Optimization and validation of a fast RP-HPLC method for the determination of dobutamine in rat plasma:Pharmacokinetic studies in healthy rat subjects  

作　　者：Ramesh Thippani Nageswara Rao Pothuraju Nageswara Rao Ramisetti Saida Shaik 

机构地区：[1]Department of Chemistry,National Institute of Technology [2]Analytical Chemistry Division,IICT

出　　处：《Journal of Pharmaceutical Analysis》2013年第6期434-439,共6页药物分析学报（英文版）

基　　金：Mr.Thippani Ramesh thanks MHRD,Government of India for providing financial assistance

摘　　要：A novel isocratic reverse phase high performance liquid chromatography （RP-HPLC） with photo diode array （PDA） detection method for the determination of dobutamine （DBT） in rat plasma was developed and validated after optimization of various chromatographic conditions and other experimental parameters. Homoveratrylamiue was used as an internal standard. Methanol was used as the extracting solvent for the preparation of plasma samples. Samples were separated on a Symmetry C18 （250ram x4.6mm i.d., 5 pro） analytical column. Acetonitrile and 15raM potassium dihydrogen phosphate （pH 5.0 with 0.3% TEA） （20：80, v/v） was used. The column oven temperature was optimized at 35 ~C and the flow rate was 0.8 mL/min. The detection wavelength was fixed at 230 nm for entire analysis. The calibration curve was found to be linear over the concentration range of 50-2000 ng/mL （ra=0.9992）. The limit of quantification （LOQ） of the method was 50 ng/mL. The % RSD values of accuracy and precision values for intra and inter days were 〈 15% at quality control （QC） concentrations. Recovery, stability and robustness were studied within the acceptable range according to ICH guidelines. The method was efficiently applied to a pharmacokinetic study in healthy Wistar rats.A novel isocratic reverse phase high performance liquid chromatography （RP-HPLC） with photo diode array （PDA） detection method for the determination of dobutamine （DBT） in rat plasma was developed and validated after optimization of various chromatographic conditions and other experimental parameters. Homoveratrylamiue was used as an internal standard. Methanol was used as the extracting solvent for the preparation of plasma samples. Samples were separated on a Symmetry C18 （250ram x4.6mm i.d., 5 pro） analytical column. Acetonitrile and 15raM potassium dihydrogen phosphate （pH 5.0 with 0.3% TEA） （20：80, v/v） was used. The column oven temperature was optimized at 35 ~C and the flow rate was 0.8 mL/min. The detection wavelength was fixed at 230 nm for entire analysis. The calibration curve was found to be linear over the concentration range of 50-2000 ng/mL （ra=0.9992）. The limit of quantification （LOQ） of the method was 50 ng/mL. The % RSD values of accuracy and precision values for intra and inter days were 〈 15% at quality control （QC） concentrations. Recovery, stability and robustness were studied within the acceptable range according to ICH guidelines. The method was efficiently applied to a pharmacokinetic study in healthy Wistar rats.

关 键 词：DOBUTAMINE RP-HPLC VALIDATION Rat plasma PHARMACOKINETICS 

分 类 号：R965[医药卫生—药理学]