复方肾苏Ⅱ的HPLC指纹图谱及化学成分鉴定

HPLC Fingerprint Analysis and Chemical Component Analysis of Traditional Chinese Medicine Compound Shensu Ⅱ

目的：建立复方肾苏Ⅱ（肾苏Ⅱ）HPLC指纹图谱并对其化学成分进行分析,为肾苏Ⅱ质量评价与药效物质基础研究提供依据。方法：采用Waters Symmetry C_（18）色谱柱（4.6 mm×250 mm,5μm）进行检测,以乙腈-水（0.1%H3PO4）为流动相,进行梯度洗脱,流速1.0 mL·min^（-1）,检测波长280 nm,柱温30℃,进样量10μL。测定10批肾苏Ⅱ样品,利用中药色谱相似度评价系统建立肾苏ⅡHPLC指纹图谱,并通过对照品对共有峰进行指认;应用UPLC-ESI-MS/MS技术对肾苏Ⅱ样品中的化学成分进行鉴别。结果：建立了肾苏Ⅱ成分指纹图谱共有模式,显示15个共有峰,各峰分离度良好,各批次间共有峰相对保留时间RSD均〈1.3%,样品间相似度均〉0.9,指认黄芩苷为特征峰。鉴定出肾苏Ⅱ样品中26个化学成分,分别为阿魏酸,汉黄芩素,毛蕊异黄酮,千层纸素,白杨素-6-C-阿拉伯糖-8-C-葡萄糖苷,5,7-二羟基-2＇-甲氧基黄酮-7-O-葡萄糖醛酸苷,5,7,2＇,5＇-四羟基-8,6＇-二甲氧基黄酮,白杨素-6-C-葡萄糖-8-C-阿拉伯糖苷,韧黄芩素Ⅱ,三羟基-单甲氧基黄酮,三羟基-二甲氧基黄酮,芒柄花素,5,7,8-三羟基黄酮,黄芩素,去甲汉黄芩素,黄芩苷,黄芩苷异构体,5,7,8-三羟基-6-甲氧基黄酮-7-O-葡萄糖醛酸苷,白杨素,千层纸素A-7-O-葡萄糖醛酸苷,汉黄芩苷,汉黄芪素,二羟基-二甲氧基黄酮,熊竹素,三羟基-三甲氧基黄酮,黄芩黄酮Ⅱ。结论：该指纹图谱具有良好的精密度、重复性、稳定性,各共有峰间分离度高,可用于肾苏Ⅱ的质量评价;该研究为肾苏Ⅱ物质基础的研究提供基础。

Objective： To establish the HPLC fingerprint of traditional Chinese medicine（ TCM）compound Shensu Ⅱ（ Shensu Ⅱ） and analyze its chemical components for its quality control and medicinal substance study. Method： Waters Symmetry C_（18） column（ 4. 6 mm × 250 mm,5 μm） was used with acetonitrile-0. 1% phosphoric acid solution as the mobile phase for gradient elution. The detective wave length was 280 nm; the column temperature was set at 30 ℃; the injection volume was 10 μL; and the flow rate was 1. 0 mL·min^（-1）. 10 batches of Shensu Ⅱ samples were detected; their HPLC fingerprint was established by using Similarity Evaluation System for Chromatographic Fingerprint of TCM,and common peaks were identified through controls. UPLC-ESIMS/MS was used to analyze the chemical components. Result： Fingerprints of 10 batches of Shensu Ⅱ were established and 15 common peaks were identified. The peaks were well separated,and the relative retention time of common peaks in each batch had RSD of 1. 3% respectively; the similarity was more than 0. 9 among different samples. Baicalin was identified as the characteristic peak,and 26 chemical components were identified as follows：ferulic acid,wogonin,calycosin,oroxylin A,chrysin-6-C-arabinose-8-C-glucoside,5,7-dyhydroxy-2＇-methoxy flavone-7-O-glucuronide, 5, 7, 2＇, 5＇-tetrahydroxy-8, 6＇-dimethoxyflavanone, chrysin-6-C-glucose-8-Carabinoside,tenaxin Ⅱ,trihydroxy-monomethoxy flavone,trihydroxy-dimethoxylflavone,formononetin,5,7,8-trihydroxyflavone,baicalein,norwogonin,baicalin,baicalin isomer,5,7,8-trihydroxy-6-methoxylflavone-7-Oglucuronide, chrysin, qroxylin A-7-O-glucuronide, wogonoside, astragalin, dyhydroxy-dimethoxylflavone,kumatakenin,trihydroxy-trihydroxyflavone,and skullcapflavone Ⅱ. Conclusion： The method can be used for the quality control of Shensu Ⅱ with good precision,high accuracy and reproducibility,providing basis for further studies on therapeutic efficacy of Shensu Ⅱ.