目的 优化赶山鞭总黄酮的提取工艺，建立赶山鞭总黄酮提取物的指纹图谱。方法 通过单因素实验研究提取时间、提取次数、乙醇体积分数、料液比对赶山鞭中总黄酮含量及金丝桃苷的转移率的影响；以此为基础，采用Central-Composite试验设计原理进行3因素5水平试验设计和响应面分析法优化提取工艺。采用比色法，于500 nm处测定药材中以芦丁计总黄酮的含量，采用HPLC法测定总黄酮中金丝桃苷的含量，计算金丝桃苷的转移率。采用HPLC法建立赶山鞭总黄酮提取物的指纹图谱，采用《中药色谱指纹图谱相似度评价软件（2012年版）》进行相似度评价，确定共有峰，并采用SPSS 22.0软件进行聚类分析和主成分分析。结果 赶山鞭总黄酮的最佳提取条件为以65%乙醇，料液比1∶10，提取120 min，提取2次。建立的赶山鞭药材总黄酮提取物的HPLC指纹图谱有15个共有指纹峰，指纹图谱的相似度分析结果显示相似度均大于0.61，表明10批样品的化学成分一致性较好，但各成分含量存在差异，对部分色谱峰进行了归属指认分别是金丝桃苷、槲皮素、芦丁、山柰酚。聚类分析可知欧氏距离为25时，10批样品可聚为2类，S1与S7为一类，其余聚为一类；欧氏距离为15时，可聚为5类，即S3、S5、S6、S8～S10聚为一类，S1、S2、S4、S7各为一类。经主成分分析，4个主成分因子的累积方差贡献率为88.659%，以S5样品的主成分因子综合得分最高。结论 响应面法优化的赶山鞭总黄酮提取纯化条件简单可靠，建立的HPLC指纹图谱能够简便、高效地反映赶山鞭总黄酮提取物中大部分化学成分信息，为赶山鞭药材的质量评价奠定基础。

Objective To optimize the extraction process of the total flavonoids of Ganshanbian (Hypericum attenuatum) and to establish the fingerprint profile of the total flavonoids of H. attenuatum. Methods Single-factor experiments were conducted to study the effects of solvent extraction time, extraction times, extraction concentration, and material-to-liquid ratio on the extraction rate and transfer rate of hyperoside of total flavonoids in H. attenuatum, on this basis, the central- composite design experimental design principle is used to carry out a three-factor five-level experimental design and response surface analysis to optimize the extraction process; on the basis of obtaining the best extraction method, on the basis of the best extraction method, the purification method of the total flavonoids extract of H. attenuatum was optimized by using the type of resin, the loading concentration of the extract, the loading volume of the extract, the elution concentration and the elution volume as the investigating indexes. After that, the fingerprints of the total flavonoids extract of H. attenuatum were established by HPLC, and SPSS 22.0 software was used for cluster analysis and principal component analysis. Results The best extraction conditions were 80% ethanol, 1:10 ratio, 2 h per extraction, extract 2 times. The HPLC fingerprints of the total flavonoids of H. attenuatum were determined to have 15 peaks common to all chromatograms, and the similarity analysis of the fingerprints showed that the similarity was greater than 0.61, the peaks were identified as auriculoside, quercetin, rutin and sannaphene, indicating that the chemical composition of the 10 batches of samples was consistent, but there were differences in the content of each component. The clustering analysis showed that when the Euclidean distance was 25, the 10 batches of samples could be clustered into 2 classes, S1 and S7 into one class, and the rest into one class; when the Euclidean distance was 5, they could be clustered into 5 classes, namely S3, S5, S6, S8-S10 into one class, and S1, S2, S4, S7 into one class each. After the principal component analysis, the cumulative variance contribution of the four principal component factors was 88.659%, with the highest overall score of the principal component factors for S5 samples.Conclusions The extraction and purification conditions of the total flavonoids of H. attenuatum were optimized by response surface analysis. The HPLC fingerprint was able to easily and efficiently reflect most of the chemical information in the extract of H. attenuatum Choisy, which could provide reference for the quality evaluation and related study of H. attenuatum.

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国家自然科学基金面上项目（82174232）；黑龙江省自然科学基金优秀青年项目（YQ2019H031）；黑龙江中医药大学“优秀创新人才支持计划”项目（2018年）；黑龙江省博士后研究人员落户黑龙江科研启动资助项目（2020年）