title: Hyphenation of reverse-phase HPLC and ICP-MS for metabolite profiling--application to a novel antituberculosis compound as a case study.
abstract: In this study, a high-performance liquid chromatography (HPLC) inductively coupled plasma (ICP) mass spectrometry (MS) method was developed intended for use in metabolism studies of bromine-containing drugs, administered to test animals (or test persons). As a case study, the method was applied to a new antituberculosis compound, the bromine-containing diarylquinoline R207910. A method has been proposed to overcome the incompatibilities between the high organic solvent content (45%CH3OH and 45% CH3CN) used in the reverse-phase liquid chromatography (LC) separation on one hand and the limitations of the ICP on the other hand. Therefore, several instrument modifications had to be made. For the introduction of the column effluent, a combination of a perfluoroalkoxy LC nebulizer with a PC(3) Peltier-cooled inlet system (operated at 2 degrees C) was used. Additionally, the standard injector tube (internal diameter 2 mm) was replaced by an injector tube with an internal diameter of 1 mm and to avoid carbon build-up on the interface cones and the torch, the nebulizer gas was admixed with 6% v/v of oxygen. After optimization of the method, HPLC-ICP-MS was applied for metabolite profiling of faeces samples after dosing of (14)C-radiolabelled R207910 to dogs and rats. To evaluate the method developed, the HPLC-ICP-MS results were compared with those of HPLC with UV spectrophotometric and (14)C radiochemical detection. As the HPLC-ICP-MS method gave rise to a higher selectivity than HPLC with UV detection and to a better detection limit (5 ng R207910) than the method with radiochemical detection (65 ng R207910), it can be concluded that ICP-MS can be used as a good alternative to the more traditional detection methods, even when a mobile phase with high organic solvent content has to be used in the LC separation.
authors.affiliation: Department of Analytical Chemistry, Ghent University, Proeftuinstraat 86, 9000 Ghent, Belgium.
authors.forename: Lieve I L
authors.initials: LI
authors.lastname: Balcaen
authors.forename: Björn
authors.initials: B
authors.lastname: De Samber
authors.forename: Kenny
authors.initials: K
authors.lastname: De Wolf
authors.forename: Filip
authors.initials: F
authors.lastname: Cuyckens
authors.forename: Frank
authors.initials: F
authors.lastname: Vanhaecke
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000032:analysis*; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D064687:Diarylquinolines; D004285:Dogs; D004355:Drug Stability; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D009930:Organic Chemicals / Q000737:chemistry; D004364:Pharmaceutical Preparations / Q000032:analysis*; D011804:Quinolines / Q000032:analysis; D051381:Rats; D012997:Solvents / Q000737:chemistry; D013696:Temperature
chemical_list: D000995:Antitubercular Agents; D002250:Carbon Radioisotopes; D064687:Diarylquinolines; D009930:Organic Chemicals; D004364:Pharmaceutical Preparations; D011804:Quinolines; D012997:Solvents; C493870:bedaquiline