Differing bioanalytical method validation and sample analysis requirements among global

health authorities have challenged the drug development process, causing inefficiencies
and uncertainty.

In 2010, regional scientific associations began collaborating in earnest to discuss the

challenges created by country-to-country differences. These inter-association discussions
resulted in several publications that identified the need for a global bioanalytical guideline
[1,2] and the creation of the Global Bioanalysis Consortium (GBC). Subject matter experts,
including scientists from Labcorp Drug Development (NJ, USA), participated in the GBC to
develop and publish harmonized best scientific practices.

From Bioanalytical Method Validation Guidance to ICH M10

In subsequent years, additional countries published their version or updated versions of
their Bioanalytical Method Validation Guidance/Guidelines – adding to the complexity and
sometimes conflicting expectations that bioanalysts had to fulfill to achieve compliance. In
2016, Japan’s Ministry of Health, Labour and Welfare (MHLW) and Pharmaceuticals and
Medical Devices Agency (PMDA) submitted a request to the International Council for
Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH)
Management Committee for a harmonized bioanalytical guidance. The ICH Management
Committee accepted and published the M10 process concept paper in October 2016, and
the Expert Working Group comprising ICH member country regulators, industry members
and observers first met a month later.

Over the next three years, the Working Group met and considered the requirements for a
global harmonized guideline; publishing a draft for public consultation in 2019. Labcorp
Drug Development scientists participated in a number of regional organization activities,
including the Chinese Bioanalysis Forum, European Bioanalysis Forum and American
Association of Pharmaceutical Scientists, which developed and submitted comments to the
Working Group. Unable to meet face-to-face during the COVID-19 restrictions, the Expert
Working Group continued to meet virtually to review the public’s comments and
recommendations on the draft Guideline. On May 24, 2022, the Expert Working Group was
able to meet in Athens, Greece, where the regulatory members signed the final version of
the guideline. ICH published that guideline, “Bioanalytical Method Validation and Study
Sample Analysis”, on June 8, 2022.

Labcorp Drug Development has closely monitored the progress of the ICH M10 Expert
Working Group and developed implementation plans to ensure compliance through the
transition from regional guidelines to ICH M10. The Expert Working Group released a
“Frequently Asked Questions” guide and is expected to release other training material over
the next few months, and Labcorp Drug Development will continue to refine our plans based
on any clarifying information in these new materials.

As a global provider of bioanalytical services, Labcorp’s globally harmonized standard

operating procedures have been developed over a number of years to accommodate
various regional regulations. Here, Labcorp Bioanalytical Services subject matter experts
representing China, the European Union and the United States of America offer their
perspectives on the impact of changing from current regional expectations to ICH M10.
 ICH M10 in China
The National Medical Products Administration (NMPA; formerly known as the China Food
and Drug Administration) has been engaging in significant regulatory reform over the past
10+ years. NMPA issued a series of guidance for bioanalysis to bring its regulations in line
with international standards. In December 2011, China issued a guidance for clinical trial
bioanalytical laboratory management. In 2015, the China Pharmacopeia issued a guidance
for bioanalytical method validation, similar to U.S Food and Drug Administration (FDA) and
EMA guidance, and it was updated in 2020 to reflect the latest regulatory requirements [3].

China joined ICH as a new Regulatory Member in May 2017. NMPA has implemented
reforms to align China’s regulations with global standards adopting 361 guidance
documents from ICH, FDA, EMA and World Health Organization (WHO). Chinese
representatives joined the Expert Working Group (EWG) for the development of the ICH
M10 Bioanalytical Method Validation Guideline. The NMPA Center of Drug Evaluation
(CDE) ICH Working Group invited public consultation for the draft ICH M10 guideline in April
2019, receiving more than 100 comments, which were mostly from the China Bioanalysis
Forum (CBF) and pharmaceutical industry companies in China.

There are two main types of clinical studies that require regulated bioanalysis support
locally in China. First, to register a new drug in China, global pharma must conduct both a
pharmacokinetic (PK) study and a Phase three study in a Chinese population. Global
submissions require bioanalysis executed in China to be consistent with that executed in
other regions. The new ICH M10 guideline is expected to harmonize expectations and
streamline global drug development.

