Registration Dossier

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Administrative data

Description of key information

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
29th June 1973
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
The diluted compound was fed by the stomach to male and female rats, after determining the minimal lethal dose, four seperate dose groups (mg/kg) were each administered as a 25% aqueous solution of the test material. Mortalities within 7 days were used to determine the LD50 value for the test material. Observations were also made for toxic signs and the viscera of the test animals were examined macroscopically.
GLP compliance:
no
Remarks:
predates GLP
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
no data
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 25% aqueous solution were prepared
- Amount of vehicle (if gavage): no data

MAXIMUM DOSE VOLUME APPLIED: 3980 mg/kg bw, 220 gm of test material

- Rationale for the selection of the starting dose: The minimal lethal dose was determined and used to derive a range of doses over which the LD50 would be determined
Doses:
2000 mg/kg; 2510 mg/kg; 3160 mg/kg; 3980 mg/kg
No. of animals per sex per dose:
5 animals per dose.
2000 mg/kg - 3 female, 2 male
2510 mg/kg - 2 female, 3 male
3160 mg/kg - 3 female, 2 male
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 7 days
- Necropsy of survivors performed: yes
- Other examinations performed: Clinical observations, the viscera were examined macroscopically and autopsy.
Statistics:
The LD50 was calculated according to the method of E. J. De Beer.
Sex:
male/female
Dose descriptor:
approximate LD50
Effect level:
2 980 mg/kg bw
95% CL:
> 2 740 - < 3 250
Mortality:
yes, see the table in the results section for further details
Clinical signs:
Toxic signs included reduced appetite and activity (one to two days in survivors), increasing weakness and collapse.
Body weight:
no data
Gross pathology:
At autopsy there was hemorrhagic areas of the lungs, slight liver discolouration, and acute gastrointestinal inflammation. The viscera appeared normal by macroscopic examination

The single oral LD50 of tetrapotassium pyrophosphate

sample fed as a 25% aqueous solution

Animal No. – Sex

Weight gm

Dose mg/kg

Fate

1 – female

220

2000

survived

2 – male

205

2000

survived

3 – female

210

2000

survived

4 – male

215

2000

survived

5 – female

225

2000

survived

 

 

 

 

6 – male

235

2510

survived

7 – female

240

2510

died

8 – male

250

2510

survived

9 – female

240

2510

survived

10 – male

230

2510

survived

 

 

 

 

11 – female

260

3160

died

12 – male

215

3160

survived

13 – female

225

3160

died

14 – male

220

3160

died

15 – female

210

3160

survived

 

 

 

 

16 – male

210

3980

died

17 – female

220

3980

died

18 – male

215

3980

died

19 – female

220

3980

died

20 – male

210

3980

died

The compound was classed as slightly toxic by oral ingestion in male and female rats.

Survival time was one to two days.

Interpretation of results:
GHS criteria not met
Conclusions:
The single LD50 for male and female rats was placed at 2980 mg/kg bw, with upper and lower limits of 2740 and 3250 mg/kg bw respectively. Based on this result the compound was described as slightly toxic. In accordance with Regulation (EC) No. 1272/2008 tetrapotassium pyrophosphate is not considered to be toxic via the oral route.
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.
Reason / purpose for cross-reference:
read-across: supporting information
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
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
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.
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.
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

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


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).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
No data.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Guideline study with acceptable restrictions. Substance identification is not fully defined (trade name only) and although it is known to be tetrapotassium pyrophosphate the data should not be used as a key study however the data is acceptable for use as part of a weight of evidence approach (as detailed in Annex XI, Section 1.2 of Regulation (EC) No. 1907/2006).
Qualifier:
according to guideline
Guideline:
other: Code of Federal Regulations, section 1500.
Deviations:
not specified
Principles of method if other than guideline:
Twenty Wistar-derived rats were fasted for 24 h prior to administration of the test substance by oral gavage, doses ranged from 1.74 to 3.48 gm/kg bw . Each animal received the desired dosage in a single administration. The animals were observed for signs of toxicity and survival at 1, 3, 6, 24, 48 and 72 h and daily thereafter for a total of 14 days. Body weights were recorded on day 0, 7 and 14 of the study and all animals were autopsied and observed for gross pathological organ changes.
GLP compliance:
no
Remarks:
study predates GLP
Test type:
other: No data.
Limit test:
no
Species:
rat
Strain:
other: Wistar derived albinos
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Zartman Farms, Douglasville, Pa.
- Age at study initiation: no data
- Weight at study initiation: 200-300 grams
- Fasting period before study: 24 hours.
- Housing: Animals housed individually
- Diet (e.g. ad libitum): ad libitum. The animals were fed a standard laboratory diet of 'Purina Rat Chow'.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
no data

