Calcium bis[4-[[3-[[2-hydroxy-3-[[(4-methoxyphenyl)amino]carbonyl]-1-naphthyl]azo]-4-methylbenzoyl]amino]benzenesulphonate]

Endpoint summary

Description of key information

Acute oral toxicity: 

Female Wistar-rats were subjected to test acute oral toxicity. The test
substance was administered by gavage at a dose of 10000 mg/kg bw (25% in demineralized water). No animal died during the 14 day observation period, resulting in a LD50 >10000 mg/kg bw.

LD50, oral: >10000 mg/kg bw

Acute dermal toxicity: 

Single dermal application of the test item to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity. The dermal LD50 was determined to be > 2000 mg / kg body weight.

LD50, dermal: >2000 mg/kg bw, RA from source substance PR 170

 

Acute inhalation toxicity:

Male and female Sprague Dawley rats were subjected to test acute inhalation toxicity. The test
substance was administered for 4 h at a dose of 5.04 mg/L air. No animal died during the 14 day observation period, resulting in a LC50 > 5.04 mg/L air.

LC50, inhalation > 5.04 mg/L air

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: WISKf(SPF71), Hoechst AG, breeding colony
- Weight at study initiation: 172 g +/- 3.39 g
- Fasting period before study: 16 h
- Housing: grouped, plastic cages
- Diet: Altromin 1324 (Altromin GmbH, Lage/Lippe, Germany), ad libitum
- Water: tap water, ad libitum

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

demineralised

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25 %

Doses:

10000 mg/kg bw

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of weighing: weekly
- Necropsy of survivors performed: yes

Key result

Sex:

female

Dose descriptor:

LD0

Effect level:

> 10 000 mg/kg bw

Based on:

test mat.

Mortality:

- no deaths occurred

Clinical signs:

other: - no animal showed any clinical symptoms - faeces was red-coloured

Gross pathology:

- animals killed at the end of the observation period showed no macroscopically visible changes

Interpretation of results:

not classified

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Single application of 10000 mg test substance per kg bw did not cause lethality in female Wistar-rats during the 14 day observation period, resulting in a LD50 > 10000 mg/kg bw.

Executive summary:

Female Wistar-rats were subjected to test acute oral toxicity. The test substance was administered by gavage at a dose of 10000 mg/kg bw (25% in demineralized water). No animal died during the 14 day observation period, resulting in a LD50 >10000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

10 000 mg/kg bw

Quality of whole database:

adequate reliability
LD50: >10000 mg/kg bw

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:

10 November 2022 to 01 December 2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

OECD Guidelines for Testing of Chemicals No. 403 – Traditional Protocol “Acute Inhalation Toxicity” adopted on 07 September 2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

traditional 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: In-house bred
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 weeks
- Weight at study initiation: Males: 182.12 g to 191.26 g; Females: 166.18 g to 179.49 g

- Housing: Three animals per sex were housed in a standard polycarbonate cage (size: L 430 × B 280 × H 210 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube
- Diet (e.g. ad libitum): yes and Altromin maintenance diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG)
- Water (e.g. ad libitum): yes and Deep bore-well water passed through reverse osmosis unit
- Acclimation period: 11 November 2022 to 16 November 2022

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9°C to 22.6°C
- Humidity (%): 43% to 66%
- 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: 11 November 2022 To: 01 December 2022

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

> 2.98 - <= 3.23 µm

Geometric standard deviation (GSD):

> 2.71 - <= 2.77

Remark on MMAD/GSD:

within the specified range as per guideline

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: nose-only dynamic inhalation exposure system
- Exposure chamber volume: 0.76
- Method of holding animals in test chamber: Restrainers
- Source and rate of air (airflow): 20 lt per minute
- Method of conditioning air: filtered air
- System of generating particulates/aerosols: Rotating Brush Generator (Palas RBG 1000 - supplied by Palas GmbH)
- Method of particle size determination: Gravimetric method
- Treatment of exhaust air: NaOH
- Temperature, humidity, pressure in air chamber: 22.3°C to 22.8°C, 52.9% to 56.5%, 60 psi

TEST ATMOSPHERE
- Samples taken from breathing zone: yes
- Time needed for equilibrium of exposure concentration before animal exposure:
t95 (min) = 3× Chamber Volume / Chamber airflow
= 3× 0.76 L / 20 L/min
= 0.11 minutes


CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: As per guidelines the limit concentration was 5 mg/L of air.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

> 0 - <= 240 min

No. of animals per sex per dose:

Limit Test - 3 Males and 3 Females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: once daily for clinical signs and twice daily for mortality and weighing of animals on day 1, 3, 7 and 14
- Necropsy of survivors performed: yes

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.04 mg/L air

Based on:

test mat.

