UVWIN Instruction Manual

UV/Visible Spectrophotometer
Version 5.0

1
 Foreword
1. Introduction
UVWin5.0 software is the new release version of software for the PG range of UV/Visible
Spectrophotometers and replaces UVWin 4.0. Its main functions are to control the spectrophotometer, take
measurements, analyse and process the data which has been collected. Compared to its former version,
UVWin5.0 includes added functionality such as administrator management, quality control, safety features,
and 3D display. UVWin5.0 is also designed to be GLP compliant including data file encryption and
instrument validation.

2. A brief description of the manual

The manual contains 9 chapters, covering Installation, User Interface, Instrument Control,
Measurement and Results Processing. It gives a complete description of the UVWin5.0 Software
applications and data processing. Below are the contents of each chapter:

z Chapter One “Software installation and configuration”. This gives you an introduction to the
operating environment and installation of UVWin5.0. You will learn how to install UVWin5.0 onto
your PC correctly.

z Chapter Two “Software introduction”. This gives you an introduction to the User interface, Function
Menus, Configuration, Communication ports and other settings.

z Chapter Three “Instrument Control”. This tells you how to control a UV-Vis Spectrophotometer with
UVWin5.0. You will learn how to use the functions required to control a spectrophotometer and how
to set up basic methods.

z Chapter Four “Photometry”. This chapter describes how to use the Photometric Analysis functions
and how to set up a simple method for fixed wavelength analysis. You will learn how to use the
functions relevant to photometric measurement and how to set up the method parameters.

z Chapter Five “Spectrum Scan”. This chapter describes how to use the Spectrum Scanning functions
required setup a method for scanning samples for qualitative analysis. You will learn how to perform
a simple scan by setting up the scan method parameters.

z Chapter Six “Quantitative”. This chapter describes how to set up a method for quantitative analysis
and how to setup calibration curves with standards using different curve fits.

z Chapter Seven “Kinetics”. This chapter describes how to use the Kinetics functions and how to
perform a simple rate analysis by monitoring the absorbance change with time. You will learn about
the methods relating to the Kinetics parameter settings.
z Chapter Eight “DNA Protein Determination”. This section gives you an introduction of how to use the
DNA Protein Determination functions and how to set up a method for analysing DNA.

z Chapter Nine “Graphic Processing”. This chapter focuses on how to process the spectral data.

z Chapter Ten “Data Export”. This chapter gives you an introduction on how to export data results to

z other file formats.

z Chapter Eleven “Administration”. This section will show you how to use the Administration
functionality of UVWin5.0. You will learn how to create a new user account, a new group and how to
set up procedures that give different accessibility levels to different users within a group.

z Chapter Twelve “Conclusion”. This is a brief conclusion to the manual.

3. Concept of the manual

UVWin5.0 is a universal software package for all of the PGI UV-Vis Spectrophotometers. Some of
the instructions may differ from model to model. Therefore the manual explains all the functions of the
software which deal with all of our instruments. You should be aware of this and hence read the manual
carefully before using the instrument.
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Contents
Chapter 1 Software Installation and Configuration……………………………………………………1
1.1 Installation of UVWin5.0 Software……………………………………………...……………………1
1.2 UVWin5 Configuration ……………………………………………………...………………………2
1.3 Chapter Summary………………………………………………………………….…………………4
Chapter 2 Software Environment…………………………………………...………….…………………5
2.1 Startup of UVWin5.0………………………………………………………….…….…………………5
2.2 Getting Familiar with the UVWin5.0 Layout……… … …………… ……………6
2.3 Setting UVWin5.0 Operating Environment…………………………..…………….…………………10
2.3.1 Environment Options……………………………………………..………….…………………10
2.3.2 User Interface Layout……………………………………………….……….…………………12
2.3.3 Custom Tool………………………………………………………………….…………………12
2.3.4 Communication Port………………………………………………………….…………………13
2.4 Chapter Summary………………………………………….……………………….…………………14
Chapter 3 Instrument Control…………………………………………………….…………………15
3.1 Instrument Performance…………………………………………………………….…………………15
3.2 Energy Setting……………………………………………..……………………….…………………16
3.3 Baseline Correction………………………………………………………..……….…………………17
3.4 Dark Current Correction…………………………………………….…………….…………………17
3.5 Wavelength Correction……………………………………………….…………….…………………17
3.6 Goto Wavelength…………………………………………….…………………….…………………17
3.7 Setting Accessories…………………………………………………………….….…………………18
3.8 Instrument Initialisation…………………………………………………………….…………………19
3.9 Instrument Validation………………………………………………..…………….…………………19
3.10 Chapter Summary………………………………………………………………….…………………21
Chapter 4 Photometric Measurement…………………………………………….…………………22
4.1 A Brief Introduction to Photometric Measurement…………………….………….…………………22
4.2 Setting Photometric Measurement Parameters…………………………………….………………22
4.3 Photometric Measurement………………………………………………..……….…………………26
4.4 Saving and Printout ……………………………………………………….……….…………………26
4.5 Chapter Summary…………………………………….…………………………….…………………31
Chapter 5 Spectrum Scanning …………………………………………….…….…………………32
5.1 A Brief Introduction to Spectrum Scanning………………………………...…….…………………32
5.2 Setting up the Spectrum Scan Method Parameters……………………………….…………………32
5.3 Spectrum Scanning…………………………………………….………………….…………………34
5.4 Viewing Spectrum Information…………………………………………………….…………………34
5.5 Saving and Opening Spectrum Scan Files……………………………………...…………………34
5.6 Chapter Summary………………………………………………………………….…………………35
Chapter 6 Quantitative Analysis………………………………………………….…………………36
6.1 A Brief Introduction to Quantitative Analysis ……………………...…………….…………………36
6.2 Setting up the Quantitative Method Parameters………………………………….…………………36
6.3 Quantitation………………………………………….…………………………….…………………41
6.4 Saving Measurement Files………………………………………….…………….…………………43
6.5 Chapter Summary………………………………………………………………….…………………43
Chapter 7 Kinetics………………………………………….…………………….…………………44
7.1 A Brief Introduction to Kinetics……………………………………..…………….…………………44
7.2 Setting Kinetic Method Parameters……………………………………………….…………………44
7.3 Kinetics…………………………………………………………………………….…………………47
7.4 Saving Time Course Curves To File……………………………………………….…………………47
7.5 Chapter Summary…………………………….…………………………………….…………………47
Chapter 8 DNA Protein Determination…………………….…………………….…………………48
8.1 Use of the DNA protein determination function……………………..…………….…………………48
8.2 Starting the DNA protein determination function………………….……………….…………………49
8.3 Setting the measurement parameters….………………………………………….…………………49
8.4 Processing the Results…………………………………………………………….…………………50
8.5 Chapter Summary…………………………….…………………………………….…………………50
Chapter 9 Graph Tool Functions………………………………………………….…………………51
9.1 Use of the graph tool functions …………………………….………………….…………………51
9.2 Analysis of spectra………………………………………………………………….…………………51
9.2.1 Display settings……………………………………………………………….…………………51
9.2.2 General Observation……………………………………….………………….…………………53
9.2.3 Mathematical Calculations…………………………...……………………….…………………56
9.2.4 3D Spectra…………………………………………………………………….…………………58
9.3 Chapter Summary…………………………………………….…………………….…………………61
Chapter 10 Exporting Data…………………………………..…………………….…………………62
10.1 A Brief introduction to Exporting Data ………….……………………………….…………………62
10.2 Exportable Data……………………………………………………………….…………………62
10.3 Exporting Data ………………………………………...………………………….…………………63
10.4 Handling exported data files…….…………………………………….………….…………………64
10.5 Chapter Summary… ……………………………….…………………………….…………………64
Chapter 11 Administration Facility………………………………………………….…………………65
11.1 A Brief introduction to the Administration Facility…………………………………….……………65
11.2 Managing Users…………………………………………………………….…………………65
11.2.1 Setting up Users……………………………..………………….…………….…………………66
11.2.2 Group Management……………………………….………………………….…………………68
11.3 Security………………………………………………….………………………….…………………69
11.4 Operation Log ……………………………………..……………………………….…………………70
11.5 Other Administrative Functions…………………………………………………….…………………71
11.5.1 User-Defined Log………………………………..…..…….…….…….…….…………………71
11.5.2 Locking the System…………………………….…………………………….…………………71
11.6 Chapter Summary……………………………………….………………………….…………………72
Chapter 12 Conclusion……………………………………………………………….… ………………73
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Chapter 1 Software Installation and Configuration

Key points
In this chapter you will learn how to install UVWin5 onto your PC and how to use the Configuration
program to setup the software’s external parameters.
This includes:
z Installation of UVWin5.0 software
z Configuration program
z Chapter Summary

1.1 Installation of UVWin5.0 software

UVWin5.0 can run on Microsoft Windows 98/2000/XP/Windows 7.0. The minimum PC requirements
are 800MHZ CPU, 64Mb RAM, and CD-ROM or DVD-ROM drive.
The recommended configuration is a Pentium 1.4Gb CPU, 256Mb RAM, 32Mb Graphical Display
Card, 17 inch Color Display, CD-ROM or DVD-ROM. The Graph Display Card is mainly for the 3D
Display function within the UVWin Software. If you do not have a need for 3D Display then you can just
use the standard card that comes with the PC. UVWin5.0 requires at least 20Mb hard disk space so you
should ensure that you have enough hard disk space for the installation of the software.
UVWin5.0 installation is a simple step by step process. You should run the Setup.exe program from
the Installation Disk this will allow you entry into the Installation Program. Please refer to Figure 1-1.
Follow the prompts in the Installation Program to finish the setup. The serial number of the software is
recorded in the Installation Pack. Please input the serial number when you are prompted.

Figure 1-1 UVWin5.0 Setup Window

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After installing the software a shortcut is created on the desktop. Also a folder for the software is
created in Program Group on the Start Menu. The folder includes shortcuts for UVWin5.0, Configuration
program, and uninstall. Please refer to Figure 1-2.

Figure 1-2 UVWin5.0 Program Group and Shortcut

As UVWin5.0 is a universal software package for UV-Vis spectrophotometers, its installation

program may vary with different instrument models. As a result the shortcut and folder created by the
installation program may also vary.
If you need to remove UVWin5.0 from your PC, you can run the Uninstall program from the UVWin
5.0 Program Group. The Windows System will remove UVWin5.0 from your PC automatically. Please
note that before you remove the software, you should backup your measurement data in advance so as not
to lose it. The default folder for measurement data files is:
C:\Program Files\company\UVWin5.0\Data
This may vary with different instrument models. So you may need to find the relevant folder from the
UVWin5.0 shortcut on the desktop.

Tips：Shortcut

If you do not know the path for the file name that the shortcut links to, you can point at the shortcut
and click the right mouse button. From the pop-up menu, select Attribute to open the Shortcut Attribute
Window. In the Attribute Window, click the Search Object to open the Windows Explorer and go to the file
folder that the shortcut links to.

1.2 UVWin 5.0 Configuration

The UVWin5.0 Configuration Program is a tool for setting UVWin5.0 operating conditions. You can
start it by Selecting [Start]-> [Program]-> [UVWin5.0 Folder] ->[UVWin 5.0 Configuration]. The
operation menu for the Configuration Program is as shown in Figure 1-3.
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Figure 1-3 Setting the Communication Port

The Communication Port

To set the communication port between PC and instrument, choose a port and click the “Test” button
to test if the port exists and to show the test result. When it shows on this indicates that the port exists. If
this is not the case then please test another port. Please note that if the status shows that the port is online
this may mean that the port exists, but another program may be using it. So you will have to run UVWin to
confirm this.

The Accessories Tab

This is where you select the type of accessory you have installed in your instrument. The options are
“8-Cell Holder”, “5-Cell Holder”, “Fixed Cell Holder”, “Fixed Flow Cell Holder” and “Integrating Sphere”
as shown in Figure 1-4. Select the one which applies to your instrument.

Figure 1-4 Selecting the Accessory

Fixed Bandwidth Options

This is where you set the bandwidth of your instrument if it has a fixed slit, as shown in Figure 1-5.
The value will be kept with your measurement data files and it will also show on any printout.
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Figure 1-5 Selecting the fixed bandwidth

Help Tab

The Help Tab will guide you through the UVWin5.0 configuration program.

