 Sλ ≤ 0.01. A λ ≥ 0.7 A
10 mm versus corresponding reference in 10 mm variant

Based on the long-wave determination of the wavelength (λ), at which the absorbance
exceeds the 2.0 A value, the following applies:
 λ is greater than the certificate wavelength - measurement uncertainty
 Below the certificate wavelength - measurement uncertainty, no absorbance values
< 2.0 A
Resolution USP requires that the spectral bandwidth of the spectrometer (SBW) used is not greater
than 1/8 of the natural half bandwidth (HWB) of the analyte absorbance.
For proof in the UV range, a 0.02% solution of toluene in hexane is measured against
hexane as a reference and the ratio of the absorbance maximum at 269 nm and the
absorbance minimum at 266 nm calculated. For most pharmaceutical applications, a
spectral resolution of 2 nm will suffice, which corresponds to an acceptance criterion for
the quotient A269/A266 of 1.3. It should be noted here that the quotient is temperature-
dependent and the influence of the temperature increases with the rising spectral
resolution.
Temperature dependency of quotients A269/A266 for toluene in hexane
Measurement Spectral bandwidth [nm]

temperature
0.5 ± 0.1 1.0 ± 0.1 1.5 ± 0.1 2.0 ± 0.2 3.0 ± 0.2
[°C]
20 ± 1 2.4.-2.5 2.0.-2.1 1.6.-1.7 1.3.-1.4 1.0.-1.1

25 ± 1 2.3.-2.4 1.9.-2.0 1.6.-1.7 1.3.-1.4 1.0.-1.1
30 ± 1 2.1.-2.2 1.8.-1.9 1.5.-1.6 1.3.-1.4 1.0.-1.1
Alternatively, suitable atomic emission lines can be measured. The spectral bandwidth
of the spectrometer corresponds to the half width determined for the atomic line. The
deuterium emission lines may be used for the Vis range. The mercury emission lines are
suitable for both the Vis and the UV range. (The line tables are saved in the validation
software of ASpect UV).
179
Test equipment / CRM Requirement
Hg and D2 emission lines The spectral bandwidth of the spectrometer (SBW) should be
≤ 1/8 of the substance absorbance band half width (2 nm will
suffice for most pharmaceutical substances).
Toluene in hexane A269/A266 ≥ 1.3
At the start of the validation, the parameters and test equipment need to be selected.
Execution
 In the main window, select Instrument  Test  Validation  USp.
 In the Settings window, select the ranges and parameters to be checked and make
the necessary settings for the test equipment entries (see below).
 Confirm the validation parameters with [OK].
 The Settings window is closed and the [Start] button released.
Tip
Once you have set all parameters and entered the required CRM data, save the data by
selecting File  Save or File  Save as in the Validation regarding USP window. The
data will then also be available for another validation, once the same CRM is used.
Selection of CRM and The USP requires the validation of the spectrometer in the operating range. Not only
measuring ranges must a suitable CRM be selected, often also the measuring range, e.g. the wavelengths
when testing the wavelength accuracy/precision, must be determined. If this is
necessary, the following button appears after selecting the CRM […].
Execution
 On the General page, make the necessary entries and mark the test parameters to
be validated with a tick.
 On the left side of the Settings window, select the test parameter.
 On the test parameter page, mark the CRM with a tick.
 Click on […].
 Mark the measured values in the selection window. If necessary, use [Settings] to
open the window Settings CRM and transfer the CRM data from the certificate to
the corresponding input fields.
Note: With CRM, on the pages Straylight and Resolution click immediately on
[Settings], there are no measured values to be selected here.
 Close the selection window with [x].
Tip
Several CRMs may be activated for a test parameter, thus covering your operating range.
180
Selection of a CRM and the measuring ranges
CRM data for a holmium oxide filter for validation of the wavelength
General page On the General page, determine the scope of the device validation and enter the
general parameters.
181
General page of Settings window
Title Title of the document

The title is displayed in the tab of the document (→ "The document window"
p. 15).
Operator Name of the user
If the usermanagement is installed, the user logged on into ASpect UV is
displayed.
USP version Selection of USP version for the validation
Part Selection of validation ranges
Slit For SPECORD 210 PLUS and SPECORD 250 PLUS

Spectral width (select spectral resolution of the spectrometer)
Note: Select the spectral width that you will also be using for your
measurements. The spectral resolution of the spectrometer should not
exceed 1/8 of the analysis band half width (→ "Test parameters, acceptance
criteria and required test equipment" p. 177).
Temperature Ambient temperature of installation location

Some CRMs are temperature-dependent.
Number of Required number of signature fields for test reports
signs Two signature fields are provided as standard (for the user and the service
technician). It is possible to select up to 4 signature fields.
182
Note Additional note for validation
Wavelength page On the Wavelength page, select the parameters for checking the wavelengths (accuracy
and precision).
Note: The filter wavelengths shown are only preset and give an approximate guide
value. Target values are the values from the certificates entered in the Settings CRM
window.
The following CRM may be selected:
CRM Wavelengths [nm]
Holmium oxide (glass 279; 360;453; 536; 637

filter)
Didymium oxide (glass 329; 472; 512; 681; 875
filter)
Holmium oxide solution 241; 249; 278; 287; 333; 345; 361; 385; 416; 451; 467; 536;
640
Didymium oxide 731; 740; 794; 801; 864
solution
Rare earth metal oxide 201; 211; 222; 239; 252
solution for lower UV
D2E lines 486; 656,1
Hg emission lines 194,2; 237,9; 248,2; 253,7*; 265,4; 289,4; 296,7; 302,15;
334,15; 365,0; 404,7; 407,8; 435,8; 546,1; 577,0; 579,1; 871,7;
1014,0; 1092,1
*Only possible with slit with smaller than 2 nm
Alternative CRM
Note: The wavelengths entered for the filter are default settings. Once they are
activated, the data must be taken from the standard certificate.
Absorbance page On the Absorbance page, select the parameters for determining the photometric
accuracy and precision. Here, the wavelength and absorbance range of your analyses
must be covered.
The following CRM may be selected:
CRM Measuring range / Concentration Wavelength range

[nm]
Potassium 20 to 200 mg/l in increments of 235 to 257
dichromate 20 mg/l
Niacin 6; 12; 18 and 24 mg/l 213 and 261
Neutral glass filter 0.04 to 3.0 Abs 440 to 635
Alternative CRM
183
Average absorbance of potassium dichromate at the wavelengths 235 nm and 237 nm:
Concentration [mg/l] Absorbance at 235 nm Absorbance 257 nm
20 0.25 0.29
40 0.50 0.57
60 0.75 0.86
80 1.00 1.16
100 1.25 1.45
120 1.50 1.75
140 1.75 2.05
160 2.00 2.35
180 2.25 2.65
200 2.50 2.95
Average absorbance of niacin at the wavelengths 231 nm and 261 nm
Concentration [mg/l] Avarage absorbance
6 0.25
12 0.50
18 0.75
24 1.00
Stray light page On the Straylight page, select the CRM and the method for stray light measurement.
CRM Measuring range [nm]
KCl (12 mg/l) 190 to 205
NaI (10 g/l) 210 to 259
Acetone 250 to 320
Na NO2 (50 g/l) 300 to 380
The wavelength ranges specified for the filter correspond to the information given in
table 3 of USP <857>. Due to the lower stray light quotient of the SPECORD PLUS
devices, the measuring range for NaI, Acetone and NaNO2 is extended by a few
nanometers towards longer wavelengths.
Resolution page On the Resolution page, select the method and the corresponding CRM for determining
the spectral resolution. In addition to the toluene/hexane method, the spectral
resolution may also be determined using the half width of the atomic emission lines.
CRM Measuring range
Toluene/hexane UV (ratio A269/A266)
D2E emission 486 nm; 656.1 nm,

lines
184
Hg emission 194.2 nm; 237.9 nm; 248.2 nm; 253.7 nm*; 265.4 nm; 289.4 nm;
lines 296.7 nm; 302.15 nm; 334.15 nm; 365.0 nm; 404.7 nm; 407.8 nm;
435.8 nm; 546.1 nm; 577.0 nm*; 579.1 nm*
*Only possible with slit with smaller than 2 nm
Archive automatically On the Archive automatically page, you can set the data to be automatically saved,
page printed or exported during validation.
TXT and PDF formats may be selected for the export. The USP_komplett.lst report
template must be selected.
measurements!
consecutively.

Window with instruction to insert a filter
 After each partial validation step, the test results and acceptance criteria in
accordance with USP is shown in a window.
The CRM measurement can be repeated and the test continued or interrupted.
Option Description
[Yes] Repeat CRM measurement

The measuring values already saved for the current test item are discarded. An
entry is made in the validation audit trail
[No] Accept measured values and continue with the validation
[Cancel Cancel validation

test] Measuring values already taken are temporarily saved and shown in the main
window.
185
Display of the validation results during the course of the test
Display validation results Once a measuring range for a test parameter/CRM has been fully processed and
processing of the next range or a new test parameter/CRM starts, the results are shown
on the corresponding tab below. It is thus possible to view measurement results from
validation measurements that have already been completed while the validation is still
running.
Validation regarding USP document window
No. Description
1 Selection of test parameter
186
2 For every CRM that has been activated for validation of the test parameter, a tab is
created.
3 Selection of CRM measuring range
4 Spectra display of measuring range selected
5 Results for the test parameters/CRM selected in the upper tab

The Allgemein sheet contains the settings for the General page of the parameter
window.

the far right tab.
Sign document Execution

 Select File  Sign document in the document window.
 Click on [Login].
 Select the signature.
 You have the option of entering a comment in the Comment field.
 Save document with File  Save or File  Save as.
 The document has now been signed. There may be no further changes made to
the data.

 In the Open window, select the print template USP_komplett.lst in the folder
\\Users\Public\Documents\Analytik Jena\ASpect UV\Reports\Validation\USP.
Attention With an operational qualification, the printout is included in the OQ report.
13.4 Validation Maintenance Economic
The economic maintenance validation is carried out as part of a service, once a service
agreement has been concluded. With the conclusion of a service agreement, you acquire
the validation with which the basic functions of SPECORD PLUS are tested.
187
Economic Maintenance The Economic maintenance module window appears after selection of Instrument 
module window Test  Validation  Maintenance economic.
Economic maintenance window
Enter the parameters of the standard under [Setup]. The validation results are
displayed on a separate tab for each test parameter. The Results spreadsheet outputs
the digital test results in each case and the Measured values sheet a graphical
representation of the measured spectra. In the list Part of validation, a parameter group
for the display can be selected in each case.
The audit trail may be shown in a separate window. The results of the validation may be
saved, printed and opened again.
Attention Take note of the following information in order to achieve optimum validation results:
 Start the validation after a device warm-up period of 2 hours. During this period both
lamps of the SPECORD PLUS must be switched on.
Test parameters The following parameters are validated in the Economic Maintenance:
Transmittance zero Measurement with covered measurement beam path and open
point reference beam path in the range of 200 to 1000 nm
The minimum and maximum of the zero point transmittance of the
device are calculated from the result.
Baseline deviation Recording of the baseline in the absorbance mode in the range of
200 – 1000 nm
188
Photometric accuracy Test with a certified standard filter