Second, when there is a Global Multi-Regional Clinical Trial (MRCT) with patients in China,
Chinese bioanalysis is an essential part of the global trial. ICH M10 cross validation
requirements to combine the data generated from different laboratories overcome
differences in the current regional requirements with its harmonized approach:

 ICH M10 indicates cross validation is required to combine related data when multiple
bioanalytical methods and/or bioanalytical laboratories are involved.
 ICH M10 suggests that if possible, cross validation should be assessed by measuring the
same set of QCs at least in triplicate and study samples (if available) that span the study
sample concentration range with both methods, or in both laboratories.
 ICH M10 recommends that bias can be assessed by Bland-Altman plots or Deming
regression. Other methods appropriate for assessing agreement between two methods
(e.g., concordance correlation coefficient) may be used as well. Alternatively, the
concentration vs. time curves for study samples could be plotted for samples analyzed by
each method to assess bias.

As a global service provider, we are looking forward to the full implementation of ICH M10
to streamline global drug development programs.

ICH M10 in the European Union

The current EMA guideline on bioanalytical method validation [4] became effective in
February 2012, was the first to publish separate expectations for ligand binding assays
(LBA); recognizing that they differ substantially from chromatographic analytical methods.

The scope of the EMA guideline is wide, providing ‘recommendations for the validation of
bioanalytical methods applied to measure drug concentrations in biological matrices
obtained in animal toxicokinetic studies and all phases of clinical trials’. Within ICH M10, the
scope is more specifically defined to ‘nonclinical toxicokinetic (TK) studies conducted
according to the principles of GLP, nonclinical PK studies conducted as surrogates for
clinical studies, and all phases of clinical trials, including comparative
bioavailability/bioequivalence (BA/BE) studies, in regulatory submissions’. ICH M10 also
clarifies that full validation is expected for the primary matrix supporting the regulatory
submission(s) and regulatory decisions regarding safety, efficacy or labelling; for evaluation
of drug (and/or metabolites) levels in additional matrices, ICH M10 asks that assays are
‘validated as necessary’. This allows scientific freedom to validate according to the intended
purpose of the data.

An area where there is a notable difference between the EMA guideline and ICH M10 is
method development. The EMA guideline does not contain a dedicated section on method
development, but does discuss assessments that would typically be performed during
method development, such as (surrogate) matrix selection in cases where there are related
endogenous compounds, and establishment of a minimum required dilution (MRD) for LBA.
ICH M10 contains a dedicated section for method development, though it is limited to
general considerations for establishing that a bioanalytical method is suitable for its
intended purpose and ready for validation, and includes a statement that method
development ‘does not require extensive record keeping or notation’.

There is an important difference between the EMA guideline and ICH M10 in the sections
on cross-validation. Both EMA and M10 asks that the same set of quality control (QC)
samples and/or study samples be analyzed with the different bioanalytical methods and/or
by the different bioanalytical laboratories. However, EMA asks for a direct comparison of
cross-validation results with a pass/fail approach, whereas ICH M10 does not contain any
specific acceptance criteria for cross-validation and rather asks for analysis of any biases
between methods or laboratories, which will allow more latitude to combine data from
across platforms and/or laboratories providing that any identified biases are properly
addressed.

For the majority of method validation assessments, such as selectivity, accuracy and
precision, dilutional linearity, stability, there is little conflict between what is asked for in the
EMA guideline and in ICH M10, the majority of differences being additional requirements in
M10. In assessing selectivity for ligand binding assays, EMA asks for evaluation in 10
sources of sample matrix at or near the lower limit of quantitation (LLOQ), whereas ICH
M10 recommends assessment at both LLOQ and high QC levels. For evaluation of
dilutional linearity, ICH M10 asks for at least 3 independently prepared dilution series,
whereas the number of preparations is not specified in the EMA guideline.

Perhaps the most impactful differences are found in expectations for assessment of analyte
stability. ICH M10 states that for each concentration tested for stability, the bulk sample
should be divided into a minimum of 3 aliquots that will be stored, stressed and analyzed.
There is no such requirement to stress independent aliquots in the EMA guideline. The
EMA guideline considers evaluation of analyte stability using the low and high QC levels of
the assay sufficient, whereas ICH M10 will require evaluation of a higher concentration level
stability QC sample(s) in cases where study sample concentrations are consistently higher
than the ULOQ of the calibration curve.

