Development and validation of high-throughput liquid chromatography-tandem mass spectrometric method for simultaneous quantification of clopidogrel and its metabolite in human plasma.
A simple, sensitive and reliable method is described for simultaneous quantification of Clopidogrel and its metabolite in human plasma by using HTLC-MS/MS. The analytical procedure involves on-line coupling of extraction with Cyclone P (50 mm x 0.5 mm 50 microm) HTLC column by injecting 15 microL sample and chromatographic separation is performed with Cohesive Propel C18 (5 microm, 3.0 mm x 50 mm), followed by quantification with mass detector in SRM mode using ESI as an interface. The calibration curves were linear over a concentration range of 0.1-8 ng/mL of Clopidogrel and 70 ng/mL to 6 microg/mL of its metabolite using 20 mL human plasma per batch. The total run time of analysis was 7.5 min and the lower limits of quantification were 0.1 ng/mL for Clopidogrel and 70 ng/mL for its metabolite. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was applied in bioavailability and bioequivalence study.
Reddy S
Raghunadha
R
Clinical Research and Biosciences (I) Pvt. Ltd., Hyderabad, Andhra Pradesh, India. raghu_nathseelam@yahoo.co.in
Rao Divi
Koteswara
K
Chandiran
I Sarath
IS
Jayaveera
K N
KN
Naidu
Y K
YK
Reddy
M P Kalyan
MP
D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D000077144:Clopidogrel; D004355:Drug Stability; D006801:Humans; D007477:Ions; D008297:Male; D009862:Online Systems; D010975:Platelet Aggregation Inhibitors / Q000008:administration & dosage / Q000097:blood* / Q000378:metabolism* / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012371:Robotics; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013988:Ticlopidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D013997:Time Factors
D007477:Ions; D010975:Platelet Aggregation Inhibitors; D000077144:Clopidogrel; D013988:Ticlopidine