Abstract
A fast and sensitive method for determination of 8 diuretics (acetazolamide, bendroflumethiazide, bumetanide, chlorthalidone, furosemide, hydrochlorothiazide, metolazone, triamterene) and masking agent (probenecid) in human urine using gas-chromatography with mass spectrometric detection is described. The extraction of the substances as function of the nature of organic solvent, mixing time and pH of aqueous phase was studied. The tandem mass spectrometry was used to increase selectivity of diuretics determination due to elimination of background interferences. Fragmentation reactions were studied for each compound and their collision energies were optimized to obtain the best selectivity. The results of method’s validation demonstrate its suitability in routine analysis for confirmation purposes.
Highlights
It is known [1] that athletes misuse diuretics for several reasons: to reduce body weight in order to be qualified for a lower weight category, to decrease fluid retention in body caused by the application of steroids and to reduce the concentration of other prohibited substances in urine to avoid positive doping results
In this work the recovery degree as the function of the nature of organic solvent, mixing time and pH of aqueous phase were investigated to optimize the conditions of diuretics determination
The extraction degree of all substances is satisfactory, though the recovery of metolazone and trichlormethiazide under these conditions is 53.7% and 3.1%, respectively, but the signal intensity is sufficient enough to detect these compounds at the minimum required performance limit (MRPL) level
Summary
It is known [1] that athletes misuse diuretics for several reasons: to reduce body weight in order to be qualified for a lower weight category, to decrease fluid retention in body caused by the application of steroids and to reduce the concentration of other prohibited substances in urine to avoid positive doping results. WADA established the requirement for anti-doping laboratories to control the presence of these compounds in urine at the level less or equal to minimum required performance limit (MRPL), which is 0.25 μg·ml–1 [2]. Most of the banned substances should not be present at any concentration in the specimens (mainly urine) collected for the analysis. They are often detected only on a qualitative basis.
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