IN-LIFE DATES: From: no data
Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
No data.
Doses:
There were 5 dosing groups:
1.74, 2.17, 2.61, 3.04, 3.48 gm/kg bw
No. of animals per sex per dose:
Two animals per sex per dose.
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Frequency of other examinations performed:
Mortality - recorded at 1, 3, 6, and 24 hours and daily thereafter until terminal sacrifice at day 14.
Clinical signs - the animals were observed for adverse effects and drug induced toxicity at 1, 3, 6, and 24 hours and daily thereafter following adminsitration.
The animals were sacrificed on the 14th day and observed for gross pathological organ changes.
Statistics:
The 24- hour and 14-day LD50 determination was calculated in accordance with the method of Miller and Tainter, Exp. Bio. and Medicine, 57, pp261-264, 1944.
Preliminary study:
No data.
Sex:
male
Dose descriptor:
LD50
Effect level:
2 440 mg/kg bw
Based on:
not specified
Mortality:
See Table 1.
Clinical signs:
See Table 2. Rats administered between 1.74 and 3.28 gm/kg bw of the test material exhibited respiratory depression, ataxia, loss of righting reflex, ptosis, piloerection, tremors, decreased locomotor activity and dose-related mortality.
Body weight:
See Table 3.
Gross pathology:
There were no outstanding gross pathological changes.
Other findings:
No data.

Table 1 - Survival for the acute oral toxicity study of MCTR-6 -75

Dose (gm/kg bw)

Hour

Day

Total

1

3

6

24

48

72

4

5

6

7

8

9

10

11

12

13

14

1.74

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

2.17

4

4

4

3

3

3

3

3

3

3

3

3

3

3

3

3

3

3

2.61

4

4

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3.04

4

4

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3.48

4

4

2

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Table 2 - Observations made following adminstration of the test substance.

Dose (gm/kg bw)

Animal # / sex

Hour

Day

1

3

6

24

48

72

4

5-14

1.74

1 /M

A,R,D,T,PT

Rr,R,PT,T

A,R,D,PT

D,P

D

 

 

 

2 /M

A,R,D,T,PT

Rr,R,PT,T

A,R,D,PT

D,P

 

 

 

 

3 /F

A,R,D,T,PT

Rr,R,PT,T

A,R,D,PT

D,P

D

D

D

 

4 /F

A,R,D,T,PT

Rr,R,PT,T

A,R,D,PT

D,P

D

 

 

 

Dose (gm/kg bw)

Animal # / sex

Hour

Day

1

3

6

24

48

72

4

5

6-14

2.17

1 /M

A,PT,R,D,T

Rr,T,R,PT

Rr,R,T,PT

PT,D,PT

D,P

D,P

D

 

 

2 /M

A,PT,R,D,T

Rr,T,R,PT

Death

-

-

-

-

-

-

3 /F

A,PT,R,D,T

Rr,R,PT

A,R,D,T,PT

D,A,R,P,PT

R,PT,D,P

PT,D,P

D,P

D

 

4 /F

A,PT,R,D,T

Rr,T,R,PT

A,R,D,T,PT

PT,D,P

D,P

D,

D

 

 

Dose (gm/kg bw)

