Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20-03-1982 to 29-03-1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1982
Report Date:
1982

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Toxic Substances Control Act, George Dominquez, 1977
Deviations:
no
GLP compliance:
no
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
powder
Details on test material:
- Lot/batch No.: PPZ1281
- Expiration date of the lot/batch: April 1993

Test animals

Species:
mouse
Strain:
NMRI
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Bred by Novozymes A/S (previously known as Novo Industri A/S)
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: Not stated
- Weight at study initiation: 18-22 g
- Fasting period before study: 18 hour fasting prior to dosing
- Housing: Barriered rodent facility with control of temperature, humidity and lighting. The cages were solid bottomed polycarbonate cages with a stainless steel mesh lid. Softwood bark-free fibre were provided as bedding. Aspen chew block and plastic shelter for environmental enrichment.
- Diet: standard diet (Brood Stock Feed for Rats and Mice - R3 - Astra Ewos) ad libitum
- Water: tap water (adjusted to approximately pH 3.0 with citric acid) ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23°C
- Humidity (%): 45-70%

IN-LIFE DATES: From: 1982-01-20 To: 1982-02-04

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Undiluted test material for the top dose.
Doses:
Doses were 1000, 2000, 5000, 15000 mL/kg bodyweight.
No. of animals per sex per dose:
Three groups of 5 male and 5 female and 2 groups of 2 male and 2 female NMRI mice.
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 15 days.
- Frequency of observations and weighing: Thirty minutes after dosing and daily in the following 9 or 15 days the mice were observed for any signs of reaction. The body weight was recorded just before dosing day 1 (all groups) and day 8 (group 1, 4 and 5), but due to a mistake no body weights were recorded day 15 before sacrifice.
- Necropsy of survivors performed: A macroscopic post mortem examina.tion was perforrned on all mice.
Statistics:
Body weight and body weight gain were subjected to statistical anal ysis using Student's t-test comparing group 1 (control) with group 4 and 5.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD0
Effect level:
15 000 mg/kg bw
Based on:
other: Total Organic Solids (TOS)
Mortality:
No deaths or clinical symptoms were seen at anytime during the observation period.
Clinical signs:
No clinical symptoms were seen at anytime during the observation period.
Body weight:
A decrease in body weight and body weight gain was seen in males given the highest dose, 15000 mg/kg, as the only intergroup difference.
Gross pathology:
No dose related macroscopic findings were seen in any of the mice.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
No deaths clinical symptoms or macroscopic findings were seen in any of the mice, but a decrease in body weight gain was seen in the highest dosed males.
As no deaths occured the LD50 value was estimated to be over 15000 mg/kg for both males and females and Novozym 230 could be classified "relatively harmless" according to Guidebook: Toxic Substances Control Act, George Dorninquez, 1977.
Executive summary:

The objective of this study was to evaluate the acute oral toxicity in mice of the enzyme Novozym 230 (batch PPZ1281). The test substance was dissolved in tap water and given once orally in doses of 0, 1000 , 2000, 5000 and 15000 mg/kg body weight to groups of 2 or 5 male and 2 or 5 female NMRI mice. Thirty minutes after dosing and daily in the following 9 or 15 days, the mice were observed for any signs of reaction. The mice were weighed at weekly intervals. They were sacrificed at the end of the observation period and a macroscopic post mortem examination was performed on all the mice.

No deaths clinical symptoms or macroscopic findings were seen in any of the mice, but a decrease in body weight gain was seen in the highest dosed males.

As no deaths occured the LD50 value was estimated to be over 15000 mg/kg for both males and females and Novozym 230 could be classified "relatively harmless" according to Guidebook: Toxic Substances Control Act, George Dorninquez, 1977.