ALTERNATIVE APPROACH:

FOR ESTIMATION OF
MEASUREMENT
UNCERTAINTY IN
ANALYTICAL CHEMISTRY

D. Von Mollendorf

Midvaal Water Company

 Overview
• Background
The use of GUM proved to be very successful when studying or quantifying
individual uncertainty components
But it was found that in our case many of the uncertainty estimates were
found to be unrealistic – in most cases under estimated
E.g.
* Standard uncertainty of 5% < long-term precision
(Standard deviation of a series of results)
* Control limits of QC chart: 12%
* Selectivity results of PT for different matrices/concentrations: 83 - 115%
(Excellent z-scores)
* Intra-assay precision (%RSD): 92 - 106%
Also experience challenges using GUM mainly because
- Large number of uncertainty sources in complex testing procedures are
challenging to quantify separately
- Experienced difficulty in determining a complete mathematical model to
describe method.
 Overview
• Objective
Finding a simplification of the GUM approach
Empirical approach? Since the use of empirical approaches for quantitative
test procedures as an alternative to the modelling approach are increasingly
being recognised.
Empirical approaches are based on whole method-performance
investigations, designed and conducted so as to compromise the effects from
as many relevant uncertainty sources as possible

• Results
Nordtest approach was implemented:
* In most cases uncertainty values was found to be higher than previously
found using the modelling approach.
* Proved to have been a better estimation of the real variation of our
methods, mainly due to maximising the probability of including all
uncertainty contributions
* Higher uncertainties was in good agreement with inter laboratory
comparison results and quality control data.

• Recommendations
Nordtest approach - holistic approach - minimise risk of excluding important
uncertainty sources & easy to use already existing data
 Uncertainty Approaches
Definition of the Measurand

Single laboratory Inter laboratory

Model-based ? One procedure ?

Yes No Yes No

Modeling Single lab Interlaboratoy Proficiency

Component by Validation Validation Testing (PT)
Component Within-lab Reproducibility Between Lab
Evaluation reproducility & and Bias Variability
ISO GUM Bias ISO 5725 ISO Guide 43
NordTest TR 537 ISO TS 21748 ISO 13528
 Single Laboratory Approaches
A) Model based approach (GUM, Bottom Up)
• Involves component-by-component evaluation of
uncertainties

B) Single-lab validation approach

• Calculates a typical uncertainty based on whole method-
performance and not directly linked to any particular sample
• Has been formalised in several guides, of which the Nordtest
Uncertainty Guide is the best known
 Inter laboratory approaches
A) Proficiency Testing Approach
• Gives general uncertainty which can be expected from the same
procedure in different laboratories
• Main use: Estimation of uncertainty for a procedure.

B) Inter laboratory Validation Approach

• Use of PT data for estimating measurement uncertainty still under debate
and authoritative references are few.
• But, if a laboratory has successfully participated in an inter-laboratory PT,
it may utilise the results for estimating the measurement uncertainty.
 Comparison of the approaches
Modelling
• Advanced laboratories
- Extra work usually required
- Deep knowledge/competency required
•Danger to under estimations
•Promotes Thinking - more information
Inter-laboratory Validation
• Uncertainty of results obtained using the same
procedure in different laboratories
• Crude uncertainty estimates
Single-lab Validation
• Routine laboratories
- Less extra work required
- Does not need such a high competency level
Realistic uncertainty estimations - usually does
not lead to under estimations, sometimes to
over estimations
• Not as detailed - Does not learn as much
• Takes into account more general uncertainty
contributions such as within-lab reproducibility
& laboratory bias.
PT Approach
• Actually data from your lab not taken into
account - therefore not recommended
•Very crude uncertainty estimation
• Should be used only as first approximation
 Single Laboratory Validation Approach
Measurement accuracy = precision + trueness
Measurement uncertainty= within-lab reproducibility + uncertainty of bias
Combine:
Standard deviation s & an estimate bias b
Measurement uncertainty is the estimated as a root sum of squares of:
 Single Laboratory Validation Approach
• Provided that measurements are under statistical control,
uncertainty estimates obtained using this approach are
applicable for all measurements within the scope of the
measurement procedure.
• Application range of the uncertainty estimates is determined
by the range covered in validation study & on-going quality
control.

