Polar modified Rice Bran Wax

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27-Jul-2015 to 03-Aug-2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Guideline no. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10.0-12.0 mg, moistened with 5 µl water

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 10 µl Phosphate buffered saline
POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 10 µl
- Concentration (if solution): 5% (aq) Sodium dodecyl sulphate

Duration of treatment / exposure:

Exposure:15 minutes
Post incubation period: 42 hours

Details on study design:

TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Irritation / corrosion parameter:

other: other: percentage viability

Value:

72

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 15 minutes. (migrated information)

Interpretation of results:

not irritating

Remarks:

Migrated information

Conclusions:

The in vitro skin irritation test was conducted according to OECD 439 guideline and GLP principles.

It is concluded that this test is valid and that Licocare RBW 106 is non-irritant in the in vitro skin irritation test.

Executive summary:

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with Licocare RBW 106 compared to the negative control tissues was 72%. Since the mean relative tissue viability for Licocare RBW 106 was above 50% after 15 minutes treatment it is considered to be non-irritant.

 

The positive control had a mean cell viability of 8% after 15 minutes exposure. The absolute mean OD₅₇₀of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 7%, indicating that the test system functioned properly.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 March 2020 to 27 March 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

Section 4: Health Effects
adopted on 28 July 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Clariant Plastics and Coatings (Deutschland) GmbH and DEF2105728
- Expiration date of the lot/batch: 16.01.2022



STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29°C)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Adita Biosys Private Limited
- Age at study initiation: 5 Months
- Weight at study initiation: 2.16531 kg to 2.27551 kg
- Housing: stainless steel wire mesh cage
- Diet (e.g. ad libitum): yes
- Water (e.g. ad libitum): yes
- Acclimation period: 12 March 2020 to 23 March 2020

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5°C to 22.8°C
- Humidity (%): 46% to 66%
- Air changes (per hr): 12 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark and 12 hours light

IN-LIFE DATES: From: 17 March 2020 To: 27 March 2020

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g


VEHICLE
- Amount(s) applied (volume or weight with unit): 1 mL of distilled water
- Lot/batch no. (if required): 8721 and 8730

Duration of treatment / exposure:

Initial Test: 3 min, 1 hour, 4 hour
Confirmatory Test: 4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: Trunk region
- % coverage: approximately 6 cm2
- Type of wrap if used: crepe bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 3 min, 1 hour and 4 hour for inital test and 4 hours for confirmatory test

OBSERVATION TIME POINTS
3 min, 1 hour, 4 hour, 24 hour, 48 hour and 72 hour in initial test
1 hour, 24, 48 and 72 hour in confirmatory test

SCORING SYSTEM:
- Method of calculation: mean scoring

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: no irritation

Remarks:

No irritation observed

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: .

Remarks:

no effect observed

Remarks on result:

no indication of irritation

TABLE 1.     INDIVIDUAL ANIMAL CLINICAL SIGNS OF TOXICITY AND MORTALITY RECORD

Phase of the Experiment	Dose (g/Patch)	Animal No.	Sex	Clinical Signs of Toxicity and Mortality on Day
					1	2	3	4
Initial Test	0.5	Nb1728	Male	N	N	N	N
Confirmatory Test	0.5	Nb1729	Male	N	N	N	N
			Nb1730	Male	N	N	N	N

N: Normal

TABLE 2.     INDIVIDUAL ANIMAL SKIN REACTIONS SCORING RECORD

Phase of the Experiment	Animal No.	Sex	Exposure   Duration	Application Site	Dose (g/Patch)	Observations	Skin Reactions Score after Patch Removal					Mean Score
							IAR	1 hr	24 hrs	48 hrs	72 hrs
Initial Test	Nb1728	Male	3 min	Site 2	0.5	ERY	0	-	-	-	-	-
						EDE	0	-	-	-	-	-
			1 hr	Site 3	0.5	ERY	0	-	-	-	-	-
						EDE	0	-	-	-	-	-
			4 hrs	Site 1	Untreated Control	ERY	0	0	0	0	0	0
						EDE	0	0	0	0	0	0
			4 hrs	Site 4	0.5	ERY	0	0	0	0	0	0
						EDE	0	0	0	0	0	0
Confirmatory Test	Nb1729	Male	4 hrs	Site 1	Untreated Control	ERY	-	0	0	0	0	0
						EDE	-	0	0	0	0	0
				Site 2	0.5	ERY	-	0	0	0	0	0
						EDE	-	0	0	0	0	0
	Nb1730	Male	4 hrs	Site 1	Untreated Control	ERY	-	0	0	0	0	0
						EDE	-	0	0	0	0	0
				Site 2	0.5	ERY	-	0	0	0	0	0
						EDE	-	0	0	0	0	0