After you have finished the configuration, click on OK to save the settings and exit. If you want to
cancel an option then click Cancel or close the dialog box directly. If you select the option to Run UVWin
after exit, the system will run UVWin5.0 automatically after you exit by selecting OK.

1.3 Chapter Summary

This chapter gives an introduction to the UVWin5.0 installation procedure and the configuration
programme.
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Chapter 2 Software Environment

Key points
In this chapter you will learn about the UVWin5.0 user interface, the menu functions and setting the
environmental parameters.
z Startup of UVWin5.0
z Getting familiar with the UVWin5.0 Layout
z Setting the UVWin5.0 Operating Environment
z Chapter Summary

2.1 Startup of UVWin5.0

Before running UVWin5.0, make sure that the UV-Vis Spectrophotometer and the PC are connected
and the instrument is turned on. Then you can click on the UVWin5.0 shortcut, the startup window is
shown in Figure 2-1.

Figure 2-1 UVWin5.0 Startup Window

If the software has been correctly configured you should see the instrument initialisation window as
shown in Figure 2-2. (Please refer to 3.1 Instrument Performance for details on initialisation)

Figure 2-2 UVWin5.0 Initialisation

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Once initialisation is completed, (this takes approximately 3 minutes), the UVWin5.0 main user
window will appear. If initialisation fails, the system will display a message, as shown in Figure 2-3. Select
“Yes” to enter into the Software, or “No” to exit. Please note that should the initialisation fail then it is
advisable not to enter the Software to operate an instrument. Initialisation failures may be due to a variety
of causes and it may be possible that there are problems with instrument components. If you continue to
perform any analysis using potentially defective components, you may be at risk of further damaging the
instrument. So it is advisable to exit the Software should the initialisation fail. Then refer to the
“Instrument service manual” or contact a service engineer.

Figure 2-3 Initialisation Failure Message

If the instrument and the PC are not communicating then there is probably a problem with the
connection between the instrument and the PC. A message will be displayed as shown in Figure 2-4. You
should check if the communication cable is connected and that the instrument is turned on. Also the
communication port that you selected may be incorrect or incompatible. If this is the case you should
select Cancel to exit the program and run the UVWin5.0 Configuration programme to reset the
communication port and startup UVWin5.0 again (Please refer to 1.2 UVWin5.0 Configuration on how
to use this application).
If you select “Ignore” you can enter the Software offline but you will be unable to operate the instrument
in this case.

Figure 2-4 No communication between PC and Instrument

2.2 Getting familiar with the UVWin5.0 Layout

Once you enter the UVWin5.0 Software you will be presented with the main user interface. As Shown
in Figure 2-5.
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Tool bar Menu Toolbar

Tree Window

Status Bar Module Screen

Figure 2-5 UVWin5.0 Main User Interface

The Main User Interface adopts a multi-file tree structure layout. The software has a user friendly
operating environment and is designed to make it simple for all users to navigate. From the above figure
2-5, you can see that the UVWin5.0 main user interface is composed of the following parts: Menu
Toolbar, Toolbar, Status Bar, Tree Structure Window, and Module Window. Let’s have a look at these in
more detail.

Menu Toolbar

Most software packages have a Menu and from the Menu you can access a number of drop down
menus. The majority of functions in the software are provided in the Menu bar. The Menu bar includes
the following menus File, Edit, Measure, Graph, Mathematical Calculation, Administrate, Tools,
Application, Window, and Help. As shown in Figure 2-6. You can use menu functions provided in the
Menu bar to accomplish the corresponding actions as described by the software. In the following
chapters you will be given a detailed introduction to the menu functions.

Figure 2-6 UVWin5.0 Menu bar

Tip : Shortcut Keys

In Windows Systems there are many ways you can open a menu. Besides clicking a mouse, you can also
press the Alt Key and the underlined letters of the Menu words. For example, for File (A) Menu you can
press Alt+A Key to open File Menu. In addition, in some menus, there are specific tags after the headings,
such as Print(P) Ctrl+P, or Parameters Setting .. F4. Among others Ctrl+P and F4 are hot keys for
corresponding menus. What you can do is simply press the hot key to select the function of the
corresponding menu. So for Print(P) …..Ctrl+P Menu, just hold Ctrl key and hit P.
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Toolbar
The role of the Toolbar is to list those commonly used functions together as tool buttons. In this way,
users can the access the menu functions by a single press of the tool buttons, thus making the software
faster and easier to use. UVWin5.0 Toolbar consists of a General Toolbar, a Configuration Toolbar, a
Window Toolbar, a Graph Toolbar, a Control Toolbar and an External SW Toolbar, as shown in Figure 2-7.
Between each Toolbar, there is a partition line. When you move your mouse cursor over the partition line,
the cursor changes to the “ ” shape. At this point you can hold by pressing the left button of the mouse
and by dragging the partition line you can change its position. As shown in Figure 2-8.

General Toolbar Configuration Toolbar Window Toolbar

Control Toolbar Graph Toolbar Kit Toolbar

Figure 2-7 UVWin5.0 Toolbar

Toolbar Partition line

Figure 2-8 Dragging Toolbar

Status Bar

The Status Bar is located at the bottom of the window and it is used for displaying the current live
status of the software. It indicates the type of command which has been executed, data file type, the
current user name and the online / offline status. When using the Spectrum Scan or Kinetics application
the Status Bar also displays the actual coordinate values of the cursor location. As shown in Figure 2-9.
In addition, if your main interface window is not set to the maximum size, there is a “ ” sign at the
lower right of the Status Bar, you can drag this symbol with your mouse cursor to adjust the window
size.

Live coordinates Data file type Current user Line status

Figure 2-9 UVWin5.0 Status bar

Tree Type Window

The Tree Type Window structure is commonly applied in software design. Its clear framework is more
convenient to work with and its characteristics are more visual. In UVWin5.0 there are two tree type
windows. One is used for accessing the Applications windows and module files and is called the “Work
Space Window”. Another one is used for the display and control of instrument accessories called the
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“Accessories Window”, (Please refer to 3.7 Setting Accessories, as shown in Figure 2-10. A detailed
introduction to the “Accessories Window” will be introduced in the next Chapter.

Workspace Window

Accessories Window

Figure 2-10 Work Space Window and Accessories Window

Application Windows

The Application windows are sub-windows of the main interface window. Each application can
accomplish different measurement functions. The Application windows include the Photometry Window,
the Spectrum Window, the Quantitative Window, and the Kinetics Window. As shown in Figure 2-11. You
can enable each Application window by using the mouse to select the window Title Bar, or by selecting the
corresponding Application in the “Work Space” area.

Figure 2-11 Application Window

Tips : Window
UVWin5.0 adopts a multi-file structure. Each application module is capable of being resized and
moved or opened and closed, which makes its operation more flexible and convenient.
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2.3 Setting UVWin5.0 Operating Environment

The UVWin5.0 working environment enables the user to customise the working environment. This
includes setting the data format, file options, how the software responds to different commands,
configuring the Toolbars, Windows and Communication Port Settings. This makes routine operations
easier to perform and makes the software a convenient tool for setting up individual user preferences.

2.3.1 Environment Options

The Environment Options enable you to setup your preferred working environment. This includes
User prompt settings, File Options, Recent File List, Data format and how to set a Default Directory as
shown in Figure 2-12.

Figure 2-12 Environment Options

Indication (Prompts)

Prompt before Measurement: When you press the Measurement Button, a prompt message will
display, informing you to put in the sample. After you have inserted your sample then press OK and the
measurement is taken.
Prompt after Measurement: This is similar to the prompt before measurement, this time a prompt
message will be displayed after the measurement to indicate that the measurement has been completed.
Beep after Measurement: Similar to Prompt after Measurement, but instead of displaying a prompt
window, a beep will be made after each measurement to remind you that the measurement has been taken.
If your PC is fitted with an audio card, speakers or an earphone, the sound will be sent out from the
speakers or earphone. If there is no audio card in the PC, the PC speaker will send out a short beep.

File Option

Maximum open files: You can set the number of files that each application module can hold
simultaneously. For example in Spectrum Scan it is the maximum number of spectra which can be
displayed at the same time.
Auto-Save Baseline as a file: This function is designed only for the Spectrum Scan Module. If you set
this function to on the system will save the baseline data to a disk file automatically after each baseline
correction finishes. The software will load the saved baseline data automatically when UVWin 5.0 runs
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next time. This way you can perform a spectrum scan directly without doing baseline correction first.
In general a full range spectrum scan can be relatively time consuming and a baseline correction can
take nearly the same time as that of a spectrum scan. So by saving the baseline data you not only save time
on your workload but also reduce the wear and tear of the instrument. On the other hand it is possible that
external working conditions and environmental factors may affect the status of the UV-Vis
spectrophotometer. If the scan result is not satisfactory then you will have to perform a baseline correction
again.
However for UV/Vis Spectrophotometers using CCD detectors where fast scanning occurs then the
above analogy no longer applies. A UV/Vis Spectrophotometer’s CCD detector receives spectra reflected
directly by the grating and CCD detectors are very sensitive to small changes in environmental conditions.
Therefore it is important that an instrument baseline correction is performed before the collection of any
data.

Recent File List

The Recent File List keeps a record of the most recent data files for future reference. The recorded
filenames are listed in the sub menu of the File Menu. If you want to view one of them just click on the
corresponding menu item.
Data File: All measured data files.
Method File: All measured parameters files.
Work Space: All Work Space Files.
If you want to clear recent file records, you can press Clear All to delete all records.

Data Format

Data Format allows you to set how the data is displayed in UVWin5.0. This is to show how many
decimal places to use. For example “0.00” is two decimal places or digits for display. The data you can set
its decimal places include:
Abs: Absorption
T%: Transmittance
E%: Energy
Conc: Concentration in Quantitative measurement.
SD: Standard Deviation in Quantitative measurement.
RSD: Relative Standard Deviation in Quantitative measurement.
R2: Standard Linear Relativity.

Default Directory

You can set a default directory location on your PC for accessing your data and method files. This will
be the original directory displayed in the dialog box when you save or open a file. “Data” indicates the
directory for all of the data files you save. “Export” represents the default directory for exporting files. You
can select different directories by pressing “…” buttons.
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2.3.2 User Interface layout

The UWWin5.0 user interface layout can be customised to your own preferences, this mainly includes:
Toolbar, Status Bar, Work Space, and the Accessories Window. You can select the Custom Layout from the
Window Menu as shown in Figure 2-13.

Figure 2-13 Custom Layout

In the Custom Layout window you can decide to display or hide up to 6 toolbars and choose to hide
or display the Work Space window, the Accessories window and the Status Bar (please refer to 2.2 Getting
familiar with UVWin5.0 Layout for more details about Toolbar, the Work Space, the Accessories window,
and the Status bar).
There is a simpler way to select the toolbar options you can do this by moving your mouse cursor
over the main toolbar and then click the right button of the mouse. A menu will pop up as shown in shown
in Figure 2-14. You can click items with “a” mark to show corresponding toolbars. Alternatively you
can select the Window menu and go to Toolbars to view the same options.

Figure 2-14 Toolbar Menu

2.3.3 Custom Tool

You may have noticed that one of the Toolbar options is named Tool. This toolbar has the function to
open external application programs, such as MsExcel, MsPaint and Calculator etc. You can select the
option called Configuration Tool from the Tool Menu to setup the “Tool” functions. The Configuration
Tool is shown as Figure 2-15.
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Figure 2-15 Configuration Tool

The left window shows the list of programs or tools currently available while the right side shows the
relevant tool information. Tool information contains the file path name and location of the program on your
computer.
z Program: This is the path for the executable file for the highlighted program. This could be an
executable file (.EXE) or a page file (HTM). You can click on the button to the right of the
program select the program file.
z Title: This is the title of the program highlighted.
z Directory: This is the working folder for the program. The default working folder is the
directory in which the program is saved. When you select a program file the system will set this
to be the default.
z Parameters: Operation parameters of the highlighted program. For example if you want to setup
a tool to open the “hh.txt” file in C Disk root directory, with Windows Notepad. You should set
the Program as “C:\Windows\Notepad.exe”, and Parameters as “C:\hh.txt”.
z Icon file: This is the Icon that is used to display the program in the Toolbar. The default icon is
the program file. You also are able to click on the button to the right of the icon file and
select a different icon.
z Icon: For an icon file of “ICO” extension name, it only includes one icon. But for a file of “EXE”
or “DLL”, it may include many icons. So you can select other icons in the icon file using this
option.
If you want to add a new program tool, you can do it by editing the tool information directly by
selecting the Add button. After you finish adding a new program you can press “Replace” to update it. The
“Delete” button is used for deleting a current tool. After you finish the tool configuration you can close the
window and the icons in the “Tool” toolbar of the main interface will be refreshed.

