Fructofuranosidase, β-

Endpoint summary

Description of key information

Skin irritation (OECD 404, GLP), rabbit: not irritating

Read-across substance: Endoxylanase (CAS 9025-57-4)

 

Eye irritation (OECD 405, GLP), rabbit: not irritating

Read-across substance: Endoxylanase (CAS 9025-57-4)

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

refer to analogue justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

The read-across approach is detailed in the analogue justification. The target and source substances are considered unlikely to differ in their skin irritation potential. Based on the results of the available dermal irritation/corrosion study conducted with the source substance endoxylanase (CAS 9025-57-4), the test substance was not irritating to skin. The target substance beta-fructofuranosidase (CAS 9001 -57 -4) is not expected to be hazardous to be irritant to skin..

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Conclusions:

The read-across approach is detailed in the analogue justification. The target and source substances are considered unlikely to differ in their skin irritation potential. Based on the results of the available skin irritation/corrosion study conducted with the source substance endoxylanase (CAS 9025-57-4) the test substance did not show irritant properties towards the skin. The target substance β-fructofuranosidase (CAS 9001-57-4) is not expected to be irritant towards the eyes. CLP: not classified

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 - 22 February 2002

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:

adopted 17 July 1992

Deviations:

yes

Remarks:

occlusive dressing used: adhesive tape, 'Johnson-plast' USP; (Johnson and Johnson Ltd. , Mumbai, India) and a crepe bandage was wrapped around the torso of the animal, this coverage

GLP compliance:

yes (incl. QA statement)

Remarks:

Bundesinstitut für gesundheitlichen Verbraucherschutz und Veterinärmedizin, Berlin, Germany

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxicology department Rallis Research Centre, Bangalore, India
- Age at study initiation: 5 - 6 months
- Weight at study initiation: 2.16 - 2.31 kg
- Housing: individually in fibre board cages (size: L 77 x B 54 x H 63 cm) with steam sterilized paddy husk bedding
- Diet: Rabbit feed (Nav Maharashtra Chakan Oil Mills Ltd., Maharashtra, India), ad libitum
- Water: Deep bore-well water, ad libitum
- Acclimation period: 7 - 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 15 Feb 2002 To: 18 Feb 2002

Type of coverage:

other: adhesive tape, 'Johnson-plast' USP; (Johnson and Johnson Ltd. , Mumbai, India) and a crepe bandage was wrapped around the torso of the animal

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

NEGATIVE CONTROL
- bare cotton gauze of similar size without any test item applied

Duration of treatment / exposure:

4 h

Observation period:

1, 24, 48 and 72 h

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: left side of the dorso-lateral thoracic surface, appriximately 6 cm² (2 x 3 cm)
- Type of wrap if used: adhesive tape, 'Johnson-plast' USP; (Johnson and Johnson Ltd. , Mumbai, India) and a crepe bandage was wrapped around the torso of the animal.

REMOVAL OF TEST SUBSTANCE
- Washing: flushed with deionised water
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1, 24, 48 and 72 h

SCORING SYSTEM:
- Method of calculation: Draize scroing system

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

No local reactions (erythema, edema) indicative for skin irritation have been observed 1 h after patch removal in any animal.

Other effects:

- Other adverse local effects: There were no skin reactions.
- Other adverse systemic effects: There were no pre-termial deaths, no effects on body weight and no signs of toxicity observed. Also at necropsy no abnormaility was detected.

Table 1: Test for Irritant Effects on the Skin (Expos.: 4 h) Draize grade

animal number	1 h		24 h		48 h		72 h
	erythema	edema	erythema	edema	erythema	edema	erythema	edema
1	1	0	0	0	0	0	0	0
2	0	0	0	0	0	0	0	0
3	0	0	0	0	0	0	0	0

 

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Conclusions:

A reliable study conducted in accordance with OECD 404 and GLP, determined the test material to be non-irritating to the skin of rabbits. No edema or erythema were observed at any time point. CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

refer to analogue justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Vehicle:

unchanged (no vehicle)

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: reversibility: not applicable

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: reversibility: not applicable

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

Additional time point 1h (for further details please refer to table 1 in the "Any other information on results incl. tables):
Cornea opacity: mean / of 3 animals was 0.
Iris score: mean / of 3 animals was 0.
Conjuctivae score: mean / of 3 animals 0.
Chemosis score: mean / of 3 animals 0.

