LC-MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine in dried blood spots

Pavan Kumar Prathipati, Subhra Mandal, Christopher J. Destache

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Abstract

A simple, short, and rugged LC-MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine was developed and validated. Dried blood spots were prepared with 25μL of spiked whole blood. A 3mm punch was extracted with methanol containing labeled internal standards. Ten microliters was injected into the LC-MS/MS using isocratic mobile phase composed of 0.1% formic acid in water and 0.1% formic acid in acetonitrile (45: 55v/v) at a flow rate of 0.25mL/min. The method was validated in the range of 10-2000ng/mL for all four analytes. The intra-assay accuracy (RE) of the method was -4.73-4.78, 1.35-2.89, -8.89 to -0.49 and-1.40-1.81 for tenofovir, emtricitabine, elvitegravir and rilpivirine, respectively. The inter-assay accuracy was within ±15% of nominal and precision (CV) was <15%. The hematocrit effect on quantification was nonsignificant at the tested hematocrit levels (35-70%). The dried blood spot method showed good agreement with the plasma method, and hence can be used as an alternative to plasma method.

Original languageEnglish (US)
JournalBiomedical Chromatography
DOIs
StateAccepted/In press - Jan 1 2018

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All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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