Registration Dossier

Administrative data

Endpoint:
cytotoxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Remarks:
Screening-test.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Principles of method if other than guideline:
With the XTT-test (colourimetric assessment) the cell proliferation and the number of surviving cells are measured after administration of the test
substance.

The assay is based on the cleavage of the yellow tetrazolium salt XTT (sodium 3-[1-(phenylaminocarbonyl)- 3,4-tetrazolium]-bis (4-methoxy-6-
nitro) benzene sulfonicacid hydrate) to form an orange formazan dye by metabolic active cells. Therefore, this conversion only occurs in viable
cells. The formazan dye formed is soluble in aqueous solutions and is directly quantified using a scanning multiwell spectrophotometer (ELISA
reader). This ensures a high degree of accurracy. Cells, grown in a 96 well tissue culture plate, are incubated with the yellow XTT solution (final
concentration 0.3 mg/ml) for 4–24 h).After this incubation period, orange formazan solutionis formed, which is spectrophotometrically quantified using an ELISA plate reader. An increase in number of living cells results in an increase in the overall activity of mitochondrial dehydrogenases in
the sample. This increase directly correlates to the amount of orange formazan formed, as monitored by the absorbance.
GLP compliance:
no
Type of method:
in vitro

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Results and discussion

Applicant's summary and conclusion

Conclusions:
First slight cytotoxic signs were seen at a concentration of 3.000 mM. 10.000 mM DMAPMA were cytotoxic to the hepatocytes under the present test 
conditions.
Executive summary:

The effect of Dimethylaminopropyl methacrylamide on rat liver hepatocytes was tested. The DMAPMA concentrations used
were 0.003, 0.01, 0.030, 0.100, 0.300, 1.000, 3.000 and 10.000 mM at an incubation time of 24 hours at 37 °C. 
The XTT-test is a colorimetric method which determines the cell  proliferation and the number of surviving cells after
incubation with the test substance. First slight cytotoxic signs were seen at a concentration of 3.000 mM. 10.000 mM
DMAPMA were cytotoxic to the hepatocytes under the present test conditions.