Determination of a depolymerized holothurian glycosaminoglycan in plasma after intravenous administration by postcolumn HPLC.
Depolymerized holothurian glycosaminoglycan (DHG) produced artificially from sea cucumber has an anticoagulant and antithrombotic activity. In the present study, we attempted to determine the plasma level of DHG using a chemical procedure. A general method for determination of chondroitin sulfates by forming unsaturated disaccharides with chondroitinase digestion was difficult to apply to DHG, because it was resistant to any chondroitinase digestion. We therefore developed a highly sensitive postcolumn HPLC method for determination of intact DHG. DHG was applied to Asahipak NH2P-50, an amino-bonded column, eluted by alkaline solution and then detected with arginine under a strong alkaline condition as a postcolumn reagent. The limit of detection for DHG was 10 ng. The present method was applicable to the determination of DHG in plasma after intravenous administration.
Huang
Y
Y
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Toyoda
H
H
Koshiishi
I
I
Toida
T
T
Imanari
T
T
D000818:Animals; D000925:Anticoagulants / Q000008:administration & dosage / Q000097:blood* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D005343:Fibrinolytic Agents / Q000008:administration & dosage / Q000097:blood* / Q000302:isolation & purification; D006025:Glycosaminoglycans / Q000008:administration & dosage / Q000097:blood* / Q000302:isolation & purification; D007275:Injections, Intravenous; D008297:Male; D011817:Rabbits; D012616:Sea Cucumbers / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013696:Temperature
D000925:Anticoagulants; D005343:Fibrinolytic Agents; D006025:Glycosaminoglycans