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DOI: 10.1055/s-0033-1361090
A Liquid Chromatography Tandem Mass Spectrometry Based Regulatory Compliant Method for the Determination of Tenofovir in Human Serum
Publication History
received 14 May 2012
accepted 25 September 2013
Publication Date:
05 December 2013 (online)

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%.
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References
- 1 Chapman T, McGavin J, Noble S. Tenofovir disoproxil fumarate. Drugs 2003; 63: 1597-1608
- 2 Hazra R, Balis FM, Tullio AN et al. Single-Dose and Steady-State Pharmacokinetics of Tenofovir Disoproxil Fumarate in Human Immunodeficiency Virus-Infected Children, Antimicrob. Agents Chemother 2004; 48: 124
- 3 Dando Toni M, Wagstaff Antona J. Emtricitabine/Tenofovir Disoproxil Fumarate, Drugs 2004; 64: 2075-2082
- 4 http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm081336.htm
- 5 Guo J, Meng F, Li L et al. Development and validation of an LC/MS/MS method for the determination of tenofovir in monkey plasma. Biol Pharm Bull 2011; 34: 877-882
- 6 Gomes NA, Vaidya VV, Pudage A et al. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of tenofovir and emtricitabine in human plasma and its application to a bioequivalence study. J Pharm Biomed Anal 2008; 48: 918-926 Epub 2008 Aug 5
- 7 Delahunty T, Bushman L, Robbins B et al. The simultaneous assay of tenofovir and emtricitabine in plasma using LC/MS/MS and isotopically labeled internal standards. J Chromatogr B Analyt Technol Biomed Life Sci 2009; 877: 1907-1914
- 8 Yadav M, Singhal P, Goswami S et al. Selective determination of antiretroviral agents tenofovir, emtricitabine, and lamivudine in human plasma by a LC-MS-MS method for a bioequivalence study in healthy Indian subjects. J Chromatogr Sci 2010; 48: 704-713
- 9 Bennetto-Hood C, Long MC, Acosta EP. Development of a sensitive and specific liquid chromatography/mass spectrometry method for the determination of tenofovir in human plasma. Rapid Commun Mass Spectrom 2007; 21: 2087-2094
- 10 Takahashi M, Kudaka Y, Okumura N et al. Determination of plasma tenofovir concentrations using a conventional LC-MS method. Biol Pharm Bull 2007; 30: 1784-1786