title: Development and validation of a sensitive HPLC-ESI-MS/MS method for the direct determination of glucosamine in human plasma.
abstract: A sensitive and specific HPLC-ESI-MS/MS method for the direct determination of glucosamine in human plasma has been developed and validated. Plasma samples were analyzed after a simple, one-step protein precipitation clean-up with trichloroacetic acid using a polymer-based amino high-performance liquid chromatography (HPLC) column and a water/acetonitrile mobile phase elution gradient, with d-[1-(13)C]glucosamine as the internal standard. Detection was performed by mass spectrometry, using an electrospray source and employing multiple reaction monitoring to separately monitor glucosamine and the internal standard. The limit of quantification of the method was 10ng/ml of glucosamine and the calibration curve showed a good linearity up to 1000ng/ml. The precision (R.S.D.) and the accuracy (bias) of the method at the limit of quantification were 13.8 and 4.0%, respectively, and the mean recovery of glucosamine at three concentration levels was 101.6+/-5.7%. The method was applied for the determination of glucosamine concentrations in human plasma samples collected from untreated healthy volunteers and, in a separate bioavailability study, to evaluate plasma glucosamine pharmacokinetics profiles after oral administration of crystalline glucosamine sulfate.
authors.affiliation: Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, I-40126 Bologna, Italy. aldo.rods@unibo.it
authors.forename: Aldo
authors.initials: A
authors.lastname: Roda
authors.forename: Laura
authors.initials: L
authors.lastname: Sabatini
authors.forename: Anna
authors.initials: A
authors.lastname: Barbieri
authors.forename: Massimo
authors.initials: M
authors.lastname: Guardigli
authors.forename: Marcello
authors.initials: M
authors.lastname: Locatelli
authors.forename: Francesco Saverio
authors.initials: FS
authors.lastname: Violante
authors.forename: Lucio C
authors.initials: LC
authors.lastname: Rovati
authors.forename: Stefano
authors.initials: S
authors.lastname: Persiani
mesh_terms: D001682:Biological Availability; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005944:Glucosamine / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D005944:Glucosamine