 ERY: Erythema; EDE: Oedema; 0: No Erythema/Oedema; min: Minutes; hr/hrs: Hour/Hours; IAR: Immediately after removal of the test patch; -: Not applicable

 

                                  

Mean Score For Erythema/ Oedema	:	24 hrs + 48 hrs+ 72 hrs (Observations after patch removal)
		3 (Observation time point)

 

TABLE 3.     BODY WEIGHT (kg) AND PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1

Phase of the Experiment	Dose (g/Patch)	Animal No.	Sex	Body Weight (kg) on day		Percent Change in Body Weight with Respect to Day
				1	4	1 to 4
Initial Test	0.5	Nb1728	Male	2.22915	2.26567	1.63829
Confirmatory Test	0.5	Nb1729	Male	2.28431	2.33922	2.40379
	0.5	Nb1730	Male	2.32411	2.36998	1.97366
		Mean		2.30421	2.35460	2.18872
		±SD		0.02814	0.02175	0.30415
		n		2	2	2

SD: Standard Deviation; n: Number of animals

 

 

TABLE 4.     INDIVIDUAL ANIMAL GROSS PATHOLOGY FINDINGS

Phase of the Experiment	Dose (g/Patch)	Animal No.	Sex	Fate	Gross Pathology Findings
					External	Internal
Initial Test	0.5	Nb1728	Male	TS	NAD	NAD
Confirmatory Test	0.5	Nb1729	Male	TS	NAD	NAD
		Nb1730	Male	TS	NAD	NAD

NAD: No Abnormality Detected; TS: Terminal Sacrifice

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the experiment, it is concluded that single dermal application of Licocare RBW 106 TP did not reveal any skin reactions when applied to the skin of New Zealand White Rabbits, hence the test item is considered as non-irritant and did not meet the classification criteria according to the Globally Harmonized System (GHS) of Classification and Labelling of Chemicals

Executive summary:

The test item, Licocare RBW 106 TPwas evaluated forAcute Dermal Irritation/Corrosion in New Zealand White rabbits as per OECD Guidelines for Testing of Chemicals No. 404 (Section 4: Health Effects), “Acute Dermal Irritation/Corrosion” adopted on 28 July 2015.

The study was performed in two phases i.e. initial test and confirmatory test.            Approximately 24 hours before the application of the test item, fur on the                    dorso-lateral area of the trunk of the animals was removed by clipping closely using an electric hair clipper (approximately 8 × 12 cm). Care was taken to avoid abrasion to the skin and the animals with healthy intact skin were used for experiment.

As there was no information on the test item, in initial test, three patches were applied for 3 minutes, 1 hour and 4 hour exposure period. The application site 1 (anterior left trunk region) of the animal was served as untreated control. On application site 2 (posterior left trunk region), site 3 (posterior right trunk region) and site 4 (anterior right trunk region) of the animal, 0.5 g oftest item moistened with 1.0 mL of distilled water was applied for 3 minutes, 1 hour and 4 hours respectively (semi-occlusive dressing). The first test patch was applied on site 2 and removed after three minutes. No skin reaction was observed after immediate observation, a second test patch was applied at site 3 and removed after one hour. After immediate observation, the observations at this stage indicated that exposure could be humanely allowed to extend to four hours, a third test patch was applied at site 4 and removed after four hours, and the response was graded. The application site 1 (anterior left trunk region) of the animal was covered with blank cotton gauze at the time of site 4 application and was removed along with application site 4. After removal of the test patches, the application sites were washed with distilled water and dried with absorbent cotton without altering the existing response or the integrity of the epidermis. The application sites 1, 2, 3 and 4 were scored immediately after patch removal whereas the application sites 1 and 4 were also scored approximately at 1, 24, 48 and 72 hours after the patch removal.

