Liquid chromatography with electrospray ionization mass spectrometry method for the assay of glucosamine sulfate in human plasma: validation and application to a pharmacokinetic study

• Published in: Biomedical chromatography Vol. 20; no. 3; pp. 251 - 256
• Main Authors: Huang, Tao-Min, Cai, Lei, Yang, Bei, Zhou, Man-Xiang, Shen, Yun-Fei, Duan, Geng-Li
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.03.2006
• Subjects: Area Under Curve; Chromatography, Liquid - methods; electrospray ionization; Glucosamine - blood; Glucosamine - pharmacokinetics; glucosamine sulfate; Humans; mass spectrometry; Reproducibility of Results; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization - methods
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.558

A liquid chromatography–electrospray ionization mass spectrometry (LC–ESI–MS) method was developed and validated for the assay of glucosamine sulfate in human plasma. Plasma proteins were precipitated by acetonitrile, followed by vortex mixing and centrifugation. The supernatant was transferred and derivatized with phenyl iso‐thiocyanate in acetonitrile at 60°C for 40 min. Chromatographic separation was performed on a C18 column (Inertsil ODS‐3 150 × 2.1 mm i.d., 5 µm, JP) with a mobile phase gradient consisting of 0.2% acetic acid (aqueous) and methanol at a flow‐rate of 0.3 mL/min. MS detection using electrospray ionization (ESI) as an interface was used in single ion monitoring mode to determine positive ions at m/z 297. This method was shown to be selective and sensitive for glucosamine sulfate. The limit of detection was 35 ng/mL for glucosamine sulfate in plasma and the linear range was 0.1–20 µg/mL in plasma with a correlation coefficient (r) of 0.9991. The relative standard deviations (RSDs) of intra‐day and inter‐day assays were 8.7–11.4 and 9.8–12.6%, respectively. Extraction recoveries of glucosamine sulfate in plasma were greater than 73%. This method proved to be simple, reproducible and feasible for pharmacokinetic studies of glucosamine sulfate in healthy volunteers after a single oral administration (1500 mg). The pharmacokinetic parameters and relative bioavailabilities were investigated for both domestic glucosamine sulfate tablet and capsule preparations compared with an imported capsule product. Copyright © 2005 John Wiley & Sons, Ltd.