目的 建立百药煎HPLC指纹图谱，比较五倍子发酵百药煎化学成分变化，为百药煎的质量评价提供方法。方法 采用反相高效液相色谱（RP-HPLC）法，Waters Symmmetry Shield^TM RP₁₈（250 mm×4.6 mm，5 μm）C₁₈色谱柱，乙腈-0.1%三氟乙酸水溶液梯度洗脱，体积流量0.8 mL/min，检测波长280 nm，"Chromap Chromafinger 2005 beta 0.1"色谱软件生成百药煎标准指纹图谱，对10批不同批次的百药煎化学成分指纹图谱进行相似度计算，并且通过对照品比对及高效液相色谱与质谱联用技术（HPLC-MS）对主要共有峰进行指认。结果 10批百药煎指纹图谱中有10个共有峰，HPLC指纹图谱各峰分离度良好，各批次间共有峰的相对保留时间RSD均<1.0%，样品间相似度均>0.9，共指认出7个共有峰，即没食子酸（1号峰）、表没食子儿茶素（2号峰）、没食子酸甲酯（3号峰）、没食子酸乙酯（5号峰）、表没食子儿茶素没食子酸酯（6号峰）、2，4，6-三-O-没食子酰-α-D-葡萄糖（7号峰）、2，4，6-三-O-没食子酰-β-D-葡萄糖（9号峰）；五倍子发酵百药煎后没食子酸、2，4，6-三-O-没食子酰-α-D-葡萄糖的量升高，没食子酸甲酯、没食子酸乙酯、表没食子儿茶素没食子酸酯的量明显降低；表没食子儿茶素、2，4，6-三-O-没食子酰-β-D-葡萄糖为新生成成分。结论 百药煎炮制作用机制与发酵过程使五倍子化学成分发生变化和产生新的化学成分有关。指纹图谱法可用于监控百药煎发酵炮制的质量控制。

Objective To study and establish the HPLC fingerprint standard for the quality analysis and compare effects on the chemical composition of gallnut by ferment of Chinese gall leaven. Methods The fingerprint of Chinese gall leaven was built by Waters Symmmetry Shield^TM RP₁₈ (250 mm×4.6 mm, 5 μm) C₁₈ column, and acetonitrile-0.1% trifluoroacetic acid aqueous in gradient as mobile phase, the flow rate was 0.8 mL/min, and the detecting wavelength was set at 280 nm. The chemical fingerprint similarity of 10 batches of Chinese gall leaven was calculated with the Chromap Chromafinger 2005 beta 0.1 standard substance comparison and HPLC-MS were adopted to identify the common peaks. Results The fingerprint chromatography for the 10 batches of Chinese gall leaven included 10 common peaks, with a good separation at each peak. The relative retention time for common peaks of each batch was less than 1.0%, and the similarities among 10 samples were greater than 0.90. Gallic acid (peak 1), (-)-epigallocatechin (EGC, peak 2), methyl gallate (peak 3), ethyl gallate (peak 5), epigallocatechin gallate (EGCG, peak 6), 2,4,6-tri-O-galloyl-β-D-glucose (peak 7), and 2,4,6-tri-O-galloyl-α-D-glucose (peak 9) were identified. The gallnut fermented made the content of gallic acid and 2,4,6-tri-O-galloyl-α-D-glucose increased and the contents of methyl gallate, ethyl gallate, and (-)-epigallocatechin gallate decreased. It was found that (-)-epigallocatechin and 2,4,6-tri-O-galloyl-β-D-glucose had formed in the process for the first time. Conclusion The processing mechanism of Chinese gall leaven is related to gallnut fermentation process change and create new chemical composition and fingerprint can be used to monitor the quality of fermentation processing of Chinese gall leaven.

公益性行业专项：六神曲等7种中药发酵技术及规范化应用研究——百药煎（201507004-03）