摘要
目的:建立HPLC测定绿茶浸出液中没食子酸、咖啡因、表儿茶素没食子酸酯、山柰酚在动脉粥样硬化大鼠体内的血药浓度,为动脉粥样硬化的治疗提供参考。方法:动脉粥样硬化大鼠灌胃绿茶浸出液后,分别在不同时间点取血,采用依利特Hypersil ODS2色谱柱(4.6 mm×250 mm,5μm),流动相甲醇(A)-0.03%磷酸水溶液(B)梯度洗脱(0~30 min,10%~43%A;30~43 min,43%~45%A;43~44 min,45%~51%A;44~60 min,51%~53%A;60~70 min,53%~90%A),流速0.7 mL·min^(-1),检测波长278 nm的色谱条件进样分析。结果:没食子酸、咖啡因、表儿茶素没食子酸酯、山柰酚分别在0.255~20.40,0.430~86.00,0.245~19.60,0.270~21.60 mg·L^(-1)内呈良好线性关系,日内和日间精密度RSD均<9.0%,提取回收率和方法回收率都>87%;酸化后主要药代动力学参数药时曲线下面积(AUC0-t)分别为(8.902±1.235),(198.5±12.5),(7.367±0.412),(4.426±0.251)mg·h·L^(-1),表观分布容积(Vd)分别为(0.990±0.134),(1.409±0.201),(0.501±0.243),(0.029±0.009)L·kg^(-1),达峰时间(T_(max))均为1.5 h。结论:该方法对所测成分的分离度好,适用于测定绿茶浸出液中4个有效成分的血药浓度,为临床预防此类疾病提供一定理论依据。
Objective： To establish a HPLC method for determination of gallic acid, caffeine, epicatechin gallate and kaempferol in plasma of rats with atherosclerosis after intragastric administration of green tea extract. Method： After being administrated green tea extract, the rats＇ blood were taken at different time points, separation was carried out on a Hypersil ODS2 column （4.6 mm×250 mm, 5 μm） , the mobile phase was methanol-0.03% phosphoric acid aqueous solution, the flow rate was 0.7 mL·min^-1 and the detection wavelength was set at 278 nm, the column temperature was maintained at 30℃. Result： The calibration curves of gallic acid, caffeine, epicatechin gallate and kaempferol were in good linearity over the range of 0. 255-20.40, 0. 430-86.00,0.245-19.60, 0.270-21.60 mg·L^-1. RSDs of inter-day and intra-day assay were less than 9.0%. The average method separating degree for four ingredients and their extraction recoveries were more than 87%. After acidification, main pharmacokinetic parameters of the area under the curve （ AUC0-t） were （8. 902±1. 235 ） , （ 198.5 ±12.5 ）, （7. 367 ± 0. 412）, （4. 426± 0.251 ） mg·h·L^-1 and the apparent volume of distribution （ Vd ） were （0.990±0.134）, （1.409±0.201）, （0.501±0.243）, （0.029±0.009） L·kg^-1, respectively; the peak time （ Tmax） of these four ingredients were 1.5 h. Conclusion： This method has good separation which can be used to detect blood drug concentration of four effective ingredients from green tea extract.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第8期92-97,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金地区科学基金项目(81560720)
喀斯特地区生物与信息技术协同创新中心项目(黔教合协同创新字[2014]04)
