Analysis of aflatoxin B1-lysine adduct in serum using isotope-dilution liquid chromatography/tandem mass spectrometry

Leslie F. McCoy, Peter F. Scholl, Rosemary L. Schleicher, John D. Groopman, Carissa D. Powers, Christine M. Pfeiffer

Research output: Contribution to journalArticle

Abstract

A method for quantitative analysis of aflatoxin B1-lysine adduct (B1-Lys) in serum by liquid chromatography using tandem mass spectrometry (LC/MS/MS) is presented. The protein in a 250-μL sample was digested in the presence of a stable-isotope internal standard during a 4-h incubation at 37°C with Pronase™. B1-Lys and the internal standard were extracted using mixed-mode solid-phase extraction cartridges and eluted with 2% formic acid in methanol. Following evaporation and reconstitution, extracts were injected onto a Luna C-18(2) column and eluted with a step gradient of acetonitrile and 0.06% formic acid. The B1-Lys and the internal standard were detected in a positive ionization selective reaction monitoring mode with a ThermoFinnigan TSQ Quantum triple quadrupole mass spectrometer. Calibration curves were linear for concentrations from 0.05-8.0 ng/mL. The method was validated with aflatoxin B1 dosed rat serum diluted to anticipated high and low concentrations. Total imprecision determined from 30 measurements over 15 days was 5.6% and 9.1%, respectively. Recoveries of 78.8 ± 6.4% for B1-Lys and 85.4 ± 12.4% for the internal standard were based on the full extraction and reconstitution processes. The method can be used to quantitate B1-Lys at the 0.5 pg/mg albumin level and is suitable for routine analysis.

Original languageEnglish (US)
Pages (from-to)2203-2210
Number of pages8
JournalRapid Communications in Mass Spectrometry
Volume19
Issue number16
DOIs
StatePublished - Aug 29 2005

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Organic Chemistry

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