Sweat as an alternative sample in doping control

Author(s)

Dayanne Cristiane Mozaner Bordin

• Document type: Doctoral Thesis
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC)
• Defense date: 2018-02-20

Examining board members

Bruno Spinosa de Martinis; David Paul Bishop; Lucas Blanes

• Advisor: Bruno Spinosa de Martinis; Maurício Yonamine

Abstract

Sport has become a major industry of valuable awards and major investments. In an attempt to overcome opponents, many athletes resort to illegal means to improve their performance. The World Antidoping Agency (WADA) prohibits drug use in sport. In addition to the ethical aspects, drug use can cause serious damage to an athletes\' health by directly influencing physiological capacity and removing physical and psychological barriers, and their health damage may be irreversible. Urine is the standard matrix used in doping control; however, these analyses can also be performed on alternative biological samples; such as sweat. Toxicological analysis in sweat samples present some advantages over urine, such as less chance of sample tampering, greater detection window, non-invasive collection, besides the possibility of finding mainly the parent drugs. The use of sweat as an alternative sample also allows obtaining a history of exposure to drug abuse. The identification of doping cases has been an important area of research and development in recent years due to the continued increase in the number of prohibited substances. For the detection of prohibited substances, sophisticated analytical instruments such as liquid (LC), gas (GC) chromatography coupled to mass spectrometry and capillary electrophoresis (CE) are generally used. This thesis focused on the development of methods for evaluation of sweat as an alternative biological matrix for doping control, using different methods for sample preparation and detection to overcome its limitations. In addition, an outline of the profile of drugs used for sports practice and the acute effects caused in the organism of abusers were investigated. A survey was applied to the volunteers to evaluate the adverse effects of androgenic anabolic steroids (AAS), dietary supplements (DS) and multiple drug use. In parallel, it was measured the volunteers hematological and biochemical parameters. Through toxicological urinalysis, the self-report and the parameters measurement within the study revealed that the use of doping agents and drugs are subjected to different factors, which are normally guided by the type of physical activity or aesthetic appearance. This abuse can increase the chances of health problems causing synergistic side effects, increasing the risk to develop diseases. We developed of a method to determine the presence of 13 amphetamines and cocaine related substances and their metabolites in sweat and urine using Disposable Pipette Extraction (DPX) Tips and GC-MS. The validated method was used to analyze 40 urine and sweat samples whose athletes self-reported the use of drugs and/or stimulants. It was verified that all consumed drugs and metabolites detected in urine were also present in sweat samples indicating that sweat is a viable matrix to perform doping tests. We also developed a screening alternative method for stimulants detection in sweat samples using a microchip capillary electrophoresis instrument (Bionalyzer). Although functioning, the method was not sensitive enough to detect the low concentrations of drugs and metabolites present in sweat samples. A one step fully automatized derivatisation and headspace (HS) SPME extraction method followed by GC-MS was developed for the analysis for amphetamine-type drugs and cocaine. The HS-SPME/GC-MS method was used to detect concentrations between 0.1 to 1 ng/mL of the target analytes without any additional sample preparation, showing to present a potential application for routine analysis of drug traces in biological samples, such as urine and sweat (AU)

• FAPESP's process: 13/25601-7 - To assess the sweat as alternative sample for doping control in sport
• Grantee: Dayanne Cristiane Mozaner Bordin
• Support Opportunities: Scholarships in Brazil - Doctorate