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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 the determination of plasma concentrations of gallic acid, caffeine, epicatechin gallate and kaempferol in atherosclerotic rats in green tea extract, and provide a reference for the treatment of atherosclerosis. Methods: After the rats were instilled with green tea leachate, the rats were sacrificed at different time points. Eelite Hypersil ODS2 column (4.6 mm × 250 mm, 5 μm), mobile phase of methanol (A) -0.03% phosphoric acid solution (B) gradient elution (0-30 min, 10-43% A; 30-43 min, 43-45% A; 43-44 min, 45-51%; 44-60 min, 51% ~ 53% A, 60 ~ 70 min, 53% ~ 90% A), the flow rate was 0.7 mL · min ~ (-1) and the detection wavelength was 278 nm. Results: There was a good linear relationship between gallic acid, caffeine, epicatechin gallate and kaempferol at 0.255 ~ 20.40, 0.430 ~ 86.00, 0.245 ~ 19.60, 0.270 ~ 21.60 mg · L -1, The intra-day and inter-day RSDs were less than 9.0%, and the recovery rates and recovery rates were both> 87%. The mean pharmacokinetic parameters (AUC0-t) after acidification were 8.902 ± 1.235, (198.5 ± 12.5), (7.367 ± 0.412) and (4.426 ± 0.251) mg · h · L -1, and the apparent volume of distribution (Vd) were 0.990 ± 0.134 and 1.409 ± 0.201, respectively. 0.501 ± 0.243), (0.029 ± 0.009) L · kg -1, peak time (T max) were both 1.5 h. Conclusion: The method is good for the separation of the tested components and is suitable for determining the plasma concentration of four active ingredients in the green tea extract, providing a theoretical basis for the clinical prevention of such diseases.