The Development and Validation of an UHPLC–MS/MS Method for The Rapid Quantification of The Antiretroviral Agent Dapivirine in Human Plasma

Abstract

Background: Dapivirine is a non-nucleoside reverse transcriptase inhibitor designed to prevent HIV-1 viral replication and subsequent propagation. A sensitive method is required to quantify plasma concentrations to assess drug efficacy. Results: Dapivirine-spiked plasma was combined with acetonitrile containing deuterated IS and was processed for analysis. The method has an analytical measuring range from 20 to 10,000 pg/ml. For the LLOQ, low, mid and high QCs, intra- and inter-assay precision (%CV) ranged from 5.58 to 13.89% and 5.23 to 13.36%, respectively, and intra- and inter-day accuracy (% deviation) ranged from -5.61 to 0.75% and -4.30 to 6.24%, respectively. Conclusion: A robust and sensitive LC–MS/MS assay for the high-throughput quantification of the antiretroviral drug dapivirine in human plasma was developed and validated following bioanalytical validation guidelines. The assay meets criteria for the analysis of samples from large research trials.

Financial & competing interests disclosure

This work was supported by the Microbicide Trials Network sponsored by the NIH/Division of AIDS grant UM1 AI068633. We are also grateful for essential analytical equipment provided by a grant from the James B. Pendleton Charitable Trust. CW Hendrix received partial support through a contract between Gilead Sciences, Inc. and The Johns Hopkins University. Gilead Sciences, Inc., also markets the tenofovir disoproxil fumarate-containing products Viread^®, Truvada^® and Stribild^®. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Acknowledgements

The following reagents were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, National Institute of Allergies and Infectious Diseases, NIH: dapivirine.

Additional information

Funding

This work was supported by the Microbicide Trials Network sponsored by the NIH/Division of AIDS grant UM1 AI068633. We are also grateful for essential analytical equipment provided by a grant from the James B. Pendleton Charitable Trust. CW Hendrix received partial support through a contract between Gilead Sciences, Inc. and The Johns Hopkins University. Gilead Sciences, Inc., also markets the tenofovir disoproxil fumarate-containing products Viread^®, Truvada^® and Stribild^®. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.