Good Laboratory Practise and Quality

Assurance of Photometer and Equipment

Adam Aradea
Application Specialist for WFA & LW
MerckMillipore Division, PT Merck Tbk
GLP and other quality systems

Several quality systems exist worldwide

The following are referring to laboratory

testing and analysis:

ISO 17025
GMP (Good Manufacturing Practice)
GLP (Good Laboratory Practice)
GCP (Good Clinical Practice)

GLP and GMP show many similarities when

compared concerning physical-chemical tests.

2 WFA Professional quality assurance | 2012

Quality assurance in
photometric water analysis

Water analysis plays a key role in the

monitoring of operating facilities

Defined basic conditions are provided in:

Quality Assurance
regularily
qualified
documented
affordable effort

3 WFA Professional quality assurance | 2012

Definitions

Analytical Quality Assurance (AQA)

Measures for supporting own research results under controlled conditions.
Analytical quality assurance comprises internal and external quality control.
(according to DWA-A 704, 2007)

Laboratorium harus mempunyai prosedur pengendalian mutu untuk

memantau keabsahan pengujian dan kalibrasi yang dilakukan.
(according to SNI ISO/IEC 17025:2008 - 5.9.1)

4 WFA Professional quality assurance | 2012

Definitions according to DWA-A 704, 2007

Internal Quality Control (IQC)

Measures for quality certification that are taken by the user imself, when
using operation methods.
External quality control
Measures for quality certification, supported by external experts/laboratories
in case of application of operating methods.
Interlaboratory tests (round robins)
Instrument of external quality assurance. Analysis of split samples by
different laboratories.
Testing equipment
Equipment and reagents used in analyses.

5 WFA Professional quality assurance | 2012

Testing Equipment Monitoring

Testing equipment: All devices and reagents that are used in operating analysis
Testing equipment monitoring plays a major role in quality assurance
All laboratory equipment with essential influence on the measurement result has to be
tested
For a proper testing equipment monitoring, a list of equipment is required

1 2 3 4
Example:
Equipment list Testing equipment Assessment Measure Date
Treatment plant Hoppenstdt signature
pH-electrode documented in see there 04 January 2004
equipment log Peters
Oxygen measuring o.k. none 04 January 2004
device Peters
Photometer maintenance by protocol see 16 February 2004
customer service; ok maintenance Franu
documents

6 WFA Professional Quality Assurance | 2012 Source: Norm DWA-A 704 / State 2007, IQC-Card 9, Sheet 1
Recommendations for control and monitoring
frequencies
DPLP 07 SR03 JAMINAN MUTU PERALATAN YANG DIGUNAKAN OLEH
LABORATORIUM PENGUJIAN KIMIA DAN BIOLOGI

Testing equipment Monitoring

Photometer quarterly
Heating device/ quarterly
thermoblock
Thermometer Half-yearly

Scales quarterly

7 Source: Norm DWA-A 704 / State 2007

Recommendations for control and monitoring
frequencies according to DWA A 704

Testing equipment Monitoring

Photometer annually
Heating device/ annually
thermoblock
Piston stroke pipettes quaterly

Sampling equipment biannually

Reagents quaterly
Thermometer quaterly
Scales annually

8 Source: Norm DWA-A 704 / State 2007

Internal Specifications
If there are special working regulations for the testing equipment, they can be
listed in column 4 of Sheet 3.
Especially the recommendations made by the manufacturers must be
followed.
Example: Wastewater system: Treatment plant Hoppenstdt
1 2 3 4 5 6 7 8

Testing equipment Description Device no. Testing Frequency of Quality target stocked on rejected on
(device) (brandt) (if required regulation (if checking
location) available)

Photometer Lambert LE 5 IL * 1 TR 296 1 X per year Maintenance. by cust. serv. ok 13 Oct 2003

Thermoblock Brutzel 25 IL * 2 TR 27 1 X per year <= 3 C 13 Oct 2003

pH-meter Ionomess AB IL * 3 TR 135 1 X per week <= 0.2 pH 13 Oct 2003

Conductivity measuring device Messimax 4 IL * 4 TR 136 4 X per year <= 5 % for 500 S/cm 13 Oct 2003

BOD measuring device Respiroprompt IL * 5 TR 165 2 X per month <= 10 % calibration table 13 Oct 2003

Oxygene measuring device Oxistar OX IL * 6 TR 168 4 X per year <= 10 % 13 Oct 2003

