高效液相色谱法测定不同采收期余甘子叶中5种酚酸类成分含量变化

Changes of Contents of Five Phenolic Acids Ingredients in Phyllanthus emblica L.during Different Harvesting Times by HPLC

目的:建立同时测定不同采收期余甘子叶药材中5种酚酸类成分含量的方法,研究其变化规律,确定最佳采收期。方法:利用高效液相色谱法测定不同采收期余甘子叶药材中表没食子儿茶素、柯里拉京、诃子联苯酸、鞣花酸和诃子酸的含量并计算5种酚酸类成分的积累总量。色谱柱为Inertsil ODS-3 C18色谱柱,流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,使用二极管阵列检测器全波长扫描,流速为1.0 mL·min^(-1),进样量为10μL。并用SPSS 24.0软件进行主成分分析,选取了3个主成分,计算综合得分。结果:上述5种酚酸类成分线性关系良好,相关系数均大于0.998。平均加样回收率介于98.30%~101.22%,RSD为0.43%~2.66%。不同采收期余甘子叶中5种酚酸类成分含量存在一定差异。含量分别为0.2317~1.5074、1.9517~4.6593、2.5105~6.3325、2.2440~5.3386、13.5570~39.1897 mg·g^(-1)。不同采收期5种酚酸类成分总含量变化为7月>5月>8月>6月>1月>9月>4月>2月>10月>12月>3月>11月,以7月份总含量最高,含量达到(55.158±0.83)mg/g,呈现先降后升的动态变化趋势。主成分分析结果表明综合得分最高的月份为7月。结论:建立的方法可用于同时测定不同采收期余甘子叶中5个酚酸类成分的含量。最适宜采收期为7月。

Objective:To establish the method for content determination of the five phenolic acids ingredients of Phyllanthus emblica L.in different harvest time,and to investigate its variation rules so as to determine the best harvest time.Methods:The contents of Epigallocatechin,Corilagin,Scorpion Biphenyl acid,Ellagic acid and Scorpion acid of Phyllanthus emblica L.in different harvest time were determined by HPLC,and the total amount of accumulation of the five active ingredients was calculated.The chromatographic column was Inertsil ODS-3 C18,acetonitrile−0.1%phosphoric acid aqueous solution as mobile phase for gradient elution.A diode array detector was used with full-wavelength scanning.Flow rate was 1.0 mL·min^(−1).Injection was 10μL.Used SPSS 24.0 software for principal component analysis,three principal components were selected and the comprehensive score was calculated.Results:The above five ingredients had a good linear relationship,and the correlation coefficients was more than 0.998,while the average recovery rates were 98.30%~101.22%and RSD were 0.43%~2.66%.There were some differences in the contents of the five phenolic acids ingredients in the leaves of Phyllanthus emblica L.during different harvesting periods.The contents of above five ingredients were 0.2317~1.5074,1.9517~4.6593,2.5105~6.3325,2.2440~5.3386,13.5570~39.1897 mg·g^(−1),respectively.The change in content of the five phenolic acids ingredients in different harvest time was:July>May>August>June>January>September>April>February>October>December>March>November.The total content was the highest in July,which was(55.158±0.83)mg/g and showed a dynamic trend of decreasing first and then increasing.Principal component analysis results showed that the month with the highest comprehensive score was July.Conclusion:Established method can be used to determine the contents of the five phenolic acids ingredients of Phyllanthus emblica L.in different harvest time.July was the best harvest time for Phyllanthus emblica L.