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EC number: 201-549-0
CAS number: 84-65-1
Table No.6. Summary
Figures with asterisk = values
statistically significant on probability level 0.05 (Mann-Whitney test)
Figures with cross = values ≥ 3
NC – Negative control group
PC – Positive control group
test substance, Anthraquinone, was tested for the assessment of skin
sensitisation potential with the murine local lymph node assay. This
study is a part of the test substance health hazard evaluation.
Local Lymph Node Assay (LLNA) with radionuclides was used. The testing
was conducted according to the EU Method B.42, Skin sensitization: Local
Lymph Node Assay, Council Regulation (EC) No. 440/2008, published in
O.J. L142, 2008.
this study the contact allergenic potential of Anthraquinone was
evaluated after topical application to female BALB/c mice. Five mice per
group were exposed by test and control substances on the dorsum of both
ears once a day during 3 consecutive days. Primary proliferation of
lymphocytes in the lymph node draining the site of application was
evaluated on the base on using of radioactive labelling. The ratio of
the proliferation in treated groups to that in vehicular controls,
termed the Stimulation Index, was determined. Statistical evaluation of
ear weight was performed for elimination of false positive findings with
certain skin irritants.
positive control DNCB (dinitrochlorobenzene): 0.5% (w/v) and
Antraquinone: 30%, 3%, 0.3% (w/v) in DAE 433.
animals exposed to the test substance at the lowest and highest dose
levels showed no pathological skin reactions and no other negative
clinical symptoms of intoxication throughout the experiment. Slight
hyperaemia of skin and swelling of ears was recorded at the highest dose
level. There was no significant difference in body weight increment of
all groups in comparison to the vehicle control.
positive control substance DNCB elicited a reaction pattern with
statistically significant increase in ear weight and Stimulation Index
of cell proliferation 10.5, which was in congruence with his expected
mode of action as a contact allergen.
test substance Anthraquinone showed a tendency to increase ear weight
with concentration-dependence. Ear weight of animals in all dose level
was statistically significantly increased. The result of skin irritation
effect was considered as positive – it means the test substance caused
irritation of skin.
of Stimulation Indexes between treated groups and control group revealed
that the test substance Anthraquinone caused significant increase in
radioisotope incorporation into the DNA of dividing lymphocytes. This
effect was dose dependent.
test substance Anthraquinone had a positive result in LLNA test. Positive
results in cell proliferation and no clinical symptoms of systemic
toxicity revealed that the test substance Anthraquinone could be a
contact allergen in mice but potential irritation effect does not rule
out the possibility that it could be false positive result.
on the positive results of LLNA study, for the test
substance Anthaquinone, it could be recommended to be assigned by symbol
“Xi“ and the risk phrase R43 “may cause sensitisation by skin contact”.
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