Pipette 200 L Mikromess 200 IL * 7 TR 13 4 X per year <= 2 % 13 Oct 2003 15 July 2004

Pipette 500 L Mikromess 500 IL * 8 TR 13 4 X per year <= 2 % 13 Oct 2003

Pipette 1000 L Mikromess 1000 IL * 9 TR 13 4 X per year <= 1 % 13 Oct 2003

Pipette 2.00 mL Mikromess 2000 IL * 10 TR 13 4 X per year <= 1 % 13 Oct 2003

Pipette 200 L Mikromess 200 IL *11 TR13 4 X per year <= 2 % 17 July 2004

9 Source: Norm DWA-A 704 / State 2007, IQC-Card 2, Sheet 3

Checking the pipettes
Verification of piston stroke pipettes:
DWA A704
Standard procedure: Checking with a balance
Quality assurance by taking a volume control
Aim: Validation of accuracy
Recommended minimum frequency for validation: 4x per year

Quality target:
For pipettes with volume 100-1000l Tolerance < 2%
For pipettes > 1000l Tolerance < 1%

11
DWA A704 procedure

Pipettes with variable volume should be checked with three different

volumes (10 %, 50 % and 100 % of the maximum volume).
If the results lie outside the tolerance range, the pipette must no longer
be used as testing equipment (remove designation) and should be sent to
the manufacturer for testing.

12 Source: Norm DWA-A 704 / State 2007

Interpretation and documentation of the
measuring results
The results should be written down on a protocol sheet.
Five weightings with one pipette can be done. The results
could have small deviations. The calculation of a mean value
from these five weightings give a statement about the accuracy
of the pipette.
The mean value must lie within a defined tolerance range (see
next page).

14
 Example: Testing Equipment Monitoring
Pipettes
Wastewater system: Treatment plant Hoppenstdt
1 2 3 4 5 6 7 8
Pipette No.: 1 1 1 2 2 2 3
Maximum volume in ml 5.000 5.000 5.000 1.000 1.000 1.000 1.000

Variable? Y Y Y Y Y Y N
Remark Test Test Test Test Test Test
10, 50, 100 % 10, 50, 100 % 10, 50, 100 % 10, 50, 100 % 10, 50, 100 % 10, 50, 100 %
of max. volume of max. volume of max. volume of max. volume of max. volume of max. volume
Test volume in ml 0.500 2.500 5.000 0.500 1.000 1.000 1.000

Measured value 1 in g 0.518 2.552 4.963 0.505 0.905 1.005 1.005

Measured value 2 in g 0.512 2.450 4.964 0.495 0.884 1.033 1.013
Measured value 3 in g 0.520 2.605 4.995 0.488 0.899 0.993 1.002
Measured value 4 in g 0.490 2.551 5.033 0.493 0.911 0.922 1.012
Measured value 5 in g 0.505 2.533 4.895 0.496 0.893 0.996 0.996

Mean value in g 0.509 2.538 4.974 0.495 0.898 1.004 1.006

Deviation to test 1.8 1.528 -0.52 -0,92 -10,16 0,42 0,600

volume in %
Quality target in % 2 2 2 1 1 1 1

Quality target yes yes yes yes No! Pipette Yes yes
achieved? adjusted wrongly Repetition
Date 11 July 2004 11 July 2004 11 Juni 2004 13 August 2004 13August2004 13 August 2004 15 Sept. 2004
Signature Bast Bast Bast Klumm Klumm Klumm Bast

15 Source: Norm DWA-A 704 / State 2007, IQC-Card 9, Sheet 3

Standard method vs. Spectroquant
PipeCheck
Standard method Spectroquant PipeCheck
Check of pipettes according to DWA A704

very time-consuming very easy and fast verification by

absorbance values
highly precise and regularly
calibrated balance is needed no balance necessary
mobile application
Shelf life of PipeCheck is 3 years
Test solutions (4x6 cuvettes) =
sufficient for 6 tests of four different
volumes
4 reference solutions included

16
Monitoring of pipettes with Spectroquant

Spectroquant PipeCheck, 1.14962.0001

The respective check solution is diluted with distilled water using the
pipette to be checked.
The absorbance of the diluted solution is compared with that of the
included reference solutions.
Pipettes exhibiting deviations in volume exceeding 2.5 % are rejected as
defective.

