Polyamide wax

Endpoint summary

Description of key information

The acute oral, inhalation and dermal toxicity studies indicate that the substance is of low  toxicity if swallowed (rat LD0 > or = 2000 mg/kg bw), applied onto the skin (rat LD0 > or = 2000 mg/kg bw) or inhaled (rat LC0> or = 4060 mg/m3 ; maximum practicable concentration ).

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 2000-03-14 to 2000-06-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan U.K.
- Age at study initiation: 5 to 7 weeks
- Weight at study initiation: 97 to 107g
- Fasting period before study: overnight prior to and for approximately hours after dosing
- Housing: in metal cages polished stainless steel grid floors
- Diet (e.g. ad libitum): standard laboratory rodent diet ad libitum
- Water (e.g. ad libitum):water ad libitum
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 20.5°C
- Humidity (%): 33-50%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours of artificial light (0600-1800 hours) in each 24-hour period

Route of administration:

oral: gavage

Vehicle:

other: methylcellulose

Details on oral exposure:

The test substance was formulated at a concentration of 20% w/v in 1% w/v aqueous methylcellulose and administered by oral gavage using a plastic syringe and catheter at a dose volume of 10 ml/kg bw.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: soon after dosing, and at frequent intervals for the remainder of Day 1. On subsequent days :morning and afternoon
until day 15
- Frequency of weighing: day 1, 8 and 15
- Necropsy of survivors performed: yes

Statistics:

None

Sex:

male/female

Dose descriptor:

LD0

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured in a group of 6 rats at a dose level of 2000 mg/kg of bw

Clinical signs:

other: No clinical signs were observed in any animals throughout the study

Gross pathology:

No abnormalities were revealed at the macroscopic examination at study termination on Day 15

Other findings:

No other findings

Table 7.2.1/2: Individual and group mean bodyweights (g) in treated animals during the observation period

Dose mg/kg	Animal n° and sex	Bodyweight (g) at
		Day 1 *	Day 8	A	Day 15	B
2000	3F	98	137	(39)	158	(21)
	4F	98	135	(37)	153	(18)
	5F	97	138	(41)	163	(25)
	Mean	98	137	(39)	158	(21)
	6M	107	147	(40)	198	(51)
	7M	105	151	(46)	197	(46)
	8M	106	168	(62)	214	(46)
	Mean	106	155	(49)	203	(48)

*: Prior to dosis

Bodyweight gain shown in parenthesis

A: Weight gain on Day 8 B: Weight gain on Day 15

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 of the test substance was higher than 2000 mg/kg in rats.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute oral toxicity according to OECD 423 guideline and in compliance with Good Laboratory Practices.

Groups of 3 rats/sex were administered by gavage a single dose of the test substance at 2000 mg/kg bw. Examinations for mortality, clinical signs and body weight gain were performed during the 14-day observation period. All surviving animals were necropsied at the end of the observation period. No mortality and no clinical signs were observed throughout the study. body weight gain was not affected by treatment. At necropsy, macroscopic examination of main organs showed no abnormalities.

The acute oral combined LD50 was found greater than 2000 mg/kg b.w.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD0

Value:

2 000 mg/kg bw

Quality of whole database:

Study complete and sufficient to fulfill the endpoint requirements.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2000-03-15 to 2001-01-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River UK
- Age at study initiation: 7 and 8 weeks
- Weight at study initiation: 257 to 281g (Males) and 213 to 240g (females)
- Fasting period before study: no
- Housing: housed by sex, in groups of 5, in cages made of stainless steel sheet and wire mesh
- Diet (e.g. ad libitum): SDS rat and mouse diet ad libitum
- Water (e.g. ad libitum): water ad libitum
- Acclimation period: 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): max: 22.5°C, min: 21.5°C
- Humidity (%): max: 60%, min: 34%
- Air changes (per hr): at least 15 air changes per hour
- Photoperiod (hrs dark / hrs light): artificial light between 06:00 and 18:00 GMT daily