Other effects:

- Ophthalmoscopic findings: No eye reactions, indicating irritation, were observed.
- Other observations: No deaths were observed and the body weights of all rabbits increased during the observation period. There were no signs of toxicity observed and necropsy did not reveal any abnormalities.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Conclusions:

The read-across approach is detailed in the analogue justification. The target and source substances are considered unlikely to differ in their eye irritation potential. Based on the results of the available eye irritation/corrosion study conducted with the source substance Endoxylanase (CAS 9025-57-4) the test substance did not show irritant properties towards the eyes. The target substance β-fructofuranosidase (CAS 9001-57-4) is not expected to be irritant towards the eyes. CLP: not classified

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 - 24 February 2002

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:

adopted 24 February 1987

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bundesinstitut für gesundheitlichen Verbraucherschutz und Veterinärmedizin, Berlin, Germany

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Toxicology department Rallis Research Centre, Bangalore, India
- Age at study initiation: 5 - 6 months
- Weight at study initiation: 2.09 - 2.29 g
- Housing: individually in fibre board cages (size: L 77 x B 54 x H 63 cm) with steam sterilized paddy husk bedding
- Diet: Rabbit feed (Nav Maharashtra Chakan Oil Mills Ltd., Maharashtra, India), ad libitum
- Water: Deep bore-well water, ad libitum
- Acclimation period: 7 - 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 15 Feb 2002 To: 24 Feb 2002

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48 and 72 h

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: eyes were irrigated with deionised water
- Time after start of exposure: 24 h

SCORING SYSTEM: Draize scoring system

INTERPRETATION OF RESULTS: Modified version of the Kay and Calandra System. The numeric values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae was designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of opacity). For each tissue the score was calculated as follows:
Score for conjunctivae = (A+ B + C) x 2
Score for iris = D x 5
Score for cornea = (E x F) x 5
Using the numerical data obtained a modified version of the system described by Kay J H and Calandra J C. 1962., (J. Soc. Cosmet. Chem., 13:281 -289) was used to classify the ocular irritancy potential of the test item. This is achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation was calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled the classification of the eye irritancy potential of the test item.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: reversibility: not applicable

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: reversibility: not applicable

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

Additional time point 1h (for further details please refer to table 1 in the "Any other information on results incl. tables):
Cornea opacity: mean / of 3 animals was 0.
Iris score: mean / of 3 animals was 0.
Conjuctivae score: mean / of 3 animals 0.
Chemosis score: mean / of 3 animals 0.

Other effects:

- Ophthalmoscopic findings: No eye reactions, indicating irritation, were observed.
- Other observations: No deaths were observed and the body weights of all rabbits increased during the observation period. There were no signs of toxicity observed and necropsy did not reveal any abnormalities.

Table 1: Results of eye irritation study.

Rabbit #	Time [h]	conjunctivae		iris	cornea
		redness	swelling
1	1	0	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0.0	0.0	0.0	0.0
2	1	0	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0.0	0.0	0.0	0.0
3	1	0	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0.0	0.0	0.0	0.0
	Time [h]	conjunctivae		iris	cornea
		redness	swelling
average score	1	0.00	0.00	0.00	0.00
	24	0.00	0.00	0.00	0.00
	48	0.00	0.00	0.00	0.00
	72	0.00	0.00	0.00	0.00
	24+48+72	0.00	0.00	0.00	0.00

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Conclusions:

Under the conditions of the acute eye irritation/corrosion test, the test substance did not show irritant properties towards the eyes. CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for read-across:

There are no experimental data available regarding the skin and eye irritation/corrosion of β-fructofuranosidase (CAS 9001-57-4). It must be noted however that β-fructofuranosidase is produced by a classical strain of Saccharomyces cerevisiae, which has a long history of safe use in food applications globally. Read-across from an appropriate analogue substance endoxylanase (CAS 9025-57-4) is conducted in accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5 in order to fulfil the standard information requirements defined in Regulation (EC) No 1907/2006, Annex VII and VIII, 8.1.