(absorbance at 465 Determination of the difference of the measured absorbance in
nm) comparison with the specified target values of the gray glass filter
F2, F3 and F4 with 465 nm
Wavelength accuracy Measurement of the wavelength positions of the peaks of a holmium
with holmium oxide glass filter with known target values for the wavelengths in the five
ranges 270 to 285 nm, 350 to 370 nm, 450 to 485 nm, 530 to 545
nm and 630 to 645 nm
Spectral resolution of Determination of spectral half width at D2E line at 656.1 nm
half width of D2E line The measurement is carried out using the UV lamp (D2E lamp).
Test equipment The following standards are required for the Validation Economic Maintenance
 Certified filter kit for the photometric measurement of the Vis spectral range and
certified holmium oxide glass filter for the wavelength measurement (HELLMA
standard filter set)
At the start of the validation, the certification data for the test equipment to be used
must be selected.
Execution
 In the main window, select Instrument  Test  Validation  Eco maintenance.
Tab Description
General User and a note to validation

If the user management is installed, the user logged on in the software is
diplayed
Display of parameters to be verified
Aceptance Display of limit values and tolerances for the parameters to be tested
criterias
All other tabs Resource parameters (designation, test wavelength, target values)
Validation window.
Enter user – General tab
189
Settings / General window; enter name of tester and comment
On the Allgemein tab, you can enter a user name and a comment for the validation. You
will also find an overview of the test parameters. The scope of testing is determined by
the manufacturer.
View limit values – The Limit values tab contains the limit values set in accordance with the manufacturer’s
Acceptance criteria tab specifications.
View limit values for the validation, Settings / Limit values window

Enter the identification numbers and standard values of the resources used on the
standards used
respective tabs.
Photometric accuracy at 465 nm tab
Enter the data for the gray filter set
Test wavelengths and nominal Enter wavelengths and target values for the test in
absorbance value accordance with the datasheet.
190
Wavelengths tab
Name ID Identification number of the holmium glass filter used
Test wavelengths Target values of the spectral peak in accordance with the datasheet
Place filters and cells Note for the Validation

All filters and cells with standard solutions used, including references, must always be
measurements!
is released. The validation runs assisted by the program

Window with instruction to insert a cell
 After each partial validation step the test results and defined tolerances are
displayed in a window. With its buttons the test results window offers the following
options for the subsequent validation method:
Option Description

Display validation results After completing the validation, the results are displayed on the tabs of the Validation
window.
191
Economic maintenance window with results
permissible tolerances. On the Measured values tab, the spectra or measured values are
shown.

the far right tab.

 The data is exported to the txt-format.
192

 In the Open window in the folder \\Users\Public\Documents\Analytik Jena\ASpect
UV\Reports\Validation\Maintenance economic, select the "Compact.lst" or
"Detailed.lst" print template.
193
User administration and electronic signatures
14 User administration and electronic signatures

The optionally available add-on module ASpect UV FDA 21 CFR Part 11 includes the
user administration and provides an option for the electronic signing of measuring
results.
14.1 User management
14.1.1 Preinstalled users
The user administration of ASpect UV permits the flexible issuance of user rights. Each
user can be granted or denied access to the individual functions within the entire
functional scope of ASpect UV.
Users installed by default Some users with various access rights have been created in the installation of ASpect UV
FDA21CRF Part 11 that can be used directly or as an example for your own rights
management. Logins and passwords have already been assigned for registration of
these users, they can however be edited or deactivated.
Tip
Deactivate these users to define templates for user profiles with different roles using the
Transfer user rights function. These profiles may then be transferred to users that
actually exist.
AJ Service The AJ Service user is assigned all rights (combination of Administrator and
Researcher). The role is used by Analytik Jena for testing purposes during software
qualification. It may be deactivated by the Administrator.
Administrator The Administrator has full access to the user management and can create, edit and
transfer user profiles. He does not have access to the measuring modules and cannot
create methods or start measurements.
Login Admin
Password admin
User Level 1 (Researcher) The User Level 1 (Researcher) has unrestricted access to the functions of ASpect UV. He
can view the user administration but cannot create a new user profile or modify existing
user profiles.
Login User1
Password user1
User Level 2 The User Level 2 (TechAssistant) has access to the following functions:
(TechAssistant)
 Open results
 Opening methods
 Open modules
194
 Start measurements
 Analyze measurements
He may not:
 Create methods
 Perform device corrections
 View the user administration
Login User2
Password user2
User Level 3 (Laboratory The User Level 3 (Laboratory Supervisor) has access to the following functions:
Supervisor)
 Open results
 Opening methods
 Open modules
 Create methods
 Analyze measurements
 Start measurements in the quick measurement, from which results cannot be saved
He may not:
 Start measurements in the ASpect UV modules
 Test and correct SPECORD PLUS
 View the user administration
Login User3
Password user3
User Level 4 (Service) The User Level 4 (Service) has been set up for Service. It has access to the following
functions:
 Perform simple measurements
 Correct SPECORD PLUS
 Test SPECORD PLUS
 Start and analyze energy measurement
 View list of users
He may not:
 Open the modules Photometry, Spectrum,Colormetry, Thermometry or Kinetics
 Access methods and results of these modules
Login User4
Password user4
195
14.1.2 Functions in the user administration
The user administration can only be organized by a user with the appropriate rights. The
Administrator user is a default setting here (→ "Preinstalled users" p. 194). The users
User Level 1 (Researcher) and User Level 4 (Service) can view the entries in the user
administration but cannot edit them.
Open user administration Execution
 In the main window, select User  User management from the menu.
 The user administration is opened. Dependent on access rights, users can be
created and edited or only the list of users can be viewed.
User administration
window
Window User administration with pre-installed users