Animal # / sex

Hour

Day

1

3

6

24

48

72

4

5

6-14

2.61

1 /M

Rr,R,T,PT

Death

-

-

-

-

-

-

-

2 /M

Rr,R,T,PT

Rr,R,T,PT

A,R,D,P,PT

A,R,P,D,PT

D,P,PT

D,P,PT

D,P

D

-

3 /F

Rr,R,T,PT

Rr,R,T,PT

Death

-

-

-

-

-

-

4 /F

Rr,R,T,PT

Death

-

-

-

-

-

-

-

Dose (gm/kg bw)

Animal # / sex

Hour

Day

1

3

6

24

48

72

4

5

6

7-14

3.04

1 /M

Rr,R,T,PT

Rr,R,T,PT

Rr,R,PT

A,R,D,P,PT

R,PT,D,P

PT,D,P

D,P

D

D

-

2 /M

Rr,R,PT,T

Rr,R,PT,T

Death

 

 

 

 

 

 

 

3 /F

Rr,R,PT,T

Death

 

 

 

 

 

 

 

 

4 /F

Rr,R,PT,T

Death

 

 

 

 

 

 

 

 

Dose (gm/kg bw)

Animal # / sex

Hour

Day

1

3

6

24

48

72

4

5

6

7-14

3.48

1 /M

Rr,R,T,PT

Death

 

 

 

 

 

 

 

 

2 /M

Rr,R,PT,T

Rr,R,PT

Death

 

 

 

 

 

 

 

3 /F

Rr,R,PT,T

Death

 

 

 

 

 

 

 

 

4 /F

Rr,R,PT,T

Rr,R,PT

Death

 

 

 

 

 

 

 

A=Ataxia

D=Decreased locomotion

PT=Ptosis

R=Respiratory Depression

T=Tremors

Rr=Loss of righting reflex

Table 3 - Individual bodyweights

Dose (gm/kg bw)

Animal # / sex

Test day

0

7

14

1.74

1 /M

210

225

275

2 /M

210

235

280

3 /F

200

210

215

4 /F

205

210

225

2.17

1 /M

230

250

295

2 /M

225

-

-

3 /F

215

220

230

4 /F

220

215

225

2.61

1 /M

200

-

-

2 /M

210

220

245

3 /F

200

-

-

4 /F

200

-

-

 

3.04

 

1 /M

210

225

235

2 /M

215

-

-

3 /F

200

-

-

4 /F

210

-

-

3.48

1 /M

220

-

-

2 /M

215

-

-

3 /F

210

-

-

4 /F

210

-

-

Interpretation of results:
GHS criteria not met
Conclusions:
Male and female rats, when administered oral dose levels between 1.74 and 3.48 gm/kg bw of the test material exhibited respiratory depression, ataxia, loss of righting reflex, ptosis, piloerection, tremors, ecreased locomotor activity and dose-related mortality. Autopsies revealed no outstanding gross patholigical organ changes. The acute oral LD50 at 24 hours was 2.44 ± 0.19 gram/kg bw.
In accordance with Regulation (EC) No. 1272/2008 tetrapotassium triphosphate is not considered to be classified via the oral route.
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
no data
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
The diluted compound was fed by stomach tube to Sprague-Dawley albino rats. After the approximate Minimal Lethal Dose was determined, groups of rats were fed at levels designed to blanket the toxicity range thereby supplying data for calculation of the LD50 according to the method of Bliss. Observations were made for toxic symptoms and the viscera of animals that succumbed were examined macroscopically.
GLP compliance:
no
Remarks:
study predates GLP
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
no data
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: the test material was diluted to produce a 50.0% aqueous solution.
- Amount of vehicle (if gavage): no data
- Justification for choice of vehicle: no data
- Lot/batch no. (if required): not applicable
- Purity: not applicable

MAXIMUM DOSE VOLUME APPLIED: 5010 mg/kg bw

- Rationale for the selection of the starting dose: The minimum lethal dose was predetermined and used to provide a range of doses from which the LD50 could be calculated
Doses:
2510 mg/kg bw
3160 mg/kg bw
3980 mg/kg bw
5010 mg/kg bw
No. of animals per sex per dose:
21 animals grouped as follows:
2510 mg/kg bw - 3 female, 2 male
3160 mg/kg bw - 3 female, 3 male
3980 mg/kg bw - 2 female, 2 male
5010 mg/kg bw - 3 female, 2 male
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: no data
- Frequency of observations and weighing: no data
- Necropsy of survivors performed: yes
- Other examinations performed: observationf for toxic symptoms and the viscera of the animals that succumbed were examined macroscopically.
Statistics:
Calculation of the LD50 was performed from the data derived from the four dose groups accroding to a modified version of the method of Bliss.
Preliminary study:
no data
Sex:
male/female
Dose descriptor:
approximate LD50
Effect level:
3 160 mg/kg bw
Remarks on result:
other: The upper and lower limits of the LD50 were placed at 2685 to 3665 mg/kg bw
Mortality:
The deaths of the animals are recorded in the results table.
Survival time was several hours to four days with most deaths occuring overnight.
Clinical signs:
Toxic symptoms included weakness in a few hours followed by collapse and coma.
Body weight:
no data
Gross pathology:
At autopsy there was renal and pulmonary congestion by macroscopic examination.
Other findings:
no data

The Oral LD50 of Tetrapotassium Pyrophosphate for Rats

sample fed as a 50.0% aqueous solution

Animal No. – Sex

Weight Gm.

Dose mg/kg

Fate

1 – female

175

2510

survived

2 – male

200

2510

survived

3 – female

185

2510

died

4 – female

190

2510

survived

5 – male

210

2510

survived

 

 

 

 

6 – male

200

3160

died

7 – male

195

3160

survived

8 – female

175

3160

died

9 – female

185

3160

died

10 – female

200

3160

survived

11 – male

180

3160

survived

 

 

 

 

12 – male

190

3980

died

13 – female

165

3980

died

14- female

180

3980

survived

15 – male

175

3980

died

16 – male

200

3980

died

 

 

 

 

17 – male

195

5010

died

18 – female

170

5010

died

19 – female

185

5010

died

20 – female

175

5010

died

21 – male

190

5010

died

Under the author's system of classification the compound was classed as slightly toxic by oral ingestions in rats.

Interpretation of results:
GHS criteria not met
Conclusions:
The LD50 was placed at 3160 mg/kg bw and the test material was classified, by the author, as slightly toxic by oral ingestions in rats.

