High performance liquid chromatographic determination of glucosamine in rat plasma.
OBJECTIVE
A high performance liquid chromatographic method was developed for the determination of glucosamine (GlcN) in rat plasma.


METHODS
Internal standard, galactosamine, was added to 100 micro L of plasma containing GlcN followed by precipitation of plasma proteins with acetonitrile. Evaporation of the decanted supernatant solution was accelerated by the addition of methanol. GlcN was derivatized by addition of a solution containing 1-naphthyl isothiocyanate. Sample cleanup included passage through an anion exchange cartridge. Analysis was accomplished by injection of 0.1 mL of the sample solution into an isocratic HPLC system consisting of a C18 column, a mobile phase of acetonitrile: water: acetic acid: triethylamine (4.5: 95.5:0.1:0.05), a flow rate of 0.9 mL/min, and a UV detector set at 254 nm.


RESULTS
Galactosamine and GlcN appeared 26 and 29 min post-injection, respectively. The assay was linear over the range of 1.25-400 micro g/mL (CV<10%) with a detection limit of 0.63 microg/mL and a limit of quantification of 1.25 microg/mL. The method was applied to the determination of GlcN in rat plasma after oral administration of 350 mg/kg of GlcN hydrochloride.


CONCLUSIONS
The present assay is specific, sensitive, precise, and accurate and is suitable for pharmacokinetic studies.
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Ali
A
Aghazadeh-Habashi
Saeed
S
Sattari
Franco
F
Pasutto
Fakhreddin
F
Jamali
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005944:Glucosamine / Q000097:blood*; D008297:Male; D023421:Models, Animal; D011786:Quality Control; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards
D005944:Glucosamine