Objective:To establish a method of HPLC-Fluorescence to determine beagle dog plasma concentration of Adefovir

parent drug of analogue of Adefovir dipivoxil-M1

M2.Methods:The analyte was separated on a Benetnach C18 column(150×4.6 mm

5 μm).The mobile phase were:A

2% acetonitrile in 25 mmol potassium phosphate buffer with 5 mmol tetrabutylammonium hydroxide

pH 6.0;B

65% acetonitrile in 25 mmol potassium phosphate buffer with 5 mmol tetrabutylammonium hydroxide

pH 6.0.The flow rate was 1.0 ml/min and the column temperature was maintained at 35℃.The injection volume was 10 μl.Results:The calibration curve was linear over plasma concentration range of 5.0～750.0 ng/ml

r=0.999 8.The lower limit of quantification in plasma of beagle dogs was 2.0 ng/ml.Relative recovery of Adefovir in plasma of beagle dogs was 98.36%～99.58%

and absolute recovery was 79.76%～82.14%.The interday precision(RSD%) in plasma was 2.12%～3.77%

and the intraday precision(RSD%) was 3.07%～5.96%.Conclusion:The established method has a good selectivity and reproducibility

and is suitable for the determination of Adefovir in plasma of beagle dogs.