【作者】 刘晓峰；

【导师】 武新安；

【作者基本信息】 兰州大学 ， 药物分析学， 2007， 硕士

【摘要】 目的：建立简单、灵敏、重现性好的高效液相色谱方法，对川芎活性成分阿魏酸的血浆浓度进行测定，并将其应用于药动学研究中，探讨阿魏酸单体与川芎煎剂中阿魏酸的药动学是否相同以及三种不同溶媒（水、乙醇和乙酸乙酯）的川芎提取物中阿魏酸的药动学是否相同。方法：采用Cosmosil-C₁₈分析柱，流动相为甲醇-3％醋酸（32：68，v／v），流速0.8 mL／min，于323 nm波长处检测，柱温为室温，进样量20μL。大鼠分别灌胃阿魏酸单体溶液、川芎煎剂和三种不同溶媒川芎提取物的溶液或通过动脉插管瞬间注射阿魏酸溶液后，于大鼠股动脉采集血样。用RP-HPLC法测定血浆中阿魏酸的浓度，并用DAS1.0软件估算相应的药动学参数。结果：（1）阿魏酸和内标的保留时间分别为7.7 min和12.5 min，色谱峰尖锐对称，无血浆内源性物质干扰。阿魏酸在高、低浓度范围（5.0-50.0μg／mL，0.1-5.0μg／mL）线性为y=0.7139x-0.3738和y=0.6517x+0.0066，其相关系数r分别为0.9999和0.9994，日内日间准确度均高于91.95％、精密度RSD均小于7.76％，回收率达到87.93％。信噪比为10时，阿魏酸的最低定量限为0.03μg／mL。（2）在相同的阿魏酸给药剂量（30 mg／kg）下，大鼠分别灌胃给予阿魏酸溶液和川芎煎剂，阿魏酸单体的T_max为9min，其平均C_max为32.67μg／mL，而川芎煎剂中阿魏酸的T_max仅为2.4 min，其平均C_max为2.29μg／mL。三种不同溶媒的川芎提取物大鼠灌胃后阿魏酸的t_1／2a，t_1／2β，Vd／F，CL／F，AUC_0-t，T_max，C_max存在一定差别，其中乙醇提取物大鼠灌胃后阿魏酸的AUC_0-t为（2.89±0.44）mg／L^*h，而水提取物和乙酸乙酯提取物大鼠灌胃后其AUC_0-t分别为（1.27±0.55）mg／L^*h和（1.43±0.44）mg／L^*h，水提取物，乙醇提取物，乙酸乙酯提取物大鼠灌胃后阿魏酸的t_1／2a分别为1.62±1.21 h、0.58±1.19 h和0.42±0.97 h，t_1／2β分别为6.44±0.50h、7.02±0.36 h和3.21±0.17 h。结论：（1）阿魏酸和川芎煎剂中阿魏酸在等量给药剂量下给大鼠灌胃，其C_max相差约14倍，T_max时间相差约5倍，药动学参数t_1／2a，t_1／2β，AUC_0-∞存在显著性差异。因此，不能简单地用某种单体的药动学参数来推测单味中药或复方中药有效成分的体内过程及其药动学参数。（2）经统计学分析，三种不同溶媒川芎提取物中阿魏酸在大鼠体内的药动学参数t_1／2a，t_1／2β，AUC_0-t存在显著性差异。其中乙醇提取物中阿魏酸大鼠灌胃后的AUC_0-t最大，生物利用度最高，t_1／2β最长。水提取物中阿魏酸大鼠灌胃后的AUC_0-t最小，生物利用度最低，t_1／2a最长。由此可见在中药的提取过程中，应该注重溶媒的选择，否则可能会影响其活性成分的体内过程。创新性：（1）首次考察了川芎不同溶媒提取物中阿魏酸的药动学。（2）首次采用反卷积法对阿魏酸在大鼠体内的吸收进行了估算。更多还原

