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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:
supporting study
Study period:
The study was performed between 14 October 2009 and 17 November 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Study on analogous substance submitted as supporting data only.
Cross-reference
Reason / purpose for cross-reference:
read-across: supporting information

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Date of GLP inspection: 15 September 2009 Date of Signature on GLP certificate: 26 November 2009
Test type:
fixed dose procedure
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
Tetrasodium pyrophosphate
EC Number:
231-767-1
EC Name:
Tetrasodium pyrophosphate
Cas Number:
7722-88-5
Molecular formula:
H4O7P2.4Na
IUPAC Name:
tetrasodium diphosphate
Details on test material:
Sponsor's identification: Tetrasodium pyrophosphate
Description : white powder
Batch number : 449/09
Date received : 18 September 2009
Storage conditions: room temperature in the dark over silica gel

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source:
Harlan Laboratories UK Limited, Bicester, Oxon, UK

- Age at study initiation:
At the start of the study the animals were eight to twelve weeks of age.

- Weight at study initiation:
The bodyweight variation did not exceed ± 20% of the initial/mean bodyweight of any previously dosed animal(s).

- Fasting period before study:
overnight fast immediately before dosing

- Housing:
The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes.

- Diet (e.g. ad libitum):
(2014 Teklad Global Rodent diet supplied by Harlan Laboratories UK Limited, Bicester, Oxon, UK was allowed ad libitum throughout the study.

- Water (e.g. ad libitum):free access to mains drinking water

- Acclimation period:acclimatisation period of at least five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C):
19 to 25°C

- Humidity (%):
30 to 70%

- Air changes (per hr):
The rate of air exchange was at least fifteen changes per hour.

- Photoperiod (hrs dark / hrs light):
lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.


IN-LIFE DATES: From: Day 1 To: Day 14

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle:
For the purpose of the study the test material was freshly prepared, as required, as a suspension in distilled water to give a dose levels of 300mg/kg and 2000mg/kg bodyweight.

- Amount of vehicle:
Not stated

- Justification for choice of vehicle:
Distilled water was the preferred vehicle of the test method.

- Lot/batch no.:
Not stated

- Purity:
Not stated


MAXIMUM DOSE VOLUME APPLIED:
10ml/kg


DOSAGE PREPARATION:
Not applicable

CLASS METHOD:
Not applicable

- Rationale for the selection of the starting dose:
Using available information on the toxicity of the test material, 2000 mg/kg was chosen as the starting dose.
Doses:
Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a suspension in distilled water, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.
Due to mortality and signs of systemic toxicity at a dose level of 2000 mg/kg, a sighting animal was treated at a dose level of 300 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a suspension in distilled water, at a dose level of 300 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.
No. of animals per sex per dose:
1 female at 2000 mg/kg
4 females at 2000 mg/kg
1 female at 300 mg/kg
4 females at 300 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration:
14 days

- Frequency of observations and weighing:
Clinical observations were made ½, 1, 2, and 4 hours after dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.

- Necropsy of survivors performed:
Yes

- Other examinations performed:
Clinical signs, body weight.

Results and discussion

Preliminary study:
A sighting test at a dose level of 2000 mg/kg was performed.
A sighting test at a dose level of 300 mg/kg was performed.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 95% confidence limits not given in study report.
Mortality:
Dose Level - 2000 mg/kg:
One animal was humanely killed two days after dosing. Two animals were found dead three or seven days after dosing.

Dose Level - 300 mg/kg:
There were no deaths.
Clinical signs:
Dose level - 2000mg/kg:
Signs of systemic toxicity noted were hunched posture, ataxia, lethargy, pilo erection, tiptoe gait and dehydration.
Surviving animals appeared normal one or two days after dosing.

Dose level - 300mg/kg
No signs of systemic toxicity were noted during the observation period.
Body weight:
Dose level - 2000mg/kg:
Surviving animals showed expected gains in bodyweight over the observation period

Dose level 300mg/kg:
All animals showed expected gains in bodyweight over the observation period
Gross pathology:
Dose level - 2000mg/kg:
Abnormalities noted at necropsy of the animal that was humanely killed or animals that died during the study were abnormally red lungs, dark liver or patchy pallor of the liver, dark kidneys, gaseous stomach and slight haemorrhage of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study

Dose level - 300mg/kg:
No abnormalities were noted at necropsy
Other findings:
- Organ weights:
Not recorded

