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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-06-28 to 2017-08-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
Guideline for Testing of Chemicals, Section 4, No. 402, “Acute Dermal Toxicity”, adopted: 24th February 1987.
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
N-acetylhexanelactam
EC Number:
217-565-6
EC Name:
N-acetylhexanelactam
Cas Number:
1888-91-1
Molecular formula:
C8H13NO2
IUPAC Name:
N-acetylhexanelactam
Test material form:
liquid
Details on test material:
Batch: not yet known
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Clariant Iberica Production, S.A., Tarragona Site, 43110 La Canonja, Spain

- Batch No.of test material: ESD0024613
- Expiration date of the batch: 06 May 2016
- Purity as per Certificate of Analysis: After 06 May 2018

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

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Geniron Biolabs Pvt. Ltd., No.93, Solur, Anekal - Thally Road, Anekal, Bengaluru – 562106, India

- Females (if applicable) nulliparous and non-pregnant: [yes]
- Age at study initiation: 8 to 9 weeks
- Weight at study initiation: Males : 272.1 to 285.3 g; Females: 229.8 to 241.3 g

- Housing: Animals were housed individually in standard polysulfone cages
(Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottle. Additionally, polycarbonate rat huts were placed inside the cage as an enrichment object and were changed along with the cage once a week.

- Diet (e.g. ad libitum): Hypro rat & mice pellet feed, manufactured by Pranav Agro Industries Ltd., Pune 411 030, Maharashtra, India, was provided to animals.

- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd, Mumbai 400 001, India, was provided to animals in polycarbonate bottles with stainless steel sipper tubes.

- Acclimation period: Start : 30 June 2017 End : 05 July 2017 (6 days)


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 64 to 67 %
- Air changes (per hr): 13.7 to 13.9 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 30 June 2017 To: 20 July 2017

Administration / exposure

Type of coverage:
semiocclusive
Details on dermal exposure:
TEST SITE
- Area of exposure: approximately 10 x 8 cm
- % coverage: 10% of the body surface
- Type of wrap if used: The applied area was covered with cotton gauze (size: Males: 9 x 6 cm; Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): after 24 hours
- Time after start of exposure:10.46 AM to 10.56 AM

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Refer Section 8.2.2.4 in Main Report
- Concentration (if solution): Not applicable
- Constant volume or concentration used: no
- For solids, paste formed: no

Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males and 5 females
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: On day 1, 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no pre-terminal deaths (mortality) observed during the study.
Clinical signs:
There were no clinical signs observed during the study.
Body weight:
All rats gained body weight throughout the observation period.
Gross pathology:
No abnormality was detected at necropsy.

Any other information on results incl. tables

Table 1.         Individual body weight, body weight changes and pre-terminal deaths

Group and

Dose

(mg/kg

body weight)

Rat

No.

S

e

x

Body weight (g)

No.dead /

No.tested

Pre- terminal deaths (%)

Initial

(at treatment)

8th  

day

Weight change

(day 8 – Initial)

15th

day

Weight change

(day 15 – Initial)

 

G1 and

2000

(1.82 mL/kg*)

Limit test

Rm6381

M

274.8

287.0

12.2

296.3

21.5

 

0/10

 

0

Rm6382

M

278.1

295.1

17.0

307.9

29.8

Rm6383

M

285.3

303.5

18.2

312.4

27.1

Rm6384

M

272.1

288.3

16.2

296.8

24.7

Rm6385

M

281.4

299.7

18.3

306.5

25.1

Rm6386

F

241.3

250.8

9.5

259.4

18.1

Rm6387

F

235.2

245.2

10.0

252.6

17.4

Rm6388

F

236.1

243.9

7.8

249.9

13.8

Rm6389

F

229.8

240.4

10.6

248.5

18.7

Rm6390

F

239.3

248.8

9.5

257.1

17.8

F: Female                 M: Male   

*: Calculated based on the density of the test item: 1.0995 g/cm3at 20° C (as per details provided by the sponsor).

Table 2.              Individual test item application, clinical signs and necropsy findings

Group and Dose

(mg/kg body weight)

Date &

Time of

Application

Rat

No.

Sex

Body weight initial

(g)

Volume

 applied

(mL)

 

Day of Observations

Day 1

2

3

1

hour

2

hours

3

hours

4

hours

 

 

AM

PM

AM

PM

G1 and

2000

(1.82 mL/kg*)

Limit test

06 July 2017

&

10.46 AM

to

10.56 AM

Rm6381

M

274.8

0.50

N

N

N

N

N

N

N

N

Rm6382

M

278.1

0.51

N

N

N

N

N

N

N

N

Rm6383

M

285.3

0.52

N

N

N

N

N

N

N

N

Rm6384

M

272.1

0.50

N

N

N

N

N

N

N

N

Rm6385

M

281.4

0.51

N

N

N

N

N

N

N

N

Rm6386

F

241.3

0.44

N

N

N

N

N

N

N

N

Rm6387

F

235.2

0.43

N

N

N

N

N

N

N

N

Rm6388

F

236.1

0.43

N

N

N

N

N

N

N

N

Rm6389

F

229.8

0.42

N

N

N

N

N

N

N

N

Rm6390

F

239.3

0.44

N

N

N

N

N

N

N

N

M: Male; F: Female; N: Normal; AM: Ante Meridian; PM: Post Meridian    

Table 2 contd.         Individual test item application, clinical signs and necropsy findings

Group and Dose

(mg/kg body weight)

Rat

No.

Sex

Day of Observations

Necropsy

Findings

4

5

6

7

8

9

10

11

12

13

14

15

G1 and

2000

(1.82 mL/kg*)

Limit test

Rm6381

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6382

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6383

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6384

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6385

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6386

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6387

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6388

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6389

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm6390

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

M: Male; F: Female; NAD: No Abnormality Detected; N: Normal; AM: Ante Meridian;

PM: Post Meridian

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the present study results, single dermal application of the test item was associated with no mortality and neither signs of toxicity nor signs of irritation.

The acute dermal LD50 of N-acetylhexanelactam is more than 2000 mg/kg body weight in male and female Wistar rats.
Executive summary:

The acute dermal toxicity ofN-Acetylhexanelactamwas tested in 5 male and 5 female Wistar rats at the limit dose of 2000 mg/kg body weight.As per the Sponsor’s request initial limit dose of 2000 mg/kg body weight was applied to the 5 male and 5 female rats simultaneously. Based on the individual body weight, the undiluted test item at dose of 2000 mg/kg body weight and dose volume was 1.82 mL/kg body weight was calculated based on the density of the test item i.e., 1.0995g/cm3at 20°C(as per details provided by the sponsor). The undiluted test item was applied (semi-occlusive) directly to the clipped skin of the rat to cover about 10% of body surface of the rat. Then the cotton gauze was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was 24 hours.After the 24 hour contact period, the adhesive tape and cotton gauze were removed and the application area was washed with water and wiped dry using a clean towel. 

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. There were no clinical signs of toxicity and mortality. All rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy. There were no abnormalities detected at necropsy.

Based on the present study results, single dermal application of the test item was associated with no mortality and neither signs of toxicity nor signs of irritation.

The acute dermal LD50of N-Acetylhexanelactam is more than 2000 mg/kg body weight in male and female Wistar rats.