Measuring urinary testosterone levels of the great apes—Problems with enzymatic hydrolysis using Helix pomatia juice

Hauser, Barbara; Schulz, D.; Boesch, Christophe; Deschner, Tobias

doi:10.1016/j.ygcen.2008.05.006

Cited by 30 publications

(26 citation statements)

References 34 publications

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“…However, no steroids with 3α-hydroxy-5-ene, 3α-hydroxy-5α or 5β-reduced, and 3-oxo-5α or 5β-reduced structure produced any by-products, which are accordance with a previous report (16). These results suggest that Helix pomatia also contains cholesterol oxidase in addition to 3β-hydroxysteroid oxidoreductase/3-oxosteroid-4,5-ene isomerase and 6-hydroxylase, 6-hydroxysteroid as additional enzymes (16)(17)(18)(20)(21)(22)(23)(24).…”

Section: Resultssupporting

confidence: 91%

“…These two actions suggest that they are caused by the presence of 3β-hydroxysteroid dehydrogenase/Δ 5-4 -ene steroid isomerase and 6-hydroxylase as additional enzymes (17,18,20) in the Helix pomatia extracts. The generation of 3-oxo-4-ene steroids might be converted by cholesterol oxidase because 3β-hydroxysteroid dehydrogenase does not require oxygen (16,17,(21)(22)(23)(24), but this has not been proven. The variability of the selectivity and reactivity of Helix pomatia is also affected by the reaction temperature, incubation time pH, and amount of enzyme added (14,(25)(26)(27).…”

Section: Introductionmentioning

confidence: 99%

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Systematic Error in Gas Chromatography-Mass Spectrometry–Based Quantification of Hydrolyzed Urinary Steroids

Moon

et al. 2010

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Gas chromatography-mass spectrometry-based metabolite profiling can lead to an understanding of various disease mechanisms as well as to identifying new diagnostic biomarkers by comparing the metabolites related in quantification. However, the unexpected transformation of urinary steroids during enzymatic hydrolysis with Helix pomatia could result in an underestimation or overestimation of their concentrations. A comparison of β-glucurondase extracted from Escherichia coli revealed 18 conversions of 84 steroids tested as an unexpected transformation under hydrolysis with β-glucuronidase/arylsulfatase extracted from Helix pomatia. In addition to the conversion of 3β-hydroxy-5-ene steroids into 3-oxo-4-ene steroids, which has been reported, the transformation of 3β-hydroxy-5α-reduced and 3β-hydroxy-5β-reduced steroids to 3-oxo-5α-reduced and 3-oxo-5β-reduced steroids, respectively, was newly observed. The formation of by-products was in proportion to the concentration of substrates becoming saturated against the enzyme. The substances belonging to these three steroid groups were undetectable at low concentrations, whereas the corresponding by-products were overestimated. These results indicate that the systematic error in the quantification of urinary steroids hydrolyzed with Helix pomatia can lead to a misreading of the clinical implications. All these hydrolysis procedures are suitable for study purposes, and the information can help prevent false evaluations of urinary steroids in clinical studies. Cancer Epidemiol Biomarkers Prev; 19(2); 388-97. ©2010 AACR.

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Section: Resultssupporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Systematic Error in Gas Chromatography-Mass Spectrometry–Based Quantification of Hydrolyzed Urinary Steroids

Moon

et al. 2010

Cancer Epidemiology, Biomarkers &Amp; Prevention

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“…Indeed, previous studies demonstrated that the use of Helix pomatia juice can produce misleading results due to the presence of other enzymes that are responsible for steroids conversion [48, 49] that could affect the correct determination of 17 α -T, AED, DHEA, DHT, and ETIO. This different approach was used for the secondary goal of performing multivariate analyses based on the urinary profile of endogenous steroids to develop a probabilistic model able to discriminate treated animals from nontreated ones.…”

Section: Resultsmentioning

confidence: 99%

Urinary Concentrations of Steroids in Bulls under Anabolic Treatment by Revalor-XS® Implant

Biancotto

Stella

Barrucci

et al. 2016

Journal of Analytical Methods in Chemistry

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Despite the European ban of using anabolics in food-producing animals, growth promoters might still be illegally used in the European Union. To control the food chain and guarantee consumers' health, there is a need of highly sensitive analytical methods for the identification of marker residues of such treatments. In the present study, a group of bulls (n = 16) received trenbolone acetate (200 mg) and estradiol (40 mg) by a commercial ear implant during a time range of 71 days, and a second group (n = 16) was kept for control. The aim of the research was to measure the residual urinary concentrations of the administered drugs (β-trenbolone and β-estradiol), their main metabolites (α-trenbolone and α-estradiol), and possible alterations of the urinary profile of other endogenous hormones metabolically related. The analytical method was based on liquid chromatography-tandem mass spectrometry. Results showed average urinary concentrations of α-trenbolone and α-estradiol during treatment in the range of (0.81 ÷ 2.1) ng mL−1 and (0.96 ÷ 4.4) ng mL−1, respectively, whereas β-trenbolone and β-estradiol exhibit urinary concentrations lower than 0.22 ng mL−1 in both cases. Data obtained from the urinary profiles of endogenous steroids indicate that they could be useful to indirectly detect the ongoing treatment.

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“…Shideler et al, 1994) or as the free form after cleavage of the conjugate using enzymatic hydrolysis or solvolysis followed by use of an assay designed to measure the unconjugated compound (Eastman et al, 1984;Ziegler et al, 1996). In case of enzymatic hydrolysis, application should be viewed with caution when using enzyme preparations from Helix pomatia since these contain enzymes other than glucuronidase/arylsuphatase which can result in hormone transformation (see Hauser et al, 2008).…”

Section: Type Of Urinary Assaysmentioning

confidence: 99%

Non-invasive monitoring of endocrine status in laboratory primates: methods, guidelines and applications

Heistermann

2010

Adv. Sci. Res.

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Abstract. During the past three decades, non-invasive methods for assessing physiological, in particular endocrine, status have revolutionized almost all areas of primatology, including behavioural ecology, reproductive biology, stress research, conservation and last but not least management of primates in captivity where the technology plays an integral role in assisting the husbandry, breeding and welfare of many species. Noninvasive endocrine methods make use of the fact that hormones circulating in blood are secreted into saliva or deposited in hair and are eliminated from the body via urinary and faecal excretion. The choice of which matrix to use for hormonal assessment depends on a range of factors, including the type of information required, the measurement techniques involved, species differences in hormone metabolism and route of excretion and the practicality of sample collection. However, although sample collection is usually relatively easy, analysing hormones from these non-invasively collected samples is not as easy as many people think, particularly not when dealing with a new species. In this respect, the importance of a careful validation of each technique is essential in order to generate meaningful and accurate results. This paper aims to provide an overview of the available non-invasive endocrine-based methodologies, their relative merits and their potential areas of application for assessing endocrine status in primates, with special reference to captive environments. In addition, general information is given about the most important aspects and caveats researchers have to be aware of when using these methodologies.

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Measuring urinary testosterone levels of the great apes—Problems with enzymatic hydrolysis using Helix pomatia juice

Cited by 30 publications

References 34 publications

Systematic Error in Gas Chromatography-Mass Spectrometry–Based Quantification of Hydrolyzed Urinary Steroids

Systematic Error in Gas Chromatography-Mass Spectrometry–Based Quantification of Hydrolyzed Urinary Steroids

Urinary Concentrations of Steroids in Bulls under Anabolic Treatment by Revalor-XS® Implant

Non-invasive monitoring of endocrine status in laboratory primates: methods, guidelines and applications

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