In accordance with Regulation (EC) No. 1272/2008 (EU CLP) tetrapotassium pyrophosphate is not considered to be classified for acute oral toxicity.
Endpoint conclusion
Quality of whole database:
LD50 >2,000 mg/kg bw
No key study exists for tetrapotassium pyrophosphate. A weight of evidence (reliability 1, 2 and 4 studies) on tetrapotassium pyrophosphate and the analogous substance tetrasodium pyrophosphate is used to meet the endpoint requirements.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
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.
Qualifier:
according to guideline
Guideline:
EPA OPP 81-3 (Acute inhalation toxicity)
Deviations:
yes
Remarks:
- minor deviations: the chamber and room humidity was slightly higher than recommended in the guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Remarks:
- minor deviations: the chamber and room humidity was slightly higher than recommended in the guideline
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
yes
Remarks:
- minor deviations: the chamber and room humidity was slightly higher than recommended in the guideline
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Kingston, NY
- Age at study initiation: The actual age of the rats was not specified, only that they were young adults.
- Weight at study initiation (± SD): Males: 318 ± 11.4 g; Females: 249 ± 12.6 g
- Fasting period before study: No data
- Housing: Animals were housed individually in stainless steel suspended rat cages. Deosorb bedding was used in the litter pans.
- Diet: Purina Laboratory Rodent Chow 5001 available ad libitum
- Water: Tap water available ad libitum
- Acclimation period: Minimum of 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 23 °C (quoted in the study as 69 - 74 °F)
- Humidity (%): 42 - 77%
- Air changes (per hr): 14.2 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 h fluorescent light and 12 h dark cycle
Route of administration:
inhalation: dust
Type of inhalation exposure:
whole body
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The Rochester type exposure chamber was made of stainless steel and glass and was operated dynamically. The calculated 99% equilibrium time for the chamber at a flow rate of 35.6 L per minute was 19.4 minutes (equivalent to 14.2 "air changes per hour").
- Exposure chamber volume: 150 L
- Method of holding animals in test chamber: The test animals were assigned to and housed in individual compartments of a wire mesh cage bank (all on the same horizontal level) during the exposure.
- Source and rate of air: Breathing grade compressed air was used and the total chamber air flow rate was 35.6 L/minute.
- Method of conditioning air: No data
- System of generating particulates/aerosols: The test material was generated using a BGI Wright Dust Feeder II. The test material was desiccated and packed into large dust cups. Breathing Grade compressed air was metered to the Wright dust feeder through teflon tubing by a Matheson® 605 rotameter with a metal float. Rotameter back pressure was controlled using a Matheson® 3104C regulator. The dust feeder back pressure was monitored using a Marshalltown® back pressure gauge. The test material was made airborne by the compressed air dispersing the material into the exposure chamber. The concentration of the test atmosphere was controlled by the delivery rate setting of the Wright dust feeder.
- Method of particle size determination: The samples were drawn through a Sierra 218 cascade impactor at 2.78 liters per minute. The aerodynamic particle size distribution was determined by gravimetric analysis of the amount of test material collected on the impactor stages and subsequent determination of the mass median aerodynamic diameter (MMAD), geometric standard deviation and other particle size parameters by logarithmic-probability plotting.
- Treatment of exhaust air: The chamber air was exhausted from the bottom of the chamber and passed through an orifice tube system which continuously monitored airflow and then through a commercial filter box. The filter box was connected to a line leading to additional filters and an exhaust fan on the roof. The exhaust operated at a flow rate of 35.6 liters per minute, creating a slight negative pressure in the chamber, which was considered to be the total chamber air flow rate. The entire exposure system and primary exhaust filter were contained in a fume hood.
- Temperature, humidity, pressure in air chamber: The mean temperature and relative humidity in the chamber were 22 °C (71 °F) and 66%, respectively. The pressure in the air chamber was not measured.


TEST ATMOSPHERE
- Brief description of analytical method used: The airborne concentration of the test material was determined gravimetrically.
- Samples taken from breathing zone: Yes - Chamber air samples were taken on glass fiber filters held in cassettes at approximately one hour intervals during the exposure to determine the airborne concentration of test material. The airborne concentration of the test material was determined gravimetrically by drawing a known amount of chamber air through the filter. The samples were taken from the center of the chamber directly over the animal exposure caging.
The difference between gravimetric and nominal concentration was attributed to sedimentation of larger particles and/or adhesion of the test material to surfaces in the exposure chamber.


VEHICLE
- Not applicable: The test material was administered as received and a vehicle was not used.


TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: The fraction of particles less than or equal to 1 µm in mass aerodynamic diameter, based on the log-probability graphs, ranged from 7.6 to 9.4%. The fraction of particles less than or equal to 10 µm in mass aerodynamic diameter, based on the log probability graphs, ranged from 72.3 to 76.1%. These results indicated the test material was respirable in size to the rat.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The MMADs ranged from 4.61 to 4.87 micrometers (µm) with geometric standard deviations ranging from 2.98 to 3.39. The MMAD represents the smallest size that could be achieved in this study. The material is hygroscopic causing the particles to agglomerate and/or adhere to surfaces inside the chamber. Several trials were initially performed with various generation schemes and the system which was ultimately chosen provided the best performance.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
Nominal concentration: 35.14 mg/L (maximum attainable concentration)
Gravimetric concentration: 0.58 ± 0.103 mg/L
No. of animals per sex per dose:
5 animals/sex
Control animals:
no
Details on study design:
- Duration of observation period following administration: 28 days
- Frequency of observations and weighing: Animals were observed for signs of toxicity and mortality every 15 mins during the first hour of exposure, hourly for the remainder of the exposure, upon removal from the chamber, at 1 h post-exposure, twice daily thereafter for 27 days and once on day 28. Individual body weights were recorded on days 0, 1, 2, 4, 7, 14, 21 and 28.
- Necropsy of survivors performed: Yes
- Other examinations performed: No data
Statistics:
No data
Preliminary study:
Not applicable
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 0.58 mg/L air (analytical)
Exp. duration:
4 h
Mortality:
See Table 1.
One female died on day 1 and one male died on day 14 post-exposure.
Clinical signs:
other: See Table 1. Clinical signs noted during the exposure included lacrimation, material on fur, oral discharge and squinting eyes. Incidence of clinical signs was highest at the removal from chamber observation. Signs gradually resolved during the study, how
Body weight:
See Table 2.
Most animals lost weight through day 4 of the study, then began to gain weight in a normal pattern. At termination all surviving animals exhibited increases in body weight over their day 0 values.
Gross pathology:
See table 3.
There were no gross internal lesions observed in any animal which survived to study termination. One male which died on day 14 had discoloured lungs with many light red nodules. This animal was also observed to have a corneal opacity in one eye.
Other findings:
No data

