Analytical Biochemistry 448 (2014) 113–115
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Analytical Biochemistry journal homepage: www.elsevier.com/locate/yabio
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Selective removal of keratan sulfate in chondroitin sulfate samples by sequential precipitation with ethanol Fabio Galeotti, Francesca Maccari, Nicola Volpi ⇑ Department of Life Sciences, University of Modena and Reggio Emilia, 41100 Modena, Italy
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Article history: Received 11 October 2013 Received in revised form 29 October 2013 Accepted 22 November 2013 Available online 1 December 2013 Keywords: Chondroitin sulfate Keratan sulfate Ethanol precipitation Hexosamines Capillary electrophoresis Glycosaminoglycans
a b s t r a c t Keratan sulfate (KS) is present as a contaminant in chondroitin sulfate (CS) mainly extracted from shark cartilage. We report a selective removal procedure of KS in CS samples by means of sequential precipitation with ethanol. Purified shark CS containing approximately 10% to 15% KS was subjected to a precipitation procedure in the presence of increasing percentages of saturated ethanol. In contrast to other solvents, 1.0 volume of ethanol was able to selectively purify CS, with a purity of approximately 100%, from KS. The current selective and simple procedure appears to be a reliable industrial preparation of CS devoid of large amounts of the residual KS. Ó 2013 Elsevier Inc. All rights reserved.
Chondroitin sulfate (CS)1, a heteropolysaccharide belonging to the class of glycosaminoglycans (GAGs), is an oral-administered SYSADOA (symptomatic, slow-acting drug for the treatment of osteoarthritis) able to provide pain relief and increased mobility in osteoarthritis patients [1]. Moreover, recent clinical trials have demonstrated the capacity of CS as a structure-modifying agent able to act as a DiMoDOA (disease-modifying drug in osteoarthritis) [2]. In addition, CS alone or in combination with glucosamine and/or other ingredients is used as a nutraceutical in Europe, the United States, Australia, and other countries [3]. CS is extracted and purified from animal sources, and its commercial manufacture mainly relies on bovine, porcine, and chicken cartilages, cartilaginous fishes such as shark and skate, and their combination [4,5]. As a consequence, depending on the purification protocols, the presence of contaminants and polluting side products contributes to the overall pharmacological, nutraceutical, and biological action of this macromolecule. Keratan sulfate (KS) has been detected in many batches of CS extracted from shark cartilage, averaging approximately 16% of the total GAGs [6]. On the contrary, KS content was found to be very low (
Contents lists available at ScienceDirect
Analytical Biochemistry journal homepage: www.elsevier.com/locate/yabio
Notes & Tips
Selective removal of keratan sulfate in chondroitin sulfate samples by sequential precipitation with ethanol Fabio Galeotti, Francesca Maccari, Nicola Volpi ⇑ Department of Life Sciences, University of Modena and Reggio Emilia, 41100 Modena, Italy
a r t i c l e
i n f o
Article history: Received 11 October 2013 Received in revised form 29 October 2013 Accepted 22 November 2013 Available online 1 December 2013 Keywords: Chondroitin sulfate Keratan sulfate Ethanol precipitation Hexosamines Capillary electrophoresis Glycosaminoglycans
a b s t r a c t Keratan sulfate (KS) is present as a contaminant in chondroitin sulfate (CS) mainly extracted from shark cartilage. We report a selective removal procedure of KS in CS samples by means of sequential precipitation with ethanol. Purified shark CS containing approximately 10% to 15% KS was subjected to a precipitation procedure in the presence of increasing percentages of saturated ethanol. In contrast to other solvents, 1.0 volume of ethanol was able to selectively purify CS, with a purity of approximately 100%, from KS. The current selective and simple procedure appears to be a reliable industrial preparation of CS devoid of large amounts of the residual KS. Ó 2013 Elsevier Inc. All rights reserved.
Chondroitin sulfate (CS)1, a heteropolysaccharide belonging to the class of glycosaminoglycans (GAGs), is an oral-administered SYSADOA (symptomatic, slow-acting drug for the treatment of osteoarthritis) able to provide pain relief and increased mobility in osteoarthritis patients [1]. Moreover, recent clinical trials have demonstrated the capacity of CS as a structure-modifying agent able to act as a DiMoDOA (disease-modifying drug in osteoarthritis) [2]. In addition, CS alone or in combination with glucosamine and/or other ingredients is used as a nutraceutical in Europe, the United States, Australia, and other countries [3]. CS is extracted and purified from animal sources, and its commercial manufacture mainly relies on bovine, porcine, and chicken cartilages, cartilaginous fishes such as shark and skate, and their combination [4,5]. As a consequence, depending on the purification protocols, the presence of contaminants and polluting side products contributes to the overall pharmacological, nutraceutical, and biological action of this macromolecule. Keratan sulfate (KS) has been detected in many batches of CS extracted from shark cartilage, averaging approximately 16% of the total GAGs [6]. On the contrary, KS content was found to be very low (
