Isolation of the mutagenic and DNA adduct-inducing components from a commercial preparation of HC blue 1 using Salmonella (TA98) bioassay-directed HPLC fractionation.
In the present study we report the separation of the mutagenic impurities from the nitrophenylenediamine hair dye HC Blue 1. This was accomplished by bioassay-directed HPLC fractionation, using Salmonella strain TA98 and reverse phase HPLC analysis. The mutagenic fraction eluted between 80 and 90% methanol, whereas the HPLC fraction containing the parent compound HC Blue 1 eluted with 30% methanol and was non-mutagenic. 100% of the mutagenic activity applied to the column was recovered in fractions that did not possess the blue color of HC Blue 1. Also, HPLC-purified HC Blue 1 did not form DNA adducts (32P-postlabeling) in Salmonella strain TA98. On the other hand, commercial HC Blue 1 and the mutagenic fraction derived from commercial HC Blue 1 (HPLC-isolated) gave similar DNA-adduct profiles that consisted of 7 adducts. DNA adduction was examined concomitantly with mutagenicity and toxicity studies on the HC Blue 1 samples in TA98. The data indicated that, in Salmonella, both the mutagenicity and DNA adduction of commercial HC Blue 1 are due to impurities and not the parent compound.
Abu-Shakra
A
A
National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.
Johnson
L
L
Earley
K
K
Jameson
C W
CW
Kari
F W
FW
Gupta
R
R
Langenbach
R
R
D001345:Autoradiography; D002851:Chromatography, High Pressure Liquid; D004249:DNA Damage; D006202:Hair Dyes / Q000633:toxicity*; D009152:Mutagenicity Tests; D009153:Mutagens; D010655:Phenylenediamines / Q000633:toxicity*; D012486:Salmonella typhimurium / Q000187:drug effects; D013056:Spectrophotometry, Ultraviolet
D006202:Hair Dyes; D009153:Mutagens; D010655:Phenylenediamines; C007906:N-methylamino-2-nitro-4-N',N'-bis(2-hydroxyethyl)aminobenzene