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Improved sensitive high performance liquid chromatography assay for glucosamine in human and rat biological samples with fluorescence detection.

Abstract：

PURPOSE:An improved HPLC method with fluorescence detection was developed and validated for determination of glucosamine in human and rat biological samples. METHOD:Aliquot of 0.1 mL plasma was spiked with mannosamine HCl as the internal standard (IS); proteins were precipitated with acetonitrile; the clear layer was derivatized with 9-fluorenylmethyl chloroformate (8 mM/acetonitrile) in presence of borate 0.2 M buffer at 30 degrees C for 30 min. The excess derivatizing agent was removed with 1-aminoadamantane HCl (300 mM in acetonitrile-water 1:1). Chromatographic separation was achieved on a C18 (100 mm X 4.6 mm, id 3 microm) reversed phase column using 0.1% acetic acid/acetoniltrile gradient mobile phase at 1 mL/min flow rate. Glucosamine was determined in the plasma of a human and rats and also in rat urine. RESULTS:The analytes were detected at excitation and emission wavelengths of 263 and 315 nm, respectively. The assay was linear over the range of 0.05-20 microg/mL with a typical correlation coefficient of 0.999 and intra-day and inter-day coefficient of variation of <15%. The lowest limit of quantification was set at 50 ng/mL. The recovery for glucosamine and mannosamine was 98 and 96%, respectively. CONCLUSION:We were able to improve glucosamine assay suitable to quantify glucosamine in both human and rat plasma and rat urine.

journal_name

J Pharm Pharm Sci

journal_title

Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques

authors

Ibrahim A,Jamali F

doi

10.18433/j3t01s

subject

Has Abstract

pub_date

2010-01-01 00:00:00

pages

128-35

issue

2

issn

1482-1826

journal_volume

13

pub_type

杂志文章

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