A VALIDATED HPLC METHOD WITH DUAL WAVELENGTH DETECTION FOR CHLOROGENIC ACID WITH AN INTERNAL STANDARD IN PLASMA AND ITS APPLICATION IN PHARMACOKINETIC STUDIES IN RATS

摘要

A validated high performance liquid chromalographic (HPLC) method with dual wavelength detection was developed and applied to the determination of chlorogenic acid in rat plasma. Elution was performed on C_18 column at 27degC with acetronilrile-water (0.2% phosphoric acid, pH= 2.0) at different proportions according to a time scheduled programme and pumped at a flow rate of 1.0mL min~'. The eluent was delected at 325nm for chlorogenic acid and 370nmfor the internal standard, kaempferol. The inlra-day and inter-day precisions were better than 3.58% and 3.66%, respectively. The standard curve for chlorogenic acid was linear (r2- 0.9993) in the concentration range of 0.1-15 fig mL~-1. Accuracy in the measurement of samples ranged from 91.97 to 101.81%. The limit of detection and the limit of quantification for chlorogenic acid in plasma were 30 and 90 ng ml~-1 , respectively. This assay has been successfully applied in the pharmacokinelic study of chlorogenic acid through oral and intravenous administration in rats. It showed that the low bioavailabilily of chlorogenic acid and an intravenous administration route is a better dosage regimen.Tl is a simple and sensitive analytical method with good accuracy and reproducibilily. The current method also demonstrated that an internal standard with different optimal UV absorbance from that of the analyle can also be used in an analytical method.