Development and validation of an LC–MS/MS method for determination of methanesulfonamide in human urine

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 877; no. 22; pp. 2087 - 2092
• Main Authors: Anacardio, Roberto, Mullins, Frank G.P., Hannam, Sally, Sheikh, Muhammed S., O'Shea, Karen, Aramini, Andrea, D’Anniballe, Gaetano, D’Anteo, Loredana, Ferrari, Mauro P., Allegretti, Marcello
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.07.2009; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Humans; Interleukin-8 - antagonists & inhibitors; LC–MS/MS (liquid chromatography–mass/mass); Medical sciences; Methanesulfonamide (MSA); Pharmacology. Drug treatments; Reparixin; Sulfonamides - metabolism; Sulfonamides - urine; Tandem Mass Spectrometry - methods; Urine; Urine; Methanesulfonamide (MSA); Reparixin; LC–MS/MS (liquid chromatography–mass/mass); Human; Validation; Biological fluid; Sulfonamide; Antiinflammatory agent; HPLC chromatography; Determination; LC-MS/MS (liquid chromatography-mass/mass); Mass spectrometry MS/MS; Development; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2009.05.051

A sensitive and selective liquid chromatographic method coupled with tandem mass spectrometry (LC–MS/MS) was developed and validated for the quantification of methanesulfonamide (MSA) in human urine. MSA is a potential in vivo metabolite of reparixin, a specific inhibitor of the CXCL8 biological activity. In this study, a simple derivatization procedure with a new reagent, N-(4-methanesulfonyl-benzoyl)-imidazole, was set up to enable MSA and the internal standard (I.S.), ethanesulfonamide (ESA), to be analysed by LC–MS/MS. After derivatization, samples were evaporated and reconstituted in 30% acetonitrile, aq. MSA and I.S. derivatives were separated by reversed phased HPLC (high performance liquid chromatography) on a Luna 5μ C18 column and quantitated by MS/MS using electrospray ionization (ESI) and multiple reaction monitoring (MR M) in the negative ion mode. The most intense [M−H]− MRM transition of derivatized MSA at m/z 276.2→197.2 was used for quantitation and the transition at m/z 290.2→211.2 was used to monitor derivatized ESA. The method was linear over the concentration range from 1 to 100μg/ml, with a lower limit of quantitation of 1μg/ml. The intra- and inter-day precisions were less than 5.5% and 10.1%, respectively, and the accuracies were between −4.0% and +11.3%. The method was successfully applied to quantify levels of MSA in human urine after intravenous administration of reparixin to healthy volunteers.