多糖部分降解产物在板蓝根颗粒多糖成分分析中的应用

Application of partial degradation products of polysaccharide in analysis of polysaccharide from Banlangen Granules

板蓝根多糖的药理活性与板蓝根的功效一致,其部分降解产物的毛细管区带电泳(CZE)图谱特征性强,可用于板蓝根颗粒中多糖类成分的定性分析.首先采用酶法去除板蓝根颗粒中糊精等辅料的干扰,所得板蓝根多糖在盐酸浓度0.5 mol/L,温度80℃条件下部分降解4 h,以1-苯基-3-甲基-5-吡唑啉酮(PMP)为衍生试剂,采用毛细管区带电泳法测定其部分降解产物图谱.电泳条件:未涂层弹性石英毛细管柱(70 cm×50μm,i.d,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245 nm,运行电压17 kV,室温下重力进样10 cm×15 s,进而运用聚类分析(CA)对该产物的CZE谱图进一步探讨.结果表明:CZE图谱在低聚糖部分具有差异,而单糖组成相对稳定,均含木糖、阿拉伯糖、葡萄糖、鼠李糖、甘露糖和半乳糖等,结合板蓝根药材相应谱图,从低聚糖和单糖上进行比对可达到鉴别目的,且发现不同厂家产品之间多糖含量存在较大差异.板蓝根颗粒中多糖部分降解产物CZE图谱的建立完善了该药品的质量控制,同时也为其药效物质研究及其他相关中药制剂中多糖的质量控制提供参考.

The pharmacological activities of Radix isatidis polysaccharide are consistent with the efficacy of R.isatidis.As the capillary zone electrophoresis(CZE)spectrum of partial degradation products of R.isatidis polysaccharide has obvious characteristics,and can be used for the qualitative analysis of polysaccharide from Banlangen Granules.In this paper,the interference of pharmaceutical excipients such as dextrin from Banlangen Granules was firstly removed by enzymatic method.Then the obtained R.isatidis polysaccharide was partially degraded for 4 h at a hydrochloric acid concentration of 0.5 mol/L and a temperature of 80℃,and the spectra were determined by CZE 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×50μm i.d.,61 cm effective length),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 17 kV,hydrodynamic pressure injection was employed(10 cm×15 s).The CZE spectra of partial degradation products were further explored using cluster analysis(CA).The results showed that the CZE spectra had some differences in oligosaccharide composition,while monosaccharide composition was relatively stable,including xylose,arabinose,glucose,rhamnose,mannose and galactose.Combined with the corresponding spectrum of R.isatidis,we could achieve identification purpose through comparison between their oligosaccharides and monosaccharides,and there were significant differences in polysaccharide contents among the products of different manufacturers.The establishment of CZE spectrum of the partial degradation products of polysaccharide from Banlangen Granules has improved the quality control of the drug,and also provided reference for the study of the pharmacodynamic substances and the quality control of polysaccharide in other related Chinese medicine preparations.