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 C18(ODS2), column (150 mm x 4.6 mm, 5 microns), 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 micrograms/L, with a linearity of 33.7-2156.8 micrograms/L, r = 0.9993. The mean recovery of serum FA was (93.59 +/- 2.36) micrograms/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.