Workshop on laboratory basics and fundamentals of ISO Quality Management Standards March 21-22, 2018, Kyiv, Ukraine Practical work in groups

1. EUROPEAN UNION WATER INITIATIVE PLUS
FOR THE EASTERN PARTNERSHIP
Support of Water Policy Reforms towards Integrated Water Resources Management
Workshop on laboratory basics and
fundamentals of ISO Certification

2. Day 2 Thursday 22/03/2018
9:00 Wrap up Day 1
9:15 Practical work in 3 groups
incl. breaks
12:00 Lunch break
13:00 Discussion of outcomes, Q&A
14:00 Occupational safety aspects
15:00 Coffee break
15:20 IT requirements
16:00 Summary, Feedback, Award of Certificates
16:30 Closure of the Workshop
2

3. Wrap-Up
 What were your most important impressions
yesterday?
 What was absolutely new?
 What was a boring repetition?
3

4. Group composition
 18 experts from laboratories
 Prepared numbers in hat
Please pick one item which indicated your group nr.
4

5. Introduction to exercise
 Set-up of a new method in the laboratory
 Walk through all stages from decision to
establish/amend the method to first analyses
 Trainers will provide you a frame for the group work
and give additional information
 Reflection in plenum
5

6. PDCA Cycle

7. Introduction to exercise
 Assumption for today
Your laboratory decides to establish a
spectrophotometric method to determine
Ammonium NH4
+ in water samples
7

8.  What is needed to start planning?
 2 minutes for cogitation
 Call your ideas
Exercise 1
8

9. Quality component Parameter Surveillance
monitoring
First survey
Surveillance
monitoring
Repetition
Operative
monitoring, time
frame
Survey frequency
[…] pollutants Ammonium 1 year 5 years 1 year Once per month
Exercise 2
Parameter CAS Nr. Standard method Minimum Limit of
Qualification LOQ
[…]
Ammonium NH4-N - ISO 7150-1 Dec. 87 0,01 mg N/l
[…]
Excerpt from deliberate National Implementation of 2000/60/EC – national ordinance 465/2010:
Annex 2, table 2.1.1.
The ordinance defines 100 ground water sites in the country, to be listed in Annex 3 (not enclosed)
Annex 12, Table 2.1.3:

10.  Headlines
• Scope and field of application
 Substance, type of sample, range, LOD, sensitivity
• Principle
• Reagents
 Definition of substances, procedures to come to working solutions,
calibration solutions etc.
• Apparatus
• Sampling and samples
• Procedures
 Test portions, preparation of test solutions, determination
• Calibration
• Results calculation
• Interferences
ISO 7150/1

11.  Establish the objectives and processes necessary to
deliver results in accordance with the expected
output/ target, goals (PDCA cycle)
 Check technical details (norm, scope, instrumental needs)
 What can be anticipated for the next steps?
• Who is going to do the work?
• In which timeframe?
• With which budget?
Elaborating a plan
11

12. We DO!
12
 Implement the plan, execute the process, make the
product. Collect data for charting and analysis in the
following „check and „act steps
• Retrospection to yesterday s presentation …

13. 1st calibrations
13
R² = 0.9995
0
0.2
0.4
0.6
0.8
1
1.2
0 100 200 300 400 500 600

14. Calibration range
 Calibration range 1 (1-1000 µg/L NH4-N)
• LOD: 16 µg/L
• LOQ: 55 µg/L
 Calibration range 2 (1-100 µg/L)
• LOD: 2,3 µg/L
• LOQ: 7,5 µg/L
14

15. Calibration
15
R² = 0.9996
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0 5 10 15 20 25 30

16. Calibration
Absorbance
Blank 1 (pure water) 0,038
Blank 2 (pure water) 0,040
Blank 3 (pure water) 0,039
Blank 4 (tap water) 0,043
Blank 5 (tap water) 0,044
Control standard, 20 µg NH4-N 0,079
16
A series of blanks have been analysed. A control standard has been prepared individually,
from weighing the raw material and diluting it down to 20 µg.