1 Menu bar 3 Properties of the user profile selected in the
2 List of created user profiles list of users
The menu bar includes all functions for creating and editing user profiles.
Button Description
Add new user
Create a new user profile.
Edit user
Modify the properties of a user profile.
User rights
For a selected user, allow or deny access to each function in ASpect UV.
Transfer the user profiles.
Reports
Create reports via the registrations in Aspect UV and the user administration.
Close
Close the user administration.
The list on the left side of the window includes all user profiles created. On the right-
hand side are properties of the user profile selected (name, login, activation of profile,
password change and profile validity).
196
14.1.3 Create and edit user profiles
Create a new user Only users with full access rights to the user administration can create a new user. You
may, for example, use the default user Administrator (→ "Preinstalled users" p. 194).
Execution
 In the main window, select User  User management from the menu.
 Click on [New user].

 Perform the configuration in the User settings window.
User settings window
Option Description
Real name Name of the user
Login User login
E-mail address User email address
Enabled Enable if the user profile is to be used.
Blocked Enable if the user profile must not be used. With a locked user
profile the user can no longer log in to the program.
Change password Enable if the user is to create his own password after the first login
with this profile.
This must also be enabled if the user has been reactivated
following a lock/expiry of validity by Admin.
Account is valid until Enable and configure an expiry date if the user profile is to be only
valid for a specific time period.
Validity up to and including expiry day 23:59
Password/Confirm Password and password repeat for the user login to the program
password
Temporary password Activate and set number of days if the password should only be
valid for a specific period of time.
Validity up to and including expiry day 23:59
 Confirm user settings with [OK].

 The new user profile has been created.
Modify a user profile The properties of a user profile can be modified later. This function may e.g. be used to
lock a user profile or reset a password.
Execution
 Select users from the user list in the window User management.
 Click on [Edit user].

 Change the settings in the User management window.
 The user profile has been modified.
197
14.1.4 Assign user rights
Each user can be granted individual access to the functions of ASpect UV.
Execution
 Select users from the user list in the window User management.
 Select User rights ▸ Edit user rights from the menu.

 On the left-hand side of the window Edit user rights, select the respective category
and, on the right-hand side, activate all functions to which the user should have
access.
 Individual rights are assigned to the user profile.
User rights window with assigned rights in the General category
Tip
The options in the category General are related to the main window of ASpect UV. If you
wish to withdraw or grant the right Method/Open results, these options must be
accordingly (de)activated in the individual modules.
Transfer user rights The user rights of a user profile may be transferred to another profile. In this way,
sample profiles/roles may be created with different access authorizations and these
assigned to new user profiles. The default user profiles User 1 - User 4 may be used as a
basis for these user profiles.
Execution
 Mark the target profile in the User management window.
 Select User rights  Transfer user rights from the menu.

 In the Transfer Rights window, select the outbound/role profile for the user rights
and confirm with [OK].
 The user rights of the profile selected are transferred to the target profile.
198
Special signature features Permission to apply a signature to a document must be granted separately to each user.
A distinction is made here between signatures that must be applied in a defined
sequence and signatures with no sequence.
Procedure
 Select the user in the user list in the User management window.
 Select the User rights ▸ Edit user rights menu item.

 Activate all signatures for the selected user in the Signatures category.
 If you activate a signature that is not in the signature sequence, the Signing outside
of signature sequence option must be activated in the Own user account category.
Only then can the user use this signature. Otherwise, the signatures will not be
available for this user, despite activation in the Signatures category.
Note: The Signatures category is only visible if signatures have already been created in
the Options / Advanced / Signatures window.
14.1.5 Creating reports on the user management
In the user management, various reports can be created and printed out.
 Audit trail of the user management
 List of registrations in ASpect UV
 List of all users
 Rights of selected user
The right to view a report must be activated for a user in his/her user rights (→ “Assign
user rights” p. 198).
Audit trail The audit trail documents all changes to the user profiles made by a user with
administrator rights.
Execution
 In the User management window, select Reports  Audit Trail  View from the
menu.
 Using the filter settings, the entries may be sorted, filtered and printed according to
Admin, changed user profile and date. To do this, change the settings and click on
[Reload].
 To print/export the entire Audit Trail, select Reports  Audit Trail  Print from the
menu and the "UM_Audit Trail.lst” report in the path
\\Users\Public\Documents\Analytik Jena\ASpect UV\Reports\User Management.
List of registrations The registration list documents all successful and unsuccessful user logins in Aspect UV
with the log-on and log-off times.
Execution
 In the Benutzerverwaltung window, select Reports  List of logins  View.
199
 Using the filter settings, the entries may be sorted, filtered and printed according to
users, login times and registration status (rejected, successful or logged off). To do
this, change the settings and click on [Reload].
 In order to print/export the entire list, select Reports  List of logins  Print from
the menu and the "UM_LoginList.lst” report in the path
List of all users The list of all users includes the user profiles created with information on status, date
created and the date of the last login in ASpect UV. The user list may only be
printed/exported.
Execution
 In the Benutzerverwaltung window, select Reports  List of all users  Print from
the menu and the "UM_UserList.lst” report in the path
Rights of selected user With the rights report, you can print/export an overview of the rights for a selected user.
Execution
 In the User management window, mark a user profile.
 Select Reports  Rights of selected user  Print from the menu and the
"UM_UserRights.lst” report in the path \\Users\Public\Documents\Analytik
Jena\ASpect UV\Reports\User Management.
14.1.6 Log off user
With the function Log off a user can log off from the running program ASpect UV.
Afterwards, another user or the same user can log on again. This right should be
enabled for each user.
To log off, it is necessary to close all document windows of the modules beforehand.
Execution
 In the main window, select User  Log off from the menu.
 The main window of ASpect UV is locked.
 To log in again click on [Login].
 Enter the login and corresponding password.
 The ASpect UV program is released for further actions.
14.1.7 Lock ASpect UV
The Lock ASpect UV function locks the running ASpect UV program until the same or
another user logs on again. The right to unlock by another user must be activated
beforehand for the corresponding user in the user administration.
Running measurements are processed to the end in the locked state.
Execution
 In the main window, select User  Lock ASpect UV from the menu.
200
 The main window of ASpect UV is locked.