2.3.4 Communication Port

The Communication port is the only interface for the PC to instrument connection which is essential
for instrument control. Therefore, if the communication port is set incorrectly, it cannot accomplish
communication with the instrument. If you need to change the communication port settings, you can select
the Communication Port submenu under Tool Menu. The Communication Port Setting Window will pop
up, as shown in Figure 2-16.
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Figure 2-16 Communication Port Setting Window - Online (Left); Offline (Right)

You can select a port name in the pull-down menu of the Port option, and then press the OK button. If
you are not sure which port connects with the instrument, you can click on the Auto-Test button and the
system will test all ports automatically. If it finds the port which is connected to the instrument it will go
online automatically.

Tips: “Serial Interface”

There are several versions of Serial Communication Interfaces these depend on the type of usage and
development. They all come from the improvement of the Standard RS-232. The RS-323C Standard is a
Communication Protocol released in 1969, which was developed by the American EIA (Electronic
Industries Association), BELL, and some other companies. It is designed to communicate data at a
transmission rate of 0-20000b/s. The standard defines relevant aspects of serial communication interface,
such as the function of signal lines and electronic characteristics. The majority of equipment manufacturers
produce their communication devices compatible with the RS-232C, so it has been adopted widely as a PC
Communication Interface.

2.4 Chapter Summary

This chapter is an introduction of the UVWin5.0 working interface and environmental settings. You
can setup the UVWin 5.0 operating environment by using the Environment Setting functions. In addition,
you can also make future operation easier by utilising the Custom Tool to setup the Toolbars. After reading
this chapter, you now have the fundamental knowledge on how to start using UVWin5.0. In the next
chapter you will learn about the other functions of UVWin5.0.
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Chapter 3 Instrument Control

Key points
In this chapter we will cover the following: Instrument parameter settings, instrument correction,
instrument validation and instrument accessories.
It includes:
z Instrument performance
z Energy setting
z Baseline correction
z Dark current correction
z Wavelength correction
z Goto wavelength
z Accessories setting
z Chapter Summary

3.1 Instrument Performance

For UV-Vis spectrophotometers the ability to select the correct optical parameters is of paramount
importance. Today’s instruments are capable of being fully automated there are many additional optical
settings that make operation easier and control more flexible. The UVWin5.0 Instrument Performance
Option provides you the ability to select instrument optical system parameters. You can select the
“Instrument Performance” menu from the submenu under the “Measure” menu or by clicking on the
toolbar button . The software will display the “Instrument Performance” setup window. Here you can set
the status of the light source and change some of the other characteristics of the optical system. As shown
in figure 3-1.

Figure 3-1 Instrument Performance Window

z Lamp Status
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This allows you to switch the Deuterium and Tungsten lamp on and off. It is on when the button is red
and it is off when the button is grey.
z Spectral bandwidth
Here you can choose the spectral bandwidth or the instrument slit width. The spectral bandwidth
options are relative to the type of instrument you are using. For example a UV-Vis split beam
spectrophotometer has a spectral bandwidth optional range of 0.5nm, 1.0nm, 2.0nm, and 5.0nm. But a
UV-Vis double beam spectrophotometer with a PMT detector might have a spectral bandwidth optional
range of 0.1 nm, 0.2 nm, 0.5nm, 1.0nm, 2.0nm, and 5.0nm. For fixed bandwidth instruments there is no
need to set this parameter. If you want to change the spectral bandwidth of a fixed bandwidth instrument
then please refer to 1.2 Configuration Program.
z Response time
Here you can set the instrument sampling response time. The longer the response time then the more
accurate data can be collected for the sample. This parameter is only applicable to general UV-Vis
spectrophotometers and not to CCD UV Spectrophotometers.
z Lamp interchange wavelength
Here you can set the wavelength interchange for the deuterium and tungsten lamp. If you select a
wavelength which is less than the interchange wavelength then the instrument will changeover to the
deuterium lamp automatically. If you select a higher wavelength than the interchange wavelength it is
considered to belong to the visible range and the instrument will change over to the tungsten lamp
automatically. This parameter only applies to general UV-Vis spectrophotometers. For CCD Fast UV
Spectrophotometers there is no such parameter.

3.2 Energy Setting

Generally for UV-Vis spectrophotometers the photometric mode includes absorption (Abs) and
transmittance (%T) measurements. The Energy mode is sometimes required to analyse the energy
distribution of samples. In this case the photometric mode is required to be set to energy mode. For energy
mode there are some special settings. After you choose Energy Mode you can select the Energy submenu
under Measure Menu or click on tool button, to open the energy settings window. As shown in Figure
3-2.

Figure 3-2 Energy Setting Window

z Lamp
Here you can select the lamp to be used in energy measurement mode. For general UV-Vis
spectrophotometers it is set to deuterium lamp or tungsten lamp. However for an instrument using a
see-through type deuterium lamp there is no need to set the lamp for energy mode.
 17

Tips: “see-through type deuterium lamp”

The See-through type deuterium lamp is an innovative technology for light sources which is on the
increase. Its principle is to utilize an open aperture in the center of the deuterium lamp and to allow the
light from a secondary source to pass through the same light path as the deuterium lamp. With this
technology you can leave out an optical mirror using the old light source and adopt a straight-line
arrangement of the tungsten lamp and deuterium lamp.

z Gain
Here you can set the gain value under energy mode. The gain value range varies with different
instrument models. For example a UV-Vis split beam spectrophotometer has a gain value range of 0-15
with 16 steps. A UV-Vis double beam spectrophotometer with a PMT detector has a gain value range of
1-7 with 7 steps. The role of the gain is to amplify the measured signal to improve the measurement on
spectra of low energy. However gain amplification can affect the data quality since a higher gain setting
can also amplify any noise that the data signal has. After you have altered the energy setting you can press
the OK button and the system will update the new energy parameters.

3.3 Baseline Correction

A Baseline correction can be performed in the absorption mode or in transmittance mode. Also it is a
typical correction function of spectrum scanning. Measurement under the two photometric modes requires
correction using a blank solution or blank solvent, so baseline correction should be performed before
scanning. Also a baseline correction should be performed after changing the scan method or parameter
settings.

3.4 Dark Current Correction

The Dark current correction corrects the instrument circuit noise. You are required to put a black
block into the cell holder to perform a dark current correction.

3.5 Wavelength Correction

The Wavelength correction allows you to correct the instrument wavelength accuracy. Any large
deviations in wavelength can have a direct influence on the measured results. Therefore it is necessary to
use wavelength correction to adjust it.

3.6 Goto Wavelength

Goto wavelength allows you to set the current instrument wavelength. Select the Goto Wavelength
submenu under the Measure Menu to open the Goto Wavelength Window as shown in Figure 3-3. In this
window input the wavelength you want to set and press the OK button to confirm it.
 18

Figure 3-3 Goto Wavelength

3.7 Setting up Accessories

The Accessories setting menu allows you to setup any accessories that the instrument is using. At
present the optional accessories include the Fixed Cell Holder, Flow Cell, Integrating Sphere, 5 Cell
Holder and 8 Cell Holder. Select the Accessories submenu under Measure Menu to open the Accessories
window as shown in Figure 3-4. In this window you can select the type of accessories and setup the sample
names accordingly. If you select the multiple cell holder you can click on a Position “ ” to setup the
current sample cell.

Figure 3-4 Accessories Setting Window

If one cell is set as a Blank the system will move to it and perform a zero when the autozero or
baseline correction is selected. If a cell position is set to “Not in use” then the system will not move to it
during measurement.

Sample Name

By double clicking on the sample you can type the sample name of each corresponding cell. The
name will be saved with the measurement file and can also be printed out.
 19

3.8 Instrument Initialisation

You can select the Initialisation submenu from the Measure Menu to perform an instrument
initialisation again, as shown in Figure 3-5. From the initialisation window UVWin5.0 Software can
initialise some of the instruments automated mechanisms. The automation mechanisms may vary with
different instrument models. For example instruments with variable slits will initialise the slit motor, whilst
instruments with a fixed slit don’t. The UVWin5.0 initialisation window covers the whole screen and
displays information about the software. The initialistion window shows the instrument name, instrument
serial number, and the tests that will be performed. During the initialisation procedure, passed items show
“ ” sign, whilst failed items show “ ” sign. Please refer to the instrument service manual for detailed
information on troubleshooting initialisation failures.

Figure 3-5 Instrument Initialisation

3.9 Instrument Validation

The Instrument validation programme will help you accomplish instrument performance checks
automatically. To do this select the Instrument Validation submenu from under the Measure Menu as
shown in Figure 3-6.

Figure 3-6 Instrument Validation Window

 20

Instrument Validation includes six tests: “Wavelength accuracy”, “Photometric accuracy”, “Stray
light”, “Noise”, “Baseline flatness”, and “Stability”. You are able to select each of them and set them up.

Wavelength Accuracy

This test uses the characteristic spectral lines of a “Holmium Oxide filter” or the “Deuterium lamp
emission lines” to check the instrument wavelengths. You can click on the Setting button to make small
amendments to the wavelengths to be used.

Photometric Accuracy

This test uses the characteristic spectral lines of a “Neutral density filter” or “Potassium dichromate
solution” to calibrate the instruments photometric accuracy. You also can click on the Setting button to
make small amendments to the characteristic absorption.

Stray light

This test uses “Sodium Nitrite solution at 340nm” or “Sodium Iodide solution at 220nm” to check the
instruments stray light.

Noise Level

This test measures the instruments noise level in 120 seconds.

Baseline Flatness

This test measures the instruments baseline flatness

Stability

This test measures the instruments stability. You can monitor the stability over one hour or half an
hour.

If you do not want to perform some of the tests you can undo the check boxes and only select the tests
you require. After you finish setting the validation parameters press the OK button and the system will
form a validation wizard according to your settings. You can now proceed with the instrument validation
by following the instructions as shown in Figure 3-7.

Figure 3-7 Instrument Validation Wizard

 21

If your instrument has a multiple cell holder you can select Auto Validation from the first page of the
validation wizard and setup the cell positions with the relevant reference materials. Then press Next and
the system will start the validation automatically. If you do not select the Auto Validation function you will
need to continue the validation procedure step by step. During the validation process the system will
indicate the tests which have failed. From the last page of the validation wizard, you can select the Print
Results Report button to print out the validation results. All tests which Pass are marked with a “” sign .

3.10 Chapter Summary

This chapter has given you an introduction on how to control an instrument using the UVWin5.0
functions.
 22

Chapter 4 Photometric Measurement

Key points
In this chapter you will learn how to do the following: What is photometric measurement? How to
setup parameters for photometric measurement? How to perform photometric measurements? How to save
and printout measured results?
It includes:
z A brief introduction to Photometric measurement
z Setting of photometric measurement parameters
z Photometric measurement
z Saving and printing measured results
z Chapter Summary

4.1 A brief introduction to Photometric measurement

The “Photometric measurement” is a simple readout of the measuring data at the wavelength points
you set. In UVWin5.0, you can set multiple wavelengths for photometric measurement. Furthermore you
can do some simple calculations on the measured data. Please refer to next section about this.

4.2 Setting of Photometric measurement parameters

To enable Photometric Measurement, you should click on the Photometry tab in the Work Space.
Select the Parameters settings submenu under Measure Menu to activate the Photometric measurement
setting dialog box, as shown in Figure 4-1. In this dialog box, there are 5 tabs and you can set them
according to your requirements.