Mortality:

No mortality

Clinical signs:

other: No treatment related clinical signs and mortality were observed at the mean limit concentration of 5.04 mg/L of air. Post exposure observation, i.e., on day 2 and day 3 test item color feces was noted

Body weight:

No treatment related changes were observed in body weight and percent change in body weight with respect to day 1 at the mean limit concentration of 5.04 mg/L of air. However, all animals showed slight decrease in body weight on day 2 due to exposure and increased in body weight from day 4 onwards

Gross pathology:

No treatment related gross pathological findings were observed at the mean limit concentration of 5.04 mg/L of air

• CLINICAL SIGNS AND MORTALITY RECORD

Group  & Concentration (mg/L of air)

Animal No.

Sex

Day 1

Days

During Exposure

Post exposure

1 hr*

2 hrs*

3 hrs*

4 hrs*

30-40 min

1 hr*

2#

3#

4

5

6

7

8

9

10

11

12

13

14

15

Limit Test & 5.04

Rh7081

M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rh7082

M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rh7083

M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rh7084

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rh7085

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rh7086

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

              *: ± 10 minutes; N: Normal; M: Male; F: Female; min: minute; hr(s): hour(s); #Test item color feces noted

 

                       

 

• BODY WEIGHT (g) AND PERCENT CHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1

Group &    Concentration (mg/L of air)	Animal No.	Sex	Body Weight (g) on Days						Percent Change in Body Weight with Respect to Day 1
			1^#	2	4	8	15		1-2	1-4	1-8	1-15
Limit Test & 5.04	Rh7081	M	203.36	200.65	204.77	217.17	239.19		-1.33	0.69	6.79	17.62
	Rh7082	M	211.19	209.39	213.39	228.24	252.32		-0.85	1.04	8.07	19.48
	Rh7083	M	208.93	205.49	209.42	223.46	246.25		-1.65	0.23	6.95	17.86
	Mean		207.83	205.18	209.19	222.96	245.92		-1.28	0.66	7.27	18.32
	(±) SD		4.03	4.38	4.31	5.55	6.57		0.40	0.40	0.70	1.01
	n		3	3	3	3	3		3	3	3	3
	Rh7084	F	182.41	180.82	183.58	192.61	211.67		-0.87	0.64	5.59	16.04
	Rh7085	F	187.05	186.66	188.15	199.82	214.70		-0.21	0.59	6.83	14.78
	Rh7086	F	195.88	193.71	196.21	206.09	223.52		-1.11	0.17	5.21	14.11
	Mean		188.45	187.06	189.31	199.51	216.63		-0.73	0.47	5.88	14.98
	(±) SD		6.84	6.45	6.39	6.75	6.16		0.47	0.26	0.84	0.98
	n		3	3	3	3	3		3	3	3	3

#: Prior to exposure; M: Male; F: Female; SD: Standard Deviation; n; Number of animals

• BREATHING ZONE CONCENTRATION (ACTUAL TEST ITEM CONCENTRATION)

 Technical Pre-test:

Sl. No.	Feed rate (mm/hour)	Rotation (rpm)	Initial weight (mg) (a)	Final weight (mg) (b)	Difference (mg) (c) = (b) – (a)	Air Flow Rate (L/min) (d)	Time (min) (e)	BZC (mg/L of air)	Mean BZC (mg/L of Air)
1	30	600	345.71	353.34	7.63	1.52	1	5.02	5.03
2	30	600	350.65	358.31	7.66	1.52	1	5.04

 Limit Test:

