目的 建立三黄片（Sanhuang Tablets）UPLC多组分定量分析方法，联合化学模式识别分析及优劣解距离法（technique for order preference by similarity to an ideal solution，TOPSIS）分析，对不同厂家的三黄片质量进行综合评价，为三黄片的品质评价和质量控制提供参考依据。方法 以Waters Acquity UPLC Hss T3（150 mm×2.1 mm，1.8 µm）柱为色谱柱，以甲醇-0.1%磷酸水溶液为流动相，梯度洗脱，检测波长为254、276 nm，UPLC同时检测芦荟大黄素-8-O-葡萄糖苷、大黄酚-1-O-葡萄糖苷、大黄酚-8-O-葡萄糖苷、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、黄芩苷、千层纸素A-7-O-β-D-葡萄糖醛酸苷、汉黄芩苷、黄芩素、汉黄芩素、没食子酸14种成分及盐酸小檗碱的含量，对抽样于7个厂家的52批次三黄片进行含量测定；利用SIMCA 14.1和SPSS 25.0软件对上述15种成分含量检测结果进行层次聚类分析（hierarchical cluster analysis，HCA）、主成分分析（principal component analysis，PCA）和正交偏最小二乘法-判别分析（orthogonal partial least squares-discriminant analysis，OPLS-DA），挖掘影响三黄片质量的关键成分；运用TOPSIS对7个厂家三黄片质量的优劣排序，建立三黄片质量综合评价方法。结果 三黄片中UPLC定量分析方法学考察均符合《中国药典》2020年版规定要求，精密度、稳定性、重复性及准确度的RSD值均小于3.0%。HCA和PCA结果均显示52批次三黄片聚为3类；PCA共提取到4个主成分，累积方差贡献率为91.309%；OPLS-DA筛选出变量重要性投影（variable importance in projection，VIP）值＞1.0的6个差异性标志物大黄素甲醚、没食子酸、盐酸小檗碱、大黄酚、黄芩苷、芦荟大黄素-8-O-葡萄糖苷，可能是影响三黄片质量的主要成分；TOPSIS欧氏贴近度在0.184 9～0.556 6，表明不同厂家间三黄片质量差异较大，其中厂家A、B的排名靠前。结论 建立的多成分定量检测方法操作便捷，能准确可靠的测定三黄片中15种化学成分含量；HCA、PCA和OPLS-DA可有效筛选出影响三黄片质量的差异性标志物，联合TOPSIS法可用于综合评价三黄片质量。

Objective To establish a multi-component quantitative method for the UPLC analysis of Sanhuang Tablets (三黄片, SHT), combined with chemical pattern recognition analysis and technique for order preference by similarity to an ideal solution (TOPSIS) analysis, to comprehensively evaluate the quality of SHT from different manufacturers, and to provide a reference basis for the quality evaluation and quality control of SHT. Methods The separation was performed on a Waters Acquity UPLC Hss T3 (150 mm×2.1 mm, 1.8 µm) column with methanol-0.1% aqueous phosphoric acid as the mobile phase in a gradient elution, and the detection wavelengths were 254 nm and 276 nm. UPLC was used for the simultaneous determination of content of aloe-emodin-8-O-glucopyranoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucopyranoside, aloe-emodin, rhein, emodin, chysophanol, physcion, baicalin, oroxylin A-7-O-β-D-glucuronide, wogonoside, baicalein, wogonin, gallic acid and berberine hydrochloride, and the content of 52 batches of SHT sampled from seven manufacturers were determined; HCA, PCA and OPLS-DA were performed on the content test results of the above 15 components using SIMCA 14.1 and SPSS 25.0 software to dig out the key components affecting the quality of SHT; TOPSIS was used to rank the advantages and disadvantages of SHT quality of seven manufacturers, and establishing a comprehensive evaluation method of SHT quality. Results The methodological investigation of UPLC quantitative analysis in SHT meets the requirements specified in the 2020 edition of the Chinese Pharmacopoeia. The RSD values of precision, stability, repeatability and accuracy were less than 3.0%. The results of both HCA and PCA showed that the 52 batches of SHT were clustered into three classes; a total of four principal components were extracted from the PCA, and the cumulative variance contribution rate was 91.309%; OPLS-DA screened six differential markers with variable importance in projection (VIP) values >1.0, including physcion, gallic acid, berberine hydrochloride, chysophanol, baicalin, aloe-emodin-8-O-β-D-glucopyranoside, which were the main components affecting the quality of SHT; TOPSIS Euclidean closeness ranges from 0.184 9 to 0.556 6, indicating that the quality of SHT varies greatly among different manufacturers, with manufacturers A and B ranking high. Conclusion The established multi-component quantitative assay is easy to operate and can accurately and reliably determine the contents of 15 chemical components in SHT; HCA, PCA and OPLS-DA can effectively screen out the difference markers influencing the quality of SHT, which combined with TOPSIS method can be used to comprehensively evaluate the quality of SHT.

R283.6

国家重点研发计划——中医药现代化专项（2023YFC3504101）