A validated new method for nevirapine quantitation in human plasma via high-performance liquid chromatography

Courtney F. Silverthorn, Teresa L. Parsons

Research output: Contribution to journalArticle

Abstract

A fully validated and clinically relevant assay was developed for the assessment of nevirapine concentrations in neonate blood plasma samples. Solid-phase extraction with an acid-base wash series was used to prepare subject samples for analysis. Samples were separated by high performance liquid chromatography and detected at 280 nm on a C8 reverse-phase column in an isocratic mobile phase. The retention times of nevirapine and its internal standard were 5.0 and 6.9 min, respectively. The method was validated by assessment of accuracy and precision (statistical values <15%), specificity, and stability. The assay was linear in the range 25-10,000 ng/mL (r2 > 0.996) and the average recovery was 93% (n = 18). The lower limit of quantification (relative standard deviation <20%) was determined to be 25 ng/mL for 50 μL of plasma, allowing detection of as little as 1.25 ng of nevirapine in a sample. This value represents an increase in sensitivity of up to 30-fold over previously published methods.

Original languageEnglish (US)
Pages (from-to)23-27
Number of pages5
JournalBiomedical Chromatography
Volume20
Issue number1
DOIs
StatePublished - Jan 2006

Keywords

  • HIV drug analysis
  • Nevirapine
  • Non-nucleoside reverse transcriptase inhibitor
  • Reverse-phase liquid chromatography
  • Solid-phase extraction

ASJC Scopus subject areas

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

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