Development And Validation Of Sensitive LC-MS/MS Method For Determination Of Doravirine In Human Plasma Samples

Pharmaceutical Science-Pharmaceutical analysis for Novel drug formulation

Abstract

Doravirine is a non-nucleoside reverse transcriptase inhibitor for use in the treatment of HIV/AIDS. A simple, rapid and sensitive liquid chromatography with tandem mass spectrometry (LC-MS/MS) assay method has been proposed for the determination of Doravirine in human plasma samples using Delavirdine as internal standard (IS). Analyte and the IS were extracted from the 100 µL of K2 EDTA human plasma by Solid Phase extraction (SPE). The chromatographic separation was achieved on a Zodiac C18 column by using a mixture of Methanol and 0.1%  formic acid buffer (85:15, v/v) as the mobile phase at a flow rate of 1.0 mL/min. The calibration curve obtained was linear (r2-0.99) over the concentration range of 0.15 – 40.4 ng/mL. The Mass detection of Doravirine involves m/z-426.5 (parent) and 112.5 (product) and Delavirdine involves m/z - 457.2 (parent) and 362.1 (product) as internal standard in Positive ion mode. Method validation was performed as per FDA guidelines and the results met the acceptance criteria. The intra-day and inter-day precision (%CV) and accuracy results in three validation batches across six concentration levels were well within the acceptance limits. A run time of 2.00 min for each sample made it possible to analyze  more  number  of  samples  in  short  time,  thus  increasing  the  productivity. The assay was found to be sensitive, selective and reproducible.