Liquid chromatography–tandem mass spectrometry method for determination of N-acetylglucosamine concentration in human plasma

Abstract

A simple, reliable and sensitive liquid chromatography–tandem mass spectrometry method (LC–MS/MS) was developed and validated for quantification of N-acetylglucosamine in human plasma. Plasma samples were pretreated with acetonitrile for protein precipitation. The chromatographic separation was performed on Hypersil Silica column (150 mm × 2 mm, 5 μm). The deprotonated analyte ion was detected in negative ionization mode by multiple reaction monitoring mode. The mass transition pairs of m/ z 220.3 → 118.9 and m/ z 226.4 → 123.2 were used to detect N-acetylglucosamine and internal standard 13C 6- N-acetylglucosamine, respectively. The assay exhibited a linear range from 20 to 1280 ng/ml for N-acetylglucosamine in human plasma. Acceptable precision and accuracy were obtained for concentrations of the calibration standard and quality control. The validated method was successfully applied to analyze human plasma samples in a pharmacokinetic study.