Figure 4-1 Photometric Parameters Settings for (Fixed Wavelength Method)

 23

Measurement Tab

z Wavelengths
In the Measurement tab you can input the wavelengths you want to measure from the wavelength edit
box. First type the wavelength and then click the Add button to add it to the wavelength list below. You can
select up to 26 multiple wavelengths. If you need to delete any wavelength points or clear the wavelength
list you can press the Delete or Clear button. After you have chosen a wavelength the same wavelength
appears in the Wavelength Edit Box. Here you can amend the wavelength and press the Replace button to
update it in the Wavelength list.
z Repeat Measurement
The Photometric measurement application allows you to select a number of repeat measurements. If
you do not require repeat measurement select No in the Repeat Measurement Option. If manual repeat
measurement is required select Manual. Then input the number of repeat measurements in the Repeat
Times Edit Box, the same applies for a Manual Repeat.
The Automatic option also performs a repeat measurement. In the Automatic mode multiple repeat
measurements are made automatically without pressing the Measure Button. In the Automatic mode you
are required to set a time interval for measurement, this relates to the time between every two
measurements. This interval could be set to zero which means continuous measurements are performed
without any pause. If you have chosen Automatic Repeat you are also able to check the option box of
“Repeat measurement automatically according to cell number”, where one measurement is taken of all the
samples in the multi cell holder. In this case there is no need to set repeat times as this option will not be
available. If you set the cell holder type as a fixed cell holder then you cannot check this option box. The
main purpose for repeat measurements is for the averaging of repeat measurement data. So you can check
the “Calculate average value” to perform an average calculation. Then the system will calculate and
display the average automatically in the results table.
z Photometric Mode
Photometric mode refers to the current operating mode which has options of Abs. (Absorption), %T
(Transmittance), Es (Sample Energy), Er (Reference Energy), and %R (Reflectance).
z Start Number
Here you can set a start point for the sample serial number. This can be any chosen number of
preference.

Simple Calculation Tab

This option enables calculations to be performed on the measured results. Using this function, you can
apply simple formulae to work out customised analytical results. Its setting menu is shown in Figure 4-2.
You can check the option box for “Enable Simple Calculation” to open the simple calculation function.
 24

Figure 4-2 Photometric Settings (Simple Calculation Tab)

z Calculation formula

In the “Calculation formula” edit box you can input the required formula for results calculation. In the
formula letters A, B, C, and D…represent the measured data for corresponding wavelengths. For example,
if you input two wavelengths in to the Wavelength list of the Measurement Tab, these might be 600nm and
500nm. When you want to calculate the ratio of the measured data at these two wavelengths you can input
A/B in the Formula Edit Box of the Simple Calculation Tab. Then press the Add Button. The default title
of the calculation formula is “Result 1”, “Result2”… but you can specify a title at the same time when you
input your formula, in the Title Edit Box. If you need to amend a formula select it from the formula list,
and revise it in the formula edit box then press the Revise Button to make it effective. If you want to delete
or clear the contents of the formula list then press the Delete button or Clear button. You can have up to 10
different formulae.

z Characters
The role of the Characters here is for ease of input by imitating a keyboard. Press a character button
so as to input the corresponding character.

z Display
The role of the Display option here is for selecting different display modes for the calculation formula.
The pull-down box offers two choices, formula and title. If you select Formula that means the calculation
formula will be displayed in the results table as a title. If you select Title then the default title or user set
title will be displayed.

Instrument Tab

The Instrument Tab is the same as that for Instrument Performance Settings. Please refer to 3.1
Instrument Performance.

Accessories Tab

The Accessories Tab is the same as that for Accessory Settings. Please refer to 3.7 Accessory Settings.

Quality Control Tab

The Quality Control tab is a function which allows you to monitor the upper and lower limits of your
results. For example if you perform a measurement and get a value of 0.6, while its normal value should be
around 1.0 and absolute minimum should be not less than 0.8. Then it is clear that the measured result is
not right or is out of range. You can say its quality is not good as your criteria for passing is not less than
0.8.
 25

Figure 4-3 Quality Control Settings

The Quality Control is a new function offered by UVWin. Its role is quality monitoring on measured
data. If there are any abnormal data measurements the system will prompt you or act according to the
preferences you have set. As shown in Figure 4-3.

In the quality control window you can set the Quality Control function on or off through the “Enable
quality control function” option.
z Quality Control List
Here you can set the parameters up for quality control. The letters A, B, C and D… represent the
columns of measuring wavelength points, result1, result2, result3…, and the calculated results. Click the
All button to select all items, or if you click on the Invert Button the inverse selection will be made. Click
on Clear Button to clear all selections.
z Limitation (Limits)
In the limitation box you can input a High Limit and a Low Limit for the quality control of selected
items. A Fail Action can be set up for when the data measured falls outside the limits. The optional actions
include: Continue – continue to measure, Stop – stop measurement, and Remeasure – Measure the current
sample again. If you wish you can make a mark by the result which is outside of the limits in the result
table, click on the Marking button to set the mark type, as shown in Figure 4-4.

Figure 4-4 Quality Mark Marking Grid (Left) Marking Text (Right)
 26

Check the Enable Option to open the marking function. In the Mark box, you can select marking
mode for those data points outside of the limits. “Grid” is marking the data located in the grid. The
optional marks can be selected from a pull-down list in Grid. If you choose “Text” the color and font of the
text can be setup.
z Second fail action
Second fail action means that the system will take action when there are two in succession outside of
limits measurements. The options are “Continue” and “Stop”.

4.3 Photometric Measurement

To take a measurement in the Photometric Measurement mode just click on the button. The
measured result is displayed in the results table. If you want to delete a measured result, use the mouse to
click on it and select the Delete submenu under Edit Menu to delete it. If you need to recall the deleted
results you can click the right button of the mouse in the result table. From the popup menu select the
Cancel Deleting Submenu under the Delete Menu to recall the deleted results. If you want to hide the
deleted results uncheck the Display Deleted Sample under the Delete Menu.

4.4 Saving and Printout

Measured results can be saved to a file or printed out. After you finish making a measurement you can
select the Save submenu under File Menu, or click on button. The Save File Window will pop up, as
shown in Figure 4-5. Input the filename you want to save with and press the Save button to save files to a
specified location.

Figure 4-5 Save File Window

In the lower part of the window the name of the analyser is displayed. You can click on the Note
Button to make a note on the measured results. As shown in Figure 4-6.
 27

Figure 4-6 Note Window

You can input the sample name in the Sample Name edit box or input notes on the measured results in
the Comments edit box. All of the information above is saved with the results file for future reference.
If you want to print out the measured results you can select the Print submenu under File Menu, or
click on button. Naturally you must ensure that your PC has a printer installed, if not the system will
give an error prompt. In addition if you want to modify the printout format you can select Page Settings
submenu under File Menu, or click on button to open the Page Setting Window. As shown in Figure
4-7.

Figure 4-7 Page Settings Window

 28

Page Margin Tab

The Page Margin Tab allows you to set the parameters relating to margins. As shown in the above
figure. First you can set the units either Inches or Centimeters. Then you can input four margins of top,
bottom, left, and right in the Margin Box. This is the space between text and the paper edges. Also the
margin box is used to set the interval for the page header and page footer.

Format Tab

The Format Tab options are mainly used to set the format, font, table, and direction of page output. As
shown in fig 4-8.
z Text
Here you can set the text printing format. The optional formats include Normal, Simple, and Table.
“Normal” means printing out in text, without any decoration. “Simple” is to underline and highlight
headers. “Table” means printing out in a table. Also you can click on the Font Button to set the fonts.
z Row Spacing
Here you can set the interval of each text row. The optional row spacing includes “Minimum”, “0.5
Word”, “1 Word”, “1.5 Word”, and “Double Word”.
z Line Width
Here you can set the thickness of the table lines. The optional line widths include “Thin” and “Thick”.
z Date/Time Format
Here you can set the time and date format for the printout. The optional formats include “Weekday,
MM DD, YYYY HH:MM:SS”, “MM/DD/YY HH:MM:SS”, “MM/DD/YY”, and “HH:MM:SS”.
Except the first option, the year of date format is two digits.
z Page Number Format
Here you can set the page numbers for your printouts. The optional page number formats include “1,
2, 3…”, “-1-, -2-, -3-…”,”<1>, <2>, <3>…”, and “(1), (2), (3)…”. This means you can have a direct page
number printout, adding hyphens to both sides of the page number, adding angle brackets to both sides of
page number, and adding parentheses to both sides of the page number.
z Direction
Here you can set the orientation of your printout as Portrait (vertical print) or Landscape (horizontal
print).
 29

Figure 4-8 Format Tab

Page Header / Page Footer Tabs

The settings for the Header and Footer are the same. But the Header is printed at the top of the page,
whilst the Footer is printed at the bottom of the page. Also there are three positions to select for the Header
and Footer contents. As shown in Figure 4-9.

Figure 4-9 Header (Left) and Footer (right) Setting

z Left, Middle, Right

“Left”, “Middle”, and “Right” represent three parts of left, middle, and right for the header or footer.
You can select some options to print from the from pull-down menu. The optional contents include:
“None”: Not to print anything.
“Date/Time”: To print current date and time. The format of date and time can be set in Format Tab.
“Filename”: To print the filename of the measurement file.
“Module Name”: To print the name of current measurement module, such as Photometric, Spectrum
 30

Scan, etc.
“Instrument Name/Number”: To print the name and serial number of current instrument, such as
UV-Vis Spectrophotometer/01-1901-01-0001.
“Analyser”: To print the current login username.
“Company”: To print the current user’s company name.
“Page Number”: To print the current page number.
“Custom”: To print custom text. If you choose this option, there is a pull-down menu and from the
Edit Box you can input custom text.
Once you have input relevant contents, you can click on the “Font” Button to set its font. In addition,
you also can select the Separation Line to draw separation lines between the Header, Footer, and Text.

Figure 4-10 Print Parameters Setting

Print Parameters Tab

The Print Parameters Tab lets you select parameters relating to the measurements to be printed out.
The optional parameters include “Instrument Performance” and “Note”. The Instrument performance
includes the instrument name, serial number and bandwidth. The Note includes the analysers name, sample
names and a note.
If you want to see a preview of the final printout, you can select the Print Preview Submenu under
File Menu, or click on button to view a print preview. As shown in Figure 4-11.
 31

Figure 4-11 Print Preview

In the print preview window, click on or buttons to zoom the page. Click on button to
print. Click on button to perform page settings.

4.5 Chapter Summary

This chapter gives you an introduction of how to setup and perform measurements in the Photometric
Application. It also informs you about setting the options for printing and viewing a print preview,
functions which are common to other parts of the software.
 32

Chapter 5 Spectrum Scan

Key points
In this chapter you will learn about the following: What is Spectrum Scanning? How to setup the
method parameters for a Spectrum Scan? How to perform a Spectrum Scan? How to view spectrum
information? How to save and open spectrum files?
It includes:
z A brief introduction to Spectrum Scanning
z Setting the Spectrum Scan method parameters
z Spectrum Scanning
z Viewing spectrum information
z Saving and Opening spectrum files
z Chapter Summary

5.1 A brief introduction to Spectrum Scanning

Spectrum Scan is scanning a sample over a certain wavelength range with a defined wavelength
interval. A spectrum scan is a two dimensional display of Absorbance verses Wavelength. The Spectrum
scan is mainly used for the qualitative analysis of samples. It is also used to determine the maximum
absorbing wavelength for a sample so that a quantitative analysis can be performed. So it is a primary
application of UV-Vis Spectrophotometry.

5.2 Setting up the Spectrum Scan method parameters

If you want to perform a spectrum scan first you should set the scanning method parameters. To
enable the Spectrum Scan Window you should click on the Spectrum tab in Work Space. Select the
Parameters settings submenu under Measure Menu, or click on button, to activate the Spectrum scan
setting dialog box, as shown in Figure 5-1.
 33

Figure 5-1 Spectrum Scan Parameters Setting

In this dialog box, there are 3 tabs: Measurement, Instrument and Accessories. The Measurement Tab
is for setting the spectrum scanning method parameters, while the other two tabs are the same as those of
Instrument Performance and Setting Accessories. Please refer to 3.1 Instrument Performance and 3.7
Setting Accessories for the details.

Photometric Mode for Spectrum Scanning

Here you can set the Photometric Mode for Spectrum Scanning. The Photometric mode means
instrument current operating mode, which has options of Abs. (Absorption), %T (Transmittance), Es
(Sample Energy), Er (Reference Energy), and %R (Reflectance). If you choose the Energy mode (Er, Es)
you can click the Energy button to set the Energy Parameters.

