N-acetylhexanelactam

Endpoint summary

Description of key information

The acute oral toxicity capacity of N-Acetylhexanelactam was tested in an OECD 401 guideline study.

The acute dermal toxicity capacity of N-Acetylhexanelactam was tested in an OECD 405 guideline study.

The test substance is of low acute toxicity with an LD50 oral of 1836 mg/kg bw and an LD50 dermal of > 2000 mg/kg bw.

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:

1998-09-08 to 1998-09-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline conform study report, GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Remarks:

Statement

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Doses:

1600, 2000 and 2500 mg/kg bw

No. of animals per sex per dose:

at 1600 mg/kg bw: 5 males/ 5 females
at 2000 mg/kg bw: 5 females
at 2500 mg/kg bw: 5 females

Control animals:

no

Statistics:

The LD50 and the equation of the probitline were established in female animals on the basis of the letahity rates by probit analysis.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 836 mg/kg bw

Based on:

test mat.

Mortality:

Under the conditions described the following lethality rates were observed:
males:
1600 mg/kg bw: absolute 0/5 males; relative: 0%
females:
1600 mg/kg bw: absolute 2/5 females; relative: 40%
2000 mg/kg bw: absolute 2/5 females; relative: 40%
2500 mg/kg bw: absolute 5/5 females; relative: 100%

Clinical signs:

other: Clinical sighn observed were: hypoactivity, stupor, sunken flanks, bristled coat, stilted gait, uncoordinated gait, ataxia, irregular respiration, narrowed palpebral fissures, prone position, absent paw reflex to pinching, absent righting reflex and narco

Gross pathology:

Macroscopic findings of animals found dead
Liver: light discoloration;
Stomach: detachment of the mucous membrane
Small intestine: diffuse reddening
Lung: discoloured red.
Animals sacrified at the scheduled end of observation period showed no macroscopically visible changes.

Under the conditions described the following lethality rates were observed:

males:

1600 mg/kg bw: absolute  0/5 males; relative: 0%

females:

1600 mg/kg bw: absolute  2/5 females; relative: 40%

2000 mg/kg bw: absolute  2/5 females; relative: 40%

2500 mg/kg bw: absolute  5/5 females; relative: 100%

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral median lethal dose (LD50) in female Sprague Dawley rats was found to be 1836 mg/kg bw.

Executive summary:

In an OECD 401 study the acute oral median lethal dose (LD50) of the test substance to the Sprague Dawley strain rat was found to be 1836 mg/kg bw. Doses of 1600, 2000 or 2500 mg/kg bw in sesame oil were applied once to the rats by gavage. Whereas 1600 mg/kg bw was applied to 5 male and 5 female rats, the remaining two doses were applied to 5 female rats each. Mortality occured in 5/5 female rats at 2500 mg/kg bw, in 2/5 females at 2000 mg/kg bw and in 2/5 females at 1600 mg/kg bw. No death occurred in male rats at 1600 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 836 mg/kg bw

Quality of whole database:

The study is an GLP study rated Klimisch 1.

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:

2017-06-28 to 2017-08-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

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)

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

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

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:

other: There were no clinical signs observed during the study.

Gross pathology:

No abnormality was detected at necropsy.

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)	8^th   day	Weight change (day 8 – Initial)	15^th 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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study is an GLP study rated Klimisch 1.

Justification for classification or non-classification

For the oral toxicity, the test substance has to be classified Category 4, according to Regulation (EC) No 1272/2008 of the European Parliament and of the council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006.

For the dermal toxicity, the test substance has not to be classified, according to Regulation (EC) No 1272/2008 of the European Parliament and of the council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006.

In accordance with column 2 of REACH Regulation (EC) No 1907/2006 Annex VIII no further testing for inhalation toxicity is required taking the vapor pressure into account and when the exposure of humans via inhalation is not likely and not a relevant route of exposure. The vapour pressure of the test substance is very low and inhalation is not exposure relevant as the test substance is manufactured in a closed system. PPE is obligatory and mandatory. No study on inhalation was performed with N-Acetylhexanelactam.