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Selective reactions in the analysis and characterization of steroids by gas chromatography-mass spectrometry

  • Symposium: Sterol Analysis Held at the AOCS Annual Meeting in New York, NY, April 29, 1980
  • Published:
Lipids

Abstract

Gas chromatography-mass spectrometry (GC-MS) is a technique especially suitable for the analysis and characterization of steroids, and its power has been extensively demonstrated. The efficacy of GC-MS is limited, nevertheless, by the fact that steroid mixtures—whether of natural origin only, or augmented by synthetic analogs—often contain similar components that are poorly distinguished. The fortuitous overlap of gas chromatographic peaks from disparate compounds also impairs the definition of retention data. Controlled modification of the sample by means of selective reactions is therefore a valuable adjunct to the application of GC-MS. Two examples are discussed: (a) the enzyme cholesterol oxidase, isolated from various microorganisms, catalyzes the oxidation of many 3β-hydroxy-5-enes (with concomitant isomerization) to 4-en-3-ones; 3β-hydroxy-5α-steroids are also oxidized to the corresponding 3-ones, but other steroids (3α-hydroxy- or 5β-isomers, etc.) are unaffected. The mild conditions required (pH 7, 30 C) are advantageous for the analysis of sensitive steroids, and the retention index increments, as well as the mass spectra of the ketones, are characteristic. The enzyme accepts as substrates a wide range of 3β-hydroxysteroids, tolerating oxygenation in ring B and even catalyzing the oxidation of 2-oxacholesterol to the expected lactone; and (b) steroids possessing 1,2-diol or 1,3-diol groupings include estriols, 2-hydroxyestrone, 20,22-dihydroxycholesterols, ecdysones, brassinolide and many corticosteroids. The selective formation of cyclic derivatives can provide several analytically useful features, such as convenient retention times, moderate mass increments (24 amu for a methaneboronate), distinctive mass spectra and usually abundant molecular ions. These are exemplified for 5-pregnene-3β-20,21-triols and for 20,22-dihydroxycholesterol as well as its enzymic oxidation product.

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Authors and Affiliations

  1. Department of Chemistry, University of Glasgow, G12 8QQ, Glasgow

    C. J. W. Brooks, W. J. Cole & H. B. McIntyre

  2. M.R.C. Toxicology Unit, SM5 4EF, Carshalton, Surrey, U.K.

    A. G. Smith

Authors
  1. C. J. W. Brooks
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  2. W. J. Cole
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  3. H. B. McIntyre
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  4. A. G. Smith
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Brooks, C.J.W., Cole, W.J., McIntyre, H.B. et al. Selective reactions in the analysis and characterization of steroids by gas chromatography-mass spectrometry. Lipids 15, 745–755 (1980). https://doi.org/10.1007/BF02534028

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  • DOI: https://doi.org/10.1007/BF02534028

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