Display Range

Here you can setup the Y-axis range for the spectrum scan. You can input relevant Maximum and
Minimum values from the Edit boxes.

Scanning Parameters

You can set the scanning method parameters for the wavelength range, wavelength interval, scan
speed etc. The Wavelength range consists of a Start and End. The Speed relates to the scanning speed. A
faster scanning speed will give comparatively less data quality, while a slower scanning speed will give
comparatively better data quality. The data interval is the scanning wavelength interval. This is the
wavelength interval in nanometers at which each data point is to be taken. The optional scanning intervals
are 0.1, 0.2, 0.5, 1.0, 2.0, and 5.0nm. The role of the Auto Sampling Interval is where the software chooses
a scanning interval for you automatically according to the scan range you have set.

Scanning Mode

The Scanning mode allows you to perform repeat scans of the same sample. Single scanning means
performing a single scan on the same sample. Repeat Scanning means performing multiple repetitive scans
of the same sample. Automatic Scanning is performing scans according to cell number and this depends on
the type of accessory you select. If you choose Repeat Scanning, you should set the Time Interval and
Repeating Times for scanning. If you choose Automatic Scanning you only need to set a time interval and
not the Repeat Times.
 34

Spectrum Scan

Select Start Submenu under Measure Menu, or click on button, to start a spectrum scan.
During a scan the system will show the live data being collected and displays a graph in the Spectrum Scan
Window. If you want to cancel scanning click on the Stop Button or press the ESC Key.

5.3 Viewing spectrum information

Select the Spectrum Information Submenu under the Graph Menu, or click on button, to open the
spectrum information windo, as shown in Figure 5-2. In the window all spectral items are displayed in the
list. You can click on any of them and its details will be displayed in the right window. You can also click
on the Colour button to change the display colour of the corresponding spectrum.

Figure 5-2 Spectrum Information Window

5.4 Saving and Opening Spectrum Scan Files

If you want to save a spectrum you can select the Save Submenu under File Menu or click on the
button to popup the Save Window. In the window you should input a filename and press the OK Button.
When you want to view the saved spectrum files you can select the Open Submenu under File Menu or
click on the Button to popup the Spectrum File Window, as shown in Figure 5-3.

Figure 5-3 Open Spectrum Files Window

 35

In this window you can select the spectrum that you want to open. On the right is a graph showing a
preview of the selected scan. Click on the Attribute Button to see the details of a spectrum file.

5.5 Chapter Summary

This chapter gives you information about spectrum scanning.

 36

Chapter 6 Quantitation

Key points
In this chapter you will learn about the following: What is quantitation? How to setup parameters for
quantitation? How to perform quantitative analysis? How to save measurement files?
It includes:
z A brief introduction to Quantitative Analysis
z Setting Quantitative method parameters
z Quantitation
z Saving measurement files
z Chapter Summary

6.1 A brief introduction to Quantitative Analysis

Quantitative Analysis is used to determine the unknown concentration of samples at a known

maximum absorbing wavelength. Standards of known sample concentration are used to set up a calibration
curve so that the concentration of unknowns can be identified. Quantitation has many measurement
methods, such as single wavelength, double wavelength, three wavelength, first order derivative, second
order derivative and third order derivative.

6.2 Setting up the Quantitative Method Parameters

To enable the Quantitation Window, you should click on the Quantitative tab in Work Space. Select
the Parameters setting submenu under Measure Menu, or click on button, to activate the Quantitation
setting dialog box, as shown in Figure 6-1.

Figure 6-1 Quantitation Parameters Setting

 37

In this dialog box, there are 5 tabs: Measure, Calibration Curve, Instrument, Accessories, and Quality
Control. Amongst them the Instrument and Accessory tabs are the same as those of Instrument
Performance and Accessory Settings. Please refer to 3.1 Instrument Performance and 3.7 Accessory
Settings for the details.

Measurement Tab

From the Measurement Tab you can setup quantitation methods, wavelength points, repeat
measurements etc. You can select different measurement methods for various determination requirements.
You can also set the sample number and measurement repeat times. As shown above.
z Measurement Method
Here you can setup the measurement method you want to use for quantitation. The options are: single
wavelength, double wavelength, double wavelength with coefficient, three wavelengths, first order
derivative (linear fit), second order derivative (quadratic fit), third order derivative, and fourth order
derivative. If you choose the single wavelength method you should input the measuring wavelength in the
Wavelength Edit Box. If you choose the double wavelength method or the three-wavelength method you
should input baseline wavelengths in Baseline Wavelength1 and Baseline Wavelength2 Edit Boxes. Please
refer to Appendix A Application Methods of Quantitation for details of Quantitation methods.
z Sample Number
Here you can set the Sample Identifier and Number for standards and samples.
z Others
In the Others Setting you can set duplicate measurements for standards and samples. Duplicate
measurement is performing multiple measurements on a standard or a sample. Then you can take the
average of the multiple measurement results into the content calculation. Therefore if you check the
Duplicate Measurement Option you are also required to input the necessary repeat measurement times, this
will take longer than the usual measurement time. If your instrument is equipped with a Multi-cell holder,
you can choose Cell Holder Interchange Automatic option. Then the system will move to the next cell after
it finishes one sample measurement, you need to place the standards and samples in order.

Calibration Curve Tab

The Calibration Curve Tab allows you to setup the parameters for generating calibration curves. The
optional parameters are: Curve Equation, Equation Order, Concentration Unit, Zero Interception, Curve
Evaluation, Calibration Method etc. As shown below.
 38

Figure 6-2 Calibration Curve Setting

z Curve Equation
Here you can select an equation to use for the calibration curve. There are two options: C=f(Abs) and
Abs=f(C). The former takes the absorption value as a variable to compute the concentration, whilst the
latter is an inverse function of the former mainly used for computing absorption coefficients.
z Equation Order
The Equation order relates to the order of the polynomial function of the calibration curve. The
options are: First, Second, Third, and Fourth order. The first order equation is called a linear equation,
whilst the others are called nonlinear equations. As UV-Vis Spectrophotometers have good photometric
accuracy and photometric repeatability the measured data has a linear correlation with respective
concentrations. So linear equations have a wider range of applications. You can select different equations
according to your actual situation, as shown in Figure 6-3 and Figure 6-4.

Figure 6-3 Linear Calibration Curve

 39

Figure 6-4 Nonlinear Calibration Curve

z Concentration Unit
Here you can setup the concentration unit for samples. The default concentration units have: ng/ul,
ng/ml, ug/ul, ug/ml, mg/l, ppb, ppm, and mol/l. Also you can input a new concentration unit manually.
z Zero Interception
Zero Interception adds a standard point of both zero values for concentration and absorption before
the first standard is measured. The zero point will be taken into account when fitting calculations.
z Curve Evaluation
Curve Evaluation is to evaluate the curve quality (correlation coefficient). The evaluation result is
displayed in the Preferences Window of the Quantitation Window. If you need to evaluate a calibration
curve you can set the Curve Evaluation Option as R2, Correlation Coefficient. Then click on the Setting
button to popup the Curve Evaluation Setting Window, as shown in Figure 6-5.

Figure 6-5 Calibration Curve Evaluation Setting

Here you can set the Evaluation Value and Positive Deviation as 1.0 and 0.0. As the correlation of the
calibration curve is expressed as a percentage it should take 100% as the evaluation value. Then the
correlation will never be larger than 100%, and the Positive Deviation will never be generated. So Positive
 40

Deviation is set as 0.0. Negative Deviation is generated on the basis of the evaluation value. For example,
if you set the Negative Deviation as 0.001, which means that your evaluation lower limit is
1.0-0.001=0.999. Your evaluation range is 1.0--0.999. The values out of this range are deemed as
unqualified data.
The Graphic Character is used to display the evaluation result. The Deviation Number relates to the
number of displayed characters. For example, if you set the evaluation value as 1.0, positive deviation as
1.5, negative deviation as 0.5, graphic character as “*”, and deviation number as 5. Then if you evaluate
the value of 0.5, you will get one “*”, but if you evaluate the value of 1.0, you will get five “*”. Because
you set a deviation number of 5, which indicates that the difference of positive deviation and evaluation
value, and the difference of negative deviation and evaluation value, are averagely divided into 5 sections
and each section is expressed with one “*”. As shown below.

0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5
* * * * * * * * * *

The evaluation procedure is as follows: first decide whether the evaluated value by comparing to
evaluation value is a positive or negative deviation. A value larger than the evaluation value is positive
deviation, whilst a smaller value than the evaluation value is negative deviation. After confirming it is
positive or negative the evaluation begins. The evaluation process is actually to see how close the
evaluated value is from the evaluation value. If it is closer then it gets more “*”. For example, by referring
to the above figure, the procedure for evaluating the value of 0.85 is: 0.85 is a positive deviation, so taking
0.5--1.0 as the evaluation range. First see if 0.85 is between 0.5 and 0.6, if not then add one “*”. Then see
if it is between 0.6 and 0.7, if not then add another “*”. It continues this same procedure until it gets to the
range of 0.8 to 0.9; here it ends because 0.85 is between 0.8 and 0.9. Thus there are a total of 4 “*”. This
procedure happens automatically without any exterior calculations. It is helpful for you to understand the
basic functions of curve evaluation, that is the more “*”, then the better the curve correlation is. These “*”
will be displayed in the Preferences Window of the Quantitation Window and can be printed out with
calibration curve.
z Calibration Method
The calibration method is divided into two types: Concentration Method and Coefficient Method. The
Concentration Method requires you to input concentrations of standard solutions. After the absorptions of
the standards are measured, the measured data is combined with the concentrations, and is fitted to
compute factors of equation and correlation.
The Coefficient Method requires you to input directly the factors for the equation, without measuring
the standards. For example if you set the equation order as second and select the coefficient method then
you need to input three factors of K0, K1, and K2. K0 is a constant, and K1 relates to the factor for the first
order and K2 relates to the factor for the second order.

Quality Control Tab

The Quality Control Tab option is the same as that for Photometric Measurement. Please refer to 4.2
Setting Photometric measurement parameters.
 41

6.3 Quantitation

The Quantitative Measurement Window is divided into 4 parts: Standards Table, Unknown Samples
Table, Calibration Curve, and Preferences. As shown in Figure 6-6.
Calibration Curve

Standards Table

Preferences

Samples Table

Figure 6-6 Quantitation Window

Measurement Table

The Standards Table or the Samples Table is used to display the measured results for standards or
samples. Generally the table holds 8 rows: Number, ID, Type, Concentration, Absorption, and Absorptions
at Wavelength, Standard Deviation, and RSD. If you choose the double wavelength method, or the three
wavelength method, the other wavelengths will also be displayed in the table.
z Number: Indicates the number of standards or samples.
z ID: This where you can type in the Standard and Sample names these can be edited.
z Type: For standards, the type is Standard. For unknown samples, the type is Unknown.
z Concentration: The concentration values of standards or samples.
z Absorption: The final absorption values of standards or samples. The calculation formula is
shown in Table 6-1.

Table 6-1 Quantitation Calculation Formula

Single Wavelength Method: Abs = A1

Double Wavelength Method: Abs = A1 − A2

Double Wavelength Coefficient Method: Abs = A1 − A2 • K

(W1 − W2 ) × ( A2 − A3 )
Three Wavelength Method: Abs = A1 − − A3
W2 − W3
 42

In the table, “Abs” relates to final absorption, “W1” relates to the first measuring wavelength also
called the main wavelength. “W2” is the second measuring wavelength also called baseline wavelength 1.
“W3” is the third measuring wavelength also called baseline wavelength 2. “A1” is the absorption at the
main wavelength. “A2” is the absorption at the baseline wavelength1. “A3” is the absorption at the baseline
wavelength 2. “K” expresses the coefficient of the Double Wavelength Coefficient Method.
z Absorption at wavelength: The absorption of standards or samples at corresponding wavelength
points.
z Standard Deviation: The Standard Deviation of repeat measurement.
z RSD: The RSD of repeat measurement. If repeat measurement is not checked, Standard
Deviation and RSD are both zero.
When you are ready to take standards or unknown sample readings select the corresponding
measurement table by clicking on it. Then at the top of the table it will display the word Actived
(ACTIVATED), which shows the table is enabled.

Calibration Curve

The Calibration Curve area is used to display a graph of the current calibration curve. If you do not
perform any standards measurements there is no graph displayed.

