姜丽;余兰彬;徐国良;张启云;严小军;王玉蓉;
JIANG Li;YU Lan-bin;XU Guo-liang;ZHANG Qi-yun;YAN Xiao-jun;WANG Yu-rong;Research Center for Differentiation and Development of Basic Traditional Chinese Medicine Theory,Jiangxi University of Traditional Chinese Medicine;School of Chinese Materia Medica,Beijing University of Chinese Medicine;

• 1:江西中医药大学中医基础理论分化发展研究中心
• 2:北京中医药大学中药学院

摘要(Abstract)：

建立同时测定天麻素及其代谢产物天麻苷元、葛根素和内标对羟基苯乙醇(Tyr)4种成分在大鼠血浆中药物浓度的HPLC方法,并应用该法研究单独或合并静脉注射及灌胃给药天麻素、葛根素后,三者在大鼠体内药代动力学过程及相互影响。以Tyr为内标,血浆样品经甲醇沉淀蛋白后,上清液用N2吹干,残渣用乙腈-0.05%磷酸(20∶80)复溶后,以乙腈-0.05%磷酸水为流动相梯度洗脱,用Agilent ZORBAX SB-Aq C18(4.6 mm×150 mm,5μm)色谱柱分离,在221 nm处测定天麻素、天麻苷元和内标,250 nm处测定葛根素。单独或合并给药天麻素、葛根素后,检测大鼠血浆中三者浓度,用Win Nonlin 5.2药动学软件和SPSS 17.0软件分别计算各给药组药动学参数并进行统计分析,比较单独或合并给药后天麻素、葛根素药动学过程。天麻素、天麻苷元和葛根素低、中、高浓度及内标对羟基苯乙醇的绝对回收率均>77.20%;天麻素、天麻苷元、葛根素分别在0.10~101,0.03~7.58,0.05~5.98 mg·L-1具有良好的线性(r>0.999 6,n=5),三者定量下限分别为0.10,0.03,0.05 mg·L-1,日内及日间精密度均<12%,准确度85.61%~110.3%,且测定物在整个分析过程中稳定。结果表明,天麻素和葛根素联合给药后,大鼠体内主要药代动力学参数(AUC,Cmax,T1/2,Tmax,MRT)与单独给药后有显著性差异。无论是灌胃还是静脉注射,清除率(CL)降低,平均滞留时间(MRT)延长;相对生物利用度均有提高,尤以灌胃组效果显著,其中灌胃天麻素合用组为单用组的1.5倍,灌胃葛根素合用组为单用组的10.7倍。天麻素和葛根素合用后能相互促进吸收,降低消除速率,增加体内滞留时间。该法可同时测定大鼠体内天麻素、天麻苷元和葛根素血药浓度和药动学,且方法准确,操作简便。
To establish a HPLC method for simultaneously determining plasma concentrations of gastrodin( Gas) and its metabolites hydroxybenzyl alcohol( HBA),puerarin( Pur) and internal standard( IS) p-hydroxyphenylethanol( Tyr) in rats and studying the pharmacokinetic process and interactions of gastrodin and puerarin after single and combined intravenous injection and oral administration. With Tyr as the internal standard,plasma samples were processed with methanol for protein precipitation,supernatant was dried with N2,and residues were re-dissolved with acetonitrile-0. 05% phosphoric acid( 20∶ 80). Chromatography was carried out on an Agilent ZORBAX SB-Aq C18column( 4. 6 mm × 250 mm,5 μm),with acetonitrile-0. 05% phosphoric acid as the gradient mobile phase for the gradient elution. The UV detector wavelength was set at 221 nm for Gas HBA and IS and 250 nm for Pur. After the single or combined administration of Gas and Pur,their plasma concentrations in rats were detected. Win Nonlin 5. 2 pharmacokinetic software and SPSS 17. 0 software were used to respectively calculate pharmacokinetic parameters of each group,make a statistical analysis and compare the pharmacokinetic processes of Gas and Pur after the single or combined administration. According to the results,the absolute recoveries between low,media and high concentrations of Gas,HBA and Pur and IS as well as Tyr were more than 77. 20%,with a good linearity( r > 0. 999 6,n = 5) for Gas,HBA and Pur within concentration ranges of 0. 10-101,0. 03-7. 58 and 0. 05-5. 98 mg·L-1,respectively. The lower limits of quantification for Gas,HBA and Pur were 0. 10,0. 03,0. 05 mg·L-1,respectively. Their intra-day and inter-day precisions were less than 12%,with the accuracy between 85. 61%-110. 3%. All of the three substances and IS were stable during the whole analysis process. The findings showed significant differences in the main in vivo pharmacokinetic parameters in rats( AUC,Cmax,T1 /2,Tmax,MRT) after the single and combined administration of Gas and Pur. Either after the oral administration or after the intravenous injection,parameters showed a lower clearance rate( CL),longer mean residence time( MRT) and higher relative bioavailability,especially after the oral administration. Specifically,the relative bioavailability of the combined oral administration of Pur was 10. 7 times of that of the single administration,while that of Gas was 1. 5 times of that of the single administration. The combined administration of Gas and Pur can promote the absorption,decrease the elimination rate and prolong the mean residence time. The method is simple and accurate and can be applied in the simultaneous determination of plasma concentrations of Gas,HBA and Pur in rats and the pharmacokinetic studies.

关键词(KeyWords)： 天麻素;天麻苷元(对羟基苯甲醇);对羟基苯乙醇;葛根素;RP-HPLC;药代动力学;大鼠血浆
gastrodin;gastrodigenin(hydroxybenzyl alcohol);tyrosol;puerarin;RP-HPLC;pharmacokinetics;rat plasma

Abstract:

Keywords:

基金项目(Foundation): 国家重点基础研究发展计划(973)项目(2010CB530603)

作者(Author): 姜丽;余兰彬;徐国良;张启云;严小军;王玉蓉;
JIANG Li;YU Lan-bin;XU Guo-liang;ZHANG Qi-yun;YAN Xiao-jun;WANG Yu-rong;Research Center for Differentiation and Development of Basic Traditional Chinese Medicine Theory,Jiangxi University of Traditional Chinese Medicine;School of Chinese Materia Medica,Beijing University of Chinese Medicine;

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DOI:

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