夏东亚郭涛潘文灏的等hplc法测定心舒口服液中阿魏酸血浆.doc

高效液相色谱法测定比格犬血浆中阿魏酸浓度及其药代动力学研究 黄莉莉2，北京，2.南京大学医学院附属鼓楼医院，南京, 210008) 摘要 目的：建立高效液相色谱法测定比格犬血浆中阿魏酸的含量，并用于药代动力学研究。方法：血浆样品采用乙酸乙酯处理，以替硝唑作为内标，色谱柱为Phenomenex Luna C18色谱柱(150 mm×4.6 mm, 5μm)，流动相为水(含5 mmol?L-1醋酸铵和0.05%H3PO4)-乙腈(60︰40)，等度洗脱，流速为1.0 mL·min-1，紫外检测波长为320 nm。采用比格犬单剂量静注3、6、12 mg·kg-1的阿魏酸，HPLC-UV法测定阿魏酸的血药浓度，并采用DAS 2.0软件计算药代动力学参数。结果：方法学实验结果表明内源性杂质不干扰阿魏酸和内标的测定，线性范围0.0975～12.5 μg·mL-1，定量限为0.0975 μg·mL-1。方法精密度、准确度、稳定性和回收率均符合生物样品测定的要求，适合比格犬血浆中阿魏酸浓度的测定，可以应用该方法进行阿魏酸的药代动力学研究。比格犬单剂量静注3、6、12 mg·kg-1的阿魏酸后，血药浓度-时间曲线下面积(AUC0-τ)分别为244.81±57.34、508.40±118.82、988.87±172.29 min?μg·mL-1，不同剂量下AUC0-τ比为1:2.1:4.0，与剂量比1:2:4近似成比例，说明在研究的剂量范围内，阿魏酸在Beagle犬体内的消除过程是线性的。结论：本方法操作简便、灵敏、专属性强，方法学考证符合生物样品测定的要求并成功用于阿魏酸在比格犬体内的药代动力学研究。 关键字：阿魏酸；高效液相色谱；药代动力学；比格犬血浆 中图分类号R28；R97 文献标志码 ADevelopment and Validation of an HPLC-UV Method for Determination of Ferulic Acid in Beagle Dog Plasma and Its Application to a Pharmacokinetic Study Xu Sun1, Huang Li-li2, Wang Xin 2*（1. National Center for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, Beijing 100050，China；2. Pharmaceutical Preparation Section of Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, Nanjing 210008，China） Abstract Objective: To establish a sensitive, simple and specific high performance liquid chromatography method for determination of ferulic acid in order to study the pharmacokinetics of ferulic acid in beagle dogs. Methods: Ferulic acid and tinidazole (internal standard, IS) were extracted by liquid–liquid extraction and separated on a Phenomenex Luna C18 column, with acetonitrile and water (containing 5 mmol?L-1 ammonium acetate and 0.05% phosphoric acid) (40:60) as the mobile phase. The flow rate was set to be 1.0 mL/min and the analytical wavelength was 320 nm. After validating, the developed method was used for evaluating pharmacokinetics of ferulic acid in beagle dogs following intravenous administration of three doses (3, 6, 12 mg/kg) of ferulic acid. The pharmacokinetic