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Stability and detectability of testosterone esters in Dried Blood Spots after intramuscular injections

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Solheim, Sara Amalie ; Levernaes, Maren Christin Stillesby ; Mørkeberg, Jakob ; Juul, Anders ; Upners, Emmie N ; Nordsborg, Nikolai Baastrup ; Dehnes, Yvette. / Stability and detectability of testosterone esters in Dried Blood Spots after intramuscular injections. I: Drug Testing and Analysis. 2021.

Bibtex

@article{fe55e11c02954860915f5bbc8f275946,
title = "Stability and detectability of testosterone esters in Dried Blood Spots after intramuscular injections",
abstract = "While misuse of testosterone esters is widespread in elite and recreational sports, direct detection of intact testosterone esters in doping control samples is hampered by the rapid hydrolysis by esterases present in the blood. With dried blood spot (DBS) as sample matrix, continued degradation of the esters is avoided due to inactivation of the hydrolase enzymes in dried blood. Here, we have developed the method further for detection of testosterone esters in DBS with focus on robustness and applicability in doping control. To demonstrate the method's feasibility, DBS samples from men receiving two intramuscular injections of Sustanon{\textregistered} 250 (n = 9) or placebo (n = 10) were collected, transported, and stored prior to analysis, to mimic a doping control scenario. The presented nanoLC-HRMS/MS method appeared reliable and suitable for direct detection of four testosterone esters (testosterone decanoate, isocaproate, phenylpropionate, and propionate) after extraction from DBS. Sustanon{\textregistered} was detected in all subjects for at least 5 days, with detection window up to 14 days for three of the esters. Evaluation of analyte stability showed that while storage at room temperature is tolerated well for a few days, testosterone esters are highly stable (>18 months) in DBS when stored in frozen conditions. Collectively, these findings demonstrate the applicability of DBS sampling in doping control for detection of steroid esters. The fast collection and reduced shipment costs of DBS compared with urine and standard blood samples, respectively, will allow more frequent and/or large-scale testing to increase detection and deterrence.",
keywords = "anabolic steroid esters, doping control analysis, dried blood spots (DBS), mass spectrometry",
author = "Solheim, {Sara Amalie} and Levernaes, {Maren Christin Stillesby} and Jakob M{\o}rkeberg and Anders Juul and Upners, {Emmie N} and Nordsborg, {Nikolai Baastrup} and Yvette Dehnes",
note = "This article is protected by copyright. All rights reserved.",
year = "2021",
month = mar,
day = "17",
doi = "10.1002/dta.3030",
language = "English",
journal = "Drug Testing and Analysis",
issn = "1942-7603",
publisher = "JohnWiley & Sons Ltd",

}

RIS

TY - JOUR

T1 - Stability and detectability of testosterone esters in Dried Blood Spots after intramuscular injections

AU - Solheim, Sara Amalie

AU - Levernaes, Maren Christin Stillesby

AU - Mørkeberg, Jakob

AU - Juul, Anders

AU - Upners, Emmie N

AU - Nordsborg, Nikolai Baastrup

AU - Dehnes, Yvette

N1 - This article is protected by copyright. All rights reserved.

PY - 2021/3/17

Y1 - 2021/3/17

N2 - While misuse of testosterone esters is widespread in elite and recreational sports, direct detection of intact testosterone esters in doping control samples is hampered by the rapid hydrolysis by esterases present in the blood. With dried blood spot (DBS) as sample matrix, continued degradation of the esters is avoided due to inactivation of the hydrolase enzymes in dried blood. Here, we have developed the method further for detection of testosterone esters in DBS with focus on robustness and applicability in doping control. To demonstrate the method's feasibility, DBS samples from men receiving two intramuscular injections of Sustanon® 250 (n = 9) or placebo (n = 10) were collected, transported, and stored prior to analysis, to mimic a doping control scenario. The presented nanoLC-HRMS/MS method appeared reliable and suitable for direct detection of four testosterone esters (testosterone decanoate, isocaproate, phenylpropionate, and propionate) after extraction from DBS. Sustanon® was detected in all subjects for at least 5 days, with detection window up to 14 days for three of the esters. Evaluation of analyte stability showed that while storage at room temperature is tolerated well for a few days, testosterone esters are highly stable (>18 months) in DBS when stored in frozen conditions. Collectively, these findings demonstrate the applicability of DBS sampling in doping control for detection of steroid esters. The fast collection and reduced shipment costs of DBS compared with urine and standard blood samples, respectively, will allow more frequent and/or large-scale testing to increase detection and deterrence.

AB - While misuse of testosterone esters is widespread in elite and recreational sports, direct detection of intact testosterone esters in doping control samples is hampered by the rapid hydrolysis by esterases present in the blood. With dried blood spot (DBS) as sample matrix, continued degradation of the esters is avoided due to inactivation of the hydrolase enzymes in dried blood. Here, we have developed the method further for detection of testosterone esters in DBS with focus on robustness and applicability in doping control. To demonstrate the method's feasibility, DBS samples from men receiving two intramuscular injections of Sustanon® 250 (n = 9) or placebo (n = 10) were collected, transported, and stored prior to analysis, to mimic a doping control scenario. The presented nanoLC-HRMS/MS method appeared reliable and suitable for direct detection of four testosterone esters (testosterone decanoate, isocaproate, phenylpropionate, and propionate) after extraction from DBS. Sustanon® was detected in all subjects for at least 5 days, with detection window up to 14 days for three of the esters. Evaluation of analyte stability showed that while storage at room temperature is tolerated well for a few days, testosterone esters are highly stable (>18 months) in DBS when stored in frozen conditions. Collectively, these findings demonstrate the applicability of DBS sampling in doping control for detection of steroid esters. The fast collection and reduced shipment costs of DBS compared with urine and standard blood samples, respectively, will allow more frequent and/or large-scale testing to increase detection and deterrence.

KW - anabolic steroid esters

KW - doping control analysis

KW - dried blood spots (DBS)

KW - mass spectrometry

UR - http://www.scopus.com/inward/record.url?scp=85110050578&partnerID=8YFLogxK

U2 - 10.1002/dta.3030

DO - 10.1002/dta.3030

M3 - Journal article

C2 - 33733610

JO - Drug Testing and Analysis

JF - Drug Testing and Analysis

SN - 1942-7603

ER -

ID: 66477950