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Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1 Jun 2009 - 2 Jul 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
GLP compliance:
yes
Test type:
up-and-down procedure

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
-Total Number: Six
Gender: Female
Age Range: 9 to 11 weeks at start of dosing; records of dates of birth for animals used in this study are retained in the Calvert archives.
Body Weight Range: 164 to 212 grams
Animal Source: Harlan
Experimental History: Purpose-bred and experimentally naïve at the outset of the study.
Identification: Ear tag and cage card.

ENVIRONMENTAL CONDITIONS
Housing: Animals were group housed by sex upon receipt and individually housed upon assignment to study in compliance with the National Research Council “Guide for the Care and Use of Laboratory Animals”. Calvert is a USDA registered and fully AAALAC accredited facility. The room in which the animals were kept was documented in the study records. No other species were kept in the same room.
Lighting: 12 hours light/12 hours dark
Room Temperature: 17.8 to 25.6°C
Relative Humidity: 16 to 78%
Food: All animals had access to Harlan Teklad Rodent Diet (certified) as per Calvert SOP. Animals were fasted overnight prior to dose administration. Food was returned to the animals ~30 minutes after dosing. The lot number(s) and specifications of each lot used are archived at Calvert. No contaminants were known to be present in the certified diet at levels that would be expected to interfere with the results of this study. Analysis of the diet was limited to that performed by the manufacturer, records of which are maintained in the Calvert archives.
Water: Tap water was available ad libitum, to each animal via an automatic watering device. The water is routinely analyzed for contaminants as per Calvert SOP’s. No contaminants were known to be present in the water at levels that would be expected to interfere with the results of this study. Results of the water analysis are maintained in the Calvert archives.
Acclimation: Study animals were acclimated to their housing for a minimum of 5 days prior to dosing.

IN-LIFE DATES: From: 1 Jun 2009 To: 2 Jul 2009

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Jeffsol - Propylene Carbonate - NF
Details on oral exposure:

The 175 mg/kg dose level was prepared at 35 mg/ml and dosed at 5 ml/kg (dose volume). The 550, 2000 and 5000 mg/kg levels were dosed as received at 0.47, 1.7 or 4.3 ml/kg, respectively. The 175 mg of test article was brought to a volume of 5 ml with the vehicle. Each preparation was made daily on the day of dosing. The stock bottle was inverted several times prior to dispensing. The 175 mg/kg formulation was described as a clear liquid. The test article stock bottle was purged with nitrogen after use.

It was important to use an 'anhydrous' or low moisture vehicle in order to prevent hydrolysis of MDI to MDA. Therefore no preparations of the test article were made in conjunction with water. All preparations including the test article were made in the Sponsor’s vehicle (Jeffsol - Propylene Carbonate - NF).
Doses:
175, 550, 2000 and 5000 mg/kg
No. of animals per sex per dose:
Dose No. of
Treatment (mg/kg) Sex Rats
Test Item 175 F 1
550 F 1
2000 F 1
5000 F 3
Control animals:
no
Details on study design:
A total of six rats received the test article at dose levels of 175, 550, 2000 or 5000 mg/kg. The test article/dosing suspensions were administered on Day 1 to each rat as a single dose via oral gavage. Animals were fasted overnight prior to dose administration. Each animal received its designated dose based on fasted body weight determined just prior to dosing. The dose volumes were between 0.47and 5 ml/kg and were based on the specific density of 1.17 g/ml as per the MSDS.
The first animal was dosed at an initial dose level of 175 mg/kg. Since this animal survived, the second animal received a higher dose (550 mg/kg) and the third received the test article at 2000 mg/kg. One animal followed by two additional animals were dosed at 5000 mg/kg. The dose for each successive animal was adjusted up or down, depending on the previous result. The test continued based on the fixed time interval outcomes of all the animals up to that point and until one of the stopping criteria was first met. There were three possible stopping criteria.
• 3 consecutive animals survive at the limit dose;
• 5 reversals occur in any 6 consecutive animals test;
• or at least 4 animals have followed the first reversal and the specific likelihood-ratios exceed the critical value.