Sl. No.	Feed rate (mm/hour)	Rotation (rpm)	Initial weight (mg) (a)	Final weight (mg) (b)	Difference (mg) (c) = (b) – (a)	Air Flow Rate (L/min) (d)	Time (min) (e)	BZC (mg/L of air)	Mean BZC (mg/L of Air)
1	30	600	350.05	357.69	7.64	1.52	1	5.03	5.04
2	30	600	356.09	363.78	7.69	1.52	1	5.06
3	30	600	349.86	357.53	7.67	1.52	1	5.05

   BZC: Breathing Zone Concentration; Sampled volume: 1.52 L/min; Sampling time: 1 minute

 

BZC (f) =	Mass of test item collected on the filter paper (c)
	Volume of air passed through the filter paper (d) X time (e)

 

• CHAMBER (EXPOSURE) CONDITIONS

Technical Pre-test:

Concentration (mg/L of air)	SL. No.	Feed rate (mm/hour)	Rotation (rpm)	Temperature (°C)	Relative Humidity (%)	Oxygen Concentration (%)	Carbon dioxide Concentration (ppm)	Air Flow (L/min)
5.03	1	30	600	22.4	54.6	20.5	616	20
	2	30	600	22.3	53.5	20.6	625	20

 

Limit Test:

Concentration (mg/L of air)	SL. No.	Feed rate (mm/hour)	Rotation (rpm)	Temperature (°C)	Relative Humidity (%)	Oxygen Concentration (%)	Carbon dioxide Concentration (ppm)	Air Flow (L/min)
5.04	1	30	600	22.8	52.9	20.3	613	20
	2	30	600	22.4	54.5	20.6	621	20
	3	30	600	22.5	56.5	20.3	619	20
	4	30	600	22.3	55.8	20.4	615	20

Note: 1% Carbon dioxide = 10000 ppm

• SUMMARY OF CHAMBER (EXPOSURE) CONDITIONS

Technical Pre-Test
Chamber Conditions	Range
Feed Rate (mm/hour) and Speed (RPM)	30 and 600
Temperature (°C)	22.3 to 22.4
Relative humidity (%)	53.5 to 54.6
Oxygen concentration (%)	20.5 to 20.6
Carbon dioxide concentration (ppm)	616 to 625
Air Flow (L/min)*	20
BZC (mg/L)	5.02 to 5.04
MMAD (µm)	3.06 ^#
GSD	2.71 ^#
Limit Test
Feed Rate (mm/hour) and Speed (RPM)	30 and 600
Temperature (°C)	22.3 to 22.8
Relative humidity (%)	52.9 to 56.5
Oxygen concentration (%)	20.3 to 20.6
Carbon dioxide concentration (ppm)	613 to 621
Air Flow (L/min)*	20
BZC (mg/L)	5.03 to 5.06
MMAD (µm)	2.98 to 3.23
GSD	2.71 to 2.77

*: Values were constant throughout the exposure; ^#: Individual value; BZC: Breathing Zone Concentration (Actual Concentration), MMAD: Mass Median Aerodynamic Diameter, GSD: Geometric Standard Deviations.

 

 

• GROSS PATHOLOGICAL FINDINGS

Group & Concentration (mg/L of air)	Animal No.	Sex	Fate	Gross Pathology Findings
				External	Internal
Limit Test & 5.04	Rh7081	M	TS	NAD	NAD
	Rh7082	M	TS	NAD	NAD
	Rh7083	M	TS	NAD	NAD
	Rh7084	F	TS	NAD	NAD
	Rh7085	F	TS	NAD	NAD
	Rh7086	F	TS	NAD	NAD

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

Interpretation of results:

GHS criteria not met

Remarks:

Based on the results of this study the test item has not to be classified for acute inhalation toxicity according to Regulation (EC) No 1272/2008.

Conclusions:

Under the experimental conditions employed and based on the above results of experiment, there were no clinical signs and mortality observed at the mean limit concentration of 5.04 mg/L of air. Hence, the LC50 of the test item is > 5.04 mg/L of air.

Executive summary:

The test item was evaluated for acute inhalation toxicity in Sprague Dawley rats.

The objective of the study was to assess the toxic potential and to determine the LC₅₀ of test item when administered by inhalation route through flow-past nose-only dynamic inhalation equipment for a single 4 hours exposure to rats. Three male and three female rats were used for conducting the acute inhalation toxicity study.

