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

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2022

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

263-272-1

EC Name:

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

Cas Number:

61847-48-1

Molecular formula:

C33H24Cl2N4O6

IUPAC Name:

methyl 4-[(2,5-dichlorophenyl)carbamoyl]-2-({2-hydroxy-3-[(2-methoxyphenyl)carbamoyl]-1-naphthyl}diazenyl)benzoate

Test material form:

solid: nanoform, no surface treatment

Details on test material:

Name of test material (as cited in study report): Certificate of Analysis AZ653pc_specd1: Novoperm-Rot HF3S
Analytical purity: 95.3 % (w/w) (comp. 1)

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

Test animals

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.

Administration / exposure

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

Results and discussion

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)

Effect levels

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

Applicant's summary and conclusion

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.