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Ultrasound-assisted hydrolysis of conjugated parabens in human urine and their determination by UPLC–MS/MS and UPLC–HRMS

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Abstract

Parabens are preservatives widely used in personal care products, pharmaceutical formulations as well as in food, and they are considered endocrine disruptors. For application in biomonitoring studies we developed a method for the determination of eight parabens from human urine. Sample preparation was enhanced and simplified by the combination of ultrasound-assisted enzymatic hydrolysis of conjugates (glucuronide and sulfate) followed by an extraction-free cleanup step. Quantification, using deuterated parabens as internal standards, was performed by ultrahigh-performance liquid chromatography coupled to either triple-quadrupole (UPLC–QqQ) or time-of-flight (UPLC–QqTOF) mass spectrometry. Full chromatographic separation of three butyl paraben isomers was achieved. Limits of quantification for both mass analyzers ranged from 0.1 to 0.5 μg/L for methyl, ethyl, n-/isopropyl, n-/isobutyl, and benzyl paraben in 200 μL of urine sample. The method was tested for applicability and showed high precision (intra- and interday 0.9–14.5 %) as well as high accuracy (relative recovery 95–132 %). A total of 39 urine samples were analyzed by both mass analyzers. The results agreed well, with a trend to higher deviation at low concentrations (less than 10 μg/L). Methyl, ethyl, and n-propyl paraben were detected most frequently (in more than 87 % of the samples) with median concentrations ranging from 0.8 to 16.6 μg/L. Female urine showed higher median concentrations for all parabens, which may indicate higher exposure due to lifestyle. This method permits accurate and high-throughput analysis of parabens for epidemiological studies. Further, the UPLC–QqTOF approach provides additional information on human exposure to other compounds by post-acquisition analysis.

Analytical procedure for paraben analysis from human urine samples including ultrasound-enhanced enzymatic digestion of glucuronide and sulfate conjugates, extraction-free clean-up and quantification by UHPLC-MS/MS.

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Acknowledgments

This work is part of the ASKURIS project (contract 02WRS1278A) and supported by the Federal Ministry of Education and Research (BMBF) within the funding measure RiSKWa [39]. We would like to thank Caroline Davis for supporting the optimization and method applicability experiments. We are grateful to one of the reviewers for his careful revision.

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  1. Department of Analytical Chemistry, Helmholtz Centre for Environmental Research – UFZ, Permoserstrasse 15, 04318, Leipzig, Germany

    Linda Schlittenbauer, Bettina Seiwert & Thorsten Reemtsma

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  1. Linda Schlittenbauer
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  2. Bettina Seiwert
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Correspondence to Thorsten Reemtsma.

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Schlittenbauer, L., Seiwert, B. & Reemtsma, T. Ultrasound-assisted hydrolysis of conjugated parabens in human urine and their determination by UPLC–MS/MS and UPLC–HRMS. Anal Bioanal Chem 408, 1573–1583 (2016). https://doi.org/10.1007/s00216-015-9265-2

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