A Liquid Chromatography Tandem Mass Spectrometry Based Regulatory Compliant Method for the Determination of Tenofovir in Human Serum

Abstract

A simple, rapid, and specific assay based on solid phase extraction and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI MS-MS) for the quantitative analysis of Tenofovir in human serum using Tenofovir D6 as internal standard (IS) have been developed. The precursor to product ion transitions of m/z 288.2/176.2 and m/z 293.9/182.3 used to measure the analyte and internal standard (Tenofovir and Tenofovir D6). The method was validated over a concentration range of 5.06–603.72 ng mL⁻¹. The method was validated over the parameters like selectivity, matrix effect, sensitivity, linearity, precision, accuracy, various stabilities (bench top stability, standard stock solution stability in refrigerator and at room temperature, stock dilution stability, auto sampler stability, freeze thaw stability, long term stability – 65°C±10°C & long term stability – 22°C±5°C, reagent stability, dry extract stability, wet extract stability in refrigerator and at bench top, blood stability), effect of potentially interfering drugs, dilution integrity, recovery and reinjection reproducibility. The mean % recovery of Tenofovir was 98.22% with a precision of 2.42%, The mean % recovery of Tenofovir D6 was 100.96% with a precision of 2.88%. The RSD % of intra-day and inter-day assay was≤15%.

Key words

Tenofovir - electrospray ionization - tandem mass spectrometry - human serum - bioequivalence study

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