Registration Dossier

Administrative data

Description of key information

Acute Oral Toxicity: 

The acute oral toxicity dose (LD50) was considered based on experimental study conducted on rats for the test chemical. The LD50 value is >2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity: 

This data was considered for waiver considering the low vapour pressure of this chemical (0.000000000000000000000913 Pa) as well as the particle size distribution. Majority of the particles were found to be in the size 150 (62.5%) micrometer in size which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex is highly unlikely.

Acute Dermal Toxicity:

The acute dermal toxicity dose (LD50) was considered based on experimenatl study conducted on rats for the test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental study report.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The objective of this acute oral toxicity study was to assess the toxicological profile of the test item when administered to rats by a single oral gavage.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
Name of test material: Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex
- IUPAC name: aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate)
- Molecular formula: C48H33AlN12O27S6
- Molecular weight: 1429.19 g/mole
-Smiles:C1=CC(=CC=C1N=NC2C(=NN(C2=O)C3=CC=C(C=C3)S(=O)(=O)O)C(=O)O)S(=O)(=O)[O-].C1=CC(=CC=C1N=NC2C(=NN(C2=O)C3=CC=C(C=C3)S(=O)(=O)O)C(=O)O)S(=O)(=O)[O-].C1=CC(=CC=C1N=NC2C(=NN(C2=O)C3=CC=C(C=C3)S(=O)(=O)O)C(=O)O)S(=O)(=O)[O-].[Al+3]
- Inchl: 1S/3C16H12N4O9S2.Al/c3*21-15-13(18-17-9-1-5-11(6-2-9)30(24,25)26)14(16(22)23)19-20(15)10-3-7-12(8-4-10)31(27,28)29;/h3*1-8,13H,(H,22,23)(H,24,25,26)(H,27,28,29);/q;;;+3/p-3
- Substance type: Organic
- Physical appearance : Yellow to greenish-yellow powder.
- Purity as per Certificate of Analysis : 40.79%
- Lot No. : FG/16-17/0427
- Manufactured date : 19 May 2016
- Expiry Date : Shelf life of 8 years
- pH : 3.98 at 26.8°C
- Density : 0.283 g/cm3 at 26.8°C
- Storage conditions : Ambient (+15 to +25°C)
- SAFETY PRECAUTIONS: Gloves, cap and face mask were used in addition to protective body garments and shoes, to ensure adequate personal health and safety and to avoid inhalation and skin contact with the test item.
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Geniron Biolabs Pvt. Ltd. Bengaluru
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 to 12 weeks
- Weight at study initiation: 176.24 g to 215.59 g
- Identification:By rat accession number. Identification of individual rats was by cage card and turmeric colour body markings. The rat accession number was allotted during the course of the study. The temporary body marking during acclimatization period was done with crystal violet.
- Fasting period before study: rats were fasted for approximately 16 to 18 hours
- Housing:Rats were housed individually in standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill
- Diet (e.g. ad libitum): Hypro Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, Mumbai, ad libitum
- Acclimation period: The animals were acclimatized six days for G1-FTS, eight days for G1-STS, thirteen days for G2-FTS before treatment and and fifteen days for G2-STS.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 24°C
- Humidity (%): 65 to 67 %
- Air changes (per hr): air conditioned with adequate fresh air supply (between 13.1 and 13.2 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: 27 June 2018 To: 26 July 2018
Route of administration:
oral: gavage
Vehicle:
other: Milli-Q water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300 & 2000 mg/kg body weight.
- Amount of vehicle (if gavage):dose volume was 30 & 200 mg/ml body weight
Doses:
G1 (FTS) - 300 mg/kg
G1 (STS) - 300 mg/kg
G2 (FTS) - 2000 mg/kg
No. of animals per sex per dose:
G1 (FTS) - 300 mg/kg - 3
G1 (STS) - 300 mg/kg - 3
G2 (FTS) - 2000 mg/kg - 3
G2 (STS) - 2000 mg/kg - 3
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Clinical signs and pre-terminal deaths:At each step, the animals were observed five times on test day 1 (day of administration) i.e. at 30 minutes and four times at hourly intervals and once daily during days 2 to 15 post administration. Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to the observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma and all observed clinical signs were recorded.
- Body weights:The body weights were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).
- Necropsy of survivors performed: yes,The rats surviving to the end of the observation period were euthanised by using isoflurane anaesthesia and subjected to detailed necropsy.
- Other examinations performed:Gross pathological findings were recorded and reported. Microscopic examination was not carried out as no gross pathological changes were observed.
Statistics:
not specified
Preliminary study:
not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality waso bserved
Mortality:
no mortality was observed
Clinical signs:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: There were
no clinical sings and pre-terminal deaths.
G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: There
were no clinical signs observed in any of the rats.
Body weight:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: The body
weights of all the rats increased throughout the observation period.
G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: The body
weights of all the rats increased throughout the observation period.
Gross pathology:
There were no gross pathological changes at necropsy.
Other findings:
not specified

Table 1:Body weight, body weight change and pre-terminal deaths

 

Group/Step and Dose (mg/kg

body weight)

Rat No.

