LC-MS/MS identification of trimetazidine metabolites in human urine

Authors: Kamila Chomaničová 1    Élida Alechaga Silva 2    Alejandro Terrero González 2    Štefan Husár 1    Rosa Ventura Alemany 2   
1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University in Bratislava, SVK    2 Catalonian Anti‐Doping Laboratory, Doping Control Research Group, IMIM, Barcelona, ESP   
Year: 2021
Section: Analytics in biological sciences
Abstract No.: 2100
ISBN: ISBN 978-80-972360-7-6

Application of liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify trimetazidine (TMZ) metabolites in human urine has allowed to study known and unknown metabolites [1]. TMZ is predominantly eliminated in the urine with elimination half‐life of 6.5 hours, without cumulative dose effect [2]. Due to its physicochemical properties TMZ and its metabolites can be retained and separated in a C18 stationary phase by reversed phase chromatography [3]. In the presented study, urine samples of one healthy volunteer have been collected after a single dose of TMZ at the baseline and at the following intervals:  0–4h, 4–8h, 8–12h, 12–24h after administration. Chromatographic separation was carried out using UHPLC C18 column combined with a mobile phase gradient starting at 100% aqueous conditions. Accurate mass of possible protonated/ deprotonated ions of TMZ metabolites from the created database was used to look for possible candidates in the full-scan data of the post-administration samples that were not present in the basal samples using TraceFinder software. Full-scan spectra and product ion scan spectra at low and high collision energies (CE 15 eV, 45 eV) of detected possible metabolites were acquired and as a major metabolite was observed intact TMZ with maximum concentration (11,4 μg/ml) in time interval 4-8 h after the administration. Moreover, five minor metabolites have been observed, namely Trimetazidine-N-oxide (M1), N-formyl trimetazidine (M2), Desmethyl-trimetazidine O-sulphate (M3), Desmethyl-trimetazidine O-glucuronide (M4) and Desmethyl-trimetazidine-N-oxide-O-glucuronide (M5), which was the only one up to now unreported . In addition, renal excretion profiles of detected TMZ metabolites have been proposed.

This work was supported by the Grant for PhD students and young researchers at Comenius University no. UK /94/2021.
[1] Sigmund, G., et al. Doping control analysis of trimetazidine and characterization of major metabolites using mass spectrometric approaches. Drug Test Anal. 2014; 6(11-12): 1197-205. doi: 10.1002/dta.1680.
[2] Ciapponi, A., Pizarro, R., Harrison, J. Trimetazidine for stable angina. Cochrane Database of Systematic Reviews. 2017; (3): doi:10.1002/14651858.cd003614.pub3.
[3] Zhou, G., et al. Development and validation of a simple and sensitive liquid chromatography-tandem mass spectrometry method for quantifying trimetazidine in human plasma. Clinical and Experimental Pharmacology and Physiology. 2010; 37(4): 501–505. doi:10.1111/j.1440-1681.2009.05326.x