Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental start date: 27 September 2018 and Experimental completion date: 16 October 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
2001
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: crystal powder
Details on test material:
- Synonym (Trade Name): ADDITIVE® 9735
- CAS Number: 102-40-9
- Molecular Formula: C10H14O4
- Molecular Weight: 198 g/mol
- Appearance: White crystal powder
- Expiration date: 30 August 2019
- Purity: 100%

Test animals

Species:
rat
Strain:
Wistar
Remarks:
(RccHan™:WIST) strain
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: ENVIGO RMS (UK) Limited, Oxon, UK.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 12 weeks of age
- Weight at study initiation: the body weight variation did not exceed +/- 20% of the mean body weight of any previously treated animals.
- Fasting period before study and afetr dosing: One overnight fast immediately before dosing and for approximately 3 to 4 hours afetr dosing.
- Housing: Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with softwood flake bedding.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs 15 changes per hour light): 12 hours continuous light and 12 hours darkness.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
DMSO
Details on oral exposure:
For the purpose of the study the test item was freshly prepared, as required, as a solution in dimethyl sulfoxide. Dimethyl sulfoxide was used because the test item did not dissolve/suspend in distilled water or arachis oil BP.
The test item was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was stable for this duration.
No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg
Doses:
2000 mg/kg at a concentration of 200 mg/mL (dose volume: 10 mL)
No. of animals per sex per dose:
A single female rat was dosed at 2000 mg/kg and in the absence of mortality at a dose level of 2000 mg/kg, an additional group of animals was treated at 2000 mg/kg.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.
- Necropsy of survivors performed: yes
- Other examinations performed: external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained
- Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days.
- Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.


Statistics:
The following computerized system was used in the study:
Delta Controls – ORCAview (Version 3.4.0)

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal was killed for humane reasons, approximately 2¼ hours after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence.
Clinical signs:
Hunched posture was noted in four animals. Additional signs of systemic toxicity noted in two animals were decreased respiratory rate and or labored respiration and hypothermia. Incidents of ptosis, lethargy and red/brown staining around the snout were also noted in one animal. Incidents of systemic toxicity also noted in the animal that was humanely killed during the study were prostration, hypothermia, increased salivation, splayed gait, loss of righting reflex and diarrhea.
Surviving animals appeared normal 4 hours to 2 days after dosing.
Body weight:
Surviving animals showed expected gains in body weight over the observation period.
Gross pathology:
Abnormalities noted at necropsy of the animal that was humanely killed were patchy pallor of the liver, pale kidneys and clear liquid present in the stomach. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Any other information on results incl. tables

Individual Clinical Observations and Mortality Data

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

H

H

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0
Female

0

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-1
Female

HPt

HPt

HPtRd

HPtLRl
HoSs

H

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2
Female

0

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3
Female

WsSRlPr

WsSRlPr

PrHoRlSRr
(PrHoSRlRrD)X*

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


0=      No signs of systemic toxicity                      D =       Diarrhea                             H =  Hunched posture             Ho =     Hypothermia

L =     Lethargy                                                   Pr =       Prostration                         Pt =       Ptosis                        Rd =     Decreased respiratory rate

Rl =   Labored respiration                             Rr =      Loss of righting reflex      S =  Increased salivation          Ws =     Splayed gait

Ss =   Red/brown staining around the snout

X* =  One animal was killed for humane reasons, approximately 2¼ hours after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence

Individual Body Weights and Body Weight Changes

Dose Level

(mg/kg)

Animal Number and Sex

Body Weight (g) at Day

Body Weight (g)
at Death

Body Weight Gain (g) During Week

0

7

14

1

2

2000

1-0 Female

155

186

201

 

31

15

2-0 Female

156

170

190

 

14

20

2-1 Female

158

172

187

 

14

15

2-2 Female

151

170

183

 

19

13

2-3 Female

162

-

-

156

-

-

- =       Animal dead

Individual Necropsy Findings

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

Killed Day 14

No abnormalities detected

2-1 Female

Killed Day 14

No abnormalities detected

2-2 Female

Killed Day 14

No abnormalities detected

2-3 Female

Humanely killed Day 0

Liver: patchy pallor

Kidneys: pale

Stomach: clear liquid present

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System  Category 5).
The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures.
Executive summary:

Introduction

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

Methods

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 item, as asolutionindimethyl sulfoxide, at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

Results

Mortality. Oneanimal was killed for humane reasons, approximately2¼ hours after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence.  

Clinical Observations. Hunched posture was noted in all four animals. Additional signs of systemic toxicity noted in two animals were decreased respiratory rate and or labored respiration and hypothermia with incidents of ptosis, lethargy, prostration, hypothermia, increased salivation, splayed gait, loss of righting reflex, diarrhea and red/brown staining around the snout. Surviving animals appeared normal 4 hours to 2 days after dosing.

Body Weight. Surviving animals showed expected gains in body weight.

Necropsy. Abnormalities noted at necropsy of the animal that was humanely killed were patchy pallor of the liver, pale kidneys and clear liquid present in the stomach. 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 item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System-Category 5).

The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures.