TY - JOUR
T1 - The development and validation of a method using high-resolution mass spectrometry (HRMS) for the qualitative detection of antiretroviral agents in human blood
AU - Marzinke, Mark A.
AU - Breaud, Autumn
AU - Parsons, Teresa L.
AU - Cohen, Myron S.
AU - Piwowar-Manning, Estelle
AU - Eshleman, Susan H.
AU - Clarke, William
N1 - Funding Information:
We thank Marta Kozak, PhD of Thermo Fisher Scientific (Thermo Fisher, Waltham, MA) for antiretroviral fragment prediction analysis using Mass Frontier 7.0. We also thank Veronica Gantert and Deborah Boblitz in the collection of data for this work. The authors also thank the HPTN 052 study team and study participants for providing plasma specimens used in this study. This research was supported by the HIV Prevention Trials Network, sponsored by the National Institute of Allergy and Infectious Diseases (NIAID) , the National Institute of Mental Health (NMH) , the National Institute of Drug Abuse (NIDA) , Office of AIDS Research, of the National Institutes of Health (NIH) , and Department of Health and Human Services (DHHS) , grant UM1-AI068613 . The following reagents were obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: amprenavir, atazanavir, darunavir, efavirenz, emtricitabine, indinavir, lamivudine, lopinavir, nelfinavir, nevirapine, ritonavir, saquinavir, tenofovir, tipranavir and zidovudine.
PY - 2014/6/10
Y1 - 2014/6/10
N2 - Background: Antiretroviral drugs are used for the treatment and prevention of HIV infection. Non-adherence to antiretroviral drug regimens can compromise their clinical efficacy and lead to emergence of drug-resistant HIV. Clinical trials evaluating antiretroviral regimens for HIV treatment and prevention can also be compromised by poor adherence and non-disclosed off-study antiretroviral drug use. This report describes the development and validation of a high throughput, qualitative method for the identification of antiretroviral drugs using high-resolution mass spectrometry (HRMS) for the retrospective assessment of off-study antiretroviral drug use and the determination of potential antiretroviral therapy (ART) non-compliance. Methods: Serum standards were prepared that contained 15 antiretroviral drugs: 9 protease inhibitors (PIs), 4 nucleotide/nucleoside reverse transcriptase inhibitors (NRTIs), and 2 non-nucleoside/nucleotide reverse transcriptase inhibitors (NNRTIs). Analytical separation was achieved on a Hypersil Gold PFP (100. ×. 3. mm) column and the eluent was analyzed using the Thermo Exactive Orbitrap mass spectrometer (Exactive-MS) operated in full scan mode. Limit of identification, signal intensity precision, retention time analysis, selectivity, and carryover studies were conducted. Concordance with liquid chromatographic-tandem mass spectrometric (LC-MS/MS) methods was evaluated using remnant plasma samples from a clinical trial. Results: The limit of identification ranged from 5 to 10. ng/ml for 14 drugs (9 PIs, 1 NNRTI, 4 NRTIs) and was 150. ng/ml for 1 NNRTI. Precision studies with high and low control mixtures revealed signal intensity coefficients of variation of 3.0-27.5%. The Exactive-MS method was selective for the compounds of interest. Overall, concordance ranged from 89.1% to 100% for the screening of antiretroviral drugs in clinical plasma specimens as compared to LC-MS/MS methods. Conclusion: Using the Exactive-MS, we developed and validated a highly selective, robust method for the multiplexed detection of 15 antiretroviral drugs.
AB - Background: Antiretroviral drugs are used for the treatment and prevention of HIV infection. Non-adherence to antiretroviral drug regimens can compromise their clinical efficacy and lead to emergence of drug-resistant HIV. Clinical trials evaluating antiretroviral regimens for HIV treatment and prevention can also be compromised by poor adherence and non-disclosed off-study antiretroviral drug use. This report describes the development and validation of a high throughput, qualitative method for the identification of antiretroviral drugs using high-resolution mass spectrometry (HRMS) for the retrospective assessment of off-study antiretroviral drug use and the determination of potential antiretroviral therapy (ART) non-compliance. Methods: Serum standards were prepared that contained 15 antiretroviral drugs: 9 protease inhibitors (PIs), 4 nucleotide/nucleoside reverse transcriptase inhibitors (NRTIs), and 2 non-nucleoside/nucleotide reverse transcriptase inhibitors (NNRTIs). Analytical separation was achieved on a Hypersil Gold PFP (100. ×. 3. mm) column and the eluent was analyzed using the Thermo Exactive Orbitrap mass spectrometer (Exactive-MS) operated in full scan mode. Limit of identification, signal intensity precision, retention time analysis, selectivity, and carryover studies were conducted. Concordance with liquid chromatographic-tandem mass spectrometric (LC-MS/MS) methods was evaluated using remnant plasma samples from a clinical trial. Results: The limit of identification ranged from 5 to 10. ng/ml for 14 drugs (9 PIs, 1 NNRTI, 4 NRTIs) and was 150. ng/ml for 1 NNRTI. Precision studies with high and low control mixtures revealed signal intensity coefficients of variation of 3.0-27.5%. The Exactive-MS method was selective for the compounds of interest. Overall, concordance ranged from 89.1% to 100% for the screening of antiretroviral drugs in clinical plasma specimens as compared to LC-MS/MS methods. Conclusion: Using the Exactive-MS, we developed and validated a highly selective, robust method for the multiplexed detection of 15 antiretroviral drugs.
KW - Antiretroviral
KW - Clinical trials
KW - Exactive-MS
KW - High-resolution mass spectrometry
KW - Validation
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U2 - 10.1016/j.cca.2014.03.016
DO - 10.1016/j.cca.2014.03.016
M3 - Article
C2 - 24661980
AN - SCOPUS:84897427542
SN - 0009-8981
VL - 433
SP - 157
EP - 168
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
ER -