骨疏灵的UPLC指纹图谱的建立及8种成分的含量测定  

UPLC fingerprints of Gushuling and content determination of 8 components

作　　者：常红 袁腾腾 吕淑婕 王雷 陈卫东 张彩云 桑冉[3,4] CHANG Hong;YUAN Teng-teng;LYU Shu-jie;WANG Lei;CHEN Wei-dong;ZHANG Cai-yun;SANG Ran(College of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012;Anhui Key Laboratory of Chinese Medicinal Formula,Hefei 230012;The First Affiliated Hospital of Bengbu Medical College,Bengbu Anhui 233099;Bengbu Medical College,Bengbu Anhui 233030)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]安徽省中药方剂重点实验室,合肥230012 [3]蚌埠医学院第一附属医院,安徽蚌埠233099 [4]蚌埠医学院,安徽蚌埠233030

出　　处：《中南药学》2022年第1期159-164,共6页Central South Pharmacy

基　　金：安徽高校自然科学研究项目(No.KJ2019A0314)。

摘　　要：目的采用超高效液相色谱法(UPLC)建立骨疏灵(GSL)的指纹图谱并测定其中8种成分的含量。方法色谱柱为ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7μm);流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱;柱温为35℃;流速为0.15 mL·min^(-1);进样量为2μL;检测波长为260 nm。使用《中药色谱指纹图谱相似度评价系统(2012版)》建立GSL的指纹图谱,与对照品比对指认出色谱峰,并进行相似度评价。使用SPSS 26.0软件对12批样品进行聚类分析。同时测定12批GSL中淫羊藿苷等8种成分的含量。结果标定了共有峰43个,指认出峰13为毛蕊异黄酮-7-O-葡萄糖苷、峰16为蜕皮激素、峰28为芒柄花苷、峰30为毛蕊异黄酮、峰37为朝藿定A、峰38为朝藿定B、峰41为朝藿定C、峰42为淫羊藿苷。12批GSL指纹图谱与对照指纹图谱的相似度为0.9920~0.9960。当平方欧氏距离为2时,12批样品聚为三类,S4聚为一类,S7聚为一类,其余10批聚为一类;当平方欧氏距离为3时,12批样品可聚为两类,S4聚为一类,其余11批聚为一类。测定的8种成分的含量分别为0.931~1.985、2.197~5.058、0.319~0.737、0.134~0.325、3.380~6.770、17.463~35.547、18.298~37.390、11.933~24.368 mg·g^(-1)。结论首次建立了骨疏灵UPLC指纹图谱结合含量测定的质量控制方法,可为该制剂物质基准的质量控制与评价提供参考。Objective To establish the UPLC fingerprint of Gushuling(GSL)and determine the content of 8 components in the preparation.Methods The chromatographic separation was performed on ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7μm)column with gradient elution.The mobile phase consisted of 0.1%formic acid aqueous solution(A)-acetonitrile(B)at 35℃.The flow rate was 0.15 mL·min^(-1).The injection volume was 2μL.The detection wavelength was 260 nm.The fingerprints of 12 batches of GSL were established by Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition).The chromatographic peaks were identified by comparison with the reference substance,and the similarity was evaluated.SPSS 26.0 software was used for cluster analysis of 12 batches of samples.The content of 8 components such as icariin in the 12 batches of GSL were determined by UPLC.Results Totally 43 peaks were identified.Peak 13,16,28,30,37,38,41 and 42 were calycosin-7-glucoside,ecdysterone,ononin,calycosin,epimedin A,epimedin B,epimedin C,and icariin.The similarity between the 12 batches of GSL fingerprints and the control fingerprints ranged from 0.9920 to 0.9960.When the square Euclidean distance was 2,the 12 batches of samples were clustered into 3 groups:S4 in one group,S7 in one group,and other 10 batches in another group.When the square Euclidean distance was 3,the 12 batches of samples were clustered into 2 categories:S4 in one group,and other 11 batches in other group.The content of the above 8 components were 0.931~1.985,2.197~5.058,0.319~0.737,0.134~0.325,3.380~6.770,17.463~35.547,18.298~37.390,and 11.933~24.368 mg·g^(-1),respectively.Conclusion The quality control method of Gushuling UPLC fingerprint combined with content determination is established for the first time,which can provide a reference for the quality control and evaluation of the preparation substance standard.

关 键 词：骨疏灵 超高效液相色谱法 指纹图谱 聚类分析 含量测定 

分 类 号：R285[医药卫生—中药学] R917[医药卫生—中医学]