There are also some welcome clarifications. For preparation of QC samples, EMA asks for
at least 5 QC levels in LBA (anticipated LLOQ, less than 3 times the LLOQ, mid, high and
anticipated upper limit of quantitation (ULOQ)), which has led to some ambiguity in
expectations in the placement of the medium QC: some European agencies have accepted
placement at the geometric mean, whilst others have insisted on placement around 30-50%
of the calibration curve range, as stated in the chromatography section; ICH M10 brings
harmonization in placement of the medium QC around the geometric mean of the
calibration curve range.

ICH M10 in the USA

Two additions stand out in ICH M10 related to general principles that are not seen in the
FDA 2018 guidance [5] and address animal use and clinical trial subject safety. With regard
to animal use, M10 emphasizes developing ‘drugs in accordance with the principles of 3Rs
– Reduce, Refine, Replace – for animal studies’. For clinical trial subjects, Good Clinical
Practices have historically emphasized the safety of trial subjects and been applicable as
part of bioanalysis. However, M10 takes the additional step of emphasizing that the
reanalysis of “specific study samples for the purpose of a safety investigation” is acceptable.

A few notable changes from the FDA bioanalytical method validation (BMV) to the ICH M10
impact both ligand binding and chromatographic assay practices. For example, no QC
rejection criteria for accuracy and precision (A&P) runs were established in the FDA BMV to
allow the within and between run calculations to use the full data set. ICH M10 reverts to
including acceptance criteria to QCs in A&P runs but requires all QCs to be included in
within and between run calculations with the exceptions of QCs with documented analytical
errors. Additionally, M10 also includes more extensive instruction for evaluation of lipemic
and hemolyzed matrices, matrix effects and includes testing in relevant patient populations.

When considering therapies that are endogenous analytes, ICH M10 is much more
prescriptive with many more details and sections than in the FDA BMV, including four
sections on addressing calibration curves when there is no true blank matrix, approaches to
preparing QCs, experimental design for selectivity, recovery and matrix effects, parallelism,
stability, and accuracy and precision.

Additionally, ICH M10 addresses and clarifies multiple topics of current debate. The ICH
M10 guideline states that stability determinations for bench top and/or autosampler stability
are not to be conducted using an additive approach, but rather a single long duration test
should be performed and reported. Stability at one facility can be used as representative of
stability at another facility provided there is adequate data and documentation.
Chromatographic assays stock solutions made prior to the expiration of the reference
standard are considered valid and the stock solution stability is that established for the
solution.
There are few differences around sample analysis practices for ICH M10 vs the FDA BMV.
While conceptually similar, ICH M10 specifies QCs bracket study samples, whereas the
FDA BMV described that QCs need only be interspersed with study samples. When
considering incurred sample reanalysis (ISR), in the FDA BMV for clinical studies ISR was
applicable to bioequivalence (BE) and pivotal pharmacokinetic or pharmacodynamic
studies, whereas in ICH M10, the scope of ISR is expanded to BE, bioavailability (BA), first
clinical trial, pivotal early patient trials and trials with patients with impaired hepatic and
renal function. In both cases, the term pivotal is used and in the absence of clear definition
may result in unneeded effort as sponsors take risk-adverse approaches.

Whilst FDA had already moved the needle to include additional reporting in 2018, ICH M10
went even further and includes a number of additional reporting content components. Many
additional requirements for documentation and reporting are related to bioavailability (BA)
and bioequivalence (BE) studies. Many additional details appear to be related to decreasing
the need for on-site inspections and increased ease in detecting fraud for these pivotal
studies. In addition to the report requirements, ICH M10 requires when BA or BE studies
are in a filing that the sponsor include within Section 2.6.4/2.7.1 of the Common Technical
Document details on regulatory inspections of the bioanalytical site in the 3 years prior to
the study sample analysis and one-year post analysis.

Adhering to changing requirements

In conclusion, ICH M10 integrates the practices of health authority bioanalytical regulations
from around the world into a single standard, making it much easier for bioanalytical work to
be performed and meet global regulatory expectations. As this article was nearing
completion, EMA published its adoption of ICH M10 with an effective date of January 21,
2023, establishing the first of many regional implementation dates. Labcorp Drug
Development staff have been active in the discussions of harmonized global regulations for
more than a decade and are prepared to deliver bioanalytical services that meet the new
requirements of ICH M10.