See attached file for Tables 1, 2 and 3.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the test material caused mortality in one male and one female Sprague Dawley rat when administered for four hours at a mean, maximum attainable chamber concentration of 0.58 mg/L. Based on this, the LC50 for sodium acid pyrophosphate is considered to be greater than 0.58 mg/L.
This study is considered to be acceptable for classification and labelling in accordance with Regulation (EC) No 1272/2008 (EU CLP) and as such sodium acid pyrophosphate is not considered to be acutely toxic via the inhalation route (EU CLP).
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
LC50 >1.1 mg/L (highest dose tested)
One key study (reliability 2) is available.

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16/06/1988 to 30/06/1988
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: FMC Non-Definitive Dermal Toxicity Protocol (Number 7)
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
see any additional information on methods.
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Hazleton Research Animals, Inc., Denver, Pennsylvania on May 18, 1988
- Age at study initiation: Young
- Weight at study initiation: 2.21 - 2.74 kg
- Housing: individually housed in stainless steel cages. DAGB cageboard bedding was used in the litter pans.
- Diet (e.g. ad libitum): ad libitum; Purina High Fiber Rabbit Chow 5326
- Water (e.g. ad libitum): ad libitum; fresh tap water
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°F): 67 - 74°F
- Humidity (%): 52 - 69 %
- Photoperiod (hrs dark / hrs light): 12 hour fluorescent light: 12 hour dark cycle.


IN-LIFE DATES: May 18, 1988 - 13/06/1988
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
The test material was weighed out onto a 4" by 4" 8-ply gauze pad. The gauze was held in place with hypoallergenic tape. The test site was occluded with impervious plastic sheeting. Immediately after dosing each animal was fitted with an everted plastic Elizabethan collar. The collars remained in place until termination of the study.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 24 hour after exposure the wrapping was removed. The test site was wiped cleaned with a gauze moistened with methanol and then rinsed with tap water.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Please see additional information on materials and methods.
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- The test material was moistened with physiological saline.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6: 3 female, 3 males each at 2000 mg/kg bw.
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14
- Frequency of observations and weighing: The animals were observed for mortality and clinical signs ( local irritation excluded) at approximately 3 hours after dosing and daily thereafter for 14 days. Bodyweights were taken on the day of dosing and again on days 7 and 14. A description of the local irritation was recorded on days 1, 3, 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Statistics:
no data
Preliminary study:
no data
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
None of the animals died during the study
Clinical signs:
All animals remained healthy durng the observation period.
Body weight:
See Table 1. All but one rabbit gained weight during the study.
Gross pathology:
No gross lesions were found.
Other findings:
Results for local irritation are reported in Table 2.

