建立并应用一种快速高效的 CZE 方法，并结合校正因子，用于测定来自麻黄的酸性杂多糖的单糖组成。
Development and application of a rapid and efficient CZE method coupled with correction factors for determination of monosaccharide composition of acidic hetero-polysaccharides from Ephedra sinica.
机构信息
Pharmaceutical College, Heilongjiang University of Chinese Medicine, 24 Heping Road, Harbin 150040, People's Republic of China.
出版信息
Phytochem Anal. 2011 Mar-Apr;22(2):103-11. doi: 10.1002/pca.1235. Epub 2010 Dec 30.
INTRODUCTION
Ephedrine alkaloids cannot account for all the effects of Ephedra sinica and the polysaccharides are also demonstrated to be one of the main bioactive constituents of E. sinica. However, no work has been reported on the analysis of monosaccharide composition of purified polysaccharides isolated from the stem of E. sinica.
OBJECTIVE
To develop a rapid and efficient capillary zone electrophoresis (CZE) method based on pre-column derivatisation with 1-phenyl-3-methyl-5-pyrazolone for the simultaneous determination of neutral and acidic sugars of purified polysaccharides from E. sinica.
METHODOLOGY
Three polysaccharides (ESP-A3, ESP-A4 and ESP-B4) were isolated and purified by ion exchange and gel-filtration chromatography from the stem of E. sinica. The effects of background electrolyte pH and concentration, applied voltage and temperature on the separation were investigated. Meanwhile, factors affecting the hydrolysis of ESP-B4 with sulphuric acid were investigated by changing the hydrolysis time, acid concentration and hydrolytic temperature to achieve complete hydrolysis. The standard curves coupled with correction factors were used to calculate molar ratios.
RESULTS
The optimal CZE method coupled with correction factors was successfully applied to the determination of molar ratios of three purified polysaccharides and their corresponding partial acid hydrolysis products. ESP-A3, ESP-A4 and ESP-B4 were all typical acidic hetero-polysaccharides and consisted of xylose, arabinose, glucose, rhamnose, mannose, galactose, glucuronic acid and galacturonic acid, and their corresponding molar ratios were 6.8:7.5:1.0:14.0:13.7:22.3:10.2:3.8 for ESP-A3, 1.2:4.1:1.0:5.1:1.6:17.3:3.1:2.2 for ESP-A4, and 1.0:4.5:1.0:2.0:1.0:5.5:1.5:50.0 for ESP-B4.
CONCLUSION
The results provided scientific evidence for the further study of the structure and bioactivity of complex acidic E. sinica polysaccharides.
简介
麻黄生物碱不能解释麻黄的所有作用，多糖也被证明是麻黄的主要生物活性成分之一。然而，目前还没有关于从麻黄茎中分离得到的纯化多糖的单糖组成分析的报道。
目的
建立一种快速、高效的毛细管区带电泳（CZE）方法，通过 1-苯基-3-甲基-5-吡唑啉酮柱前衍生，同时测定从麻黄茎中分离得到的纯化多糖的中性糖和酸性糖。
方法
采用离子交换和凝胶过滤色谱法从麻黄茎中分离纯化了三种多糖（ESP-A3、ESP-A4 和 ESP-B4）。考察了背景电解质 pH 值和浓度、外加电压和温度对分离的影响。同时，通过改变水解时间、酸浓度和水解温度，考察了硫酸水解 ESP-B4 的影响因素，以达到完全水解。采用标准曲线和校正因子计算摩尔比。
结果
成功地将优化的 CZE 方法与校正因子相结合，应用于三种纯化多糖及其部分酸水解产物摩尔比的测定。ESP-A3、ESP-A4 和 ESP-B4 均为典型的酸性杂多糖，由木糖、阿拉伯糖、葡萄糖、鼠李糖、甘露糖、半乳糖、葡萄糖醛酸和半乳糖醛酸组成，其摩尔比分别为 6.8:7.5:1.0:14.0:13.7:22.3:10.2:3.8 对于 ESP-A3、1.2:4.1:1.0:5.1:1.6:17.3:3.1:2.2 对于 ESP-A4，以及 1.0:4.5:1.0:2.0:1.0:5.5:1.5:50.0 对于 ESP-B4。
结论
该结果为进一步研究复杂酸性麻黄多糖的结构和生物活性提供了科学依据。
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