Determination of selected endogenous anabolic androgenic steroids and ratios in urine by ultra high performance liquid chromatography tandem mass spectrometry and isotope pattern deconvolution

Abstract

8An isotope dilution mass spectrometry (IDMS) method for the determination of selected 9 endogenous anabolic androgenic steroids (EAAS) in urine by UHPLC-MS/MS has been 10 developed using the isotope pattern deconvolution (IPD) mathematical tool. The method has 11 been successfully validated for testosterone, epitestosterone, androsterone and 12 etiocholanolone, employing their respective deuterated analogs using two certified reference 13 materials (CRM). Accuracy was evaluated as recovery of the certified values and ranged from 14 75% to 108%. Precision was assessed in intraday (n=5) and interday (n=4) experiments, with 15RSDs below 5% and 10% respectively. The method was also found suitable for real urine 16 samples, with limits of detection (LOD) and quantification (LOQ) below the normal urinary 17 levels. The developed method meets the requirements established by the World Anti-Doping

Formation of multimeric steroid metal adducts and implications for isomer mixture separation by traveling wave ion mobility spectrometry

“…The extracted ATDs for testosterone and its isomers/isobars are illustrated in Figure 3. ultra-high performance LC coupled to tandem MS. 34 As fully combined solutions, the same trends were observed in the ATDs, whereby lithiated dimers were best suited for separation between epitestosterone/testosterone from DHEA, while sodiated dimers were preferred for distinguishing between epitestosterone and testosterone.…”

“…Epitestosterone and testosterone were partially resolved as sodiated dimers ( R S = 1.30), but only barely distinguishable as lithiated dimers ( R S = 0.42). Compared with LC methods, Pitarch‐Motellon et al achieved baseline separation of epitestosterone and testosterone in approximately 7 minutes with ultra‐high performance LC coupled to tandem MS . As fully combined solutions, the same trends were observed in the ATDs, whereby lithiated dimers were best suited for separation between epitestosterone/testosterone from DHEA, while sodiated dimers were preferred for distinguishing between epitestosterone and testosterone.…”

From fundamentals in calibration to modern methodologies: A tutorial for small molecules quantification in liquid chromatography–mass spectrometry bioanalysis

“…This result agrees with the high metrological quality of analytical results provided by isotope dilution mass spectrometry determination. The IPD approach was later validated with a larger pattern of steroids [153]. Then, the quantification of testosterone was improved by operating low resolution selected reaction monitoring (i.e., reduced resolution of the first quadrupole up to 8 m/z to transmit the entire ion cluster of the SIL and the analytes).…”

Determination of Ketosteroids in Human Urine Using Dispersive Liquid-Liquid Microextraction and Ultra High-Performance Liquid Chromatography-High Resolution Mass Spectrometry

“…Various methods of the preparation of urine samples for analysis are used: liquid-liquid extraction [11][12][13], stir bar sorptive extraction [14], on-line intube solid-phase microextraction [15], automated solid-phase extraction [16], dispersive liquid-liquid microextraction (DLLME) [8], and others [17]. Among the listed methods, the most promising is DLLME because of its simplicity, high concentration factors, and compliance with the criteria of "green chemistry" [18].…”