Common functional groups and structural similarities of the source and target substance are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

 

 

Skin irritation in vivo

In the reliable skin irritation study, performed according to OECD TG 404 and in compliance with GLP, three New Zealand White rabbits were exposed to 0.5 mL of enzyme preparation (CAS 9025-57-4) onto clipped skin (6 cm2) for 4 h (Ravi, 2002). Cotton gauze without any dressing was used as control. Patches were secured to the body of the animal by adhesive tape and a crepe elastic bandage.

Skin reactions were evaluated according to the Draize scoring system 1, 24, 48 and 72 h post-application. No erythema or edema formation and no further local or systemic effects were observed in any animal during the study period. In addition, no deaths were observed during the study period, there were no effects on body weight and at necropsy, there were no abnormalities detected. Based on the study results, the test substance is not considered to be irritating to the skin.

 

A further reliable skin irritation study performed according to OECD TG 404 and in compliance with GLP, three New Zealand White rabbits were exposed to 0.5 g of enzyme preparation containing endoxylanase activity (CAS 9025 -57 -4) onto clipped skin (6 cm2) for 4 h under semi-occlusive conditions (Rees, 1995). Before test substance administration, the skin was moistened with 0.2 mL water. Semi-occlusive dressing without any substance served as control. Skin reactions were evaluated according to the Draize scoring system 1, 24, 48 and 72 h post-application. No erythema or edema formation and no further local or systemic effects were observed in any animal during the study period. Based on the study results, the test substance is not considered to be irritating to the skin.

 

Eye irritation in vivo

In the reliable eye irritation study with enzyme preparation containing endoxylanase activity (CAS 9025-57-4) was performed according to OECD TG 405 and in compliance with GLP (Ravi, 2002). In this study, 0.1 mL of the test substance was instilled in the eyes of three albino rabbits. The untreated eye served as control. After an exposure period of 24 hours, the treated eye was rinsed with deionised water. The eyes were observed and reactions were evaluated according to the Draize scoring system, 1, 24, 48, and 72 h after application. No effects on corneal opacity, conjunctival redness, chemosis and iris were noted at any time point in any animal. No deaths, no local or systemic effects and no necropsy findings were observed in any animal. Based on the obtained results, the test substance is not considered eye irritating.

 

A further reliable eye irritation study with enzyme preparation containing endoxylanase activity (CAS 9025 -57 -4) was performed according to OECD TG 405 and in compliance with GLP (Rees, 1995). In this study, 0.1 g of the test substance was instilled in the eyes of three albino rabbits. The untreated eye served as control. The eyes were observed and reactions were evaluated according to the Draize scoring, 1, 24, 48, 72 h and 8 and 15 days after application. No effects on corneal opacity and chemosis were noted at any time point in any animal. Iritis grade 1 was observed in one animal at the 1 h reading time point, but not at any later time point. Very slight conjunctivitis (redness) was evident in the treated eye of each animal 1 h after instillation. The conjunctival redness resolved by the 48 h observation in one animal and by Day 8 in the remaining rabbits. Based on the obtained results, the test substance is not considered to cause eye irritation requiring classification according to Regulation(EC) No. 1272/2008.

 

Conclusion on skin and eye irritation properties

The read-across approach is detailed in the analogue justification. The target and source substances are considered unlikely to differ in their skin or eye irritation potential. Based on the results of the available skin and eye irritation/corrosion studies conducted with the source substance endoxylanase (CAS 9025-57-4), the test substance did not show irritant properties towards the skin or eyes. Therefore the target substance β-fructofuranosidase (CAS 9001-57-4) is also not expected to be a skin or eye irritant. CLP: not classified

 

Justification for classification or non-classification

Based on the available data, the registration substance is considered not to meet the classification criteria according to Regulation (EC) No. 1272/2008 (CLP).