高效液相色谱直接测定沸水浴处理人血清中冠心Ⅱ号来源的阿魏酸

Direct Determination of Serum Content of Ferulic Acid Treated with Boiling Water by HPLC in Vonlunteers after Oral Administration of Coronary Heart No. Ⅱ Decoction

目的：探讨建立健康人口服冠心Ⅱ号后吸收入血清成分的临床药动学（ＰＫ）研究的新方法。方法：用高效液相色谱（ＨＰＬＣ）直接测定５名健康人服冠心Ⅱ号后经沸水浴处理过的血清样品中的阿魏酸（ＦＡ）。分别用二维、三维ＨＰＬＣ和紫外分光光度法定性ＦＡ后，ＦＡ用内标（香豆精）法定量。结果：甲醇∶乙酸∶水（３８∶０３∶６１７）为流动相，Ｃ１８（ＯＤＳ２）柱（１５０ｍｍ×４６ｍｍ，５μｍ）为固定相时，ＦＡ的最低检测限６ｎｇ（Ｎ／Ｓ＝３）；沸水浴１０ｍｉｎ的最低检测血清浓度是２０２μｇ／Ｌ，线性范围３３７～２１５６８μｇ／Ｌ，ｒ＝０９９９３；方法平均回收率（９３５９±２３６）％，日间及日内精密度ＲＳＤ值均＜８４４％。结论：与乙睛法比较，本法灵敏、快速、简便、精确、专一、价廉、无毒和重复性好。

Objective: To develop a new method for clinical pharmacokinetic detection of chemical component of Coronary Heart NO. Ⅱ in serum after oral administration in healthy person. Methods: Content of ferulic acid (FA) in serum ,which had been treated with boiling water bath, was determined directly by high performance liquid chromotography (HPLC) and spectrophotometer. After qualitative detection performed by using Bi and tri dimensional HPLC and ultraviolet spectrophotometer, the quantitative detection was determined with the internal standard (coumarin). Results: When eluted by mobile phase of a mixture of methanol, acetic acid and water (38∶0.3∶61.7 in ratio), and stationary phase of C 18 (ODS 2), column (150mm×4.6mm, 5μm), the minimal detectable limit of free methanol FA was 6 ng (N/S=3), and the minimal detectable serum FA concentration, after 10 min of boiling water bath, was 20.2 μg/L, with a linearity of 33.7～2156.8 μg/L, r=0.9993. The mean recovery of serum FA was (93.59±2.36) μg/L. The RSD within day and day to day were all less than 8.44%. Conclusion: In comparing with sample Preparation of acetonitrile, this method is simple, rapid, sensitive, cheap, accurate, specific, reproducible and no toxic.