A novel ultrasensitive LC-MS/MS assay for quantification of intracellular raltegravir in human cell extracts

Brian L. Robbins, Sarah R. Nelson, Courtney V. Fletcher

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

An assay using ultrahigh pressure liquid chromatography and mass spectrometry detection was developed and validated for measurement of the HIV integrase inhibitor raltegravir (MK-0518) in human cell extracts. The assay is designed to utilize 200μl of 70% MeOH cell extract derived from human peripheral blood mononuclear cells or human tissue samples. The assay is linear over a range from 0.0023 to 9.2ngml-1. The average %CV (SD/Mean)100 and %deviation ((observed-target)/target)100 were less than 20% at the lower limit of quantification and less than 15% over the range of the curve. This assay is an accurate and highly sensitive method for determining raltegravir concentrations in cellular extracts with a lower limit 40 to over 100-fold lower than other methods in the literature. We also present a new processing method where a rapid spin through oil produced a significant increase in apparent intracellular raltegravir concentration compared with conventional processing.

Original languageEnglish (US)
Pages (from-to)378-387
Number of pages10
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume70
DOIs
StatePublished - Nov 2012

Keywords

  • HIV
  • Integrase inhibitor
  • Intracellular
  • LC-MS/MS
  • Raltegravir

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

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