Butane-1,2,3,4-tetracarboxylic acid

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

data is from NTRL reports

Data source

Reference

Reference Type:

secondary source

Title:

Eye irritation test in Rabbits (Draize and Kelly Modified)

Author:

NTRL

Year:

1979

Bibliographic source:

OTS – 0280 – 0059 – IN, NTRL, last updated 1979

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Draize and Kelly Modified method

Principles of method if other than guideline:

To assess the ocular irritation potential of Butane-1,2,3,4-tetracarboxylic acid in rabbits according to Draize and Kelly Modified method

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Butane-1,2,3,4-tetracarboxylic acid

EC Number:

216-938-0

EC Name:

Butane-1,2,3,4-tetracarboxylic acid

Cas Number:

1703-58-8

Molecular formula:

C8H10O8

IUPAC Name:

butane-1,2,3,4-tetracarboxylic acid

Test material form:

solid

Details on test material:

- Name of the test chemical: Butane-1,2,3,4-tetracarboxylic acid- IUPAC name: Butane-1,2,3,4-tetracarboxylic acid - Molecular Formula: C8H10O8- Molecular Weight: 234.159 g/mol- Smile Notation: OC(=O)C[C@H]([C@H](CC(=O)O)C(=O)O)C(=O)O- InChI : 1S/C8H10O8/c9-5(10)1-3(7(13)14)4(8(15)16)2-6(11)12/h3-4H,1-2H2,(H,9,10)(H,11,12)(H,13,14)(H,15,16)/t3-,4+- Substance type: organic

Specific details on test material used for the study:

Name of the test chemical: Butane-1,2,3,4-tetracarboxylic acidMolecular Formula: C8H10O8Molecular Weight: 234.159 g/molSmile Notation: OC(=O)C[C@H]([C@H](CC(=O)O)C(=O)O)C(=O)OInChI : 1S/C8H10O8/c9-5(10)1-3(7(13)14)4(8(15)16)2-6(11)12/h3-4H,1-2H2,(H,9,10)(H,11,12)(H,13,14)(H,15,16)/t3-,4+Substance type: organic Physical state: Solid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Sex: Male and femaleSource: No dataAge at study initiation: young adult stageWeight at study initiation: No dataFasting period before study: No dataHousing: metal cages elevated above droppingsDiet (e.g. ad libitum):Purina Rabbit Chow, ad libitumWater (e.g. ad libitum): Tap Water, ad libitumAcclimation period: No dataENVIRONMENTAL CONDITIONSTemperature (°C): No dataHumidity (%): No dataAir changes (per hr): No dataPhotoperiod (hrs dark / hrs light): No dataIN-LIFE DATES: From: To: No data

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

One-tenth mg of undiluted Powdered sample

Duration of treatment / exposure:

single exposure

Observation period (in vivo):

1,24,48, 72 hours and 7,10 and 14 days

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

9 rabbits

Details on study design:

TEST SITE- Area of exposure: conjunctival sacREMOVAL OF TEST SUBSTANCE- Washing (if done): yes, for 3 rabbits- Time after start of exposure: the treated eyes of 3 animals were washed with 120 ml. of lukewarm tap water (approximately at body temperature) at twenty seconds after instillation.Fluorescein test: The treated eyes of the nine animals were further examined by placing one drop of Fluorescein Sodium - 2% on the cornea. After fifteen seconds, the excess fluorescein was flushed out with sufficient isotonic saline.SCORING SYSTEM: The eyes were examined and the grade of ocular reaction was recorded at 1, 24, 48, 72 and 96 hours and 7, 10 and 14 days, using the scale for scoring ocular lesions as outlined by Lehman, A.J. et al., in Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.Table 1: Scale for scoring of Ocular lesions**Cornea Grade (A)Opacity: degree of density (readings should be taken from most dense area)* No ulceration or opacity = 0 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris slightly obscured = 1 Easily discernible translucent area; details of iris slightly obscured = 2 Opalescent area; no details of iris visible; size of pupil barely discernible = 3 Opaque cornea; iris not discernible through the opacity = 4 (B)Area of cornea involvedOne quarter (or less) but not zero = 1Greater than one quarter, but less than half = 2Greater than half, but less than three quarters = 3Greater than three quarters, upto whole area = 4Score equals A*B*5 = Total maximum - 80Iris (A)ValuesNormal = 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 = 2Score Equals A*5 = Total maximum - 10Conjunctivae (A)Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and iris) Vessels Normal = 0 Vessels definitely injected above normal = 1 Diffuse, crimson colour; individual vessels not easily discernible = 2 Diffuse beefy red = 3 (B)Chemosis Swelling (refers to lids and/or nictating membranes) Normal = 0 Any 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)DischargeNo discharge = 0Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) = 1Discharge with moistening of the lids and hairs just adjacent to lids = 3Discharge with moistening of the lids and hairs, and considerable area around the eye = 4Score Equals (A+B+C)*2 = Total maximum – 20The maximum total score is the sum of all scores obtained for the cornea, iris, conjunctivae.Overall Maximum Possible Score = 110** scale for scoring ocular lesions as outlined by Lehman, A.J. et al., in Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.

