The Guideline That Harmonised Global Bioanalysis
Before ICH M10, bioanalytical method validation was governed by separate guidance documents from the FDA and EMA that, while broadly aligned, differed in specific requirements for acceptance criteria, validation parameters, and reporting expectations. A study submitted to both agencies might require two slightly different validation packages, creating additional work and potential for inconsistencies. The ICH M10 guideline on bioanalytical method validation and study sample analysis, finalised in 2022, replaced both sets of guidance with a single harmonised global standard that applies to all major regulatory markets.
ICH M10 covers the validation requirements for chromatographic assays (LC-MS/MS, LC-UV, and related methods) and ligand-binding assays (LBAs including ELISA and ECL platforms) used in regulated bioanalysis for pharmacokinetic, toxicokinetic, and biomarker studies. Its adoption has significant practical implications for bioanalytical CROs and for the sponsors who commission regulated studies.
Key Changes and Clarifications Introduced by ICH M10
| Topic | Pre-ICH M10 Position | ICH M10 Position |
| Incurred sample reanalysis (ISR) | Required by FDA and EMA guidance; acceptance criteria broadly similar | Harmonised: 2/3 of reanalysed samples must agree within 20% (chromatographic) or 30% (LBA) of original result |
| Parallelism for LBAs | Required by EMA; FDA less prescriptive | Required for all regulated LBAs; failure may indicate matrix effects or non-specific binding |
| Selectivity testing | Test in minimum 6 individual lots of matrix | Minimum 6 lots confirmed; haemolysed and lipaemic matrix testing explicitly addressed |
| Carryover | Addressed in FDA guidance; less explicit in EMA | Explicit requirement: must demonstrate carryover does not affect accuracy; defined acceptance criteria |
| Stability of calibration standards | Covered in guidance; variation in specifics | Detailed stability requirements for stock solutions, working solutions, and matrix-based calibrators and QCs |
| Regulated biomarker assays | Fit-for-purpose concept; less detailed requirements | Separate section on biomarker assays; partial validation acceptable with documented scientific justification |
| Reference standard characterisation | Required; specifics varied | Explicit requirements for characterisation certificate; purity correction of nominal concentrations |
Parallelism: The LBA Requirement That Catches Teams Out
Parallelism is one of the ICH M10 requirements that most frequently requires additional work for teams transitioning from the previous FDA and EMA guidance. Parallelism testing demonstrates that the dose-response curve generated by serially diluting a study sample is parallel to the calibration curve generated from spiked matrix. Non-parallelism indicates that the analyte in study samples is behaving differently from the reference standard used to construct the calibration curve, which may be due to endogenous matrix components interfering with the assay, a different molecular form of the analyte in study samples, or binding of the analyte to carrier proteins.
Parallelism must be demonstrated during method validation using samples from the target population, not just from healthy volunteers, unless the sponsor can provide a scientific justification for why the two populations would not differ. For oncology programmes where the patient population may have elevated levels of endogenous proteins or circulating drug-related metabolites, this requirement can be practically challenging to meet before the first clinical study provides patient samples.
Reference Standard Characterisation and Purity Correction
ICH M10 introduces an explicit requirement to characterise the reference standard used in a validated bioanalytical method and to apply purity correction to the nominal concentrations of calibration standards and quality control samples. This means that if a reference standard is certified at 95% purity, the nominal concentration of a 1 nanogram per millilitre standard should be reported and calculated as 0.95 nanograms per millilitre. This sounds straightforward but has practical implications for the labelling of calibration standards in the laboratory information management system (LIMS) and for the retrospective recalculation of data from studies conducted before purity correction was applied.
Fit-for-Purpose Biomarker Assays Under ICH M10
ICH M10 formally recognises the fit-for-purpose (FFP) validation framework for biomarker assays, providing a tiered structure where the scope of validation experiments is calibrated to the intended use of the data. A biomarker assay used for exploratory characterisation in a Phase I study requires a less comprehensive validation package than the same assay used as a primary efficacy endpoint in a pivotal Phase III trial.
The ICH M10 framework requires that the validation scope for each biomarker assay is justified by the sponsor, with reference to the intended use, the decision-making consequences of the data, and the regulatory context. This justification must be documented and available for regulatory review. The table in the ICH M10 guideline that maps validation parameters to assay categories provides a practical reference for determining which experiments are required for each application.
Ardena’s ICH M10 Compliant Bioanalysis Services
Ardena’s bioanalytical laboratory in Assen operates under ICH M10-compliant method validation procedures for both chromatographic and ligand-binding assays. The standard operating procedures, validation templates, and reporting formats at the Assen facility have been updated to align with the harmonised requirements, and the scientific team is experienced in applying the parallelism, ISR, and reference standard characterisation requirements to both small molecule PK and large molecule LBA programmes.
