断血流胶囊化学成分、色谱指纹图谱及含量测定的研究

Chemical Compositions,Chromatographic Fingerprints and Content Determination of Duanxueliu Capsules

目的建立断血流胶囊高效液相色谱(HPLC)指纹图谱及多成分含量测定方法,为其质量评价提供参考。方法基于纳升高效液相色谱-四极杆-静电场轨道阱超高分辨质谱(Q-Exactive Plus Orbitrap MS)技术分析断血流胶囊化学成分;采用HPLC法,建立12批断血流胶囊样品HPLC指纹图谱,以《中药色谱指纹图谱相似度评价系统》(2012版)评价指纹图谱相似度,并结合主成分分析(PCA)及偏最小二乘法判别分析(PLS-DA)筛选差异标志物;同时测定没食子酸、木犀草素和醉鱼草皂苷Ⅳb含量。结果从断血流胶囊中鉴定出36个成分。12批样品确定26个共有峰,相似度均大于0.90;指认了没食子酸、芦丁、橙皮苷、槲皮素、木犀草素和醉鱼草皂苷Ⅳb 6个共有峰;12批样品按不同生产厂家可分为3类,筛选出木犀草素、峰21、醉鱼草皂苷Ⅳb为差异标志物。12批样品中没食子酸、木犀草素、醉鱼草皂苷Ⅳb质量分数范围分别为0.17~1.24 mg·g^(-1)、0.54~1.27 mg·g^(-1)、16.41~20.98 mg·g^(-1)。各成分的线性关系良好(r≥0.9998),平均回收率为99.6%~100.6%。结论明确了断血流胶囊的化学成分,所建立的HPLC指纹图谱及多成分含量测定方法稳定、可靠,可为断血流胶囊的质量评价提供依据。

Objective To establish HPLC fingerprints and multi-compositions content determination method for Duanxueliu Capsules,and provide reference for the quality evaluation of Duanxueliu Capsules.Methods Analysis and identification of the chemical compositions of Duanxueliu Capsules based on nanoliter high performance liquid chromatography-quadrupole-electrostatic field orbitrap ultra-high resolution mass spectrometry(Q-Exactive Plus Orbitrap MS)were carried out.HPLC method was used to establish fingerprints of 12 batches of samples.The similarity evaluation was conducted by using Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 Edition).Principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA)were performed to screen the markers for difference.The contents of 3 compositions,gallic acid,luteolin,and buddlejasaponinⅣb were simultaneously determined by HPLC.Results Thirty-six compositions were identified in Duanxueliu Capsules.The similarities of 12 batches of Duanxueliu Capsules were greater than 0.90,and six common peaks were identified,including gallic acid,rutin,hesperidin,quercetin,luteolin,and buddlejasaponinsⅣb.The 12 batches of samples analyzed by PCA were divided into 3 categories according to different manufacturers.Differential markers were identified based on PCA,including luteolin,peak 21,and buddlejasaponinsⅣb.The contents of gallic acid,luteolin,and buddlejasaponins IVb in 12 batches of Duanxueliu Capsules were 0.17-1.24 mg·g^(-1),0.54-1.27 mg·g^(-1),and 16.41-20.98 mg·g^(-1),respectively.The linear relationships between 3 compositions were good(r≥0.9998),the average recoveries were 99.6%-100.6%.Conclusion Chemical compositions of Duanxueliu Capsules were identified,and the established HPLC fingerprints and multi-compositions content determination method were stable and reliable,which could provide a basis for the quality evaluation of Duanxueliu Capsules.