Sports drug testing using complementary matrices: Advantages and limitations
Graphical abstract
Introduction
Doping controls have become an integral part of professional sport and, with the growing spectrum of drug candidates, approved therapeutics as well as obsolete medicines with purported or evident performance-enhancing properties, the demands and burdens for both athletes and sports drug testing laboratories continue to increase constantly [1]. Consequently, the desire for solutions providing the required analytical information under acceptable circumstances of controls is manifest. Options to complement and/or substitute conventional doping control strategies and corresponding test matrices (i.e. blood and urine) have therefore frequently been assessed [2], [3], [4], especially taking into account accepted strategies employed in diagnostics, forensics, and toxicology. Among the alternative biological matrices, particularly hair, oral fluid (OF), dried blood spots (DBS), and exhaled breath (EB) have received substantial attention, all of which offering advantages but also exhibiting limitations when compared to currently employed routine doping control samples consisting primarily of urine and blood (including whole blood and serum). The use of alternative specimens in sports drug testing is generally warranted as long as the results are not “used to counter Adverse Analytical Findings or Atypical Findings from urine” as specified in paragraph 5.2.4.4 of the World Anti-Doping Code [5]. Therefore, recent accomplishments in developing new analytical approaches using the aforementioned alternative matrices are summarized, and expectations, demonstrated added value, as well as caveats and undisputed downsides are discussed in this mini-review.
Section snippets
Aspects relevant for athletes, doping control laboratories, and anti-doping organizations
The classification of criteria characterizing a specific doping control sampling method is inevitably influenced and colored by the role of the participating party. Even under the assumption that the athlete as well as the anti-doping organization and laboratory pursue the same goal with comprehensive doping controls, i.e. providing evidence that the athlete’s performance was achieved without illicit means, aspects considered relevant for these parties might differ substantially given the
Established test methods in doping controls employing alternative matrices
Numerous doping control analytical methods have been established for urine and blood (including serum and plasma) in the past, which allow for covering the majority of compounds and methods of doping banned in sports according to WADA’s Prohibited List [25]. As summarized in Table 2, also complementary assays employing alternative matrices have been established for various classes of prohibited substances, and the potential as well as limitations of these specimens concerning specific
Conclusion
Doping controls are an integral part of today’s elite sport. They have been the subject of continuous improvements to enable utmost comprehensiveness, aiming at counteracting the fraudulent creativity of some individuals trying to gain the competitive edge via illicit means. Best-possible test methods are required especially in professional sport and, while the combined use of blood and urine does provide the majority of the desired information, alternative matrices have shown to possess the
Acknowledgments
The project was conducted with support of Antidoping Switzerland (Berne, Switzerland) and the Federal Ministry of the Interior of the Federal Republic of Germany (Bonn, Germany).
References (142)
- et al.
Method validation and application of a liquid chromatography–tandem mass spectrometry method for drugs of abuse testing in exhaled breath
J. Chromatogr. B Anal. Technol. Biomed. Life Sci.
(2015) - et al.
Parallel analysis of stimulants in saliva and urine by gas chromatography/mass spectrometry: perspectives for in competition anti-doping analysis
Anal. Chim. Acta
(2008) - et al.
Analytical approaches for the detection of emerging therapeutics and non-approved drugs in human doping controls
J. Pharm. Biomed. Anal.
(2014) - et al.
Mass spectrometric studies on the in vivo metabolism and excretion of SIRT1 activating drugs in rat urine dried blood spots, and plasma samples for doping control purposes
J. Pharm. Biomed. Anal.
(2014) - et al.
A new sulphate metabolite as a long-term marker of metandienone misuse
Steroids
(2013) - et al.
New potential markers for the detection of boldenone misuse
J. Steroid Biochem. Mol. Biol.
(2012) - et al.
Alternative long-term markers for the detection of methyltestosterone misuse
Steroids
(2013) - et al.
Detection and mass spectrometric characterization of novel long-term dehydrochloromethyltestosterone metabolites in human urine
J. Steroid Biochem. Mol. Biol.
(2012) - et al.
The characterisation of selected drugs with amine-containing side chains using electrospray ionisation and ion trap mass spectrometry and their determination by HPLC-ESI–MS
J. Pharm. Biomed. Anal.
(2004) - et al.
Investigations into the feasibility of routine ultra high performance liquid chromatography–tandem mass spectrometry analysis of equine hair samples for detecting the misuse of anabolic steroids, anabolic steroid esters and related compounds
Anal. Chim. Acta
(2013)
Detection of testosterone and epitestosterone in human hair using liquid chromatography–tandem mass spectrometry
J. Pharm. Biomed. Anal.
Ultra high performance liquid chromatography–tandem mass spectrometry determination and profiling of prohibited steroids in human biological matrices. A review
J. Chromatogr. B
Quantification of clenbuterol at trace level in human urine by ultra-high pressure liquid chromatography–tandem mass spectrometry
J. Chromatogr. A
Use of dried blood spots in doping control analysis of anabolic steroid esters
J. Pharm. Biomed. Anal.
Proteome analysis of human hair shaft: from protein identification to posttranslational modification
Mol. Cell. Proteom.