Preferences

Preference is used to display the current quantitation settings including curve equation and
coefficients.

Measurement

If you set the calibration method as the concentration method in the quantitation settings before
measurement you need to input your standards concentration values in the standards measurement table,
and only then will you be able to start taking readings. There are two ways to input concentrations. First
double click on the blank box of the ID Row, input an ID and press the Enter Key. The system will then
add a new standard column in the table automatically. Then click on the concentration box of the standard
and input its concentration. Or alternatively if you do not input any concentration values for your standards
but click the Start Button, this time the system will prompt you to input a concentration, as shown in
Figure 6-7.

Figure 6-7 System prompting input of concentration

After you have input the concentration, click on the OK Button. The system will add a standard
automatically with the concentration value you have entered and then it will start taking a reading of the
standard.
If you set calibration method as the coefficient method in the quantitation settings you do not need to
measure the standards. You can start the measurement of your unknown samples immediately.
 43

6.4 Saving measurement files

To save the quantitation data select the Save Submenu under File Menu or click on the button to
open the File Save Window. In the window input the filename you want to save to and click on the Save
button. If you want to add notes to the measurement file click on the Note button to input comments and
then press the OK Button.

6.5 Chapter Summary

This chapter gives you an introduction to the quantitative application module. Quantitation is an
important tool in UV-Vis Spectrophotometry and the correct application of its measurement methods can
add a valuable contribution to your analytical work.
 44

Chapter 7 Kinetics

Key points
In this chapter you will learn about the following: What is kinetics? How to setup method parameters
for kinetics? How to perform a kinetics measurement? How to save?
It includes:
z A brief introduction to Kinetics
z Setting Kinetic parameters
z Kinetics
z Saving time course curves to files
z Chapter Summary

7.1 A brief introduction to Kinetics

The Kinetics application part of the software allows you to monitor the rate of change of Absorbance
with time using a preset time interval and generates a graphical display of the measured data. The method
is mainly used for observation of sample change with time.

7.2 Setting Kinetic method parameters

To enable the Kinetics Window, you should click on the Kinetics tab in the Work Space. Select the
Parameters settings submenu under the Measure Menu, or click on the button to activate the Kinetics
setting dialog box as shown in Figure 7-1.

Figure 7-1 Kinetics Method Parameters

 45

In this dialog box, there are 3 tabs: Measurement, Instrument, and Accessories. The Instrument Tab
and Accessories Tab are the same as those of Instrument Performance and Accessories Setting. Please refer
to 3.1 Instrument Performance and 3.7 Setting Accessories for the details. The following will focus on
setting up the Kinetics method parameters.

Photometric Mode for Kinetics

Here you can setup the Photometric Mode for the rate analysis. You can specify the Photometric mode
to be Abs. (Absorption), %T (Transmittance), Es (Sample Energy), Er (Reference Energy), and %R
(Reflectance). The system will perform the measurement according to your selection.

Wavelength

Here you can enter the wavelength at which you wish to monitor the rate of the reaction. During the
Kinetics run the sample readings are continuously monitored at a predetermined wavelength so you should
specify the wavelength for collecting the data.

Display Range

Here you can set the Y-axis range for the Kinetics run. You can set the display range according to the
values generated by the samples.

Time Option

Time Option allows you set the measurement time for the kinetics run, the sampling number and the
time interval.
z Time Unit
Here you can set the minimum time unit for the Kinetics run. The options are: Minutes and Seconds.
For example, if you specify the time interval as 1 and the time unit as minutes then a reading will be taken
one time every one minute. If the time unit is set to seconds then a reading will be taken one time every
second.
z Display Mode for Time
If you specify the time unit as minutes you are able to select the display mode. If you select Regular
the system will display time in numerical format such as 50, 120, 300 etc. If you select Clock the system
will display the time in clock format like 00:00:50, 00:01:00, 00:05:00 etc.
z Automation Mode
In the Automation Mode the software will automatically select an appropriate sampling number and
time interval dependant on the Measuring Time. All you need is to specify is the Measuring Time. If you
do not check the Automation Mode then you need to specify the sampling number and the time interval.
z Measuring Time
The Measuring Time is the total time over which the rate will be monitored. This option is only
available when the Automation Mode is specified.
z Sampling Number
Sampling Number is the total number of readings to be taken during the Kinetics run. The sampling
number multiplied by the time interval is equivalent to the total time for the Kinetics run.
z Time Interval
Here you can set the time interval for the readings. It is also referred to as the Sampling Frequency.

Measurement Mode

The Kinetics application allows you to perform repeat measurements. You can select the
 46

Measurement Mode Tab to set the parameters for repeat measurement. As shown in Figure 7-2.

Figure 7-2 Setting Measurement Mode

z Single Measure
No repeat. Here the Kinetics run is performed just once.
z Repeat Measure
Here you can set the Repeat Times and Time Interval. The system will perform repeat measurements
according to your settings for the Repeat Times and each run is separated by the Time Interval. If your
instrument is equipped with an automatic multi-cell holder you can also select the Cell Holder Interchange
Automatic option. The system will automatically move to the next cell and then perform a kinetics run
before repeating the measurements again.
z Automatic Measure
If your instrument is equipped with an automatic multi-cell holder you can select the Automatic
Measure option. The role of automatic measure is to perform rate measurements on each cell. If you install
an automatic 8-cell holder the system will measure 8 time curves with automatic interchange of the cell
holder during measurement.

Time Increment

If you select the repeat measure or automatic measurement modes you can also set the Time
Increment. This allows you to add a time increment to the total measurement time.
z Start: It relates to the repeat time at which the time increment starts to be effective. That is the
time increment is added at the time that the repeat measurements have been performed.
z End: It relates to the repeat time at which the time increment is to be ineffective.
z Increment: This is the amount of time being added each time. The time is a percentage relative to
the last measure time.
 47

7.3 Kinetics

To start a kinetics run, you can select the Start Submenu under Measure Menu, or click on the

button to commence the measurement. During the measurement procedure the system will
display results graphically as shown in Figure 7-3.

Figure 7-3 Kinetics Window

7.4 Saving time course curves to files

Select the Save Submenu under File Menu, or click on Button to open File Save Window. You
can input the filename you want to save to and press the Save button to save it.

7.5 Chapter Summary

This chapter gives you an introduction to some of the key functions within the Kinetics Application
module. Kinetic analysis is the ability to monitor the rate of change of Absorbance or Concentration as a
function of time. This involves investigating the rate of increase and decrease of Absorbance as in Enzyme
reaction studies. Enzyme characteristics are studied in detail in clinical diagnostic laboratories and have an
important role in determining the extent of tissue damage in living organisms. When cells are damaged by
disease enzymes can leak into the bloodstream and the amount present can indicate the severity of the
tissue damage. Enzymes do not have absorption characteristics in the UV or visible spectrums but their
concentration can be analyzed by studying their catalytic properties. The rate or speed of a reaction can
determine the enzyme activity and thus their concentration.
 48

Chapter 8 DNA Protein Determination

Key points
In this chapter you will learn how to use the DNA protein determination function, how to setup the
method parameters, how to measure and how to process the measured results.

Contents include:
z Use of the DNA protein determination function
z Starting the DNA protein determination function
z Setting the measurement parameters
z Processing the result
z Chapter Summary

8.1 Use of the DNA protein determination function

As the structure of DNA and the secrets of gene expression are gradually exposed people have learnt
that genetic code is transcribed and expressed by RNA. Biologists are no longer satisfied with exploring
the secrets of biological inheritance they are more interested in studying the generic characteristics of
organisms at molecular level. When a generic code segment from the DNA of an organism is connected to
the DNA sequence of another kind of organism, a new generic material will be designed using human
intervention. This process is totally different to traditional methods of cultivating organisms to reproduce
offspring. The merging of a “gene” of one organism with the “gene” of another to create a new
combination of genes and create a new organism is similar to engineering design in biotechnology. The
biological technology of creating new organisms by reassembling genes is called “genetic engineering”.
Genetic engineering along with cell engineering, enzyme engineering, protein engineering and
microbiological engineering are all important branches of biological engineering.
Genetic analysis is a complex yet significant subject and genetic engineering has an important role to
play for the benefit of all human kind. The study of human genes has become an important scientific
research subject for developed countries. In genetic analysis the analysis tools play an important role,
therefore the UVWin 5.0 software includes the DNA/Protein determination function.
 49

8.2 Starting the DNA protein determination function

If you want to analyse DNA protein concentrations you can choose the “DNA protein analysis”
submenu under “application” menu, then the system will pop up a window for DNA protein determination.
As shown in figure 8-1, the window is divided into two parts. The central part is the display window
showing the current measured result. The bottom part is a control panel where you can start functions by
clicking corresponding buttons.

Figure 8-1 DNA protein determination window

8.3 Setting the measurement parameters

From the control panel there is a button named “setting”. You can open the parameter setting
window as shown in figure 8-2 by clicking this button.

Figure 8-2 DNA protein parameter setting

WL (A1, A2): represents the two measured wavelength of DNA protein.

Df (A1, A2): represents the two coefficients of DNA.
Pf (A1, A2): represents the two coefficients of protein.
 50

If you want to have 320nm background correction, you can click the “320nm background correction”
check box and input a relevant correction coefficient.

DNA protein can be calculated in the following way:

z Without 320nm correction
Density of DNA=Df1×A1－Df2×A2
Density of protein=Pf2×A2－Pf1×A1
z With 320nm correction
(A1=A1－A320, A2=A2－A320)
Density of DNA=Df1×A1－Df2×A2
Density of protein=Pf2×A2－Pf1×A1

8.4 Processing the results

The measurement of DNA protein concentration is quite simple. You simply put the samples into the
sample holder and then click the “measure” button on the control panel and the measurement is finished. If
you want to print the measured result click the “print” button on the control panel. If you want to export
the results you can click the “export” button.

8.5 Chapter Summary

This chapter discusses the methods used to determine the DNA and protein concentrations. You can
set the parameters according to your specific requirements.
 51

Chapter 9 Graph Tool Functions

Key points
In this chapter you will learn about the following: How to use the graph functions on a spectrum?
How to perform simple calculations? How to perform complicated calculations?
It includes:
z Use of the graph tool functions
z Analysis of spectra
z Creating of 3D spectrum
z Display settings
z Chapter Summary

9.1 Use of the Graph Tool Functions

With the development of computer technology IT has diversified into many fields. Manufacturers can
write improved professional processing software programmes to make analysis more convenient and time
saving. During the early development of UV-Vis spectrophotometers the first instruments processed data
by use of electrical devices such as recorders and plotting machines. Today’s high performance computers
can perform any graph process using powerful software such as UVWin.

9.2 Analysis of spectra

9.2.1 Display settings

Select the Setting Graph Parameters Submenu under Graph Menu to open the display settings window
as shown in Figure 9-1.
 52

Figure 9-1 Display Settings Window

z Display Gridding
Here you can setup the display of the vertical grid line. If you check the Display Gridding box you
can specify the type of line to display
z Display Scale Line
Here you can setup the type of scale lines to display for the coordinate axis. If you check the Display
Scale Line box you can specify whether the scale lines are drawn inside or outside of the coordinates frame.
The length of the scale lines can also be set.
z Automatic Separation
Here you can set the interval between the grid lines. If you check the Automatic Separation box the
system will distribute the grid lines automatically. If you want to specify an interval value, you can
uncheck the Automatic Separation box and set the Minimum Interval. The Minimum Interval is input as a
percentage value of the whole coordinate axis. If you set the Minimum Interval as 10% the system will
draw 9 grid lines to divide the whole coordinate axis into 10 sections. If you set the Minimum Interval as
20% the system will draw 4 grid lines to separate the whole coordinate axis into 5 sections.
z Color
Here you can set the color of the grid lines and scale lines.
z Central Color
Here you can set the color of the central graph.
z Frame Color
Here you can set the color of the frame of the graph displayed, coordinate values and its title.
z Preview
If you want to preview the graph you can check the Preview box and the system will display a live
preview of the graph whilst you are making changes to it.
z Default
Here you can restore the default settings.

