ICH M10 Bioanalytical Method Validation: Requirements, Acceptance Criteria and What Changed from FDA 2018

Scientifically reviewed by: Dr. Abdulnaser Alsharaa, PhD Bioanalytical Chemistry, Bioanalysis Department Head, PharmaPrimes

Short answer: ICH M10 is the harmonised guideline for bioanalytical method validation and study sample analysis. It covers methods that measure drugs and their metabolites in biological matrices in GLP toxicokinetic studies, nonclinical PK studies used as surrogates for clinical studies, and all phases of clinical trials, including bioavailability and bioequivalence (BA/BE) studies. It sets separate requirements for chromatographic methods, such as LC-MS/MS, and for ligand binding assays (LBAs). Biomarker assays and immunogenicity methods are outside its scope. (ICH M10 as issued by FDA, section 1.3)

Most bioanalysts know the headline numbers. Problems come from applying chromatographic criteria to an LBA, missing an element that M10 added, or relying on the FDA 2018 guidance as if nothing had changed. This guide puts the criteria side by side and identifies the practical changes.

ICH M10 status and implementation dates

For studies under way when M10 took effect, EMA explains when validations performed under its previous guideline may remain acceptable. (EMA)

Where the FDA 2018 guidance stands

FDA’s guidance on bioanalytical method validation for biomarkers states that the May 2018 bioanalytical method validation guidance will be withdrawn when the biomarker guidance is published. (FDA, BMV for Biomarkers) The biomarker guidance was issued in final form in January 2025. (IQVIA Laboratories) The 2018 guidance page on fda.gov still shows it as “Final”, with content dated 2020. (FDA) For drug and metabolite assays, use M10 as the reference.

What ICH M10 covers, and what it does not

• Covered: quantitative chromatographic methods (LC or GC, typically with mass spectrometry) and LBAs used for regulatory decisions on safety, efficacy or labelling.
• Not covered: biomarkers and immunogenicity methods. For exploratory studies not used for regulatory decisions, the applicant decides the level of qualification. (M10, 1.3)
• ICH M10 vs ICH Q2: M10 applies to measuring drugs in biological samples such as plasma. ICH Q2 applies to analytical procedures for drug substances and products, such as assay and impurity testing.

Full validation elements

For chromatographic methods, a full validation includes:

• selectivity and specificity;
• matrix effect;
• calibration curve and range (LLOQ to ULOQ);
• accuracy and precision;
• carryover;
• dilution integrity;
• stability;
• reinjection reproducibility.

For LBAs, the elements are:

• specificity and selectivity;
• calibration curve and range;
• accuracy and precision;
• carryover;
• dilution linearity;
• stability;
• parallelism, when suitable study samples are available.

The validation matrix should match the study matrix, including anticoagulant and additives. (M10, 2.2.1)

Acceptance criteria: chromatographic methods vs ligand binding assays

All values are from the M10 text (sections 3, 4 and 5). Criteria do not transfer between the two columns.

Study sample analysis: what reviewers check

• Run composition (chromatography): blank, zero, at least 6 calibration levels, and low, medium and high QCs in duplicate or at least 5% of the number of study samples, whichever is higher. Study samples are always bracketed by QCs. (M10, 3.3.1)
• Dilution QCs: runs with diluted samples include dilution QCs. (M10, 3.3.2)
• Overall QC performance: overall accuracy and precision across accepted runs are reported. In comparative BA/BE studies, failure may lead to rejection of the data. (M10, 3.3.2)
• No pharmacokinetic reanalysis in BE: samples are not reanalysed because a concentration does not fit the expected profile. (M10, 3.3.4)
• ISR coverage: ISR is expected in all pivotal comparative BA/BE studies, the first clinical trial, pivotal early patient trials, first or pivotal hepatic or renal impairment trials, and nonclinical studies within scope. The minimum is 10% of the first 1,000 samples plus 5% of samples beyond 1,000. (M10, section 5)
• Cross validation: required when data come from different methods or laboratories within a study, or are combined across studies. Bias is assessed with QCs and study samples (n of at least 30), for example by Bland-Altman or Deming regression. (M10, 6.2)

What changed from the FDA 2018 guidance

Sources disagree on how much changed. One industry summary says the method validation and study sample acceptance criteria did not change (ProPharma Group), while others list practical differences (Bioanalysis Zone; Frontage Laboratories). The core numbers are familiar. The differences are in what must be done and documented:

Checklist before validation starts

• Confirm the assay type, then apply only that section’s criteria.
• Match the validation matrix to the study matrix, including anticoagulant.
• For LC-MS/MS, use a stable isotope-labelled internal standard whenever possible. (M10, 3.1)
• Plan stability so that validated storage periods and freeze-thaw cycles cover the study.
• For fixed-dose combinations, run matrix stability with all dosed compounds present. (M10, 3.2.8)
• Define reanalysis, reinjection and reintegration rules in an SOP or protocol before analysis begins.

How PharmaPrimes supports bioanalytical method validation

PharmaPrimes validates bioanalytical methods according to applicable ICH M10 requirements, on platforms including LC-MS/MS, ELISA and qPCR systems with Watson LIMS, for small molecules, peptides including GLP-1 analogues, monoclonal antibodies and other biologics. Explore our bioanalytical method validation services, integrated with clinical conduct for BE/BA studies.

Frequently asked questions

What does ICH M10 stand for?

M10 is the ICH guideline titled “Bioanalytical Method Validation and Study Sample Analysis”. The “M” denotes ICH’s multidisciplinary guideline series.

What is the latest version of the ICH M10 guideline?

The Step 4 guideline of 24 May 2022, supported by ICH Q&As and January 2024 training material. Check the ICH website for any later update.

What are the FDA guidelines for bioanalytical method validation?

FDA adopted ICH M10 as guidance in November 2022 for drug and metabolite assays. Biomarker assays are covered by a separate FDA guidance issued in 2025.

What are the acceptance criteria under ICH M10?

For chromatographic methods: accuracy within ±15% (±20% at LLOQ) and precision no more than 15% CV (20% at LLOQ). For LBAs: ±20% and 20% CV, widening to ±25% and 25% at LLOQ and ULOQ, with total error no more than 30% (40% at the limits).

Does ICH M10 apply to biomarkers?

No. Biomarker assays and immunogenicity methods are outside its scope.

Discuss your validation

Planning a BE study, a method transfer or a partial validation under ICH M10? Discuss your bioanalytical method validation with our bioanalysis team.

References

1. ICH. M10 Step 4, 24 May 2022. https://database.ich.org/sites/default/files/M10_Guideline_Step4_2022_0524.pdf
2. FDA. M10 guidance, Nov 2022 (full text). https://www.hhs.gov/guidance/sites/default/files/hhs-guidance-documents/FDA/50093493fnl_M10-Bioanalytical-Method-Validation-and-Study-Sample-Analysis_Jan-2025-update.pdf
3. EMA. Implementation strategy of ICH M10. https://www.ema.europa.eu/en/documents/scientific-guideline/implementation-strategy-ich-guideline-m10-bioanalytical-method-validation_en.pdf
4. ICH. M10 training material, Jan 2024. https://database.ich.org/sites/default/files/ICH_M10_EWG_Training_Material_2024_0127.pdf
5. FDA. Bioanalytical Method Validation for Biomarkers. https://www.fda.gov/media/185276/download
6. FDA. Bioanalytical Method Validation, May 2018. https://www.fda.gov/files/drugs/published/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf