Determination of AKF-PD in whole blood of rat by HPLC-UV

An HPLC-UV method was developed and validated for the determination of AKF-PD in whole blood of rat. Phenacetin was chosen as the internal standard, and the separation was achieved on a C18 column with methanol and 0.02 M phosphate buffer (pH 3.2) as mobile phase. The obtained calibration graphs were linear (r = 0.9999, n = 9) in the range of 0.203–52.0 μg ml−1. The low limit of quantitation was 0.203 μg ml−1. This method can be used to study the pharmacokinetics of AKF-PD in rat.