p-nitrobenzoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)

Endpoint summary

Description of key information

Skin irritation:

In an in vivo skin irritation study in New Zealand White rabbits according to OECD Guideline 404 and EU Method B.4 (Klimisch 2, Safepharm, 2001), p-Nitrobenzoic acid, compound with 2,2',2''-nitrilotriethanol (1:1) was observed to be not corrosive and not irritating to skin. The study is assigned as Klimisch 2 since detailed information on the testing procedure is missing in the report.

Eye irritation:

In a K1 acute eye irritation study (Toxicoop, 2017) in New Zealand White rabbits according to the Draize method (Draize, 1959; OECD Guideline 405, 2012), p-Nitrobenzoic acid, compound with 2,2’,2’’-nitrilotriethanol (1:1) was not considered to be an eye irritant. The test item applied to the rabbits' eye mucosa caused slight conjunctival irritant effects which were fully reversible within 1 week.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 - 14 October 2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The study can be used to draw a conclusion on the non corrosive/ non-irritating properties to skin of the tested substance. The report consists of a summary of the conducted study including detailed raw data without detailed information on the procedure followed.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

semiocclusive

Preparation of test site:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 ml

VEHICLE
- not required

Duration of treatment / exposure:

4 hour

Observation period:

1, 24, 48 and 72 hours after administration

Number of animals:

3

Details on study design:

TEST SITE
the test material was administered to the intact skin

OBSERVATION TIME POINTS
1, 24, 48 and 72 hours after administration.

SCORING SYSTEM:
- Method of calculation: prinmary irritation index, according to Draize (1959)

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

animal #3

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

To evaluate the classification of the substances according to CLP Regulation, the raw data were reinterpreted.
The mean value from gradings at 24, 48 and 72 hours after patch removal calculated in at least 2 of 3
tested animals were the following:
- erythema/eschar: score 0/6=0 (3 out of 3 animals)
- oedema: 0/6=0 (3 out of 3 animals)

Other effects:

None reported

Interpretation of results:

GHS criteria not met

Conclusions:

The results allows clear decision that the substance p-Nitrobenzoic acid, compound with 2,2',2''-nitrilotriethanol (1:1) is not corrosive and not irritating to skin according to the regulation 1272/2008/EC after reinterpretation of the raw data.

Executive summary:

A study was performed to assess the irritation potential of the test material p-Nitrobenzoic acid, compound with 2,2',2''-nitrilotriethanol (1:1) to the skin of the New Zealand White rabbit (SPL Standard Test Method 540.06). The method followed OECD Guidelines for Testing of Chemicals (1992) No. 404 "Acute Dermal lrritation/Corrosion" and Method 84 of Commission Directive 92l69lEEC (which constitutes Annex V of Council Directive 67l54SlEEC).

The results may be used as a basis for classification and labelling according to the Regulation 1272/2008/EC after reinterpretation of the raw data.

A single 4-hour semi-occluded application (0.5 ml) of the test material was administered to the intact skin of three rabbits. Skin reactions were recorded 1, 24, 48 and 72 hours after administration.

The mean value from gradings at 24, 48 and 72 hours after patch removal calculated were the following:

- erythema/eschar: score 0/6=0 (3 out of 3 animals)

- oedema: 0/6=0 (3 out of 3 animals)

No evidence of skin irritation/corrosion was noted. The test material is not corrosive and not irritant to skin according to the regulation 1272/2008/EC.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

Reason / purpose for cross-reference:

reference to other study

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:

May 03 - June 14, 2017

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:

2nd October 2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Source: S & K-LAP Kft. 2173 Kartal, Császár út 135, HUNGARY
Hygienic level during the study: good conventional
Justification of strain: The New Zealand white rabbit is one of the standard animals in acute eye irritation study.
Number of animal: 3 males
Age of animals: adult rabbits, 12 weeks old
Body weight range at the beginning of the study: 2560 - 2618 g
Body weight range at the end of the study: 2850 - 2898 g
Date of receipt: April 28, 2017
Acclimatisation time: 5 days in first animal, 6 days in second animal and 7 days in third animal

Animal health: Only animals in acceptable health condition were used for the test.
Room: 11/C
Housing: Animals were housed individually in metal cage.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20 ± 3 °C
Relative humidity: 30-70 %
Ventilation: above 10 air exchanges/hour by central air-condition system
The environmental parameters were recorded daily during the study.
Food: The animals received S&K LAP rabbit diet produced by Cargill Takarmány Zrt., 5300 Karcag, Madarasi út 0399. Hungary, ad libitum.
Water supply: Animals received tap water from watering bottles ad libitum.
The drinking water is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Identification: Individual animal identification of the rabbits was performed by numbered ear tag.
The cages were marked with individual identity cards with information about study number, test item, sex, strain, dose group, date of arriving of animals, room number, cage number and individual animal numbers.

