Method development and validation of repaglinide in human plasma by HPLC and its application in pharmacokinetic studies.
In this study, the development and validation of a high-performance liquid chromatography (HPLC) assay for determination of repaglinide concentration in human plasma for pharmacokinetic studies is described. Plasma samples containing repaglinide and an internal standard, indomethacin were extracted with ethylacetate at pH 7.4. The recovery of repaglinide was 92%+/-55.31. Chromatographic separations were performed on Purospher STAR C-18 analytical column (4.8 mm x 150 mm; 5 microm particle size). The mobile phase composed of acetonitrile-ammonium formate (pH 2.7; 0.01 M) (60:40, v/v). The flow rate was 1 ml/min. The retention time for repaglinide and indomethacin were approximately 6.2 and 5.3 min, respectively. Calibration curves of repaglinide were linear in the concentration range of 20-200 ng/ml in plasma. The limits of detection and quantification were 10 ng/ml and 20 ng/ml, respectively. The inter-day precision was from 5.21 to 11.84% and the intra-day precision ranged from 3.90 to 6.67%. The inter-day accuracy ranged 89.95 to 105.75% and intra-day accuracy ranged from 92.37 to 104.66%. This method was applied to determine repaglinide concentration in human plasma samples for a pharmacokinetic study.
Ruzilawati
Abu Bakar
AB
Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kelantan, Malaysia. tieimran@yahoo.com
Wahab
Mohd Suhaimi Abd
MS
Imran
Ahmad
A
Ismail
Zabidah
Z
Gan
Siew Hua
SH
D002219:Carbamates / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004355:Drug Stability; D005615:Freezing; D006801:Humans; D007004:Hypoglycemic Agents / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D015394:Molecular Structure; D010880:Piperidines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002219:Carbamates; D007004:Hypoglycemic Agents; D010880:Piperidines; C072379:repaglinide