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Isolation and Quantitative Determination of New Tuberculostatic 1,2,4-Triazole Derivative in Urine and Plasma Samples

Malgorzata Tatarczak- Michalewska, Jolanta Flieger, Monika Wujec and Marta Swatko- Ossor

A novel series of 1,2,4-triazole derivatives, have been synthesised. All synthesised compounds were screened for their antimycobacterial activity against Mycobacterium tuberculosis and antimicrobial activities against various bacteria and fungi. Among studied derivatives, 4-methyl-1-morpholinmethyl-2,4-dihydro-3H-1,2,4-triazol-3-thione was found as the most active, demonstrating inhibition of mycobacterial growth of Mycobacterium H37Ra at the same concentration level as Streptomycin. The quantitative analysis of this compound in the urine and serum samples was elaborated on. Quantification was performed by reversed-phase high-performance liquid chromatography (RP–HPLC) using C18 column with a mobile phase consisting of methanol and water (5:95) at flow rate of 1 ml/min. The chromatograms were recorded at 245 nm at 21°C. The chromatographic peak identification was based on the comparison the retention times and UV spectra of isolated analyte and synthetized standard. Moreover, different sample pre-treatment methods such as deproteinization and solid-phase extraction (SPE) were compared. The highest analyte recoveries ranging from 78,7% to 90,7% were obtained for sample deproteinized by 6% HClO₄.