目的 探究何首乌Polygoni Multiflori Radix采用古法黑豆"九蒸九晒"和现代药典黑豆汁连续蒸制对主要药效成分含量的影响。方法 模拟"九蒸九晒"的时间安排，制备古法黑豆与何首乌分层铺放蒸后晒品、拌入黑豆汁蒸后晒品和药典拌入黑豆汁后连续蒸制品；建立超高压液相三重四极杆质谱（UPLC-QqQ-MS/MS）测定何首乌中12个成分的分析方法并进行检测；结合t检验、主成分分析和正交偏最小二乘法-判别分析评价不同炮制方法所得样品的差异。结果 建立了可靠的UPLC-QqQ-MS/MS测定何首乌中大黄素、大黄素甲醚、大黄酸、大黄素-8-O-β-D-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、顺式-2，3，5，4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（cis-THSG）、反式-2，3，5，4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（trans-THSG）、虎杖苷、白藜芦醇、表儿茶素、芦丁和金丝桃苷含量的方法；随着蒸制时间的延长，12个成分的含量发生了明显变化：游离蒽醌类成分先降低后升高，蒽醌苷类成分、cis-THSG、虎杖苷和金丝桃苷先升高后降低，trans-THSG、白藜芦醇、表儿茶素和芦丁降低；成分的含量与蒸制用的辅料和操作方法及炮制时间密切相关，操作方法的影响大于辅料对何首乌质量的影响。结论从成分含量的角度分析，古法黑豆蒸晒与药典法的黑豆汁连续蒸制不能等同，该研究为传承和发扬何首乌的传统炮制方法提供了实验依据。

Objective To explore the influence of ancient processing method black bean "nine cycles of steaming and drying" and modern pharmacopoeia processing method continuous steaming with black bean decoction on the main components of Polygoni Multiflori Radix (PMR). Methods Simulating the time arrangement of "nine cycles of steaming and drying", samples were prepared using three processing methods:ancient method that raw PMR (rPMR) and black bean were steamed in layers and then dried, modern method that rPMR were steamed with black bean decoction and then dried, the method recorded in Chinese Pharmacopoeia that rPMR were steamed continuously with black bean decoction; The determination method of 12 components in rPMR and processed PMR (pPMR) was established using ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS), and 12 components in all samples processed by different methods were determined; The results was analyzed combining with t test, principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). Results A reliable UPLC-QqQ-MS/MS method was established for the determination of emodin, physcion, rhein, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside, cis-2,3,5,4'-tetrahydroxylstilbene-2-O-β-D-glucoside (cis-THSG), trans-2, 3,5,4'-tetrahydroxylstilbene-2-O-β-D-glucoside (trans-THSG), polydatin, resveratrol, epicatechin, rutin and hyperoside. With the prolongation of steaming time, the content of 12 effective components changed obviously:the content of free anthraquinone was decreased first and then increased; The content of anthraquinone glycoside, cis-THSG, polydatin and hyperoside was increased first and then decreased; The content of trans-THSG, resveratrol, epicatechin and rutin was decreased; The components content were closely related to the auxiliary materials, steaming operation methods and processing time, the influence of operation methods was greater than that of auxiliary materials on the quality of pPMR. Conclusion The ancient processing method steaming with black bean and drying could not be equated with the modern pharmacopoeia processing method continuous steaming with black bean decoction in terms of the content of effective components. The result provides experimental basis for inheriting and developing the traditional processing method of PMR.

R286.02

上海中医药大学研究生创新创业培养项目（Y2019089）；上海市大学生创新活动计划项目（2019SHUTCM066）；上海市大学生创新活动计划项目（2020SHUTCM048）