BIOANALYTICAL METHOD VALIDATION AND STUDYSAMPLE ANALYSIS
M10

Frequently Asked Questions (FAQs)

Document date: 25 May 2022

To support the implementation of ICH M10, the Expert Working Group has developed a seriesof FAQs.

Guideline
SectionQuestions

Answers

1

The guideline states that it isapplicableto“nonclinicalpharmacokinetic (PK) studies
conducted as surrogates for clinicalstudies…” Please provide an
example for such studies.

One example which includes nonclinicalPK data to support human dosing is
rescue agents for acute radiation
syndromes or anthrax etc., under theAnimal Rule (FDA, United States).

2In situations where a matrix is Yes, as long as the use of the surrogate
unavailable (e.g., shortage, 3Rs - matrix meets the requirements of theReduce, Refine, Replace) can a guideline, including accuracy andsimilar surrogate matrix (e.g., precision, lack of interferences, etc. andhuman plasma) be used to dilute the dilution quality control samples

2, 3, 4

samples?

(QCs) are processed in the same way.
The rationale needs to be well justified
because the approach might bequestioned.

When adding a new QC
concentration level during study
sample analysis without changing
the calibration curve range in either

The precision and accuracy of the new
QC concentration level should bedemonstrated before use in study sample
analysis. This can be documented either

chromatographic assays or ligand as a partial validation or as a note to thebinding assays, is it necessary to bioanalytical report.
validate the new QC concentrationlevel with a partial validation?
3Is it acceptable to demonstrate the Yes, this is applicable for both method
absence of analytical interference validation and study sample analysis.
of the IS itself, any impurities or its
isotopic stability based on theanalytical results of the zerosample?

3

For long-term stability, does afailed time-point mean you should
not continue with longer timepoints?

3

Can the physicochemical properties Yes, but if co-elution of the relatedof the related substances be used to substance and the analyte is not
2

Additional time-points can be evaluated.
Any failure should be investigated toidentify the root cause and the impact on
the stability assessment.

justify that the related substances excluded, additional investigations are
do not co-elute or interfere with the needed to demonstrate chromatographic
analyte measurement during mass separation (e.g., for isomers). If thespectrometry (MS) analysis?
analyte and the related substance coelute,matrixeffect(ionsuppression/enhancement) and backconversion should be evaluated.
3

What does the guideline mean with The diluted concentrations should fallrespect to “concentrations” in the within the validated calibration curve
following sentence: “The dilution range.
factor(s) and concentrations applied
during study sample analysis
should be within the range of thedilution factors and concentrations
evaluated during validation”?

3

What is the purpose of measuring To confirm the QCs were correctly
the concentration of the QC at time prepared. Stability in the matrix (e.g.,zero?
bench-top, long-term, freeze-thaw)
should be evaluated by comparing with
the nominal value.

3

How is the accurate preparation of By comparing two independently
the stock solution verified?
preparedstocksolutions
anddemonstrating that the difference of theirmeasured responses is within 5%.
% difference=(Stock solution 1 - Stocksolution 2)/(mean value) × 100

4Is there a requirement to test
specificity in validation with anirrelevant
immunoglobulin
molecule when the analyte is an
immunoglobulin and the assay
contains analyte specific reagents

There is no requirement to assess
specificity in validation with anirrelevant immunoglobulin as long as thespecificity of the reagent(s) has been
evaluated
duringreagent
characterisation.

(e.g.,
useof
anti-idiotypic
antibody(ies) as capture and/ordetection reagents)?
5

How should trends of concern or The investigation should be driven by anincurred sample reanalysis (ISR) SOP and should take into account thefailure be investigated?
3

entireprocess,

includingsamplehandling, processing and analysis.
This should also include a scientificassessment of whether there are issues
impacting the bioanalytical method, suchas interferences and instability.
6

Given that M10 allows partial
validation for matrices within
species or same matrix acrossspecies, is an N-in-1 approach(multiple species or matrices in 1validation)
allowedchromatographic methods
nonclinical studies?

forfor4

Possibly this approach may be used.
However, caution should be taken inusing this approach; the rationale needs
to be well justified, because the approach
might be questioned.