Laboratory Testing Blog

EMA Issues New Guideline on Bioanalytical Method Validation

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The measurement of drugs in biological matrices, such as serum, plasma, blood, urine, and saliva is an important aspect of pharmaceutical product development.  The data obtained in these clinical studies may be required to support regulatory applications for new drug products and generics as well as variations to previously approved drug products.  The results of animal toxicokinetic studies and human clinical trials, including bioequivalence studies are used to make critical decisions supporting the safety and efficacy of a drug substance or drug product. It is critical that the bioanalytical methods used are well established, fully validated and documented that will demonstrate reliable and reproducible results.

The new guideline introduced by the European Medicines Agency documents key recommendations for bioanalytical method validation applied to measure drug concentrations in biological matrices obtained in animal and human clinical studies in all phases.  This new guideline will be effective February 1, 2012.

Below are the 5 important points you will need to consider when this new guideline takes effect starting on February 1, 2012:

1. Full validation of bioanalytical methods should be performed for any bioanalytical method whether new or literature-based:

   The main objective of bioanalytical method validation is to demonstrate the reliability of a particular method for the determination of an analyte concentration in a specific biological matrix, such as blood, serum, plasma, urine, or saliva.  Generally, a full validation should be performed for each species and matrix concerned.  According to the new guideline, the main criteria to be evaluated when performing bioanalytical method validation are:

• Accuracy

• Precision

• Selectivity

• Lower Limit of Quantification (LLOQ)

• Performance of the Calibration Curve

• Matrix Effects

• Stability of the analyte(s) in the biological matrix

• Stability of the analyte(s) and of the Internal Standard in the stock and working solutions and in extracts under the entire period of storage and processing conditions

2. Separate validation recommendations for ligand binding assays will be coming into effect, since ligand binding assays are significantly different from chromatographic bioanalytical methods.

   The guideline declares that the validation principles described above and the considerations for analysis of study samples should also be applied in general for ligand-binding assays.  Due to the inherent characteristics and complex structure of these large molecules, several areas need special attention.  Over and above the general criteria used for bioanalytical method development, additional criteria to be considered include:

• Carry-over Effect

• Matrix Selection

• Minimum Required Dilution

• Dilution Linearity

• Parallelism

• Sample Stability

• Reagents

• Commercial Kits

3. The guideline stipulates when partial validation or cross-validation should be performed.  In situations where minor changes are made to a bioanalytical method that has been validated, a full validation may not be required depending on the nature of the changes.  Partial validation can range from as little as the determination of the within-run precision and accuracy to an almost full validation.  If data is obtained from different methods within and across studies or when data is obtained within a study from different bioanalytical testing laboratories, applying the same method, you will need to compare that data and a cross-validation of the bioanalytical method(s) should be performed.  The outcome of the cross-validation is critical in determining whether the data obtained is reliable and whether it can be compared and used.

4. Specific aspects of study samples analysis are addressed in the guideline.  After full bioanalytical method validation has been completed, study samples analysis can begin; but before starting the samples analysis, performance of the bioanalytical method should have been verified.  The study samples, QC samples and calibration standards should be processed in according to the validated bioanalytical method to ensure the acceptability of the study samples run.  In case a whole run consist of more batches, acceptance criteria should be applied to the whole run and to the individual batches. The run can be found acceptable, although a batch might have to be rejected, as the criteria were not met.

5. Methods used for determining quantitative concentrations of biomarkers used in assessing pharmacodynamic endpoints are not being considered in this guideline

If you would like to review the entire document, it can be found by clicking here.

The new EMA guideline, set to take effect February 1, 2012 will have far reaching impact in how bioanalytical method validation is applied and how clinical studies samples are processed and analyzed.

Are you ready for these changes? Tell us how you are preparing for it on our LinkedIn Group.