Reaction mass of trisodium 2-(hydroxyphosphinato)succinate and pentasodium 1-(hydroxyphosphinato)butane-1,2,3,4-tetracarboxylate

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From: 7 September 1992 To: 9 October 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Guinea pig maximisation test was performed in 1993 before the REACH Regulation (EC.1907/2006)

Specific details on test material used for the study:

Purity of the test material is based on the phosphonate composition, with no direct measure of % sodium. ITC 288 is not the "Reaction mass of tetrasodium-phosphonoethane-1,2-dicarboxylate and hexasodium-phosphonobutane-1,2,3,4--tetracarboxylate" (EC 410-800-5) rather the substance "Reaction mass of trisodium-phosphonoethane-1,2-dicarboxylate and pentasodium-phosphonobutane-1,2,3,4--tetracarboxylate" (EC 701-079-0) as demonstrated by the manufacturing specs reported in Section 1.2, legal entity composition.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: David Hall Limited, Burton-on-Trent, Staffordshire, U.K.
- Age at study initiation: 8-12 weeks old
- Weight at study initiation: 300-366 g
- Housing: up to 3 per cage
- Diet : ad libitum
- Water : ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 51-75
- Air changes (per hr): 15
- Photoperiod : 12 hrs dark / 12 hrs light


IN-LIFE DATES: From: 7 September 1992 To: 9 October 1992

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

Intradermal induction: 1 % (w/v) in distilled water
Topical induction: 75 % (w/v) in distilled water

Route:

epicutaneous, semiocclusive

Vehicle:

water

Concentration / amount:

Topical challenge: 50 % and 25 % (w/w) in distilled water

No. of animals per dose:

20 (test)
10 (control)

Details on study design:

RANGE FINDING TESTS:
- For intradermal induction: 1% was selected for the main study considering as the highest concentration that did not cause local necrosis, ulceration or systemic toxicity
- For Topical application: 75% was selected for the main study considering as the highest concentration producing only mild to moderate dermal irritation after a 48-hour occlusive exposure
- For Topical Challenge: 25% and 75% were selected for the main study considering as the highest non-irritant concentration.
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 0 and Day 7
- Site: shoulder region
- Concentrations: 1 % (intradermal), 75 % (epidermal)

Intradermal injection/ day 0
3 pairs of intradermal injections (0.1 ml on each side of the mid-line) were made in an area approximately 40 mm x 60 mm on the shoulder region.
Test animals:
1) Freund's Complete Adjuvant plus distilled water in the ratio 1:1
2) a 1 % (w/v) dilution of test material in distilled water
3) a 1 % (w/v) dilution of test material in a 1:1 preparation of Freund's Complete Adjuvant plus distilled water

Control animals:
1) Freund's Complete Adjuvant plus distilled water in the ratio 1:1
2) distilled water
3) Freund's Complete Adjuvant plus distilled water in the ratio 1:1

Epidermal applications/day 7
One week later (Day 7), the same area on the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the test material formulation (75 % w/w in distilled water). The test material formulation (0.2-0.3 ml) was applied on filter paper (WHATMAN No.4: approximate size 40 mm x 20 mm) which was held in place by a strip of surgical adhesive tape (BLENDERM: approximate size 60 mm x 25 mm) and covered with an overlapping lenght of aluminium foil. The patch and foil were further secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 250 mm x 35 mm) wound in a double layer around the torso of each animal. This occlusive dressing was kept in place for 48 hours.

Erythematous reactions were quantified one and twenty-four hours following removal of the patches.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 21
- Exposure period: 24 hours
- Test groups and Control group: a quantity of 0.1-0.2 ml of the test material formulation (50 % w/w in distilled water) was a^pplied to the shorn right flank of each animal on a square of filter paper which was held in place by a strip of surgical adhesive tape. To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 25% (w/w) in distilled water was also similarly applied to a separate skin site on the right shorn flank. The vehicle alone was similarly applied to the left shorn flank.
- Site: right flank (test material), left flank (vehicle)
- Concentrations: 50 and 25 %
- Evaluation (hr after challenge): 24, 48 and 72 hours

Challenge controls:

no data

Positive control substance(s):

not specified

Statistics:

None

Positive control results:

no data

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

50 %

No. with + reactions:

10

Total no. in group:

19

Clinical observations:

no data

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 10.0. Total no. in groups: 19.0. Clinical observations: no data.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

50 %

No. with + reactions:

7

Total no. in group:

19

Clinical observations:

no data

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 7.0. Total no. in groups: 19.0. Clinical observations: no data.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

25 %

No. with + reactions:

1

Total no. in group:

19

Clinical observations:

no data

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25 %. No with. + reactions: 1.0. Total no. in groups: 19.0. Clinical observations: no data.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25 %

No. with + reactions:

2

Total no. in group:

19

Clinical observations:

no data

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 %. No with. + reactions: 2.0. Total no. in groups: 19.0. Clinical observations: no data.

