Development of an Analytical Method for Quantitation of Deoxynivalenol by UPLC-MS-MS: A Preliminary Assessment of Gestational and Lactational Transfer in Rats.

Deoxynivalenol (DON) is the most widely distributed trichothecene mycotoxin in grain-based foods and animal feed. Exposure to DON is widespread as it has been detected in food sources from around the world. The objective of this work was to develop a method to quantitate DON in biological matrices and apply it in a preliminary assessment of gestational and lactational transfer of DON following exposure of pregnant rats. The method used protein precipitation followed by ultra-performance liquid chromatography-tandem mass spectrometry. The method was evaluated in male Sprague Dawley rat plasma over the concentration range ∼2-1,000 ng/mL. The method was linear (r ≥ 0.99), accurate (mean relative error ≤ ±4.9%) and precise (relative standard deviation ≤ 5.5%). The mean absolute recovery was 85.9%. The limit of detection was 0.35 ng/mL. The method was also evaluated in gestational day (GD) 18 Hsd:Sprague Dawley®SD® dam plasma and fetal homogenate (mean % relative error ≤ ±16.9; % relative standard deviation ≤ 9.5). Concentrations of DON in dam plasma stored at -80°C for at least 29 days and in fetal homogenate for at least 43 days were within 97.9 to 120% of Day 0 concentrations, demonstrating that DON is stable in these matrices. The method was used to quantitate DON in rat maternal plasma, amniotic fluid, GD 18 fetuses and postnatal day (PND) 4 pups following exposure of dams to 0 (control) and 1 mg/kg DON beginning on GD 6 and continuing through gestation and lactation for a preliminary assessment of maternal transfer. In animals exposed to 1 mg/kg/day, similar concentration of DON was found in GD 18 dam plasma and fetuses, demonstrating significant gestational transfer. The concentration of DON in PND 4 dam plasma was similar to that in GD 18 dam plasma. However, DON was not detected in PND 4 pup plasma above the limit of detection of the assay, demonstrating absence of transfer of DON to pups via lactation.