As there was no corrosive effects observed during initial test, the confirmatory test was performed using two additional rabbits. In the confirmatory test, the patches were applied for 4 hours exposure period. Each animal had two application sites. The clipped area of skin at left trunk region and right trunk region were identified as application sites 1 and 2 respectively.

In the confirmatory test, the application site 1 (left trunk region) of each animal was served as untreated control covered with blank cotton gauze. On application site 2 (right trunk region) of each animal, 0.5 g oftest item moistened with 1.0 mL of distilled water was applied for 4 hours exposure period and removed after 4 hours. After removal of the test patches, the application sites were washed with distilled water and dried with absorbent cotton without altering the existing response or the integrity of the epidermis. The application sites 1 and 2 were scored approximately at 1, 24, 48 and             72 hours after patch removal.

The mean score across 3 scoring times (24, 48 and 72 hours after patch removal) for both initial test animal and confirmatory test animals was ‘0’ for erythema and                 oedema grades.

No erythema and oedema was observed at the test item application site in both initial and confirmatory test animals.

All the animals were observed once daily for clinical signs of toxicity and twice daily for mortality during the observation period. No clinical signs of toxicity and mortality was observed in both initial and confirmatory test animals.

Body weight was recorded on the day of receipt and prior to application of test item (day 1) and at termination. No changes were observed in body weight andpercent change in body weight with respect to day 1in both initial and confirmatory test animals. All the animals revealednormal increase in the body weightduring the observation period.

At termination, all the animals were sacrificed by intravenous administration of sodium thiopentone. Nogross pathological findings were observed in any of the animals in both initial and confirmatory test

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30-Jul-2015 to 11-Aug-2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

other: OECD guideline 437 “Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage”

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EU Method B.47 “Bovine corneal opacity and permeability method for identifying ocular corrosives and severe irritants

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approx. 300 mg per cornea

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount(s) applied (volume or weight with unit): 750 µl per cornea
Concentration (if solution): 20% (w/v) Imidazole

Duration of treatment / exposure:

240 minutes

Details on study design:

TEST SITE
- Isolated bovine cornea

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 240 minutes

SCORING SYSTEM:
- After exposure the cornea is thoroughly rinsed to remove the test substance followed by immediate opacity measurement and permeability evaluation of the cornea.

- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:

In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).


TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

DATA EVALUATION:
In vitro score range UN GHS
≤ 3 No Category
> 3; ≤ 55 No prediction can be made
>55 Category 1

Irritation parameter:

in vitro irritation score

Run / experiment:

means

Value:

2.8

Negative controls validity:

valid

Positive controls validity:

valid

The individual in vitro irritancy scores for the negative controls ranged from -0.7 to 0.5. The individual positive control in vitro irritancy scores ranged from 146 to 156. The corneas treated with the positive control were turbid after the 240 minutes of treatment.

The corneas treated with Licocare RBW 106 showed opacity values ranging from 1.8 to 3.8 and permeability values ranging from -0.008 to 0.011. The corneas were slightly translucent after the 240 minutes of treatment with Licocare RBW 106. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from 2.0 to 3.7 after 240 minutes of treatment with Licocare RBW 106.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Since Licocare RBW 106 induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

Executive summary:

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas.The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 150 and within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

 

Licocare RBW 106 did not induce ocular irritation through both endpoints, resulting in a meanin vitroirritancy score of 2.8 after 240 minutes of treatment.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 February 2020 to 12 March 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Section 4: Health Effects
adopted on 09 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Clariant Plastics and Coatings (Deutschland) GmbH and DEF2105728
- Expiration date of the lot/batch: 16.01.2022



STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29°C)

FORM AS APPLIED IN THE TEST (if different from that of starting material) Not applicable

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Adita Biosys Private Limited
- Age at study initiation: 4 Months
- Weight at study initiation: 2.13264 kg to 2.31243 kg
- Housing: individually in stainless steel wire mesh cage
- Diet (e.g. ad libitum): Yes
- Water (e.g. ad libitum): Yes
- Acclimation period: 25 February 2020 to 08 March 2020