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1g of the test item was used.

Duration of treatment / exposure:

The eyes of the test animals were not washed out 24 hour after test item application, because the test item had been removed from the eye of test animal by physiological mechanisms.

Observation period (in vivo):

The eyes were examined at 1, 24, 48 and 72 hours then 1 week after the application. The duration of the observation period was sufficient for the statement of reversibility or irreversibility of changes.
Any clinical signs of toxicity or signs of pain and/or distress were recorded at least twice daily, with a minimum of 6 hours between observations.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

The eye irritation scores were evaluated according to the scoring system by Draize (1979) and OECD 405 (02 October 2012) presented by the following table.
1. Conjunctivae
A. Redness (Palpebral and bulbar)
Normal 0
Some blood vessels hyperaemic (injected) 1
Diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3

B.Chemosis
Normal 0
Some swelling above normal (includes nictating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

C. Discharge

No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, on considerable area
around the eye 3
2. Iris
D. Values
Normal 0
Markedly deepened rugae, congestion, swelling, moderate
circumcorneal hyperaemia: or injection: iris reactive to light
(a sluggish reaction is considered to be an effect) 1
Hemorrhage, gross destruction, or no reaction to light. 2

3. Cornea
E. Opacity-degree of density (Area most dense taken for reading)
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight
dulling of normal lustre): details of iris clearly visible 1
Easily discernible translucent area: details of iris slightly obscured 2
Nacrous area: no details of iris visible: size of pupil barely discernible 3
Opaque cornea: iris not discernible through the opacity 4

F. Area of cornea involved
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

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

> 0.66 - <= 1

Max. score:

1

Reversibility:

fully reversible within: one week

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

>= 0 - <= 0.33

Max. score:

1

Reversibility:

fully reversible within: 24h

Irritation parameter:

other: discharge

Basis:

mean

Time point:

24/48/72 h

Score:

>= 0 - <= 0.66

Max. score:

1

Reversibility:

fully reversible within: 72h

Irritant / corrosive response data:

Treated and control eyes were examined at 1, 24, 48 and 72 hours then 1 week after the application.
One hour after treatment some hyperemic blood vessels (score 1) were observed and the swelling of the conjunctivae was different from normal (score 1) in all animals. The amount of discharge of the conjunctivae was different from normal (score 1) in all animals.
24 hours after treatment some hyperemic blood vessels (score 1) were observed in all animals. The swelling and the amount of discharge of the conjunctivae was different from normal (score 1) in animal No.: 933.
48 hours after treatment some hyperemic blood vessels (score 1) were observed in all animals. The amount of discharge of the conjunctivae was different from normal (score 1) in animal No.: 933.
72 hours after treatment some hyperemic blood vessels (score 1) were observed in two animals (No.: 931, 933). Animal No.: 932 became free of symptoms.
1 week after treatment all animals were free of symptoms.
1 week after the treatment the study was finished.
During the study the control eyes of animals were symptom-free.

Individual scores for ocular irritation 

Study no:                  805-405-2611                 Species:    NZW Rabbit

Dose:                        0.1 g                                Sex:          Male

Test Item:                 p-Nitrobenzoic acid, compound with 2,2’,2’’-nitrilotriethanol (1:1)

 

Time	Animal No.	Score of irritation
		Conjunctivae			Opacity of cornea		Iris	Control eye	Other sign
		R	CH	D	OD	OE	R
1 hour after the treatment	932	1	1	1	0	0	0	0	-
	931	1	1	1	0	0	0	0	-
	933	1	1	1	0	0	0	0	-

 

 

24 hour after the treatment	932	1	0	0	0	0	0	0	-
	931	1	0	0	0	0	0	0	-
	933	1	1	1	0	0	0	0	-

 

 

48 hour after the treatment	932	1	0	0	0	0	0	0	-
	931	1	0	0	0	0	0	0	-
	933	1	0	1	0	0	0	0	-

 

 

72 hour after the treatment	932	0	0	0	0	0	0	0	-
	931	1	0	0	0	0	0	0	-
	933	1	0	0	0	0	0	0	-

 

 

1 week after the treatment	932	0	0	0	0	0	0	0	-
	931	0	0	0	0	0	0	0	-
	933	0	0	0	0	0	0	0	-

Abbreviations:   R    = Redness                                OD =   Opacity degree of density

