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Abstract

BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS DETERMINATION OF BICTEGRAVIR, TENOFOVIR AND EMTRICITABINE IN HUMAN PLASMA BY LC-MS/MS

Dr. VVSS. Appala Raju, Dr. Macharla Venkata Ramana, Dr. T. Shyam. Principal, K.N. Rajinikanth, Dr. V. Kiran Kumar and *Dr. N. Appala Raju

ABSTRACT

A simple, sensitive and fast throughput liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed for the simultaneous estimation of bictegravir, tenofovir and emtricitabine in human plasma, using respective didanosine, stavudine and abacavir as internal standards respectively. The method involved Liquid-Liquid Extraction of the analytes and internal standards from human plasma. The chromatographic separation was achieved on a Zorbax C18 column (150×4.6mm and 5μm particle size) analytical column using isocratic mobile phase, consisting of Methanol : 0.1% formic acid in water (85:15, v/v), at a flow-rate of 1.0 mL/min with 90% flow splitting. The parent→product ion transitions were monitored at m/z 268.2 → 127.1 (BTGR), m/z 237.1→137.1 (DDI), m/z 230.2→112.1 (TNFR), m/z 248.1→130.0 (D4T), m/z 267.2→226.1 (EMTB) and m/z 287.2→191.2 (ABC) on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) positive ion mode. The method was validated over the concentration range of 5-1500 ng for Bictegravir and tenofovir and 10-3000 ng/mL for Emtricitabine. The mean recovery values for both the drugs from spiked plasma samples were reproducible. The method was rugged and rapid with a total run time of 4.0 minutes.

Keywords: Bictegravir, Tenofovir and Emtricitabine, LC-MS, Extraction.


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