温胆汤UPLC-UV-MS/MS指纹图谱研究

目的 研究和建立温胆汤UPLC-UV指纹图谱,确定色谱峰药材来源,同时对各成分进行定性研究,为该方物质基础和质量控制提供科学依据.方法 UPLC-UV法进行温胆汤指纹图谱研究,对14批样品进行相似度分析;阴性液对照法进行共有峰药材来源研究;UPLC-MS/MS法对温胆汤中成分进行定性研究.结果 建立了温胆汤UPLC-UV指纹图谱,进行了方法学考察,结果显示该方法精密度、稳定性、重复性良好;对14批样品进行指纹图谱研究,得出共有峰指纹图谱,标记了22个共有峰;进行了14批样品相似度比较,相似度在0.863～0.998;通过与对照品保留时间比较,直接指认了橙皮苷、新橙皮苷、柚皮苷、甘草苷和甘草酸铵;与药材阴性液比较,对22个共有峰药材来源进行了归属;UPLC-MS/MS法定性了橙皮苷、新橙皮苷、柚皮苷、甘草苷和甘草酸铵5个成分,推断出了圣草枸橼苷、异柚皮苷、枸橘苷、乌拉尔甘草皂苷乙4个成分.结论 建立的指纹图谱方法准确、可靠,对温胆汤物质基础和质量控制研究提供了一定参考.

Fingerprint study of Wendan Decoction by UPLC-UV-MS/MS

Objective To study and establish the UPLC fingerprint of Wendan Decoction (WDD), to ascertain the medicinal source of chromatographic peaks, and to identify the chemical constituents for providing scientific basis in effective substance foundation and quality control of WDD. Methods The fingerprint of WDD was studied by UPLC-UV, and the similarity threshold of 14 batches of samples of WDD was analyzed; The medicinal source of commom peaks was studied by negative control method; UPLC-MS/MS method was used for the qualitative analysis on chemical constituents in WDD. Results The UPLC fingerprint of WDD was established, the methodology was examined, and the method showed good precision, stability, and repeatability. The fingerprints of 14 batches of sample were studied and 22 common peaks were signed. The similarity threshold was compared, and it was between 0.863-0.998. Five constituents were identified as hesperidin, neohesperidin, naringin, liquiritin, and glycyrrhizic acid by comparing the retention time and ultraviolet spectra of the peaks with those of the reference substances. The attribution of 22 common peaks was analyzed by comparing with negative control. UPLC-MS/MS method was used for the qualitative analysis on hesperidin, neohesperidin, naringin, liquiritin, and glycyrrhizic acid and inferring eriocitrin, isonaringin, poncirin, and uralsaponin B. Conclusion The method is stable and reliable, providing a certain reference to substance basis of WDD.