LC-MS/MS同时测定大鼠血浆中瑞香素、西瑞香素和瑞香新素及其药代动力学研究Simultaneous determination of daphnetin,daphnoretin,daphneticin in rat plasma by LC-MS/MS and its application in pharmacokinetic study
胡伊力格其;曹萨丽;林龙飞;付京;董晓旭;杨春静;张淼;倪健;
HU Yi-yi-li-ge-qi;CAO Sa-li;LIN Long-fei;FU Jing;DONG Xiao-xu;YANG Chun-jing;ZHANG Miao;NI Jian;College of Traditional Mongolia Medicine and pharmacy,Inner Mongolia University for Nationalities;Beijing University of Chinese Medicine;
摘要(Abstract):
建立同时测定大鼠灌胃祖师麻提取物后血浆中瑞香素、西瑞香素和瑞香新素的HPLC-MS/MS分析方法,并用于计算3个成分在大鼠体内的药代动力学参数。试验采用6只SD大鼠,口服给予祖师麻提取物(瑞香素、西瑞香素和瑞香新素的给药量分别是88.40,3.24,4.28 mg·kg(-1)),以五味子醇甲为内标,用LC-MS/MS测定给药后血浆中药物浓度,绘制血药浓度-时间曲线,并用Kinetica 4.4软件计算相关药代动力学参数。试验结果显示,瑞香素、西瑞香素和瑞香新素的线性范围均为5(-1)),以五味子醇甲为内标,用LC-MS/MS测定给药后血浆中药物浓度,绘制血药浓度-时间曲线,并用Kinetica 4.4软件计算相关药代动力学参数。试验结果显示,瑞香素、西瑞香素和瑞香新素的线性范围均为51 000μg·L1 000μg·L(-1),方法学考察符合要求。日内、日间变异系数(RSD)均小于15%,精密度和准确度符合生物样品分析要求。瑞香素、西瑞香素和瑞香新素口服给药后的T_(max)分别为4,2.92,2 h;C_(max)分别为858.96,178.00,36.67μg·L(-1),方法学考察符合要求。日内、日间变异系数(RSD)均小于15%,精密度和准确度符合生物样品分析要求。瑞香素、西瑞香素和瑞香新素口服给药后的T_(max)分别为4,2.92,2 h;C_(max)分别为858.96,178.00,36.67μg·L(-1);AUC_(0-t)分别为10 566.4,905.89,355.11μg·L(-1);AUC_(0-t)分别为10 566.4,905.89,355.11μg·L(-1)·h;半衰期t1/2分别为5.19,3.50,4.95 h;平均驻留时间MRT分别为9.43,6.95,8.27 h。因此,该研究建立的LC-MS/MS分析方法准确灵敏,适于对SD大鼠口服祖师麻提取物后的瑞香素、西瑞香素和瑞香新素的药代动力学研究。
To establish HPLC-MS/MS method for simultaneous determination of daphnetin,daphnoretin,and daphneticin in rat plasma after oral and intravenous administration of Daphne giraldii extract,and then use them in the calculation of pharmacokinetic parameters. Six sprague-dawley rats received intragastric administration of D. giraldii extract(daphnetin,daphnoretin and daphneticin were 88. 40,3. 24 and 4. 28 mg·kg,respectively). Their drug plasma concentration was determined by LC-MS/MS with schisandrin as an internal standard to draw plasma concentration-time curve. The pharmacokinetic parameters were calculated by Kinetica 4. 4. The results showed that the linear range was 5-1 000 μg·L(-1)·h;半衰期t1/2分别为5.19,3.50,4.95 h;平均驻留时间MRT分别为9.43,6.95,8.27 h。因此,该研究建立的LC-MS/MS分析方法准确灵敏,适于对SD大鼠口服祖师麻提取物后的瑞香素、西瑞香素和瑞香新素的药代动力学研究。
To establish HPLC-MS/MS method for simultaneous determination of daphnetin,daphnoretin,and daphneticin in rat plasma after oral and intravenous administration of Daphne giraldii extract,and then use them in the calculation of pharmacokinetic parameters. Six sprague-dawley rats received intragastric administration of D. giraldii extract(daphnetin,daphnoretin and daphneticin were 88. 40,3. 24 and 4. 28 mg·kg,respectively). Their drug plasma concentration was determined by LC-MS/MS with schisandrin as an internal standard to draw plasma concentration-time curve. The pharmacokinetic parameters were calculated by Kinetica 4. 4. The results showed that the linear range was 5-1 000 μg·L(-1) for daphnetin,daphnoretin and daphneticin,and the method ological test showed conformance to the requirements. The intraday and inter-day variable coefficients(RSD) were both less than 15. 0%,indicating that both of legitimate precise and accuracy were consistent with the analysis requirements of biological samples. For daphnetin,the pharmacokinetic parameters Tmax,Cmax,AUC0–t,T_(1/2) and MRT were 4 h,858. 96 μg·L(-1) for daphnetin,daphnoretin and daphneticin,and the method ological test showed conformance to the requirements. The intraday and inter-day variable coefficients(RSD) were both less than 15. 0%,indicating that both of legitimate precise and accuracy were consistent with the analysis requirements of biological samples. For daphnetin,the pharmacokinetic parameters Tmax,Cmax,AUC0–t,T_(1/2) and MRT were 4 h,858. 96 μg·L(-1),10 566. 4 μg·L(-1),10 566. 4 μg·L(-1)·h,5. 19 h and9. 43 h,respectively. For daphnoretin,the pharmacokinetic parameters T_(max),C_(max),AUC_(0–t),T_(1/2) and MRT were 2. 92 h,178. 00 μg· L(-1)·h,5. 19 h and9. 43 h,respectively. For daphnoretin,the pharmacokinetic parameters T_(max),C_(max),AUC_(0–t),T_(1/2) and MRT were 2. 92 h,178. 00 μg· L(-1),905. 89 μg· L(-1),905. 89 μg· L(-1)· h,3. 50 h and 6. 95 h,respectively. For daphneticin,the pharmacokinetic parameters T_(max),C_(max),A UC_(0–t),T_(1/2) and MRT were 2 h,36. 67 μg·L(-1)· h,3. 50 h and 6. 95 h,respectively. For daphneticin,the pharmacokinetic parameters T_(max),C_(max),A UC_(0–t),T_(1/2) and MRT were 2 h,36. 67 μg·L(-1),355. 11 μg·L(-1),355. 11 μg·L(-1)·h,4. 95 h and 8. 27 h,respectively. The LC-MS/MS analysis method established in this study was proved to be so accurate and sensitive that it can be applied to the pharmacokinetic study of daphnetin,daphnoretin and daphneticin.
