Urine Testing to determine use of Physically Enhancing Drugs Jonathan Mertz William Kou Argument • Urine testing is a valid method of determining whether athletes have been using illicit drugs, and whether the urine sample has been tampered with. Overview of the Kidney and Why Urine is a good representation of Substances in the Body • The kidneys are the body’s filtration system. It uses a highly permeable blood vessel system which allows wastes, metabolites, water, toxins, and excess substances to exit the blood vessel. • The Kidneys then reabsorbs the metabolites and water which can be reused and circulated through the blood. • This leaves the toxins, waste and excess substances which the body does not require. • Among these, physical enhancement drugs such as Steroids can be found. Urine Collection • Any athlete participating in a sports event supervised by WADA must be subject to a random urine analysis test. • During collection, there must be a witness present as well as a Doping Control Officer who retains the right to have the athlete remove or adjust any clothing which would impede sample provision • The sample is then sealed into two tamper evident bottles, totaling a minimum of 90 mL of sample Other Collection Requirements • “WADA accredited laboratories are required […] to analyze a minimum of 3000 doping samples provided annually by code compliant testing authorities.” • Athletes are required to complete a medical declaration which “states all medicines, drugs and substances taken over the last week.” • The samples are required to be kept for a minimum of 3 months after the sample was taken. Lab Testing • • • • • • Specific Gravity Test GC/MS Urinary Dipstick Test 1D-Gel Electrophoresis DNA Typing LC-MS/MS Analysis Urinary Dipstick Test • A screening test which provides easy determination of a sample’s specific gravity, among other factors. Gel Electrophoresis Cases of Tampering Attempts • An unremarkable urine sample was run through the screening process (GS-MS/NPD) and found to have none of the hormones which are expected in human urine. The sample was submitted to an ethanol test and determined to be dealcoholized beer. Red Urine Samples • Three urine samples were obtained which were a somewhat uncommon red color. • After screening tests, one of the samples was shown to have blood in it, the others went on to further tests. • After LC-MS, the results showed that Betanin (the dye from beets) was the cause of the red color of the urine. • It was determined that the sample was not tampered with Famous Athletes Caught Using Urine Test • Ben Johnson • Caught with Stanozolol, which is an anabolic steroid, in his urine . • Alberto Contador tested positive for Clenbuterol in his urine • Bryan Berard tested positive for Norandrosterone in a urine test • Floyd Landis was caught with abnormally high levels of testosterone in his urine. Techniques used • • • • • Specific Gravity Test Urinary Dipstick Test 1D-Gel Electrophoresis DNA Typing LC-MS/MS Analysis • Steroids take roughly 30 days to completely leave the body