基于PMP-HPLC和化学计量学的黄精基原物种多糖差异分析
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摘要
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。
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.
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.
引文
[1]国家药典委员会.中华人民共和国药典.一部[M].北京:中国医药科技出版社,2015:306.
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[4]王坤,岳永德,汤锋,等.多花黄精多糖的分级提取及结构初步分析[J].天然产物研究与开发,2014,26(3):364-369.
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[7]余亚鸣,马晓勇,张铁军,等.基于高效液相色谱-质谱联用技术的黄精化学成分快速鉴别研究[J].时珍国医国药,2016,27(4):794-797.
[8]杨兴鑫,董金财,胡海波,等.基于多元统计分析的3种法定基源黄精化学成分比较研究[J].中国中医药信息杂志,2018,25(5):71-76.
[9]姜程曦,张铁军,陈常青,等.黄精的研究进展及其质量标志物的预测分析[J].中草药,2017,48(1):1-16.
[10]邝婷婷,王宇,王张,等.柱前衍生HPLC法分析蔓菁多糖中单糖的组成[J].中成药,2014,36(10):2121-2124.
[11]柳杨,李清,王菲,等.柱前衍生HPLC法分析藤三七多糖中单糖的组成[J].中药新药与临床药理,2011,22(2):199-202.
[12]郝燕,董鸿晔,姜楠,等.基于主成分分析的中药色谱指纹图谱多维多息特征数据挖掘方法研究[J].中南药学,2007,5(3):267-272.
[13]郭龙,焦倩,张丹,等.基于指纹图谱和多组分含量测定的艾叶药材质量控制研究[J].中国中药杂志,2018,43(5):977-984.
[14]张晓灿,罗丹丹,陶爱恩,等.基于指纹图谱和化学计量学的丹参及紫丹参质量评价研究[J].中药材,2017,40(5):1061-1065.
[2]陶爱恩,张晓灿,杜泽飞,等.黄精属植物中黄酮类化合物及其药理活性研究进展[J].中草药,2018,49(9):2163-2171.
[3]苏文田,刘跃钧,蒋燕锋,等.黄精产业发展现状与可持续发展的建议[J].中国中药杂志,2018,43(13):2831-2835.
[4]王坤,岳永德,汤锋,等.多花黄精多糖的分级提取及结构初步分析[J].天然产物研究与开发,2014,26(3):364-369.
[5]秦垂新,曹子丰,黄绮敏,等.黄精多糖水解物柱前衍生HPLC指纹图谱[J].中国实验方剂学杂志,2015,21(11):65-68.
[6]王海洋,高阳,高其品,等.黄精多糖的柱前衍生化HPLC指纹图谱研究[J].湖北农业科学,2016,55(13):3462-3464,3478.
[7]余亚鸣,马晓勇,张铁军,等.基于高效液相色谱-质谱联用技术的黄精化学成分快速鉴别研究[J].时珍国医国药,2016,27(4):794-797.
[8]杨兴鑫,董金财,胡海波,等.基于多元统计分析的3种法定基源黄精化学成分比较研究[J].中国中医药信息杂志,2018,25(5):71-76.
[9]姜程曦,张铁军,陈常青,等.黄精的研究进展及其质量标志物的预测分析[J].中草药,2017,48(1):1-16.
[10]邝婷婷,王宇,王张,等.柱前衍生HPLC法分析蔓菁多糖中单糖的组成[J].中成药,2014,36(10):2121-2124.
[11]柳杨,李清,王菲,等.柱前衍生HPLC法分析藤三七多糖中单糖的组成[J].中药新药与临床药理,2011,22(2):199-202.
[12]郝燕,董鸿晔,姜楠,等.基于主成分分析的中药色谱指纹图谱多维多息特征数据挖掘方法研究[J].中南药学,2007,5(3):267-272.
[13]郭龙,焦倩,张丹,等.基于指纹图谱和多组分含量测定的艾叶药材质量控制研究[J].中国中药杂志,2018,43(5):977-984.
[14]张晓灿,罗丹丹,陶爱恩,等.基于指纹图谱和化学计量学的丹参及紫丹参质量评价研究[J].中药材,2017,40(5):1061-1065.