Lithium salt of branched-aliphatic dicarboxylic acid

Endpoint summary

Description of key information

 The irritation/corrosion effects of Lithium salt of branched-aliphatic dicarboxylic acid to skin and eyes were detected evaluated in reliable studies according to the appropriate standard test methods. For skin irritation, the study followed EU Method B.46 (in-vitro Skin Irritation: Reconstructed Human Epidermis Model Test) and OECD Guideline 439 (in-vitro Skin Irritation: Reconstructed Human Epidermis Model Test). For skin corrosion, the study followed EU Method B.40 (In Vitro Skin Corrosion: Human skin Model Test) and OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis). For eye irritation/corrosion, an in-vitro BCOP test according to EU Method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants) and OECD guideline 437 (Bovine corneal opacity and permeability test method) was conducted but allowed no prediction on the classification due to an IVIS between > 3 and 55. In consequence, a study following EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion), OECD Guideline 405 (Acute Eye Irritation / Corrosion) and EPA, OPPTS 870.2400 (1998) “Acute Eye Irritation” was conducted. Skin irritation: The relative mean tissue viability obtained after 15 minutes treatment with the test substance compared to the negative control tissues was 102%. Since the mean relative tissue viability for the test substance was above 50% after 15 minutes treatment, the test substance is considered to be non-irritant. Skin corrosion: The relative mean tissue viability obtained after 3-minute and 1-hour treatments with the test substance compared to the negative control tissues was 100% and 80%, respectively. Because the mean relative tissue viability for the test substance was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment, the test substance is considered to be not corrosive. Eye irritation: Eye iradial irritation with rabbits was observed 1 hour after application and resolved within 24 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 7 days in all animals. Based on these results, Lithium salt of aliphatic dicarboxylic acid does not have to be classified for eye irritation. 

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-08-19 to 2014-08-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

(July 26, 2013)

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Version / remarks:

(July 20, 2012)

GLP compliance:

yes

Species:

other: EPISKIN Small Model (TM)

Details on test animals or test system and environmental conditions:

TEST SYSTEM
- Test system: EPISKIN-SM (TM), 0.38 sq.cm, Lot no.: 14-EKIN-031
- Source: SkinEthic Laboratories, Lyon, France
- Acclimation period: preincubated for 26 h at 37°C in 12-well plates

MTT MEDIUM
- MTT concentrate 3 mg/mL in PBS diluted (10x) in Assay medium (final concentration 0.3 mg/mL)
- Source: Sigma Aldrich, Zwijndrecht, The Netherlands

ENVIRONMENTAL CONDITIONS DURING APPLICATION
- Temperature (°C): room temperature

ENVIRONMENTAL CONDITIONS DURING POST-TREATMENT INCUBATION
- Temperature (°C): 35.6-37.5 (continuously monitored)
- Humidity (%): 65-87 (continuously monitored)
- Other: CO2 content: 5.0 +/- 0.5% (monitored once daily on work days)

Type of coverage:

open

Preparation of test site:

other: no treatment

Vehicle:

unchanged (no vehicle)

Controls:

other: control tissues

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10.3 to 13. 0 mg

NEGATIVE CONTROL (NC)
- Substance: Phosphate buffered saline (PBS)
- Source: Invitrogen Corporation, Breda, The Netherlands
- Amount(s) applied (volume or weight with unit): 25 µL

POSITIVE CONTROL (PC)
- Substance: 5% (aq) Sodium dodecyl sulphate (SDS) in PBS
- Source: Sigma Aldrich, Zwijndrecht, The Netherlands)
- Amount(s) applied (volume or weight with unit): 25 µL and re-spread after 7 min contact time

Duration of treatment / exposure:

15 min

Observation period:

- Single obeservation after 45 h (Post-treatment incubation time 42 h; Incubation time in MTT-medium 3h)

Number of animals:

3 tissues each per test substance, negative and positive controls.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): all tissues were washed with PBS
- Time after start of exposure: 15 min
- Other: after rinsing all cell culture inserts were dried, moved to new well and incubated for 42 h at 37°C (post-treatment incubation)

OTHER: CELL VIABILITY MEASUREMENT
conducted after post-treatment incubation
- MTT incubation: cultured and dried inserts transferred to new plates with 2 mL MTT-medium (0.3 mg/mL) and incubated (3h at 37°C)
- Epidermis extraction: after MTT incubation tissues were dried on blotting paper; then epidermis was extracted with 500 µL isopropanol, then stored refrigerated in tubes for 72 h in darkness
- Formazan determination: spectrophotometrically at 570 nm in duplicate with the TECAN Infinite® M200 Pro Plate Reader

SCORING SYSTEM
- Cell viability: calculated for each tissue as percentage of the negative control mean
- Skin irritation potential: according to remaining cell viability

Irritation / corrosion parameter:

other: other: mean tissue viability

Value:

102

Remarks on result:

other:

Remarks:

Basis: mean. Remarks: 102 % of negative control. (migrated information)

Table 1 Mean absorption and tissue viability in the in vitro skin irritation test with Lithium salt of branched-aliphatic dicarboxylic acid

	A (OD570)	B (OD570)	C (OD570)	Mean (OD570)	SD	Mean tissue viability (percentage of control)
Negative control	1.069	1.057	1.047	1.058	+/- 0.011	100
Test substance	1.057	1.098	1.084	1.079	+/- 0.021	102
Positive control	0.371	0.245	0.657	0.425	+/- 0.211	40

 

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The relative mean tissue viability obtained after 15 minutes treatment with Lithium salt of branched-aliphatic dicarboxylic acid compared to the negative control tissues was 102%. Since the mean relative tissue viability for Lithium salt of branched-aliphatic dicarboxylic acid was above 50% after 15 minutes of treatment Lithium salt of branched-aliphatic dicarboxylic acid is considered to be non-irritant.