关键词(KeyWords):
LC-MS/MS;祖师麻;瑞香素;西瑞香素;瑞香新素;药代动力学
LC-MS/MS;Daphne giraldii;daphnetin;daphnoretin;daphneticin;pharmacokinetic
基金项目(Foundation):
作者(Authors):
胡伊力格其;曹萨丽;林龙飞;付京;董晓旭;杨春静;张淼;倪健;
HU Yi-yi-li-ge-qi;CAO Sa-li;LIN Long-fei;FU Jing;DONG Xiao-xu;YANG Chun-jing;ZHANG Miao;NI Jian;College of Traditional Mongolia Medicine and pharmacy,Inner Mongolia University for Nationalities;Beijing University of Chinese Medicine;
DOI: 10.19540/j.cnki.cjcmm.20170307.003
参考文献(References):
- [1]中国药典.一部[S].1977:445.
- [2]苏娟.黄瑞香化学成分研究[D].上海:第二军医大学,2007.
- [3]叶和杨,熊小琴,邱伟,等.瑞香素对醋酸、热板及电刺激致痛小鼠的镇痛作用[J].中国临床康复,2005,22(9):174.
- [4]杨鸽华,王丽丽,封玲,等.祖师麻总香豆素的抗炎镇痛药效作用评价[J].中华中医药学刊,2015,33(5):1183.
- [5]张立,喻文进,刘慧琼,等.西瑞香素抗炎抑菌作用的初步实验研究[J].中医药导报,2012,18(6):72.
- [6]王佳,黄俊卿.瑞香素对小鼠胶原诱导性关节炎的抑制作用[J].亚太传统医药,2015,11(19):13.
- [7]涂苑青,黄丰,朱苗苗,等.瑞香素对脂多糖诱导的小鼠小胶质细胞炎症反应的保护作用[J].时珍国医国药,2013,24(5):1169.
- [8]戚之琳,齐世美,凌烈锋,等.瑞香素对HMGB1释放及HMGB1诱发的炎症反应的双重抑制作用[J].南方医科大学学报,2015,35(11):1519.
- [9]姜洪芳,白雪,牛慧彦,等.西瑞香素对肺癌A549细胞侵袭及迁移能力的影响[J].实用药物与临床,2016,19(2):131.
- [10]况南珍,傅颖媛.瑞香素对SMMC-7721细胞凋亡及Bcl-2/Bax基因mRNA表达的影响[J].广东医学,2012,33(10):1374.
- [11]杨振宇,郭薇,吴东媛,等.了哥王中西瑞香素的提取分离及抗肿瘤作用研究[J].天然产物研究与开发,2008,20(3):522.
- [12]刘嘉,狄留庆,单进军,等.瑞香素不同给药途径在大鼠体内药动学研究[J].中草药,2009,40(1):106.
- [13]陈乐天,狄留庆,刘卉,等.甘草提取物对瑞香素大鼠体内药代动力学特征的影响研究[J].中国中药杂志,2011,36(7):935.
- [14]Zhang W,Di L,Li J S,et al.The effects of Glycyrrhiza uralenis and its major bioactive components on pharmacokinetics of daphnetin in Cortex daphnes in rats[J].J Ethnopharmacol,2014,154(3):584.
- [15]Lin L C,Yang K Y,Chen Y F,et al.Measurement of daphnoretin in plasma of freely moving rat by liquid chromatography[J].J Chromatogr A,2005,1073(1):285.
- [16]Sun L X,Chen Y,Yang W W,et al.Development and validation of an RP-HPLC-UV method for the determination of daphnoretin in rat plasma[J].J Chin Pharm Sci,2011,20:493.
- [17]郑玲,高金薇,张石,等.西瑞香素在大鼠体内组织分布研究[J].中国药学杂志,2013,48(11):884.
- 胡伊力格其
- 曹萨丽
- 林龙飞
- 付京
- 董晓旭
- 杨春静
- 张淼
- 倪健
HU Yi-yi-li-ge-qi- CAO Sa-li
- LIN Long-fei
- FU Jing
- DONG Xiao-xu
- YANG Chun-jing
- ZHANG Miao
- NI Jian
- College of Traditional Mongolia Medicine and pharmacy
- Inner Mongolia University for Nationalities
- Beijing University of Chinese Medicine
- 胡伊力格其
- 曹萨丽
- 林龙飞
- 付京
- 董晓旭
- 杨春静
- 张淼
- 倪健
HU Yi-yi-li-ge-qi- CAO Sa-li
- LIN Long-fei
- FU Jing
- DONG Xiao-xu
- YANG Chun-jing
- ZHANG Miao
- NI Jian
- College of Traditional Mongolia Medicine and pharmacy
- Inner Mongolia University for Nationalities
- Beijing University of Chinese Medicine