9.2.2 General Observation

z Zoom Window
 53

You can Zoom in on the spectrum graph by selecting the Zoom Window submenu under the Graph
menu or by clicking on the button, the system will show it as a . Then you can move the mouse to
any position on the graph and press the mouse left key. Whilst holding down the left key drag the mouse to
draw a rectangle of the zoom area. Release the mouse to zoom in. To cancel the Zoom, select the Delete
Zoom submenu or click on button. You can also select the Coordinates Range submenu under the
Graph menu, or click on button to open the Coordinates Range Window, as shown in Figure 9-2. Here
you can set the coordinates range required and press the OK Button to display the required zoom effect.

Figure 9-2 Setting the Coordinates Range

z Measure Distance
Select the Measure Distance submenu under the Graph menu, or click on button. The system
cursor is shown as a . Then you can click the mouse left key at any position (P0) on the graph, then
move the mouse to another position (P1) and click the mouse left key again. The system will automatically
calculate the vertical distance and the horizontal distance between the two points P0 and P1, as shown in
Figure 9-3.

Figure 9-3 Distance Measuring Result

z Fit in Window
Select the Fit in Window submenu under the Graph menu, or click on button. The system will
automatically regulate the coordinate range of the spectrum graph and display all of the spectra normally.
z Peak-Picking
Peak-Picking detects all peaks and valleys on a spectrum and labels those peaks and valleys according
to the picking criteria. First open a scan spectrum from the file directory and then select the Peak-Picking
option from the submenu under the Graph menu click on the button. The system will automatically
detect peaks and valleys on the spectrum and show the results in a table, as shown in Figure 9-4.
 54

Peak mark

Valley mark

Results Box

Settings bar

Figure 9-4 Peak-Picking

After peak picking the spectrum window is divided into three parts: Spectrum window, Results Box,
and the Settings bar. The Spectrum window shows a mark for each picked peak and valley. The results box
displays the wavelengths, measured values and comments for each picked peak and valley. You can
improve the effect of the peak-picking by amending the parameters in the settings bar, as shown in Figure
9-5.

Figure 9-5 Settings bar for Peak-Picking

Threshold: Here you can set the minimum percentage limit of the height of picked peaks and valleys.
This is the percentage relative to the difference between the maximum value and the minimum value of a
spectrum. Peaks and valleys with a height larger than that of the threshold are picked out. You can input the
threshold in the Threshold Edit Box and press Return, or click on the ▼ button right of the edit box. In the
pull-down menu you can select the threshold.
Display: Here you can choose to display only the peaks or the valleys, or both of them.
Marks: Here you can set the type of mark for the picked peaks and valleys either as the value or as a
number. The mark value relates to the wavelength or the Abs of the peaks or valleys. The number is a serial
number for the peaks or valleys.
Direction: Here you can set the marking direction. The options are vertical or horizontal.
Frame: Here you can set whether marks have a frame or not.
Mark Note: Here you can choose the Comment Column details to be displayed, instead of the value or
number of the peaks and valleys. You can input a comment note for each of the peaks and valleys and then
check the Marking Note option.
 55

z Read Spectrum
The Read Spectrum tool uses a vertical line to scroll along the scanned spectrum and then gives a read
out of the data at the cross point of the vertical line with the spectrum. You can select the Read Spectrum
submenu under the Graph menu or click on the button. The system will draw a vertical line on the
spectrum. You can use the mouse to drag the line or use the keys ← and → to move the line and read out
the spectrum data.
z Read Screen
The Read Screen gives a read out of the data at the point where you place the mouse. You can select
the Read Screen submenu under the Graph menu, or click the button. The system will change the
cursor to a +. You can then move the cursor along the spectrum and the system will show the
corresponding coordinate values.
z Cross Hair Tool
The Cross Hair Tool is similar to the Read Spectrum Tool. The cursor is shown as a cross cursor.
Select the Cross Hair Tool from the Display Cursor submenu under the Graph menu, or click on the
button. The system will draw a cross cursor in the center of the spectrum. You can move the mouse to the
centre of the cursor and it will be displayed as a . Click the mouse left key to drag the cursor along the
spectrum. Click again to fix the cursor at the required position.
z Horizontal Stretch, Horizontal Shrink, Vertical Stretch, Vertical Shrink
Stretch and Shrink allows you to zoom in to your spectrum adjusting the coordinate axes separately.
You can select the Scale Zoom submenu under the Graph menu and choose the zoom action you require in
the submenu. Or you can click on the corresponding button on the toolbar: -- Horizontal Stretch,
-- Horizontal Shrink, -- Vertical Stretch, and -- Vertical Shrink.
z Curve Display
Curve Display offers you a variety of options for displaying your curve. The optional display modes
include: line, point, cross point, dot, and triangle point. You can select the Curve Display submenu under
the Graph menu and choose one of the display modes. Or you can click on the corresponding button on the
toolbar: -- line, -- point, -- cross point, -- dot, and -- triangle point.
z Combination Display
Combination Display allows you to display multiple spectra in a graph window. If your graph holds
over two spectra, you can select the Combination submenu under the Graph menu and choose the Build
Combination in the submenu, or click on the button to open the Combination Spectrum Window, as
shown in Figure 9-6.
From here you can select the spectra being combined and press the OK Button. If you want to delete a
combination display, you can select the Delete Combination next to the Build Combination, or select the
Delete All Combination option.
 56

Figure 9-6 Combination Window

z Display All Spectra

The role of the Display All Spectra is to display all of you spectra together in a graph. This allows you
to compare all spectra in one area. Select the Display All Spectra submenu under the Graph, or click on the
button.

9.2.3 Mathematic Calculations

The Mathematic Calculations you will find here are commonly used in spectral analysis. When
qualitative analysis is required for those samples with baseline interferences the Mathematic Calculations
play an important role. In UVWin5.0, there are three kinds of calculation functions: basic, advanced and
additional.

Basic Calculations

The basic calculations include Addition, Subtraction, Multiplication, and Division. You can select the
Addition submenu, the Subtraction submenu, the Multiplication submenu and the Division submenu under
the Mathematic Calculation menu, as shown in Figure 9-7.

Figure 9-7 Basic Maths Calculations

z Source: Choose the spectrum on which the calculations are to be performed.

z Calculation: Choosing the operation you want to perform
z Spectrum: Here you can input a value which relates to a constant being applied or you can
choose a spectrum.

Then press the OK button and the system will display the new curve with the results of the
calculation.

Advanced Calculations

The Advanced Calculations provide the option to smooth the data or to convert to First Order
Derivative, Second Order Derivative, Third Order Derivative and Fourth Order Derivative. You can select
the Smooth submenu or the Differential submenu under the Mathematic Calculations menu. The system
will open the Math Calculation Setting Window, as shown in Figure 9-8.
 57

Figure 9-8 Advanced Math Calculation

z Spectrum: Select the spectrum to be smoothed or derivatised.

z Calculation: Choosing the calculation you require.
z Number of Points: You can select the number of points for smoothing or for the derivative
operation. The smoothing algorithm uses the number of data points to be averaged to give a
result.
z Coefficient: Here you can set a multiplier value for magnification of the calculated results. If you
set this to 1 then there is no magnification, a value of less than 1 expresses reduction and a value
bigger than 1 expresses magnification.

Additive Calculations

The Additive Calculations allow you to convert the spectrum to Log10, Reciprocal and to transform
the spectrum from Abs to T%. You can choose the Logarithm submenu, the Reciprocal submenu, and the
Transform submenu under the Mathematic Calculation menu. The system will open a window, as shown in
Figure 9-9.

Figure 9-9 Additive Mathematic Calculation

z Spectrum: Selecting the spectrum to be converted.

z Calculation: Choose the calculation method.
 58

9.2.4 3D Spectra

3D Spectra is a new feature in the UVWin Software. Its role is to combine multiple spectra into a 3D
display. The 3D application allows you to compare multiple spectra in three dimensions this is a useful tool
for qualitative analysis and can also be used when monitoring reaction rates using the Kinetics application.
Once you have scanned several spectra or have recalled spectra from file, you can select the Establish 3D

spectrum submenu under the Graph menu, or click on the button to open the 3D Spectra Window, as

shown in Figure 9-10.

Menu and Toolbar

Spectra Tree

Graph Display

Cascade Display
Figure 9-10 3D Spectra

The 3D spectra window is divided into 4 parts: Menu and Toolbar, Spectra Tree, Color Cascade
display and Graph Display.

Menu and Toolbar

The Menu and Toolbar provide most functions of the 3D Spectra application. Here is a description of
the function of each menu item.
File:
z Save as bitmap: Saving 3D spectra as a bitmap file.
z Copy to Clipboard: Copying 3D spectra to the system’s clipboard, so that you can cut and
paste from the clipboard to another package.
z Print: Printing 3D spectra out. Since 3D spectra contain various colours it is impossible for
a black and white printer to distinguish the different colours. Therefore it is recommended
that you use a colour printer for 3D spectra printout.
z Exit: Quitting the 3D spectra window.
 59

Graph:
z Original Viewing Angle: You can reset the viewing angle.
z Perspective Projection: Here you can set the projection mode to perspective.
z Orthographic Projection: Here you can set projection mode to orthographic.

Tips: “Projection Mode”

The “Projection Mode” is an important concept for 3D systems. Its role is to make a 3D graph
look real. For example in the real world, the objects you see near to you look larger than those objects
far away from you. This is due to the angle of viewing. Therefore 3D systems use different projection
modes to achieve the best viewing angle to imitate the real world. Projection Modes include
Perspective Projection and Orthographic Projection. The Perspective Projection performs a projection
which imitates the real angle of view of the actual world. Perspective Projection is commonly used in
3D scene design software such as 3DSMAX and House design software etc. Orthographic Projection
does not imitate the real angle of view but it shows the graph directly. The graph displayed with the
Orthographic Projection cannot be changed irrespective of objects being near and far away.
Orthographic Projection is mostly used in CAD software development.

z Zoom in: Magnifying a 3D graph.

z Zoom out: Minimising a 3D graph.
z Vertical rotation: Performing rotation round the X axis.
z Horizontal rotation: Performing rotation round the Y axis.
z Show mini-coordinate axis: Displays the mini-coordinate axes in the lower left corner of the
window.
z Show spectrum coordinate axis: Displays the coordinate axes and its gridding.
z Display spectral curves: Displays 3D spectra as curves.
z Display spectral surface: Displays 3D spectra as surface.
z Display spectral wire frame: Displays 3D spectra as a wire frame.
z Illumination effect: Adds an illumination effect to the 3D spectra. It is only in the spectra
surface mode that the illumination effect can be observed.
z Front view: Adjust the visual angle to front view position.
z Top view: Adjust the visual angle to top view position.
z Side view: Adjust the visual angle to side view position.
Colour
z Coordinates colour: You can set the colour of the coordinate axes.
z Character colour: You can set the colour of the text in spectral graph.
z Background colour: You can set the background colour of the 3D spectral graph.

Spectra Tree

The Spectra tree is used for displaying the names of the current spectra. You can display or hide
spectra by selecting the check boxes infront of the spectral headers.

Peak-Picking

If you want to perform peak-picking on 3D spectra, select the Peak-Picking option, and then input a
detection threshold in the Edit Box and press the Pick button. The Peaks are detected automatically and
displayed in the results peak-picking table, as shown in Figure 9-11. If you want to close the peak-picking,
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check the Close Display option.

Figure 9-11 Peak-Picking of 3D Spectra

Layering Display

The Layering display allows you to separate 3D spectra into several layers by using different colors so
that you can observe any changes in spectral trends more clearly. Here the 3D facility offers five layers and
you can set up the colours according to your requirement. As shown in fig 9-12.

Figure 9-12 Layers Setting

In the layers setting window you can set each layer to on or off. Also you are able to specify a value
range and colour for each layer. Each layer holds an upper and lower limit and a colour association. For
example the first layer in the above figure holds a lower limit as 0.000, an upper limit as 20.0000, and a
color as red. It states that in the 3D spectral graph, any number of spectra larger than 0 and smaller than 20
are considered as the first layer and are displayed in red. The second layer, which is located above the first
layer, has its lower limit as the upper limit of the first layer and its upper limit is 40.000, its colour is
yellow. This states that any number of spectra greater than 20 and smaller than 40 are considered as the
second layer and are displayed in yellow. The same condition can be applied up to the fifth layer. If you
want to set the colour of each layer then click on the color tab. The bottom colour tab relates to the number
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of scans that are smaller than the lower limit of the first layer, whilst the top color tab relates to the number
of scans that are bigger than the limit of the fifth layer.
Layering not only can be lengthwise, but also transverse. You can choose the Vertical button or the
Horizontal button for the Layering direction. In addition click on the Reversion button to reverse the order
of the colour tabs.