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4°C to 22.8°C
- Humidity (%): 45% to 67%
- Air changes (per hr): 12 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark and 12 hrs light

IN-LIFE DATES: From: 02 March 2020 To: 12 March 2020

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

1 hour from test item instillation

Observation period (in vivo):

72 hours

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 1 hour post exposure

SCORING SYSTEM: 1 hour, 24, 48 and 72 hour

TOOL USED TO ASSESS SCORE: hand-slit lamp / fluorescein

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other:

Remarks:

no effect observed

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other:

Remarks:

no effect observed

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other:

Remarks:

no effect observed

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other:

Remarks:

no effect observed

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Initial Test Sex: Male Dose: 0.1 mL/animal Animal No.: Nb1581
Observation Period Ocular Lesions
Conjunctiva Iris Cornea
Redness Chemosis Opacity Area
Eyes LE RE LE RE LE RE LE RE LE RE
1 hr 1 0 0 0 0 0 0 0 - -
24 hrs 0 0 0 0 0 0 0 0 - -
48 hrs 0 0 0 0 0 0 0 0 - -
72 hrs 0 0 0 0 0 0 0 0 - -
Mean Tissue Score 0 0 0 0 0 0 0 0 - -


Confirmatory Test Sex: Male Dose: 0.1 mL/animal Animal No.: Nb1582
Observation Period Ocular Lesions
Conjunctiva Iris Cornea
Redness Chemosis Opacity Area
Eyes LE RE LE RE LE RE LE RE LE RE
1 hr 1 0 0 0 0 0 0 0 - -
24 hrs 0 0 0 0 0 0 0 0 - -
48 hrs 0 0 0 0 0 0 0 0 - -
72 hrs 0 0 0 0 0 0 0 0 - -
Mean Tissue Score 0 0 0 0 0 0 0 0 - -


Confirmatory Test Sex: Male Dose: 0.1 mL/animal Animal No.: Nb1583
Observation Period Ocular Lesions
Conjunctiva Iris Cornea
Redness Chemosis Opacity Area
Eyes LE RE LE RE LE RE LE RE LE RE
1 hr 1 0 0 0 0 0 0 0 - -
24 hrs 0 0 0 0 0 0 0 0 - -
48 hrs 0 0 0 0 0 0 0 0 - -
72 hrs 0 0 0 0 0 0 0 0 - -
Mean Tissue Score 0 0 0 0 0 0 0 0 - -
hr: hour; hrs: hours; LE: Left Eye (Treated Eye); RE: Right Eye (Untreated/Reference control Eye)
Mean Tissue Score = (24 hr+48 hr+72 hr)/3
Redness: 0: Normal; 1: Some blood vessels hyperaemic (injected)
Chemosis: 0: Normal
Iris: 0: Normal
Opacity: 0: No ulceration or opacity

Other effects:

- Lesions and clinical observations: no lesions or clinical signs observed
- Ophthalmoscopic findings: no reactions
- Histopathological findings: no abnormalities detected
- Effects of rinsing or washing: no effects

             TABLE 1. INDIVIDUAL ANIMAL CLINICAL SIGNS OF TOXICITY AND MORTALITY RECORD

Phase of the Experiment	Dose (mL/animal)	Animal No.	Sex	Clinical Signs of Toxicity and Mortality on Day
				1		2		3		4
				#	*	#	*	#	*	#
Initial Test	0.1	Nb1581	M	N	N	N	N	N	N	N
Confirmatory Test	0.1	Nb1582	M	N	N	N	N	N	N	N
	0.1	Nb1583	M	N	N	N	N	N	N	N

           M: Male; N: Normal; #: First observation; *: Second observation

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                                                                                   

                                                                                   

            TABLE 2. INDIVIDUAL ANIMAL EYE IRRITATION/CORROSION SCORING RECORD

Initial Test                           Sex: Male                     Dose: 0.1 mL/animal                   Animal No.:Nb1581
Observation Period		Ocular Lesions
		Conjunctiva					Iris			Cornea
		Redness			Chemosis					Opacity			Area
Eyes		LE		RE		LE		RE		LE		RE		LE		RE		LE		RE
1 hr		1		0		0		0		0		0		0		0		-		-
24 hrs		0		0		0		0		0		0		0		0		-		-
48 hrs		0		0		0		0		0		0		0		0		-		-
72 hrs		0		0		0		0		0		0		0		0		-		-
Mean Tissue Score		0		0		0		0		0		0		0		0		-		-

hr: hour; hrs: hours; LE: Left Eye (Treated Eye); RE: Right Eye (Untreated/Reference control Eye);

