Pretreatment Procedure for the Microdetermination of Chondroitin Sulfate in Plasma and Urine

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Authors
Sakai, Shinobu
Onose, Jun Ichi
Nakamura, Haruka
Toyoda, Hidenao
Toida, Toshihiko
Imanari, Toshio
Linhardt, Robert J.
Issue Date
2002-03-15
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Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Abstract
A new, simple, and rapid pretreatment method for the determination of chondroitin sulfate, dermatan sulfate, and hyaluronan from urine and blood plasma samples has been developed. Plasma proteins were first converted into small peptides by digestion using a nonspecific protease, actinase E, and the resulting small peptides were removed by centrifugal filtration. The retained, residual crude glycosaminoglycans, including chondroitin/dermatan sulfates and hyaluronan, were converted into unsaturated disaccharides through the action of chondroitin sulfate lyses. Next, these disaccharides were recovered and purified using centrifugal filtration together with DeltaDi-UA2S, added as an internal standard. The filtered disaccharide mixture was analyzed by HPLC with fluorometric postcolumn derivatization using 2-cyanoacetamide as a fluorogenic reagent. This method was applied to a pharmacokinetic study of chondroitin sulfate administered intravenously to mice. The half-life of the administered chondroitin sulfates, having molecular masses from 6 to 50 kDa, varied depending on their molecular sizes. This new method should be useful for studies on the metabolic fate of exogenously administered glycosaminoglycans in small experimental animals.
Description
Analytical Biochemistry, 302, 169-174
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Full Citation
Pretreatment Procedure for the Microdetermination of Chondroitin Sulfate in Plasma and Urine, S. Sakai, J. Onose, H. Nakamura, H. Toyoda, T. Toida, Toshio Imanari, R. J. Linhardt, Analytical Biochemistry, 302, 169-174, 2002.
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32697
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