Identyfikacja diuretyków w moczu techniką ultrasprawnej chromatografii cieczowej połączonej z tandemową spektrometrią mas (UPLC/MS/MS)

Diuretics are medicines that intensify the diuresis process. There are several types of diuretics (high ceiling loop diuretics, thiazides, calcium-sparing diuretics, potassium-sparing diuretics, osmotic diuretics). Generally all of them increase excretion of water from a body. According to the Prohibited List published by the World Anti-Doping Agency diuretics make a group of substances which is prohibited in sport at all times (in- and out-of-competition). This review describes shortly an ultra performance liquid chromatography technique, chemical properties of diuretics named in the WADA Prohibited List as well as a developed analytical method with the use of UPLC/MS/MS technique for determination of diuretics and other doping agents (stimulants, masking agents) in urine samples in comparison with a GC/MS method used earlier. Detailed extraction procedure and comparison of application of several organic solvents during extraction process were given. Parameters of MS/MS method have also been given. The described method was validated in accordance with EURACHEM "The Fitness for Purpose of Analytical Methods: A Laboratory Guide to Method Validation and Related Topics". Limits of detection and recovery level for all compounds in the method were evaluated. The developed method allows to detect most of diuretics at the levels of 1/5 MRPL (Minimum Required Performance Limits) and fulfills all requirements of the World Anti-Doping Agency.