Abstract
Ortho-phenylphenol (OPP) has been widely used as a fungicide and preservative. Although low-dose studies have demonstrated its low toxicity in animals and humans, high-dose exposure to this contaminant has toxic effects that range from skin irritation to bladder cancer. Thus far, monitoring of OPP exposure in the general population has been performed by measuring OPP after urine hydrolysis with the β-glucuronidase/arylsulfatase enzyme and sometimes by the use of a mineral acid. We developed a sensitive, accurate, and robust method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to specifically measure two-phase II OPP metabolites excreted in human urine, OPP sulfate (OPP-S), and OPP glucuronide (OPP-G). Comparative analysis of urine samples from 50 volunteers living in the Quebec City area using a direct method and phosphoric acid hydrolysis method previously developed in our laboratory showed no statistically significant difference (p value for paired t test = 0.701) in OPP concentrations. Moreover, a significant difference showed that underestimation (p value for paired t test = 0.025) occurs when β-glucuronidase/arylsulfatase enzyme deconjugation is used. The LOD achieved by the direct method permits the detection of OPP-S and OPP-G metabolites in urine at the submicrogram per liter level.
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Acknowledgments
We thank all the donors for kindly providing urine samples for this study. We are also very grateful to Nathalie Morissette, Nancy Plamondon, and Claudine Roussy for their technical support to the method development and validation. We acknowledge Sébastien Gagné for his technical help to prepare the manuscript and Dr. Pierre Ayotte for reviewing.
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Financial support was provided by an internal budget dedicated to development and research.
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This study involves the use of human urine samples collected from multiple volunteer donors. We obtained the verbal consent of all participants to use the data obtained from those samples for research purposes. For donors under the age of 16, informed consent was obtained from the parents of these participants. Urine samples were exclusively used for method development and validation. The samples were coded to maintain the anonymity of the participants and disconnect the identity of each donor from the data generated from their urine sample. The study was performed in accordance with the ethical standards.
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Bérubé, R., Bélanger, P., Bienvenu, JF. et al. New approach for the determination of ortho-phenylphenol exposure by measurement of sulfate and glucuronide conjugates in urine using liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 410, 7275–7284 (2018). https://doi.org/10.1007/s00216-018-1339-5
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DOI: https://doi.org/10.1007/s00216-018-1339-5