Drug Testing and Analysis

This study showed an increase of a specific circulating miRNA, miR-144, in plasma samples after a single C.E.R.A injection. Moreover, this increase indicated that the miRNA method might be use for long term detection.

The detection of zeranol in urine samples might result from mycotoxin-contaminated cereal products. A differentiation of an illicit use as anabolic agent from an unintended ingestion of mycotoxin-colonized food is supported by the analysis of indicative metabolites and relative abundances.

All data from this excretion study and the reference data from our routine and research were measured with WADA approved Kit 1 and Kit 2. The figures and tebles show the statistical results of the concentrations of GH isoforms and their ratios, which support the time window of detection after administration of rhGH preparation in this excretion study.

This report describes the detection of an unknown truncated or modified form of FGF-1 in an injection vial confiscated by the German customs. The identification of the protein was accomplished by different proteomic approaches including gel electrophoresis, in-gel proteolysis and nano-liquid chromatography high-resolution/high-accuracy Orbitrap mass spectrometry.

Intravenous administration studies of epoetin alfa, epoetin beta and the new biosimilar epoetin kappa were performed to test the applicability of the two WADA-approved methods (IEF-, SDS-PAGE). The isoform bands of epoetin kappa expanded more widely towards the basic area in the IEF-PAGE and epoetin kappa contains isoforms with a higher molecular mass than those of other epoetins. The intravenous injections of epoetins showed shorter clearance times and the detection window of SDS-PAGE is longer than that of IEF-PAGE.

Dried blood spot sampling in doping controls provides the advantageous potential to obtain information of circulating drugs in pre- or post-composition testing with minimal invasive sample collection and superior transfer/ storage conditions.

The content and δ ¹³C_VPDB values of testosterone were determined for black marked testosterone products collected in Austria. For a significant amount of the products, the declared ingredients differed from the actual content. More than half of the preparations were found to have δ ¹³C_VPDB values of testosterone in the range reported for endogenous urinary steroids.

An assay for the determination of salbutamol and its glucuronide in human urine by means of direct injection utilizing liquid chromatography tandem mass spectrometry is presented and the excretion of small amounts of salbutamol glucuronide in human urine following the administration of salbutamol was proven.

The usefulness of four testosterone metabolites released after alkaline treatment has been evaluated for the detection of the misuse of single doses of dihydrotestosterone (DHT) gel, oral dehydroepiandrosterone (DHEA) and testosterone gel. Whereas limited efficacy was observed for DHT gel, the use of these metabolites allowed for increasing the retrospectivity in oral DHEA and testosterone gel administration.

A new multi-target approach based on (LC-(ESI)-MS/MS) is presented to screen for various classes of prohibited substances using direct injection of urine specimens. The assay enables the detection of classic groups of drugs, e.g. diuretics, beta2-agonists, stimulants and narcotics at concentration levels far below the required limits. Additionally, more challenging and various new target compounds could be implemented, e.g. stimulant conjugates, the plasma volume expanders dextran and hydroxyethyl starch, the selective androgen receptor modulator andarine, a metabolite of growth hormone releasing peptide (GHRP-2), 5-amino-4-imidazolecarboxyamide ribonucleoside (AICAR) and conjugates of di(2-ethylhexyl) phthalate (DEHP) metabolites indicating for illicit blood-transfusion are analysed.

The ethanol consumption marker ethylglucuronide was found to be a good marker for a potential alcohol induced alteration of steroid concentration in urine.

Comprehensive two dimensional gas chromatography coupled to fast quadrupole mass spectrometry (GC×GC-qMS) was evaluated for analysis of urinary steroids using electron ionization (EI) and positive chemical ionization with NH₃ (PCI-NH₃). PCI-NH₃ yielded highly specific mass spectra with characteristic pseudomolecular ions for steroid acetates which complement EI spectra for structural characterization.

Low molecular weight luteinizing hormone (LMWLH) receptor agonists could be of interest as a potential doping substance for athletes. Precursor ion monitoring liquid chromatography tandem mass spectrometry method was proposed to detect a broad range of potential LMWLH receptor agonists. The method was tested against a selection of urine samples to ascertain potential problems with background analytes interfering with the compounds of interests. The two available LMWLH receptor agonists could be detected at concentrations of 100 ng/mL in urine samples.

A number of supplements are now available which are sold as fat burners or pre-workout boosters and contain stimulants which are banned in sport. Many contain methylhexaneamine under one of many pseudonyms including Geranamine, geranium oil or extract, or a number of chemical names such as 1,3-dimethylpentylamine.

This communication shows that geranium oils do not contain methylhexaneamine and that products labeled as containing geranium oil but which contain methylhexaneamine can only arise from addition of synthetic material.

Glycerol was administered and hydration status and plasma and urinary glycerol concentrations were measured. Plasma volume increased but the effects on haemoglobin and haematocrit were rather small. Urinary glycerol concentrations were significantly higher than in placebo until 16.9 ± 1.0 h post administration.

A profound structural difference between human endogenous EPO and literally all recombinant erythropoietins was discovered and visualized by SDS-PAGE and sequential exoglycosidase digestion. According to these data all recombinant EPO pharmaceuticals currently produced worldwide significantly differ from human endogenous epoetins. Aside from its applicability in sports drug testing the discovery might prove useful for the production of recombinant epoetins with closer structural identity to human EPO.

Intra-individual variability of urinary di(2-ethylhexyl) phthalate (DEHP) metabolites were tested among seven volunteers during one week to investigate the possibility of increased DEHP exposure via non-medical routes. Quantification of the metabolites was accomplished after enzymatic hydrolysis of urinary glucuronide conjugates and direct injection using isotope-dilution liquid-chromatography/tandem mass spectrometry.

This study presents a screening procedure for the detection of saccharides and polyalcohols in human urine in the framework of doping control analysis. The proposed method was set up and validated to detect the abuse of dextran, hydroxyethyl starch and mannitol and involves only one enzymatic hydrolysis step and the direct injection into a LC-MS/MS system. The method could be used also as HES and dextran confirmation method avoiding the need for the complex confirmation procedures used until now.