Registration Dossier

Administrative data

Description of key information

 Single oral application of 2000 mg submission substance per kg body weight via gavage did not cause lethality in male and female Wistar-rats during the 14 day observation period, resulting in an oral LD50 > 2000 mg/kg bw. No data on acute toxicity after inhalation are available. The submission substance was evaluated for Acute Dermal Toxicity in Sprague Dawley Rats as per the OECD guideline for the testing of chemicals No. 402, “Acute Dermal Toxicity - Fixed Dose Procedure”. Under the experimental conditions employed and based on the above results, it is concluded that the acute dermal median lethal dose (LD50) of submission substance in Sprague Dawley rats is > 2000 mg/kg body weight. Reliable data from one guideline study on acute toxicity after dermal application is available for a surrogate of the submission substance. The LD50 value from this study is above 2000 mg/kg bw as well.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 17 APR 1996 to 07 MAY 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes
Remarks:
according to Principles of Good Laboratory Practice, annex of paragraph 19a, section 1 of the chemical law of July 25, 1994
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst Aktiengesellschaft, Kastengrund, SPF breeding colony, Germany
- Age at study initiation: male animals approximately 7 weeks; female animals approximately 8 weeks
- Weight at study initiation: males mean: 188 g; females mean: 162 g
- Fasting period before study: from about 16 hours before to 3 - 4 hours after treatment
- Housing: in fully air-conditioned rooms in macrolon cages (type 4) on soft wood granulate in groups of 5 animals
- Diet: ssniff R/M-H (V 1534), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least one day (breeding at identical conditions)

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22 +/- 3
- Humidity (%): 50 +/- 20
- Air changes (per hr): fully air conditioned rooms
- Photoperiod (hrs dark / hrs light):12/12
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
deionised
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20% suspension
- Amount of vehicle (if gavage): 10 mL/kg bw
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once.
- Frequency of and weighing: weekly
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no animal died during the 14-day observation period
Mortality:
No deaths occurred during the whole study.
Clinical signs:
The following clinical signs were observed after the application of the test material:
squatting posture, decreased spontaneous activity, stilted gait. The clinical symptoms had reversed 4 to 6 hours after application.
Body weight:
Development of body weight was not impaired.
Gross pathology:
The animals killed at the end of the observation period showed no macroscopically visible changes.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Single oral application of 2000 mg test substance per kg body weight via gavage did not cause lethality in male and female Wistar-rats during the 14 day observation period, resulting in a LD50 > 2000 mg/kg bw.
Executive summary:

10 Wistar-rats (5 males and 5 females) were subjected to test acute oral toxicity. The test substance was administered by gavage at a dose of 2000 mg/kg body weight (vehicle deionised water). Body weight development was not impaired, general clinical signs observed were reversible within 6 hours after substance application and there were no macroscopically visible changes found. No animal died during the 14 day observation period, resulting in a LD50 > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
The study is GLP compliant and has Klimisch score 1. Another reliable study (Klimisch 1 and GLP) performed with a surrogate of the submission substance supports the reported findings for the submission substance.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study 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:
02 May 2018 to 25 June 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD Guideline for Testing of Chemicals No. 402 (Section 4: Health Effects) “Acute Dermal Toxicity: Fixed Dose Procedure” adopted on 09 October 2017
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
OECD Guideline for Testing of Chemicals No. 402 (Section 4: Health Effects) “Acute Dermal Toxicity: Fixed Dose Procedure” adopted on 09 October 2017
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- batch No.of test material: 18-UH-0703
- Expiration date of the batch: March 2020


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29°C)

FORM AS APPLIED IN THE TEST (if different from that of starting material): As such based on individual animal body weight

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: In-house
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10 weeks
- Weight at study initiation: 201.16 g to 214.88 g
- Fasting period before study: no
- Housing: standard polypropylene cage (size: L 430 x B 285 x H 150 mm)
- Diet (e.g. ad libitum): Altromin maintenance diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG)
- Water (e.g. ad libitum): Deep bore-well water passed through Reverse osmosis unit
- Acclimation period: 04 May 2018 to 13 May 2018

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6°C to 22.9°C
- Humidity (%): 47% to 65%
- Air changes (per hr): 12 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours fluorescent light and 12 hours dark cycle

IN-LIFE DATES: From: 04 May 2018 To: 28 May 2018
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorso-lateral area of the trunk of the animals
- % coverage: approximately 10% of the total body surface
- Type of wrap if used: non-irritating adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed using distilled water and dried with absorbent cotton
- Time after start of exposure: 24 hours