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:

experimental study

Adequacy of study:

key study

Study period:

2014-10-21 to 2014-11-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted under GLP conditions in accordance with the official OECD guideline No 405 (2012).

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

(2012)

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

(May, 2008)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

(August, 1998)

Qualifier:

according to guideline

Guideline:

other: JMAFF, 12 Nousan, Notification No 8147 (November, 2000; including the most recent partial revisions)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: between 12 and 24 weeks old
- Weight at study initiation: at least 1.5 kg
- Housing: individually housed in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits approx. 100 grams per day; hay and wooden sticks ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days before the start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): average 54.2 mg (range 54.0 - 54.4 mg)

Duration of treatment / exposure:

7 days

Observation period (in vivo):

Irritation: 1 hr, 24, 48, 72 hours and 7 days after instillation
Mortality/Viability: twice daily.
Toxicity: at least once daily.
Body Weight: day of treatment (prior to instillation) and after the final observation.

Number of animals or in vitro replicates:

3 males

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: according to OECD 405 and in addition:

Area of cornea involved:
No ulceration or opacity: 0
One quarter or less but not zero: 1
Greater than one quarter, but less than half: 2
Greater than half, but less than three quarters: 3
Greater than three quarters, up to whole area: 4

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals): 0
Any amount different from normal and/or lacrimation: 1
Discharge with moistening of the lids and hairs just adjacent to lids: 2
Discharge with moistening of the lids and hairs (considerable area around the eye): 3


TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

1

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

2

Reversibility:

fully reversible

Irritant / corrosive response data:

Irritation: Instillation of approximately 54 mg of Lithium salt of branched-aliphatic dicarboxylic acid (a volume of approximately 0.1 mL) into one eye of each of three rabbits resulted in effects on the iris and conjunctivae.
Iridial irritation was observed 1 hour after installation and resolved within 24 hours.
The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 7 days in all animals.

No corneal opacity was observed, and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.

Corrosion: There was no evidence of ocular corrosion.

Other effects:

Coloration / Remnants: No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.

Toxicity / Mortality: No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table1: Individual eye irritation scores

					Cornea				Iris		Conjunctivae				Comments
Animal	Time after dosing				Opacity (0-4)	Area (0-4)	Fluor area (%)^2		(0-2)		Redness (0-3)	Chemosis (0-4)	Discharge (0-3)
797^1	1 hour				0	0			1		2	2	2		-
	24 hours				0	0	0		0		2	1	1		-
	48 hours				0	0			0		1	1	1		-
	72 hours				0	0			0		1	0	0		-
	7 days				0	0			0		0	0	0		-
802	1 hour				0	0			1		2	2	2		-
	24 hours				0	0	0		0		2	1	1		-
	48 hours				0	0			0		1	0	1		-
	72 hours				0	0			0		1	0	1		-
	7 days				0	0			0		0	0	0		-
805	1 hour				0	0			1		2	2	2		-
	24 hours				0	0	0		0		2	1	1		-
	48 hours				0	0			0		1	1	1		-
	72 hours				0	0			0		1	1	1		-
	7 days				0	0			0		0	0	0		-

 

¹Sentinel,²  Green staining after fluorescein treatment (percentage of total corneal area)

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Iridial irritation was observed 1 hour after installation and resolved within 24 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 7 days in all animals. Based on these results, Lithium salt of branched-aliphatic dicarboxylic acid does not have to be classified for eye irritation.

Executive summary:

Acute eye irritation/corrosion study with Lithium salt of branched-aliphatic dicarboxylic acid in the rabbit.

 

The study was carried out based on the guidelines described in:

OECD No.405 (2012) "Acute Eye Irritation / Corrosion"

EC, No 440/2008, B5: "Acute Toxicity: Eye Irritation/Corrosion"

EPA, OPPTS 870.2400 (1998): "Acute Eye Irritation"

JMAFF Guidelines (2000), including the most recent revisions.

 

Single samples of approximately 54 mg of Lithium salt of branched-aliphatic dicarboxylic acid (a volume of approximately 0.1 mL) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and 7 days after instillation.

 

Instillation of the test substance resulted in effects on the iris and conjunctivae. Iridial irritation was observed 1 hour after installation and resolved within 24 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 7 days in all animals.

  

Based on these results, Lithium salt of branched-aliphatic dicarboxylic acid does not have to be classified and has no obligatory labelling requirement for eye irritation according to the:

- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011)(including all amendments),

- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:
Two studies are available. The study for skin irritation was conducted under GLP conditions according to OECD guideline 439 and matched the requirements of Annex 7 of the REACH legislation. The study for skin corrosion was also conducted under GLP conditions according to OECD guideline 431.

Justification for selection of eye irritation endpoint:
In a BCOP study for EYE IRRITANCY POTENTIAL OF LITHIUM SALT OF ALIPHATIC DICARBOXYLIC ACID according to OECD guideline 437, the in vitro irritancy score (INIS) of 7.5 after 240 minutes of treatment was determined. Since Lithium salt of branched-aliphatic dicarboxylic acid induced an IVIS between > 3 and 55, no prediction on the classification could be made. A in vivo test had to be conducted to conclude on the classification.

The study was conducted under GLP conditions according to OECD guideline 405 and matched the requirements of Annex 7 of the REACH legislation.

Justification for classification or non-classification

All three studies, skin irritation, skin corrosion and in-vivo eye irritation, show no adverse effects.