17
 Spectroquant PipeCheck

Example: Calculation of upper and lower limit

in the package insert of the PipeCheck.

"Check result within the range of tolerance

of the reference result?

Reference solution included in the box

Check solution: Pipette into a "Check" cell with

18 the pipette to be checked and mix
Monitoring of cells
and reagents
Monitoring of cuvettes

Cells must be clean and scratch-free

Round cells at cell tests:
- Round cells are used only once No contamination and scratches

Rectangular cells:
- Clean after using
- Residues of purifying agents can influence the test result
- Two cells in the same package ("matched cells"), cells are adjusted and have similar
absorbance
- Insert rectangular cells always in the same direction into the photometer

Plastic cells:
- Different absorbance characteristics

20
Cells from Merck

Rectangular cells 10mm (Ord. no. 1.14946.0001)

Rectangular cells 20mm (Ord. no. 1.14947.0001)
Rectangular cells 50mm (Ord. no. 1.14944.0001)
Rectangular quartz cells 10mm (Ord. no. 1.00784.0001)

Semi-micro cells 50mm (Ord. no. 1.73502.0001)

Round cells 16mm (Ord. no. 1.14724.0001)

21
Control of the shelf life of the reagents

DWA A704 recommends:

Check the shelf life of the reagents quarterly

Close the reagent bottles immediately after use

22 Source: Norm DWA-A 704 / State 2007, IQC-Card 9, Sheet 1

Monitoring of Thermoreactors
DWA A704: Monitoring of heating
device/thermoblock
Check the required temperatures of 100 C and/or 148 C ( 3 C).
Alternatively, also other/further frequently used temperatures can be tested.
Step 1: A round cell is inserted into a borehole of the thermoblock (any).
The cell is filled half with glycerine (H2O free), oil or sand and is equipped
with a thermometer that is calibrated for the respective temperature range.
Step 2: The thermoblock is connected and is switched on.
Step 3: The temperature control is adjusted to the temperature that is to
be checked and the clock is set to approx. 1 hour.
The temperature is measured at the earliest 10 min after the control lamp
extinguishes.

24 Source: Norm DWA-A 704 / State 2007

Example: Documentation
Heating devices/thermoblock
Wastewater system: Treatment plant Hoppenstdt

1 2 3 4 5 6
Date Thermoblock 100 3C 148 3C _____ 3C Assessment
no. Signature
20 January 2011 1 102 148 ok
Peters
20 January 2011 2 102 148 ok
Peters
20 January 2011 3 100 150 ok
Peters
04 January 2012 1 101 148 ok
Franu
04 January 2012 2 100 140 QT* not achieved,
customer service
Peters
04 January 2012 3 95 138 QT* not achieved,
customer service
Peters
*QT: Quality target
25 Source: Norm DWA-A 704 / State 2007, IQC-Card 9, Sheet 2
Thermoreactors
Thermoreactor TR 320
(without pressure and temperature
measuring function)
12 X 16 mm
Ord. no. 1.71200.0001

Thermoreactor TR 420
24 X 16 mm
Ord. no. 1.71201.0001

Thermoreactor TR 620
2 separate blocks with each 12 X 16 mm
Ord. No. 1.71202.0001

26
Thermosensor for Spectroquant
Thermoreactor
The thermosensor measures the current temperature in the borehole of
the thermoreactors und compares this temperature with the specified
temperature
For documentation issues, these test results can be transfered to a PC.

27
Documentation

The header reflects type and firmware

version: TR 620, V1.02

Line 2 shows the set reference temperature

(Tref = 100.0 C) whereas the following lines
represent the measured points to check

In this example, the temperature was

recorded after reaching the constant
temperature every minute.

28
Monitoring of photometers
Monitoring of photometers
What is included?
1. Checking the hardware
- Who is in charge? Documentation
- Define test intervals, tolerances
2. Checking the software
- current update of methods
- Who is in charge? Documentation

30
Monitoring of photometers
What is suggested in the Norm (DWA A 704)?
DWA A 704: The device must be checked annually according
to the instructions manual."