• Therefore these investigations should include:

Appropriate within-scope variations, e.g. different levels of
the measurand and different types of test items.
 The Nordtest Approach
Combination of Uncertainty components:

Reproducibility within the laboratory: Rw

(from control samples & other estimations)

Estimation of the method and laboratory bias: u(bias)

(from CRM, PT or Recovery tests)

Combined Standard Uncertainty (uc):

Estimated as a root sum of squares of the two components
(All expressed as % relative uncertainty)
 Cause and Effect diagrams
GUM Nordtest
Identify potential uncertainty sources Determine the uncertainty
estimates contribute to:
Input quantities Bias
Precision
 Reproducibility within the Laboratory Rw
Two different standard deviations can be obtained:
• srw - within-laboratory repeatability standard deviation
Normally not suitable as uncertainty estimate, since it excludes
major uncertainty contributions
• sRw - within-laboratory reproducibility standard deviation
(long-time repetition).
 Method 1:
Control sample covering whole analytical
process
• If control sample covers the whole analytical process and has a matrix
similar to the samples:
* The reproducibility within the laboratory Rw at that concentration level
can simply be estimated from the analyses of the control sample
* If the control sample analyses covered a wide concentration range it
should be used.
• * Combined uncertainty for Rw is calculated by the root sum of squares of
the uncertainty estimates
 Method 2:
Control samples for different matrices and
concentrations
If a synthetic control solution is used for quality control, and the matrix type of
the control sample is not similar to the natural samples:
• Mean Control Chart:
Long term reproducibility contribution - does not include uncertainties
arising from different matrices & sample preparation procedures.
• Range Control chart:
Estimate repeatability from different matrices & sample preparation
processes.
• Combined uncertainty for Rw is calculated by the root sum of squares of the
repeatability and reproducibility uncertainty estimates

Note: The repeatability component is included

two times!!
 Method 3:
Unstable control samples
• If the laboratory does not have access to stable control
samples (e.g. measurement of dissolved oxygen):
* Estimate uncertainty components from repeatability
obtained from a range control chart
* “long-term“ uncertainty component (from batch to batch)
needs expert judgement
 Laboratory and Method Bias
• Possible bias - Not easy to determine the true bias of a
procedure - it needs a lot of parallel measurements under
carefully controlled conditions.
• In this procedure the best estimate of true value is taken into
account - Include sample preparation!
 Laboratory and Method Bias
Calculate bias by combining two uncertainty components

Component accounts Component accounts for

for the average bias of the average uncertainty
the laboratory results of the reference material

Because bias will in most cases vary with different matrix & concentration ranges
* Ideally use several different reference materials/certified materials & several
PT results
* To prevent that bias is grossly under estimated or grossly over estimated
 RMSbias
• Accounts for average bias of the laboratory:

• Every separate bias value (% difference) is calculated as the

value found in the laboratory minus the reference value:
biasi = Clabi - Crefi
Clabi - from repeated measurements
Crefi - preferably include several different reference materials
or inter laboratory comparisons
 u(Cref)
• Accounts for the average uncertainty of the reference values.

• u(Cref) - uncertainty of the nominal/certified value - from COA

If only one CRM is used - include additional term
• u(Cref) - Carried out by inter laboratory comparisons
Estimated from the standard deviation of the participants results
 Laboratory and Method Bias
Four different possibilities:
A) Repeated analysis of the same sample with a reference
procedure - excellent way
B) Repeated analysis of CRM (low uncertainty & high reliability) -
which is very much like your real sample. Very suitable
C) Repeated inter laboratory comparison measurements
Care should be taken: consensus values cannot be called very
reliable → over estimated bias.
• Reproducibility standard deviation sR will be used.
D) Repeated recovery data/spiking results
 Method 1A - Use of one certified
reference material

Uncertainty of certified value u(Cref): 2.21%

Bias of CRM analysed 12 times: Mean: 11.9

100.(11.9 - 11.5)/11.5 = 3.48%

Standard deviation (sbias): 2.2%

Total standard uncertainty

(Root mean square method):
 Method 1B - Use of several certified
reference materials

Mean uncertainty of certified value (Cref):

Bias of 3 CRM’s: 3,48%, -0,9% and 2,4%

Total standard uncertainty

(Root mean square method):
 Method 2 - Use of PT/Inter laboratory
comparison data

Bias of 6 participations: 2%, 7%, -2%, 3%, 6% and 5%

Total standard uncertainty

(Root means square method)::
 Method 3 - From Recovery Tests

The spike of 0.5 mL was added with a

micropipette
Use GUM-approach to quantify

Uncertainty of concentration of spike: 1.0%

Bias of recoveries (6 different matrices: 95 %, 98 %, 97 %, 96 %, 99 % and 96 %

Total standard uncertainty:

Nordtest Approach: Summary

2 Uncertainty components
 Recommendations
• Use the Modeling approach
If you have - Competence and time
- Data on all important influencing quantities

• Use the Single-lab validation approach

If you have - Quality control data & results of participation in
ILC-s or CRM analysis

Simplification of GUM approach for Analytical

Laboratories recommended!
 Examples and literature
• NORDTEST “Handbook for calculation of measurement
uncertainty in environmental laboratories“
• EUROLAB technical report contains:
* Detailed examples, relevant standards, guidelines,
books, internet websites
* Handbook is available from
http://www.nordicinnovation.net/nordtest.cfm as technical
report No. 537

Thank you!