As such test item was used during technical pre-test and limit test to generate the dust aerosols through rotating brush generator. The technical pre-test was carried out without animals. During the technical pre-test, the target concentration i.e.  5.04 mg/L of air was achieved at the feed rate of 30 mm/hour with a rotation of 600 RPM.

During the exposure period, the temperature, relative humidity, oxygen and carbon dioxide concentration of the chamber was found within the acceptable limits. The particle size MMAD and GSD values were within the acceptable limits, at the mean limit concentration of 5.04 mg/L of air.

All animals were observed for clinical signs of toxicity and mortality during exposure and post exposure on day 1 and once daily thereafter for clinical signs and twice daily for mortality for 14 days for post exposure period. Individual animal body weight was recorded on day 1 (on the day of exposure) prior to the exposure and on day 2, 4, 8 and 15. All the rats were euthanized after 14 days post exposure by intraperitoneal administration of sodium thiopentone and they were subjected to gross necropsy.

No treatment related clinical signs of toxicity and mortalities were observed. Slight decrease in body weight was noted on Day 2 due to exposure. All animals showed increase in body weight on day 4, 8 and 15.

No treatment related gross pathological findings were observed at the mean limit concentration of 5.04 mg/L of air.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

> 5.04 mg/L air

Physical form:

inhalation: aerosol

Quality of whole database:

reliable without restriction

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

see Rationale and Justification for the Analogue Read-Across Approach - Nanoforms and Bulk Forms (Chapter 13)

Reason / purpose for cross-reference:

read-across source

Limit test:

yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

other: Content C.I.

Remarks on result:

other: no animal died within 14 d observation period

Mortality:

No mortality was observed

Clinical signs:

other: No treatment-related effects were observed

Gross pathology:

No treatment-related effects were observed

Other findings:

No erythema or oedema was observed. Scratches were observed in 1 of 5 male animals.

Table Absolute Body Weights in g and Body Weight Gain in %:

Dose: 2000 mg/kg body weight
Animal No. / Sex	g Day 1	g Day 8	g Day 15	% Day 1-15
21 / male	227	242	276	22
22 / male	236	251	277	17
23 / male	229	248	278	21
24 / male	230	241	274	19
25 / male	245	260	280	14
26 / female	216	220	223	3
27 / female	213	214	221	4
28 / female	218	215	217	-0.50
29 / female	215	218	223	4
30 / female	207	211	220	6

Table LD50:

Dose (Unit)	Number of Animals Investigated	Number of Intercurrent Deaths	LD50
2000 mg/kg bw	5 males	0	> 2000 mg/kg bw
2000mg/kg bw	5 females	0	> 2000 mg/kg bw

bw = body weight

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the present study, single dermal application of the test item to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity. In one male animal scratches were observed from day 6 until the end of the observation period which might be caused by scratching activities of the animal. The dermal LD50 was determined to be > 2000 mg / kg body weight.

Executive summary:

Acute toxicity after dermal application was investigated in Wistar rats according to an OECD 402 limit test. Animals (5 males and 5 females) were applied 2000 mg/kg bw in cottonseed oil for 24 hours. No animal died during the observation period.

Table1:  Results per Step

Sex	Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths
male	2000	5	0
female	2000	5	0

Signs of toxicity related to dose level used, time of onset and duration:

No treatment-related effects were observed

Effect on organs (related to dose level):

No treatment-related effects were observed.

Signs of irritation:

No erythema or oedema was observed. Scratches were observed in 1 of 5 male animals.

Conclusion

Under the conditions of the present study, single dermal application of the test item to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity. In one male animal scratches were observed from day 6 until the end of the observation period which might be caused by scratching activities of the animal.

The dermal LD50 was determined to be > 2000 mg / kg body weight.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Reliability of the original study (GLP compliant guideline study) as such was 1 (reliable without restrictions). Downgrading on reliability 2 is owed to the read across application. Due to their similar physicochemical properties it can reasonably be assumed that the target substance and the source substances used also reveal comparable toxicological properties and that the available toxicological data from the source substances can easily and reliably be used to predict specific properties of the target substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1114)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8
(drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept individually in IVC cages, type III H, polysulphone cages on Altromin
saw fibre bedding (lot no. 110811)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Type of coverage:

semiocclusive

Vehicle:

cotton seed oil

Details on dermal exposure:

Preparation of the Animals:
The animals were marked for individual identification by tail painting.
Approximately 24 hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper.
Care was taken to avoid abrading the skin, and only animals with healthy intact skin were used.
No less than 10% of the body surface was cleared for the application.
Prior to the application a detailed clinical observation was made of all animals.

Application:
The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface. In order to ensure good skin contact the test item was moistened with the vehicle.
The test item was held in contact with the skin by a dressing throughout a 24-hour period.
The dressing consisted of a gauze-dressing and non-irritating tape and was fixed with an additional dressing in a suitable manner.

Duration of exposure:

The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was removed using cottonseed oil.

Doses:

The test item was applied at a single dose of 2000 mg/kg body weight to each animal.

No. of animals per sex per dose:

5 male and 5 female

Control animals:

not required

Details on study design:

Observation period:
All animals were observed for 14 days after dosing

Weight Assessment:
The animals were weighed on day 1 (prior to the application) and on days 8 and 15.

Clinical Examination:
careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes
and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded.
Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities
were recorded. Cageside observations included changes in the skin and fur, eyes and mucous membranes.
Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour
pattern were examined. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea,
lethargy, sleep and coma.

Pathology:
At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected
intraperitoneally (Narcoren®, Merial) at the dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy.
All gross pathological changes were recorded and in case of findings the tissues were preserved for a possible
histopathological evaluation. The preserved tissues of which no histopathological evaluation was made will be discarded
3 months after the release of the final report unless otherwise agreed upon with the sponsor.

Evaluation of Results:
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by sex and dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

other: Content C.I.

Remarks on result:

other: no animal died within 14 d observation period

Mortality:

No mortality was observed

Clinical signs:

other: No treatment-related effects were observed

Gross pathology:

No treatment-related effects were observed

Other findings:

No erythema or oedema was observed. Scratches were observed in 1 of 5 male animals.

Table Absolute Body Weights in g and Body Weight Gain in %:

Dose: 2000 mg/kg body weight
Animal No. / Sex	g Day 1	g Day 8	g Day 15	% Day 1-15
21 / male	227	242	276	22
22 / male	236	251	277	17
23 / male	229	248	278	21
24 / male	230	241	274	19
25 / male	245	260	280	14
26 / female	216	220	223	3
27 / female	213	214	221	4
28 / female	218	215	217	-0.50
29 / female	215	218	223	4
30 / female	207	211	220	6

Table LD50:

Dose (Unit)	Number of Animals Investigated	Number of Intercurrent Deaths	LD50
2000 mg/kg bw	5 males	0	> 2000 mg/kg bw
2000mg/kg bw	5 females	0	> 2000 mg/kg bw

bw = body weight

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the present study, single dermal application of the test item to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity. In one male animal scratches were observed from day 6 until the end of the observation period which might be caused by scratching activities of the animal. The dermal LD50 was determined to be > 2000 mg / kg body weight.

Executive summary:

Acute toxicity after dermal application was investigated in Wistar rats according to an OECD 402 limit test. Animals (5 males and 5 females) were applied 2000 mg/kg bw in cottonseed oil for 24 hours. No animal died during the observation period.

Table1:  Results per Step

Sex	Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths
male	2000	5	0
female	2000	5	0

Signs of toxicity related to dose level used, time of onset and duration:

No treatment-related effects were observed

Effect on organs (related to dose level):

No treatment-related effects were observed.

Signs of irritation:

No erythema or oedema was observed. Scratches were observed in 1 of 5 male animals.

Conclusion

Under the conditions of the present study, single dermal application of the test item to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity. In one male animal scratches were observed from day 6 until the end of the observation period which might be caused by scratching activities of the animal.

The dermal LD50 was determined to be > 2000 mg / kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

adequate reliability

Justification for classification or non-classification

Available LD50 values of >10000 mg/kg bw (oral), >2000 mg/kg bw (dermal, RA from source substance PR 170) and the LC50 value of Y5.04 mg/L air exceed the respective threshold for classification. Thus the substance subject to registration does not meet criteria for classification according to REGULATION (EC) No 1272/2008.