摘要
目的采用毛细管区带电泳法(CZE)测定不同虫草菌粉制剂中多糖的单糖组成。方法设计正交试验优化制剂中所提取多糖的全降解条件;以1-苯基-3-甲基-5-吡唑啉酮(PMP)为衍生试剂,采用CZE法测定其单糖组成;电泳条件:未涂层弹性石英毛细管柱(70 cm×50μm,有效长度61 cm);运行缓冲液为40 mmol/L硼砂缓冲液(pH 10.1);检测波长245nm;运行电压13 kV;重力进样(10 cm×8 s)。结果多糖全降解的最优条件为硫酸浓度1.0 mol/L,温度100℃,时间5 h;心肝宝胶囊、百令胶囊、至灵菌丝胶囊和金水宝胶囊的多糖中均含有木糖、葡萄糖、甘露糖和半乳糖4种单糖,但含量各有差异;4种制剂中多糖的单糖组成与冬虫夏草多糖中的单糖组成基本一致,且检测出了冬虫夏草中未见报道的木糖、鼠李糖和葡萄糖醛酸等单糖。对所建立的CZE法进行方法学验证,结果显示各单糖在0.01～0.20mg/m L线性关系良好,检测限0.205～0.397μg/m L,定量限0.684～1.323μg/m L,精密度RSD为0.8%～3.6%,重复性RSD为2.7%～4.7%,稳定性RSD为2.8%～4.7%。葡萄糖和半乳糖的平均回收率分别为96.1%～99.8%和95.1%～99.6%,其RSD分别为1.0%～2.0%和1.5%～1.9%。结论该方法快速高效,为虫草菌粉制剂质量标准的完善和虫草菌粉替代冬虫夏草的深入研究提供参考。
Objective To determine the monosaccharide composition of polysaccharides in different Cordyceps powder preparations by capillary zone electrophoresis(CZE). Methods The orthogonal experiment was used to optimize the total degradation conditions of the extracted polysaccharides;The monosaccharide composition was determined by CZE method using 1-phenyl-3-methyl-5-pyrazolone(PMP) as a derivatizing reagent. The electrophoresis conditions were as follows: uncoated fused silica capillary column(70 cm × 50 μm i.d., effective length was 61 cm);40 mmol/L borax solution(pH 10.1) was used as the running buffer;The detection wavelength was set at 245 nm;The operation voltage was 13 kV;Hydrodynamic pressure injection was employed(10 cm × 8 s). Results The optimum hydrolysis conditions were as follows: 1.0 mol/L sulfuric acid solution, hydrolysis temperature 100 ℃, and hydrolysis time 5 h. The common monosaccharides of polysaccharides in Xinganbao Capsule, Bailing Capsule, Zhiling Junsi Capsule, and Jinshuibao Capsule were xylose, glucose, mannose, and galactose, but the content was different. The monosaccharide components contained in the polysaccharides of the four preparations were basically the same as those of the Cordyceps sinensis, and monosaccharides included xylose, rhamnose and glucuronic acid which were not reported in Cordyceps sinensis were detected. Each monosaccharide had a good linear relationship with the concentration range of 0.01 to 0.20 mg/mL, the limits of detection(LOD, S/N = 3) and the limits of quantification(LOQ, S/N = 10) ranged from 0.205 to 0.397 μg/m L and 0.684 to 1.323 μg/mL, respectively. RSDs of the precision test were 0.8%-3.6%, RSDs of the repeatability test were 2.7%-4.7%, and 2.8%-4.7% in the stability test. The recovery rate of the method showed that the mean recoveries of glucose and galactose ranged from 96.1% to 99.8% and 95.1% to 99.6% respectively, and RSD values fell within 1.0%-2.0% and 1.5%-1.9%, respectively. Conclusion The method is fast and efficient, and provides a refer
作者
张建
卢凯
刘艳萍
荣雨晨
郭怀忠
ZHANG Jian;LU Kai;LIU Yan-ping;RONG Yu-chen;GUO Huai-zhong(College of Pharmacy,Hebei University,Baoding 071002,China;Key Laboratory of Pharmaceutical Quality Control of Hebei Province,Baoding 071002,China;Key Laboratory of Medicinal Chemistry and Molecular Diagnosis,Ministry of Education,Baoding 071002,China)
出处
《中草药》
CAS
CSCD
北大核心
2019年第1期90-96,共7页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(21575033)
河北省自然科学基金资助项目(H2016201221)
河北省自然科学基金资助项目(B2018201270)
河北省高等学校科学技术研究项目(ZD2015036)
河北大学精品实验项目(2017-BZ-JPSY25)
关键词
虫草菌粉制剂
虫草多糖
单糖组成
毛细管区带电泳法
质量控制
Cordyceps powder preparation
Cordyceps polysaccharides
monosaccharide composition
capillary zone electrophoresis
quality control
