世界中医药

目的:通过高效液相色谱法(HPLC)对20个不同批次同一厂家的六味地黄丸进行指纹图谱的建立,并对其主要成分进行分析。方法:所有20个批次的六味地黄丸均按照统一方法配制供试品溶液。本研究进样体积为2 μL;色谱柱C18限定窄径为2.1 mm,短柱限定为100 mm,填料粒径限定为1.7 μm,温控箱将色谱柱的温度限定为35 ℃。流动相A的主要由0.01%磷酸和超纯水混合而成;流动相B的主要由乙腈和超纯水混合而成,研究中所有高效液相色谱均采用梯度洗脱程序。前期对HPLC法进行常规的方法学考察,当所有方法学考察均可靠良好时,再对所有批次六味地黄丸进行指纹图谱的建立,然后做进一步的相似度分析和主要成分分析。结果:1)六味地黄丸中主要成分梓醇、马钱苷、山药素Ⅰ均可见良好的分离度,最重要的是这3中成分之间并不存在任何内源性的干扰物质,显示各个成分的专属性均较好。2)对照品线性回归分析结果显示:梓醇对照品回归方程:Y=4 110X+798.99;马钱苷对照品回归方程:Y=4 201X-6.013 4;山药素Ⅰ回归方程:Y=2 632X+115.38。3)梓醇、马钱苷和山药素Ⅰ精密度试验的RSD值分别为0.32%、0.51%、0.27%;稳定性试验RSD值分别为1.98%、2.13%、1.11%;重复性试验RSD值分别为1.35%、1.16%、1.02%;加样回收试验的RSD值分别为2.37%、1.54%、1.93%,均符合当RSD值<3%时,说明采用HPLC这一测定方法是可靠的。4)建立六味地黄丸HPLC指纹图谱后,经过相似度计算,从S1-S20的相似度均大于0.90,说明20个批次的六味地黄丸的样品相似度良好。5)20批不同批次的六味地黄丸中中梓醇、马钱苷和山药素Ⅰ的含量差异无统计学意义,20个不同批次的六味地黄丸中梓醇、马钱苷和山药素Ⅰ的含量的平均值分别为(2.176±0.290)mg/g、(1.875±0.119)mg/g、(1.776±0.095)mg/g。结论:采用HPLC建立六味地黄丸指纹图谱,20个不同批次六味地黄丸经过相似度良好,且各主要成分分析间未见明显差异。

English Summary:

To establish the fingerprint of Liuwei Dihuang Pills from 20 different batches of the same manufacturer by high performance liquid chromatography(HPLC)，and to analyze its main components.Methods:All 20 batches of Liuwei Dihuang Pills were prepared into test solutions according to an unified method.In this study，the injection volume was 2 μL，the narrow diameter of column C18 was 2.1 mm，the short column was 100 mm，the particle size of filler was 1.7 μm，and the temperature of column was 35 ℃ in temperature control box.Mobile phase A was mainly composed of 0.01% phosphoric acid and ultra-pure water，while mobile phase B was mainly composed of acetonitrile and ultra-pure water.Gradient elution procedure was used in all the high performance liquid chromatography studies.Previously，the routine methodology of HPLC was investigated.When all the methodological investigations were reliable and good，the fingerprints of all batches of Liuwei Dihuang Pills were established，and then further similarity analysis and principal component analysis were conducted.Results:1)Catalpol，loganin and batatasin I，the main components of Liuwei Dihuang Pills，showed good resolution.Most importantly，there were no endogenous interfering substances among the 3 components，which showed that the specificity of each component was good.2)Linear regression analysis showed that Catalpol regression equation：Y=4110X+798.99; loganin regression equation：Y=4201X-6.013 4; batatasin I regression equation：Y=2632X+115.38.3)RSD values of catalpol，loganin and batatasin I precision test were 0.32%，0.51% and 0.27%; RSD values of stability test were 1.98%，2.13% and 1.11%; RSD values of repeatability test were 1.35%，1.16% and 1.02%; RSD values of sample addition recovery test were 2.37%，1.54% and 1.93% respectively，which were in line with that when RSD values were less than 3%，thus the determination method of HPLC was reliable.4)After establishing the fingerprint of Liuwei Dihuang Pills by HPLC，the similarity calculated from S1-S20 was greater than 0.90，indicating that the samples of 20 batches of Liuwei Dihuang Pills had good similarity.5)The contents of catalpol，loganin and batatasin I in 20 batches of Liuwei Dihuang Pills had no significant statistical difference.The average values of catalpol，loganin and batatasin I in 20 batches of Liuwei Dihuang Pills were(2.176±0.290)mg/g,(1.875±0.119)mg/g,(1.776±0.095)mg/g，respectively.Conclusion:The fingerprint of Liuwei Dihuang Pills was established by HPLC.The similarity of 20 different batches of Liuwei Dihuang Pills was good，and there was no significant difference among the main components.

查看全文查看/发表评论下载PDF阅读器