HPLC同时测定墨旱莲正丁醇有效部位含量分析

Simultaneous Determination of the Active Contents in N-butanol Effective Fraction of Herba Ecliptae by HPLC

目的建立HPLC法定量同时分析中药材墨旱莲有效部位成分的方法,为墨旱莲的质量控制提供参考。方法根据临床文献对墨旱莲正丁醇提取部位活性成分药理筛选实验结果,确定墨旱莲正丁醇有效部位主要活性成分为(墨旱莲皂甙A、齐墩果酸、芹菜素、木犀草素、α-三联噻吩、(丁烯-3-炔-卜基)2,2-二联噻吩、蟛蜞菊内酯、去甲基蟛蜞菊内酯)。选择市售经鉴定的墨旱莲,采用经石油醚-乙酸乙酯-正丁醇提取步骤得到墨旱莲正丁醇有效部位,采用高效液相色谱法(HPLC)对其主要活性成分进行分离、同时检测。流动相组成:洗脱程序:。采用8种待测成分的对照品,对该HPLC方法检测8种成分的线性关系、加样回收率变异系数、日间精密度变异系数、重复率变异系数、稳定性变异系数进行分析,综合评价HPLC法定量分析墨旱莲有效部位8种成分的价值。结果采用上述HPLC条件检测墨旱莲正丁醇有效部位的8种活性成分,结果显示,8种成分墨旱莲皂甙A、齐墩果酸、芹菜素、木犀草素、α-三联噻吩、(丁烯-3-炔-卜基)2,2-二联噻吩、蟛蜞菊内酯、去甲基蟛蜞菊内酯的峰面积和浓度均呈线性关系,相关性系数均>0.999,8种成分的加样回收率均>98.0%,8种成分的日间精密度变异系数、稳定性、重复率变异系数均<3.0%。结论 HPLC法同时测定墨旱莲有效部位的8种活性成分具有较好的稳定性,可重复性好,回收率高。为墨旱莲有效部位活性成分的质量控制提供了量化手段,是适合墨旱莲临床主要以有效部位作为临床应用的特性,具有较高的应用价值。

Objective To establish an HPLC method for the simultaneous quantitation of the active contents in the n-butanol effective fraction of Herba Eelipta components would provide references for the qualiiy control. Methods According to the clinical researches about the pharmacological screening of the active contents in n-butanol effective fraction of Herbe Ecliptae,determined the main active components（including ginsenoside A, oleanolic acid, apigenin, ]uteolin, alpha terthienyl, （3 butene alkyne group 2,2 BU） one or two bithiophene,W, parthenolide and demethylwedelolactone）. Commercially available and identified Eclipta was extracted with petroleum ether, ethyl acetate and normal butanol successively to get n-butanol effective fraction, the main active components in which were separated and detected by high performance liquid chromatography （HPLC） simultaneously. Mobile phase composition ：elution procedure. Using the standard solutions of the eight components, the linearity, recovery,inter day precision, reproducibility and stability of the HPLC method for simultaneous determination of the 8 components were analyzed, and the value of the method was synthetically assessed. Results Using the above detection condition, the results showed that the relationship be- tween peak area and concentration of eight active components,including ginsenoside A,oleanolic acid ,apigenin,luteolin, alpha terthienyl, （3 butene alkyne Bu Ji） 2, peak area and concentration of 2 one or two bithiophene,wedelolactone and demethylwedelo- lactone were linear dependence, and the correlation coefficients were all greater than 0.999. The recoveries of the eight components were a]] over 98% ,and the variation coefficient of the precision,stability and repetition rate were all less than 3%. Conclusion The HPLC method for the simultaneous determination of the eight active components in the n-butanol effective fraction of Eclipta had well stability and repeatability and high recovery rate. We provided and validated a quantitative method for the quality control of the active ingredients in the effective fraction of Eclipta, which was suitable for clinical characteristics of applying clinical main effective contents in Eclipta and had higher application value.