- Histopathology:
EXAMPLE: Not recorded

- Potential target organs:
EXAMPLE: Not recorded

- Other observations:
EXAMPLE: None

Any other information on results incl. tables

Table1              Individual Clinical Observations and Mortality Data -2000mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Female

0

0

0

0

H

0

Ä

0

0

0

0

0

0

0

0

0

0

0

2-0

Female

0

HA

HAP

HAPWt

HAPLWt

HAPL
WtDhX*

 

 

 

 

 

 

 

 

 

 

 

 

2-1

Female

0

HAL

HALP

HALP

A

A

H

H

0

0

X

 

 

 

 

 

 

 

2-2

Female

0

HA

HA

HA

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3

Female

0

HAL

HAL

HALP

HA

HA

X

 

 

 

 

 

 

 

 

 

 

 


0= No signs of systemic toxicity

H = Hunched posture

A = Ataxia

L = Lethargy

P = Pilo‑erection

Wt = Tiptoe gait

Dh = Dehydration

X = Animal dead

X* = Animal humanely killed

Ä= Due to a technician error, observation not performed


Table2              Individual Bodyweights and Bodyweight Changes -2000mg/kg

Dose Level

mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight (g)
at Death

Bodyweight Gain (g) During Week

0

7

14

1

2

2000

1-0 Female

182

184

199

 

2

15

2-0 Female

171

-

-

158

-

-

2-1 Female

186

-

-

162

-

-

2-2 Female

165

174

189

 

9

15

2-3 Female

174

-

-

150

-

-



Table3              Individual Necropsy Findings -2000 mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

1-0 Female

Killed Day 14

No abnormalities detected

2-0 Female

Humanely Killed Day 2

Liver: patchy pallor

Stomach: gaseous

Gastric mucosa: haemorrhagic, slight

2-1 Female

Animal found dead Day 7

Lungs: abnormally red

Liver: dark

Kidneys: dark

2-2 Female

Killed Day 14

No abnormalities detected

2-3 Female

Animal found dead Day 3

Lungs: abnormally red

Liver: patchy pallor

Stomach: gaseous

Gastric mucosa: haemorrhagic, slight

 

Table4              Individual Clinical Observations and Mortality Data -300mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300

3-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0= No signs of systemic toxicity


Table5              Individual Bodyweights and Bodyweight Changes-300mg/kg

Dose Level mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Gain (g) During Week

0

7

14

1

2

300

3-0 Female

169

171

182

2

11

4-0 Female

180

188

195

8

7

4-1 Female

177

182

187

5

5

4-2 Female

184

195

203

11

8

4-3 Female

160

166

173

6

7

 


Table6              Individual Necropsy Findings-300mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

300

3-0 Female

Killed Day 14

No abnormalities detected

4-0 Female

Killed Day 14

No abnormalities detected

4-1 Female

Killed Day 14

No abnormalities detected

4-2 Female

Killed Day 14

No abnormalities detected

4-3 Female

Killed Day 14

No abnormalities detected


Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg bodyweight (EU CLP - Category 4).
Executive summary:

Introduction. The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat. The method was designed to meet the requirements of the following:

OECD Guidelines for Testing of Chemicals No 420 “Acute Oral Toxicity - Fixed Dose Method” (2001)

Method B1bisAcute Toxicity (Oral) of CommissionRegulation (EC) No. 440/2008

Method. Following a sighting test at dose levels of 2000 mg/kg and 300 mg/kg, further groups of four fasted females were given a single oral dose of test material, as a suspension in distilled water, at dose levels of 2000 mg/kg and 300 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. Three animals treated at a dose level of 2000 mg/kg were humanely killed or found dead during the study. There were no deaths noted at a dose level of 300 mg/kg.

Clinical Observations. Signs of systemic toxicity noted in animals treated at a dose level of 2000 mg/kg were hunched posture, ataxia, lethargy, pilo-erection, tiptoe gait and dehydration. There were no signs of systemic toxicity noted at a dose level of 300 mg/kg.

Bodyweight. Surviving animals showed expected gains in bodyweight.

Necropsy. Abnormalities noted at necropsy of animals treated at a dose level of 2000 mg/kg that died during the study or was humanely killed were abnormally red lungs, dark liver or patchy pallor of the liver, dark kidneys, gaseous stomach and slight haemorrhage of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Conclusion. The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat was estimated to be in the range of 300 ‑ 2000 mg/kg bodyweight (EU CLP - Category 4).