Optimized extraction and analysis methods using liquid chromatography-tandem mass spectrometry for zearalenone and metabolites in human placental tissue
Abigail Lazofsky,
Anita Brinker,
Ruby Gupta,
Emily Barrett,
Lauren M. Aleksunes,
Zorimar Rivera-Núñez,
Brian Buckley
Affiliations
Abigail Lazofsky
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ, 08854, USA
Anita Brinker
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ, 08854, USA
Ruby Gupta
Department of Environmental and Occupational Health and Justice, Rutgers School of Public Health, Rutgers University, 683 Hoes Lane West, Piscataway, NJ, 08854, USA
Emily Barrett
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ, 08854, USA; Department of Biostatistics and Epidemiology, Rutgers School of Public Health, Rutgers University, 683 Hoes Lane West, Piscataway, NJ, 08854, USA
Lauren M. Aleksunes
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ, 08854, USA; Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, 160 Frelinghuysen Road, Rutgers University, Piscataway, NJ, 08854, USA; Rutgers Center for Lipid Research, New Jersey Institute for Food, Nutrition, and Health, Rutgers University, 61 Dudley Road, New Brunswick, NJ, 08901, USA
Zorimar Rivera-Núñez
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ, 08854, USA; Department of Biostatistics and Epidemiology, Rutgers School of Public Health, Rutgers University, 683 Hoes Lane West, Piscataway, NJ, 08854, USA
Brian Buckley
Environmental and Occupational Health Sciences Institute, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ, 08854, USA; Corresponding author.
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.
