The UCLA Laboratory is accredited by the World Anti-Doping Agency (WADA), an international independent organization created in 1999 having the self-described mission of promoting, coordinating and monitoring the fight against doping in sport in all its forms. In part, maintaining accreditation with the WADA requires compliance with the International Standard for Laboratories (ISL) maintained by the WADA to ensure production of valid test results and evidentiary data, and to achieve harmonized results and reporting from all accredited laboratories. Among the requirements, as laid out in the ISL, are minimum required performance limits (MRPL) for target compounds and successful completion of proficiency testing each year.
Since 1999 the UCLA Olympic Analytical Laboratory has also been accredited to ISO/IEC 17025 – Certificate 1420.01 (and its precursor ISO/IEC Guide 25) for chemical testing by the American Association for Laboratory Accreditation (A2LA). As of January 1, 2004 WADA has required that all laboratories that are accredited by WADA hold accreditation against ISO/IEC 17025. In order to maintain its A2LA accreditation the laboratory must undergo an annual audit of procedures, test methods and records and in addition must undergo a biennial onsite accreditation assessment.
The UCLA Olympic Laboratory has been a leader in research activities directed towards enhancing methods to detect doping in sports. Research conducted by the UCLA Laboratory and others in the mid 1990's resulted in the development of a gas chromatography-combustion-isotope ratio mass spectrometry (IRMS) method to detect exogenous testosterone administration (1). Adaptations of this technique are currently being utilized to detect a variety of steroids in urine specimens (2-4).
In 1996 the laboratory along with other IOC-accredited laboratories began utilizing high-resolution mass spectrometry (HRMS) to increase the sensitivity and retrospectively of screening and confirmatory tests for steroids including methandienone and stanozolol.
In 2002 the laboratory identified norbolethone, a steroid that had never been marketed, in an athlete's urine (5). In 2004 the laboratory published its findings regarding the isolation of tetrahydrogestrinone (THG), a steroid previously unknown in the literature, from the residue in a syringe anonymously turned over to the United States anti-doping agency (6). In 2005 the laboratory detected methylandrostenol (MADOL), another steroid previously unknown in the literature, in an athlete's urine sample (7).
The laboratory's experience with THG led to the transition to liquid chromatography tandem mass spectrometry (LCMSMS) based detection methods for many steroids, an approach that was found in many respects to be superior to traditional gas chromatography-mass spectrometry (GCMS)-based methods. The methodology for the detection of THG in urine was subsequently distributed to all WADA accredited laboratories.
Anabolic steroids are male sex hormones that promote protein storage and tissue growth. Athletes often abuse them to increase muscle size and strength. Some steroids occur naturally in the body and others are synthetic derivatives of testosterone.
Anti-estrogenic agents block the conversion of testosterone to estrogens and prevent the body from responding to these female hormones. These agents can reduce the unwanted side effects from anabolic steroids (such as infertility and breast development) and make more testosterone available for muscle development.
Beta-2-agonists, such as salbutamol, often used to treat asthma and breathing aliments. One of these agents, clenbuterol, is taken to promote muscle growth and is not legally available for human use in the United States.
Beta-blockers, such as propranolol, lower blood pressure. Athletes abuse them to pursue a competitive edge in sports such as archery and shooting. By slowing the heart rate these compounds can improve hand-eye coordination.
Diuretics are often abused to mask a banned substance, because they speed its elimination from the body via increased urination and water loss.
Erythropoietin (EPO) is a hormone normally produced by the body that stimulates red blood cell production, increasing the delivery of oxygen to muscle tissue.
Glucocorticosteriods, such as prednisone and cortisone are chronically abused to reduce inflammation.
Human chorionic gonadotropin (hCG) is a hormone that enhances the production of male hormones for developing muscle mass.
Stimulants, such as amphetamines, heighten the athlete's ability to train at high levels, suppress the appetite for weight loss, and can increase mental awareness and physical responsiveness.
Street drugs, including opiates, cocaine and marijuana.
The laboratory's testing facility is equipped with a wide range of instruments in order to utilize specific testing methods that are most suitable for drug detection and includes manual and automated immunoassay, gas chromatography-mass spectrometry (GCMS), high-resolution mass spectrometry (HRMS), high performance liquid chromatography (HPLC), GC-combustion-isotope ratio mass spectrometry (IRMS), and liquid chromatography-tandem mass spectrometry (LCMSMS).
With specific regard to GCMS, the technique most commonly employed by WADA accredited laboratories for the detection of anabolic steroids, the facility is currently equipped with 15 instruments comprised of Agilent 6890 gas chromatographs coupled to 5973 and 5975 series single-quadruple mass spectrometers which gives the facility unsurpassed capacity among WADA accredited laboratories. The addition of a Varian 300-MS system has added the capability to carry out GCMSMS analysis.
The facility is equipped with a range of LCMSMS systems including API-3000 triple-quadruple, 4000 Q Trap and QTRAP analyzers, and two IRMS systems for confirmatory analysis of metabolites of endogenous steroids such as testosterone and androstenedione.
Sample preparation for steroid analysis is handled by 15 modular automated solid phase extraction workstations.
The UCLA laboratory conducts all laboratory work according to its extensive and detailed Standard Operating Procedures. All procedures are conducted in accordance with the relevant requirements of the International Olympic Committee, WADA, and ISO/IEC Guide 17025 accreditation through A2LA.
The laboratory maintains chain of custody documentation for samples from the time they are delivered by courier to the facility, to the time they are discarded as directed by the client.
The typical analysis is initiated by aliquoting a portion of the A-sample for a steroid screen by GCMS and/or LCMSMS. Samples producing negative results are reported as negative without additional testing. Should a screen indicate the presence of a proscribed substance (adverse analytical finding), an A-confirmation is performed. Confirmed adverse analytical findings are reported to the client and an analysis of the corresponding B-sample is scheduled.
All adverse analytical findings are accompanied by a documentation package detailing the findings as well as providing relevant supporting information including sample chain of custody records. All reports are considered confidential.