Searching for in silico predicted metabolites and designer modifications of (cortico)steroids in urine by high‐resolution liquid chromatography/time‐of‐flight mass spectrometry

Abstract

Glucocorticosteroids are a restricted class of substances and appear on the 'in-competition' prohibited list of the World Anti-Doping Agency (WADA). Analysis of glucocorticosteroids is complicated since they show significant phase 1 and 2 metabolism in the human body and are excreted into urine in concentrations in the microg/L range. Full scan, high-resolution time-of-flight mass spectrometry analysis generates information on all ionisable components in urine, including known and unknown metabolites of steroids and even designer modifications of anabolic steroids. However, evaluation of the data obtained can be difficult and time-consuming because of the need to differentiate between endogenous components and compounds of interest. MetaboLynx, a spectral and chromatographic search program, was modified for the determination of in silico predicted metabolites of glucocorticosteroids and designer modifications of anabolic steroids in human urine. Spiked urine samples were successfully screened for known components in a targeted approach and for unknown species in a non-targeted approach using data filtering to limit potential false-positives. A simplified combined approach of targeted and untargeted screening was used for the detection of metabolites and designer modifications of existing compounds. This approach proved successful and showed its strength in the detection of tetrahydrogestrinone (THG), a designer modification of gestrinone. THG was positively detected in a spiked urine sample and correctly identified as a twofold hydrogenation of gestrinone. The developed screening method can easily be adapted to specific needs and it is envisaged that a similar approach would be amendable to the discovery of metabolites or designer modifications of other compounds of interest.

Fast and automated characterization of major constituents in rat biofluid after oral administration of Abelmoschus manihot extract using ultra‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry and MetaboLynx

“…In this paper, we describe a strategy using ultra‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry (UPLC/QTOFMS) with automated data analysis (MetaboLynx™) for fast analysis of the metabolic profile of Abelmoschus manihot . Although there have been reports of the use of the UPLC/QTOF/MetaboLynx approach to study the metabolic profiles of drugs and natural products,30, 36 it is still worthwhile to use this approach to study the metabolic profile of Traditional Chinese Medicines.…”

Current Status and Bioanalytical Challenges in The Detection of Unknown Anabolic Androgenic Steroids in Doping Control Analysis

“…Hence, the development of full acquisition methods based on HR instru ments is a promising alternative, not only for the open detection of AAS but also for the compre hensive target screening in doping control ana lysis (Table 2) [50][51][52][53][54][55][56][57][58][59][60][61][62][63][64][65][66]. Their modus operandi is the acquisition of the fullscan raw data and, therefore, the detection of all compounds ion ized by the source.…”

Exposure science in an age of rapidly changing climate: challenges and opportunities

“… 71 Instruments coupling gas chromatography or liquid chromatography with high-resolution mass spectrometry are most commonly used for NTA. These methods, and similar “suspect screening” methods, have been successfully applied to characterize chemical unknowns in environmental media including water, 72 lake sediment, 73 dust, 74 food, 75 and products; 76 and in biological media including animal tissue, 77 and human blood, 78 urine, 79 and teeth. 80 …”