Quantitative capillary electrophoresis determination of oversulfated chondroitin sulfate as a contaminant in heparin preparations.
Quantitative capillary electrophoresis determination of oversulfated chondroitin sulfate as a contaminant in heparin preparations.
复制标题
DOI：
10.1016/j.ab.2009.02.012
复制
发表时间：
2009-05-01
影响因子：
2.9
通讯作者：
Linhardt RJ
中科院分区：
文献类型：
--
作者：
Volpi N;Maccari F;Linhardt RJ
本文描述了一种简单、准确、可靠的定量测定肝素制剂中过硫酸软骨素（OSCS）的方法。多糖经酸性水解降解后，产生的己糖胺，即Hep的GlcN和OSCS的GalN，与邻氨基苯甲酸（AA）衍生，并通过CE分离。10 min，在214 nm高灵敏度检测(检出限（LOD）约为。200 pg)。此外，aa衍生的GlcN和GalN表现出相当相似的摩尔吸收率，允许直接和简单地定量Hep样品中的OSCS。此外，由于Hep样品中可能存在天然硫酸软骨素和硫酸皮聚糖杂质，因此可以采用软骨素ABC裂解酶进行特异性酶处理的初步步骤，以特异性消除分析中的干扰，使该分析方法对OSCS污染具有高度特异性，因为软骨素ABC裂解酶无法作用于这种半合成聚合物。对该方法的特异性、线性度、准确度、精密度、检出限和定量限进行了验证。由于CE具有很高的灵敏度，Hep样品中可以检测到1%的OSCS污染物。最后，一份被污染的Hep生样品被发现含有38.9%的OSCS，而一份配制的Hep被计算出含有39.7%的OSCS。
A simple, accurate and robust quantitative CE method for the determination of oversulfated chondroitin sulfate (OSCS) as a contaminant in heparin (Hep) preparations is described. After degradation of the polysaccharides by acidic hydrolysis, the hexosamines produced, i.e., GlcN from Hep and GalN from OSCS, were derivatized with anthranilic acid (AA) and separated by means of CE in approx. 10 min with high sensitivity detection at 214 nm (limit of detection (LOD) of approx. 200 pg). Furthermore, AA-derivatized GlcN and GalN showed quite similar molar absorptivity allowing for direct and simple quantification of OSCS in Hep samples. Moreover, a preliminary step of specific enzymatic treatment by using chondroitin ABC lyase may be applied for the specific elimination of interference in the analysis due to the possible presence in Hep samples of natural chondroitin sulfate and dermatan sulfate impurities, making this analytical approach highly specific for OSCS contamination, since chondroitin ABC lyase is unable to act on this semi-synthetic polymer. The CE method was validated for specificity, linearity, accuracy, precision, LOD and limit of quantification (LOQ). Due to the very high sensitivity of CE, as little as 1% OSCS contaminant in Hep sample could be detected and quantified. Finally, a contaminated raw Hep sample was found to contain 38.9% OSCS while a formulated contaminated Hep was calculated to have 39.7% OSCS.
登录
查看更多内容
影响因子：
46.9
作者：
Guerrini, Marco;Beccati, Daniela;Sasisekharan, Ram
通讯作者：
Sasisekharan, Ram
影响因子：
2.9
作者：
Volpi, Nicola;Maccari, Francesca;Linhardt, Robert J.
通讯作者：
Linhardt, Robert J.
影响因子：
3
作者：
Lindahl, U
通讯作者：
Lindahl, U
影响因子：
3.8
作者：
NEVILLE, GA;MORI, F;PERLIN, AS
通讯作者：
PERLIN, AS
影响因子：
1.6
作者：
Sato, K;Sato, K;Yamazaki, S
通讯作者：
Yamazaki, S