Results and discussion

In vitro

Other effects / acceptance of results:

no data available

In vivo

Results

Results 1

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

72 h

Score:

85.3

Max. score:

110

Reversibility:

not fully reversible within: 14 days

Remarks on result:

probability of severe irritation

Remarks:

for unwashed eyes

Results 2

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

48 h

Score:

26.7

Max. score:

110

Reversibility:

not fully reversible within: 14 days

Remarks on result:

probability of severe irritation

Remarks:

for washed eyes

Irritant / corrosive response data:

The treated eyes were not washed following instillation of the test material, produced within 24 hours an ocular reaction in the form of a slight to moderate erythema of the bulbar and palpebral conjunctivae, a severe chemosis of the lids, a moderate to marked accumulation of watery-mucoid discharge and a moderate to marked corneal opacity involving one fourth to all of the corneal surface. At 48 hours the eyes reaction had become more pronounced with 5 of 6 test eyes developing iritis. The maximum reaction was observed at 72 hours and only slight improvement was observed through the 14 days. Vascularization was first observed at 96 hours and continued to develop through 14 days. The treated eyes washed at twenty seconds after instillation of the test material, produced within 24 hours an ocular reaction in the form of a slight to moderate erythema of the bulbar and palpebral conjunctivae, a moderate to severe chemosis of the lids, a slight to moderate accumulation of watery-mucoid discharge and a slight to moderate corneal opacity involving one-fourth to one-half of the corneal surface. At 48 hours the reaction of the conjunctivae became more intense where as the corneal reaction in one animal disappeared, was decreased in one other and became more intense in one animal. By 96 hours only slight improvement was noted. At 7 days, two animals showed only slight erythema as a reaction. The other continued to have corneal opacity. At the 14 day observation period, two animals were reaction free while the other exhibited erythema, chemosis and corneal opacity.

Any other information on results incl. tables

Table 2: Scores for ocular lesions at 1, 24, 48, 72, 96 hours and 7, 10 and 14 days of observation

Rabbit no	Structure	1 hour	24 hours	48 hours	72 hours	96 hours	7 days	10 days	14 days
Treated eyes were not washed following instillation of test compound:
1	Cornea	0	45	60	80	80	80	80	80
	Iris	0	0	5	5	5	5*	5*	5*
	Conjunctivae	10	14	20	20	18	14	14	16
2	Cornea	0	15	15	15	15	10	10	10
	Iris	0	0	0	0	0	0	0	0
	Conjunctivae	10	18	18	14	12	6	6	6
3	Cornea	0	30	30	60	60	60	40	40
	Iris	0	0	5	5	5	0	0	0
	Conjunctivae	8	16	18	20	18	12	14	10
4	Cornea	0	45	60	80	80	80	80	80
	Iris	0	0	5	5	5	5*	5*	5*
	Conjunctivae	10	16	20	18	20	16	18	16
5	Cornea	0	60	80	80	80	80	80	80
	Iris	0	0	5	5	5	5*	5*	5*
	Conjunctivae	10	16	20	20	20	16	12	14
6	Cornea	0	30	45	60	60	45	45	30
	Iris	0	0	5	5	5	0	0	0
	Conjunctivae	10	16	20	20	20	16	12	14
	Average	9.3	53.8	71.8	85.3	84.3	75.0	72.0	69.2
	Range
	Cornea	0	15-60	15-80	15-80	15-80	10-80	10-80	10-80
	Iris	0	0	0-5	0-5	0-5	0-5	0-5	0-5
	Conjunctivae	8-10	14-18	18-20	14-20	12-20	6-16	6-18	6-18
Treated eyes were washed 20 seconds after instillation of test compound:
7	Cornea	0	5	0	0	0	0	0	0
	Iris	0	0	0	0	0	0	0	0
	Conjunctivae	12	8	10	8	6	2	0	0
8	Cornea	0	10	5	10	10	0	0	0
	Iris	0	0	0	0	0	0	0	0
	Conjunctivae	10	10	12	8	4	2	2	0
9	Cornea	0	20	30	30	30	20	20	20
	Iris	0	0	5	5	5	0	0	0
	Conjunctivae	8	16	18	14	14	8	4	4
	Average	10.0	23.0	26.7	25.0	23.0	10.7	8.7	8.0
	Range
	Cornea	0	5-20	0-30	0-30	0-30	0-20	0-20	0-20
	Iris	0	0	0-5	0-5	0-5	0	0	0
	Conjunctivae	8-12	8-16	10-18	8-14	4-14	2-8	0-4	0-4

