高效液相色谱法测定凉血通瘀方给药大鼠血清中大黄酸的浓度  

HPLC Method for the Determination of Rhein in Rat Serum after Oral Administration of Blood-cooling and Stasis-removing Prescription

作　　者：傅静娟[1] 邓海山[2] 赵凤鸣[3] 吴勉华[3] 谷晶[3] 

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]南京中医药大学药学院,江苏南京210023 [3]南京中医药大学基础医学院,江苏南京210023

出　　处：《现代中药研究与实践》2016年第2期28-30,共3页Research and Practice on Chinese Medicines

基　　金：国家科技部973计划基金资助项目(2006CB504807);江苏省"青蓝工程"优秀青年骨干教师资助项目(2012)

摘　　要：目的测定大鼠灌服凉血通瘀方后血清中大黄酸的浓度。方法采用HPLC法测定大鼠含药血清中大黄酸的浓度,血清样品经H_2SO_4溶液酸化后,用乙酸乙酯提取,色谱柱为XTerra RP18,流动相为甲醇-0.1%磷酸（85∶15,v/v）,检测波长为431 nm。结果大黄酸在血清浓度0.04~8μg/ml范围内具有良好的线性关系,日内精密度为2.6%~5.2%,日间精密度为2.1%~6.8%,方法回收率为96.6%~98.2%,提取回收率为87.5%~89.2%。0.5、1、2、4 g/ml用药组的含药血清中大黄酸浓度分别为0.12、0.38、0.3、0.39μg/ml。结论建立的大鼠含药血清中大黄酸浓度的测定方法准确、精密、灵敏,大鼠灌服凉血通瘀方后血清中大黄酸浓度的测定结果可为作用机制研究及应用提供理论和实验依据。Objective To determine the concentration of rhein in rat serum after oral administration of Blood- cooling and Stasis-removing prescription. Methods The concentration of rhein in rat serum was determined by HPLC. The sample was acidified by adding sulfuric acid solution, and then extracted by ethyl acetate. XTerra RP18 column was used to separate the analyte. The mobile phase was consisted of methanol and 0.1% phosphoric acid （85 ： 15, v/v）, and the detection wavelength was 431 nm. Results Good linearity was maintained within the concentration range of 0.04 ~tg/ml to 8 ktg/ml. The intra-day precisions were between 2.6% and 5.2%. The interday precisions were between 2.1% and 6.8%. The method recoveries were between 96.6% and 98.2%, and the extraction recoveries were between 87.5% and 89.2%. The concentrations ofrhein in rat serum were 0.12, 0.38, 0.34 μg/ml and 0.39 μg/ml for 0.5, 1, 2 and 4 g/ml dosage groups, respectively. Conclusion The method is accurate, precise and sensitive. The determined concentrations of rhein in rat serum after oral administration of Blood- cooling and Stasis-removing prescription will provide theoretical and experimental foundation for the research of mechanism and the application of the prescription.

关 键 词：凉血通瘀方 大黄酸 含药血清 高效液相色谱法 

分 类 号：R285.5[医药卫生—中药学]