Three Hydrolysis Methods for 17-Ketosteroid Sulfates Compared by Colorimetric and Gas—Liquid Chromatographic Analyses

Muehlbaecher, C. A.; Smith, Everett L.

doi:10.1093/clinchem/16.3.158

Cited by 9 publications

(2 citation statements)

References 6 publications

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“…Traditionally, chemical hydrolysis was achieved using hot acid with cleavage of the conjugate being strongly influenced by the choice of acid (hydrochloric or sulfuric), acid molarity, temperature and duration of the reaction [32] . More recently, chemical hydrolysis has been carried out by solvolysis and when compared to hot acid hydrolysis, leads to superior steroid recovery [33] . Solvolysis can utilise ethyl acetate in acidic conditions and heating for anything between 1 h at 55 °C [26] , and 24–48 h at 37 °C [18,34] .…”

Section: Deconjugation Approachesmentioning

confidence: 99%

Analysis of conjugated steroid androgens: Deconjugation, derivatisation and associated issues

Gomes

Meredith

Snape

et al. 2009

Journal of Pharmaceutical and Biomedical Analysis

109

104

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Gas chromatography/mass spectrometry (GC/MS) is the preferred technique for the detection of urinary steroid androgens for drug testing in athletics. Excreted in either the glucuronide or sulfated conjugated form, steroids must first undergo deconjugation followed by derivatisation to render them suitable for GC analysis. Discussed herein are the deconjugation and the derivatisation preparative options. The analytical challenges surrounding these preparatory approaches, in particular the inability to cleave the sulfate moiety have led to a focus on testing protocols that reply on glucuronide conjugates. Other approaches which alleviate the need for deconjugation and derivatisation are also highlighted.

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Section: Deconjugation Approachesmentioning

confidence: 99%

Analysis of conjugated steroid androgens: Deconjugation, derivatisation and associated issues

Gomes

Meredith

Snape

et al. 2009

Journal of Pharmaceutical and Biomedical Analysis

109

104

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“…Traditionally acidic hydrolysis at elevated temperatures was used for deconjugation of sterol sulfates. But the drastic conditions that are required for this hydrolysis including high amounts of mineral acid and refluxing, can lead to degradation or transformation of some sterols [51][52][53]. In turn, solvolysis under mild conditions is preferred and can be achieved by extracting the sterol sulfates from an acidified (with sulfuric acid) aqueous sample with ethyl acetate and storing this moist organic phase for 24 h at 39 • C [54] or with trimethylchlorosilane in methanol (methanolysis) [30].…”

Section: Chemical Cleavage Of Sterol Sulfatesmentioning

confidence: 99%

Comparison of Strategies for the Determination of Sterol Sulfates via GC-MS Leading to a Novel Deconjugation-Derivatization Protocol

et al. 2019

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Sulfoconjugates of sterols play important roles as neurosteroids, neurotransmitters, and ion channel ligands in health and disease. In most cases, sterol conjugate analysis is performed with liquid chromatography-mass spectrometry. This is a valuable tool for routine analytics with the advantage of direct sterol sulfates analysis without previous cleavage and/or derivatization. The complementary technique gas chromatography-mass spectrometry (GC-MS) is a preeminent discovery tool in the field of sterolomics, but the analysis of sterol sulfates is hampered by mandatory deconjugation and derivatization. Despite the difficulties in sample workup, GC-MS is an indispensable tool for untargeted analysis and steroid profiling. There are no general sample preparation protocols for sterol sulfate analysis using GC-MS. In this study we present a reinvestigation and evaluation of different deconjugation and derivatization procedures with a set of representative sterol sulfates. The advantages and disadvantages of trimethylsilyl (TMS), methyloxime-trimethylsilyl (MO-TMS), and trifluoroacetyl (TFA) derivatives were examined. Different published procedures of sterol sulfate deconjugation, including enzymatic and chemical cleavage, were reinvestigated and examined for diverse sterol sulfates. Finally, we present a new protocol for the chemical cleavage of sterol sulfates, allowing for simultaneous deconjugation and derivatization, simplifying GC-MS based sterol sulfate analysis.

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Zimmermann Reaction

2010

Comprehensive Organic Name Reactions and Reagents

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The reaction between an enolate carbanion and m ‐dinitrobenzene ( m ‐DNB) has been reported by Janovsky and Erb and it is Zimmermann who initially applied the specific reaction between m ‐dinitrobenzene and 17‐ oxo ‐steroids (or 17‐ keto ‐steroids) in the presence of an alkali to the field of ketosteroid analysis. This kind of reaction is generally known as the Zimmermann reaction. Due to the specific color reaction for 17‐ oxo ‐steroids, the Zimmermann reaction has been routinely used for the various clinical testings. Besides the interference from carbonyl compounds, several limitations of this reaction have been discussed.

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Three Hydrolysis Methods for 17-Ketosteroid Sulfates Compared by Colorimetric and Gas—Liquid Chromatographic Analyses

Cited by 9 publications

References 6 publications

Analysis of conjugated steroid androgens: Deconjugation, derivatisation and associated issues

Analysis of conjugated steroid androgens: Deconjugation, derivatisation and associated issues

Comparison of Strategies for the Determination of Sterol Sulfates via GC-MS Leading to a Novel Deconjugation-Derivatization Protocol

Zimmermann Reaction

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