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EC number: 272-729-4
CAS number: 68910-05-4
A complex residuum from the fractionation of the reaction products of 2-aminoethanol with ammonia to remove piperazine. It may contain such compounds as 2-[(2-aminoethyl)amino]ethanol, (aminoethyl)piperazine, (hydroxyethyl)piperazine.
* Precipitation in culture medium at the end of exposure period
** Mutant frequency MFcorr.: mutant colonies per 106
cells corrected with the CE2 value
*** Cloning efficiency related to the respective vehicle control
The test substance Amix 1000 was assessed for its potential to induce
gene mutations at the hypoxanthine-guanine phosphoribosyl transferase
(HPRT) locus in Chinese hamster ovary (CHO) cells in vitro. Two
independent experiments were carried out, both with and without the
addition of liver S9 mix from phenobarbital- and β-naphthoflavone
induced rats (exogenous metabolic activation).
According to an initial range-finding cytotoxicity test for the
determination of the experimental doses, the following concentrations
were tested and evaluated.
without S9 mix (4-hour exposure period)
0; 625.0; 1 250.0; 2 500.0; 5 000.0 μg/mL
with S9 mix (4-hour exposure period)
0; 937.5; 1 875.0; 3 750.0; 5 000.0 μg/mL
Following attachment of the cells for 20-24 hours, cells were treated
with the test substance for 4 hours in the absence and the presence of
metabolic activation. Subsequently, cells were cultured for 6-8 days and
then selected in 6-thioguanine-containing medium for another week.
Finally, the colonies of each test group were fixed with methanol,
stained with Giemsa and counted.
The vehicle controls gave mutant frequencies within the range expected
for the CHO cell line. Both positive control substances, EMS and DMBA,
led to the expected increase in the frequencies of forward mutations. In
this study in the absence and the presence of metabolic activation no
cytotoxicity was observed up to the highest required concentration
evaluated for gene mutations.
Based on the results of the present study, the test substance did not
cause any relevant increase in the mutant frequencies either without S9
mix or after the addition of a metabolizing system in two experiments
performed independently of each other.
Thus, under the experimental conditions of this study, the test
substance Amix 1000 is not mutagenic in the HPRT locus assay under in
vitro conditions in CHO cells in the absence and the presence of
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