摘要
目的:建立青白通痹胶囊的指纹图谱,结合化学计量学和网络药理学方法,分析预测青白通痹胶囊潜在的质量标志物(Q-Marker),并对Q-Marker成分进行含量测定,为其质量控制提供参考。方法:采用Agilent 5 TC-C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),柱温30℃,进样体积10μL,紫外检测波长210 nm(0~45 min)、260 nm(45~70 min),建立指纹图谱并确定共有峰,通过与对照品相对保留时间及紫外光谱的比对进行色谱峰对应化合物的指认,经单煎液对各共有峰进行归属。采用化学计量法对10批青白通痹胶囊进行质量评价,借助正交偏最小二乘-判别分析(OPLS-DA)分析筛选青白通痹胶囊的主要差异性标志物;结合网络药理学,通过相应数据库筛选差异性标志物的核心靶点和关键通路,构建“成分-靶点-通路”网络图,结合上述结果筛选可用于预测青白通痹胶囊的Q-Marker,并建立对预测得到的标志性成分进行含量测定的HPLC方法。结果:建立了青白通痹胶囊的HPLC指纹图谱,标记24个共有峰,对其进行峰归属,其中1~3、5~8、10~11、13、16号峰来自青风藤,4、9、12、14、16、18~19号峰来自白芍,15、17、20~24号峰来自炙甘草,并指认了其中的8个共有峰,分别为儿茶素、青藤碱、没食子酸、木兰花碱、芍药苷、甘草苷、1,2,3,4,6-O-五没食子酰葡萄糖、甘草酸。相似度评价显示,10批青白通痹胶囊样品的相似度为0.934~1.000。主成分分析(PCA)显示前4个主成分的累积方差贡献率为93.998%,OPLS-DA显示有8个成分变量重要性投影(VIP)值较大。采用网络药理学的方法分析得出儿茶素、青藤碱等活性成分可能通过37个核心靶点、10条主要通路发挥药效作用,初步筛选儿茶素、青藤碱、没食子酸、木兰花碱、芍药苷、甘草苷、1,2,3,4,6-O-五没食子酰葡萄糖、甘草酸为青白通痹胶囊潜在的Q-Marker。同时测定这8个成分的含量,方法学考察结果良好,平均加样回收率为96.81%~103.44%,RSD为0.6%~3.7%。10批样品中儿茶素、青藤碱、没食子酸、木兰花碱、芍药苷、甘草苷、1,2,3,4,6-O-五没食子酰葡萄糖、甘草酸的质量分数分别为0.9071~1.1893、2.1833~3.1186、0.3970~1.4276、3.5079~5.4466、14.2077~19.5701、1.4128~3.5775、0.4420~1.6977、2.7388~4.7612 mg·g^(-1)。结论:本研究建立的青白通痹胶囊指纹图谱方法简单,重复性好,筛选的指标性成分将为青白通痹胶囊的质量控制提供依据。
Objective:To establish the fingerprint of Qingbai Tongbi capsules,and to screen out its indicative compounds for quality control combined with chemometrics methods and network pharmacology.Methods:Agilent 5 TC-C_(18)(250 mm×4.6 mm,5μm)chromatographic column was used for separation.The mobile phase was acetonitrile-0.1%phosphoric acid solution for gradient elution at a flow rate of 1.0 mL·min^(-1),and the column temperature was 30℃.The injection volume was 10μL,and the wavelength was 210 nm(0-45 min),260 nm(45-70 min).The fingerprint was established and the common peaks were determined.By comparing with the relative retention time and UV spectra of the reference substances,the corresponding compounds of the chromatographic peak were identified.Then the common peaks were identified by single decoction.Chemometrics methods was used to evaluate quality of 10 batches of Qingbai Tongbi capsules,and OPLS-DA analysis was used to screen out the main marker components of Qingbai Tongbi capsules.Combined with network pharmacology,the core targets and key pathways were constructed a"component-target-pathway"network map through corresponding databases.Combined with the above results,indicative compounds for quality control of Qingbai Tongbi capsules were screened out,and an HPLC method was established to determine the content of the Q-Markers.Results:An HPLC fingerprint of Qingbai Tongbi capsules was established,identifying 24 common peaks,and assigning them to different peaks.Among them,peaks 1-3,5-8,10-11,13 and 16 came from Qingfengteng.Peaks 4,9,12,14,16,18-19 came from Baishao.Peaks 15,17,20-24 came from Zhigancao.Eight common peaks were identified,including catechin,sinomenine,gallic acid,magnoflorine,paeoniflorin,glycyrrhizin,1,2,3,4,6-0-pentagalloylglucose,glycyrrhizic acid.The similarity evaluation showed that the similarity of 10 batches of Qingbai Tongbi capsules samples ranged from 0.934-1.000.Principal component analysis(PCA)showed that the cumulative variance contribution rate of the first four principal components was 93.998%,while orthogonal partial least squares-discriminant analysis(OPLS-DA)showed that 8 components had higher variable importance projection values.On this basis,the network pharmacology method was used to analyze and conclude that catechin,sinomenine,gallic acid,magnoflorine,paeoniflorin,glycyrrhizin,1,2,3,4,6-0-pentagalloylglucose and glycyrhizic acid may be the potential Q-Marker of Qingbai Tongbi capsules.The contents of the above eight components were determined simultaneously,and the methodological investigation results were good.The average sample recovery rate was 96.81%-103.44%,and the RSD was 0.6%-3.7%.The mass fractions of catechin,sinomenine,gallic acid,magnoflorine,paeoniflorin,glycyrrhizin,1,2,3,4,6-0-pentagalloylglucose and glycyrrhizic acid in 10 batches of samples were 0.9071-1.1893 mg·g^(-1),2.1833-3.1186 mg·g^(-1),0.3970-1.4276 mg·g^(-1),3.5079-5.4466 mg·g^(-1),14.2077-19.5701 mg·g^(-1),1.4128-3.5775 mg·g^(-1),0.4420-1.6977 mg·g^(-1),2.7388-4.7612 mg·g^(-1).Conclusion:The established HPLC fingerprint method is simple and good repeatability.The quality control indicative compounds of Qingbai Tongbi capsules can provide a basis for its quality control.
作者
曲彤
胡筱娟
李宁
鲁文静
耿飞飞
陈音孜
陈志永
任慧
QU Tong;HU Xiao-juan;LI Ning;LU Wen-jing;GENG Fei-fei;CHEN Yin-zi;CHEN Zhi-yong;REN Hui(Shaanxi Academy of Traditional Chinese Medicine,Xian 710003,China;Shaanxi Provincial Hospital of Traditional Chinese Medicine,Xian 710003,China;College of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang 712083,China;School of Life Science,NorthWest University,Xian 710127,China)
出处
《药物分析杂志》
北大核心
2025年第7期1166-1180,共15页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然科学基金面上项目(81973419)
“陕西省中医药管理局双链融合”中青年科研创新团队(2022-SLRH-YQ-003)
陕西省中医药管理局“医研校企”中医药传承创新平台(中医药创新药物/器械“研发-转化-推广”平台)。
