An improved gas chromatography screening method for doping substances using triple quadrupole mass spectrometry, with an emphasis on quality assurance

Abstract

A GC‐QqQ‐MS method was developed for the detection of over 150 compounds from different classes (steroids, narcotics, stimulants, β‐blockers, β‐2‐agonists and hormone antagonists) in a qualitative way. In the quantitative part, the traditional steroid profile with the most important endogenous steroids is expanded with six minor metabolites, which further improves the detection and identification of endogenous steroid abuse. In addition to these, norandrosterone, salbutamol and the major metabolite of cannabis are also quantified. Methods developed for anti‐doping purposes should be subjected to the highest level of quality. Here, the addition of a combination of (deuterated) internal standards allows for an accurate quality control of every single step of the methodology: hydrolysis efficiency, derivatization efficiency and microbiological degradation are monitored in every single sample. Additionally, special attention is paid to the relationships between parameters indicating degradation by micro‐organisms and the reliability of the steroid profile. The impact of the degradation is studied by evaluation of the quantities and percentages of 5α‐androstane‐3,17‐dione and 5β‐androstane‐3,17‐dione. The concept of measurement uncertainty was introduced for the evaluation of relative abundances of mass‐to‐charge ratios and the obtained ranges were compared with the World Anti‐Doping Agency regulations on tolerance windows for relative ion intensities. The results indicate that the approaches are similar. Copyright © 2012 John Wiley & Sons, Ltd.

Discordant genotyping results using DNA isolated from anti‐doping control urine samples

“…[16,17] The limit of quantification was 0.5 ng/mL for both testosterone and epitestosterone. [16,17] The limit of quantification was 0.5 ng/mL for both testosterone and epitestosterone.…”

Annual banned‐substance review: analytical approaches in human sports drug testing

“…[9,10] In line with this trend, Musenga and Cowan reported on a fast screening/high throughput doping control assay in 2013. Employing gas chromatographytriple quadrupole (QqQ) tandem mass spectrometry (GC-MS/MS), a complementary methodology was presented in 2012, [12] allowing for the qualitative and partly quantitative identification of 173 analytes including representatives of anabolic agents, β 2 -agonists, hormone and metabolic modulators, masking agents, stimulants, narcotics, β-blockers, and cannabinoids. The eluate (3 mL of 3% ammonium hydroxide in methanol/acetonitrile, 1:1, v/v) was evaporated to dryness, and the residue reconstituted in 100 μL of aqueous formic acid (0.3%) containing 5% acetonitrile for analysis on an ultra high pressure liquid chromatography (UHPLC) system interfaced to an orbitrap mass spectrometer.…”

Improved sensitivity by use of gas chromatography—positive chemical ionization triple quadrupole mass spectrometry for the analysis of drug related substances

“…In all these cases the most abundant form is monitored. As mentioned in our previous publications, the derivatization efficiency and reproducibility is controlled by monitoring the monoTMS/bisTMS androsterone ratio in every sample [17,20].…”

“…Acetonitrile and methyl tert-butyl ether were from Biosolve (Valkenswaard, The Netherlands) and ammoniumiodide (NH 4 I) and dodecane were from Sigma Aldrich (Bornem, Belgium). The origin of all the reference standards is described in detail in our previous work [17,20].…”