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.
Aghazadeh-Habashi
Ali
A
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Sattari
Saeed
S
Pasutto
Franco
F
Jamali
Fakhreddin
F
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