目的 基于UHPLC-ESI-MS/MS多组分定量技术，系统研究乌头汤制备过程中化学成分的动态变化规律，阐明其量值传递特征，为优化制备工艺提供科学依据。方法 采用Waters Acquity UPLC Hss T3色谱柱（100 mm×2.1 mm，1.8 μm），以0.1%甲酸水溶液-0.1%甲酸乙腈溶液为流动相进行梯度洗脱，结合三重四极杆质谱检测器，建立测定乌头汤中29种化学成分的定量分析方法，并进行方法学验证。基于该方法监测乌头汤现代制备工艺各阶段化学成分的动态变化规律，通过量值传递分析优化乌头汤关键工艺参数。结果 所建立的UHPLC-ESI-MS/MS方法符合《中国药典》2020年版相关要求，29种成分在方法学验证各项检测指标的RSD均小于10.0%。量值传递结果表明，制川乌先煎90 min双酯型生物碱含量波动下降，可有效控制以乌头碱、次乌头碱、新乌头碱为例的双酯型生物碱的总量，而一煎煎煮60 min可显著提高单酯型生物碱及黄酮类成分的溶出，二煎煎煮30 min各化学成分已达最大溶出率。优化后的制备工艺：制川乌单独煎煮90 min后，下麻黄、白芍、黄芪及炙甘草，合煎60 min，药渣再煎30 min。9批验证试验结果显示，测定大部分化学成分含量的RSD值均低于20%，7种微量成分或热敏性成分的RSD值也≤30.5%，表明当前优化工艺基本稳定可控。结论 通过UHPLC-ESI-MS/MS多组分定量技术，系统揭示了乌头汤制备过程中的量值传递规律，并据此优化了关键工艺参数；建立的制备方法可有效控制毒性成分含量，同时保证药效成分的充分溶出，为乌头汤临床安全药效的应用奠定基础。

Objective Based on the UHPLC-ESI-MS/MS multi-component quantitative technology, this study systematically investigates the dynamic changes of chemical components during the preparation of Wutou Decoction (乌头汤), elucidates its quantitative transfer characteristics, and provides a scientific basis for optimizing the preparation process.Methods The analysis was performed using a Waters Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) with a gradient elution of 0.1% formic acid aqueous solution and 0.1% formic acid acetonitrile solution as the mobile phase. Coupled with a triple quadrupole mass spectrometer detector, a quantitative analytical method was established for the simultaneous determination of 29 chemical components in Wutou Decoction. Method validation was also conducted. Based on this method, the dynamic changes of chemical components were monitored across different stages of the modern preparation process of Wutou Decoction. Quantitative transfer analysis was applied to optimize key process parameters.Results The established UHPLC-ESI-MS/MS method complied with the relevant requirements of the Chinese Pharmacopoeia. The RSD values of most of the 29 components in the method validation tests were less than 10.0%. Quantitative transfer analysis revealed that pre-decoction of Zhichuanwu (Aconiti Radix Cocta) for 90 min the content of diester-diterpenoid alkaloids continues to decrease, effectively controlling the total amount of three diester-diterpenoid alkaloids aconitine, hypaconitine, and mesaconitine. Meanwhile, subsequent combined decoction for 60 min significantly enhanced the dissolution of monoester-diterpenoid alkaloids and flavonoids. In the second round of decoction, a 30 min boiling time is sufficient to achieve the maximum dissolution rate for all these chemical components. The preparation process was optimized as follows: Aconiti Radix Cocta was firstly heated for 90 min, then Mahuang (Ephedrae Herba), Baishao (Paeoniae Radix Alba), Huangqi (Astragali Radix) and Zhigancao (Glycyrrhizae Radix et Rhizoma Praeparata cum Melle) were mixed with this Aconiti Radix Cocta solution and further heated for 60 min. After filtration through the gauze, the decocted aqueous solution was collected. The filtered residue was heated for another 30 min. Two decocted aqueous solution was mixed and this was the final Wutou Decoction. Validation tests across nine batches showed that most of the chemical components had the RSD < 20%, while seven trace or heat-sensitive components had RSD ≤ 30.5%, confirming the robustness and controllability of the optimized process.Conclusion This study systematically elucidated the quantitative transfer patterns during the preparation of Wutou Decoction using UHPLC-ESI-MS/MS multi-component quantitative analysis, and further optimized the key process parameters based on these findings. The established preparation method effectively controls the content of toxic constituents while ensuring sufficient extraction of bioactive components, thereby laying a solid foundation for the safe and effective clinical application of Wutou Decoction.

R283.6

国家中医药管理局“国家中药炮制技术传承基地建设项目”（［2022］59号）