Bioanalysis is the process of identification, characterization, and quantification of a molecule or an analyte. It is present in different biological samples. The methods to carry out bioanalysis in a controlled environment are known as bioanalytical methods.

Bioanalytical method development and validation

Pharmaceutical companies are introducing new drugs that need approval by the appropriate authority agencies. The pharmaceutical unit must send scientifically collected data to the agencies. The data is about bioavailability, human clinical pharmacology, and bioequivalence studies. Information about non-human pharmacology and toxicology studies, along with preclinical studies, are also important. It requires a bioanalytical method which is developed and validated as per the standards and guidelines of these agencies.

EMA insight on Bioanalytical Method development and validation

FDA (Food and Drug Administration) and EMA (European Medicines Agency) are the two most popular regulatory agencies which have come up with guidelines for bioanalytical method development and validation. The FDA provides comprehensive recommendations

for reporting, while the EMA describes a precise protocol for the practical conduction of the laboratory experiments. Both these guidelines differ in their validation parameters also.

Let us have a look into the guidelines of EMA:

  • Guidelines for Method Development

It relates to the experiment design, the conditions for its operation, suitability of the method for the proposed test, its limitations, and its optimization for validation. Following points are essential for this:

  • Complete understanding of the analyte under investigation
  • Optimization of the conditions and procedures for extraction and detection of the analyte
  • Optimization of the following parameters
  • Accuracy
  • Calibration curve
  • Critical reagents
  • Minimum Required Dilution (MRD)
  • Precision
  • Quality control samples
  • Reference standards
  • Recovery
  • Selectivity
  • Sensitivity
  • Specificity
  • Stability of the analyte in the matrix

 

Guidelines for Method Validation

  • Full Validation

Full validation of a bioanalytical method is desirable when:

  • There is a need to validate the process for the quantification of the molecule of interest in clinical and specific nonclinical studies
  • Implementation of an analytical method already reported in the literature. The method is then repurposed for a commercial kit used in drug development
  • For chromatographic methods and LBAs, one must consider the following:
  • Accuracy
  • A calibration curve (response function)
  • Carry-over
  • Dilution integrity
  • Matrix effect
  • Precision
  • Range (lower limit quantification (LLOQ) and upper limit of quantification (ULOQ))
  • Reinjection reproducibility
  • Selectivity
  • Specificity
  • Stability

 

  • Partial Validation

It is necessary when a small modification occurs in a fully validated method. Extent and nature of partial validation depend on the size and type of alterations in the validated technique.

  • Cross-Validation

It is essential when the results are available from different bioanalytical methods across or within the studies, or when results are obtained from different laboratories using the same technique in a study. The comparison of these results or data needs cross-validation of the applied methods.

The EMA documents for the bioanalytical method development and validation are similar to FDA guidelines, but not identical. The International Council for Harmonization (ICH) should merge the merits of both materials to avoid the necessity to establish compliance with multiple guidelines.