METHOD VALIDATION FOR QUANTIFICATION OF LOPINAVIR IN HUMAN PLASMA BY USING LC-MS/MS
European Journal of Molecular & Clinical Medicine,
2023, Volume 10, Issue 1, Pages 1115-1133
AbstractA satisfactory LC-MS/MS technique was designed and validated in accordance with FDA standards for the quantification of lopinavir in plasma samples employing verapamil as IS. The designing of LC-MS/MS technique utilized a variety of commonly accessible UPLC columns and different mobile phases. Optimal separation was achieved when Waters X bridge C18 column (100 mm x 2.1 mm ID, 3.5 μm) was utilized along with mobile phase comprising of 2 mM AFB with 0.1% formic acid: ACN in the proportion of 20:80, v/v at 0.3 mL/min. ESI-MS at +ve ion mode was employed for detecting the sample. For quantitation, the MRM ratios for lopinavir (m/z 629.85 to 183.26) and verapamil (m/z 455.49 to 165.04) were employed. Verapamil and lopinavir each had retention times of 1.01 and 1.85 minutes, correspondingly. The pearson correlation (r=0.997) was used to confirm linearity for lopinavir around the concentration ranging between 2 ng/mL and 1000 ng/mL, and the total recoveries were 98.28% and 96.47% for lopinavir and verapamil, correspondingly. The presented approach also has a shorter analytic run time than earlier published methods, which is a benefit. As a result, the technique is appropriate for Lopinavir estimation.
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