 To log in again click on [Login].
 Enter the login and corresponding password.
 The ASpect UV program is released for further actions.
14.1.8 Passwords in ASpect UV
Validity criteria for The validity criteria may be configured in the window once authorization has been
passwords granted in their user profile.
You can configure the criteria for the validity of passwords in the Options / Adwanced /
Password management window if the authorization is enabled in your user profile (→
"Automatically archiving documents" p. 30").
Login After three incorrect login attempts the respective user account is blocked. The block can
only be lifted by a user with administrator level access rights.
Changing the password A user may change the password if this function has been released in his user profile.
Execution
 In the main window, select User  Change password from the menu.
 In the Password and Confirm password fields, enter the new password.
 The new password is accepted.
14.2 Electronic signatures
What is a signature? Documents can be signed in the optional ASpect UV FDA 21 CFR Part 11 add-on
module.
Signing a file applies a digital signature (the login data of the signing user), and also
restricts editing of the file. These editing restrictions affect data management,
measurements and/or analyses on the worksheets, and can be defined for each
signature. If the functionality of a file is restricted by a signature, these restrictions
cannot be lifted by applying another signature. Optionally, a required sequence can be
defined for signatures that must be maintained for the signing process.
The signature of a file affects a logged in user. This user can only use the file functions
permitted by both the file signature and the user permissions.
Permission to apply a signature to a file must be granted to each user separately in the
respective user profile. A user can be granted permission to use multiple signatures.
Creating signatures in the Signatures are managed in the Options / Advanced / Signatures window (→ "Creating
program signatures" p. 158). Signatures can be created, edited and deleted here. The sequence to
apply signatures can also be defined here.
Assigning signature Permission to use one or more signatures is granted in the user profile (→ "Assign user
permissions rights" p. 198).
201
Signing a document The user must be logged into ASpect UV to apply signatures.
Procedure
 Select File  Sign document in the document window.
 Click on [Login] in the Signature append window and log in with your login data.
 The user's signatures are displayed.
 Select the signature in the Signatures list.
 A comment can be added in the Comment field, which is confirmed with [OK].
 If any further saving of the document is restricted by the signature, a message
appears that the file must now be saved for the last time. After confirming the
message with [Yes], save the file in the Save as window.
 The document has now been signed by the user. All further editing of the
document is restricted in accordance with the selected signature.
Signature append window for signing a file.
Displaying signatures Signatures are displayed in the bottom part of the document window next to the tabs
for data analysis.
 To display signatures in the document window, select the File  Display signatures
menu item.
 To hide the signatures, click on in the Signatures tab, or select the File  Hide
signatures menu item.
Displaying signatures in a document
202
AJ File Protection
15 AJ File Protection
The optional software AJ File Protection protects data against intentional and
unintentional data manipulation, e.g., deletion or modification of data. A filter driver
allows directory access by authorized applications, access by other applications is
blocked. The functionality of virus scanners and professional replication, synchronization
or data backup software is not impaired if Microsoft standards are complied with.
AJ File Protection must be installed and configured by the system administrator. The
installation requires administrator rights.
A detailed description of the installation and configuration of the software can be found
on the installation CD.
Attention In combination with the separate rights for automatically saving and exporting, the AJ
File Protection software guarantees complete data privacy for method creation, data
acquisition and evaluation, and archiving.
203
WebApp
16 WebApp
This program can monitor the instrument state through the network and query the
current results. To utilize this functionality the instrument computer and the end
instruments intended for its use must be connected to each other over a suitable
network (Ehternet, WiFi). The configurations described in this section must also be
made by the network administrator.
16.1 Sent data
The following data are sent to the webApp:
Systeminfo  Instrument type

 Instrument name
 Name of the accessories used
 Software name
 Logged-in users
System status  current measuring status of the SPECORD PLUS

 Configuration of the active module
Results table  the updated row of the results table with the configuration currently applicable for
the module
16.2 Configuring the network for using the web server
The system administrator must configure the remote monitoring.

