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2021, Number 1

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Rev Cubana Farm 2021; 54 (1)

Importance of samples preparation in the study of clobenzorex’s metabolism

McPherson MA, Martínez BD, Torres CM, Fiallo FT, Correa VT, Montes de Oca PR

Language: Spanish
References: 30
Page: 1-21
PDF size: 1558.92 Kb.


ABSTRACT

Introduction: The selection of the trial to study the metabolism of a substance depends on its metabolic reactions, which can positively influence the selectivity of the method and the collection of analytes, among others.
Objective: Define the influence of sample preparation in the study of clobenzorex metabolism in the conditions of the Anti-Doping Laboratory in Havana.
Methods: Urine samples were collected before and after the administration of a dose of clobenzorex to a supposedly healthy volunteer. Three trials (A, B and C) designed for the detection of Phase I and Phase II metabolites were applied. The extraction method was liquid-liquid and the difference between the methods was the derivatives formation and enzymatic hydrolysis. The instrumental technique used was gas chromatography coupled to mass spectrometry, and the data collection mode was SCAN from 50 to 600 Da.
Results: The detection of the most volatile clobenzorex’s metabolites was affected by the manipulation of the sample, while the hydroxylated ones (phase I) required hydrolysis to be detected. With the application of method A, 3 compounds were identified, 5 with the application of method B and with the application of method C, 8 metabolites were identified. At 24 hours after administration of the drug, 6 of these compounds could be detected by method C.
Conclusions: The application of appropriate sample preparation methods, not only to detect doping in athletes, but also in the study of the metabolism of drugs eliminated in urine, defines the quantity and quality of the information obtained. Under the conditions of the Anti-Doping Laboratory of Havana, better detection of clobenzorex and its metabolites requires a step of enzymatic hydrolysis and one of derivative formation, in order to obtain analytes with good chromatographic conditions when analyzed by the CG-EM technique.


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