LC–MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine in dried blood spots

Abstract

A simple, short, and rugged LC-MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine was developed and validated. Dried blood spots were prepared with 25 μL of spiked whole blood. A 3 mm punch was extracted with methanol containing labeled internal standards. Ten microliters was injected into the LC-MS/MS using isocratic mobile phase composed of 0.1% formic acid in water and 0.1% formic acid in acetonitrile (45: 55 v/v) at a flow rate of 0.25 mL/min. The method was validated in the range of 10-2000 ng/mL for all four analytes. The intra-assay accuracy (RE) of the method was -4.73-4.78, 1.35-2.89, -8.89 to -0.49 and - 1.40-1.81 for tenofovir, emtricitabine, elvitegravir and rilpivirine, respectively. The inter-assay accuracy was within ±15% of nominal and precision (CV) was <15%. The hematocrit effect on quantification was nonsignificant at the tested hematocrit levels (35-70%). The dried blood spot method showed good agreement with the plasma method, and hence can be used as an alternative to plasma method.

Development and validation of a multiplex UHPLC‐MS/MS assay with stable isotopic internal standards for the monitoring of the plasma concentrations of the antiretroviral drugs bictegravir, cabotegravir, doravirine, and rilpivirine in people living with HIV

“…In the only published bioassays for bictegravir, mass spectrometer operated in ESI positive mode, while in our study, bictegravir sensitivity was higher in the negative mode . In most of published methods for the quantification of rilpivirine, MS/MS transition was 367/195 in the positive mode . However, the infusion of a rilpivirine solution into the MS detector revealed a higher sensitivity in the negative mode.…”

Detection and quantification of Covid-19 antiviral drugs in biological fluids and tissues

“…blood, plasma, breast milk) spots [ [34] , [35] , [36] , [37] , [38] , [39] , [40] , [41] ]. However, a lack of standard validation guidelines for drug quantification in dried samples spots is ascertained [ 34 ]. Therefore, accepted bio-analytical methodology validation practises are typically followed in those cases.…”

“…A separate discussion is needed for the preparation of dried sample spots [ [34] , [35] , [36] , [37] , [38] , [39] , [40] , [41] ]. This low-cost technology has recently received a great deal of attention because of their distinct advantages of sample collection, less invasive sampling, simpler transfer, storage and shipping [ 104 , 105 ].…”

“…Before the analysis, they are placed in clean tubes, where the extraction solution is added. The extraction can be assisted by sonication [ 34 , 38 ] and, after centrifugation, the supernatant is collected for chromatographic analysis.…”