Use of dried blood spots in doping control analysis of anabolic steroid esters

Self Cite

Injections of synthetic esters of testosterone are among the most common forms of testosterone application. In doping control, the detection of an intact ester of testosterone in blood gives unequivocal proof of the administration of exogenous testosterone. The aim of the current project was to investigate the detection window for injected testosterone esters as a mixed substance preparation and as a single substance preparation in serum and plasma. Furthermore, the suitability of different types of blood collection devices was evaluated. Collection tubes with stabilizing additives, as well as non-stabilized serum separation tubes, were tested. A clinical study with six participants was carried out, comprising a single intramuscular injection of either 1000 mg testosterone undecanoate (Nebido(®)) or a mixture of 30 mg testosterone propionate, 60 mg testosterone phenylpropionate, 60 mg testosterone isocaproate, and 100 mg testosterone decanoate (Sustanon(®)). Blood was collected throughout a testing period of 60 days. The applied analytical method for blood analysis included liquid-liquid extraction and preparation of oxime derivatives, prior to TLX-sample clean-up and liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection. All investigated testosterone esters could be detected in post-administration blood samples. The detection time depended on the type of ester administered. Furthermore, results from the study show that measured blood concentrations of especially short-chained testosterone esters are influenced by the type of blood collection device applied. The testosterone ester detection window, however, was comparable.

Section: Introductionmentioning

confidence: 99%

Detection of testosterone esters in blood

Forsdahl

Erceg

Geisendorfer

et al. 2015

Self Cite

“…Among these materials, hair, sweat, saliva, urine and, currently, DBSs are some of these alternatives matrices that can be used in drug testing. [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] To verify that variations in FN1 concentrations could be detected in one DBS and urine after an rhGH administration, a pilot study with volunteers treated with a low dose of rhGH was performed. [38] Urine and DBSs were selected as potential matrices to be tested for the potential FN1 detection because they have provided promising results in the detection of a wide range of doping substances.…”

Section: Discussionmentioning

confidence: 99%

“…[2] Largely responsible for the presence of FN1 in plasma is its synthesis by hepatocytes although a smaller contribution by macrophages, lymphocytes, platelets, and endothelial cells is produced. [18] A steadily growing list of analytes related to doping substances may be measured from DBSs such as testosterone and others anabolic steroid esters, [19][20][21] haemoglobin, [22,23] pegylated erythropoietin-mimetic peptide, [24] growth hormone (GH), [25,26] IGF-I and IGF binding proteins, [27][28][29] soluble transferring receptors, [30] ferritin, [31] insulin, [32] opioids, [33] and ephedrine. [4][5][6][7] Several studies have shown FN1 as a potential genetic and protein biomarker in blood samples (peripheral blood lymphocytes (PBL), serum, and plasma) for the detection of insulin-like growth factor 1 (IGF-1) and human recombinant growth hormone (rhGH) abuse in sport.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of fibronectin 1 in one dried blood spot and in urine after rhGH treatment

Ferro

Ventura

Pérez‐Mañá

et al. 2016

Since the appearance of recombinant human growth hormone (rhGH) in the 1980s, its expansion and acquisition through the black market has increased, so the detection of its abuse continues to be a challenge. New biomarkers that are more reliable and sensitive, allowing a larger detection window, are still needed. In this line, Fibronectin 1 (FN1) has been proposed as a potential genetic and protein biomarker of rhGH abuse in peripheral blood lymphocytes, serum, and plasma. However, logistic problems associated with current blood collection in sports drug testing point towards potential new alternative matrices that could be good candidates to be evaluated. Results obtained in this study showed high ELISA FN1 levels in one dried blood spot and in urine samples in ten healthy male volunteers treated with rhGH. Results showed that especially dried blood spots appear as a potential good matrix to detect rhGH abuse by means of FN1 biomarker. Copyright © 2016 John Wiley & Sons, Ltd.

“…[23] However, official doping control testing for low-molecular-weight substances such as stimulants has not been introduced. [35][36][37][38][39] In this study, blood analyses for ephedrine and methylephedrine were conducted using DBS cards. In recent years, dried blood spot (DBS) testing, which exhibits advantages in storage, transportation, and invasiveness, has been employed in pharmaceutical sciences [24][25][26][27][28][29] and forensic sciences.…”

Section: Introductionmentioning

confidence: 99%

Comparison of urine analysis and dried blood spot analysis for the detection of ephedrine and methylephedrine in doping control

Kojima¹,

Nishitani²,

Sato³

et al. 2015

When the misuse of stimulants is determined in doping control tests conducted during the in-competition period, athletes are asked to account for the violation of the rules. This study was designed to evaluate whether the urinary threshold values (10 µg/mL) for ephedrine and methylephedrine set by the World Anti-Doping Agency (WADA) can be exceeded after the oral administration of each substance (25 mg). In addition, the study describes the validity of a liquid chromatography-tandem mass spectrometric method using dried blood spot testing to detect ephedrine and methylephedrine by comparing it to a quantitative laboratory urine assay. After administration of ephedrine, the urinary concentration of ephedrine did not exceed the threshold at 4-10 h in two subjects, whereas the threshold was exceeded in both the subjects at 12 h after administration. For methylephedrine, the urinary concentrations of all the subjects failed to reach the threshold for up to 10 h after administration. The concentrations reached the threshold at 12-24 h after administration in some volunteers. In contrast, the blood concentrations of ephedrine and methylephedrine reached their maximum levels at 2-8 h after administration. The blood concentrations showed a low inter-individual variability, and the results suggested that the urinary excretion of ephedrine and methylephedrine can be strongly affected by urine pH and/or urine volume. These facts suggest that urinary concentrations cannot reflect the psychoactive level of ephedrines in circulation. Thus, dried blood analysis might be suitable for the adequate detection of stimulants during in-competition testing.