Method development, validation, and application of liquid chromatography-electrospray ionization-mass spectrometry/mass spectrometry method for the quantification of amprenavir in plasma samples

Abstract

A specific liquid chromatography-mass spectrometry/mass spectrometry spectrometric procedure for the quantitation of amprenavir drug in biological matrices was developed and validated. Chromatographic isolation was accomplished through a Zorbax C18 analytical stationary phase having the dimensions of 50 mm × 4.6 mm and particle size of 5.0 μm. Isocratic separation was processed with acetonitrile 0.1%v/v HCOOH in water and methyl alcohol in the proportion of 60:10:30 as a moveable system with a flow rate of 0.60 mL/min. Liquid-liquid extraction was carried out for drug and internal standard isolation with an ethyl acetate solvent. Parent and product ionic components were examined at m/z 506.2 → 89.1 for amprenavir and 367.1 → 350.1 for rilpivirine internal standard on the MRM (multiple reaction monitoring) mode. The linearity plot of analyte was rectilinear in the concentration over 0.15-1500 ng/mL with the correlation coefficient value of r 2 being >0.990. %relative standard deviation findings were <4.21% for intraday and interday accuracy and precision. The technique has good recoveries, and %recovery findings of LQC (low quality control), MQC(median quality control), and HQC (high quality control) solutions were 92.9%, 95.1%, and 96.4%, respectively. Amprenavir has more stability for longer time when subjected to different stability environments and the procedure was efficiently relevant to the regular investigation of amprenavir analyte in the biological matrix.

An LC-MS Method Development and Validation for the Estimation of Ritonavir in Plasma Samples

“…We obtained plasma (biological matrix) from at least 8 individual batches, including one lipemic and one hemolytic plasma. 15 In order to assess interference, the response in the blank matrix will be compared to the average result of the extracted LLoQ for the drug at the respective retention time (RT) values (Figures 2 to 4). Similarly, the response in the blank matrix will be compared to the average result of the extracted 2).…”