title: In vitro metabolic stability and metabolite profiling of TJ0711 hydrochloride, a newly developed vasodilatory β-blocker, using a liquid chromatography-tandem mass spectrometry method.
abstract: In this paper, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous analysis of metabolic stability and metabolite profiling of 1-[4-(2-methoxyethyl) phenoxy]-3-[[2-(2-methoxyphenoxy) ethyl]amino]-2-propanol hydrochloride (TJ0711 HCl), a new vasodilatory β-blocker. Multiple reaction monitoring (MRM) was used as a survey scan to quantify the parent compound and to trigger the acquisition of enhanced product ions (EPI) for the identification of formed metabolites. In addition, comparison between MRM-only and MRM-information dependent acquisition-EPI (MRM-IDA-EPI) methods was conducted to determine analytical variables, including linearity, limit of detection (LOD), lower limit of quantification (LLOQ), as well as intra-day and inter-day accuracy and precision. Results demonstrated that MRM-IDA-EPI quantitative analysis was not affected by the addition of EPI scans to obtain qualitative information during the same chromatographic run, compared to MRM-only method. Thereafter, metabolic stability and metabolite identification of TJ0711 HCl were investigated using human liver microsomes (HLM) by the MRM-IDA-EPI method. The in vitro metabolic stability parameters were calculated and t(1/2), microsomal intrinsic clearance (CL(int)), as well as hepatic CL, were 13.0 min, 106.5 μL/min/mg microsomal protein, and 1082.2 mL/min, respectively. The major formed metabolites were also simultaneously monitored and the metabolite profiling data demonstrated that this MRM-IDA-EPI method was capable of targeting a large number of metabolites, in which demethylation and hydroxylation were the principle metabolism pathways during the in vitro incubation with HLM.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Jiangeng
authors.initials: J
authors.lastname: Huang
authors.forename: Luqin
authors.initials: L
authors.lastname: Si
authors.forename: Zhaoze
authors.initials: Z
authors.lastname: Fan
authors.forename: Lei
authors.initials: L
authors.lastname: Hu
authors.forename: Jun
authors.initials: J
authors.lastname: Qiu
authors.forename: Gao
authors.initials: G
authors.lastname: Li
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000032:analysis / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D006900:Hydroxylation; D016014:Linear Models; D008745:Methylation; D008862:Microsomes, Liver / Q000378:metabolism; D050198:Phenoxypropanolamines / Q000032:analysis* / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014665:Vasodilator Agents / Q000032:analysis* / Q000378:metabolism*
chemical_list: C558139:1-(4-(2-methoxyethyl)phenoxy)-3-(((2-2-methoxyphenoxy)ethyl)amino)-2-propanol; D000319:Adrenergic beta-Antagonists; D050198:Phenoxypropanolamines; D014665:Vasodilator Agents