Hydrocarbons, terpene processing by-products

Endpoint summary

Description of key information

In an in vivo skin irritation study on the substance and in an in vitro skin irritation study on alpha pinene, results of irritation led to classification as irritating to skin, except in the in vivo study when considering CLP criteria.
In an in vivo eye irritation study in camphene and an in vitro study on Terpinolene multiconstituent, results of irritation led to classification as irritating to eye, except in the in vivo study when considering Directive 67/548/EEC criteria.

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In a primary dermal irritation study performed similarly to OECD Guideline 404 and in compliance with GLP, three New Zealand White rabbits were dermally exposed to 0.5 mL of undiluted test item, DERTOL 20 DD (Hydrocarbons, terpene processing by-products) under a semi-occlusive dressing for 4 h in normal and scarified skin. After removal of the dressings, irritation was scored as outlined in the OECD Guideline 404 at 1, 24, 48 and 72 h after exposure. Irritative phenomena were relatively important and slowly reversible. A slight oedema as well as slight erythema remained visible on the 6th day. Slight changes in skin structure were also observed. Mean scores over 24, 48 and 72 h for each animal were 2.0, 2.0 and 2.0 for erythema and 1.0, 1.0 and 1.7 for edema (normal skin) and 2.0, 2.0 and 2.0 for erythema and 1.0, 1.7 and 1.7 for edema (scarified skin). DERTOL 20 DD (Hydrocarbons, terpene processing by-products) is classified as "R38: irritating to skin" according to Directive 67/548/EEC and not classified as irritant according to CLP Regulation (EC) N° 1272/2008.

A GLP study conducted in vitro with human epidermis model EPISKIN was performed to assess the irritancy potential of alpha pinene (one of the main constituent of the substance) similarly to OECD guideline 439. 10 µL of test item was applied directly on epidermis for 15 min on 3 epidermis. Positive control was 10 µL of 5% (w/v) SDS solution and negative control was 10 µL of PBS (each tested on 3 epidermis). MTT conversion assay was peformed to evaluate the percentage of cellular viability of the epidermis. Positive control had a percentage of cell viability of 18.7 ± 3.0 and test item 39.6 ± 5.6. As the percentage of viability is ≤ 50 %, the test item is considered to be irritating to skin. Under the test conditions, alpha pinene is classified as irritating to skin according to the Directive 67/548/EEC and to the CLP Regulation (EC) N° 1272/2008.

 

In an in vitro eye irritation study performed in compliance with GLP, 30 µL of the test item, Terpinolene multiconstituent (a structure-related substance) was applied to the human reconstructed corneal epithelium model (in duplicate) for 10 ± 2 min, 1 h ± 10 min and 3 h ± 30 min at room temperature. Contact timepoint for negative control (sodium chloride 0.9 % w/v) was 3 h ± 30 min and 10 ± 2 min and 1 h ± 10 min for positive control (SDS 1.5% w/w). At the end of each incubation period, each epithelium was rinsed, transferred to new well containing MTT solution and incubated for 1 h ± 10 min in CO2 incubator. Each epithelium was then transferred into a new well plate containing isopropanol (1 mL). After agitating for 1 h ± 10 min, the extract (200μL/well) was transferred into a 96 wells microplate and optical density was recorded at 570 nm with a plates reader. For each treated tissue the viability was expressed as mean percentage of cellular viability relative to the negative control and the exposure time that causes 50% of cell mortality (T50) was determined by linear regression analysis. T50 value for the test item was 42.88 min. Optical density value for the negative control was 1.152 and T50 value for positive control was 33.49 min and thus confirmed the validity of the test. Under the test conditions, the T50 value of the test item was between 10 and 60 min therefore it was considered as moderately irritant when applied on a human reconstructed corneal epithelium.

In an eye irritation study conducted according to the OECD Guideline 405 and in compliance with GLP, three rabbits of the New Zealand White strain were exposed to 100 mg of undiluted camphene (one of the main constituent of the substance) in one of the eyes while the other eye corresponded to the control. The eyes were examined for irritation scores at 1 hour and 1, 2, 3, and 7 days after application. Examination and scoring after instillation of fluoresceine was assessed at 24 h, 72 h and 7 days after test substance application. Application of camphene resulted in severe redness of the conjunctivae associated with slight to moderate chemosis in all treated animals. The irritation completely resolved within 7 days. Mean individual scores at 24, 48 and 72 hours after exposure for the 3 animals were 1.3, 0.7, 0.7 for cornea score; 0.7, 0.7, 0.3 for iris score; 2.3, 3, 2.3 for conjunctivae score and 2, 1.7, 0.7 for chemosis score. Under the test conditions, camphene is classified as ‘H319 Cause severe eye irritation (Category 2)’ according to CLP Regulation (EC) N° 1272/2008 but not classified according to Directive 67/548/EEC.

Justification for selection of skin irritation / corrosion endpoint:
More than one study was available to assess the properties of the substance for this endpoint.

Justification for selection of eye irritation endpoint:
More than one study was available to assess the properties of the substance for this endpoint.

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

In an in vivo skin irritation study on the substance and in an in vitro skin irritation study on one of its main constituent alpha pinene, results of irritation led to classification as irritating to skin, except in the in vivo study with CLP criteria. Therefore, in a conservative approach, the substance is classified as R38 Irritating to skin according to the Directive 67/548/EEC and as Category 2, H315 Causes skin irritation according to the CLP Regulation (EC) No. 1272/2008.

In an in vivo eye irritation study on camphene, one of the main constituent of the substance, and an in vitro study on a structure-related substance Terpinolene multiconstituent, results of irritation led to classification as irritating to eye, except in the in vivo study with Directive 67/548/EEC criteria. Therefore, in a conservative approach, the substance was classified as R36 Irritating to eyes according to the Directive 67/548/EEC and Category 2 H319 Causes serious eye irritation according to CLP Regulation (EC) No. 1272/2008.