Mean Tissue Score = (24 hr+48 hr+72 hr)/3

Redness: 0: Normal; 1: Some blood vessels hyperaemic (injected)

Chemosis:0: Normal

Iris:0: Normal               

Opacity:0: No ulceration or opacity

 

 

 

 

 

 

 

 

 

 

TABLE 2 (Contd...).  INDIVIDUAL ANIMAL EYE IRRITATION/CORROSION SCORING RECORD

Confirmatory Test                           Sex: Male                     Dose: 0.1 mL/animal      Animal No.:Nb1582
Observation Period	Ocular Lesions
	Conjunctiva				Iris		Cornea
	Redness		Chemosis				Opacity		Area
Eyes	LE	RE	LE	RE	LE	RE	LE	RE	LE	RE
1 hr	1	0	0	0	0	0	0	0	-	-
24 hrs	0	0	0	0	0	0	0	0	-	-
48 hrs	0	0	0	0	0	0	0	0	-	-
72 hrs	0	0	0	0	0	0	0	0	-	-
Mean Tissue Score	0	0	0	0	0	0	0	0	-	-

 

 

Confirmatory Test                           Sex: Male                     Dose: 0.1 mL/animal      Animal No.:Nb1583
Observation Period	Ocular Lesions
	Conjunctiva				Iris		Cornea
	Redness		Chemosis				Opacity		Area
Eyes	LE	RE	LE	RE	LE	RE	LE	RE	LE	RE
1 hr	1	0	0	0	0	0	0	0	-	-
24 hrs	0	0	0	0	0	0	0	0	-	-
48 hrs	0	0	0	0	0	0	0	0	-	-
72 hrs	0	0	0	0	0	0	0	0	-	-
Mean Tissue Score	0	0	0	0	0	0	0	0	-	-

hr: hour; hrs: hours; LE: Left Eye (Treated Eye); RE: Right Eye (Untreated/Reference control Eye)

Mean Tissue Score = (24 hr+48 hr+72 hr)/3

Redness: 0: Normal; 1: Some blood vessels hyperaemic (injected)

Chemosis:0: Normal

Iris:0: Normal               

Opacity:0: No ulceration or opacity

 

 

 

 

 

 

 

 

 

                                      TABLE 3.  INDIVIDUAL ANIMAL SLIT LAMP EXAMINATION RECORD

Initial Test                   Sex: Male                       Dose: 0.1 mL/animal                         Animal No.: Nb1581
	Day 2 (24 hours)
Eye	LE	RE
Lids	N	N
Ducts	N	N
Cornea	N	N
Pupil	N	N
Sclera	N	N
Ciliary Bodies	N	N
Iris	N	N
Aqueous Humour	N	N
Lens	N	N
Vitreous Humour	N	N

LE: Left Eye; RE: Right Eye; N: Normal/No Abnormality Detected

 

 

 

 

 

 

 

TABLE 3 (Contd…). INDIVIDUAL ANIMAL SLIT LAMP EXAMINATION RECORD

Confirmatory Test                   Sex: Male                       Dose: 0.1 mL/animal            Animal No.: Nb1582
	Day 2 (24 hours)
Eye	LE	RE
Lids	N	N
Ducts	N	N
Cornea	N	N
Pupil	N	N
Sclera	N	N
Ciliary Bodies	N	N
Iris	N	N
Aqueous Humour	N	N
Lens	N	N
Vitreous Humour	N	N

 

 

Confirmatory Test                   Sex: Male                       Dose: 0.1 mL/animal            Animal No.: Nb1583
	Day 2 (24 hours)
Eye	LE	RE
Lids	N	N
Ducts	N	N
Cornea	N	N
Pupil	N	N
Sclera	N	N
Ciliary Bodies	N	N
Iris	N	N
Aqueous Humour	N	N
Lens	N	N
Vitreous Humour	N	N

