[No public or meaningful name is available]

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

June 29 to August 31, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Read-across to alpha amylase is applied. Data on alpha amylase are considered also to be valid for beta-mannosidase, because both enzymes belong to the same enzyme sub-subclass IUB 3.2.1 and are considered to have a similar toxicological profile also with regard to acute toxicity.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

Dec. 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of alpha-amylase (EC no. 232-565-6, CAS no. 9000-90-2, EC name: Amylase, alpha-, Enzyme Class No.: 3.2.1.1)

Molecular formula:

Not available

IUPAC Name:

Active enzyme protein of alpha-amylase (EC no. 232-565-6, CAS no. 9000-90-2, EC name: Amylase, alpha-, Enzyme Class No.: 3.2.1.1)

Constituent 2

Reference substance name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available

IUPAC Name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 3

Reference substance name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 4

Reference substance name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process

Constituent 5

Reference substance name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Test material form:

liquid

Details on test material:

- Lot/batch No.: PPY 31071
- Expiration date of the lot/batch: 1 June 2020

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River, Germany
- Fasting period before dosing: Overnight
- Housing: A maximum of 6 animals per sex per cage, transparent macrolon cages
- Weight at time of dosing: between 168-174 g
- Housing: In animal room with control of temperature and humidity
- Diet: Standard diet ad libitum
- Water: Tap water ad libitum
- Acclimatization period: 5 days
- Temperature (°C): 20-24°C
- Humidity : 45-70 %

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg

Doses:

Undiluted test material 20 mL/kg body weight, equivalent to 2562 mg enzyme concentrate dry matter/kg body weight or 859 mg aep/kg body weight (limit testing).

No. of animals per sex per dose:

6 (only females)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations for clinical signs of effect: within 1 hour and within 4 hours after dosing and at least once daily throughout the observation period. Weighing: just prior to dosing on day 0 and on days 3, 7 and 14
- Necropsy of survivors performed: yes

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD0

Effect level:

2 562 mg/kg bw

Based on:

other: Enzyme concentrate dry matter

Effect levels 2

Sex:

female

Dose descriptor:

other: Limit test - no effects were seen

Effect level:

> 20 mL/kg bw

Based on:

test mat.

Mortality:

No mortality.

Clinical signs:

other: No clinical signs.

Gross pathology:

No abnormalities.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

No signs of toxicity were observed among the rats treated with a single oral dose of 2562 mg enzyme concentrate dry matter/kg bodyweight (equivalent to 859 mg aep/kg bodyweight), which was the highest possible dose at dose volume 20 mL/kg, using the undiluted test item.

Executive summary:

The objective of this study was to assess the acute toxicity of alpha-amylase when administered as a single oral dose to rats followed by an observation period of 14 days. The purpose of the study was to satisfy regulatory demands because the enzyme is used for production of food in EU.

The study was conducted in accordance with the OECD Guideline No 423, “Acute Oral Toxicity – Acute Toxic Class method”. The design of the limit test was used. The test item was supplied as a brown liquid ready to use. The dose volume administered was 20 mL/kg body weight corresponding to a dose of 2562 mg enzyme concentrate dry matter/kg body weight (equivalent to 859 mg aep/kg body weight). This was the highest possible dose level at dose volume 20 mL/kg body weight, using the undiluted test item.

No mortality or clinical signs were observed after treatment and the overall body weight gain during the study was considered to be normal. The necropsy revealed no abnormalities.

In conclusion, no signs of toxicity were observed among the rats treated with a single oral dose of 2562 mg enzyme concentrate dry matter/kg body weight, which was the highest possible dose at dose volume 20 mL/kg, using the undiluted test item.