Sex

Body weight (g)

Day of Death(Time of

Death)

No. dead/

No.

tested

Preterminal deaths (%)

 

Initial

(Day 1)

8th day

Weight

change

(day 8 –

Initial)

15th day

 

Weight

change

(day 15

– Initial)

At Death

G1

(FTS)

300

Rw289

F

213.81

224.67

10.86

229.53

15.72

NA

NA

0/3

 

0

Rw290

F

196.42

204.31

7.89

209.71

13.29

NA

NA

Rw291

F

178.91

190.92

12.01

198.59

19.68

NA

NA

G1

(STS)

300

Rw292

F

209.56

217.13

7.57

225.20

15.64

NA

NA

0/3

0

Rw293

F

215.59

223.08

7.49

228.69

13.10

NA

NA

Rw294

F

212.47

220.57

8.10

227.07

14.60

NA

NA

G2

(FTS)

2000

Rw295

F

209.51

217.68

8.17

223.16

13.65

NA

NA

0/3

0

Rw296

F

214.36

221.92

7.56

229.43

15.07

NA

NA

Rw297

F

176.24

187.84

11.60

199.23

22.99

NA

NA

G2

(STS)

2000

Rw298

F

210.73

219.34

8.61

224.12

13.39

NA

NA

0/3

0

Rw299

F

176.75

188.41

11.66

200.01

23.26

NA

NA

Rw300

F

180.01

188.49

8.48

199.46

19.45

NA

NA

F: Female FTS: First Treatment Step STS: Second Treatment Step NA: Not Applicable 0: No deaths

Table 2: Individual clinical signs, dose administration and necropsy findings

Group/Step and

Dose

(mg/kg

body weight)

Date and

time of

administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume

administered

(mL)

Day of observation

Day 1

30 min

1hr

2hr

3hr

4hr

G1 (FTS)

300

03 July 2018 and

11:42 AM

To 11:45 AM

Rw289

F

213.81

2.14

N

N

N

N

N

Rw290

F

196.42

1.96

N

N

N

N

N

Rw291

F

178.91

1.79

N

N

N

N

N

 

Group/Step and

Dose

(mg/kg

body weight)

Rat

No.

 

Sex

Day of observation

Necropsy findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G1

(FTS)

300

Rw289

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw290

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw291

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female FTS: First treatment step N: Normal min: minutes mg: milligrams kg: kilograms mL: millilitre NAD: No Abnormality Detected

Table 2 contd. Individual clinical signs, dose administration and necropsy findings

Group/Step and

Dose

(mg/kg

body weight)

Date and

time of

administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume

administered

(mL)

Day of observation

Day 1

30 min

1hr

2hr

3hr

4hr

G1 (STS) 300

05 July 2018 and

12:04 PM

to

12:06 PM

Rw292

F

209.56

2.10

N

N

N

N

N

Rw293

F

215.59

2.16

N

N

N

N

N

Rw294

F

212.47

2.12

N

N

N

N

N

 

Group/Step and

Dose

(mg/kg

body weight)

Rat

No.

 

Sex

Day of observation

Necropsy findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G1

(STS)

300

Rw292

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw293

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw294

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

 F: Female STS: Second treatment step N: Normal min: minutes mg: milligrams kg: kilograms mL: millilitre NAD: No Abnormality Detected

Table 2 contd. Individual clinical signs, dose administration and necropsy findings

Group/Step and

Dose

(mg/kg

body weight)

Date and

time of

administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume

administered

(mL)

Day of observation

Day 1

30 min

1hr

2hr

3hr

4hr

G2

(FTS)

2000

10 July 2018 and

11:43 AM

to

11:45 AM

Rw295

F

209.51

2.10

N

N

N

N

N

Rw296

F

214.36

2.14

N

N

N

N

N

Rw297

F

176.24

1.76

N

N

N

N

N

 

Group/Step and

Dose

(mg/kg

body weight)

Rat

No.