【Abstract】 Objective: The purpose of the present work is to develop a simple, sensitive and specific HPLC assay for the quantitation of active components of traditional Chinese medicine in rat plasma and its application in pharmacokinetics study, and to investigate whether the pharmacokinetics of ferulic acid and ferulic acid in Chuanxiong extraction of different solutions were similary or not.Methods: The separation was performed on a Cosmosil-C₁₈ column. The mobile phase consisted of methanol-3% acetic acid solution in a volume 32:68 （v:v）. The mobile phase was delivered at a flow rate of 0.8 mL/min. Detection was performed at a wavelength of 323 nm at room temperature. The sample injection volume was 20μL. The HPLC was used to determine the plasma concentration of ferulic acid after oral administration or bolus injecting ferulic acid solution in rat plasma. Moreover, their pharmacokinetics parameters were estimated by DAS 1.0 software.Results: （1） The retention time for internal standard and ferulic acid were approximately 7.7 min and 12.5 min, respectively The linear regression analysis of the standard calibration plot for rat plasma were y = 0.7139 x-0.3738 for the high concentration range （5.0-50.0μg/mL） and y = 0.6517 x + 0.0066 for the low concentration （0.1-5.0μg/mL） with r values of 0.9999 and 0.9994, respectively. The RSD of ferulic acid was lower than 7.76% for intra-day and inter-day assays. Assay accuracy was higher than 91.95%. The average recovery of ferulic acid was more than 87.93%. The limit of quantitation was deemed to be 0.03μg/mL.（2） The mean maximum concentration of ferulic acid and ferulic acid in Chuanxiong decoction in plasma was 32.67 and 2.29μg/mL at 9.0 and 2.4 min after oral administration with a dose of 30 mg/kg or equivalent to 30 mg/kg ferulic acid of Chuanxiong extraction, respectively. The t_1/2a, t_1/2β, Vd/F, CL/F, AUC_0-t, T_max and C_max have significantly differences following oral administration of Chuanxiong extraction of different solutions. The AUC_0-t were 1.27±0.55 mg/L*h, 1.43±0.44 mg/L*h and 2.89±0.44 mg/L*h after oral administration of aqueous solution, ethyl acetate and alcoholic extraction. respectively. The t_1/2a and t_1/2β were 1.62±1.21 h, 0 .58±1.19 h, 0.42±0.97 h and 6.44±0.50 h, 7.02±0.36 h, 3.21±0.17 h after oral administration of aqueous solution, alcoholic and ethyl acetate extraction, respectively.Conclusions: （1） The maximum concentration and time of maximum concentration of oral administration ferulic acid and ferulic acid in Chuanxiong decoction had significant differences. The maximum concentration of ferulic acid in plasma was more 14 times than that in Chuanxiong decoction, but the peak time of maximum concentration of ferulic acid was less 5 times than that in Chuanxiong decoction. The results suggested that the absorption of ferulic acid after oral administration of ferulic acid in Chuanxiong decoction was much lower than ferulic acid in rat. Therefore, the pharmacokinetics parameters of monomer following oral administration cannot simply presume those of active component of traditional Chinese medicine.（2） With statistics analysis, the ferulic acid of pharmacokinetics parameter t_1/2a, t_1/2β, AUC_0-t had significant differences in Chuanxiong extraction with different solutions in rat vivo. The AUC_0-t and bioavailability of alcoholic extraction was larger than those of aqueous solution extraction or ethyl acetate extraction. The t_1/2a aqueous solution extraction and t_1/2β of alcoholic extraction were slow. The choice of different solutions should be noticed in extraction process of traditional Chinese medicine, and avoided influencing the physiological disposition of active components.New ideas: （1） Pharmacokinetics of ferulic acid of Chuanxiong extracted with different solutions was investigated for the first time. （2） The absorption of ferulic acid was estimated in rat vivo by deconvolution methods for the first time.更多还原