TY - JOUR
T1 - Development and validation of a multiplexed assay for the measurement of long-acting hormonal contraceptives in plasma via liquid chromatography-tandem mass spectrometry
AU - Knezevic, Claire E.
AU - Parsons, Teresa L.
AU - Gollings, Ryann
AU - Pandey, Aashish
AU - Marzinke, Mark A.
N1 - Funding Information:
This work was supported by the HIV Prevention Trials Network Laboratory Center , sponsored by the National Institute of Allergy and Infectious Diseases ( NIAID ), the National Institute of Mental Health ( NIMH ), and the National Institute of Drug Abuse ( NIDA ), Office of AIDS Research, of the National Institutes of Health ( NIH ), Department of Health and Human Services ( DHHS ), grant UM1-AI068613 . The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. This work was partially supported by the Johns Hopkins University Center for AIDS Research ( CFAR ; P30AI094189 ).
Publisher Copyright:
© 2023 Elsevier B.V.
PY - 2023/5/10
Y1 - 2023/5/10
N2 - Background: Exogenous progestins are an effective tool for hormonal contraception and family planning. Progestins may be delivered as oral pills, intramuscular or subcutaneous injections, vaginal rings, or intrauterine devices. Drug concentrations may vary based on the route and duration of delivery. Measurement of synthetic steroids in blood plasma can aid in determination of product adherence, evaluation of drug-drug interactions, and investigation of unintended pregnancies. Methods: Drug-free K2EDTA plasma was spiked with the synthetic steroids etonogestrel (ETO), levonorgestrel (LNG), medroxyprogesterone acetate (MPA), and norethisterone (NET). Plasma was combined with isotopically labeled internal standards, and drugs were extracted via liquid-liquid extraction. Samples were then subjected to liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. The method was validated in accordance with regulatory recommendations. The assay was evaluated in a cohort of remnant plasma samples in individuals using one of the aforementioned progestins. Results: The analytical measuring range for ETO, MPA, and NET was 20–10,000 pg/mL; the primary linearity for LNG was 20–20,000 pg/mL. The method showed acceptable precision and accuracy for all progestins. Stability was established for 72 h with room temperature storage and through 3 freeze-thaw cycles. All analytes were stable in whole blood incubated at room temperature for 25 h, and at 40°C and 100% humidity for 2 h. Ion suppression was observed for all analytes spiked in plasma; average ion suppression was 31.6%, 66.6%, 32.1% and 41.2% for ETO, LNG, MPA, and NET, respectively. However, internal standards showed comparable ion suppression, and relative matrix effects were minimal. ETO, LNG, MPA, and NET could also be quantified accurately in K3EDTA plasma and serum. Progestins were successfully measured in remnant samples from individuals using hormonal contraceptives. Conclusions: A multiplexed LC-MS/MS assay for the quantification of ETO, LNG, MPA, and NET has been developed and validated. The assay met acceptable performance characteristics and may be used in downstream studies to evaluate progestin pharmacology.
AB - Background: Exogenous progestins are an effective tool for hormonal contraception and family planning. Progestins may be delivered as oral pills, intramuscular or subcutaneous injections, vaginal rings, or intrauterine devices. Drug concentrations may vary based on the route and duration of delivery. Measurement of synthetic steroids in blood plasma can aid in determination of product adherence, evaluation of drug-drug interactions, and investigation of unintended pregnancies. Methods: Drug-free K2EDTA plasma was spiked with the synthetic steroids etonogestrel (ETO), levonorgestrel (LNG), medroxyprogesterone acetate (MPA), and norethisterone (NET). Plasma was combined with isotopically labeled internal standards, and drugs were extracted via liquid-liquid extraction. Samples were then subjected to liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. The method was validated in accordance with regulatory recommendations. The assay was evaluated in a cohort of remnant plasma samples in individuals using one of the aforementioned progestins. Results: The analytical measuring range for ETO, MPA, and NET was 20–10,000 pg/mL; the primary linearity for LNG was 20–20,000 pg/mL. The method showed acceptable precision and accuracy for all progestins. Stability was established for 72 h with room temperature storage and through 3 freeze-thaw cycles. All analytes were stable in whole blood incubated at room temperature for 25 h, and at 40°C and 100% humidity for 2 h. Ion suppression was observed for all analytes spiked in plasma; average ion suppression was 31.6%, 66.6%, 32.1% and 41.2% for ETO, LNG, MPA, and NET, respectively. However, internal standards showed comparable ion suppression, and relative matrix effects were minimal. ETO, LNG, MPA, and NET could also be quantified accurately in K3EDTA plasma and serum. Progestins were successfully measured in remnant samples from individuals using hormonal contraceptives. Conclusions: A multiplexed LC-MS/MS assay for the quantification of ETO, LNG, MPA, and NET has been developed and validated. The assay met acceptable performance characteristics and may be used in downstream studies to evaluate progestin pharmacology.
KW - Etonogestrel
KW - Hormonal contraceptives
KW - LC-MS
KW - Levonorgestrel
KW - Mass spectrometry
KW - Medroxyprogesterone acetate
KW - Norethisterone
KW - Progestins
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U2 - 10.1016/j.jpba.2023.115321
DO - 10.1016/j.jpba.2023.115321
M3 - Article
C2 - 36924631
AN - SCOPUS:85150037797
SN - 0731-7085
VL - 228
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
M1 - 115321
ER -