HPLC-DAD-ESI-TOF/MS鉴别和测定前列欣胶囊中多种成分及其多指标定量指纹图谱研究

目的：高效液相色谱-电喷雾飞行时间质谱（HPLC-ESI-TOF/MS）联用技术用于前列欣胶囊多种化学成分快速鉴定和测定及多指标定量指纹图谱研究。方法：采用Kromasil 100-5C₁₈（4.6 mm ×250 mm，5μm）色谱柱，以乙腈-0.8%甲酸流动相体系进行梯度洗脱，流速0.8 mL·min^-1，柱温25 ℃，检测波长300 nm；利用ESI-TOF/MS法对前列欣胶囊提取物中多种化合物进行鉴别，对已知的活性成分进行定量测定，并建立前列欣胶囊的HPLC指纹图谱。结果：应用HPLC-DAD-ESI-TOF/MS指认了前列欣胶囊提取物中的8个成分（没食子酸、绿原酸、咖啡酸、王不留行黄酮苷、异槲皮苷、丹酚酸B、丹酚酸A、隐丹参酮），定量测定结果表明各化合物线性关系良好（r=0.999 0~0.999 7）；平均回收率分别为95.25%、96.85%、98.10%、96.71%、97.87%、95.80%、97.57%和96.74%。10批样品中上述各化合物的质量分数分别为194.39~209.43、242.65~253.32、82.79~86.46、132.45~146.80、169.08~188.13、402.36~429.87、595.75~637.11、112.78~122.92 μg·g^-1；10批样品的相似度在0.999 6~1.000 0之间。结论：本研究所建立的活性成分快速鉴别和多指标定量指纹图谱测定方法，为提高前列欣胶囊的质量控制水平提供了技术支持。

Objective:To establish a high performance liquid chromatography-electrospray time-of-flight mass spectrometry(HPLC-ESI-TOF/MS) method for rapid identification and determination of multiple components in Qianliexin capsules,and to establish the quantitative fingerprint. Methods:Separation was carried on Kromasil 100-5 C₁₈ column using acetonitrile-0.8% formic acid as mobile phase at the flow rate of 1 mL·min^-1 for gradient elution and the detection wavelength was 300 nm. The column temperature was set at 25 ℃. ESI-TOF/MS was used for identification of chemical ingredients in Qianliexin Capsules. Assay of multi-components in Qianliexin capsules and its fingerprint were established. Results:Eight ingredients (gallic acid,chlorogenic acid,caffeic acid,vaccarin,isoquercitrin,salvianolic acid B,salvianolic acid A and cryptotanshinone) were identified by the developed HPLC-DAD-ESI-TOF/MS method. Results of assay showed that good linearity (r=0.999 0-0.999 7) was found for the 8 compounds. And the average recoveries of gallic acid,chlorogenic acid,caffeic acid,vaccarin,isoquercitrin,salvianolic acid B,salvianolic acid A and cryptotanshinone were 95.25%,96.85%,98.10%,96.71%,97.87%,95.80%,97.57% and 96.74%,respectively. The contents of the above compounds in 10 samples were in the ranges of 194.39-209.43 μg·g^-1,242.65-253.32 μg·g^-1,82.79-86.46 μg·g^-1,132.45-146.80 μg·g^-1,169.08-188.13 μg·g^-1,402.36-429.87 μg·g^-1,595.75-637.11 μg·g^-1 and 112.78-122.92 μg·g^-1,respectively. And the similarity values of 10 batches of samples ranged from 0.999 6 to 1.000 0. Conclusion:The proposed method for rapid identification and multi-component quantitative fingerprint provided technology support for the quality evaluation of Qianliexin capsules.