Duration of exposure:
The contact period of test item was about 24 hours
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
Range finding study : 1 Female
Main study : 2 Females
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All the animals were observed for clinical signs of toxicity and mortality at 20 to 30 min, 1 hr (±10 mins), 2 hrs (±10 mins), 4 hrs (±10 mins) and 6 hrs (±10 mins) post dosing on Day 1 and thereafter once daily for clinical signs of toxicity and twice daily for mortality during the 14 days observation period.
Individual animal body weight was recorded at receipt, on day 1 before test item application and on day 8 and 15 during the experimental period.
- Necropsy of survivors performed: yes
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
No mortality
Clinical signs:
No clinical signs
Body weight:
No treatment related changes in body weight and percent change in body weight with respect to day 1 were noted. Normal increase in body weights were noted during the observation period
Gross pathology:
No treatment related gross pathological changes were noted at 2000 mg/kg body weight (range finding study and main study) during necropsy

TABLE 1.     CLINICAL SIGNS OF TOXICITY ANDMORTALITY RECORD

Phase of the Experiment

 

Dose (mg/kg body weight)

Animal No.

Sex

Time of Dosing

(AM)

Clinical Signs of Toxicity and Mortality on Day 1

Clinical Signs of Toxicity and Mortality on days

20-30

mins

1 hr

(±10 mins)

2 hrs

(±10 mins)

4 hrs

(±10 mins)

6 hrs

(±10 mins)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Range Finding Study

2000

Rc9161

F

10:50

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Main Study

2000

Rc9162

F

10:39

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rc9163

F

10:41

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

    N: Normal; F: Female; min: minutes; hr/hrs: hour/hours


TABLE 2.   BODY WEIGHT (g) AND PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1

Phase of the Experiment

Dose (mg/kg)

Animal No.

Sex

Body Weight (g) on Days

Percent Change in Body Weight with Respect to Day

1

8

15

1 to 8

1 to 15

Range Finding Study

2000

Rc9161

F

219.53

235.60

251.73

 

7.32

14.67

Main Study

2000

Rc9162

F

241.43

256.34

270.98

 

6.18

12.24

Rc9163

F

230.94

246.70

262.14

6.82

13.51

Mean

236.19

251.52

266.56

 

6.50

12.87

±SD

7.42

6.82

6.25

 

0.46

0.90

 

 

n

 

2

2

2

 

2

2

     F: Female; SD: Standard Deviation; n: Number of animals

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


TABLE 3.    GROSS PATHOLOGY FINDINGS

Phase of the Experiment

Dose

(mg/kg body weight)

Animal No.

Sex

Fate

Gross Pathology Findings

External

Internal

Range finding Study

2000

Rc9161

F

TS

NAD

NAD

Main Study

2000

Rc9162

F

TS

NAD

NAD

Rc9163

F

TS

NAD

NAD

 NAD: No Abnormality Detected; F: Female;TS: Terminal Sacrifice

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
Under the experimental conditions employed and based on the above results, it is concluded that the acute dermal median lethal dose (LD50) of test item 6-(Isononanoylamino) hexanoic acid, compound with 2,2′,2″-nitrilotriethanol (1:1) in Sprague Dawley rats is > 2000 mg/kg body weight and classified as “Category 5 / Unclassified” (2000 < ATE ≤ 5000 mg/kg body weight) according to the Globally Harmonized System (GHS) of Classification
Executive summary:

The test item 6-(Isononanoylamino) hexanoic acid, compound with 2,2′,2″-nitrilotriethanol (1:1) obtained from Clariant India Ltd.was evaluated for Acute Dermal Toxicity in Sprague Dawley Rats as per the OECD guideline for the testing of chemicals No. 402, “Acute Dermal Toxicity - Fixed Dose Procedure”.

The study was performed in two phases i.e. range finding study and main study. Range finding study was performed with one female rat and main study was performed with two female rats. On the day before the application of the test item, fur on the dorso-lateral area of the trunk of the animals was removed by clipping closely with an electric hair clipper and care was taken to avoid abrading the skin.

The required quantity of the test item was applied as uniform film over an area of approximately 10% of the total body surface. The test item was held on to the applied surface by covering with cotton gauze dressing and wrapped with non-irritating adhesive tape and finally the application site was wrapped using semi-occlusive crepe bandage. The contact period of test item was 24 hours. At the end of the contact period, the residual test item was washed using distilled water and dried with absorbent cotton.