Table 1 - Individual bodyweights

Animal # / sex

Day 0

(kg)

Dose

(g)

Amount applied

(mg/cm2)

Day 7

(kg)

Day 14

(kg)

1 / male

2.21

4.4

42.7

2.27

2.42

2 / male

2.66

5.3

51.5

2.65

2.78

3 / male

2.25

4.5

43.7

2.27

2.36

Mean ± SD

2.37 ± 0.249

 

46.0 ± 4.82

2.40 ± 0.219

2.52 ± 0.227

 

 

 

 

 

 

4 / female

2.55

5.1

49.5

2.69

2.85

5 / female

2.74

5.5

53.4

2.40

2.64

6 / female

2.54

5.1

49.5

2.56

2.60

Mean ± SD

2.61 ± 0.113

 

50.8 ± 2.25

2.55 ± 0.145

2.70 ± 0.134

Table 2 - Local irritation

Animal # / sex

Day 1

Day 3

Day 7

Day 14

(kg)

1 / male

N

N

De

De

2 / male

Er

Er

Es

Es, De

3 / male

Ed, Ne, Cb

Ed, Ne, Cb

Ts, Ne,Cb

Ne, Es, Ex, Tb, Ts

 

 

 

 

 

4 / female

Er

Er

Es, Tb

Es

5 / female

Er

N

N

N

6 / female

N

N

Es, Ts, Tb

Es, Ex

N - normal

De - desquamation

Er - Erythema

Ed - Edema

Ne - Necrosis

Cb - Chemcially burned

Ts - Skin thickening

Ex - Exfoliation

Tb - Tissue bleeding

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the test material is approximated to be relatively non-toxic.

In accordance with Regulation (EC) No. 1272/2008 (EU CLP) tetrapotassium pyrophosphate is not considered to be acutely toxic via the dermal route.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
LD50 >2,000 mg/kg bw
One key study (reliability 1, OECD 402) is available for an analogous substance.

Additional information

READ ACROSS JUSTIFICATION:

Read across between the following sodium and potassium pyrophosphates;

 

-         disodium dihydrogenpyrophosphate

-         trisodium hydrogen pyrophosphate

-         tertrasodium pyrophosphate

-         tetrapotassium pyrophosphate

 

Can be justified on the following basis; All substance contain a pyrophosphate anion and either a sodium or a potassium cation.

 

The pyrophosphate ion is the simplest form of a condensed phosphate group. A condensed phosphate anion has one or several P-O-P bonds. As the group contains only two phosphate groups, both of the phosphorus ions are classified as “terminal phosphorus”. The pyrophosphate can undergo ionisation with loss of H+ from each of the two –OH groups on each P and therefore can occur in the -1, -2 -3 or -4 state. The degree of ionisation is dependent upon the associated cations and the ambient pH (if in solution). Therefore the above substances have a pyrophosphate anion that is likely to behave in a similar way.

 

 In addition, the sodium and potassium cations are key elements in various cellular processes their import and export over cell membranes is regulated via pore systems and usually tightly regulated. As such, the presence of such cations is not expected to have an impact on the toxicity of the substances detailed above therefore as the pyrophosphate anion is common the results of toxicity studies can be reliably read-across within the group.

Oral toxicity

Sufficient data exists to evaluate the acute oral toxicity of tetrapotassium pyrophosphate without the need for read across. However studies on analogous substances are included to support the conclusion that sodium and tetrapotassium pyrophosphates have a similar systemic toxicity profile. Tetrasodium pyrophosphate is considered to be classified as Category 4 acute oral toxicity however the LD50 has been found to lie near the limit of classification and as such this should not affect the read across proposed for other studies nor should it indicate that tetrapotassium pyrophosphate should be classified.

A total of fourteen oral toxicity studies are available on the inorganic phosphate group, five on disodium dihydrogenpyrophosphate (SAPP), five on tetrapotassium pyrophosphate (TKPP) and four on tetrasodium pyrophosphate (TSPP) with reported LD50 values ranging between >1000 mg/kg bw and 3770 mg/kg bw.

