Capillary ultra performance liquid chromatography–tandem mass spectrometry analysis of tienilic acid metabolites in urine following intravenous administration to the rat

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2018-06, Vol.1087-1088, p.142-148
• Main Authors: King, Adam M., Grant, Isobelle, Rainville, Paul D., Isaac, Giorgis, Coen, Muireann, Wilson, Ian D., Plumb, Robert S.
• Format: Article
• Language: English
• Subjects: Bioanalysis; Capillary UPLC; DMPK; Metabolites; Tienilic acid
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2018.04.034

Capillary scale (100 mm × 150 μm id) UPLC/MS/MS, performed using reversed-phase gradient chromatography on sub 2 μm particles, has been successfully employed for the characterization of the metabolites of the drug tienilic acid (TA) excreted via the urine following oral administration to the rat. The capillary LC system provided a significant increase (range ca. 11–33-fold) in sensitivity compared with a conventional 150 mm × 2.1 mm id UPLC system. An investigation of the effect of the injection volume and sample mass loading on the capillary column on the results obtained for both endogenous metabolites and TA was performed. This demonstrated that the injection of up to 2 μL of rat urine onto the system was permitted whilst still providing excellent chromatographic results and robustness. Qualitative analysis of the urine revealed the presence of TA itself and a total of 15 metabolites of the drug, including those resulting from biotransformations such as hydroxylation or conjugation. The capillary chromatography system was shown to be robust, and capable of providing comprehensive drug metabolite profiles from small format urine samples such as those obtained from preclinical studies in rodents. •Reversed-phase gradient capillary scale U(H)PLC/MS/MS on sub 2 µm particles successfully enabled the analysis of rat urine for drug metabolites.•A total of 15 metabolites of the drug tienilic acid were detected including hydroxylated and conjugated compounds.•Capillary LC was shown to be 11-38-fold more sensitive than the conventional system for these metabolites.•144 injections of 2µL of rat urine were analysed in a single run without adverse effects on system performance.