TY - JOUR
T1 - LC–MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine in dried blood spots
AU - Prathipati, Pavan Kumar
AU - Mandal, Subhra
AU - Destache, Christopher J.
N1 - Funding Information:
This work was supported by NIH grant RO1 AI117740‐01 to C.J.D. The Animal Research Facility is supported by grant number G20RR024001 from the National Center for Research Resources. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Research Resources or the National Institutes of Health.
Publisher Copyright:
Copyright © 2018 John Wiley & Sons, Ltd.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 2018/9
Y1 - 2018/9
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85047484947&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85047484947&partnerID=8YFLogxK
U2 - 10.1002/bmc.4270
DO - 10.1002/bmc.4270
M3 - Article
AN - SCOPUS:85047484947
VL - 32
JO - Biomedical Chromatography
JF - Biomedical Chromatography
SN - 0269-3879
IS - 9
M1 - e4270
ER -