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EC number: 227-813-5
CAS number: 5989-27-5
- Dermal reactions: Application of
the test substance did not result in an increase in visual levels of
irritancy to the skin on or around the ear area for the duration of the
Table 1 - Skin sensitisation
potential of d-limonene in ethanol/diethyl phthalate (3:1 v/v)
Number of lymph nodes
per min (dpm)
per lymph node
0 (vehicle only)
Estimated to be 22% (5500 µg/cm2)
Table 2 - Skin sensitisation
potential of the positive control substance (hexylcinnamaldehyde)
Concentration of hexylcinnamaldehyde (% w/v)
Number of lymph nodes assayed
Disintegrations per min (dpm)
dpm per lymph node
In a local lymph node assay performed in
CBA/Ca strain mice according to OECD Guideline 429 and in compliance
with GLP, 25 µL of d-limonene were applied to groups of mice (4
females/dose) at concentrations of 0 (vehicle control), 10, 25, 50, 75
or 100% v/v in ethanol/diethyl phthalate (3: 1 v/v) to the dorsal
surface of each ear for three consecutive days. On Day 6, all animals
were injected with 3H-methyl thymidine and after five hours the draining
(auricular) lymph nodes were excised and measured for radioactivity
expressed as number of disintegrations per minute (DPM). Historic data
of hexylcinnamaldehyde (5, 10 and 25 % w/v) in acetone/olive oil (4:1
v/v) was used as the data for positive control group.
Mean DPM for 0, 10, 25, 50, 75 or 100%
d-limonene were observed to be 2511, 3319, 8554, 9916, 22063 or 16259
dpm, respectively. Stimulation index for 10, 25, 50, 75 or 100%
d-limonene were calculated to be 1.3, 3.4, 4.0, 8.8 or 6.5,
respectively. The estimated concentration giving rise to a 3 fold
increase in lymphocyte proliferation (EC3) was 22% v/v (5500 µg/cm2). No
increase in visual levels of irritancy to the ear skin was observed
during the study.
Therefore, d-limonene is classified as ‘R43
May cause sensitisation by skin contact’, according to Directive
67/548/EEC and in ‘Category 1’ according to CLP Regulation (EC) No
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