title: Validation of an UPLC-MS/MS method for quantitative analysis of raltegravir in human plasma samples.
abstract: BACKGROUND
  An ultra-performance liquid chromatography-tandem mass spectrometry method was developed for the quantification of raltegravir (RTG) plasma concentrations in samples from HIV patients treated with the drug.
  
  
  METHODS
  Plasma samples were extracted by liquid-liquid extraction followed by evaporation to dryness and reconstitution in mobile phase. The chromatographic separation was carried out on an AQUITY UPLC C18 column with an isocratic mobile phase consisting of water containing 0.1% formic acid and acetonitrile containing 0.1% formic acid (50:50 vol/vol). The detection was performed on a triple quadrupole tandem mass spectrometer using multi-reaction monitoring via electrospray ionization source with positive ionization mode.
  
  
  RESULTS
  Under these conditions, a single chromatographic run could be completed within 1 minute. The method was validated by estimating the precision and the accuracy for inter- and intra-day analysis in the concentration range of 5-2560 ng/mL. The method was linear over the investigated range with all the correlation coefficients, r, greater than 0.995 on 5 replicates. The intra- and inter-day precision (percentage of coefficient of variation) ranged from 2.4% to 11.2%, and the inaccuracy (percent of relative standard deviation) ranged from 2.5% to 12.9%. No significant matrix effect was observed. The mean recovery value of RTG was 80%.
  
  
  CONCLUSIONS
  This rapid and sensitive method was validated and could be applied to pharmacokinetic studies for the determination of RTG concentrations in human plasma samples.
authors: Clinical Pharmacology Unit, Agostino Gemelli Academic Hospital, Rome, Italy.
authors: Serena
authors: S
authors: Fortuna
authors: Enzo
authors: E
authors: Ragazzoni
authors: Lucia
authors: L
authors: Lisi
authors: Simona
authors: S
authors: Di Giambenedetto
authors: Massimiliano
authors: M
authors: Fabbiani
authors: Pierluigi
authors: P
authors: Navarra
mesh_terms: D015658:HIV Infections / Q000097:blood; D006801:Humans; D059625:Liquid-Liquid Extraction / Q000379:methods / Q000592:standards*; D011760:Pyrrolidinones / Q000097:blood*; D000068898:Raltegravir Potassium; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000592:standards*
chemical_list: D011760:Pyrrolidinones; D000068898:Raltegravir Potassium