Determination of glucosamine sulfate in human plasma by precolumn derivatization using high performance liquid chromatography with fluorescence detection: its application to a bioequivalence study.
A simple, rapid, selective and specific high-performance liquid chromatography (HPLC) method with fluorescence detection was developed for determination of glucosamine sulfate in human plasma and application to a bioequivalence in healthy volunteers. Precipitation of plasma was accomplished with acetonitrile to separate interfering endogenous products from the compound of interest. After vortex mixing and centrifugation, the supernatant was transferred and derivatized with 9-fluorenylmethoxycarbonyl chloride-acetonitrile solution in borate buffer (pH=8.0) at 30 degrees C for 30 min. The chromatographic separation was performed on a Diamonsil C18 column (150.0 mmx4.6 mm, 5 microm) with a mobile phase gradient consisting of water and acetonitrile at a flow rate of 1 mL/min. The method was linear in the range of 0.1-10.0 microg/mL with a correlation coefficient (r) of 0.9996. The limit of detection was 15 ng/mL. Inter- and intra-day precisions were <or=6.28 and 7.41%, respectively, and the accuracy ranged from 95.20 to 104.92%. Extraction recoveries of glucosamine sulfate from plasma were more than 90.4%. Plasma samples containing glucosamine sulfate were stable for 40 days at -20 degrees C and for 24 h after derivatization at 4 degrees C. The method was successfully applied to the bioequivalence study of glucosamine sulfate in healthy volunteers.
Zhang
Li-jun
LJ
Department of Pharmaceutics, School of Pharmacy, Fudan University, and Department of Eye & ENT Hospital, Shanghai 200032, PR China.
Huang
Tao-min
TM
Fang
Xiao-ling
XL
Li
Xue-ning
XN
Wang
Qing-song
QS
Zhang
Zhi-wen
ZW
Sha
Xian-yi
XY
D000284:Administration, Oral; D002681:China; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005944:Glucosamine / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013810:Therapeutic Equivalency
D005944:Glucosamine