17. Calibration
17
R² = 1
0
0.02
0.04
0.06
0.08
0.1
0.12
0 5 10 15 20 25 30

18. Statistics
 Calibration function
• Correlation: 0,99991
• Sensitivity: 0,004
• Axis intercept: 0,039
• LOD: 0,14 µg/L
• LOQ: 0,69 µg/L
18

19. Control charts
19
0.003
0.0032
0.0034
0.0036
0.0038
0.004
0.0042
0.0044
0.0046
0.0048
0.005
0 5 10 15 20 25 30
Absorbance vs. Nr. of Samples
0.003
0.0032
0.0034
0.0036
0.0038
0.004
0.0042
0.0044
0.0046
0.0048
0.005
0 5 10 15 20 25 30
Absorbance vs. Nr. of Samples
19
19.5
20
20.5
21
21.5
0 5 10 15 20 25 30
Concentration [µg/L] vs. Nr. of Samples

20. Validation
EN ISO/IEC 17025
Validation is the confirmation by examination and the
provision of objective evidence that the particular
requirements for a specific intended use are fulfilled.
20

21. Validation
volumetric instruments
balance room conditions
traceable reference material and standards
expiry dates of standards
frequencies of routine checks defined
documentation

22. Balances
proper equipped balance room
• negligible vibrancies
• temperature control
 intermediate checks with calibrated weights
• UBA Austria: frequency of checks: once per month (each
balance)
 daily internal calibration acc. to manufacturers instructions
 logging forms balance book
Validation

23. Validation
 material or substance of which
properties are sufficiently
homogeneous and well
established
 to be used
• for the calibration of an
apparatus
• for the assessment of a
measurement method
• or for assigning values to
material
Certified Reference Material (crm):
 reference material
accompanied by a certificate
 certified by a procedure which
establishes traceability to an
accurate unit in which the
property values are expressed
 each certified value is
accompanied by an uncertainty
at a stated level of confidence
Reference Material (rm):

24. laboratories need to ensure that
• valid methods of measurement are selected
• and that there is clear evidence that the methods
are capable of measuring what needs to be
measured
Validation

25.  description of requirements
 determination of meaningful method statistics (detection
limits, quantification limits, working range, recovery rates,
…
 comparison of requirements with method derived statistic
data
Is method capable to fulfil requirements?
YES/NO
Standardized methods need not to be validated
– but method characteristics for the application have to be determined
Validation

26.  participation in laboratory intercomparisons
 running of trial tests with samples of known values
 (certified) reference materials or home-spiked control
samples
• determine uncertainties of measurement
• limits of detection
• confidence limits
 method comparison against another technique
 spiking of real samples → recovery rates
Validation

27. Validation
statistic data
Accuracy
Precision
• Repeatability
• Intermediate precision
• Reproducibility
Selectivity
information about
Systematic errors
(Do test result and true value
agree?)
random errors
short-term variability (within laboratory)
long-term variability (within laboratory)
precision between laboratories
accurate measurement in presence
of interferences

28. Validation
statistic data
Recovery rate
Detection limit
Quantification
limit
information about
Yield
LOD = 3s
Lowest concentration of
analyte detected with
acceptable level of
confidence (e.g. LOQ = 10s)

29. Validation
Statistic data Information about
linearity relationship analyte response
to concentration
robustness influence of small variations in
procedure?
working range acceptable uncertainty within
range
sensitivity response

30. 30

31. EU MEMBER STATES AND EUROPEAN COMMISSION:
EASTERN PARTNERSHIP (EaP) COUNTRIES:
BELARUS MOLDOVA UKRAINEARMENIA AZERBAIJAN GEORGIA
AUSTRIA FRANCE
INTERNATIONAL ORGANISATIONS: