Methyl 4-[[(2,5-dichlorophenyl)amino]carbonyl]-2-[[2-hydroxy-3-[[(2-methoxyphenyl)amino]carbonyl]-1-naphthyl]azo]benzoate

Endpoint summary

Description of key information

Oral toxicity

 

The test item (Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single oral gavage administration to male and female rats at 5000 mg/kg bw in a OECD guideline and GLP compliant study. The LD50 (male/female rat) was greater than 5000 mg/kg body weight.

 

Inhalation

The acute inhalation (4 hours) Median Lethal Concentration (LC50) value of “Novoperm-Rot HF3S” is more than 5.05 mg/L of chamber air in Wistar rats.

 

Dermal toxicity

The Test item (<90% Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single dermal application to male rats at 5000 mg/kg bw in a OECD guideline compliant study. The LD50 (male rat) was greater than 5000 mg/kg body weight.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

key study

Study period:

July 1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to OECD Guideline 401 and GLP

Justification for type of information:

See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 188 using Pigment Red 22, Pigment Red 112 and Pigment Orange 38 as Source Substances (Chapter 13)

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

EEC Directive 79-831 Annex V, Part B 4.1.1.

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:

TEST ANIMALS
- Strain specifics: Hoe: WISKf (SPF71)
- Source: Hoechst AG, Kastengrund, SPF-breed
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: male 191 g - 199 g, female 187 g - 208 g
- Fasting period before study: approximately 16 hours before treatment, access to water permitted
- Housing: in groups of five in Makrolon type 4 cages with standard softwood bedding
- Diet (e.g. ad libitum): standard rat diet (Albtromin 1324) ad libidum
- Water (e.g. ad libitum): tap water ad libidum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 10 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/ml
- Amount of vehicle (if gavage): 20 ml/kg body weight, application volume was distributed over one hour (test item in vehicle administered)

Doses:

5000 mg/kg body weight

No. of animals per sex per dose:

5 males
5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days starting with treatment day 1
- Frequency of observations and weighing:
mortality/viability: during the first 30 minutes and approximately 1, 2, and 4 h after administration on day 1 and daily on days 2-15
clinical signs: during the first 30 minutes and approximately 1, 2, 3 and 6 h after administration on day 1 and daily on days 2-15
body weights: on days 1 (prior to administration), 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macroscopic examination

Statistics:

None

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

no deaths

Clinical signs:

other: During the first hours reduced spontaneous activity, and after 2-4 hours hunched posture and diarrhea with stained feces were observed. No clinical signs were noted at 1 day after administration or later.

Gross pathology:

No macroscopic findings at scheduled necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

67/548/EEC: Acute oral toxicity: no classification warranted
1272/2008/EC: Acute oral toxicity: no classification warranted

The test item (Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single oral gavage administration to male and female rats at 5000 mg/kg bw in a OECD guideline and GLP compliant study. The LD50 (male/female rat) was greater than 5000 mg/kg body weight.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

key study

Study period:

July 1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to OECD Guideline 401 and GLP

Justification for type of information:

See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 188 using Pigment Red 22, Pigment Red 112 and Pigment Orange 38 as Source Substances (Chapter 13)

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

EEC Directive 79-831 Annex V, Part B 4.1.1.

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:

TEST ANIMALS
- Strain specifics: Hoe: WISKf (SPF71)
- Source: Hoechst AG, Kastengrund, SPF-breed
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: male 191 g - 199 g, female 187 g - 208 g
- Fasting period before study: approximately 16 hours before treatment, access to water permitted
- Housing: in groups of five in Makrolon type 4 cages with standard softwood bedding
- Diet (e.g. ad libitum): standard rat diet (Albtromin 1324) ad libidum
- Water (e.g. ad libitum): tap water ad libidum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 10 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/ml
- Amount of vehicle (if gavage): 20 ml/kg body weight, application volume was distributed over one hour (test item in vehicle administered)

Doses:

5000 mg/kg body weight

No. of animals per sex per dose:

5 males
5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days starting with treatment day 1
- Frequency of observations and weighing:
mortality/viability: during the first 30 minutes and approximately 1, 2, and 4 h after administration on day 1 and daily on days 2-15
clinical signs: during the first 30 minutes and approximately 1, 2, 3 and 6 h after administration on day 1 and daily on days 2-15
body weights: on days 1 (prior to administration), 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macroscopic examination

Statistics:

None

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

no deaths

Clinical signs:

other: During the first hours reduced spontaneous activity, and after 2-4 hours hunched posture and diarrhea with stained feces were observed. No clinical signs were noted at 1 day after administration or later.

Gross pathology:

No macroscopic findings at scheduled necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

67/548/EEC: Acute oral toxicity: no classification warranted
1272/2008/EC: Acute oral toxicity: no classification warranted

The test item (Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single oral gavage administration to male and female rats at 5000 mg/kg bw in a OECD guideline and GLP compliant study. The LD50 (male/female rat) was greater than 5000 mg/kg body weight.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to OECD Guideline 401 and GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

EEC Directive 79-831 Annex V, Part B 4.1.1.

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:

TEST ANIMALS
- Strain specifics: Hoe: WISKf (SPF71)
- Source: Hoechst AG, Kastengrund, SPF-breed
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: male 191 g - 199 g, female 187 g - 208 g
- Fasting period before study: approximately 16 hours before treatment, access to water permitted
- Housing: in groups of five in Makrolon type 4 cages with standard softwood bedding
- Diet (e.g. ad libitum): standard rat diet (Albtromin 1324) ad libidum
- Water (e.g. ad libitum): tap water ad libidum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 10 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/ml
- Amount of vehicle (if gavage): 20 ml/kg body weight, application volume was distributed over one hour (test item in vehicle administered)

Doses:

5000 mg/kg body weight

No. of animals per sex per dose:

5 males
5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days starting with treatment day 1
- Frequency of observations and weighing:
mortality/viability: during the first 30 minutes and approximately 1, 2, and 4 h after administration on day 1 and daily on days 2-15
clinical signs: during the first 30 minutes and approximately 1, 2, 3 and 6 h after administration on day 1 and daily on days 2-15
body weights: on days 1 (prior to administration), 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macroscopic examination

Statistics:

None

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

no deaths

Clinical signs:

other: During the first hours reduced spontaneous activity, and after 2-4 hours hunched posture and diarrhea with stained feces were observed. No clinical signs were noted at 1 day after administration or later.

Gross pathology:

No macroscopic findings at scheduled necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

67/548/EEC: Acute oral toxicity: no classification warranted
1272/2008/EC: Acute oral toxicity: no classification warranted

The test item (Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single oral gavage administration to male and female rats at 5000 mg/kg bw in a OECD guideline and GLP compliant study. The LD50 (male/female rat) was greater than 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

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

Adequacy of study:

key study

Study period:

2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study was performed using the bulk nano form of the registered substance.

Justification for type of information:

Please see read across justification document in chapter 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 433 (Acute Inhalation Toxicity: Fixed Concentration Procedure)

Principles of method if other than guideline:

The test item dust aerosol was generated and injected at a specific rate into the inhalation chamber. Groups of animals of a single male sex were exposed for 4-hours period of time to the test chemical in a stepwise procedure using the appropriate fixed concentrations for dusts/mists (aerosols) as set out in Annex 1. As the rats were observed ‘evident toxicity’ rather than death/moribundity was used as an endpoint. Concentrations that were expected to be lethal were avoided.
The initial concentration level was selected on the basis of a sighting study at the concentrations of a 1mg/l and 5mg/L and there were no clinical signs were observed in the rats at this sighting study exposure concentration.
The rats were observed for effects in terms of toxic signs and pre-terminal deaths. The surviving rats at the end of the study were necropsied

GLP compliance:

yes

Test type:

fixed concentration procedure

Limit test:

yes

Specific details on test material used for the study:

Test Item Information (As furnished by the Sponsor)

Batch Manufactured By
(Name and Address) : Clariant Produkte (Deutschland) GmbH
Brueningstrasse 50
65929 Frankfurt am Main,
Germany

Batch Supplied By
(Name and Address) : Clariant Produkte (Deutschland) GmbH
Brueningstrasse 50
65929 Frankfurt am Main,
Germany

Test Item : Novoperm-Rot HF3S

Color Index (as per Certificate of Analysis) : Pigment Red 188

CAS NO. : 61847-48-1

Chemical Name (IUPAC) : Methyl4-[[(2,5-dichlorophenyl)-amino]-carbonyl]-2-[[2-hydroxy-3-[[(2-methoxyphenyl)-amino]-carbonyl]-1-naphthyl]-azo]-benzoate

Physical Appearance : Red powder

Purity as Certificate of Analysis : 95.3 % (w/w)

Batch No. : DEB2136860

Manufactured date : 08.11.2010

Expiry date : 08.11.2022

Recommended Storage Condition : Ambient (+15 to + 25°C)

Physico-chemical properties : pH : 5 to 8
Density: 1.47 g/mL
Water solubility: <0.02 mg/L

Note: 1. Date of receipt of test item at test facility: 19 October 2021
2. Test item code by test facility: C067-06

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

Rats were housed individually per cage per sex for sighting study and for limit test in group of two / three per cage / sex in standard polysulfone cages (Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelletted food and drinking water. Additionally, polycarbonate rat huts were placed inside the cage as an enrichment object and was changed along with the cage once a week.
Bedding: Steam sterilized corn cob was used and changed along with the cage once a week.

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Mass median aerodynamic diameter (MMAD):

2.35 µm

Geometric standard deviation (GSD):

1.71

Remark on MMAD/GSD:

The value for Mass Median Aerodynamic Diameter (MMAD) 1-4 µm and geometric standard deviation (GSD) 1.5 to 3.0 for the test item were within the range for G1 group as recommended by OECD guidance document no. 39 on acute inhalation testing

Details on inhalation exposure:

The inhalation toxicity study of “Novoperm-Rot HF3S” was determined in 5 male Wistar rats by exposure to test item as a dust aerosol generated by dust generator with an air flow rate of 10 LPM and a piston speed of 92 mm/h with 200 kilopascals of atomizer pressure. The rats were housed in special rat restrainers and continuously exposed to the test item aerosol (nose only) for 4 hours in an inhalation exposure chamber (dynamic state) for the G1 group. The post treatment observation period was 14 days

Duration of exposure:

4 h

Concentrations:

The average actual concentration of “Novoperm-Rot HF3S” was 5.05 mg/L of chamber air. The test item concentration at the animals’ breathing zone was determined using gravimetric filter analysis method

No. of animals per sex per dose:

5

Details on study design:

Five male rats per group was assigned to main group for limit test, by body weight stratification during acclimatization. Animals with extreme body weights were not used for the study and excluded from the treatment.
Group allocation, piston speed and number of rats

Group Piston speed
(mm/h) Sex No. of
rats Rat Numbers
From To

G1
92
M

5

Rad3371

Rad3375

M: Male

Preliminary study:

8.2.5.1 Technical pre-test
The test item was made into fine powder using marshall mars mill and sieve.
A technical pre-test was conducted without animals to check the feasibility of generating adequate test atmosphere (Test item concentration, particle size distribution, O2, CO2, temperature and relative humidity) after chamber equilibrium. The chamber air actual concentration was estimated gravimetrically.

The exposure conditions and results of technical pre-test is given below at highest achieved actual test item concentration at breathing zone.

Technical pre-test exposure conditions:
Aerosol generation system Dust generator
Feeding air supply pressure 200 Kilopascals
Piston speed (mm/h) /Airflow rate (LPM) 92 / 10
Sampling duration 1 min
Sampling rate 4 LPM

Technical pre-test results:
Test item actual concentration
(mg/L of air) 5.02
MMAD (µm) & GSD 2.39 & 1.71
O2 content in % v/v 20.8
CO2 content in % v/v 0.03
Chamber temperature in °C 21.7
Relative humidity (%) 62.3

8.2.5.2 Sighting study
Sighting study I:
As no evidence from existing data on the same chemical and structurally related chemicals, hence as per Annex 1 the starting concentration 1 mg/L was selected for dust aerosol.

As per Annex 1 of fixed concentration procedure, the sighting study was conducted at 18 mm/h piston speed and at 10 LPM airflow rate with using dust generator as per OECD 433 traditional protocol using 1 male and female rat to achieve the 1 mg/L chamber air concentration.

Sighting study I exposure conditions:
Aerosol generation system Dust generator
Feeding air supply pressure 200 Kilopascals
Piston speed (mm/h) /Airflow rate (LPM) 18 / 10
Sampling duration 1 minute
Sampling rate (LPM) 4

The animals were exposed in inhalation chamber with dynamic airflow.

The results of sighting study I are given below.
Group No. of animals and sex 1) Test item concentration # in Chamber air, mg/L
2) MMAD (µm) & GSD
3)Nominal Concentration (mg/L) Clinical
signs*
G1 1 M
1 F 1)

Sex:

male

Dose descriptor:

LC50

Remarks:

Based on the results of the present study, the test item “Novoperm-Rot HF3S” is classified as follows: • The test item is classified as category 5 / unclassified as per Annex 1 of the OECD 433 test guideline furnished as Annexure 1 in the report. • The te

Effect level:

>= 5.05 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Clinical signs:

other: All animals were normal after the exposure period

Body weight:

All rats gained bodyweight throughout the experiment period

Gross pathology:

No abnormality was detected at necropsy in any of the exposed rats

Interpretation of results:

Category 5 based on GHS criteria

Remarks:

Based on the results of the present study, the test item is classified as follows: • The test item is classified as category 5 / unclassified as per Annex 1 of the OECD 433 test guideline furnished as Annexure 1 in the report. • The test item is classified as category 5 as per Globally Harmonized System of Classification and Labelling of Chemicals (GHS) Ninth Revised Edition, United Nations (2021). ST/SG/AC.10/30/Rev.9, as there was no mortality at 5.05 mg/L of chamber air in male Wistar rats (Appendix 3).

Conclusions:

The acute inhalation (4 hours) Median Lethal Concentration (LC50) value of “Novoperm-Rot HF3S” is more than 5.05 mg/L of chamber air in Wistar rats.

Executive summary:

An acute inhalation toxicity study with Pigment Red 188 was conducted in Wistar rats with one treatment group (G1).

The inhalation toxicity was determined in 5 male Wistar rats by exposure to test item as a dust aerosol generated by dust generator with an air flow rate of 10 LPM and a piston speed of 92 mm/h with 200 kilopascals of atomizer pressure. The rats were housed in special rat restrainers and continuously exposed to the test item aerosol (nose only) for 4 hours in an inhalation exposure chamber (dynamic state) for the G1 group. The post treatment observation period was 14 days.

The dust aerosol was sampled from the inhalation chamber to determine particle size distribution, Mass Median Aerodynamic Diameter (MMAD) and Geometric standard deviation (GSD). Particle size of the aerosol generated in the chamber air was determined two times i.e., during the first and fourth hour of the exposure period using any one port. The MMAD and GSD was calculated using validated excel spread sheet.

The overall mean MMAD of 2.35 micrometers and GSD of 1.71 was observed for G1 group. 

The average actual concentration of test item was 5.05 mg/L of chamber air. The test item concentration at the animals’ breathing zone was determined using gravimetric filter analysis method.

All animals were normal after the exposure period.

All rats gained bodyweight throughout the experiment period. There were no mortalities observed during the experimental period. No abnormality detected at necropsy.

The acute inhalation (4 hours) Median Lethal Concentration (LC50) value is more than 5.05 mg/L of chamber air in Wistar rats.

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2022

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 433 (Acute Inhalation Toxicity: Fixed Concentration Procedure)

Principles of method if other than guideline:

The test item dust aerosol was generated and injected at a specific rate into the inhalation chamber. Groups of animals of a single male sex were exposed for 4-hours period of time to the test chemical in a stepwise procedure using the appropriate fixed concentrations for dusts/mists (aerosols) as set out in Annex 1. As the rats were observed ‘evident toxicity’ rather than death/moribundity was used as an endpoint. Concentrations that were expected to be lethal were avoided.
The initial concentration level was selected on the basis of a sighting study at the concentrations of a 1mg/l and 5mg/L and there were no clinical signs were observed in the rats at this sighting study exposure concentration.
The rats were observed for effects in terms of toxic signs and pre-terminal deaths. The surviving rats at the end of the study were necropsied

GLP compliance:

yes

Test type:

fixed concentration procedure

Limit test:

yes

Specific details on test material used for the study:

Test Item Information (As furnished by the Sponsor)

Batch Manufactured By
(Name and Address) : Clariant Produkte (Deutschland) GmbH
Brueningstrasse 50
65929 Frankfurt am Main,
Germany

Batch Supplied By
(Name and Address) : Clariant Produkte (Deutschland) GmbH
Brueningstrasse 50
65929 Frankfurt am Main,
Germany

Test Item : Novoperm-Rot HF3S

Color Index (as per Certificate of Analysis) : Pigment Red 188

CAS NO. : 61847-48-1

Chemical Name (IUPAC) : Methyl4-[[(2,5-dichlorophenyl)-amino]-carbonyl]-2-[[2-hydroxy-3-[[(2-methoxyphenyl)-amino]-carbonyl]-1-naphthyl]-azo]-benzoate

Physical Appearance : Red powder

Purity as Certificate of Analysis : 95.3 % (w/w)

Batch No. : DEB2136860

Manufactured date : 08.11.2010

Expiry date : 08.11.2022

Recommended Storage Condition : Ambient (+15 to + 25°C)

Physico-chemical properties : pH : 5 to 8
Density: 1.47 g/mL
Water solubility: <0.02 mg/L

Note: 1. Date of receipt of test item at test facility: 19 October 2021
2. Test item code by test facility: C067-06

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

Rats were housed individually per cage per sex for sighting study and for limit test in group of two / three per cage / sex in standard polysulfone cages (Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelletted food and drinking water. Additionally, polycarbonate rat huts were placed inside the cage as an enrichment object and was changed along with the cage once a week.
Bedding: Steam sterilized corn cob was used and changed along with the cage once a week.

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Mass median aerodynamic diameter (MMAD):

2.35 µm

Geometric standard deviation (GSD):

1.71

Remark on MMAD/GSD:

The value for Mass Median Aerodynamic Diameter (MMAD) 1-4 µm and geometric standard deviation (GSD) 1.5 to 3.0 for the test item were within the range for G1 group as recommended by OECD guidance document no. 39 on acute inhalation testing

Details on inhalation exposure:

The inhalation toxicity study of “Novoperm-Rot HF3S” was determined in 5 male Wistar rats by exposure to test item as a dust aerosol generated by dust generator with an air flow rate of 10 LPM and a piston speed of 92 mm/h with 200 kilopascals of atomizer pressure. The rats were housed in special rat restrainers and continuously exposed to the test item aerosol (nose only) for 4 hours in an inhalation exposure chamber (dynamic state) for the G1 group. The post treatment observation period was 14 days

Duration of exposure:

4 h

Concentrations:

The average actual concentration of “Novoperm-Rot HF3S” was 5.05 mg/L of chamber air. The test item concentration at the animals’ breathing zone was determined using gravimetric filter analysis method

No. of animals per sex per dose:

5

Details on study design:

Five male rats per group was assigned to main group for limit test, by body weight stratification during acclimatization. Animals with extreme body weights were not used for the study and excluded from the treatment.
Group allocation, piston speed and number of rats

Group Piston speed
(mm/h) Sex No. of
rats Rat Numbers
From To

G1
92
M

5

Rad3371

Rad3375

M: Male

Preliminary study:

8.2.5.1 Technical pre-test
The test item was made into fine powder using marshall mars mill and sieve.
A technical pre-test was conducted without animals to check the feasibility of generating adequate test atmosphere (Test item concentration, particle size distribution, O2, CO2, temperature and relative humidity) after chamber equilibrium. The chamber air actual concentration was estimated gravimetrically.

The exposure conditions and results of technical pre-test is given below at highest achieved actual test item concentration at breathing zone.

Technical pre-test exposure conditions:
Aerosol generation system Dust generator
Feeding air supply pressure 200 Kilopascals
Piston speed (mm/h) /Airflow rate (LPM) 92 / 10
Sampling duration 1 min
Sampling rate 4 LPM

Technical pre-test results:
Test item actual concentration
(mg/L of air) 5.02
MMAD (µm) & GSD 2.39 & 1.71
O2 content in % v/v 20.8
CO2 content in % v/v 0.03
Chamber temperature in °C 21.7
Relative humidity (%) 62.3

8.2.5.2 Sighting study
Sighting study I:
As no evidence from existing data on the same chemical and structurally related chemicals, hence as per Annex 1 the starting concentration 1 mg/L was selected for dust aerosol.

As per Annex 1 of fixed concentration procedure, the sighting study was conducted at 18 mm/h piston speed and at 10 LPM airflow rate with using dust generator as per OECD 433 traditional protocol using 1 male and female rat to achieve the 1 mg/L chamber air concentration.

Sighting study I exposure conditions:
Aerosol generation system Dust generator
Feeding air supply pressure 200 Kilopascals
Piston speed (mm/h) /Airflow rate (LPM) 18 / 10
Sampling duration 1 minute
Sampling rate (LPM) 4

The animals were exposed in inhalation chamber with dynamic airflow.

The results of sighting study I are given below.
Group No. of animals and sex 1) Test item concentration # in Chamber air, mg/L
2) MMAD (µm) & GSD
3)Nominal Concentration (mg/L) Clinical
signs*
G1 1 M
1 F 1)

Sex:

male

Dose descriptor:

LC50

Remarks:

Based on the results of the present study, the test item “Novoperm-Rot HF3S” is classified as follows: • The test item is classified as category 5 / unclassified as per Annex 1 of the OECD 433 test guideline furnished as Annexure 1 in the report. • The te

Effect level:

>= 5.05 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Clinical signs:

other: All animals were normal after the exposure period

Body weight:

All rats gained bodyweight throughout the experiment period

Gross pathology:

No abnormality was detected at necropsy in any of the exposed rats

Interpretation of results:

Category 5 based on GHS criteria

Remarks:

Based on the results of the present study, the test item is classified as follows: • The test item is classified as category 5 / unclassified as per Annex 1 of the OECD 433 test guideline. • The test item is classified as category 5 as per Globally Harmonized System of Classification and Labelling of Chemicals (GHS) Ninth Revised Edition, United Nations (2021). ST/SG/AC.10/30/Rev.9, as there was no mortality at 5.05 mg/L of chamber air in male Wistar rats (Appendix 3).

Conclusions:

The acute inhalation (4 hours) Median Lethal Concentration (LC50) value of is more than 5.05 mg/L in Wistar rats.

Executive summary:

An acute inhalation toxicity study with Pigment Red 188 was conducted in Wistar rats with one treatment group (G1).

The inhalation toxicity was determined in 5 male Wistar rats by exposure to test item as a dust aerosol generated by dust generator with an air flow rate of 10 LPM and a piston speed of 92 mm/h with 200 kilopascals of atomizer pressure. The rats were housed in special rat restrainers and continuously exposed to the test item aerosol (nose only) for 4 hours in an inhalation exposure chamber (dynamic state) for the G1 group. The post treatment observation period was 14 days.

The dust aerosol was sampled from the inhalation chamber to determine particle size distribution, Mass Median Aerodynamic Diameter (MMAD) and Geometric standard deviation (GSD). Particle size of the aerosol generated in the chamber air was determined two times i.e., during the first and fourth hour of the exposure period using any one port. The MMAD and GSD was calculated using validated excel spread sheet.

The overall mean MMAD of 2.35 micrometers and GSD of 1.71 was observed for G1 group. 

The average actual concentration was 5.05 mg/L of chamber air. The test item concentration at the animals’ breathing zone was determined using gravimetric filter analysis method.

All animals were normal after the exposure period.

All rats gained bodyweight throughout the experiment period. There were no mortalities observed during the experimental period. No abnormality detected at necropsy.

The acute inhalation (4 hours) Median Lethal Concentration (LC50) value of is more than 5.05 mg/L of chamber air in Wistar rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC0

Value:

5 mg/L air

Physical form:

inhalation: dust

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:

experimental study

Adequacy of study:

key study

Study period:

11 October - 12 November 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The test was performed in accordance with OECD, but not according to GLP guidelines and the English test report was reconstructed by the test institute from the study raw data and the original Czech report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding Farm VELAZ, monitoring quality
- Age at study initiation: 7-9 weeks
- Weight at study initiation: 225-275 g
- Fasting period before study: not stated
- Housing: 5 animals per plastic breeding cage VELAZ T4
- Diet (e.g. ad libitum): ST 1 Bergman - standard pellet diet ad libitum, Mill Kocanda, Jesenice u Prahy, Czech Republic
- Water (e.g. ad libitum): dinking tap water ad libitum
- Acclimation period: minumum 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22° ± 3°C (air-condition)
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 4x6 cm (shaved 24 hours before application)
- Type of wrap if used: application site was covered by mull, plastic foil and held in contact by plaster (strapping)


REMOVAL OF TEST SUBSTANCE
- Washing (if done): after 24-hour application period, the remains of the test substance were removed from the skin
- Time after start of exposure: 24 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5000 mg/kg
- For solids, paste formed: yes


VEHICLE
- Amount(s) applied (volume or weight with unit): water was used to moisten test substance

Duration of exposure:

24 hours

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5 males

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical examination was done after 30 min, 3 hours of the 1st day, in the moring and afternoon of the second day and thereafter daily for 14 days; body weights were recorded before application, atday 8 and before euthanasia of animals
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight:

Statistics:

not relevant

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

All animals survived the 14-day observation period.

Clinical signs:

other: No clinical signs were observed during the 14-day observation period.

Gross pathology:

All animals were without gross morphological alterations.

Other findings:

none

Interpretation of results:

GHS criteria not met

Conclusions:

67/548/EEC: Acute oral toxicity: no classification warranted
1272/2008/EC: Acute oral toxicity: no classification warranted

The Test item (<90% Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single dermal application to male rats at 5000 mg/kg bw in a OECD guideline compliant study. The LD50 (male rat) was greater than 5000 mg/kg body weight.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Study period:

11 October - 12 November 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The test was performed in accordance with OECD, but not according to GLP guidelines and the English test report was reconstructed by the test institute from the study raw data and the original Czech report.

