文爱东, 黄熙, 蒋永培, 樊亚萱. HPLC法测定血瘀证兔血清中游离阿魏酸J. 药学学报, 1995, 30(10): 762-767.
引用本文: 文爱东, 黄熙, 蒋永培, 樊亚萱. HPLC法测定血瘀证兔血清中游离阿魏酸J. 药学学报, 1995, 30(10): 762-767.
AD Wen, X Huang, YP Jiang , Yx Fan, . HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF FREE FERULIC ACID IN SERUM OF RABBITS WITH BLOOD STASISJ. Acta Pharmaceutica Sinica, 1995, 30(10): 762-767.
Citation: AD Wen, X Huang, YP Jiang , Yx Fan, . HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF FREE FERULIC ACID IN SERUM OF RABBITS WITH BLOOD STASISJ. Acta Pharmaceutica Sinica, 1995, 30(10): 762-767.

HPLC法测定血瘀证兔血清中游离阿魏酸

HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF FREE FERULIC ACID IN SERUM OF RABBITS WITH BLOOD STASIS

  • 摘要: 建立了一个用于测定正常及血瘀证兔血清中游离阿魏酸(FA)浓度的反相高效液相色谱法,此法简便、快速、灵敏而准确。以香豆素为内标,取0.2mlFA的血清,加入含有内标的乙腈沉淀蛋白,上清液挥干,残渣定容于60μl乙腈中。以DuPontZorbox C18为分析柱,流动相为乙腈-0.10mol·L-1磷酸缓冲液(pH2.5)(3:7v/v),紫外检测波长320nm,最小检出量和平均回收率分别为1ng和97.0±1.8%,血清中FA的线性范围20~800ng·ml-1(r=0.9986),日内及日间相对标准差均小于7%。本法被用于正常及血瘀证兔体内FA的药代动力学研究。通过本实验发现FA在正常及血瘀证兔体内的药代动力学参数有显著差异,与证治药效学假说基本相符。

     

    Abstract: Simple,rapid and sensitive reversed phase high performance liquid chromatographic(RP-HPLC)method for the analysis of free ferulic acid in serum of rabbits with blood stasis has been developed.Coumarin was used as internal standard.0.2 ml of serum containing ferulic acid was deproteinized with acetonitrile which contained the internal standard (coumarin).The supernatant was evaporated to dryness.The residue was dissolved in 60 μl of acetonitrile.A Du Pont Zorbox C18 column was used. A mixture of acetonitrile and 0.10 mol·L-1 phosphoric acid(3:7,v/v,pH 2.5)was used as the mobile phase ,and UV detection was performed at 320 nm.The detection limit (signal-to-noise ratio=3)and the mean recovery from serum were 1 ng and 97.0±1.8%, respectively. The method was linear within the range of 20~800 ng· ml-1(r=0.9986).The relative standard deviation were all less than 7%. The method was applied to pharmacokinetic studies in healthy rabbits and rabbits with blood stasis.Through this experiment we have found significant difference in the pharmacokinetic parameters between healthy rabbits and rabbits with blood stasis. The results obtained are coincident with syndrome and pharmacokinetics.

     

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