 Select the menu item Configuration  Options and change to the tab Advaced /
WebServer.
 Perform the following configuration:
Option Description
Activate web Activates the web server with the following settings
server
Permit opening Permits or prevents the transfer of the results table

results data
Port The TCP network port on which the web server accepts connections. This
port might need to be approved in the firewall of the instrument computer
Number of Number of network connections the web server permits simultaneously. At
concurrent least one connection will be permanently assigned for each connected end
connections instrument.
204
WebApp
Instrument list See section "Configuring the instrument list" p. 205.

To use the central instrument list the corresponding INI file can be
selected here.
Enable SSL Enables SSL encrypted network connections. The port used for the SSL
encryption connection corresponds to the port following the configuration chosen
above (e.g. 8081 non-encrypted – 8082 encrypted. Connections on 8081
are then forwarded from http to https on part 8082). Both ports must
have been approved in the firewall.
SSL certificate To use SSL encryption an SSL server certificate must be selected in PEM
format. This certificate should be issued for the host name or the IP of the
instrument computer to prevent warnings about invalid certificates in the
end instruments.
Window Options/Web server to configure the web server
16.3 Configuring the instrument list
If the WebApp of the instrument computer is opened from an end instrument, initially
not instrument is shown in the instrument overview. In addition to the option to add
instruments manually to the network in the WebApp configuration, the list of the
instruments available in the network can also be configured centrally. To this end an INI
file must be created in the following format:
[WebSockets]
numberOfEntries=2
host0=192.168.0.25
port0=8081
ssl0=false
205
WebApp
host1=aj.aas.pc01
port1=8081
ssl1=false
For every instrument available in the network that is to be monitored on end

instruments an entry must be available in the INI file consisting of host, port and ssl. The
entry numberOfEntries must match the number of specified instruments. In addition,
the entries must be provided with ascending number suffixes.
The host can be specified both as IP or as FQDN or host name (it must be ensured that
the end instruments can resolve this host in the network into an IP).
The port and the SSL entry must match the configuration of the corresponding
instrument software (see network configuration).
It is recommended to store the INI file for the instrument list on a central network and, if
several instrument computers are used, refer to it in their network configuration.
16.4 Supported end instruments
The monitoring of instruments through the network works in all current browsers (IE10,
Chrome, Firefox, Safari, Android Browser). Both PCs/notebooks and touch-screen
instruments (smart phones, tablets) can be used.
The following operating system versions are recommended:
Instrument/operating Version
system
mobile Apple at least iOS6
instruments
Android At least version 4 but as a minimum a version on which the

Chrome browser can be installed. When using older versions there
may be display problems in the Android browser potentially
making the application unusable.
16.5 Opening the WebApp from the end instrument
Dependent on the configuration the WebApp can be opened through a browser using
the following address.
If the computer name is "aj.aas.pc01" and SSL encryption has been disabled in the
network configuration and port 8081 has been set:
http://aj.aas.pc01:8081/
If the same configuration has been provided with SSL encryption:
https://aj.aas.pc01:8082/
206
Overview of revisions
17 Overview of revisions
Version Changes
00.15 First issue
01.16 New: Moduls Thermometry, Colorimetry
02.16 Change in Colorimetry: correction of used standards
03.19 New: datda export in CSV format

Change: sample sequences
Change: lamp change is available in the measurement parameters
New: Validation of SPECORD PLUS acc. Manufacturer specification, Ph. Eur.,
USP and Maintenance Economic
New: Correcting infinite absorbance
New: AJ File Protection
New: Reports in the user management for audit trail, logins, user profiles and
user rights
04.20 Change: Instrument validation according Ph. Eur. issue 10
New: Instrument validation according USP issue 42 is available
A Advanced functions in the User management
(01/2021) Advanced options for signatures and signatures sequences
Automatic archiving
Instrument validation according USP issue USP-NF 2021 is available
207
Supplement
18 Supplement
18.1 Supplement A - Default methods
The Aspect UV installation includes prepared methods for various applications. After a
standard installation these "*.AUV" files are stored in the following folders:
C:\Program Files (x86)\Analytik Jena\ASpect UV\Methods
and C:\Users\Public\Documents\Analytik Jena\ASpect UV\Methods
Dependent on the selected language the methods are installed in German or English
during the installation.
Enzymology The enzymology methods are suitable for analyses using test kits by R-Biopharm. The
test kits can be ordered on the internet at http://www.r-biopharm.com/.
The following accessories are intended for the measurements:
 8-cell changer
 15-cell carousel
The methods have been installed in the following folders:
 German
\\Aspect UV\ Methoden \Enzymatik\8xWechsler und
\\Aspect UV\ Methoden \Enzymatik\15xWechsler
 English
\\Aspect UV\ Methods \Enzymatic\8xcell changer und
\\Aspect UV\ Methods \Enzymatic\15xcell changer
The following methods have been installed:
No. Method Ger Method Eng Part no.
1 Acetaldehyd Acetaldehyde Roche- Yellow Line

10668613035
2 Cholesterin Cholesterol Roche- Yellow Line
10139050035
3 Citronensäure Citric acid Enzytec Generic Line
E1214
4 D-/L-Milchsäure D-/L-Lactic acid Enzytec Generic Line
E1255
5 D-3- D-3-Hydroxybutyric Roche- Yellow Line
Hydroxybuttersäure acid 10907979035
6 D-Äpfelsäure L-Malic acid Roche- Yellow Line
11215558035
7 D-Glucose D-Glucose Enzytec Generic Line
E1210
8 D-Glucose/D-Fructose D-Glucose/D-Fructose Enzytec Generic Line
E1245
208
Supplement
9 D-Sorbit/Xylit D-Sorbitol/Xylitol Roche- Yellow Line

