一测多评法测定大鼠肠温孵液中意大利牛舌草4种成分的含量

摘要

Objective To establish a quantitative analysis of multi-components by single marker (QAMS) for the determination of four components (rutin,kaempferol-3-O-rutnoside,narcissin and rosmarinic acid) in Anchusa italica Retz.in rat intestinal incubation solution.Methods The compounds from the Anchusa italica Retz.crude extracts were purified by the polyamide column chromatography.An HPLC method was developed for the determination of rutin,kaempferol-3-O-rutnoside,narcissin and rosmarinic acid of Anchusa italica Retz.in rat intestinal incubation solution in simultaneously.Rutin was selected as the internal reference substance.Relative correction factors (RCF) of kaempferol-3-O-rutnoside,narcissin,and rosmarinic acid to rutin were established.Obtained RCFs were used to calculate content (calculated value).At the same time,the contents (measured value) of the three components were also determined by external standard method (ESM).The quantitative results were compared between the ESM and QAMS,the feasibility of QAMS method were verified.Results The average contents of the 4 main components in rat intestinal incubation solution of 8 batchs were 14.20 mg/g,15.88 mg/g,9.80 mg/g and 66.90 mg/g respectively.Within the certain range,the RCFs of kaempferol-3-O-rutnoside,narcissin,and rosmarinic acid to rutin were 0.983,1.101 and 0.843 (the values of RSD＜2％).The mean calculated value of kaempferol-3-O-rutnoside,narcissin,rosmarinic acid were 15.67 mg/g,9.87 mg/g and 67.44 mg/g respectively.The results of the determination of the two methods are basically the same (the relative deviation＜2 ％).Conclusion This method can be used to the determination of kaempferol-3-O-rutnoside,narcissin,and rosmarinic acid in Anchusa italica Retz.in rat intestinal incubation solution.%目的 建立大鼠肠温孵液中意大利牛舌草4种成分(芦丁、山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸)的一测多评方法(Quantitative Analysis of Multi-components by Single-marker,QAMS).方法 采用聚酰胺柱色谱对意大利牛舌草粗提物进行纯化.建立同时测定大鼠肠温孵液中意大利牛舌草提取物主要成分芦丁、山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸的高效液相色谱定量分析方法.以芦丁为内参物,测定大鼠肠温孵液中意大利牛舌草提取物主要成分芦丁与山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸的相对校正因子(Relative Correction Factor,RCF),利用RCF进行含量计算(计算值),并与外标法(External Standard Method,ESM)的测定结果(实测值)进行比较,对该方法进行可行性验证.结果 采用ESM法测定8个批次的大鼠肠温孵液中意大利牛舌草提取物主要成分芦丁、山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸的平均含量分别为14.20、15.88、9.80、66.90 mg/g.在线性范围内,芦丁与山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸的RCF分别为0.983、1.101、0.843(RSD值均＜2％).利用RCF计算得山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸的平均含量分别为15.67、9.87、67.44 mg/g,两种方法的含量测定结果基本一致(相对偏差均＜2％).结论 一测多评法简便、准确、可行,可用于大鼠肠温孵液中意大利牛舌草提取物山奈酚-3-O-芸香糖苷、水仙苷、迷迭香酸的含量测定.