Since the oral LD50of test item is >2000 mg/kg for rats as per material safety data sheet provided by the sponsor, a starting dose of 2000 mg/kg body weight was selected from the fixed dose levels of 50, 200, 1000 and 2000 mg/kg body weight. No clinical signs and mortalities were observed at the dose level of 2000 mg/kg body weight in range finding study. Hence, during main study, two animals were administered with the same dose level of 2000 mg/kg body weight. No clinical signs were observed at the dose level of 2000 mg/kg body weight. Hence, no further testing was carried out.

All the animals were observed for clinical signs of toxicity and mortality at 20 to 30 min, 1 hr (±10 mins), 2 hrs (±10 mins), 4 hrs (±10 mins) and 6 hrs (±10 mins) on treatment day 1 and thereafter once daily for clinical signs of toxicity and twice daily for mortality during the 14 days observation period. The body weight was recorded on day 1 before test item application and on day 8 and 15. At the end of observation period, all the animals were sacrificed under carbon dioxide anaesthesia and subjected to necropsy and detailed gross pathological examination.

No mortality, clinical signs and skin reactions were noted. No treatment related changes in body weight and percent change in body weight with respect to day 1 were noted. Normal increase in body weights were noted during the observation period. No treatment related gross pathological changes were noted at 2000 mg/kg body weight (range finding study and main study) during necropsy.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
The study is GLP compliant and has Klimisch score 1.

Additional information

10 Wistar-rats (5 males and 5 females) were subjected to test acute oral toxicity of the submission substance (OECD 401, GLP compliant). Within this limit test the test substance was administered by gavage at a dose of 2000 mg/kg body weight (vehicle starch mucilage). Body weight development was not impaired, general clinical signs observed were reversible within 8 hours after substance application and there were no macroscopically visible changes found. No animal died during the 14 day observation period, resulting in a LD50 > 2000 mg/kg bw.

The results for the submission substance were verfied by another study performed with a surrogate of the submission substance using the same study design (OECD 401, Limit test, GLP). As no irreversible effects were seen and no animal died during the 14 day observation period the resulting LD50 is > 2000 mg/kg bw for rats.

The submission substance was evaluated for Acute Dermal Toxicity in Sprague Dawley Rats as per the OECD guideline for the testing of chemicals No. 402, “Acute Dermal Toxicity - Fixed Dose Procedure”. A starting dose of 2000 mg/kg body weight was selected from the fixed dose levels of 50, 200, 1000 and 2000 mg/kg body weight. No clinical signs and mortalities were observed at the dose level of 2000 mg/kg body weight in range finding study. Hence, during main study, two animals were administered with the same dose level of 2000 mg/kg body weight. No mortality, clinical signs and skin reactions were noted. No treatment related changes in body weight and percent change in body weight with respect to day 1 were noted. Normal increase in body weights were noted during the observation period. No treatment related gross pathological changes were noted at 2000 mg/kg body weight (range finding study and main study) during necropsy. Under the experimental conditions employed and based on the above results, it is concluded that the acute dermal median lethal dose (LD50) of the submission substance in Sprague Dawley rats is > 2000 mg/kg body weight

An acute dermal toxicity study was performed with a surrogate of the submission substance in RjHan:(WI) Wistar rats, in compliance with OECD Guideline No. 402. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied as a single dermal 24-hour exposure followed by a 14-day observation period. Within this study the following results were obtained: No mortality occurred. No clinical signs were observed after the treatment with the test item or during the 14-day observation period and no local dermal signs were existing. The body weight and body weight gain of treated animals did not show any adverse effects. There was no evidence of the test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Taken from this observations a LD50 is > 2000 mg/kg body weight.


Justification for selection of acute toxicity – oral endpoint
Reliable study performed with the submission substance and a surrogate substance

Justification for selection of acute toxicity – dermal endpoint
Reliable study performed with a surrogate of the submission substance and a surrogate substance

Justification for classification or non-classification

The oral LD50 values for the submission substance as well as for a surrogate of the submission substance are > 2000 mg/kg bw, the limit for classification. No lethality was observed after single dermal application of ≥ 2000 mg submission substance and surrogate per kg bw. No data on acute toxicity after inhalation exposure are available.

The submission substance does not have to be classified for acute toxicity according to Regulation (EC) No 1272/2008 or to Council Directive 67/548/EEC.

Furthermore, the submission substance does not have to be classified for specific target organ toxicity – single exposure according to Regulation (EC) No 1272/2008, as no specific toxic effects were observed after acute exposure.