**6. Need of bioanalytical method development and validation**

The various manufacturers are applying for Investigational new drug application (IND), New drug application (NDA), Abbreviated new drug application (ANDA) to FDA. There was harmonization in this process related to human clinical pharmacology, bioavailability (BA) and Bioequivalence (BE), pharmacokinetic evaluation (PK), non-human pharmacology and toxicology studies and preclinical studies, which should be included in abovementioned applications. To obtain the data related to abovementioned requirements, there is need of development and validation of bioanalytical methods in biological matrices such as blood, serum, plasma or urine [10].

The most recent FDA guidance document on bioanalytical method development and validation was released in May 2018. Before this there was a guidance documents in 2001 and its revision in 2013 were released. The overall previous recommendations remain same, only the following points are revised.


**159**

*Analytical, Bioanalytical, Stability-Indicating Methods: Key Part of Regulatory Submissions*

• There will be the further acceptance criteria within the different batches

In this document following clarity regarding Ligand Binding Assay (LBA)

• The control of each sample should be included with clear definition

**6.1 Key principles of bioanalytical method validation and establishment**

• Accuracy, precision, selectivity, sensitivity, reproducibility and stability are the fundamental parameters that ensure the acceptability of bioanalytical

• There should be specific protocol, study plan, report or SOP for bioanalytical

• How the analyte is being get affected by environmental, matrix, or procedural variables? Every step, including time of collection of matrix and overall inves-

• The physiological nature of samples gives variable matrix. When there are Liquid Chromatography-Mass spectrometry-Mass Spectrometry (LC-MS-MS) based procedures, then protocol should be designed to avoid matrix effect,

• It is necessary to validate bioanalytical method for the intended use or

• There should be written method validation report to claim the results.

The significant change in final document, the incurred sample reanalysis section was added which includes endogenous compounds, biomarkers, diagnostic kits, bridging data and dried blood spots [11]. The guidance documents (M10) on Bioanalytical Method Development and Validation was released by International Council for Harmonization of technical requirements for pharmaceuticals for Human Use (ICH), in February 2019. Simultaneously, American association of Pharmaceutical scientists (AAPS), European bioanalysis forum (EBF), Japan Bioanalysis forum (JBF), China Bioanalysis forum (CBF) were organized a workshop of industry, academia, and health authorities to discuss this draft guidance. The objective of these discussions was the M10 guidelines which are for Bioanalytical Method Development and validation which are part of regulatory submissions. This guidance document explains the validation of Bioanalytical Methods form, which the concentration of analyte is determined from biological fluids. The concentration was obtained from pivotal nonclinical pharmacokinetic studies which are useful for taking the decisions over the regulatory submission

• The consistency in standard calibrator preparations

• The internal standard (IS) and the drift should be monitored

*DOI: http://dx.doi.org/10.5772/intechopen.93566*

• The accuracy and precision runs

including all phases of clinical trials [12].

method development and validation.

tigation time, should be clarified.

matrix may change during method validation.

method.

application.

was added


*Analytical, Bioanalytical, Stability-Indicating Methods: Key Part of Regulatory Submissions DOI: http://dx.doi.org/10.5772/intechopen.93566*


In this document following clarity regarding Ligand Binding Assay (LBA) was added

• The accuracy and precision runs

*Analytical Chemistry - Advancement, Perspectives and Applications*

**5. Bioanalytical method development and validation**

There were various regulatory agencies had done serious efforts to regulate bioanalytical method development and validation. Almost from last three decades there were large progresses in this area. The various regulatory agencies that were worked can be listed as US FDA, American association of pharmaceutical scientists (AAPS), Health protection Branch HPB, Association of analytical chemists (AOAC), Center for Veterinary medicine (CVM), U. S. Department of Health and Human Services Food and drug Administration, Center for Drug Evaluation and Research (CDER), European Medicine Agency (EMA), China Food and Drug Administration (CFDA), European Bioanalytical forum (EBF), Global CRO Council (GCC), The Brazilian health regulatory agency (ANVISA, Brazil). To regulate and harmonize bioanalytical method development and validation first workshop was held in 3–5 December 1990, report of which was published in pharmaceutical research and in other journals. On basis of the reports of this workshop, the FDA was issued draft guidance on bioanalytical method development and validation in January 1999.The second FDA guidance was published in May 2001 on the basis of workshop which was held in January 2000.The recommendations for bioanalytical method development and validation for macromolecules was published in 2006. The recommendations for regulation and harmonization of bioanalytical methods were again refreshed in 2006. In 2010, a draft guidance was published by EMA for development and validation of bioanalytical methods. As per above discussion this can be concluded that there were serious efforts carried out to regulate bioanalytical method development and validation by the various abovementioned regulatory

**6. Need of bioanalytical method development and validation**

The various manufacturers are applying for Investigational new drug application (IND), New drug application (NDA), Abbreviated new drug application (ANDA) to FDA. There was harmonization in this process related to human clinical pharmacology, bioavailability (BA) and Bioequivalence (BE), pharmacokinetic evaluation (PK), non-human pharmacology and toxicology studies and preclinical studies, which should be included in abovementioned applications. To obtain the data related to abovementioned requirements, there is need of development and validation of bioanalytical methods in biological matrices such as blood, serum,

The most recent FDA guidance document on bioanalytical method development

and validation was released in May 2018. Before this there was a guidance documents in 2001 and its revision in 2013 were released. The overall previous recom-

• There was clarification on the number of Quality control (QC) samples and

• There will be no acceptance criteria for QCs for accuracy and precision

• The QCs should have to cover the sample concentration range

• The LLOQ should be evaluated for interference for each run

mendations remain same, only the following points are revised.

• The validation criteria for dilution and carryover

**158**

agencies.

plasma or urine [10].

replicates


The significant change in final document, the incurred sample reanalysis section was added which includes endogenous compounds, biomarkers, diagnostic kits, bridging data and dried blood spots [11]. The guidance documents (M10) on Bioanalytical Method Development and Validation was released by International Council for Harmonization of technical requirements for pharmaceuticals for Human Use (ICH), in February 2019. Simultaneously, American association of Pharmaceutical scientists (AAPS), European bioanalysis forum (EBF), Japan Bioanalysis forum (JBF), China Bioanalysis forum (CBF) were organized a workshop of industry, academia, and health authorities to discuss this draft guidance. The objective of these discussions was the M10 guidelines which are for Bioanalytical Method Development and validation which are part of regulatory submissions. This guidance document explains the validation of Bioanalytical Methods form, which the concentration of analyte is determined from biological fluids. The concentration was obtained from pivotal nonclinical pharmacokinetic studies which are useful for taking the decisions over the regulatory submission including all phases of clinical trials [12].

### **6.1 Key principles of bioanalytical method validation and establishment**


There should be proper placement of the QC samples in the run.

• There is a need to set a specific acceptance criterion for bioanalytical method to be considered as a valid method. That should be achieved for accuracy and precision for validation of QC samples over the range of standards.