 

Sex

Day of observation

Necropsy findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G2

(FTS)

2000

Rw295

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw296

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw297

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female FTS: First treatment step N: Normal min: minutes mg: milligrams kg: kilograms mL: millilitre NAD: No Abnormality Detected

Table 2 contd. Individual clinical signs, dose administration and necropsy findings

Group/Step and

Dose

(mg/kg

body weight)

Date and

time of

administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume

administered

(mL)

Day of observation

Day 1

30 min

1hr

2hr

3hr

4hr

G2

(STS)

2000

12 July 2018 and

11:47 AM

to

11:49 AM

Rw298

F

210.73

2.11

N

N

N

N

N

Rw299

F

176.75

1.77

N

N

N

N

N

Rw300

F

180.01

1.80

N

N

N

N

N

 

Group/Step and

Dose

(mg/kg

body weight)

Rat

No.

 

Sex

Day of observation

Necropsy findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G2

(STS)

2000

Rw298

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw299

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw300

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female STS: Second treatment step N: Normal min: minutes mg: milligrams kg: kilograms mL: millilitre NAD: No Abnormality Detected

Interpretation of results:
other: not classified
Conclusions:
Based on the results of the present study, The LD50 value for test chemical Neelicert Tartrazine Lake (CAS No.: 12225-21-7) is considered to be >2000 mg/kg. The test item is “not classified (> 2000)” as per the criteria of CLP.
Executive summary:

The acute oral toxicity study with Neelicert Tartrazine Lake (CAS No.: 12225-21-7) in Wistar rats was conducted to assess the toxicological profile of the

test item.The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats

(G1-FTS) at the dose of 300 mg/kg body weight. There were no clinical signs of toxicity and pre-terminal deaths observed. As all the rats survived at this step, the test was confirmed with three additional female animals with the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths observed at this step. Based on the guideline OECD 423, the test was repeated at the dose of 2000 mg/kg body weight (G2-FTS). There were no clinical signs of toxicity observed. Hence, the test was confirmed with three additional female rats at the same dose of 2000 mg/kg body weight (G2-STS) as per the guideline OECD 423. There were no clinical signs of toxicity and pre-terminal deaths observed at this confirmatory test. Hence, the further dosing was stopped.The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Gross necropsy was performed for all the rats at termination. The body weight growth of all rats was unaffected by the test item. There were no gross pathological changes at necropsy, hence histopathology was not performed. Based on the results of the present study, the LD50 of test item, Neelicert tartrazine lake (CAS No.: 12225-21-7), the LD50 is >2000 mg/kg body weight.Thus, it was concluded that the acute toxicity study of Neelicert Tartrazine Lake (CAS No.: 12225-21-7), when administered via oral route in Wistar rats falls into the “Category not classified (> 2000)” criteria of CLP.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from experimental study report

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
Justification for data waiving:
other:
Endpoint conclusion
Quality of whole database:
Waiver

