Purification of heparin, dermatan sulfate and chondroitin sulfate from mixtures by sequential precipitation with various organic solvents.
Heparin, dermatan sulfate and chondroitin sulfate in mixtures were fractionated by sequential precipitation with methanol, ethanol and propanol. The recovered fractions from 0.1 to 2.0 volumes of various solvents were analyzed by agarose-gel electrophoresis and densitometric analysis. Heparins with different relative percentages of slow-moving and fast-moving components were precipitated from 0.5 to 0.7 volumes of methanol, and in this range of volumes, the amount of slow-moving component of heparin decreases and that of the fast-moving species increases. From 0.8 to 1.6 volumes of methanol, mixtures with different percentages of the fast-moving component, dermatan sulfate and chondroitin sulfate are precipitated. Heparin was precipitated from mixtures in the range of 0.1 to 0.4 volumes of ethanol, and from 0.5 to 0.8 volumes mixtures with different relative percentages of dermatan sulfate and chondroitin sulfate were precipitated. From 1.0 to 2.0 volumes of ethanol, high purity (about 100%) chondroitin sulfate can be precipitated. Propanol induces the precipitation of heparin from 0.3 to 0.4 volumes, whilst dermatan sulfate with a purity greater than 85% is precipitated at 0.5 and 0.6 volumes of propanol. 100% chondroitin sulfate is obtained with volumes greater than 0.8. Heparin and chondroitin sulfate from a bovine lung extract of glycosaminoglycans were purified by sequential precipitation with ethanol. The fraction precipitated with 0.4 volumes of ethanol shows greater than 90% heparin and that recovered from 0.9 to 2.0 volumes is composed of 100% chondroitin sulfate.
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Department of Biologia Animále, University of Modena, Italy.
D000818:Animals; D002417:Cattle; D002809:Chondroitin Sulfates / Q000302:isolation & purification*; D003720:Densitometry; D003871:Dermatan Sulfate / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D006493:Heparin / Q000302:isolation & purification*; D008168:Lung / Q000737:chemistry; D012997:Solvents
D012997:Solvents; D003871:Dermatan Sulfate; D006493:Heparin; D002809:Chondroitin Sulfates