                           CH = Chemosis                              OE =   Extension of opaque area

D    = Discharge

Mean values of eye irritation scores for EU classificatio

(24, 48 and 72 hours reading)

 

Study no:                  805-405-2611                 Species:    NZW Rabbit

Dose:                        0.1 g                                Sex:          Male

Test Item:                 p-Nitrobenzoic acid, compound with 2,2’,2’’-nitrilotriethanol (1:1)

 

 

Animal Number	Sex	Cornea Opacity	Iris	Conjunctivae
				Redness	Chemosis	Discharge
932	male	0.00	0.00	0.66	0.00	0.00
931	male	0.00	0.00	1.00	0.00	0.00
933	male	0.00	0.00	1.00	0.33	0.66

Body weight data

 

Animal Number	Beginning of study (g)	At the end of study (g)	Body weight gain (g)
932	2560	2898	338
931	2618	2898	280
933	2572	2850	278

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, test item p-Nitrobenzoic acid, compound with 2,2’,2’’-nitrilotriethanol (1:1) applied to the rabbits' eye mucosa caused slight conjunctival irritant effects which were fully reversible within 1 week.

Executive summary:

The acute eye irritation study with test itemp-Nitrobenzoic acid, compound with 2,2’,2’’-nitrilotriethanol (1:1)was performed in New Zealand White rabbits. The irritation effect of the test item was scoring according to the Draize method (Draize, 1959; OECD Guideline 405, 2012).The test item was placed into the conjunctival sac of left eye of each animal. The untreated right eye served as control. 0.1 g of the test item was used for the study in a single dose. The test material was ground to a fine dust. The eyes of the test animals were not washed out.Treated and control eyes were examined at 1, 24, 48 and 72 hours the 1 week after the application.1 hour after the single application of test item into the eye of the rabbits, slight redness, chemosis and discharge occurred in all animals. 72 hours after the treatment slight rednesswas observed in two animals. One animal became free of symptoms. 1 week after the treatment all animals were free of symptoms, so the study was terminated. The animals’ individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

cornea opacity : 0.00, 0.00, 0.00

iris                    : 0.00, 0.00, 0.00

redness             : 0.66, 1.00, 1.00

chemosis          : 0.00, 0.00, 0.33

discharge         : 0.00, 0.00, 0.66

No systemic toxicity was observed on the day of the treatment and during the 1-week observation period. The body weight of animals corresponded to their species and age. Sign of pain and distress as discharge was observed in animals between the treatment day and at first observation of Day 2.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation / corrosion:

In an in vivo skin irritation study in New Zealand White rabbits according to OECD Guideline 404 and EU Method B.4 (Safepharm, 2001), a single 4-hour semi-occluded application (0.5 ml) of the test material was administered to the intact skin of three rabbits. Skin reactions were recorded 1, 24, 48 and 72 hours after administration. The mean value from gradings at 24, 48 and 72 hours after patch removal calculated were the following:

- erythema/eschar: score 0/6=0 (3 out of 3 animals)

- oedema: 0/6=0 (3 out of 3 animals)

No evidence of skin irritation/corrosion was noted. The test material is not corrosive and not irritant to skin according to the regulation 1272/2008/EC.

Eye irritation:

In an acute eye irritation study (Toxicoop, 2017) in New Zealand White rabbits according to the Draize method, 0.1 g the test item was placed into the conjunctival sac of left eye of each animal. The untreated right eye served as control. Treated and control eyes were examined at 1, 24, 48 and 72 hours and 1 week after the application. The animals’ individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

cornea opacity : 0.00, 0.00, 0.00

iris                    : 0.00, 0.00, 0.00

redness             : 0.66, 1.00, 1.00

chemosis          : 0.00, 0.00, 0.33

discharge         : 0.00, 0.00, 0.66

No systemic toxicity was observed on the day of the treatment and during the 1-week observation period.The test material is not irritant to the eyes according to the regulation 1272/2008/EC.

Justification for classification or non-classification

Skin irritation:

According to the in vivo skin irritation study no evidence for skin irritation was noted for p-Nitrobenzoic acid, compound with 2,2',2''-nitrilotriethanol (1:1). The test item did not meet the criteria for classification as irritant or corrosive according to the criteria of the CLP Regulation (EC) No 1272/2008.

Eye irritation:

According to the acute eye irritation study no evidence for eye irritation was noted for p-Nitrobenzoic acid, compound with 2,2',2''-nitrilotriethanol (1:1). The test item did not meet the criteria for classification as irritant or corrosive according to the criteria of the CLP Regulation (EC) No 1272/2008.