LE: Left Eye; RE: Right Eye; N: Normal/No Abnormality Detected

 

 

 

   TABLE 4.   INDIVIDUAL ANIMAL BODY WEIGHT (kg) AND PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1

Phase of the Experiment	Dose (mL/animal)	Animal No.	Sex	Body Weight (kg) on Day		Percent Change in Body Weight with Respect to Day
				1	4	1 to 4
Initial Test	0.1	Nb1581	Male	2.34193	2.37243	1.30234
Confirmatory Test	0.1	Nb1582	Male	2.45906	2.48941	1.23421
	0.1	Nb1583	Male	2.27159	2.31234	1.79390
			Mean	2.36533	2.40088	1.51405
			±SD	0.13256	0.12521	0.39576
			n	2	2	2

 SD: Standard deviation; n: Number of animals

 

 

 

      

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                                                TABLE 5.  INDIVIDUAL ANIMAL GROSS PATHOLOGYFINDINGS

Phase of the Experiment	Dose (mL/animal)	Animal No.	Sex	Fate	Gross Pathology Findings
					External	Internal
Initial Test	0.1	Nb1581	Male	TS	NAD	NAD
Confirmatory Test	0.1	Nb1582	Male	TS	NAD	NAD
	0.1	Nb1583	Male	TS	NAD	NAD

TS: Terminal Sacrifice; NAD: No Abnormality Detected

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the above results of the experiment and under the experimental conditions employed, it is concluded that the test item Licocare RBW 106 TP was “non-irritant” to the rabbit eye following single ocular instillation. Hence, the test item is unclassified as per the Globally Harmonized System of Classification and Labelling of Chemicals (GHS)

Executive summary:

The test item,Licocare RBW 106 TPwas evaluated for Acute Eye Irritation/Corrosion in New Zealand White Rabbitsas perOECD 405 (Section 4: Health Effects), “Acute Eye Irritation/Corrosion” adopted on 09 October 2017.The study was performed in two phases i.e., initial and confirmatory tests. Both the eyes of each experimental animal were examined within 24 hours before the treatment. The animals with absence of signs of eye irritation, ocular defects or pre-existing corneal injury were used for the treatment. The pH of test item is 6.44 as per in-house pH determination.The initial test was conducted using single male rabbit and confirmatory test was conducted using two male rabbits. The systemic analgesic (tramadol hydrochloride injection 2.5 mg/kg) and two drops of topical ocular anaesthetic (0.5% tetracaine hydrochloride) were given to each animal sixty minutes and five minutes prior to test item instillation respectively. The 0.1 mL (Initial test: 69.50 mg and Confirmatory test: 65.93 mg and 64.02 mg) of test item was instilled into the conjunctival sac of the left eye and right eye served as control. The eyes were scored approximately at 1, 24, 48 and 72 hours. Slit lamp examination was carried out using fluorescein strips (fluorescein sodium ophthalmic strip USP) and were scored approximately at                  24 hours for both initial and confirmatory test.All the animals (initial test and confirmatory test) were observed twice daily for clinical signs of toxicity and mortality. No treatment related clinical signs of toxicity and mortality were observed in all the animals (initial and confirmatory tests) after the test item instillation.In both initial and confirmatory test, treated eye (left) revealed ocular lesions like redness at 1 hour observation. The observed lesion reversed back to normal by          24 hour observation.In both initial and confirmatory test, no ocular lesions were observed at 24 hour observation during slit lamp examination.The body weight was recorded on the day of receipt, on the day of treatment (prior to instillation of test item) and on the day of termination of the experiment. No changes were noted in body weight and percent change in body weight with respect to day 1 in both initial and confirmatory test. All the animals revealed physiologically normal increase in body weight in both initial and confirmatory tests.All the animals were sacrificed by intravenous administration of sodium thiopentone and observed for gross pathological changes. Nogross pathological findings were observed in all the animals.

For initial test and confirmatory test, the mean score calculated across 3 scoring times (approximately 24, 48 and 72 hours after test item

instillation) for cornea, iris and conjunctival redness and conjunctival chemosis were 0, 0, 0 and 0 respectively

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available