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

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.