目的 建立正骨丸的指纹图谱及含量测定方法，阐释正骨丸的入血成分及其药效物质基础，为其质量控 制与药效机制研究提供依据。方法 采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF/MS）技术对 正骨丸水煎液及给药后血浆样品进行成分分析，筛选关键入血成分。基于关键入血成分，优化色谱条件构建正 骨丸高效液相色谱（HPLC）指纹图谱，并结合外标法建立多指标含量测定方法。结果 正骨丸中鉴定出 43 个 成分，主要为黄酮、苯丙素、异喹啉和萜类成分。在大鼠血清中鉴定出芍药苷、洋川芎内酯 I、丹皮酚、地黄 苷 D 和补骨脂素 5 种原型入血成分；建立的指纹图谱共标定 35 个共有峰，10 批样品相似度均＞0.98。多指标 含量测定方法的平均加样回收率范围为 97.09%～98.77%，10 批样品中芍药苷、洋川芎内酯 I、补骨脂素、丹 皮酚、地黄苷 D 含量范围分别为 1.303～1.435 、0.109～0.141、0.103～0.116、0.969～1.028、0.386～0.416 mg·g -1 。 结论 芍药苷、洋川芎内酯 I 和丹皮酚等 5 种入血成分为正骨丸的潜在药效物质基础。UPLC-Q-TOF/MS 技术 结合指纹图谱与多指标成分含量测定方法可为正骨丸质量控制及后续药效研究提供技术支持。

Objective To elucidate the blood-entering components and pharmacodynamic material basis of Bone- Setting Pills，and to establish a fingerprint and content determination method based on these components，so as to provide a basis for its quality control and pharmacological mechanism research. Methods Ultra performance liquid chromatography-quadrupole time of flight tandem mass spectrometry （UPLC-Q-TOF/MS） technology was used to analyze the components of the Bone-Setting Pills and plasma samples after administration，in order to screen for key blood- entering components. For the key blood-entering components，chromatographic conditions were optimized to establish an HPLC fingerprint of Bone-Setting Pills，and a multi-component quantitative determination method was established in combination with the external standard method. Results Forty-three components were identified from Bone-Setting Pills， mainly flavonoids， phenylpropanoids， isoquinolines， and terpenoids. Five prototype blood-entering components were identified in rat serum：paeoniflorin，senkyunolide I，paeonol，rehmannioside D，and psoralen. The established fingerprint chromatogram contained 35 common peaks，and the similarity of 10 batches of samples was all ＞0.98. The average recovery rates of the multi-index content determination method ranged from 97.09% to 98.77%. The content ranges of paeoniflorin，senkyunolide I，psoralen，paeonol，and rehmannioside D in 10 batches of samples were 1.303-1.435，0.109-0.141，0.103-0.116，0.969-1.028，and 0.386-0.416 mg·g -1 ，respectively. Conclusion The five blood-entering components， including paeoniflorin， senkyunolide I， and paeonol， constitute the pharmacodynamic material basis of Bone-Setting Pills. The UPLC-Q-TOF MS technology combined with fingerprint and multi-component quantitative determination can provide technical support for the quality control and subsequent efficacy research of Bone-Setting Pills.

R284.1

国家自然科学基金青年科学基金项目（82205142）；江苏省中医退行性骨关节病临床医学创新中心资助项目（苏中医科教〔2023〕1号）； 无锡市卫生健康委中青年拔尖人才资助计划项目（BJ2023072）；无锡市中药管理局项目（ZYYB20）。