不同产地丫蕊花不同部位中3种皂苷成分含量测定及差异分析
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篇名: 不同产地丫蕊花不同部位中3种皂苷成分含量测定及差异分析
TITLE: Content Determination and Difference Analysis of 3 Saponins in Different Parts of Ypsilandra thibetica from Different Producing Areas
摘要: 目的:建立测定丫蕊花中重楼皂苷Ⅱ、Ⅵ、Ⅶ含量的方法,比较不同产地丫蕊花不同部位中这3种皂苷成分的含量差异。方法:采用高效液相色谱法测定10个产地丫蕊花的全草部位和地下部位中重楼皂苷Ⅱ、Ⅵ、Ⅶ的含量,色谱柱为KromasilC18,流动相为乙腈-水(梯度洗脱),流速为1.0mL/min,柱温为35℃,检测波长为203nm,进样量为10μL。以3种皂苷成分的含量为指标,对20批次丫蕊花进行聚类分析和偏最小二乘判别分析,对样品进行聚类区分,并确定其主要差异性成分。结果:重楼皂苷Ⅱ、重楼皂苷Ⅵ、重楼皂苷Ⅶ的检测质量线性范围分别为0.051~2.04、0.007~0.28、0.168~6.72μg(r≥0.9995);检测限分别为1.92、1.75、1.87ng,定量限分别为6.40、5.87、6.23ng;精密度、重复性、稳定性(24h)的RSD均小于2%(n=6);平均回收率分别为99.29%、101.38%、99.64%,RSD分别为1.17%、2.64%、0.75%(n=6)。20批次样品的重楼皂苷Ⅱ含量为0.615~1.875mg/g,重楼皂苷Ⅵ含量为0~0.095mg/g,重楼皂苷Ⅶ含量为3.158~12.354mg/g。聚类分析结果显示,20批次样品被聚为两类,批次S9~S12为一类,其余16批次样品被聚为一类;偏最小二乘判别分析结果显示,20批次样品分为3个区,批次S1、S2、S8、S14、S16、S20被分为Ⅰ区,批次S9~S12被分为Ⅱ区,其余样品被分为Ⅲ区。不同产地的丫蕊花存在品质差异,且全草和地下部位中3种皂苷成分无差异性。权重排序结果显示,重楼皂苷Ⅶ为丫蕊花药材中主要差异性成分,其中彭州和大邑县产丫蕊花药材中重楼皂苷Ⅶ最高。结论:本方法简单方便、灵敏度高,可用于丫蕊花中3种皂苷成分的含量测定。以彭州和大邑县产丫蕊花活性成分含量较高、质量较好。
ABSTRACT: OBJECTIVE:To establish the method for content dete rmination of polyphyllin Ⅱ,Ⅵ,Ⅶ in Ypsilandra thibetica , and to compare the differences of 3 saponins in different parts of Y. thibetica from different producing areas. METHODS :HPLC method was adopted to determine the contents of polyphyllin Ⅱ,Ⅵ,Ⅶ in whole grass part and underground part of Y. thibetica from 10 producing areas. The determination was performed on Kromasil C 18column with mobile phase consisted of acetonirile-water (gradient elution )at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and detection wavelength was set at 203 nm;sample size was 10 μL. With the contents of 3 saponins as the index ,20 batches of Y. thibetica were analyzed by cluster analysis and PLS-DA analysis ;the aggregation of samples was analyzed and determined the primary difference components. RESULTS:The linear range of polyphyllin Ⅱ,polyphyllin Ⅵ and polyphyllin Ⅶ were 0.051-2.04,0.007-0.28,0.168-6.72 μg, respectively(r≥0.999 5);the detection limits were 1.92,1.75,1.87 ng,respectively;and the quantitative limits were 6.40,5.87, 6.23 ng,respectively;RSD of precision ,reproducibility and stability tests (24 h)were all lower than 2%(n=6);the average recovery rates were 99.29%,101.38% and 99.64%,with RSDs of 1.17%,2.64%,0.75%(n=6),respectively. The content of polyphyllin Ⅱ was 0.615-1.875 mg/g,that of polyphyllin Ⅵ was 0-0.095 mg/g,and that of polyphyllin Ⅶ was 3.158-12.354 mg/g. Cluster analysis showed that 20 batches of samples were clustered into two groups ,batch S 9-S12 were clustered in to one group,and the other 16 batches of samples were clustered into another group. PLS-DA analysis showed that 20 batches of samples were divided into 3 areas,batch S 1,S2,S8,S14,S16,S20 were included in area Ⅰ;batch S 9-S12 included in area Ⅱ;and the rest of the samples included in area Ⅲ. The quality of Y. thibetica from different habitats was different ,and there was no difference in the saponin composition between the whole grass and the underground part. Weight ranking found that mail:cscdtcm@126.com polyphyllin Ⅶ was the main difference component in Y. thibetica,and the content of polyphyllin Ⅶ in Y. thibetica from Pengzhou city and Dayi county was the highest. CONCLUSIONS :The established method is simple ,convenient and sensitive. It can be used for the content determination of 3 saponins in Y. thibetica . The content of active components is higher and the quality is better in Y. thibetica from Pengzhou city and Dayi county.
期刊: 2020年第31卷第03期
作者: 陈帅,张美,袁崇均,罗森,王笳
AUTHORS: CHEN Shuai,ZHANG Mei,YUAN Chongjun ,LUO Sen,WANG Jia
关键字: 高效液相色谱法;丫蕊花;重楼皂苷Ⅱ;重楼皂苷Ⅵ;重楼皂苷Ⅶ;聚类分析;偏最小二乘判别分析
KEYWORDS: HPLC;Ypsilandra thibetica ;Polyphyllin Ⅱ;Polyphyllin Ⅵ;Polyphyllin Ⅶ;Cluster analysis ;PLS-DA analysis
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