Important considerations for the utilisation of methanolysis in steroid analysis

Viljanto, Marjaana; Pita, Catarina Horro; Scarth, James; Walker, Christopher J.; Kicman, Andrew T.; Parkin, Mark C.

doi:10.1002/dta.2402

Cited by 6 publications

(2 citation statements)

References 18 publications

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“…Chemical hydrolysis offers an alternative to enzymatic hydrolysis, involving either solvolysis (Hauser et al, 2008) or hot acid (typically HCl or H 2 SO 4 ) hydrolysis (Dumasia & Houghton, 1981; Konieczna et al, 2011; Pizzato et al, 2017). Many hydrolyses undergo methanolysis, a variation of solvolysis (Cooper et al, 2001; Tseng et al, 2006), in which the oxygen attached to C‐17 undergoes protonation during strong acidic conditions, utilizing acetyl chloride in methanol, resulting in the simultaneous separation of sulfate and glucuronide moieties (Viljanto et al, 2018). Chemical hydrolysis via ethyl acetate/methanol/sulfuric acid 80:20:0,12 (v/v) provided the best hydrolysis response of at least 50% across testosterone, epitestosterone, androsterone, etiocholanolone, 5‐androstene‐3β,17β‐diol, 5α‐androstane‐3β,17β‐diol, DHEA, epiandrosterone, 11‐ketoetiocholanolone and cholesterol for the cleaving of the sulfate moiety in comparison to enzymatic hydrolysis (Iannone et al, 2020).…”

Section: Sample Pretreatment and Extraction Methodsmentioning

confidence: 99%

A review of the analytical techniques for the detection of anabolic–androgenic steroids within biological matrices

Harries,

De Paoli,

Hall

et al. 2023

WIREs Forensic Science

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Anabolic–androgenic steroids (AASs) and other image and performance enhancing drugs (IPEDs) are controlled by governments and sport institutions such as the World Anti‐doping Agency (WADA). Although elite athletes and professional bodybuilders are the most visible AAS abusers, the introduction of the internet has increased the accessibility of AASs, with use being observed among recreational gym goers at increasing prevalence. Despite reported increase in use, routine analysis for these substances is uncommon, with many forensic laboratories opting to outsource AAS analysis. This review collates information regarding the extraction and analysis of AASs from various biological matrices with the considered purpose of providing a reference for the development of AAS methods to allow for routine detection by forensic laboratories.This article is categorized under: Toxicology > in Sport Toxicology > Analytical

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Section: Sample Pretreatment and Extraction Methodsmentioning

confidence: 99%

A review of the analytical techniques for the detection of anabolic–androgenic steroids within biological matrices

Harries,

De Paoli,

Hall

et al. 2023

WIREs Forensic Science

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“…Deuterated IS can potentially experience hydrogen-deuterium exchange ( Kwok et al, 2008 ; Viljanto et al, 2018 ), so we tested for possible alterations of 17β-E 2 -d4 caused by the derivatization procedure. We compared the mass spectra of 17β-E 2 -d4 directly from the stock solution, after sham derivatization (resuspension in buffer and acetone followed by incubation for 15 min at 60°C, without DMIS), or after derivatization (resuspension in buffer and DMIS in acetone followed by incubation for 15 min at 60°C).…”

Section: Methodsmentioning

confidence: 99%

Ultrasensitive Quantification of Multiple Estrogens in Songbird Blood and Microdissected Brain by LC-MS/MS

Jalabert

Shock

et al. 2022

eNeuro

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Neuroestrogens are synthesized within the brain and regulate social behavior, learning and memory, and cognition. In song sparrows, Melospiza melodia, 17β-estradiol (17β-E 2 ) promotes aggressive behavior, including during the non-breeding season when circulating steroid levels are low. Estrogens are challenging to measure because they are present at very low levels, and current techniques often lack the sensitivity required.Furthermore, current methods often focus on 17β-E 2 and disregard other estrogens.Here, we developed and validated a method to measure four estrogens (estrone, 17β-E 2 , 17α-estradiol, estriol) simultaneously in microdissected songbird brain, with high specificity, sensitivity, accuracy, and precision. We used liquid chromatography tandem mass spectrometry (LC-MS/MS), and to improve sensitivity, we derivatized estrogens using 1,2-dimethylimidazole-5-sulfonyl-chloride (DMIS). The straightforward protocol improved sensitivity by 10-fold for some analytes. There is substantial regional variation in neuroestrogen levels in brain areas that regulate social behavior in male song sparrows. For example, the auditory area NCM, which has high aromatase levels, has the highest estrone and 17β-E 2 levels. In contrast, estrogen levels in blood are very low. Estrogen levels in both brain and circulation are lower in the non-breeding season than in the breeding season. This technique will be useful for estrogen measurement in songbirds and potentially other animal models.

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Analytical advances in horseracing medication and doping control from 2018 to 2023

Gray,

Lubbock,

Love

et al. 2024

Drug Testing and Analysis

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The analytical approaches taken by laboratories to implement robust and efficient regulation of horseracing medication and doping control are complex and constantly evolving. Each laboratory's approach will be dictated by differences in regulatory, economic and scientific drivers specific to their local environment. However, in general, laboratories will all be undertaking developments and improvements to their screening strategies in order to meet new and emerging threats as well as provide improved service to their customers. In this paper, the published analytical advances in horseracing medication and doping control since the 22nd International Conference of Racing Analysts and Veterinarians will be reviewed. Due to the unprecedented impact of COVID‐19 on the worldwide economy, the normal 2‐year period of this review was extended to over 5 years. As such, there was considerable ground to cover, resulting in an increase in the number of relevant publications included from 107 to 307. Major trends in publications will be summarised and possible future directions highlighted. This will cover developments in the detection of ‘small’ and ‘large’ molecule drugs, sample preparation procedures and the use of alternative matrices, instrumental advances/applications, drug metabolism and pharmacokinetics, the detection and prevalence of ‘endogenous' compounds and biomarker and OMICs approaches. Particular emphasis will be given to research into the potential threat of gene doping, which is a significant area of new and continued research for many laboratories. Furthermore, developments in analytical instrumentation relevant to equine medication and doping control will be discussed.

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Important considerations for the utilisation of methanolysis in steroid analysis

Cited by 6 publications

References 18 publications

A review of the analytical techniques for the detection of anabolic–androgenic steroids within biological matrices

A review of the analytical techniques for the detection of anabolic–androgenic steroids within biological matrices

Ultrasensitive Quantification of Multiple Estrogens in Songbird Blood and Microdissected Brain by LC-MS/MS

Analytical advances in horseracing medication and doping control from 2018 to 2023

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