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.
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).
Eggenreich
K
K
Institute of Pharmaceutical Chemistry and Pharmaceutical Technology, Karl-Franzens-University, Schubertstrasse 1 A-8010, Graz, Austria.
Golouch
S
S
Töscher
B
B
Beck
H
H
Kuehnelt
D
D
Wintersteiger
R
R
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*
D003408:Cresols; D006202:Hair Dyes