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    A study of inter-individual variability in the Phase II metabolism of xenobiotics in human skin

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    Publication date
    2018-08
    Author
    Spriggs, S.
    Cubberley, R.
    Loadman, Paul M.
    Sheffield, D.
    Wierzbicki, Antonia
    Keyword
    Phase II metabolism; Ex vivo skin; S9 incubations; Cofactors
    Rights
    © 2018 Elsevier B.V. Reproduced in accordance with the publisher's selfarchiving policy. This manuscript version is made available under the CC-BY-NC-ND 4.0 license.
    Peer-Reviewed
    Yes
    
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    Abstract
    Understanding skin metabolism is key to improve in vitro to in vivo extrapolations used to inform risk assessments of topically applied products. However, published literature is scarce and usually covers a limited and non-representative number of donors. We developed a protocol to handle and store ex vivo skin samples post-surgery and prepare skin S9 fractions to measure the metabolic activity of Phase II enzymes. Preincubation of an excess of cofactors at 37 °C for fifteen minutes in the S9 before introduction of the testing probe, greatly increased the stability of the enzymes. Using this standardised assay, the rates of sulphation (SULT) and glucuronidation (UGT) of 7-hydroxycoumarin, methylation (COMT) of dopamine and N-acetylation (NAT) of procainamide were measured in the ng/mg protein/h (converted to ng/cm2/h) range in eighty-seven individuals. Glutathione conjugation (GST) of 1-chloro-2,4-dinitrobenzene was assessed in a smaller pool of fifty donors; the metabolic rate was much faster and measured over six minutes using a different methodology to express rates in μg/mg protein/min (converted to μg/cm2/min). A comprehensive statistical analysis of these results was carried out, separating donors by age, gender and metabolic rate measured.
    URI
    http://hdl.handle.net/10454/15866
    Version
    Accepted Manuscript
    Citation
    Spriggs S, Cubberley R, Loadman P et al (2018) A study of inter-individual variability in the Phase II metabolism of xenobiotics in human skin. Toxicology Letters. 292: 63-72.
    Link to publisher’s version
    https://doi.org/10.1016/j.toxlet.2018.04.011
    Type
    Article
    Collections
    Life Sciences Publications

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