Justification for type of information:

See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 188 using Pigment Red 22, Pigment Red 112 and Pigment Orange 38 as Source Substances (Chapter 13)

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding Farm VELAZ, monitoring quality
- Age at study initiation: 7-9 weeks
- Weight at study initiation: 225-275 g
- Fasting period before study: not stated
- Housing: 5 animals per plastic breeding cage VELAZ T4
- Diet (e.g. ad libitum): ST 1 Bergman - standard pellet diet ad libitum, Mill Kocanda, Jesenice u Prahy, Czech Republic
- Water (e.g. ad libitum): dinking tap water ad libitum
- Acclimation period: minumum 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22° ± 3°C (air-condition)
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 4x6 cm (shaved 24 hours before application)
- Type of wrap if used: application site was covered by mull, plastic foil and held in contact by plaster (strapping)


REMOVAL OF TEST SUBSTANCE
- Washing (if done): after 24-hour application period, the remains of the test substance were removed from the skin
- Time after start of exposure: 24 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5000 mg/kg
- For solids, paste formed: yes


VEHICLE
- Amount(s) applied (volume or weight with unit): water was used to moisten test substance

Duration of exposure:

24 hours

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5 males

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical examination was done after 30 min, 3 hours of the 1st day, in the moring and afternoon of the second day and thereafter daily for 14 days; body weights were recorded before application, atday 8 and before euthanasia of animals
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight:

Statistics:

not relevant

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

All animals survived the 14-day observation period.

Clinical signs:

other: No clinical signs were observed during the 14-day observation period.

Gross pathology:

All animals were without gross morphological alterations.

Other findings:

none

Interpretation of results:

GHS criteria not met

Conclusions:

67/548/EEC: Acute oral toxicity: no classification warranted
1272/2008/EC: Acute oral toxicity: no classification warranted

The Test item (<90% Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single dermal application to male rats at 5000 mg/kg bw in a OECD guideline compliant study. The LD50 (male rat) was greater than 5000 mg/kg body weight.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Study period:

11 October - 12 November 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The test was performed in accordance with OECD, but not according to GLP guidelines and the English test report was reconstructed by the test institute from the study raw data and the original Czech report.

Justification for type of information:

See Rationale and Justification for the Analogue Read-Across Approach for the registration of the Nanoform of Pigment Red 188 using Pigment Red 22, Pigment Red 112 and Pigment Orange 38 as Source Substances (Chapter 13)

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding Farm VELAZ, monitoring quality
- Age at study initiation: 7-9 weeks
- Weight at study initiation: 225-275 g
- Fasting period before study: not stated
- Housing: 5 animals per plastic breeding cage VELAZ T4
- Diet (e.g. ad libitum): ST 1 Bergman - standard pellet diet ad libitum, Mill Kocanda, Jesenice u Prahy, Czech Republic
- Water (e.g. ad libitum): dinking tap water ad libitum
- Acclimation period: minumum 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22° ± 3°C (air-condition)
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 4x6 cm (shaved 24 hours before application)
- Type of wrap if used: application site was covered by mull, plastic foil and held in contact by plaster (strapping)


REMOVAL OF TEST SUBSTANCE
- Washing (if done): after 24-hour application period, the remains of the test substance were removed from the skin
- Time after start of exposure: 24 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5000 mg/kg
- For solids, paste formed: yes


VEHICLE
- Amount(s) applied (volume or weight with unit): water was used to moisten test substance

Duration of exposure:

24 hours

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5 males

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical examination was done after 30 min, 3 hours of the 1st day, in the moring and afternoon of the second day and thereafter daily for 14 days; body weights were recorded before application, atday 8 and before euthanasia of animals
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight:

Statistics:

not relevant

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

All animals survived the 14-day observation period.

Clinical signs:

other: No clinical signs were observed during the 14-day observation period.

Gross pathology:

All animals were without gross morphological alterations.

Other findings:

none

Interpretation of results:

GHS criteria not met

Conclusions:

67/548/EEC: Acute oral toxicity: no classification warranted
1272/2008/EC: Acute oral toxicity: no classification warranted

The Test item (<90% Pigment Red 112) did not cause any mortality or significant clinical signs or necropsy findings after single dermal application to male rats at 5000 mg/kg bw in a OECD guideline compliant study. The LD50 (male rat) was greater than 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 5 000 mg/kg bw

Justification for classification or non-classification

No classification, as no adverse effects were observed.