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Quantification of 1‐(propan‐2‐ylamino)‐4‐propoxy‐9 H ‐thioxanthen‐9‐one (TX5), a newly synthetized P‐glycoprotein inducer/activator, in biological samples: method development and validation
Author(s) -
Ferreira Ana Filipa,
Ponte Filipa,
Silva Renata,
RochaPereira Carolina,
Sousa Emília,
Pinto Madalena,
Bastos Maria de Lourdes,
Remião Fernando
Publication year - 2017
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3802
Subject(s) - chemistry , activator (genetics) , inducer , chromatography , glycoprotein , biochemistry , gene
A simple, rapid and economical method was developed and validated for the analysis and quantification of 1‐(propan‐2‐ylamino)‐4‐propoxy‐9 H ‐thioxanthen‐9‐one (TX5), a P‐glycoprotein inducer/activator, in biological samples, using reverse‐phase high‐performance liquid chromatography (HPLC). A C 18 column and a mobile phase composed of methanol–water (90/10, v /v) with 1% (v/v) triethylamine, at a flow rate of 1 mL/min, were used for chromatographic separation. TX5 standards (0.5–150 μ m ) were prepared in human serum. Methanol was used for TX5 extraction and serum protein precipitation. After filtration, samples were injected into the HPLC apparatus and TX5 was quantified by a conventional UV detector at 255 nm. The TX5 retention time was 13 min in this isocratic system. The method was validated according to ICH guidelines for specificity/selectivity, linearity, accuracy, precision, limits of detection and quantification (LOD and LOQ) and recovery. The method was proved to be selective, as there were no interferences of endogenous compounds with the same retention time of TX5. Also, the developed method was linear ( r 2 ≥ 0.99) for TX5 concentrations between 0.5 and 150 μ m and the LOD and LOQ were 0.08 and 0.23 μ m , respectively. The results indicated that the reported method could meet the requirements for TX5 analysis in the trace amounts expected to be present in biological samples.