Development and validation of a LC-MS/MS method for the quantification of tenofovir and emtricitabine in seminal plasma

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Oct 15;1033-1034:234-241. doi: 10.1016/j.jchromb.2016.08.011. Epub 2016 Aug 11.


Accurate and sensitive liquid-chromatography tandem mass spectrometry method for the quantification of tenofovir and emtricitabine in seminal plasma has been developed and full validated. Molecules were separated by high-performance liquid chromatography on an Atlantis T3 C18 column using a gradient of deionized water and methanol, including 0.05% formic acid (250μl/min) and detected by electrospray ionisation/tandem mass spectrometry in positive ion mode. The method was validated over a clinical range of 3.13-1000ng/mL for tenofovir and 6.25-2000ng/mL for emtricitabine. Inter and intra-assay precisions were <9.37% for tenofovir and<10.88% for emtricitabine, and accuracies were between 0.48% and 8.43% for tenofovir, and between 0.64% and 13.87% for emtricitabine. The developed method was successfully applied for analysing tenofovir and emtricitabine concentrations in seminal plasma samples from a clinical study. The use of tandem mass spectrometry can be a suitable method for the analysis of this kind of matrices, providing high sensitivity and specificity to the analysis.

Keywords: Emtricitabine; LC–MS/MS; Seminal plasma; Tenofovir.

Publication types

  • Validation Study

MeSH terms

  • Chromatography, Liquid / methods*
  • Drug Stability
  • Emtricitabine / analysis*
  • Emtricitabine / chemistry
  • Humans
  • Limit of Detection
  • Male
  • Reference Standards
  • Reproducibility of Results
  • Semen / chemistry*
  • Tandem Mass Spectrometry / methods*
  • Tenofovir / analysis*
  • Tenofovir / chemistry


  • Tenofovir
  • Emtricitabine