Malavaki Christina J, Asimakopoulou Athanasia P, Lamari Fotini N, Theocharis Achilleas D, Tzanakakis George N, Karamanos Nikos K
Department of Chemistry, Laboratory of Biochemistry, University of Patras, 26500 Patras, Greece.
Anal Biochem. 2008 Mar 1;374(1):213-20. doi: 10.1016/j.ab.2007.11.006. Epub 2007 Nov 9.
Exogenous administration of chondroitin sulfate (CS) is widely practiced for the treatment of osteoarthritis, although the efficacy of this treatment has not been completely established by clinical studies. A reason for the inconsistency of the results may be the quality of the CS preparations, which are commercially available as dietary supplements. In this article, we describe the development of a new method of capillary electrophoresis (CE) for the quantification of CS concentrations, screening for other glycosaminoglycan or DNA impurities and determination of hyaluronan impurities in CS raw materials, tablets, hard capsules, and liquid formulations. Analysis is performed within 12 min in bare fused silica capillaries using reversed polarity and an operating phosphate buffer of low pH. The method has high sensitivity (lower limit of quantitation [LLOQ] values of 30.0 microg/ml for CS and 5.0 microg/ml for hyaluronan), high precision, and accuracy. Analysis of 11 commercially available products showed the presence of hyaluronan impurities in most of them (up to 1.5%). CE analysis of the samples after treatment with chondroitinase ABC and ACII, which depolymerize the chains to unsaturated disaccharides, with a previously described method (Karamanos et al., J. Chromatogr. A 696 (1995) 295-305) confirmed the results of hyaluronan determination and showed that the structural characteristics (i.e., disaccharide composition) of CS are very different, showing the different species or tissue origin and possibly affecting the therapeutic outcome.
外源性给予硫酸软骨素(CS)在骨关节炎治疗中广泛应用,尽管该治疗的疗效尚未通过临床研究完全确立。结果不一致的一个原因可能是CS制剂的质量,其作为膳食补充剂在市场上销售。在本文中,我们描述了一种新的毛细管电泳(CE)方法的开发,用于定量CS浓度、筛查其他糖胺聚糖或DNA杂质以及测定CS原料、片剂、硬胶囊和液体制剂中的透明质酸杂质。使用反相极性和低pH的操作磷酸盐缓冲液,在裸露的熔融石英毛细管中12分钟内即可完成分析。该方法具有高灵敏度(CS的定量下限[LLOQ]值为30.0微克/毫升,透明质酸为5.0微克/毫升)、高精度和准确性。对11种市售产品的分析表明,其中大多数都存在透明质酸杂质(高达1.5%)。用硫酸软骨素酶ABC和ACII处理样品后进行CE分析,这些酶将链解聚为不饱和二糖,采用先前描述的方法(Karamanos等人,《色谱杂志A》696(1995)295 - 305)证实了透明质酸测定结果,并表明CS的结构特征(即二糖组成)差异很大,显示出不同的种类或组织来源,可能会影响治疗效果。