基于PMP-HPLC和化学计量学的黄精基原物种多糖差异分析

目的:建立2015年版《中国药典》中3种基原黄精的多糖指纹图谱,探讨黄精基原物种多糖的差异,为黄精药材质量评价和临床应用提供参考。方法:采用水提醇沉法提取黄精药材中的多糖,经三氟乙酸(TFA)水解和1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生后,采用HPLC建立3种黄精的色谱指纹图谱;并利用相似度(SA)分析,聚类分析(HCA)和主成分分析方法(PCA)对指纹图谱进行分析,研究3种基原黄精物种多糖的差异。结果:3种基原黄精的多糖PMP-HPLC指纹图谱存在差异,3个物种中均未检测到D-甘露糖,L-鼠李糖和L-岩藻糖,但均含有D-半乳糖醛酸,D-盐酸氨基葡萄糖,D-半乳糖,D-葡萄糖,D-木糖。PCA和HCA分析表明,黄精与多花黄精的多糖色谱指纹较为接近,而滇黄精与二者差异明显。结论:3种法定基原黄精物种的多糖组成存在差异,在临床应用中,应考虑物种对临床应用可能产生的影响。所建立PMP-HPLC方法简便、准确、重复性好,可用于3种黄精药材的多糖的差异研究。

Approach Based on PMP-HPLC Fingerprint and Chemometrics to Compare Difference of Polysaccharide in Polygonati Rhizoma

Objective:To established an approach of chemical fingerprinting and study the differences of the polysaccharides from three species of Polygonati Rhizoma in Chinese Pharmacopoeia,so as to provide reference for quality evaluation and clinical application of Polygonati Rhizoma. Method:The polysaccharides were extracted by water extraction and alcohol precipitation from Polygonati Rhizoma. After hydrolysis by trifluoroacetic acid (TFA) and pre-column derivation by PMP,the chromatographic fingerprints of three kinds of Polygonati Rhizoma were established by high performance liquid chromatography. The fingerprinting model and chemometrics method,include similarity analysis (SA),cluster analysis (HCA) and principal component analysis (PCA) were used for compare the differences among three species. Result:There were some differences in the PMP-HPLC fingerprints and monosaccharide composition from the three species. The D-mannose,L-rhamnose and L-fucose were not detected,but they all contained D-galacturonic acid,D-glucosamine hydrochloride,D-galactose,D-glucose and D-xylose among three species. The PCA and HCA analysis showed that chromatographic fingerprints of P. cyrtonema and P. sibiricum were similar,while P. kingianum and other two species were significantly different. Conclusion:There are differences in fingerprints of polysaccharides among three species of Polygonati Rhizoma. The possible effects of species should be considered in clinical application. The established PMP-HPLC is a simple,accurate and reproducible method,which can be used for the quality evaluation of Polygonati Rhizoma.