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HPLC / MSN Analysis of Retinoids

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Book cover Retinoids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 652))

Abstract

This protocol describes a highly sensitive and selective method to quantify retinoids using normal-phase HPLC with online APCI MSN. The retinoids are key regulators of gene expression, retinol being oxidized via a retinaldehyde intermediate to retinoic acid (RA) which activates specific nuclear receptors, the signalling of which is turned off by oxidative inactivation of the ligand to 4-oxo-RA and other metabolites. Many of these retinoids are present only transiently at low concentrations in tissues and during analysis are labile to heat, light, and oxygen. HPLC with online APCI MSN provides a rapid technique to quantify these retinoids simultaneously. Techniques to extract the retinoids and prevent their degradation are described, with an emphasis on transcriptionally active RA. RA controls patterning of gene expression in the embryo, organizing embryonic morphology in the central nervous system. Similarly, a patterned distribution of RA controls function of the adult CNS, a tissue particularly difficult to analyse for RA because of its high lipid content. To understand how these patterns are organized in the brain and change over time, it is essential to determine the concentration of RA in small areas of tissues, and techniques of exquisite sensitivity are indispensable.

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Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 01655, Worcester, MA, USA

    James E. Evans

  2. Institute of Medical Sciences, School of Medical Sciences, Foresterhill, Aberdeen, Scotland

    Peter McCaffery

Authors
  1. James E. Evans
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  2. Peter McCaffery
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Editor information

Editors and Affiliations

  1. David Geffen School of Medicine, Dept. Physiology, University of California, Los Angeles, Charles Young Dr. S. 650, Los Angeles, 90095-1751, California, USA

    Hui Sun

  2. , Jules Stein Eye Institute, UCLA School of Medicine, 100 Stein Plaza, BH-667, Los Angeles, 90095, California, USA

    Gabriel H. Travis

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Evans, J.E., McCaffery, P. (2010). HPLC / MSN Analysis of Retinoids. In: Sun, H., Travis, G. (eds) Retinoids. Methods in Molecular Biology, vol 652. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-325-1_8

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  • DOI: https://doi.org/10.1007/978-1-60327-325-1_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-324-4

  • Online ISBN: 978-1-60327-325-1

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