Table 7.4.1/1: Reaction at challenge at 24 h and 48 h in negative control group and test group (25%)

 

Animal number	Reaction at challenge
	24 h	48 h		72 h
Control group
21	0	0	0
22	0	0	0
23	0	0	0
24	0	0	0
25	0	0	0
26	0	0	0
27	0	0	0
28	0	0	0
29	0	0	0
30	0	0	0
Test group (25%)
1	0	0	0
2	0	0	0
3	0	0	0
4	0	0	0
5	0	0	0
6	0	0	0
7	0	0	0
8	0	0	0
9	0	0	0
10	0	0	0
11	0	0	0
12	0	0	0
13	0	0	0
14	0	0	0
15	0	0	0
16	0	0	0
17	1	1	0D
18	0	1	0
19	-	-	-
20	0	0	0

 

- = no data; animal found dead day 14

D = Desquamation

R = extended reaction

Table 7.4.1/2: Reaction at challenge at 24 h and 48 h in negative control group and test group (50%)

 

Animal number	Reaction at challenge
	24 h	48 h		72 h
Control group
21	0	0	0
22	0	0	0
23	0	0	0
24	0	0	0
25	0	0	0
26	0	0	0
27	0	0	0
28	0	0	0
29	0	0	0
30	0	0	0
Test group (50%)
1	1	1	0
2	1	1	0
3	1	1	0
4	1	1	0
5	0	0	0
6	1	1	0
7	0	0	0
8	1R	1R	0 D
9	1	0	0
10	1	1	0
11	1	0	0
12	0	0	0
13	0	0	0 D
14	0	0	0
15	1	0	0
16	0	0	0
17	0	1	0
18	0	1	0
19	-	-	-
20	0	0	0

 

- = no data; animal found dead day 14

D = Desquamation

R = extended reaction

Interpretation of results:

other: lassified in category 1B, H317 ( May cause an allergic skin reaction) according to the CLP regulation (1272/2008)

Conclusions:

Under the conditions of this test, ITC 288 produced a 58 % (11/19) sensitisation rate and was classified in category 1B, H317 ( May cause an allergic skin reaction) according to the CLP regulation (1272/2008) and as a sensitiser to guinea pig skin (Xi, R43) according to the Directive 67/548/EEC.

Executive summary:

In a dermal sensitisation study (Tuffnell PP, 1992), female albino Dunkin-Hartley guinea pigs (20 tested, 10 control) were used to test the skin contact sensitisation potential of ITC 288. The method of Guinea pig maximisation test was used. The concentrations of the test material were 1% for the intradermal induction, 75 % for the topical induction and 50 and 25 % for the topical challenge. One test animal was found dead on day 14. The cause of death was not determined but the absence of this animal was considered not to affect the purpose or integrity of the study. Scattered mild redness and moderate and diffuse redness were elicited by the test material after topical induction. Other adverse reactions noted were bleeding, dried blood and small superficial scattered scabs. After topical challenge (50 % of test material), positive skin responses were noted at the test sites of ten test animals at the 24 -hour observation and persisted in seven test animals at the 48 -hour observation. The reaction extended beyond the treatment site of one test animal at the 24 and 48 -hour observations. Desquamation was noted at the test sites of two test animals at the 72 -hour observation; according to Draize's scale and Magnusson and Kligman's scale this local effect should not be considered as an positive reaction. No adverse reactions were noted at the test sites of control animals at the 24, 48 and 72 -hour observations. After topical challenge (25 % of test material), a positive skin response was noted at the test sites of one test animal at the 24 and 48 -hour observations. A positive response developed at the test site of one more test animal at the 48 -hour observation. Desquamation was noted at the test site of one test animal at the 72 -hour observation. No adverse reactions were noted at the test sites of control animals at the 24, 48 and 72 -hour observations. It should be noted that desquamation observed for two animals at 72 -hour post exposure were discounted for the calculation of the sensitisation index. Initially the study report mentioned 12 animals with positive reactions, and after re-evaluation of the data only 11 showed clear skin sensitisation effect; the latest animal exhibited only desquamation at 72 -hours post exposure without any previous local effects.

Under the conditions of this test, ITC 288 produced a 58 % (11/19) sensitisation rate and was classified in category 1B, H317 ( May cause an allergic skin reaction) according to the CLP regulation (1272/2008) and as a sensitiser to guinea pig skin (Xi, R43) according to the Directive 67/548/EEC.

This skin sensitisation study is classified as acceptable. It satisfies the guideline requirement for a skin sensitisation in the guinea pig.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

One study was available for this endpoint and was considered as the key study (Kr: 1). In a dermal sensitization study (Tuffnell PP, 1992), female albino Dunkin-Hartley guinea pigs (20 tested + 10 control) were used to test the skin contact sensitisation potential of ITC 288/S using the method of guinea pigs maximisation test (OECD 406, method B6 in Commission Directive 84/449/EEC) and in compliance with GLP. In this study, ITC 288/S induced evidence of skin sensitisation in 58 % of the animals at challenge readings (score 1 at 24h and 48 h) following topical application at 50%. Based on this result, ITC 288/S should be considered as a sensitizer by skin contact.

Migrated from Short description of key information:
Positive in guinea pigs maximisation test (OECD 406, Kr: 1), and therefore classified as skin sensitizer.

Respiratory sensitisation

Endpoint conclusion

Additional information:

Migrated from Short description of key information:
No data was available

Justification for classification or non-classification

Skin sensitisation:

ITC 288/S produced a 58 % (11/19) sensitisation rate (OECD 406, Kr:1). Therefore ITC 288/S was classified in category 1B, H317 ( May cause an allergic skin reaction) according to the CLP regulation (1272/2008) and as a sensitiser to guinea pig skin (Xi, R43) according to the Directive 67/548/EEC.

Respiratory sensitisation:

Classification not possible due to lack of data.