[No public or meaningful name is available]

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 April - 12 June 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

up-and-down procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Harpin-αß protein

IUPAC Name:

Harpin-αß protein

Details on test material:

The test substance EBC 351 is a product formulation containing 11% w/w of the reference substance, 'Cell Free Harpin Extract of Harpinαβ produced by fermentation', and various other non-toxic substances including maltodextrin (listed in Annex IV of Regulation No/ 1907/2006). Therefore, any toxicity seen in the toxicology studies would most likely be caused by the presence of the reference substance rather than the other product co-formulants. Furthermore, ECB 351 contains a higher percentage of Harpin αß and fermentation solids then the reference substance (ca. 11% versus ca. 5%) and therefore the study results would actually over-estimate the toxicity expected if the reference substance itself had been tested. Therefore, results obtained with EBC 351 can be used in support of the reference substance 'Cell Free Harpin Extract of Harpinαβ produced by fermentation'.

Specific details on test material used for the study:

- Test material: EBC 351 (1% Harpin αß)
- Lot No. of test material: DP021112
- Expiration date of the lot: 20 Feb 04
- Appearance: Pale yellow granular solid
- Storage: Refrigerated at 4°C

The test substance EBC 351 is a product formulation containing 11% w/w of the reference substance, 'Cell Free Harpin Extract of Harpinαβ produced by fermentation', and various other non-toxic substances including maltodextrin (listed in Annex IV of Regulation No/ 1907/2006). Therefore, any toxicity seen in the toxicology studies would most likely be caused by the presence of the reference substance rather than the other product co-formulants. Furthermore, ECB 351 contains a higher percentage of Harpin αß and fermentation solids then the reference substance (ca. 11% versus ca. 5%) and therefore the study results would actually over-estimate the toxicity expected if the reference substance itself had been tested. Therefore, results obtained with EBC 351 can be used in support of the reference substance 'Cell Free Harpin Extract of Harpinαβ produced by fermentation'.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Texas Animal Specialties, Humble, Texas
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Approx. 9 weeks old
- Weight at study initiation: 172 - 193 g
- Fasting period before study: 16 hours before dosing
- Housing: Suspended, wire bottom, stainless steel cage; 1 animal per cage
- Diet: PMI Feeds Inc. Formulab #5008 available ad libitum
- Water: Municipal water supply available ad libitum from automatic water system
- Acclimation period: Not specified

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C
- Humidity: 30 - 70%
- Air changes (per hr): 10 - 12 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hour light/ dark cycle

IN-LIFE DATES: From: 06 April 2003 To: 23 April 2003

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 40% w/v

MAXIMUM DOSE VOLUME APPLIED: 12.6 mL/kg dose was administered

Doses:

5050 mg/kg

No. of animals per sex per dose:

3 females per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations for mortality and clinical/ behavioural signs of toxicity: at least three times on the day of dosing (Day 0) and at least once daily thereafter for 14 days
- Frequency of weighing: Individual body weights were recorded just prior to dosing and on days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross necropsy

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 050 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

There was no mortality during the study.

Clinical signs:

other: All animals appeared normal for the duration of the study.

Gross pathology:

The gross necropsy conducted at termination of the study revealed no observable abnormalities.

Other findings:

- Actual temperature: 21 - 26°C
- Actual relative humidity: 30 - 79%

[Deviations of high temperature/ humidity did not affect the study outcome]

Any other information on results incl. tables

Table 1: Body weights, time of death and gross necropsy (Dose level 5050 mg/kg, 12.6 mL/kg)

Animal No.	Date of dosing	Body weights (g)			Time of death*	Gross necropsy findings
		Day 0	Day 7	Final
71-F	7 Apr 03	193	235	253	Day 14	No observable abnormalities
72-F	9 Apr 03	180	217	250	Day 14	No observable abnormalities
73-F	9 Apr 03	172	207	218	Day 14	No observable abnormalities

* Day of dosing is Day 0; Day 14 is terminal sacrifice

 

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral LD50 of the test substance, EBC 351 (1% Harpin ab), is estimated to be greater than 5050 mg/kg in females. Based on this result, the acute oral toxicity of 'Cell Free Harpin Extract of Harpinαβ produced by fermentation' is also expected to be acutely non-toxic and to be above the limit of classification.