[cyclohexane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid, sodium salt

Endpoint summary

Description of key information

In a skin irritation study conducted to OECD test guideline 404 and in compliance with GLP, ALBRITE CIX(N) was not irritating to the eyes (Safepharm, 1992).

[Cyclohexane-1,2 -diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid, sodium salt was found not irritating to the rabbit eye in a test conducted according to OECD guideline 405, and in compliance with GLP (Safepharm, 1982).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

No data

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd., Moston, Sandbach, Cheshire, UK.
- Age at study initiation: 12-16 weeks
- Weight at study initiation: 2.26-2.72 kg
- Housing: individual housing in suspended cages
- Diet: standard laboratory diet, ad libitum
- Water: ad libitum
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22
- Humidity (%): 56-67
- Air changes (per hr): ca. 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

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

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

six

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- % coverage: No data
- Type of wrap if used: The patch was secured in place with a strip of Blenderm surgical adhesive tape, the trunk was then wrapped in an elasticated corset for the duration of the exposure.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test substance was gently swabbed away with cotton wool and distilled water.
- Time after start of exposure: 4 hours.

SCORING SYSTEM: Draize, 1959

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: average of 24, 48 and 72 hour

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

other: average of 24, 48 and 72 hours

Score:

0.6

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

animal #4

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

other: average of 24, 48 and 72 hours

Score:

0.6

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

animal #5

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #6

Time point:

other: average of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Score:

0.2

Reversibility:

fully reversible

Irritant / corrosive response data:

Very slight erythema was noted at four treated skin sites one hour after patch removal and at two treated skin sites at the 24 and 48 hour observations. Very slight oedema was noted at two treated skin sites one hour after patch removal. All treated skin sites appeared normal 72 hours after treatment.

Other effects:

No other effects were reported.

Interpretation of results:

GHS criteria not met

Conclusions:

In a skin irritation study conducted to OECD test guideline 404 and in compliance with GLP, ALBRITE CIX(N) was not irritating to the rabbit skin.

Executive summary:

The primary skin irritation/corrosion of ALBRITE CIX(N) in the rabbit (4-hour semi-occlusive application) was investigated in a study performed according to OECD Testing Guideline No. 404 and under GLP. Six rabbits were exposed to 0.5 ml of the test material, applied onto clipped skin for 4 hours using a semi-occlusive dressing. Observations were made 1, 24, 48 and 72 hours after exposure. No skin irritation was observed after 4 hours exposure. Based on the results of the study, ALBRITE CIX(N) is considered to be not irritating to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3 November 2016 - 22 November 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: in-vitro test

Strain:

other: isolated bovine cornea

Details on test animals or tissues and environmental conditions:

TEST MATERIAL
- Source: slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands)
- Acclimation period: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions. Then, the corneas were dissected and incubated in incubation medium at 32 +/- 1°C in an incubation chamber for 1 hour.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 32 +/- 1°C

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

An amount of 750 µl of either the negative control, positive control or test item was applied directly on the corneas in such a way that the cornea was completely covered.

Duration of treatment / exposure:

The corneas were incubated in a horizontal position for 10 +/- 1 minutes at 32 +/- 1°C.

Duration of post- treatment incubation (in vitro):

120 +/- 10 minutes at 32+/-1°C.

Details on study design:

The aim of this study was to evaluate the ocular irritancy of the test item as measured by its ability to induce opacity and increase permeability in an isolated bovine cornea.

PROTOCOL
- For each treatment group (negative control, positive control and test item), three replicates were used.
- The opacity of each cornea was determined before exposure.
- The corneas were exposed for 10 minutes.
- After the incubation the change in opacity for each individual cornea (including the negative control) was determined.
- Following the final opacity measurement, permeability of the cornea to Na-fluorescein was evaluated.

INTERPRETATION
The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro irritancy score (IVIS)
- A test substance that induces an IVIS > 55 is defined as a corrosive or severe irritant.

Irritation parameter:

in vitro irritation score

Value:

-0.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The negative control showed no irritation (mean IVIS was 0.443). The positive control induced a very severe irritation on the cornea, mean IVIS was 195.5. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. The mean in vitro irritancy score of the test item was -0.8 after 10 minutes of treatment with the test item.

Interpretation of results:

GHS criteria not met

Conclusions:

ALBRITE CIX(N) is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test.

Executive summary:

A GLP compliant in-vitro study was performed according to OECD Guideline 437 in order to evaluate the ocular irritation properties of ALBRITE CIX(N) on an isolated bovine cornea. The possible ocular irritancy of the test item was tested through topical application for approximately 10 minutes. For each treatment group (negative control, positive control and test item), three replicates were used. After removal of the test item and two hours post-incubation, opacity and permeability values were measured. After treatment, the IVIS (in vitro irritancy score) in the controls is within the acceptability criterion. The positive control showed clear irritating effects. ALBRITE CIX(N) showed no effects on the cornea of the bovine eyes. The IVIS score is -0.8 and therefore it can be concluded that ALBRITE CIX(N) is not a severe eye irritant in the BCOP Test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The skin irritation study was conducted according to OECD test guideline 404. New Zealand white rabbits (3 animals/sex) had 0.5ml undiluted test substance applied to the skin for four hours under a semi-occlusive patch. After four hours the remaining test substance was gently swabbed away. Skin irritation reactions were recorded at 1, 24, 48 and 72 hours after patch removal. There were no overt signs of toxicity recorded. Very slight erythema was noted at four treated skin sites one hour after patch removal and at two treated skin sites at the 24 and 48 hour observations. Very slight oedema was noted at two treated skin sites one hour after patch removal. All treated skin sites appeared normal 72 hours after treatment. Overall, [cyclohexane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid, sodium salt was concluded not to be irritating to the skin.

A GLP compliant in-vitro study according to OECD Guideline 437 was performed in order to evaluate the ocular irritation properties of ALBRITE CIX(N) on isolated bovine cornea. Since ALBRITE CIX(N) showed no effects on the cornea of the bovine eyes, it can be concluded that the test substance is not a severe eye irritant or corrosive in the Bovine Corneal Opacity and Permeability test.

Justification for classification or non-classification

Based on the available skin and eye irritation studies, [cyclohexane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid, sodium salt is not classified for skin or eye irritation under Regulation (EC) 1272/2008.