Development and Validation of a New HPLC Bio Analytical Internal Standard Method for the Analysis of Two Immunology Drugs (Lamivudine and Dolutegravir) in Human Plasma
DOI:
https://doi.org/10.61841/529zhk75Keywords:
RP-HPLC, Internal Standard, Bio-analytical, ICH Guidelines, Lamivudine, Dolutegravir and Abacavir.Abstract
Bioanalytical methods are useful for the quantitative analysis of drugs and their metabolites in
biological fluids. Bio-analytical internal standards methods plays vital role in areas of human clinical
pharmacology. The aim of this work is to development and validation of bio-analytical internal standard method for
determination of two immunology drugs Lamivudine (LMVD) and Dolutegravir (DTGR) in plasma with Abacavir
(ABVR) drug as internal standard (IS). Liquid-liquid extraction with Diethyl ether and methanol in the ratio of
50:50 (v/v) was used for the extraction of drugs from the biological matrix. The optimized chromatography
conditions consist of Water, Acetonitrile and Methanol in the ratio of 40:25:35 (v/v) as a mobile phase with
Ascentis, C18 Column (250 X 4.6 mm, 5μ) as stationery phase. Isocratic elution with 1.2 ml flow separates the
LMVD at 3.5 min, DTGR at 9.8 min and ABVR at 6.8 min. The method was validated as per ICH guidelines. The
relative standard deviation (%RSD) were found to be <5%for precision studies. Hence the method was found to be
suitable fort the analysis of LMVD and DTGR in spiked human plasma.
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