10670057035
10 Essigsäure Acetic acid Roche- Yellow Line
10148261035
11 Ethanol Ethanol Roche- Yellow Line
10176290035
12 Glycerin Glycerol Enzytec Generic Line
E1224
13 Harnstoff_Ammoniak Urea/Ammonia Roche- Yellow Line
10542946035
14 Lactose/D-Galactose Lactose/D-Galactose Enzytec Generic Line
E1213
15 L-Äpfelsäure L-Malic acid Roche- Yellow Line
10139068035
16 L-Ascorbinsäure L-Ascorbic acid Enzytec Generic Line
E1267
17 L-Glutaminsäure Glutamic acid Roche- Yellow Line
10139092035
18 L-Milchsäure L-Lactic acid Enzytec Generic Line
E1254
19 Saccharose Saccharose/ Enzytec Generic Line
/D-Glucose D-Glucose E1246
20 Saccharose/ Saccharose/ Enzytec Generic Line
D-Glucose/ D-Glucose/ E1247
D-Fructose D-Fructose
21 Stärke Starch Enzytec Generic Line
E1268
22 Sulfit Sulfite Roche- Yellow Line
10725854035
Methods for biochemical The methods for biochemical analysis are intended for the use of passive accessories
analysis (adjustable holders for small microcells).
 German
\\Aspect UV\ Methods \Bio
 English
\\Aspect UV\ Methods \Life Science
No. Method German Method English
1 Absorption 205 nm, Faktor 31 Absorption 205 nm, Factor 31
2 Absorption 215 nm, 225 nm Absorption 215 nm, 225 nm
209
Supplement
6 Absorption 260 nm Absorption 260 nm
7 Absorption 280 nm, Faktor 1,38 Absorption 280 nm, Factor 1.38
8 Absorption 280 nm Absorption 280 nm
9 DNS-Reinheit DNA-purity
10 Kalb und Bernlohr Kalb and Bernlohr
11 Kalckar and Shafran Kalckar and Shafran
12 Scopes-Formel Scopes-Formula
13 Wartburg-Christian-Formel (DNS) Wartburg-Christian-Formula (DNA)
14 Wartburg-Christian-Formel Wartburg-Christian-Formula (Protein)

(Eiweiß)
15 Whitaker und Granum Whitaker and Granum
Water analysis The methods for water analysis are suited for analyses with Spectroquant test kits. The
test kits can be ordered on the internet at http://www.merckmillipore.com/.
Passive accessories (standard cell holders) are intended for the measurements:
 German
\\Aspect UV\ Methods \Wasseranalytik
 English
\\Aspect UV\ Methods \Wateranalysis
No. Method Ger Method Eng Part no. Area
1 Ammonium Test Ammonia test 1.14752. 0.05-3.00mg/l

NH4-N
2 Nitrat Test Nitrate Test 1.09713. 1.0-25.0mg/l NO3-

N
3 Phosphat Test Phosphate Test 1.14848. 0.05-5.00mg/l
PO4-P
4 Sulfat Test Sulfate test 1.14791. 25.0-300mg/l
SO42-
5 Sulfit Test Sulfite test 1.01746. 1.0-60.0mg/l

SO32-
210
Supplement
18.2 Supplement B - Generation of measured values and recording of

reference values
This section provides a brief introduction to the calculation of measured values in the
spectral photometer and the meaning of the baseline and its correction.
For an absorbance measurement the transmitted energy I0 (reference value) and the
energy I weakened by the sample is measured at each wavelength of interest. This takes
place in arbitrary units, because only the ratio is of relevance. Quantitatively, the
absorbance behavior is expressed in transmittance T[%] or absorbance A:
I P Permeability
D=
I0 I0 Energy without weakening (base value)
I Energy exiting the sample
T[%] = D ∗ 100 T[%] Transmittance in percent
A = − lg D A Absorbance
Calculation of measured To reduce measurement errors through changes in energy during the measurement
values for a double-beam (drift), two beam instruments have been designed. With the aid of a reference beam the
spectrometer lamp spectrum is being "monitored". The two compared energy values from the
measurement and reference beam path compensate for the fluctuations of the lamp
energy.
Calculation of the reference value (also baseline):
IMR PR Permeability for the reference

DR =
IVR IRB Energy of the reference beam
IMB Energy of the measurement beam
Calculation of the sample value
IMP PP Permeability for the sample

DP =
IVP IRB Energy of the reference beam
IMB Energy of the measurement beam
Calculation of the measured value
DP P Permeability
D=
DR PS Permeability for the sample
PR Permeability for the reference
What to do with the In a single beam instrument the use of the reference sample is easy. Insert the reference
reference sample in sample into the beam path and perform the reference measurement. Replace the
double-beam reference sample with the sample containing the analyte and perform the sample
instruments? measurement.
In the double-beam instrument there exist, dependent on the subsequent sample
measurement, various combinations for the use of the reference and therefore,
unfortunately, also possible errors:
211
Supplement
The following 3 combinations are possible and lead to correct results:
Combination Reference measurement Sample measurement
Measuring beam Comparison beam Measuring beam Comparison beam