Mortality checks were made once daily. Clinical observations were recorded prior to dosing and at 30 minutes and 4 hours post-dose, and daily thereafter through Day 15. Body weights were recorded on the day of dosing (Day 1), and on Days 8 and 15. All rats were euthanized by CO2 asphyxiation and necropsied on Day 15.
Statistics:
Dose progression and stopping criteria was calculated using a dedicated software program [Acute Oral Toxicity (Guideline 425) Statistical Program, Version 1.0] provided by the EPA.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Remarks on result:
other: practically nontoxic
Mortality:
Mortality was not observed in any of the animals dosed at 175, 550, 2000 or 5000 mg/kg of the test article.
Clinical signs:
All animals appeared normal throughout the study at 175, 550 and 2000 mg/kg. Clinical signs of piloerection were observed in one of the three animals dosed at 5000 mg/kg at 4 hours post-dose.
Body weight:
No biologically significant effect was seen on body weights of animals on Days 8 and 15.
Gross pathology:
Terminal necropsy revealed no visible lesions in any of the animals at 175, 550 2000 or 5000 mg/kg.

Any other information on results incl. tables

Mortality Summary Report

 

Dose

 

No. of

Observation Period

Total

Treatment

(mg/kg)

Sex

Rats

Day
1

Day
2

Day
3

Day
4

Day
5

Day
6

Day
7

Day
8

Day
9

Day
10

Day
11

Day
12

Day
13

Day
14

Day
15

Mortality

Test Item

175

F

1

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/1

550

F

1

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/1

2000

F

1

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/1

5000

F

3

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/3

Individual Clinical Observation

  

Animal ID#

Observation Period (Day)

Treatment
175 mg/kg

Day 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

30 Min

4 Hr

3251F

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

 

 

Animal ID#

Observation Period (Day)

Treatment
550 mg/kg

Day 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

30 Min

4 Hr

3252F

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

 

 

Animal ID#

Observation Period (Day)

Treatment
2000 mg/kg

Day 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

30 Min

4 Hr

3253F

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

All animals appeared normal prior to dosing   AN = Appears normal   PIL = Piloerection   SS = Soft stools

Animal ID#

Observation Period (Day)

Treatment
5000 mg/kg

Day 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

30 Min

4 Hr

3254F

AN

PIL

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

3255F

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

3256F

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

AN

All animals appeared normal prior to dosing     AN = Appears normal     PIL = Piloerection

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Criteria used for interpretation of results: OECD GHS
Conclusions:
Based on the results of this study, the oral LD50 for the test item in rats was estimated to be greater than 5000 mg/kg.
Executive summary:

The purpose of this study was to assess the toxicity of the test article following a single oral dose to the rat. The results of the study are believed to be of value in predicting the likely toxicity in man by the oral route.

Initially, one female Sprague Dawley rat was dosed at 175 mg/kg. No mortality was observed at 175 mg/kg and dosing continued in another female at 550 mg/kg. Dosing continued in an additional female at 2000 mg/kg as per protocol guidelines and then in one female at 5000 mg/kg. Based on these results two additional females were dosed at 5000 mg/kg. A total of 6 females were dosed. Mortality checks were made once daily. Clinical observations were recorded prior to dosing, as well at 30 minutes, 4 hours post-dose, and daily thereafter through Day 15. Body weights were recorded on the day of dosing (Day 1), and on Days 8 and 15. All rats were euthanized by CO2asphyxiation and necropsied on Day 15.

For doses of 175, 550, 2000 or 5000 mg/kg, no mortality was observed. All animals appeared normal throughout the study at 175, 550 and 2000 mg/kg. Clinical signs of piloerection were observed in one of the three animals dosed at 5000 mg/kg at 4 hours post-dose. No biologically significant effect was seen on body weights on Days 8 and 15. Terminal necropsy revealed no visible lesions in any of the animals at 175, 550, 2000 and 5000 mg/kg.