Universitas Indonesia Conferences, 1st International Conference on Advance Pharmacy and Pharmaceutical Sciences

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Analytical Method Validation Clopidogrel in Human Plasma by Ultra Performance Liquid Chromatography - Mass Spectrometer
Yahdiana Harahap, Innet Maysyarah, Herman Suryadi

Last modified: 2016-11-08


Clopidogrel is an antiplatelet drug with a very small plasma concentration because of its extensive metabolism into active and inactive metabolites after oral administration. The parent clopidogrel has plasma concentration 2000 times lower than the inactive metabolite, clopidogrel carboxylic. This study was aimed to obtain sensitive, selective and valid method to analyze  clopidogrel in plasma, using Ultra Performance Liquid Chromatography tandem Mass Spectrometry (UPLC-MS/MS). Chromatography system used are Waters Acquity UPLC Class BEH C18 1.7 µm ( 2.1 x 100 mm) column and 0.1% formic acid in water – 0.1% formic acid in acetonitrile (30-70) as mobile phase, under isocratic elution with flow rate  0.2 mL/min. Mass detection was performed with Waters Xevo TQD equipped with positive electrospray ionization (ESI) on multiple reaction monitoring (MRM) mode. Clopidogrel was detected at m/z 322.086 > 212.097, compared to internal standard irbesartan  at m/z 429.233 > 207.131. Sample was prepared by protein precipitation method with acetonitrile, mixed with vortex for 10 minutes, and then centrifuged on 13000 rpm for 20 minutes. This method is showing linear result at the concentration range 0.2-10 ng/mL with r  > 0.9997. This method meets the 2011 EMEA Bioanalytical Guideline validation requirement.