Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was performed between 26 June 2012 and 2 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Date of GLP inspection: 19-21 July 2011 Date of Signature on GLP certificate: 31 August 2011
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Sponsor's identification: trisodium hydrogen diphosphate
Description: white powder
Batch number: 147/12
Purity: 95%
Date received: 07 March 2012
Expiry date: 01 February 2014
Storage conditions: room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals 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: 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 300 mg/kg and 2000 mg/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 applicable

- Purity: Not applicable


MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg


DOSAGE PREPARATION: Not applicable

CLASS METHOD: Not applicable

- Rationale for the selection of the starting dose: In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
Doses:
Dosing regimen:
A single animal was treated with the starting dose of 300 mg/kg bw. In this absence of toxicity on this animal an additional animal was treated with 2000 mg/kg bw. As this animal exhibited no toxicity an additional 4 animals were treated with 2000 mg/kg be of the test material (a total of 5 animals were treated with 2000 mg/kg bw). Due to mortality and signs of systemic toxicity in the animals tested at 2000 mg/kg bw an additional group of 4 animals were treated with 300 mg/kg bw (a total of 5 animals were treated with 300 mg/kg bw).
No. of animals per sex per dose:
5 females at 2000 mg/kg
5 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 300 mg/kg was performed and a sighting test at a dose level of 2000 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: Two animals were found dead one or two 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, pilo-erection, ptosis, lethargy and ataxia. Surviving animals appeared normal one, five or six 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 except for one animal which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.

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 animals that died during the study were dark liver, dark spleen, dark kidneys and haemorrhagic gastric mucosa. A raised limiting ridge of the stomach and pale gastric mucosa were noted at necropsy of one animal that was killed at the end of the study. No abnormalities were noted at necropsy of the remaining 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: Not recorded

- Potential target organs: Not recorded

- Other observations: None

Any other information on results incl. tables

Table 1. 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

2-0

Female

0

0

0

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Female

0

0

0

0

0

0

 0

 0

0

0

0

0

0

0

0

0

3-1

Female

0

0

0

0

HP

H

H

HP

H

0

 0

 0

 0

 0

 0

 0

 0

 0

3-2

Female

0

0

0

HAPPtL

X

 

 

 

 

 

 

 

 

 

 

 

3-3

Female

0

0

0

HLP

HAP

X

 

 

 

 

 

 

 

 

 

 

 

 

0 = No signs of systemic toxicity

H = Hunched posture

P = Pilo-erection

Pt = Ptosis

L = Lethargy

A = Ataxia

X = Animal dead 

Table 2. 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

2-0

Female

162

179

187

 

17

8

3-0

Female

186

202

217

16

15

3-1

Female

180

174

213

- 6

39

3-2

Female

165

-

-

160

-

-

3-3

Female

166

-

-

153

-

-

 



Table 3.  Individual Necropsy Findings -2000 mg/kg

 

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

2-0

Female

Killed Day 14

Stomach: raised limiting ridge

Gastric mucosa: pale

3-0

Female

Killed Day 14

No abnormalities detected

3-1

Female

Killed Day 14

No abnormalities detected

3-2

Female

Animal found dead Day 1

Liver: dark

Spleen: dark

Kidneys: dark

Gastric mucosa: haemorrhagic

3-3

Female

Animal found dead Day 2

Liver: dark

Spleen: dark

Kidneys: dark

Gastric mucosa: haemorrhagic

 

 

 

 

 

Table 4. 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

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

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



Table 5. 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

2-0

Female

162

187

202

25

15

3-0

Female

182

199

216

17

17

3-1

Female

188

197

220

9

23

3-2

Female

183

199

214

16

15

3-3

Female

189

214

230

25

16

 



Table 6. Individual Necropsy Findings - 300mg/kg

 

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

300

2-0

Female

Killed Day 14

No abnormalities detected

3-0

Female

Killed Day 14

No abnormalities detected

3-1

Female

Killed Day 14

No abnormalities detected

3-2

Female

Killed Day 14

No abnormalities detected

3-3

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).

This study is considered to be reliable and acceptable for use as a key study in accordance with Regulation (EC) No. 1907/2006 (REACH) and for the purposes of classification and labelling in accordance with Regulation (EC) No. 1272/2008 (EU CLP).
Executive summary:

.