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: flow through nose only chamber. This system was an aluminium alloy cylinder with a volume of approximately 30 liters. The conditioned test atmosphere entered through a port at the top centre of the chamber and passed out through a port at the base section below the level of the rats.
- Method of holding animals in test chamber: Rats were held in polycarbonate tubes with their snouts protruding from the end of the tubes into the exposure chamber.
- Source and rate of air: A supply of clean, dry, neutralised air was connected to the generator and the supply pressure was adjusted to give a flow
rate of 20l/min.
- System of generating particulates/aerosols: Wright Dust Feed mechanism designed to produce and maintain atmospheres of dust by suspending material scraped from the surface of a compressed powder in a stream of dry air. The concentrations of dust were altered by changing the gear ratio (and therefore the speed of rotation of the compressed powder towards the scraper blade) of the mechanism.The output of the WDF was connected to the top inlet port of the chamber via the elutriation column which reduced by sedimentation the amount of non respirable particulate in the test atmosphere.
- Temperature, humidity, pressure in air chamber: Temperature was measured using an alcohol-in-glass thermometer and relative humidity was measured with an infrared water vapour analyser. The temperature and relative humidity were recorded at the start of exposure and then at 30-minute intervals during the 4-hour exposure. The mean chamber temperatures were within expected ranges (20.5 and 20.4 °C for controls and test group) as well as the relative humidity (32 and 34 °C for controls and test group). Pressure in air chamber was not reported.
- Method of particle size determination: Particle size analysis of generated atmospheres was performed using a 8-stage cascade impactor (Marple 298). Samples were collected twice during exposure (86 and 210 min). Samples were also collected from a vacant animal exposure port (animals breathing zone). The collection substrates and the backup filter were weighed before and after sampling and the weight of test item, collected at each stage, was calculated by this difference. The total amount collected for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than < 0.52, 0.93, 1.55, 3.50, 6.0, 9.80, 14.80 and 21.30 µm (aerodynamic diameter) was calculated. From this data, the Mass Median Aerodynamic Diameter (MMAD), and Geometric Standard Deviation (GSD) were calculated. The mean MMAD was 5.5 µm and the geometric standard deviation (GSD) was 1.99. The Mass Median Aerodynamic Diameter was slightly above the targeted range of 1 to 4 µm, it came from the characteristics of the test item and the possible formation of aggregates during the aerosol generation.
- Treatment of exhaust air: The exhaust airflow was calibrated and adjusted to produce a slightly negative pressure.

TEST ATMOSPHERE
- Brief description of analytical method used: A measured volume of air was drawn at a rate of 2 litres/minute from an unused exposure
port on the exposure chamber through a glass microfibre filter, mounted in an open face filter holder and measured using a wet-type gas meter. Five samples were collected at approximately hourly intervals after equilibration during the exposure. The filters were weighed before and after sampling.The difference in the pre- and post-sampling weights, divided by the volume of atmosphere sampled, was equal to the actual achieved test atmosphere concentration. The mean achieved test atmosphere concentration was 4.06 +/- 0.214 mg/L: it was the maximum practicable concentration.
- Samples taken from breathing zone: yes

VEHICLE
- none

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

<= 4 h

Concentrations:

Mean achieved atmosphere concentration: 4.06 mg/L ; Standard deviation: 0.214
This concentration was the maximum practicable.

No. of animals per sex per dose:

5/sex

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: intermittently f observed for signs of reaction to the test substance during exposure and at least twice daily throughout
the observation period. the clinical signs were recorded at the end of the chamber equilibration period, at 0.25, 0.5 and 1 hours then hourly intervals during the exposure, immediately after exposure and then at 1.0 and 2.0 hours after exposure.
- Frequency of weighing: at least twice during the week prior to exposure, immediately before exposure and weekly during the observation period.
- Necropsy of survivors performed: yes, at the end of the 14-day observation period, the rats were killed by intraperitoneal injection of pentobarbitone sodium and exsanguinated when clinically dead. All rats were subjected to a deatailed macroscopic examination.

Statistics:

None

Sex:

male/female

Dose descriptor:

LC0

Effect level:

> 4.06 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: maximum practicable concentration

Mortality:

There were no unscheduled deaths.

Clinical signs:

other: - During exposure: > Exaggerated breathing was evident in all treated rats from 30 minutes into exposure. Soiling of the fur with excreta was observed in all treated and control animals from 30 minutes and 1 hour during the exposure respectively and w

Body weight:

The mean bodyweight gains for treated rats were less than controls during the first week following exposure. Thereafter, the mean bodyweight gain
of female and male treated rats was similar to and less than the control values respectively.
There were no treatment-related effects on food consumption.

Gross pathology:

There were no treatment-related findings at the macroscopic examination .
There were no treatment-related effects on lungs weight.

Table 7.2.2/1: Body weights - individual and group mean values (g)

Group	Rat	Day of observation
		-6	-3	0	7	14
1M (control)	1	213	241	263	322	351
	2	213	240	257	304	330
	3	225	258	281	333	356
	4	224	254	277	325	367
	5	215	249	271	318	355
	Mean	218	248	270	320	352
2M (treated)	11	213	244	272	310	343
	12	221	248	264	298	328
	13	212	244	265	308	346
	14	216	240	266	294	323
	15	216	239	259	291	316
	Mean	216	243	265	300	331
1F (control)	6	200	217	227	325	247
	7	201	206	218	225	241
	8	199	210	212	225	239
	9	203	216	215	231	251
	10	209	222	231	240	262
	Mean	202	214	221	231	248
2F (treated)	16	205	209	213	219	224
	17	204	219	223	230	252
	18	202	206	217	229	242
	19	203	210	231	235	241
	20	204	214	217	218	231
	Mean	204	212	220	226	238

 

Interpretation of results:

GHS criteria not met

Conclusions:

No death occurred at the concentration of 4.06 mg/L which is the maximum practical atmosphere concentration.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute inhalation toxicity study according to OECD 403 guideline and in compliance with Good Laboratory Practice. One treated and one control group, each of five male and five female Sprague-Dawley rats were exposed by nose-only inhalation to the test substance for 4 hours at a concentration of 4.06 mg/l or air respectively. The concentration of 4.06 mg/l was the maximal practical atmosphere concentration.

Each animal was observed for mortality and clinical signs at hourly intervals during exposure, then one hour and two hours after exposure and at least twice a day during the 14-day observation period. Bodyweights were recorded before treatment then on day of exposure and weekly during the observation period. All surviving animals were necropsied at the end of the observation period.

No mortality occurred during the study. Clinical signs as exaggerated breathing was evident for all treated animals from 30 minutes during the exposure and immediatly post exposure, persisting for most animals to day 4. Gasping and noisy breathing were noted for some animals post-exposure, the latter sign persisting in 1 male to Day 4. The mean bodyweight gain for treated rats was less than controls during the first week following exposure. Thereafter, the mean bodyweight gain of female and male treated rats was similar to and less than control values respectively. There were no treatment-related findings at the macroscopic examination.

The 4 -hour inhalation LC50 was found to be greater than 4.06 mg/l (ie 4060 mg/m3).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC0

Value:

4 060 mg/m³ air

Physical form:

inhalation: dust

Quality of whole database:

Study complete and sufficient to fulfill the endpoint requirements.

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:

From 2000-03-23 to 2000-08-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan U.K.
- Age at study initiation: 8 to 11 weeks of age
- Weight at study initiation: 210 to 257 g
- Fasting period before study: No
- Housing: Individually in metal cages fitted with grid floors
- Diet (e.g. ad libitum): Standard laboratory rodent diet ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 21.5°C
- Humidity (%): 36 - 60% relative humidity
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hours of artificial light (0600 - 1800 hours GMT) in each 24-hour period

Type of coverage:

semiocclusive

Vehicle:

other: 1% w/v aqueous methylcellulose

Details on dermal exposure:

TEST SITE
- Area of exposure: Dorso-lumbar region
- % coverage: approximately 10% of the total body surface area
- Type of wrap if used: waterproof dressing


REMOVAL OF TEST SUBSTANCE
- Removal of dressing: 24h post-exposure
- Washing: at 24h post-exposure, with warm water


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4.0 ml/kg bw
- Concentration (if solution): 500 mg/ml
- Constant volume or concentration used: yes


VEHICLE
- Concentration (if solution): 1% w/v aqueous methylcellulose
- Lot/batch no. (if required): no data
- Purity: no data

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Ten Sprague-Dawley rats (five males and five females).

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: soon after dosing and on two occasions during the day (the day of study termination : only the morning)
- Frequency of weighing: on days 1, 8 and 15
- Necropsy of survivors performed: yes

Statistics:

None

Sex:

male/female

Dose descriptor:

LD0

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured in any animal throughout the study.

Clinical signs:

other: There were no evidence of a systemic response in any animal throughout the study.

Gross pathology:

No macroscopic abnormalities were observed for animals killed at study termination.

Other findings:

- Other observations:
Slight irritation (erythema with oedema up to grade 1) was notable in seven rats following removal of the dressings on Day 2. In the majority of instances, the skin irritation had resolved by Day 4, however, in two animals slight irritation persisted before finally resolving in both instances by either Day 6 or Day 11. In two animals, a localised response (spots/scabbing) was first notable from Day 5 and persisted in one animal through to study termination (Day 15).

Table 7.2.2/1: Number of animals dead and number with evident toxicity

Dose (mg/kg bw)	Conc. in vehicle	Mortality (# dead/total)			Time range of deaths (hours)	Number with evident toxicity(#/total)
		Male	Female	Combined		Male	Female	Combined
2000	500 mg/ml	0	0	0	0	0	0	0

Mean erythema score for all animals:

at 24 hours: 0.7

at 48 hours: 0.4

at 72 hours: 0.2

Mean oedema score for all animals:

at 24 hours: 0.7

at 48 hours: 0

at 72 hours: 0

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 of the test substance was higher than 2000 mg/kg bw in rats.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute dermal toxicity study according to OECD 402 guideline and in compliance with Good Laboratory Practices. A group of ten rats (five males and five females) received a single topical application of the test substance formulated in 1% w/v aqueous methylcellulose and administered at a dosage of 2000 mg/kg bw. The application site was covered by a semi-occlusive dressing for 24 hours. Skin was washed with water at the end of the 24 -hour exposure period.

Examinations for mortality, clinical signs and body weight gain were performed during the 14-day observation period.

On completion of the observation period, the animals were sacrificed and then submitted to a macroscopic post-mortem examination.

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals. Body weight was not affected by the treatment.

Slight irritation was notable in seven rats following removal of the dressings on Day 2. In the majority of instances, the skin irritation resolved by Day 4 excepted in two animals where slight irritation persisted up to Day 5 or Day 10. In two animals, localized response (spots/scabbing) was first notable from Day 5 and persisted up to the study termination on Day 15 for one animal.

At necropsy, there were no macroscopic findings related to the test item administration.

The acute dermal LD50 was found greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD0

Value:

2 000 mg/kg bw

Quality of whole database:

Study complete and sufficient to fulfill the endpoint requirements.

Additional information

Oral route:

In an acute oral toxicity study performed according to OECD 423 guideline and in compliance with Good Laboratory Practices, groups of 3 rats/sex were administered a single dose of 2000 mg/kg bw of the test substance by gavage.

Examinations for mortality, clinical signs and body weight gain were performed during the 14-day observation period. All surviving animals were necropsied at the end of the observation period. No mortality and no clinical signs were observed throughout the study. Body weight gain was not affected by treatment. At necropsy, macroscopic examination of main organs showed no abnormalities.

The acute oral combined LD50 was found greater than 2000 mg/kg b.w (McRae, 2000a).

 

Dermal route:

In an acute dermal toxicity performed according to OECD 402 guideline and in compliance with Good Laboratory Practices, the test substance formulated in 1%w/v aqueous methylcellulose was applied to the skin of five males and five females rats at a dosage of 2000 mg/kg bw.The application site was covered by a semi-occlusive dressing for 24 hours.

Examinations for mortality, clinical signs and body weight gain were performed during the 14-day observation period.

On completion of the observation period, the animals were sacrificed and then submitted to a macroscopic post-mortem examination.

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals. Body weight was not affected by the treatment.

Slight irritation was notable in seven rats following removal of the dressings on Day 2. In the majority of instances, the skin irritation resolved by Day 4 except in two animals where slight irritation persisted up to Day 5 or Day 10. In two animals,localized response (spots/scabbing) was first notable from Day 5 and persisted up to the study termination on Day 15 for one animal.

At necropsy, there were no macroscopic findings related to the test item administration.

The acute dermal LD50 was found greater than 2000 mg/kg bw (McRae, 2000b).

 

Inhalation:

In an acute inhalation toxicity performed according to OECD 403 guideline and in compliance with Good Laboratory Practice, one treated and one control group, each of five male and five female Sprague-Dawley rats were exposed by nose-only inhalation to the test substance for 4 hours at a concentration of 4.06 mg/l or air respectively. The concentration of 4.06 mg/l was the maximal practical atmosphere concentration.

Each animal was observed for mortality and clinical signs at hourly intervals during exposure, then one hour and two hours after exposure and at least twice a day during the 14-day observation period. Bodyweights were recorded before treatment then on day of exposure and weekly during the observation period. All surviving animals were necropsied at the end of the observation period.

No mortality occurred during the study. Clinical signs as exaggerated breathing was evident for all treated animals from 30 minutes during the exposure and immediatly post exposure, persisting for most animals to day 4. Gasping and noisy breathing were noted for some animals post-exposure, the latter sign persisting in 1 male to Day 4. The mean bodyweight gain for treated rats was less than controls during the first week following exposure. Thereafter, the mean bodyweight gain of female and male treated rats was similar to and less than control values respectively. There were no treatment-related findings at the macroscopic examination.

The 4 -hour inhalation LC50 was found to be greater than 4.06 mg/l (ie 4060 mg/m3) (Paul, 2000).

 

Justification for classification or non-classification

No mortalities were noted in rats either after treatment by oral or dermal routes with a single dose of 2000 mg/kg body weight or exposure by inhalation for 4 hours to 4.06 mg/L which was the maximum practicable concentration.

On the basis of these results and according to regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures, no classification is warranted with respect to acute oral, dermal and inhalation toxicity.