Amylase, gluco-

Endpoint summary

Description of key information

Glucoamylase did not exert any skin and eye irritation.

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:

Sep. 30 - Oct. 10, 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Glucoamylase is also used in animal feed. Thus, in vivo skin irritation study were required to assess worker safety according to the provisions of Regulation EC 429/2008. Also, the findings from these studies were used for product classification and labelling in the US (OSHA, HSC).

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

April 2002

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: Bred by Charles River, Germany
- Weight at study initiation: between 3.9-4.0 kg
- Age at study initiation: Adults
- Housing: Individually, in animal room with control of temperature (18-24°C) and humidity (40-70%RH)
- Diet (e.g. ad libitum): Standard diet ad libitum
- Air changes (per hr): 10 per hr
- Water (e.g. ad libitum): Acidified tap water ad libitum
- Acclimation period: At least 12 weeks
- Photoperiod (hrs dark / hrs light): 12 hrs/12hrs

IN-LIFE DATES: From: 2008-09-30 To: 2008-10-10

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): undiluted, i.e. 199.63 mg total protein/mL

Duration of treatment / exposure:

4 hours

Observation period:

72 hrs

Number of animals:

3

Details on study design:

The test compound glucoamylase was assessed by semiocclusive application of 0.5 mL of the test material formulation to the closely-clipped dorsa of three New Zealand White rabbits for 4 hrs.
The study was conducted to comply with the following guidelines:
- Organisation for Economic Co-operation and Development (OECD) test guideline 404.

Irritation parameter:

erythema score

Basis:

animal #1

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:

erythema score

Basis:

animal #2

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:

erythema score

Basis:

animal #3

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:

No erythema, eschar formation or oedema was observed at the test sites at any of the three rabbits, neither at termination of exposure or at the examinations 1 hour, 24, 48 or 72 hours after termination of exposure.

Other effects:

No other effects.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material glucoamylase shall be classified as non-irritant according to the Commission Directive 2001/59/EC of 6 August 2001 adapting to the technical progress for the 28th time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances.

Executive summary:

The acute dermal irritant effect of glucoamylase was investigated according to the method recommended in the OECD Guideline No 404, “Acute Dermal Irritation/Corrosion”. The study was conducted in accordance with GLP.

The study was initiated with one male New Zealand White rabbit, which was exposed to 0.5 mL of the undiluted liquid test item applied under semiocclusive conditions to each of three closely-clipped test sites, 2.5 x 2.5 cm, on the back. A fourth similar area of skin served as negative control. After specified periods of application (respectively 3 minutes, 1 hour and 4 hours of exposure), the test item was removed from the test sites and the skin was examined. No erythema, eschar formation or oedema was observed at any of the test sites at any of the examinations. Two additional male animals were then included in the study and treated in the same manner. No erythema, eschar formation or oedema was observed at the test or control sites of any of the animals, neither at termination of exposure or at the examinations 1 hour, 24, 48 or 72 hours after termination of exposure.

The Test Item Mean Score for erythema after 4hrs of exposure to the test item was: 0.0

The Test Item Mean Score for oedema after 4hrs of exposure to the test item was: 0.0

In conclusion, glucoamylase shall be classified as non-irritant.

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:

October 21-31 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Glucoamylase is also used in animal feed. Thus, in vivo skin irritation study were required to assess worker safety according to the provisions of Regulation EC 429/2008. Also, the findings from these studies were used for product classification and labelling in the US (OSHA, HSC).

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

April 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred by Charles River, Germany
- Age at study initiation: Adults
- Weight at study initiation: 3.4-4.5 kg
- Housing: individually in PPO-plast cages (Noryl®; area 4200 cm2) with perforated floor
- Diet (e.g. ad libitum): Standard diet ad libitum
- Water (e.g. ad libitum): Acidified tap water ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24°C
- Humidity (%): 40-70 %
- Air changes (per hr): 10 air changes per hr
- Photoperiod (hrs dark / hrs light): 12 hrs light/ 12 hrs dark cycle

IN-LIFE DATES: From: 2008-10-21 To: 2008-10-31

Vehicle:

other: Tested neat

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): Undiluted test material, i.e. 199.63 mg total protein/mL

Observation period (in vivo):

The reactions were judged 1, 24, 48 and 72 hours after treatment

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No washing

SCORING SYSTEM: According to OECD 405

Irritation parameter:

cornea opacity score

Basis:

animal: 1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: 1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

(Redness of conjunctiva)

Basis:

animal: 1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal: 1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

At the 1 hour reading, discharge was present (score 1) in rabbit # 2. However, this symptom had disappeared at the following observations (24, 48 and 72 hours after treatment). At the examination 1 hour and 24 hours after exposure, discharge was present (score 1) in rabbit #3. However these symptoms had disappeared at the following observations 48 and 72 hours after treatment. No other effects were seen in any of the animals at any of the readings.

Interpretation of results:

GHS criteria not met

Conclusions:

According to the Commission Directive 2001/59/EC of 6 August 2001, 3.2.6.2 Ocular lesions, Glucoamylase should not be classified as eye irritating.

Executive summary:

The study was carried out according to OECD guideline no. 405. Three rabbits were treated as follows: One rabbit was treated first and two more the following day. An amount of 0.1 mL of the undiluted test substance was instilled in the conjunctival cul-de-sac of one of the eyes of the rabbit. After administration, the upper and the lower eye lid were carefully closed and subsequently held together for at least one second before releasing to prevent loss of material. The other eye remained untreated and served as a control. The reactions of the eyes were judged at 1 h, 24 h, 48 h and 72 h after treatment. At the examination 24-hour after treatment, instillation of Fluorescein followed by flushing with 0.9% NaCl was included. UV-light was used to detect possible corneal damage.

The only effects seen were ocular discharge (score 1) in two rabbits at the 1 hour reading, still present in one rabbit at the 24 hr reading. However these symptoms had disappeared at the following observations 48 and 72 hours after treatment. No other effects were seen in any of the animals at any of the readings.

According to the Commission Directive 2001/59/EC of 6 August 2001, 3.2.6.2 Ocular lesions, glucoamylase should not be classified as eye irritating.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

The skin and eye irritation potential of glucoamylase has been tested according to OECD guidelines, and in compliance with GLP. The only reaction seen was transient discharge (score 1) in 2 rabbits in the eye irritation study (totally cleared at 48h), all other readings in both studies were scored 0. The conclusion was that glucoamylase did not exert any skin and eye irritation.

Justification for classification or non-classification

The skin and eye irritation potential of glucoamylase has been tested according to OECD guidelines, and in compliance with GLP.

The conclusion was that glucoamylase did not exert any skin and eye irritation.