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EC number: 227-815-6
CAS number: 5989-54-8
Several reliable studies were conducted on algae, daphnia and fish
with d-limonene, the enantiomer of l-limonene and with dipentene, the
racemic form (dl) of limonene. Toxicity results were similar for
d-limonene and dipentene suggesting that both enantiomer have the same
level of toxicity on these aquatic organisms. This hypothesis is
reinforced by the estimations of the toxicity of l-limonene by a valid
QSAR model (ECOSAR) which are similar to the result with d-limonene and
dipentene except for algae when the result is lower especially for
From these results, it can be concluded that the toxicity of the
two enantiomers, d and l limonene, regarding daphnids and fish is
similar and that d-limonene and dipentene can be used as analogue of
l-limonene in a read-across approach to fill data gaps. Among these
studies, the geometric mean 96h-LC50 of 0,71 mg/L for fish, the lowest
48h-EC50 of 0,36 mg/L and the 21d-NOEC= 0.27 mg/L for daphnia are
considered as the relevant values to be used for the purpose of risk
assessment and classification and labelling. Nevertheless, for algae the
lower estimated values, 72h-ErC and 72h-NOErC of 0.904 and 0.514
mg/L,for l-limonene with ECOSAR is prefered as more conservative values
because detailed information on the purity of d-limonene and dipentene
in the available tests is missing.
One reliable ready biodegradability study on l-limonene and one
reliable activated sludge respiration test (ASRIT) on dipentene (48.4%
dl-limonene) were available to assess the toxicity of l-limonene to
microorganisms. The result of the ASRIT on dipentene is difficult to
interpret the specific toxicity of l-limonene
(multi-constituents substance) however the test conclude that the test
item was not harmful to activated sludge. According to ECHA, Chapter R.
7b, Endpoint specific guidance (2008), the information content of the
ready biodegradability test can be used to derive a NOEC. In the case of
l-limonene, the ready biodegrability test in close bottles fulfilled the
requested conditions as described below :
- the compound is found to be readily biodegradable,
- test substance concentration (2 mg/L) was not above the
solubility limit of l-limonene (12.3 mg/L).
- a toxicity control hasn't been included in the test but no
inhibition of the endogenous respiration of the inoculum by l-limonene
Therefore, a NOEC of 2 mg/L corresponding to a good degradated
tested concentration was estimated from the biodegradation test and used
to derive the PNECstp.
PNEC sediment and soil
In the absence of any ecotoxicological data for sediment-dwelling
organisms and terrestrial organisms, the PNECsed and the PNECsoil were
calculated using the equilibrium partitioning method (EPM).
A 6-month subchronic repeated dose toxicity study was performed
similarly to OECD Guideline 409 with d-limonene administered through
gavage to groups of adult beagle dogs (IUCLID endpoint 7.5.1, RA,
5989-27-5, Repeated dose toxicity: oral 180-d dog, Webb, 1990, RS, K).
Under the test conditions, the NOAEL and LOAEL for beagle dogs
were considered to be 100 and 1000 mg/kg bw/day, respectively, based on
the increased absolute and relative female kidney weight and relative
male kidney weight.
NOEC mammals-food-chr of 4 g/ kg food is derived from the NOAEL and a
PNEC oral of 133 mg/kg food is calculated according to the ECHA guidance
on information requirments and chemical safety assessment Chapter 10
using a convertion factor
of 40 and an assessment factor of 30, respectively.
The selected data on acute and
chronic toxicity on aquatic organism issued from read-across with
d-limonene and dipentene or calculated value with a valid QSAR, and
relevant for classification of l-limonene are:
48h-EC50 for daphnia: 0.36
96h-LC50 for fish: 0.72 mg/L
72h-ECr50 for algae: 0.904 mg/l
21d-NOEC for daphnia: 0.27 mg/L
72h-NOErC for algae: 0.514 mg/L
The substance l-limonene is
The Log Kow of d-limonene was
measured at 4.38 and is expected to be similar for both enantiomers.
There was no experimentaly determined BCF however the retained QSAR
value is 683 L/kg ww and therefore l-limonene cannot be considered not
having potential to bioaccumulate.
Based on these information
l-limonene should be classified:
Acute Category 1 (M-factor = 1)
and Chronic Category 3 according to the CLP Regulation (EC) nº 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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