Mucorpepsin

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

January 6, 1982 - March 15, 1982

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

- Principle of test: Acute toxicity of Rennilase SP 252 given once orally to rats followed by an observation period of 14 days.
- Short description of test conditions: The rats were deprived of food approximately 18 hours before dosing once orally by gavage. Rennilase SP 252 was dosed in 2.5 g/kg BW, 5 g/kg BW, 10 g/kg BW and 15 g/kg BW (equivalent to 2.4, 4.7, 9.5 and 14.2 g enzyme concentrate dry matter/kg BW). A control group was also included. All rats were carefully observed in the first two hours after dosing and daily in the following 14 days whereupon a gross post mortem examination was performed. All the rats were weighed on the day of dosing (day 1), on day 7 and on the day of termination (day 14).
- Parameters analysed / observed: Clinical observations, post mortem observations and body weight.

GLP compliance:

no

Remarks:

Before GLP was established. Internal quality check was used under this study.

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of mucorpepsin (CAS no.148465-73-0, EC no 642-981-3, enzyme class 3.4.23.23)

Molecular formula:

Not applicable

IUPAC Name:

Active enzyme protein of mucorpepsin (CAS no.148465-73-0, EC no 642-981-3, enzyme class 3.4.23.23)

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.

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.

IUPAC Name:

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

Test material form:

solid: particulate/powder

Details on test material:

- Lot/batch No.: PPR 1285
- Expiration date of the lot/batch: No specific expiry date. At least 10 years or as long as enzyme activity is preserved

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Møllegaard Breeding Centre Ltd., Ejby, Denmark
- Females nulliparous and non-pregnant: Yes
- Weight at study initiation: 47 - 62 g
- Fasting period before study: approximately 18 hours before.
- Housing: Macrolon cages (Type IV)
- Diet: Ad libitum (Brood Stock Feed for Rats and Mice - R3 - Astra Ewos)
- Water: Ad libitum (Adjusted to pH 3 with citric acid)

ENVIRONMENTAL CONDITIONS
Not specified

IN-LIFE DATES: From: 1982-01-28 To: 1982-02-11

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 20 mL/kg bw

MAXIMUM DOSE VOLUME APPLIED: 15 g/kg bw

Doses:

2.4, 4.7, 9.5 and 14.2 g enzyme concentrate dry matter/kg BW

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 1 day, 7 day and 14 day
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: microscopic and microscopic examination.

Statistics:

Body weights were compared between groups and no significant intergroup difference (p<0.05) was seen.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD0

Effect level:

>= 14 200 mg/kg bw

Based on:

other: Enzyme concentrate dry matter

Mortality:

No animals died during the observation period.

Clinical signs:

other: No clinical signs were observed during the study period.

Gross pathology:

Two animals (1 in control and 1 in the group with highest dosage) showed pneumonia like areas, but microscopic findings omitted pneumonia. Perirenal edema was seen in animal from the control group.

Other findings:

Microscopic findings:
Control animal: In the lung, no abnormalities, only a slightly agonal blood aspiration. In the kidney, few degenerative tubuli in the outer zone of cortex predominantly due to pressure from the perirenal edema.
Animal from the highest dosage group: Agonal blood aspiration (no signs of inflammation).

Applicant's summary and conclusion

Interpretation of results:

other: Data adequate, but insufficient for classification

Conclusions:

Rennilase SP 252 did not cause any effects on Wistar rats administered a single oral dose of 14.2 g enzyme concentrate dry matter/kg bw (equivalent to 1356 mg active enzyme protein/kg bw) followed by a 14-day observation period. Therefore, it can be considered as relatively harmless (according to Guidebook: Toxic Substance Control Act, edited by George Dominquez and published by CRC Press, Inc. in 1977).

Executive summary:

The object of this study was to evaluate the acute toxicity in rats of Rennilase SP 252, an enzyme of microbial origin. The test substance, a yellow-brown powder, was dissolved in tap water and given once orally in doses of 2.4, 4.7, 9.5 and 14.2 g enzyme concentrate dry matter/kg BW to groups of 10 male and 10 female Wistar rats. Tap water was given to another group of 10 male and 10 female Wistar rats as a control. The rats were carefully observed in the first two hours after dosing and subsequently daily in the following 14 days. On day 14 they were sacrificed and post mortem examined. The rats were weighed on the day of dosing, on day 7 and on the day of termination. No animals died during the study.

Rennilase SP 252 did not cause any effects on Wistar rats administered a single oral dose of 14.2 g enzyme concentrate dry matter/kg bw (equivalent to 1356 mg active enzyme protein/kg bw) followed by a 14-day observation period. Therefore, it was considered as relatively harmless (according to Guidebook: Toxic Substance Control Act, edited by George Dominquez and published by CRC Press, Inc. in 1977).