Enzymatic method for the simultaneous determination of hyaluronan and chondroitin sulfates using high-performance liquid chromatography
Enzymatic method for the simultaneous determination of hyaluronan and chondroitin sulfates using high-performance liquid chromatography
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DOI：
10.1006/abio.1998.2883
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发表时间：
1998-12-01
影响因子：
2.9
通讯作者：
Imanari, T
中科院分区：
文献类型：
--
作者：
Koshiishi, I;Takenouchi, M;Imanari, T
同时测定透明质酸和硫酸软骨素的色谱方法进行了研究。在碱性条件下，Hybryonan与硫酸软骨素的不同之处在于对软骨素酶ABC的敏感性。当用软骨素酶ABC在缓冲溶液（pH 9.1）中处理透明质酸和硫酸软骨素时，硫酸软骨素被选择性地降解为不饱和二糖，而透明质酸没有。随后，在锌离子（软骨素酶ABC的抑制剂）存在下，通过链霉菌透明质酸酶在pH 6.0下将反应混合物中的透明质酸消化成不饱和四糖和六糖。通过反相离子对HPLC实现了所得不饱和二糖与硫酸软骨素和不饱和寡糖与透明质酸的分离。透明质酸和硫酸软骨素在分子量、硫酸化位置或糖醛酸组分中的结构多分散性不影响其测定。通过对猪皮样品的检测，证明了该方法的有效性。(C)北京：科学出版社.
A chromatographic method for the simultaneous determination of hyaluronan and chondroitin sulfates was examined. Hyaluronan differs from chondroitin sulfates in the susceptibility to chondroitinase ABC under alkaline conditions. When hyaluronan and chondroitin sulfates were treated with chondroitinase ABC in the buffered solution (pH 9.1), chondroitin sulfates were selectively degraded to the unsaturated disaccharides, whereas hyaluronan was not. Subsequently, hyaluronan in the reaction mixture was digested to the unsaturated tetrasaccharide and hexasaccharide at pH 6.0 by Streptomyces hyaluronidase in the presence of zinc ion (inhibitor for chondroitinase ABC). The separation of the resulting unsaturated disaccharides from chondroitin sulfates and the unsaturated oligosaccharides from hyaluronan was achieved by a reversed-phase ion-pair HPLC. The structural polydispersities of hyaluronan and chondroitin sulfates in the molecular weight, the sulfation position, or the components of uronic acid did not affect their determination. The usefulness of the present method was proved by application to the porcine skin samples. (C) 1998 Academic Press.