path path path path
1 Reference blank Sample blank
2 Reference Reference Sample Reference
3 blank blank Sample Reference
Correcting infinite Correction of the infinite absorbance is important for samples with very low
absorbance transmission and may contribute toward better measuring results for the absorbance.
With very low transmission values close to the 0% transmission line, the signal noises
cause a transmission of 0 or a negative transmission. Since the absorbance is calculated
as a negative decadic logarithm from the transmission (see above), and this is defined
only for values greater than zero, one usually sets the absorbance for these transmission
values to the magnitude of the calculation accuracy. In ASpect UV, this absorbance value
is 9.
Depending on the energy (spectral width and wavelength of the radiation source) and
the sensitivity curve of the detector (photodiode), the lower noise threshold in
absorbance is considerably lower. This results in high, unstable absorbance values, not
caused by the absorbance of the sample but by the signal noises and calculation
accuracy. Peaks or edges are poorly identified and evaluated here in these noisy spectra.
In order to correct this, the software is used to determine correction spectra, with which
the effect of the signal noise on the extent of the lower noise threshold is minimized. In
this way, it is possible to create absorbance spectra that are more easily analyzed, even
with very high absorbance values (5 to 7).
The correction of the infinite absorbance is required, in particular, for the stray light
tests according to USP. When evaluating the stray light using the "10 mm versus 5 mm"
method, a positive effect is achieved here, as there are two strongly absorbing samples,
as well as a reference and a sample.
18.3 Supplement C – Data format for importing sample information
Sample information data is imported in the method configuration dialog on the Sample
sequence tab.
Import file formats Data in the formats *.csv and *.txt. can be imported.
Keywords If the first row of the sample information files contains column information (separated
by a semicolon) all of the following rows will be assigned to the corresponding columns
by column. Keywords are not case sensitive.
The following keywords are valid for the columns:
Keyword Meaning
type sample type
212
Supplement
name sample name
nomvalue standard concentration
notes sample note
initweight sample weight
dilution dilution
varA, varB, … , var H Variable A, variable B, … , variable H
The following keywords are assigned to the sample types. Once again, the keywords are
not case sensitive:
Keyword Sample type
meas measurement
ref reference sample
wash flushing sample

calib calibration standard
nomeas no measurement
blank blank sample
Example
type;name;nomvalue;notes;initweight;dilution;varA;varB;varH;
ref;sample 1a;0;note44;55;66;1;2;
calib;standard1;55;note66;77;88;3;4;
calib;standard2;75;note66;77;88;3;4;
meas;sample2b;0;note55;66;77;2;3;
blank;blank value;0;note66;77;88;4;5;
wash;washing solution;0;note66;77;88;5;6;
Importing sample names If no keywords can be identified in the first row, all rows (incl. the first) are interpreted
only as sample names and assigned to the sample table in sequential order
Example
Sample 0815
Sample 0816
Sample 0817
Sample 0818
Sample 0819
213

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Operating manual

Service Analytik Jena GmbH

General information http://www.analytik-jena.com

Documentation number 10-2210-002-23

Implementation of the Analytik Jena GmbH

© Copyright 2021, Analytik Jena GmbH

1 About this document .................................................................................................. 7

2 Install ASpect UV ........................................................................................................ 8

3 Starting and exiting ASpect UV .................................................................................. 9

4 What you should know about ASpect in advance .................................................... 11

5 Quick measurement module and online measurements ......................................... 49

6 Photometry module ................................................................................................. 52

6.3.2 Analyzing photometry data............................................................................................. 62

7 Spectrum module ..................................................................................................... 71

8 Kinetics module ........................................................................................................ 91

9 Thermometry module ............................................................................................ 108

10 Colorimetry module................................................................................................ 126

10.1.6 Colorimetry – Settings / Note ....................................................................................... 130

11 General functions of the SPECORD PLUS – Menu Device ...................................... 139

12 Setting ASpect UV in the Options window ............................................................. 152

13 Validating the SPECORD PLUS (optional) .............................................................. 161

14 User administration and electronic signatures ...................................................... 194

15 AJ File Protection .................................................................................................... 203

16 WebApp .................................................................................................................. 204

17 Overview of revisions ............................................................................................. 207

18 Supplement ............................................................................................................ 208

19 Index .......................................................................Fehler! Textmarke nicht definiert.

1 About this document

Program versions This description is based on the following program versions:

Install ASpect UV  Switch on computer.

3 Starting and exiting ASpect UV

3.1 Starting ASpect UV

3.2 Starting ASpect UV with a module or quick start

3.3 Exiting ASpect UV

With the software-assisted shutdown routine, the monochromator of the spectrometer

4 What you should know about ASpect in advance

4.1 ASpect UV program structure

4.1.1 Modules in ASpect UV

 ASpect UV Maintenance economic

4.1.2 The main window in ASpect UV

ASpect UV main window

Menu Subitem Description

File  Open method Open a saved method for a subsequent measurement

Open results Open a saved result in its module environment

Close Exiting ASpect UV

Settings  Hide menu Hide menu bar

Hide/show Hide launch bar

Module  Quick Start quick measurements at a selected wavelength

Photometry Open a new document in the module Photometry /

Thermometry Open a new document in the module Thermometry /

Instrument Info After initialization, display information on the device, the

Test Test the functionality of the SPECORD PLUS

Change Change password of logged-on user

About Program version

Symbol Functional description

Open Photometry module / Add new document

Open Spectrum module / Add new document

Open Kinetics module / Add new document

Open Thermometry module / Add new document

Open Colorimetry module / Add new document

Load saved method for next measurements

Open saved measured values and analyses

Initialize SPECORD PLUS