title: Development and validation of an LC-MS/MS method for determination of methanesulfonamide in human urine.
abstract: 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 5micro 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 100 microg/ml, with a lower limit of quantitation of 1 microg/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.
authors.affiliation: Dompé pha.r.ma s.p.a., Research Department Via Campo di Pile, L'Aquila, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Anacardio
authors.forename: Frank G P
authors.initials: FG
authors.lastname: Mullins
authors.forename: Sally
authors.initials: S
authors.lastname: Hannam
authors.forename: Muhammed S
authors.initials: MS
authors.lastname: Sheikh
authors.forename: Karen
authors.initials: K
authors.lastname: O'Shea
authors.forename: Andrea
authors.initials: A
authors.lastname: Aramini
authors.forename: Gaetano
authors.initials: G
authors.lastname: D'Anniballe
authors.forename: Loredana
authors.initials: L
authors.lastname: D'Anteo
authors.forename: Mauro P
authors.initials: MP
authors.lastname: Ferrari
authors.forename: Marcello
authors.initials: M
authors.lastname: Allegretti
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D016209:Interleukin-8 / Q000037:antagonists & inhibitors; D013449:Sulfonamides / Q000378:metabolism / Q000652:urine*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D016209:Interleukin-8; D013449:Sulfonamides; C036871:methanesulfonamide; C490707:reparixin