摘要
目的:对坤宁颗粒发挥抗凝血活性的质量标志物(Q-Marker)进行预测分析。方法:采用超高效液相色谱-四极杆-飞行时间串联质谱法(UHPLC-Q-TOF-MS/MS)对坤宁颗粒的化学成分进行分析,检测条件为Waters ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8μm),流动相乙腈(A)-25 mmol·L^(-1)乙酸铵水溶液(B)梯度洗脱(0~5 min,5%~22%A;5~10 min,22%~30%A;10~15 min,30%~95%A;15~20 min,95%~5%A;20~30 min,5%A),流速0.2 mL·min^(-1),柱温30℃,进样量1μL;电喷雾离子源(ESI),正、负离子检测模式。通过网络药理学方法对化学成分的靶点与异常子宫出血(AUB)的靶点进行交互分析,利用网络拓扑分析筛选出关键成分;通过高效液相色谱法(HPLC)建立10批坤宁颗粒的指纹图谱,采用血液凝固法及纤维蛋白原平板法测定坤宁颗粒的抗凝血活性,建立其谱效关系;选择拓扑分析与谱效关系中共有的成分作为Q-Marker,并对其抗凝血活性进行验证和确认。结果:坤宁颗粒含475个化学成分,丹酚酸B、芍药苷、柚皮苷和新橙皮苷等22个关键成分是治疗AUB的潜在物质基础;谱效分析结果表明,峰7(芍药苷)、峰9(柚皮苷)、峰10(新橙皮苷)和峰11(丹酚酸B)为最优主成分,体外活性实验表明这4个成分可较好地表征其抗凝血活性。结论:丹酚酸B、芍药苷、新橙皮苷和柚皮苷可能是坤宁颗粒发挥抗凝血活性的Q-Marker。
Objective:To perform a predictive analysis of the quality marker(Q-Marker)for the anticoagulant activity of Kunning granules.Method:The chemical components of Kunning granules were analyzed by ultra high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry(UHPLC-Q-TOF-MS/MS)on a Waters ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8μm)with the mobile phase of acetonitrile(A)-25 mmol·L^(-1)ammonium acetate aqueous solution(B)for gradient elution(0-5 min,5%-22%A;5-10 min,22%-30%A;10-15 min,30%-95%A;15-20 min,95%-5%A;20-30 min,5%A),flow rate of 0.2 m L·min^(-1),column temperature at 30℃,injection volume of 1μL,electrospray ionization(ESI),positive and negative ion detection modes.Interaction analysis between the targets of chemical components and the targets of abnormal uterine bleeding(AUB)was performed by network pharmacology,and the key components were screened through network topology analysis.The fingerprints of 10 batches of Kunning granules were established by high performance liquid chromatography(HPLC),the anticoagulant activity of the granules was determined by blood coagulation method and fibrinogen plate method,and the spectrum-effective relationship was established.The components co-occurring in the topological analysis and spectrum-effective relationship were selected as Q-Markers,and their anticoagulant activities were verified and confirmed.Result:A total of 475 chemical components were identified from Kunning Granule,of which 22 key components such as salvianolic acid B,paeoniflorin,naringin and neohesperidin,were the potential material basis for the treatment of AUB.The spectrum-effective analysis showed that peaks 7(paeoniflorin),9(naringin),10(neohesperidin)and 11(salvianolic acid B)were the optimal principal components,and in vitro activity test showed that these four components could better characterize their anticoagulant activity.Conclusion:Salvianolic acid B,paeoniflorin,neohesperidin and naringin may be Q-Markers for the anticoagulant activity of Kunning granules.
作者
钟林江
杨放
吴建雄
谢志伟
雷燕莉
田佩灵
贾利军
刘涛
ZHONG Linjiang;YANG Fang;WU Jianxiong;XIE Zhiwei;LEI Yanli;TIAN Peiling;JIA Lijun;LIU Tao(School of Pharmacy,School of Food and Biological Engineering,Sichuan Engineering Research Center of Antiviral Traditional Chinese Medicine Industrialization,Chengdu University,Chengdu 610106,China;Chengdu Tianhe Sino-Western Medical Conservation Co.Ltd.,Chengdu 610000,China;Suzhou Yifan Pharmaceutical Co.Ltd.,Suzhou 234000,China;Sichuan Taiji Pharmaceutical Co.Ltd.,Taiji Group,Chengdu 610207,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第7期168-176,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
四川省2021—2023年高等教育人才培养质量和教学改革项目(JG2021-1106)
四川省科技创新苗子工程项目(2021054)
新药中试四川省重点实验室开放课题(XYZS2022-01)
成都大学2021-2023年研究生人才培养质量和教学改革立项项目(cdjgy2022004)。