Graph Display Window

The Graph Display Window displays all of the 3D spectra and allows you to perform a few simple
operations on the spectra with a mouse. You can hold down and press the left button of the mouse and
move the mouse to make the 3D spectra rotate. Similarly if you hold down and press the right button of the
mouse you can move the mouse to zoom in and zoom out of the 3D spectra.

9.3 Chapter Summary

This chapter gives you a detailed introduction to the graph tool bar operations in UVWin 5.0. The
graph tools available can be very helpful to those users requiring added functionality and flexibility for
manipulating and displaying results in a professional way.
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Chapter 10 Exporting Data

Key points
In this chapter you will learn about the following: What is Data Export? What kind of data can be
exported? How to export data? How to handle the exported data files?
It includes:
z A brief introduction to Exporting Data
z Exportable Data
z Exporting Data
z Handling exported data files
z Chapter Summary

10.1 A brief introduction to Exporting Data

The main goal of any chemical analysis or scientific research is to produce quality data. Sharing data
has become an integral trend in today’s society and the scientific community. For example, a results table
prepared with MS Excel can be inserted into MS Word for the very purpose of sharing data. Sometimes
you might also need to import your measured data into another editing software package. UVWin5.0
software allows you to export data to other editing software packages such as MS Excel and MS Word by
using the Export function which is based on Windows OLE. This gives you enhanced capabilities, more
flexibility and makes your analysis simple and convenient.

Tips: “OLE”
OLE is the abbreviation for Object linking and embedding. It is a standard interface for programming
supplied by the Windows Operating System. An application program can use the interface to access those
applications that offer OLE functionality. MS Word and MS Excel supply the OLE function and so other
applications can use them through OLE.

10.2 Exportable Data

In UVWin5.0 the data which is capable of being exported is mainly the measured data. You can also
export the instrument information. To do this you can select the last submenu of Information under the
Measure Menu, as shown in figure 10-1. In the window it shows the information about current instrument
hardware and includes: Model, Serial Number, Wavelength Range and Spectral Bandwidth. In the lower
left corner of the window there is an Export button. Pressing the button will export the information into the
relevant files.
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Export Button

Figure 10-1 Exporting instrument information

10.3 Exporting Data

Exporting data is easy and simple. You need to select the Export Data Submenu under the File Menu.
The system will open the Data Export Window, as shown in Figure 10-2. Here you can specify the file type
for export by choosing the Export Type option (please refer to Table 10-1 for exportable file types). In the
edit box of Export File input the filename being exported, or click on the “…” button to the right of it to
specify a file. Then click the Export button. The system will export all the data to the appointed file
according to your settings. If you check the option box of “Open files automatically after Export”, the
system will open the exported file automatically once the exporting has completed.

Figure 10-2 Data Export Window

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Table 10-1 Exportable File Types and Descriptions

File Type Description Format Remark

Microsoft Word Documents Row 1: Title Title Title… Microsoft
Word File Extension: “.doc” Row 2: Data Data Data… Word is
… required.
Microsoft Excel Worksheet Row n: Data Data Data… Microsoft
Excel File Extension: “.xls” Excel is
required
The Text File of comma (,) Row 1: Title, Title, Title…
Text File
delimited between each data Row 2: Data, Data, Data…
(Comma
point which can be opened …
Separated
with Excel. Row n: Data, Data, Data…
values)
Extension: “.csv”
The Text File of Tab delimited Row 1: Title<Tab>Title<Tab>Title…
between each datum. Row 2: Data<Tab>Data<Tab>Data…
Text File
Extension: “.txt” …
Row n: Data<Tab>Data<Tab>Data…
A non-delimited ASCII (ASC) Row 1: Data 1
file Row 2: Data 2
ASC File Extension: “.asc” …
Row n: Data n

10.4 Handling exported data files

Once the data has been exported you have the flexibility to do whatever you want. Also if you need to
compose a research paper and you think that the data reports printed out with UVWin5.0 do not meet your
requirements you can export the data to a Word file. Then you can edit the word file as you require. Also if
you want to perform more complicated calculations you can export your data to Excel and then use the
formulae in Excel to perform the calculations on the data.

10.5 Chapter Summary

This chapter gives you an overview of the Export function within UVWin 5.0.
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Chapter 11 Administration

Key points
In this chapter you will learn about the following: Why the Administration Facility is an important
part of the software? How to setup users and groups? How to set security parameters? How to setup and
browse the operation log file?
It includes:
z A brief introduction to the Administration Facility
z How to create a user account
z How to create a user group
z How to setup the security parameters
z How to setup and browse the operation log file
z Chapter Summary

11.1 A brief introduction to the Administration Functions

Users of Microsoft Windows will know that since Win2000 you have been able to setup different
users to log on to the system, where you can use the Administrator to create users with different privileges.
The same principles apply to the Administration functions within the UVWin Software. The
Administration Facility is to allow multiple users to have different levels of accessibility to the software
using passwords set up by the main administrator. UVWin5.0 includes Multiple User Management,
Security System and Log Management.

11.2 Managing Users

User Management is a basic administration function in UVWin5.0. This function will assist you in
creating users and groups and setting relevant parameters. You can select the User Group submenu under
the Administrate menu to open the Management Window. In the window, there are two tabs: Users Tab and
Groups Tab. You can create users and change user’s settings in the User Tab, and create groups and modify
group settings in the Group Tab, as shown in Figure 11-1.
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Figure 11-1 User Management (Left) and Group Management (Right)

11.2.1 User Management

If you select the User Tab in the Management Window you are able to manage the users. You can
create a new user and delete or modify each user’s settings.
z Create
The function of New allows you to create a new user. Click on the New button and the system will
pop up the New User Window, as shown in Figure 11-2. In the window you can enter a user name in the
User Name Box and specify a group for the new user in the pull-down list of the Group Box. The role of a
group is to manage various users by grouping and setting different privileges. For details on groups, please
refer to the next section.

Figure 11-2 New User Window

If you need to create a password for a new user, click on the Amend Password button. The system will
open the Password Window, as shown in Figure 11-3. You need only to enter the password you want in the
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Password Box and then again in the Confirm Password Box and then click OK. If you want to prompt the
user to change their password the next time they sign in, you can check the option box User Should Amend
Password at Next Sign-In. If you want to terminate the user account, check the option box Account is
terminated.

Figure 11-3 Enter Password Window

In addition you can also input any relevant information for a new user in the Information boxes. The
information includes: Full Name, Company, Section, and Description.
After you finish setting up a new user click on the Create button. The system will create a new user
account according to your settings. Then you can log on with this user account by entering the user name
and its password at your next sign-in.
z Rename
Here you can change the name of the current user. From the user list specify the user being renamed
and click on the Rename button. The system will prompt you to input a new username, as shown in Figure
11-4. After you enter a new name, click OK to return.

Figure 11-4 Rename a Username

z Remove
The Remove function allows you to delete a specified user. In the user list select the user to be deleted
and click on the Remove Button. The system will prompt you to confirm the deletion. Select OK to delete
the user. If the user you select is the current log on user, the system will prompt you with a message
“unable to delete”. In this case you need to use other users to log on to delete this user.
z Copy
The Copy function creates a copy of the current specified user. Apart from the username other settings
are the same as the user you specify.
z Property
The Property function allows you to view user information. Click on the Property button, and the
system will open the User Attribute Window. The window is similar to the one for creating a user. In this
window you can modify the relevant information for the user you have selected.
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11.2.2 Group Management

From the Group Tab in the Management Window you are able to manage the groups. Here you can
create a new group and delete or modify each group’s settings.
z Create
The New button allows you to create a new group. Click the New button and the system will pop up
the New Group Window, as shown in Figure 11-5. You can enter a group name in the Group Name Box
and input the description information for the new group in the Description Box.

Figure 11-5 New Group Window

From here you can set up the group’s privileges. You can setup a list of permissions for a group
according to its requirements. These privileges will apply to each user within the group. If a user is part of
the group then when the user logs on their accessibility rights will be limited to the permissions allocated
to that group.
z Rename
The Rename function allows you to change the name of the current group. In the group list select the
group being renamed and click on the Rename button. The system will prompt you to input a group
username. After you enter a new name, click OK to return.
z Remove
The Remove Function allows you to delete a specified group. From the group list select the group to
be deleted and click the Remove button. The system will prompt you to confirm the deletion. Select OK to
delete the group you have selected. If the group you specify includes the current log on user the system
will prompt you with the message “unable to delete”. In this case you need to use another group user to log
on to delete this group.
z Copy
The Copy Function will create a copy of the current specified group. Apart from the group name other
settings are the same as the group you specify.
z Property
The Property Function allows you to view the group information. Click on the Property button and the
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system will open the Group Attribute Window. The window is similar to that of creating a group. In the
window you can modify the relevant information for a group.

11.3 Security

The security features within the UVWin5.0 Software guarantee the user peace of mind that data integrity is
not compromised and that the software offers a safe and secure operating environment. You can achieve
data protection by selecting the Safety Settings submenu under the Administrate menu to open the Security
Window, as shown in Figure 11-6.

Figure 11-6 Security Settings

Password Valid Period

Just as its name implies here you can set the time for the Password to be valid. This is how long a user
password can be used for. If you want to use a password over an unlimited time period then you can select
the “Never Expired” option. If you want a user to change their password every 30 days then you should
choose the other option and enter the required number of days in its edit box.

Password Length

The password length is used for setting the length of a user password. You can specify the maximum
length and the minimum length. In addition if you do not specify the minimum length then a blank
password is allowed to be selected. In this case a user can use a blank password which is no password.

Security

The function of the “Auto Login by Administrator” is to start the UVWin5.0 software without
displaying the login window. The system will use the administrator account to log in. For a single system
user it might be more convenient for them to use the software without entering a password to log in.
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If you do not check the option, it means you need to enter an account and its password for each login.
Then you are able to set the times for failed loggings. That is the system will count the number of failed
attempts to login, once this reaches the number you set the system will exit you automatically.
If you wish to lock the UVWin Software after being idle for a period of time then check the Lock
System option and select the time for an automatic lock to take place.

File Protection

The function for file protection is to protect your data files. This means that data files created by users
cannot be read by other users. Only the creator or administrator can read and open a data file.

11.4 Operation Log

The log function is to record all actions that take place within the software. You can select the View
Log submenu under the Administrate menu to open the Operation Log Window, as shown in Figure 11-7.

Figure 11-7 Operation Log Window

You can select which log to view from the left window and the details of that log are displayed in the
right window. If you want to set the log parameters you can select the Option submenu under the Operation
menu, the system will open the Log Option Window, as shown in Figure 11-8.

Figure 11-8 Log Option Window

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Save in: Here you can set the set the path name for the Log File. For example you could save the Log
File to another hard disk to prevent data being lost or as a backup.
Maximum Record Number: Here you can set the maximum record number allowed for a Log File to
save. When the record number is reached the system will act according to your choice.

11.5 Other Administrative Functions

11.5.1 User-Defined Log

If you want to input a user-defined log record, you can select the Record Log submenu under the
Administrate menu. The system will open an input window. Enter what you want to record and click OK.
The system will record any actions in the Log database.

11.5.2 Lock System

As mentioned above you can lock the system by using the”Lock System after Idle” in Security
Settings. If you want to lock the system at any time you can select the Lock System submenu under the
Administrate menu or press the Combo Keys Ctrl+Alt+Space, to lock the system. After the system is
locked you will be required to enter the correct password to release the lock. As shown in Figure 11-9.

Figure 11-9 System is locked

Only the current user or the administrator can release the status of the system lock. If you enter the
wrong password the system will be unable to release the lock. If you choose Cancel the system will exit
automatically.

11.6 Chapter Summary

This chapter gives a detailed introduction to the Administration Application in UVWin 5.0.
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Chapter 12 Conclusion
The information provided in this manual gives you a detailed overview of the main functionality of
the UVWin 5.0 Software and if you have had time to use this manual you will have gained a good
understanding of how to get the most out of the software.
Users will gain confidence in using the software and practising some of the procedures by referring to
this manual on a regular basis.