 

Reference

Test substance

LD50[mg/kg bw]

Reliability

Newell (1974)

SAPP/ TSPP

1800/2980

4

Newel (1974)

SAPP

2300

4

Birch (1973)

SAPP

3600

4

Bullock (1972)

SAPP

>1000

3

Zipf (1957)

SAPP

2650

4

Birch (1973)

TKPP

2980

4

Younger (1961)

TKPP

3160

4

Kamienski (1971)

TKPP

>1000

4

Parke (1975)

TKPP

2440

2

Parke (1975)

TKPP

<5000

2

Birch (1973)

TSPP

3770

4

Jones (1988)

TSPP

1825 (f)

2150 (m)

4

 Bradshaw (2010)  TSPP  >300 ≤ 2000 (f)  1

 

Out of the five studies conducted on tetrapotassium pyrophosphatethe following studies are not adequate for use as part of a weight of evidence approach:

- Kamienski (1971) only provides a LD50value of >1000 mg/kg bw. Based on the investigation of two dose levels, 1000 mg/kg bw (no mortality) and 4640 mg/kg bw (mortality of four out of four). Therefore, these results can not be used for classification.

- Parke (1975) only tested at 5000 mg/kg bw and observed complete mortality, this study, despite being assigned reliability two is also not appropriate for use in the classification and labelling as the limit test does not relate to the classification criteria as defined by Regulation (EC) No. 1272/2008 (EU CLP).

 

The weight of evidence for tetrapotassium pyrophosphate indicates that it should not be classified via the oral route and an LD50of >2000 mg/kg bw is applicable. No studies are available that suggest classification is necessary and therefore no further testing is required.

Dermal toxicity

Sufficient data exists to evaluate the acute dermal toxicity of tetrapotassium pyrophosphate without the need for read across. However studies on analogous substances are included to support the conclusion that sodium and tetrapotassium pyrophosphates have a similar systemic toxicity profile.

A total of ten dermal toxicity studies are available on this inorganic phosphate group, three on disodium dihydrogenpyrophosphate (SAPP), four on tetrapotassium pyrophosphate (TKPP) and three on tetrasodium pyrophosphate (TSPP) with reported LD50values ranging from >300 mg/kg bw to >7940 mg/kg bw.

 

Reference

Test substance

LD50[mg/kg bw]

Reliability

Bradshaw (2010)

SAPP

>2000

1

Birch (1973)

SAPP/TKPP/TSPP

>7940/>7940/>7940

4

Parent (1990)

SAPP/TSPP

>300/>300

4

Jones (1988)

TSPP

>2000

2

Freeman (1988)

TKPP

>2000

2

Kamienski (1971)

TKPP

>4640

4

Parke (1977)

TKPP

>5000

4

One study on tetrapotassium pyrophosphate (TKPP) is assigned a reliability 1. Two studies, one on tetrapotassium pyrophosphate (TKPP) and one on tetrasodium pyrophosphate (TSPP), are reliability 2. All other studies are assigned reliability 4. The reported LD50values are all greater than figures with no mortalities observed at the highest test dose (where reported). The studies performed by Parent (1990) on disodium dihydrogenpyrophosphate and tetrasodium pyrophosphate are not true dermal toxicity tests but irritation studies with LD50values >300 mg/kg bw, the highest dose tested and as such these are not suitable for use for classification and labelling. These studies (reliability 4) do not provide full reports and no details on mortality data or other observations are available.

 

As only one true dermal toxicity study is available for tetrapotassium pyrophosphate (Freeman C, 1988) , this is used as the key study with the additional data on all substances provided to support the conclusion of low systemic toxicity. As the key study reports that the dermal LD50is > 2000 mg/kg bw tetrapotassium pyrophosphate is not considered to be classified and no further testing is deemed necessary. This conclusion is supported by the additional data.

Justification for classification or non-classification

Acute oral toxicity: The weight of evidence indicates that the oral LD50of tetrapotassium pyrophosphate is greater than the classification limit of 2000 mg/kg bw and therefore in accordance with Regulation (EC) No. 1272/2008 (EU CLP) no classification is proposed.

Acute inhalation toxicity: The LC50of tetrapotassium pyrophosphate for inhalation is >1.10 mg/L, the greatest attainable concentration and therefore in accordance with Regulation (EC) No. 1272/2008 (EU CLP) classification is not justified.

Acute dermal toxicity: The dermal LD50 of tetrapotassium pyrophosphate in the Wistar rat was estimated to be > 2000 mg/kg bw and is therefore not classified according to Regulation (EC) No. 1272/2008 (EU CLP).