摘要
目的建立大鼠灌胃龙柴方总萜有效部位后血浆中熊果酸、齐墩果酸的HPLC测定方法,并研究其药动学。方法大鼠灌胃龙柴方总萜有效部位(2 g.kg-1)后,不同时间点眼眶后静脉丛取血,制备血浆,血浆样品经甲醇沉淀蛋白,HPLC-UV测定血浆浓度。色谱柱:Kromasil-C18柱(4.6 mm×250 mm,5μm);流动相:甲醇-0.04%磷酸水溶液(88∶12);流速:0.8 mL.min-1;检测波长:220 nm;柱温:35℃;进样量:20μL。熊果酸、齐墩果酸血药浓度-时间数据用药动学软件3P97处理。结果熊果酸和齐墩果酸的线性范围分别为7.40～111.0,7.07～106.05 mg.L-1,定量下限分别为1.04,1.60 mg.L-1,日内、日间精密度(RSD)均小于10%,方法回收率分别为93.5%～100.6%和95.3%～98.6%,提取回收率分别为73.54%～74.36%和73.75%～75.21%。熊果酸、齐墩果酸在大鼠体内的血药浓度-时间过程均符合二房室模型,其中熊果酸Ka=(6.26±1.22)h-1,AUC=(1482.58±18.37)mg.h.L-1,Cmax=(61.54±4.92)mg.L-1,Tmax=(0.50±0.00)h;齐墩果酸Ka=(7.22±1.71)h-1,AUC=(419.40±8.35)mg.h.L-1,Cmax=(49.09±3.92)mg.L-1,Tmax=(0.50±0.00)h。结论该法准确可靠,专属性强,适用于熊果酸及齐墩果酸在大鼠体内的血药浓度测定及药代动力学研究。大鼠灌胃龙柴方总萜有效部位后血浆中熊果酸和齐墩果酸的质量浓度存在双峰现象,熊果酸的2次吸收稍好于齐墩果酸。
Objective To develop a HPLC method with ultraviolet detection for simultaneous determination of ursolic acid and oleanolic acid in rat plasma after intragastrie infusion of total terpenes of Longchai Recipe, and to study the pharmacokineties of ursolic acid and oleanolic acid. Methods The rats were orally treated with 2 g·kg^-1 total tepenes from Longchai Recipe, and then the plasma samples were collected at different time points from the venous plexus of ocular fundus. The plasma concentrations of ursolie acid and oleanolic acid were analyzed by high performance liquid chromatography after protein precipitation with methanol. The analytes were separated on a Kromasil-C18 column （4.6min×250mm,5μm） using methanol-0.04 % phosphoric acid （40 ： 60, V/V） as a mobile phase at a flow rate of 0.8 mL＇min-1. The detection wavelength was 220 nm, the column temperature was 35 ℃, Sample volume was 20 μL. The pharmacokinetic parameters were calculated by the software of 3P97. Results A good linear relationship of ursolic acid was obtained in the range of 7.40 to 111.0mg·L^-1, and the limit of quantitation was 1.04 mg·L^-1.A good linear relationship of oleanolic acid was obtained in the range of 7.07 to 106.05 mg·L^-1, and the limit of quantitation was 1.60 mg·L^-1. The intra-day and inter-day precision of this method was both less than 10 %. For ursolic acid and oleanolic acid, the method recoveries ranged from 93.5 % to 100.6 %, 95.3 % to 98.6 %, and the extraction recoveries ranged from 73.54 % to 74.36 %, 73.75 % to75.21%, respectively. The plasma ursolic acid and oleanolic acid coneentra tion-time curve could be evaluated by two-compartment model. The main pharmacokinetic parameters of Ka, AUC, Cmax, Tmax were 6.26 ± 1.22 h^-1, 1482.58 ± 18.37 mg·h·L^-1, 61.54 ± 4.92 mg·L^-1, 0.50 ± 0.00 h for the ursolic acid, and were 7.22 ± 1.71 h^-1, 419.40 ± 8.35 mg·h·L^-1, 49.09 ± 3.92 mg·L^-1, 0.50 ± 0.00 h for oleanolic acid. Conclusion The method has been successfully applied to the determination of ursolic acid and oleanolic acid in rat plasma. Plasma concentration-time curves of ursolic acid and oleanolic acid show the existence of double peaks after intragastric admin-istration of total terpenes of Lonchai Recipe. The second absorption of ursolic acid is better than that of oleanolic acid.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2013年第3期267-271,共5页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金(810729851H2801)
江苏省"科技基础实施建设计划"专项资助项目(BM2009903
BM2009903-KF22)
2010年江苏省高等学校大学生实践创新训练计划立项项目(529)
关键词
龙柴方
总萜
熊果酸
齐墩果酸
高效液相色谱法
药物动力学
Lonchai Recipe
Total terpene
Ursolic acid
Oleanolic acid
High pressure liquid chromatography
Pharmacokinetics
