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EC number: 271-974-4
CAS number: 68647-86-9
Test item concentration % (w/w)
number of lymph nodes
DPM per lymph nodeb)
(1 ml 5% trichloroacetic acid) in duplicate
1 = Control
a) = The
mean value was taken from the figures BG I and BG II
b) = Since
the lymph nodes of the animals of a dose group were pooled, DPM/node was
determined by dividing the measured value by the number of lymph nodes
estimated concentration of the test item required to produce a 3-fold
increase in draining lymph node cell proliferative activity (termed the
EC3 value) can be calculated according to the equation [EC3=(a-c)
[(3-d)/(b-d)] + c]. Here, (a, b) and (c, d) are respectively the
co-ordinates of the two pair of data lying immediately above and below
the S.I. value of 3 on the local lymph node assay dose response plot.
value could not be calculated, since all S.I.´s are below 3.
occurred during the study period.
of local toxicity at the ears of the animals and no systemic findings
were observed during the study period.
weight of the animals, recorded prior to the first application and prior
to treatment with 3HTdR, was within the range commonly recorded for
animals of this strain and age.
In order to study a possible contact allergenic potential of
Charcoal (Probe 1: C-Fix = 73.3%), three groups each of four female mice
were treated daily with the test item at concentrations of 2.5, 5, and
10% (w/w) in propylene glycol by topical application to the dorsum of
each ear (left and right) for three consecutive days. A control group of
four mice was treated with the vehicle (propylene glycol) only. Five
days after the first topical application the mice were injected
intravenously into a tail vein with 3HTdR. Approximately 5 h after
intravenous injection, the mice were sacrificed, the draining auricular
lymph nodes excised and pooled per group. Single cell suspensions of
lymph node cells were prepared from pooled lymph nodes, which were
subsequently washed and incubated with trichloroacetic acid overnight.
The proliferative capacity of pooled lymph node cells was determined by
the incorporation of 3HTdR measured in a beta-scintillation counter.
All treated animals survived the scheduled study period and no
signs of toxicity were observed.
A test item is regarded as a sensitiser in the LLNA if the
exposure to one or more test concentration resulted in 3-fold or greater
increase in incorporation of 3HTdR compared with concurrent controls, as
indicated by the Stimulation Index (S.I.). The estimated concentration
of test item required to produce a S.I. of 3 is referred to as the EC3
value. In this study Stimulation Indices of 0.65, 0.72, and 1.11 were
determined with the test item at concentrations of 2.5, 5, and 10%,
respectively, in propylene glycol. The EC3 value could not be
calculated, since none of the tested concentrations induced an S.I.
greater than 3.
The test item, charcoal (Probe 1: C-Fix = 73.3%) was not a skin
sensitiser under the test conditions of this study.
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