Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 September 2007 to 17 October 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

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 inspection: 30 August 2005 Date of Signature: 21 November 2005
Test type:
fixed dose procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 3-Octadecyloxypropyl-N,N,N-trimethylammonium chloride
- Molecular formula (if other than submission substance): Not applicable
- Molecular weight (if other than submission substance): Not applicable
- Smiles notation (if other than submission substance): Not applicable
- InChl (if other than submission substance): Not applicable
- Structural formula attached as image file (if other than submission substance): Not applicable
- Substance type: white solid
- Physical state: solid
- Analytical purity: Not reported
- Impurities (identity and concentrations): Not reported
- Composition of test material, percentage of components: Not reported
- Isomers composition: Not reported
- Purity test date: Not reported
- Lot/batch No.: Exp. I-070518
- Expiration date of the lot/batch: Not reported
- Radiochemical purity (if radiolabelling): Not applicable
- Specific activity (if radiolabelling): Not applicable
- Locations of the label (if radiolabelling): Not applicable
- Expiration date of radiochemical substance (if radiolabelling): Not applicable
- Stability under test conditions: Not reported.
- Storage condition of test material: room temperature in the dark
- Other: Not reported.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Age at study initiation: 8 to 12 weeks of age.
- Weight at study initiation: Weights not recorded, however, the bodyweights fell within an interval of +/- 20% of the mean initial bodyweight of any previously dosed animal(s).
- Fasting period before study: Overnight fasting immediately before dosing and for approximately three to four hours after 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): Certified Rat and Mouse Diet ad libitum. The diet was routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Water (e.g. ad libitum): Mains drinking water ad libitum. The water was routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Set to achieve limits of 19 to 25ºC.
- Humidity (%): Set to achieve limits of 30 to 70%.
- Air changes (per hr): At least 15 changes per hour.
- Photoperiod (hrs dark / hrs light): Twelve hours continuous light (06:00 to 18:00) and 12 hours darkness.

IN-LIFE DATES: From: Day 0 (the day of dosing) up to Day 14.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The substance was present at levels of 30 and 200 mg/ml in the vehicle for doses of 300 and 2000 mg/kg, respectively.

- Amount of vehicle (if gavage): Test material was made into a solution with the vehicle at a dose level of 10 ml/kg.

MAXIMUM DOSE VOLUME APPLIED:
10 ml/kg

CLASS METHOD (if applicable)
Not applicable.
Doses:
300 and 2000 mg/kg.
No. of animals per sex per dose:
Five animals at 300 mg/kg and 1 animal at 2000 mg/kg.
Control animals:
no
Details on study design:
- Duration of observation period following administration:
Day 0 (the day of dosing) up to Day 14.

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

- Necropsy of survivors performed: At the end of the observation period the surviving animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

- Other examinations performed: clinical signs, body weight, necropsy and mortality.
Statistics:
Not reported.

Results and discussion

Preliminary study:
In the absence of data regarding the toxicity of the test material, 300 mg/kg was chosen as the starting dose.
In the absence of toxicity at a dose level of 300 mg/kg, an additional animal was treated as follows,
One animal at 2000 mg/kg and 4 animals at 300 mg/kg.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Mortality:
The animal was killed in extremis 2 days after dosing. There were no deaths at 300 mg/kg.
Clinical signs:
Signs of systematic toxicity were noted 1 and 2 days after dosing were hunched posture, lethargy, pilo-erection, decreased respiratory rate, laboured respiration, diarrhoea, diuresis, hypothermia, ataxia, pallor of the extremities, ptosis and dehydration at 2000 mg/kg (Table 1).
No signs of systematic toxicity were noted during the observation period at 300 mg/kg.
Body weight:
Individual bodyweights and body weight changes for 2000 mg/kg are given in Table 2, and for 300 mg/kg in Table 3.
All animals at 300 mg/kg showed expected gains in bodyweight over the observation period.

The surviving animals showed expected gains in bodyweight over the study period.
Gross pathology:
No abnormalities were noted at 300 and 2000 mg/kg.
Other findings:
Not reported.

Any other information on results incl. tables

Table 1 Individual Clinical Observations and Mortality Data - 2000 mg/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

0

HLPRdRl
DDuHo

HLPPtE
DhRdRlADDuHoX*

 

 

 

 

 

 

 

 

 

 

 

 

0= No signs of systemic toxicity                     

A = Ataxia                                                            

D = Diarrhoea                                      

Dh = Dehydration

Du = Diuresis                                                       

E = Pallor of the extremities                               

H = Hunched posture                         

Ho = Hypothermia

L = Lethargy                                                        

P = Pilo-erection                                                  

Pt =  Ptosis

Rd= Decreased respiratory rate

Rl = Laboured respiration                                  

X* = Animal killed in extremis

Table 2 Individual Bodyweights and Bodyweight Changes - 2000 mg/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

218

-

-

189

-

-

  - = Animal dead

Table 3 Individual Bodyweights and Bodyweight Changes- 300 mg/kg

Dose Level

mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Gain (g) During Week

0

7

14

1

2

300

1-0 Female

249

268

273

19

5

3-0 Female

233

252

286

19

34

3-1 Female

215

247

280

32

33

3-2 Female

244

272

296

28

24

3-3 Female

218

223

252

5

29

Applicant's summary and conclusion

Interpretation of results:
harmful
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was estimated to be in the range of 300 ‑ 2000  mg/kg bodyweight. The test substance is classified as R22 Harmful if swallowed under Council Directive 67/548/EEC and Category 4 H302: Harmful if swallowed under Regulation (EC) No 1272/2008.
Executive summary:

Introduction. The study was performed to assess the acute oral toxicity of the test material in the Sprague‑Dawley CD 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 B1 Acute Toxicity (Oral) of Commission Directive 2004/73/EC

Method. Following a sighting test at dose levels of 300 mg/kg and 2000 mg/kg, a further group of four fasted females was given a single oral dose of test material, as a solution/emulsion 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.

Mortality. The animal treated at a dose level of 2000 mg/kg was killed in extremis two days after dosing.

Clinical Observations. There were no signs of systemic toxicity noted in animals treated at a dose level of 300 mg/kg. Hunched posture, lethargy, pilo-erection, decreased respiratory rate, laboured respiration, diarrhoea, diuresis, hypothermia, ataxia, pallor of the extremities, ptosis and dehydration were noted in the animal treated at a dose level of 2000 mg/kg.

Bodyweight. Surviving animals showed expected gains in bodyweight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion. The acute oral median lethal dose (LD50) of the test material in the female Sprague‑Dawley CD strain rat was estimated to be in the range of 300 ‑ 2000 mg/kg bodyweight. The test substance is classified as R22 Harmful if swallowed under Council Directive 67/548/EEC and Category 4 H302: Harmful if swallowed under Regulation (EC) No 1272/2008.