title: Determination of 4-amino-m-cresol and 5-amino-o-cresol and metabolites in human keratinocytes (HaCaT) by high-performance liquid chromatography with DAD and MS detection.
abstract: A multi-step gradient HPLC system combined with DAD and MS detection has been developed for the determination of the oxidation hair dyes 4-amino-m-cresol (4-AC) and 5-amino-o-cresol (5-AC) and their metabolites in the alternative testing system human keratinocytes (HaCaT) cell culture. The culture medium induced by 3-methylcholanthrene (3-MC) was fortified with 4-AC or 5-AC and incubated for 24 h at 37 degrees C in order to produce metabolites. After several pre-cleaning steps, further cleaning was done by solid-phase extraction using C18 phenyl cartridges. Optimizing chromatographic conditions, a hybrid-based RP8 column was most suitable for the separation of the metabolites formed in HaCaT. Only one conjugation product, the N-acetylated derivative, could be identified for both 4-AC and 5-AC by LC/DAD/MS. The ionisation technique used for MS analysis was Atmospheric Pressure Ionization (API).
authors.affiliation: Institute of Pharmaceutical Chemistry and Pharmaceutical Technology, Karl-Franzens-University, Schubertstrasse 1 A-8010, Graz, Austria.
authors.forename: K
authors.initials: K
authors.lastname: Eggenreich
authors.forename: S
authors.initials: S
authors.lastname: Golouch
authors.forename: B
authors.initials: B
authors.lastname: Töscher
authors.forename: H
authors.initials: H
authors.lastname: Beck
authors.forename: D
authors.initials: D
authors.lastname: Kuehnelt
authors.forename: R
authors.initials: R
authors.lastname: Wintersteiger
mesh_terms: D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003408:Cresols / Q000032:analysis* / Q000378:metabolism*; D006202:Hair Dyes / Q000032:analysis* / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D003408:Cresols; D006202:Hair Dyes