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Development and Validation of an Analytical Method for Quantitation of Bisphenol S in Rodent Plasma, Amniotic Fluid
Melanie A Rehder Silinski1, Brenda L Fletcher1, Reshan A Fernando1
1Discovery Sciences Unit, RTI International, P.O. Box 12194, Research Triangle Park, NC 27709, USA.
Abstract:
Bisphenol S (BPS) has been detected in personal care products, water, food and indoor house dust, demonstrating the potential for human exposure. Due to limited data to characterize the hazard of BPS, the National Toxicology Program (NTP) is investigating the toxicity of BPS in rodent models. Generating systemic exposure data is integral to putting toxicological findings into context. The objective of this work was to develop and validate a method to quantitate free (unconjugated parent) and total (free and all conjugated forms of) BPS in rodent plasma, amniotic fluid and fetal homogenate in support of NTP studies. The method used incubation with (total BPS) and without (free BPS) deconjugating enzyme and then protein precipitation followed by ultra-performance liquid chromatography-tandem mass spectrometry. In Sprague Dawley rat plasma, the method was linear (r ≥ 0.99) over the range 5-1,000 ng/mL, accurate (mean relative error (RE) ≤ ±10.5%) and precise (relative standard deviation (RSD) ≤ 7.7%). Mean recoveries were ≥93.1% for both free and total analyses. The limits of detection were 1.15 ng/mL (free) and 0.862 ng/mL (total) in plasma. The method was evaluated in the following study matrices: (i) male Hsd:Sprague Dawley®SD® (HSD) rat plasma, (ii) female HSD rat plasma, (iii) male B6C3F1 mouse plasma, (iv) female B6C3F1 mouse plasma, (v) HSD rat gestational day (GD) 18 dam plasma, (vi) HSD rat GD 18 amniotic fluid, (vii) HSD rat GD 18 fetal homogenate and (viii) HSD rat postnatal day 4 pup plasma (mean %RE ≤ ±8.2 and %RSD ≤ 8.7). Stability of BPS in extracted samples was demonstrated for up to 7 days at various temperatures, and freeze-thaw stability was demonstrated after three cycles over 7 days. BPS in various matrices stored at -80°C for at least 60 days was within 92.1-115% of Day 0 concentrations, demonstrating its stability in these matrices. These data demonstrate that this simple method is suitable for determination of free and total BPS in plasma, amniotic fluid and fetuses following exposure of rodents to BPS.
