High-performance liquid chromatographic determination of inactive carboxylic acid metabolite of clopidogrel in human serum: Application to a bioequivalence study.
A sensitive and rapid method is described for determination of clopidogrel carboxylic acid (CCA), the inactive metabolite of anti platelet agent, clopidogrel, in human serum. The analytical procedure involves liquid-liquid extraction of the analyte and an internal standard (phenytoin) with ethyl acetate. A mobile phase consisting of 0.05 M phosphate buffer containing triethylamine (0.5 mL/L; pH 5.7) and acetonitrile (56:44 v/v) was used and chromatographic separation was achieved using C18 analytical column at detector wavelength of 220 nm. The calibration curves were linear over a concentration range of 0.05-10 microg/mL of CCA in human serum. The total run time of analysis was 5.5 min and the lower limits of detection (LOD) and quantification (LOQ) were 0.02 and 0.05 microg/mL, respectively. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was applied in a randomized cross-over bioequivalence study of two different clopidogrel preparations in 24 healthy volunteers.
Bahrami
Gholamreza
G
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran. gbahrami@kums.ac.ir <gbahrami@kums.ac.ir>
Mohammadi
Bahareh
B
Sisakhtnezhad
Sajjad
S
D000097:Acetonitriles; D000328:Adult; D002264:Carboxylic Acids / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000077144:Clopidogrel; D004355:Drug Stability; D005021:Ethylamines; D006801:Humans; D008297:Male; D010975:Platelet Aggregation Inhibitors / Q000097:blood / Q000493:pharmacokinetics; D012680:Sensitivity and Specificity; D013810:Therapeutic Equivalency; D013988:Ticlopidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics
D000097:Acetonitriles; D002264:Carboxylic Acids; D005021:Ethylamines; D010975:Platelet Aggregation Inhibitors; D000077144:Clopidogrel; D013988:Ticlopidine; C016162:triethylamine; C032159:acetonitrile