Identification and quantification of glucosamine in rabbit cartilage and correlation with plasma levels by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry

Elisabetta Pastorini, Stefania Vecchiotti, Carolina Colliva, Stefano Persiani, Roberto Rotini, Giulia Roatti, Lorenzo Zaccarelli, Lucio Claudio Rovati, Aldo Roda

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A new HPLC-ESI-MS/MS method for the determination of glucosamine (2-amino-2-deoxy-d-glucose) in rabbit cartilage was developed and optimized. Glucosamine was extracted from cartilage by cryogenic grinding followed by protein precipitation with trichloroacetic acid. The HPLC separation was achieved with a polymer-based amino column using a mobile phase composed of 10mM ammonium acetate (pH 7.5)-acetonitrile (20:80%, v/v) at 0.3mLmin -1 flow rate. d-[1- 13C]Glucosamine was used as internal standard. Selective detection was performed by tandem mass spectrometry with electrospray source, operating in positive ionization mode and in multiple reaction monitoring acquisition (m/z 180→72 and 181→73 for glucosamine and internal standard, respectively). Limit of quantification was 0.045ng injected, corresponding to 0.25μgg -1 in cartilage. Linearity was obtained up to 20μgg -1 (R 2>0.991). Precision values (%R.S.D.) were -1 (n=6). Glucosamine was present in cartilage in physiological condition before the treatment. After dosing, mean concentration of cartilage glucosamine significantly increased from 461 to 1040ngg -1. Cartilage glucosamine levels resulted to be well correlated with plasma concentrations, which therefore are useful to predict the target cartilage concentration and its pharmacological activity.

Original languageEnglish
Pages (from-to)77-83
Number of pages7
JournalAnalytica Chimica Acta
Volume695
Issue number1-2
DOIs
Publication statusPublished - Jun 10 2011

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Electrospray ionization
Electrospray Ionization Mass Spectrometry
Glucosamine
Cartilage
High performance liquid chromatography
Tandem Mass Spectrometry
Mass spectrometry
liquid chromatography
ionization
mass spectrometry
High Pressure Liquid Chromatography
Rabbits
Plasmas
plasma
linearity
grinding
acetate
glucose
ammonium
polymer

Keywords

  • Cartilage
  • Crystalline glucosamine sulfate
  • Glucosamine
  • High performance liquid chromatography-electrospray ionization-tandem mass spectrometry
  • Knee osteoarthritis
  • Rabbit

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Spectroscopy
  • Environmental Chemistry

Cite this

Identification and quantification of glucosamine in rabbit cartilage and correlation with plasma levels by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry. / Pastorini, Elisabetta; Vecchiotti, Stefania; Colliva, Carolina; Persiani, Stefano; Rotini, Roberto; Roatti, Giulia; Zaccarelli, Lorenzo; Rovati, Lucio Claudio; Roda, Aldo.

In: Analytica Chimica Acta, Vol. 695, No. 1-2, 10.06.2011, p. 77-83.

Research output: Contribution to journalArticle

Pastorini, Elisabetta ; Vecchiotti, Stefania ; Colliva, Carolina ; Persiani, Stefano ; Rotini, Roberto ; Roatti, Giulia ; Zaccarelli, Lorenzo ; Rovati, Lucio Claudio ; Roda, Aldo. / Identification and quantification of glucosamine in rabbit cartilage and correlation with plasma levels by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry. In: Analytica Chimica Acta. 2011 ; Vol. 695, No. 1-2. pp. 77-83.
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