 

*These values are assumed since the density of the corneal opacity is great enough to obscure reading of the iris.

 

 

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Vascularization was first observed at 96 hours and continued to develop through 14 days.The maximum average score of 85.3 was recorded at 72 hours for unwashed eyes.Vascularization appeared in two of the three animals for the washed group at 96 hours and continued through 14 days.A maximum average score of 26.7 was recorded at 48 hours.Since, the effects in the washed and unwashed eyes were not reversible even after 14 days of observation, Butane-1,2,3,4-tetracarboxylic acid can be considered to be an eye irritant capable of producing permanent damage to the eyes.

Executive summary:

The ocular irritation potential of Butane-1,2,3,4-tetracarboxylic acid was assessed in rabbits according to Draize and Kelly Modified method.One-tenth mg of undiluted Powdered sample was placed in one eye of 9 New Zealand White rabbits by gently pulling the lower lid away from the eyeball to form a cup into which the test material was deposited. The lids were then gently held together for one second and the animal was released. The other eye was not treated and served as a control. Following instillation of the test material, the treated eyes of six animals remained unwashed. The treated eyes of the three remaining animals were each washed with 120 ml. of lukewarm tap water (approximately at body temperature) at twenty seconds after instillation of the test material. A one minute wash process was employed for eachanimal.All animals were immobilized in a suitable restrainer rack for one hour subsequent to treatment.The eyes were examined and the grade of ocular reaction was recorded at 1, 24, 48, 72 and 96 hours and 7, 10 and 14 days, using the scale for scoring ocular lesions as outlined by Lehman, A.J. et al., in Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.

After examination of both eyes of each of the nine animals at twenty-four hours, the treated eyes of the first six rabbits were washed with sufficient clean isotonic saline at body temperature to remove any residual test sample. The treated eyes of each of the nine animals were further examined by placing one drop of Fluorescein Sodium - 2% on the cornea. After fifteen seconds, the excess fluorescein was flushed out with sufficient isotonic saline.The treated eyes were not washed following instillation of the test material, produced within 24 hours an ocular reaction in the form of a slight to moderate erythema of the bulbar and palpebral conjunctivae, a severe chemosis of the lids, a moderate to marked accumulation of watery-mucoid discharge and a moderate to marked corneal opacity involving one fourth to all of the corneal surface. At 48 hours the eyes reaction had become more pronounced with 5 of 6 test eyes developing iritis. The maximum reaction was observed at 72 hours and only slight improvement was observed through the 14 days. Vascularization was first observed at 96 hours and continued to develop through 14 days.The treated eyes washed at twenty seconds after instillation of the test material, produced within 24 hours an ocular reaction in the form of a slight to moderate erythema of the bulbar and palpebral conjunctivae, a moderate to severe chemosis of the lids, a slight to moderate accumulation of watery-mucoid discharge and a slight to moderate corneal opacity involving one-fourth to one-half of the corneal surface. At 48 hours the reaction of the conjunctivae became more intense where as the corneal reaction in one animal disappeared, was decreased in one other and became more intense in one animal. By 96 hours only slight improvement was noted. At 7 days, two animals showed only slight erythema as a reaction. The other continued to have corneal opacity. At the 14 day observation period, two animals were reaction free while the other exhibited erythema, chemosis and corneal opacity.The maximum average score of 85.3 was recorded at 72 hours for unwashed eyes.A maximum average score of 26.7 was recorded at 48 hours for the washed eyes.

Since, the effects in the washed and unwashed eyes were not reversible even after 14 days of observation,Butane-1,2,3,4-tetracarboxylic acid can be considered to be an eye irritant capable of producing permanent damage to the eyes