Author(s): Sambasiva Naik Nunsavathu, B. Thangabalan


DOI: 10.52711/2231-5675.2024.00009   

Address: Sambasiva Naik Nunsavathu*, B. Thangabalan
Department of Pharmaceutical Analysis, SIMS College of Pharmacy, Guntur, Andhrapradesh - 522002.
*Corresponding Author

Published In:   Volume - 14,      Issue - 1,     Year - 2024

Bioanalytical method development is the process of creating a procedure to enable a compound of interest to be identified and quantified in a biological matrix. A compound can often be measured by several methods and the choice of analytical method involves many considerations. Analysis of drugs and their metabolites in a biological matrix is carried out using different extraction techniques like liquid-liquid extraction, solid phase extraction (SPE) and protein precipitation from these extraction methods samples are spiked with calibration (reference) standards and using quality control (QC) samples. These methods and choice of analytical method describes the process of method development and includes sampling, sample preparation, separation, detection and evaluation of the results. The developed process is then validated. These bioanalytical validations play a significant role in evaluation and interpretation of bioavailability, bioequivalence, pharmacokinetic, and toxicokinetic studies. In which different parameters like accuracy, precision, selectivity, sensitivity, reproducibility, and stability are performed.

Cite this article:
Sambasiva Naik Nunsavathu, B. Thangabalan. A RP-HPLC approach to Bioanalytical Method Development and Validation: A Review. Asian Journal of Pharmaceutical Analysis. 2024; 14(1):47-0. doi: 10.52711/2231-5675.2024.00009

Sambasiva Naik Nunsavathu, B. Thangabalan. A RP-HPLC approach to Bioanalytical Method Development and Validation: A Review. Asian Journal of Pharmaceutical Analysis. 2024; 14(1):47-0. doi: 10.52711/2231-5675.2024.00009   Available on:

1.    D. Zimmer, Introduction to quantitative liquid chromatography-tandem mass spectrometry (LC-MS-MS), Chromatographia. 2003; 57: 325–332.
2.    J. Schuhmacher, D. Zimmer, F. Tesche, V. Pickard, Matrix effects during analysis of plasma samples by electrospray and atmospheric pressure chemical ionization mass spectrometry: practical approaches to their elimination, Rapid Commun. Mass Spectrom. 2003; 17: 1950–1957.
3.    E. Stokvis, H. Rosing, J.H. Beijnen, Stable isotopically labeled internal standards in quantitative bioanalysis using liquid chromatography/mass spectrometry: necessity or not?, Rap. Com. Mass Spec. 2005; 19: 401–407.
4.    T.M. Annesley, Ion suppression in mass spectrometry, Clin. Chem. 2003; 7 : 1041–1047.
5.    B.K. Matuszewski, M.L Constanzer, C.M. Chavez-Eng, Strategies for the Assessment of Matrix Effect in Quantitative Bioanalytical Methods Based on HPLC−MS/MS, Anal. Chem. 2003; 75: 3019–3030.
6.    R. Weaver, R.J. Riley, Identification and reduction of ion suppression effects on pharmacokinetic parameters by polyethylene glycol 400, Rap. Com. Mass Spec. 2006; 20: 2559–2564.
7.    A. V. Eeckhaut, K. Lanckmans, S. Sarre, I. Smolders, Y. Michotte, Validation of bioanalytical LC–MS/MS assays: Evaluation of matrix effects, J. Chromatogr. B. 2009; 877: 2198–2207.
8.    M. Ahnoff, A-C. Nyström, F. Schweikart, A. Ekdahl, Matrix effect explained by unexpected formation of peptide in acidified plasma, Bioanalysis. 2015; 7: 295-306
9.    [12] M.W.J. van Hout, H.A.G. Niederländer, R.A. de Zeeuw, G.J. de Jong, Ion suppression in the determination of clenbuterol in urine by solid-phase extraction atmospheric pressure chemical ionisation ion-trap mass spectrometry, Rap. Com. Mass Spec.2003; 17: 245–250.
10.    D. Remane, M.R. Meyer, F.T. Peters, D.K. Wissenbach, H.H. Maurer, Fast and simple procedure for liquid–liquid extraction of 136 analytes from different drug classes for development of a liquid chromatographic tandem mass spectrometric quantification method in human blood plasma, Anal. Bioanal. Chem. 2010; 397: 2303–2314.
11.    O. González, M.E. Blanco, G. Iriarte, L. Bartolome, M.I. Maguregui, R.M. Alonso, Bioanalytical chromatographic method validation according to current regulations, with a special focus on the non-well defined parameters limit of quantification, robustness and matrix effect, J. Chromatogr. A. 2014; 1353: 10–27.
12.    A.R. Buick, M.V. Doig, S.C. Jeal, G.S. Land and R.D. McDowall, Method Validation in the Bioanalytical Laboratory, J. Pharm. Biomed. Anal. 1990; 8: 629–637.
13.    Mohamed Abdel-Rehim, Bioanalytical method development and validation: critical concepts and strategies,2016
14.    S. Kamaruzaman, M.M. Sanagi, S. Endud, W.A. Wan Ibrahim, N. Yahaya, MCM-41 solid phase membrane tip extraction combined with liquid chromatography for the determination of non-steroidal anti-inflammatory drugs in human urine, J. Chromatogr. B. 2013; 940: 59–65.
15.    G.M. Ben-Hander, A. Makahleh, B. Saad, M.I. Saleh, Hollow fiber liquid phase microextraction with in situ derivatization for the determination of trace amounts of metformin hydrochloride (anti-diabetic drug) in biological fluids, J. Chromatogr. B. 2013; 941: 123–130.

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DOI: 10.5958/2231–5675 

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