TY - JOUR
T1 - Optimized extraction and analysis methods using liquid chromatography-tandem mass spectrometry for zearalenone and metabolites in human placental tissue
AU - Lazofsky, Abigail
AU - Brinker, Anita
AU - Gupta, Ruby
AU - Barrett, Emily
AU - Aleksunes, Lauren M.
AU - Rivera-Núñez, Zorimar
AU - Buckley, Brian
N1 - Publisher Copyright: © 2023 The Authors
PY - 2023/6
Y1 - 2023/6
N2 - Zearalenone and its metabolites, a group of endocrine disrupting mycotoxins, have been linked to adverse reproductive health effects. They cross the placental barrier, potentially reaching the fetus. In this study, we adapted and optimized our protocol previously used for urine, to measure these mycotoxins in human placentas. We combined a supported liquid extraction step using Chem Elut cartridges with solid phase extraction on Discovery® DSC-NH2 tubes. The optimized extraction efficiencies were between 68 and 80% for all metabolites. Analysis was performed by UHPLC-HRMS using a Betasil™ Phenyl-Hexyl column eluted with a gradient of acetonitrile-methanol-water. The chromatography method separated all analytes in under 15 min. Validation experiments confirmed the method's sensitivity, with LODs ranging from 0.0055 to 0.011 pg/mg tissue. The method was linear over a range of 0.0025–1.5 pg/mg tissue with R2 values ≥ 0.994. Precision and accuracy calculations ranged from 4.7–7.9% and 0.6–6.7% respectively. The method was then successfully applied to a subset of placenta samples (n = 25) collected from an ongoing prospective birth cohort. Interestingly, 92% of the samples contained at least one measurable zearalenone metabolite, providing initial indication of potentially widespread exposure during pregnancy.
AB - Zearalenone and its metabolites, a group of endocrine disrupting mycotoxins, have been linked to adverse reproductive health effects. They cross the placental barrier, potentially reaching the fetus. In this study, we adapted and optimized our protocol previously used for urine, to measure these mycotoxins in human placentas. We combined a supported liquid extraction step using Chem Elut cartridges with solid phase extraction on Discovery® DSC-NH2 tubes. The optimized extraction efficiencies were between 68 and 80% for all metabolites. Analysis was performed by UHPLC-HRMS using a Betasil™ Phenyl-Hexyl column eluted with a gradient of acetonitrile-methanol-water. The chromatography method separated all analytes in under 15 min. Validation experiments confirmed the method's sensitivity, with LODs ranging from 0.0055 to 0.011 pg/mg tissue. The method was linear over a range of 0.0025–1.5 pg/mg tissue with R2 values ≥ 0.994. Precision and accuracy calculations ranged from 4.7–7.9% and 0.6–6.7% respectively. The method was then successfully applied to a subset of placenta samples (n = 25) collected from an ongoing prospective birth cohort. Interestingly, 92% of the samples contained at least one measurable zearalenone metabolite, providing initial indication of potentially widespread exposure during pregnancy.
KW - Biomarker
KW - Mycoestrogens
KW - Mycotoxins
KW - Placenta
KW - Solid-phase extraction
KW - Zeranols
UR - http://www.scopus.com/inward/record.url?scp=85161297942&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85161297942&partnerID=8YFLogxK
U2 - 10.1016/j.heliyon.2023.e16940
DO - 10.1016/j.heliyon.2023.e16940
M3 - Article
SN - 2405-8440
VL - 9
JO - Heliyon
JF - Heliyon
IS - 6
M1 - e16940
ER -