Abstract
Objective: This study developed a sensitive, selective, and valid method for analyzing clopidogrel in human plasma using liquid chromatographytandemmass spectrometry (LC-MS/MS).Methods: The chromatography separation was performed on the waters ACQUITY UPLC Class BEH C18 1.7 μm (2.1 × 100 mm) column, consisting of0.1% formic acid in water and 0.1% formic acid in acetonitrile (30-70) as the mobile phase, and an isocratic elution with a flow rate of 0.2 mL/minutes.The mass detection was performed using a Waters Xevo TQD equipped with positive electrospray ionization in the multiple reaction monitoringmodes. The clopidogrel was detected at m/z 322.086>212.097 and irbesartan as an internal standard at m/z 429.233>207.131. The sample wasprepared with protein precipitation method using acetonitrile, vortex mixed for 10 minutes, and centrifuged at 13,000 rpm for 20 minutes.Results: This method showed a linear result at the concentration range of 0.2-10 ng/ml with r≥0.9997.Conclusions: The developed method provides sensitivity, linearity, precision, and accuracy and is suitable for analysis of clopidogrel in human plasmausing LC-MS/MS.
Highlights
Clopidogrel is used for the secondary prevention of cerebral or myocardial infarction in patients who cannot tolerate acetylsalicylic acid or those who undergo a cerebral or myocardial infarction while taking acetylsalicylic acid [1]
Clopidogrel is a prodrug, with a slow onset, that has been found to be as effective as acetylsalicylic acid for secondary prevention in stroke
In 2012, another study conducted a validation of the clopidogrel analytical method through protein precipitation, which was simpler and did not use methanol in the analysis [8]
Summary
The chromatography separation was performed on the waters ACQUITY UPLC Class BEH C18 1.7 μm (2.1 × 100 mm) column, consisting of 0.1% formic acid in water and 0.1% formic acid in acetonitrile (30-70) as the mobile phase, and an isocratic elution with a flow rate of 0.2 mL/minutes. The mass detection was performed using a Waters Xevo TQD equipped with positive electrospray ionization in the multiple reaction monitoring modes. The clopidogrel was detected at m/z 322.086>212.097 and irbesartan as an internal standard at m/z 429.233>207.131. The sample was prepared with protein precipitation method using acetonitrile, vortex mixed for 10 minutes, and centrifuged at 13,000 rpm for 20 minutes
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