Synthesis of stable isotope labelled steroid bis(sulfate) conjugates and their behaviour in collision induced dissociation experiments

Abstract

Selective incorporation of stable isotope labelled sulfate esters in steroidal systems affords internal standards and MS probes to investigate the fragmentation patterns of mono- and bis-conjugated derivatives in CID MS/MS experiments.

Energy-Resolved Fragmentation Aiding the Structure Elucidation of Steroid Biomarkers

“…The estimated threshold energies ( E 5% ) were derived from the energy ( E cm ) when I i = 5, i.e. the calculated energy ( E cm ) required to achieve 5% fragmentation, and were calculated as previously described. , …”

“…Energy-resolved CID has also been used to study the dissociation pathways of the Beta-2 agonist class of performance-enhancing drugs and to aid the structural characterization of N -glycans from their fragmentation patterns . Recently, these approaches have been extended to reveal characteristic differences between sets of related steroid monosulfates on the basis of estimated threshold energies ( E 5% ) . A common drawback in this energy-resolved CID analysis is the sensitivity of the measured energies to small changes in experimental and instrumental parameters .…”

“…37 Recently, these approaches have been extended to reveal characteristic differences between sets of related steroid monosulfates on the basis of estimated threshold energies (E 5% ). 38 A common drawback in this energy-resolved CID analysis is the sensitivity of the measured energies to small changes in experimental and instrumental parameters. 39 This variability suggests the need for a reference set of sulfated molecules to improve the reliability of experimentally measured threshold energies when used on different systems.…”

“…the calculated energy (E cm ) required to achieve 5% fragmentation, and were calculated as previously described. 35,38 ■ RESULTS AND DISCUSSION A Novel Metabolite of Testosterone Propionate in the Equine. In a recent study aimed at the untargeted profiling of equine urinary sulfate metabolites after testosterone propionate administration, 41 two putative steroid sulfate metabolites with the same exact mass (m/z 369.1739) were discovered.…”

Annual banned‐substance review—Analytical approaches in human sports drug testing 2021/2022–

“…By means of selective derivatization, the presence of 3‐hyroxymethyl‐ and 17‐hyroxymethyl‐conjugated M3 was corroborated, suggesting another simple and straightforward option for the sensitive detection of DHCMT misuse in sport. Metabolic patterns concerning both phase I and phase II biotransformations can be of particular interest and added value in anti‐doping detection strategies and case management, justifying the overall increasing interest in synthesizing and analyzing intact phase II AAS metabolites 106 . Although being commonly formulated as oral AASs, other routes of administration or exposure to DCHMT are conceivable as presented by Gessner et al, who investigated the resorption and elimination of DHCMT and other AASs after dermal application 26 .…”