不同产地滇黄精高效液相色谱法指纹图谱的建立及其多糖含量测定Establishment of high performance liquid chromatography fingerprint and determination of polysaccharide content in Polygonatum kingianum from different origins
马琴,郭思潍,杜强,杨国海,王才武,周雪,吴林菁
MA Qin,GUO Siwei,DU Qiang,YANG Guohai,WANG Caiwu,ZHOU Xue,WU Linjing
摘要(Abstract):
目的 探讨不同产地滇黄精高效液相色谱法(HPLC)指纹图谱的建立及其多糖含量的测定。方法 取贵州(剑河、安顺、天柱、铜仁、赤水、贵阳及纳雍)、广西百色及四川内江来源的9个批次滇黄精药材各1 g制备供试品溶液,采用DiKMA Platisll ODS C_(18)色谱柱(流动相为乙腈-0.05%磷酸水溶液、梯度洗脱,流速1 mL/min,柱温35℃,进样量20μL,检测波长203 nm)建立滇黄精HPLC指纹图谱,并采用相似度评价、主成分分析及聚类分析等方法评价不同产地滇黄精的质量;采用蒽酮-硫酸法测定各批次滇黄精的多糖含量。结果 建立了9批滇黄精的HPLC指纹图谱,共确认12个共有峰;9批不同产地的滇黄精多糖含量均大于7%,含量顺序为贵州剑河>贵州安顺>贵州天柱>贵州铜仁>贵州赤水>贵州贵阳>贵州纳雍>广西百色>四川内江;相似度结果显示,四川内江、贵州贵阳及广西百色的相似度分别为0.762、0.809及0.846,贵州其余地区滇黄精的相似度≥0.853;主成分分析结果显示共得到3个主成分因子,聚类分析结果表明滇黄精被分为3类,且滇黄精多糖含量≥7.16%。结论 本研究建立的滇黄精HPLC指纹图谱及黄精多糖成分含量的测定方法简便可行,准确可靠。
Objective To investigate establishment of high performance liquid chromatography(HPLC) fingerprints and determination of polysaccharides content in Polygonatum kingianum from different origins. Methods Nine batches of Polygonatum kingianum(1 g) from Guizhou(Jianhe, Anshun, Tianzhu, Tongren, Chishui, Guiyang, and Nayong), Baise Guangxi, and Neijiang Sichuan sources were taken to prepare the test solution(1 g from each origin). The separation was performed on a DiKMA Platisll ODS C_(18)(250 mm×4.6 mm, 5 μm) column(with acetonitrile-0.05% phosphoric acid aqueous solution as mobile phase in gradient elution at a flow rate of 1 mL/min and a column temperature of 35 ℃ with an injection volume of 20 μL and a detection wavelength of 203 nm) to establish HPLC fingerprints of Polygonatum kingianum extracts; the quality of Polygonatum kingianum from different origins was evaluated by similarity evaluation, principal component analysis and cluster analysis; the content of polysaccharides of Polygonatum kingianum from each batch was determined by anthrone-sulfuric acid method. Results HPLC fingerprints were established for nine batches of Polygonatum kingianum, and a total of 12 common peaks were identified. The polysaccharide content in all nine batches from different sources exceeded 7%. Among them, the descending order of polysaccharide content was as follows: Jianhe Guizhou>Anshun Guizhou>Tianzhu Guizhou>Tongren Guizhou>Chishui Guizhou>Nayong Guizhou>Baise Guangxi>Neijiang Sichuan. The similarity analysis results indicated that the similarities between Neijiang Sichuan, Guiyang Guizhou and Baise Guangxi were 0.762, 0.809, and 0.846, respectively; while the similarity of Polygonatum kingianum between other areas in Guizhou province were ≥0.853. Furthermore, three principal component factors were obtained using principal component analysis. Cluster analysis revealed that Polygonatum kingianum could be categorized into three distinct groups and content of polysaccharides of Polygonatum kingianum were ≥7.16%. Conclusion The method of HPLC fingerprints and determination of polysaccharides content in Polygonatum kingianum is simple, feasible, accurate, and reliable.
关键词(KeyWords):
聚类分析;主成分分析;滇黄精;指纹图谱;高效液相色谱法;紫外分光光度法
cluster analysis;principal component analysis;Polygonatum kingianum;fingerprint;high performance liquid chromatography;ultraviolet spectrophotometry
基金项目(Foundation): 贵州省科技计划项目(黔科合基础-ZK[2022]一般389,黔科合支撑[2020]4Y104,黔科合支撑[2023]一般046);; 贵州省科技创新基地(黔科合中引地[2023]003);; 贵州省高层次创新型人才百层次人才(黔科合平台人才-GCC[2023]048)
作者(Author):
马琴,郭思潍,杜强,杨国海,王才武,周雪,吴林菁
MA Qin,GUO Siwei,DU Qiang,YANG Guohai,WANG Caiwu,ZHOU Xue,WU Linjing
DOI: 10.19367/j.cnki.2096-8388.2024.03.009
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