Optical system check ok?

no yes

Troubleshooting using the manual Documentation of the monitoring

and e.g. PhotoCheck

31 Source: Norm DWA-A 704 / State 2007

Example: Documentation of Monitoring

Treatment plant Hoppenstdt

1 2 3 4
Testing equipment Assessment Measure Date
signature
Photometer Maintenance by customer Protocol see maintenance 16 February 2004
service; document Franu
ok
pH-electrode Documented in equipment log See there 04 January 2004
Peters

Oxygen measuring device Electrode defective Electrode replaced 16 February 2004

Peters

32 Source: Norm DWA-A 704 / State 2007, IQC-Card 9, Sheet 1

Monitoring of photometers

DWA A 704 Merck Millipore

The photometers must be checked offers a test solution kit for a periodic
annually in their functions according to hardware check (e.g. each 6-8 weeks)
the instructions manual." cheap and fast operation

= Minimum requirement = more reliability through

periodic control

33
 Analytical Quality Assurance with Merck- AQA 1
Monitoring of Photometers (Hardware Check)

Spectroquant PhotoCheck, Highly stable color solutions (Expiry: 3 Years).

1.14693.0001 Absorption tests at different wavelengths with variable
concentrations
Batch-specific certificates with target values and
tolerances
A check in a reference spectrophotometer which
itself is monitored using primary standards (NIST
Standards) confirms that this checking medium can be
traced back to international standard
Documentation of test results via transfer to printers
and PCs
NOVA and Pharo Photometer display the AQA state

34
Example: Lot Certificate PhotoCheck
Art.Nr. / Cat.No. / Art. Nro. 1.14693.0001
Charge-Nr. / Lot no. / Lote nro. HC126311
Verwendbarkeit / Expiry date / Fecha de cad. 31.07.2014
Photometer / Photometer / Fotmetro Referenz / Reference / Referencia
Prfer / Tester / Verificador Fr. Brandner
Datum / Date / Fecha 29.09.2011
Datei / File / Fichero 114693001_HC126311_EN

Prflsung fr / Check Solution for / Solucin de control para: 445 / 446 nm

Lsung / Ext. 10-mm Messwert Extinkion Rund- Toleranz / Minimum / Mnimo Maximum / Mximo
Solution / Kvette / kvette / Result absorbance* Tolerance /
Solucin Abs. 10-mm round cell / Reultado Tolerancia
cell / absorbancia cubeta redonda
Abs. cubeta
de 10mm
445/1 0,149 0,201 0.020 0,181 0,221
445/2 0,369 0,498 0,030 0,468 0,528
445/3 0,742 1,002 0,040 0,962 1,042
445/4 1,114 1,504 0,050 1,454 1,554
* This value has been calculated from the absorbance of the 1 cm cell using the path length of the round cell and is
entered as the desired.

35
Documentation

36
Analytical Quality Assurance with
Spectroquant
Users are assisted in GLP-compliant operations by Spectroquant NOVA and
Pharo Photometer

Instrument supported Analytical Quality Assurance

Photometer SelfCheck
AQA1: Checking the photometer
AQA2: Checking the complete system
AQA3: Checking the influence of the sample [matrix check]

37
Documentation and reminder of AQA1

38
CertiPUR
UV/VIS standards
Reagents for Monitoring according to international standards
Pharmacopeia
USP, JP
DIN EN ISO
ASTM

39
Zero adjustment
Zero adjustment What is to consider?

Master cell
Deionized water
Zero adjustment will be saved by the photometer (NOVA, Pharo)
We recommend to check zero adjustment once per week and if necessary
to perform it again.

41
Saving zero adjustment with date

42
When to repeat a zero adjustment?

We recommend to repeat the zero adjustment in the following cases:

If the photometer was subject to mechanical stress such as strong shock
or transport
If the ambient temperature changed by more than 5 C since the last zero
adjustment
After the lamp was replaced
At least once per week
If a new cell type (different manufacturer, different glass type is used)
Basically each time you want to measure with the highest possible
accuracy

43
Internal Quality Control (IQC)

Internal Quality Control ensures that the handling, the reagents used
and the measuring equipment are checked regularly as follow:

Check of the reproducibility of a sample by multiple determination

Measuring of standard solutions to check the procedure, the measurement
equipment and reagents
Check of plausibility by dilution and standard addition of the sample
Equivalency measurements by means of operating methods from other
wastewater systems (e.g. within the scope of wastewater treatment plant
neighbourhoods) and/or participation in interlaboratory tests
Parallel analyses as a comparison to reference methods
Control and maintenance of the testing medium (adjustment, calibration)
including the sampling devices according to the manufacturers information
Control of the reagents storage life
44 WFA Professional Quality Assurance | 2012 Source: Norm DWA -A 704 / State 2007
Thank You