Development and validation of an UPLC-MS/MS method for the quantification of tamoxifen and its main metabolites in human scalp hair
J. Pharm. Biomed. Anal.
Ultra-high performance liquid chromatography–tandem mass spectrometric method for the determination of tamoxifen N-desmethyltamoxifen, 4-hydroxytamoxifen and endoxifen in dried blood spots–development, validation and clinical application during breast cancer adjuvant therapy
Talanta
Solid-phase extraction of acetazolamide from biological fluids and subsequent analysis by high-performance liquid chromatography
J. Chromatogr.
Development and validation of a dried blood spot-LC-APCI-MS assay for estimation of canrenone in paediatric samples
J. Chromatogr. B Anal. Technol. Biomed. Life Sci.
Bioavailability of hydrochlorothiazide from tablets and suspensions
J. Pharm. Sci.
Determination of methylphenidate and ritalinic acid in blood, plasma and oral fluid from adolescents and adults using protein precipitation and liquid chromatography–tandem mass spectrometry—a method applied on clinical and forensic investigations
J. Pharm. Biomed. Anal.
Prescription and illicit psychoactive drugs in oral fluid–LC–MS/MS method development and analysis of samples from Brazilian drivers
Forensic Sci. Int.
Development and validation of a single LC–MS/MS assay following SPE for simultaneous hair analysis of amphetamines opiates, cocaine and metabolites
Forensic Sci. Int.
Analytical strategy for detecting doping agents in hair
Forensic Sci. Int.
On-line liquid chromatography/tandem mass spectrometry simultaneous determination of opiates cocainics and amphetamines in dried blood spots
J. Chromatogr. B Anal. Technol. Biomed. Life Sci.
Simplifying sample pretreatment: application of dried blood spot (DBS) method to blood samples including postmortem, for UHPLC–MS/MS analysis of drugs of abuse
Forensic Sci. Int.
Annual banned-substance review: analytical approaches in human sports drug testing
Drug Test. Anal.
Potential and limitations of alternative specimens in doping control
Bioanalysis
Microsample analyses via DBS: challenges and opportunities
Bioanalysis
The potential role of oral fluid in antidoping testing
Clin. Chem.
Blood analysis and doping control: legal, social, and organizational issues
The fight against doping: back on track with blood
Drug Test. Anal.
Detecting peptidic drugs, drug candidates and analogs in sports doping: current status and future directions
Expert Rev. Proteom.
Dried blood spots (DBS) for doping control analysis
Drug Test. Anal.
Sensitive determination of prohibited drugs in dried blood spots (DBS) for doping controls by means of a benchtop quadrupole/orbitrap mass spectrometer
Anal. Bioanal. Chem.
Detection of Delta9-tetrahydrocannabinol in exhaled breath collected from cannabis users
J. Anal. Toxicol.
Cannabinoids in exhaled breath following controlled administration of smoked cannabis
Clin. Chem.
Quantification of cocaine and metabolites in exhaled breath by liquid chromatography–high-resolution mass spectrometry following controlled administration of intravenous cocaine
Anal. Bioanal. Chem.
Significant increase of salivary testosterone levels after single therapeutic transdermal administration of testosterone: suitability as a potential screening parameter in doping control
Drug Test. Anal.
Significant enhancement of 11-hydroxy-THC detection by formation of picolinic acid esters and application of liquid chromatography/multi stage mass spectrometry (LC–MS(3)): application to hair and oral fluid analysis
Drug Test. Anal.
Use of alternative specimens: drugs of abuse in saliva and doping agents in hair
Ther. Drug Monit.
Hair analysis for drug detection
Ther. Drug Monit.
Hair as an alternative matrix in bioanalysis
Bioanalysis
Hair-based rapid analyses for multiple drugs in forensics and doping: application of dynamic multiple reaction monitoring with LC–MS/MS
Chem. Cent. J.
Statistical significance of hair analysis of clenbuterol to discriminate therapeutic use from contamination
Drug Test. Anal.
Fragmentation studies of SIRT1-activating drugs and their detection in human plasma for doping control purposes
Rapid Commun. Mass Spectrom.
Cited by (95)
Liquid vs dried blood matrices: Application to longitudinal monitoring of androstenedione, testosterone, and IGF-1 by LC–MS-based techniques
2024, Journal of Pharmaceutical and Biomedical AnalysisEvidence of ostarine excretion in oral fluid after a single controlled oral administration
2024, Clinica Chimica ActaA comprehensive review on current analytical approaches used for the control of drug abuse in sports
2023, Microchemical JournalDoping with testosterone and androgenic/anabolic steroids: Impact on health, screening tools and medical care
2023, Annales d'EndocrinologieCapillary blood as a complementary matrix for doping control purposes. Application to the definition of the individual longitudinal profile of IGF-1
2023, Journal of Pharmaceutical and Biomedical Analysis