TY - JOUR
T1 - Identification and quantification of glucosamine in rabbit cartilage and correlation with plasma levels by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry
AU - Pastorini, Elisabetta
AU - Vecchiotti, Stefania
AU - Colliva, Carolina
AU - Persiani, Stefano
AU - Rotini, Roberto
AU - Roatti, Giulia
AU - Zaccarelli, Lorenzo
AU - Rovati, Lucio Claudio
AU - Roda, Aldo
PY - 2011/6/10
Y1 - 2011/6/10
N2 - 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.
AB - 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.
KW - Cartilage
KW - Crystalline glucosamine sulfate
KW - Glucosamine
KW - High performance liquid chromatography-electrospray ionization-tandem mass spectrometry
KW - Knee osteoarthritis
KW - Rabbit
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U2 - 10.1016/j.aca.2011.04.003
DO - 10.1016/j.aca.2011.04.003
M3 - Article
C2 - 21601033
AN - SCOPUS:79956062559
VL - 695
SP - 77
EP - 83
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
IS - 1-2
ER -