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Administrative data

Description of key information

Testing of the structural analogue d-limonene on rabbits demonstrated some moderate irritating effects on skin and eye. These irritation effects were not sufficiently severe for classification of l-limonene as skin irritating or eye irritating according to CLP criteria.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
no data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Read-across from an analogue substance for which there is a reliable in vivo study available (Klimish =2).
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See cross reference to justification of read-across.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
1.33
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
1.33
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritant / corrosive response data:
- Desquamation from skin surface was observed in all animals on Day 7
Interpretation of results:
GHS criteria not met
Remarks:
CLP Implementation.
Conclusions:
Based on the read-across from the analogue substance d-limonene, l-limonene is concluded to be irritating to the skin but the effects are not sufficient for classification according to CLP Regulation (EC) Nº 1272/2008.
Executive summary:

Based on the read-across from the analogue substance d-limonene, l-limonene is concluded to be irritating to the skin but the effects are not sufficient for classification according to CLP Regulation (EC) Nº 1272/2008.

Endpoint:
skin irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
no data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Read-across from an analogue substance for which there is a reliable in vivo study available (Klimish =2).
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See cross reference to justification of read-across.
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Irritation parameter:
erythema score
Basis:
mean
Remarks:
all animals
Time point:
other: 24, 48 and 72 h
Score:
1.92
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-acrossfrom the analogue substance d-limonene.
Irritation parameter:
edema score
Basis:
mean
Remarks:
all animals
Time point:
other: 24, 48 and 72 h
Score:
1.33
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Based on the read-acrossfrom the analogue substance d-limonene.
Other effects:
No data
Interpretation of results:
GHS criteria not met
Remarks:
CLP Implementation.
Conclusions:
Based on the read-across from the analogue substance d-limonene, l-limonene is concluded to be irritating to the skin but the effects are not sufficient for classification according to CLP Regulation (EC) Nº 1272/2008.
Executive summary:

Based on the read-across from the analogue substance d-limonene, l-limonene is concluded to be irritating to the skin but the effects are not sufficient for classification according to CLP Regulation (EC) Nº 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From May 10 to 21, 2010
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Read-across from an analogue substance for which there is a reliable in vivo study available (Klimish =2).
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See cross reference to justification of read-across.
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
conjunctivae score
Basis:
animal #1
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: 2 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 2 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
conjunctivae score
Basis:
animal #2
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 4 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 3 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
conjunctivae score
Basis:
animal #3
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
1.3
Max. score:
3
Reversibility:
fully reversible within: 4 days
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
mean individual score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: Based on the read-across from the analogue substance d-limonene.
Irritant / corrosive response data:
- Slight to moderate redness of conjunctivae associated with moderate chemosis was noted within 1 hour of instillation; irritation completely resolved within 7 days.
Other effects:
No data
Interpretation of results:
GHS criteria not met
Remarks:
CLP Implementation.
Conclusions:
Based on the read-across from the analogue substance d-limonene, l-limonene is concluded to be not irritating to the eyes and it is not classified according to CLP Regulation (EC) Nº 1272/2008.
Executive summary:

Based on the read-across from the analogue substance d-limonene, l-limonene is concluded to be not irritating to the eyes and it is not classified according to CLP Regulation (EC) Nº 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The only skin irritation study conducted with l-limonene (Moreno, 1975) was not sufficiently described for assessment (no detailled information and no scoring system). However the results of the study - moderate redness and moderate edema observed in 3/10 and 6/10 animals, respectively - were used as supportive information. A read-across approach with the structural analogue d-limonene was used to assess the irritation potential of l-limonene (see IUCLID § 13 for read-across justification).

In a primary dermal irritation study performed according to OECD guideline 404 and in compliance with GLP (ECETOCa, 1995), three albino rabbits were dermally exposed to d-limonene, under a semi-occlusive patch for 4 hours.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 2, 2, 2 for erythema score and 1.33, 2, 1.33 for oedema score. Signs of erythema and oedema, as well as desquamation from skin surface, were observed in all animals on Day 7.

Test was repeated under the same conditions. In this second experiment: mean individual scores at 24, 48 and 72 h after exposure for 4 animals were 2, 2, 1.67, 2 for erythema score and 1.33, 0.67, 1.33, 2 for oedema score. Slight desquamation was observed in 2/4 animals on day 7.

In an eye irritation study conducted according to the OECD Guideline 405 (ECETOCb, 1995), three male rabbits of the New Zealand White strain were exposed to d-limonene in one of the eyes while the other eye corresponded to the control.  

Instillation of d-limonene resulted in slight to moderate redness of conjunctivae. Mean individual scores at 24, 48 and 72 hours after exposure for the 3 animals were 0, 0, 0 for cornea score; 0, 0, 0 for iris score; 0.3, 1, 1.3 for conjunctivae score and 1, 0.3, 1 for chemosis score.

 

Effects on skin irritation/corrosion: slightly irritating

Effects on eye irritation: slightly irritating

Justification for classification or non-classification

According to the data available for skin irritation, l-limonene would not be classified according to CLP Regulation (EC) N° (1272-2008), as the maximum mean value in all animals was 2 for skin erythema which is lower than the classification criterion for the irritant category (at least 2 of 3 tested animals have a mean score of ≥ 2.3 - ≤ 4.0).

As only slight to moderate redness of conjunctivae (mean score < 2.0) was observed following rabbit testing with the anologue d-limonene for eye irritation, l-limonene is not classified as irritating to the eye according to the criteria of CLP Regulation (EC) N° (1272-2008).