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
The objective of this acute dermal toxicity study was to assess the toxicological profile of the test item on application as a single semi-occlusive dermal application to rats.
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
- Name of test material: Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex
- IUPAC name: aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate)
- Molecular formula: C48H33AlN12O27S6
- Molecular weight: 1429.19 g/mole
-Smiles:C1=CC(=CC=C1N=NC2C(=NN(C2=O)C3=CC=C(C=C3)S(=O)(=O)O)C(=O)O)S(=O)(=O)[O-].C1=CC(=CC=C1N=NC2C(=NN(C2=O)C3=CC=C(C=C3)S(=O)(=O)O)C(=O)O)S(=O)(=O)[O-].C1=CC(=CC=C1N=NC2C(=NN(C2=O)C3=CC=C(C=C3)S(=O)(=O)O)C(=O)O)S(=O)(=O)[O-].[Al+3]
- Inchl: 1S/3C16H12N4O9S2.Al/c3*21-15-13(18-17-9-1-5-11(6-2-9)30(24,25)26)14(16(22)23)19-20(15)10-3-7-12(8-4-10)31(27,28)29;/h3*1-8,13H,(H,22,23)(H,24,25,26)(H,27,28,29);/q;;;+3/p-3- - - - - Substance type: Organic
- Physical state: Physical appearance : Yellow to greenish-yellow powder.
- Purity as per Certificate of Analysis: 40.79%
- Lot No. : FG/16-17/0427
- Manufactured date : 19 May 2016
- Expiry Date : Shelf life of 8 years
- pH : 3.98 at 26.8°C
- Density : 0.283 g/cm3 at 26.8°C
- Storage conditions : Ambient (+15 to +25°C)
- SAFETY PRECAUTIONS:Gloves, cap and face mask were used in addition to protective body garments and shoes, to ensure adequate personal health and safety and to avoid inhalation and skin contact with the test item.
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Geniron Biolabs Pvt. Ltd. Bengaluru
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 to 12 weeks
- Weight at study initiation: Females: 237.62 to 247.31 g
- Identification:By rat accession number. Identification of individual rats is by cage card and crystal violet colour body markings. The temporary body marking during acclimatization period was done with crystal violet. The rat accession numbers were allotted during the course of the study and was included in raw data and reported.
- Housing: Animals were housed individually in standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill. Additionally, polycarbonate rat huts were placed inside the cage as enrichment objects and were changed along with the cage once a week. Bedding: Steam sterilized corn cob was used and changed once a week along with the cage.
- Diet (e.g. ad libitum): Hypro Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, ad libitum
- Acclimation period: The rats were acclimatized for six, eight, twelve and fourteen days before treatment for dose range finding and main study respectively under standard laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 24°C
- Humidity (%): 65 to 67 %
- Air changes (per hr): air conditioned with adequate fresh air supply (between 13.1 and 13.2 air changes/hour)
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 27 June 2018 To: 26 July 2018
Type of coverage:
semiocclusive
Vehicle:
other: Milli-Q water
Details on dermal exposure:
TEST SITE
- Area of exposure: clipped skin of dorsolateral thoracic surface of the skin was clipped (approximately 10 x 8 cm)
- % coverage: 10% of the body surface
- Type of wrap if used: The applied area was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the applied area was washed with deionized water and wiped dry using clean towel.
- Time after start of exposure:24 hours
Duration of exposure:
24 hours
Doses:
DRF G1 - 200 mg/kg
DRF G2 - 1000 mg/kg
DRF G3 - 2000 mg/kg
Main G3 - 2000 mg/kg
No. of animals per sex per dose:
DRF G1 - 200 mg/kg - 1
DRF G2 - 1000 mg/kg - 1
DRF G3 - 2000 mg/kg - 1
Main G3 - 2000 mg/kg - 2
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Clinical examination and pre-terminal deaths:The animals were observed for clinical signs and pre-terminal deaths (mortality) once during first 30 minutes after application, and at hourly intervals for 6 hours after application on the day of treatment (day 1) and once daily during Days 2 to 15. In addition, treatment site was observed at 24, 48 and 72 hours after removal of test chemical using the Draize criteria (Refer Annexure 4 of this report). However, daily observation was done and animals did not exhibit any clinical signs. All rats were observed for changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Body weights - Individual body weights of animals were recorded on test days 1 (Pre-application), 8 (7 days post application), and 15 (14 days post application).
- Necropsy of survivors performed: yes,at the end of the observation period, all rats were euthanised and exsanguinated under isoflurane anesthesia and subjected to detailed necropsy by an experienced prosector and the findings were recorded.
- Other examinations performed: Microscopic examination was not carried out as no gross pathological changes were observed.
Statistics:
not specified
Preliminary study:
not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: not specified
Mortality:
There were no pre-terminal deaths (mortality) observed during the study.
Clinical signs:
There were no clinical signs observed during the study. There were no skin reactions at the site of application at 24 and 48 hours after test patch removal (as per draize method).
Body weight:
All rats gained body weight throughout the observation period.
Gross pathology:
No abnormality was detected at necropsy.
Other findings:
not specified

Table 1: Individual body weight, body weight changes and pre-terminal deaths

Group and Dose

(mg/kg body weight)

Rat No.

Sex

Body weight (g)

Pre-terminal deaths

Initial deaths

(Day 1 - at

treatment)

8thday

Weight change

(day 8 – Initial)

15thday

Weight change

(day 15 – Initial)

G1- DRF and 200

Rw301

F

237.62

244.19

6.57

249.38

11.76

0

G2- DRF and

1000

Rw302

F

247.31

251.02

3.71

257.27

9.96

0

G3 and 2000 DRF

Rw303

F

242.60

246.83

4.23

253.48

10.88

0

G3- Main study and

2000

Rw304

F

246.47

250.74

4.27

254.71

8.24

0

Rw305

F

241.21

248.03

6.82

256.09

14.88

0

DRF: Dose Range Finding    F: Female

Table 2: Individual test item application, clinical signs, skin reaction and necropsy findings

Dose range finding study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

 

Days

 

1

2

3

4

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

G1- DRF and

200

03 July 2018

and

11.47 AM

Rw301

F

237.62

48

N

N

N

N

N

N

N

N

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation/days

Necropsy findings

5

 

*

Er

@

Ed

@

6

7

8

9

10

11

12

13

14

15

 

G1- DRF and

200

Rw301

F

N

0

0

N

N

N

N

N

N

N

N

N

N

NAD

 

 

Table 2 contd. Individual test item application, clinical signs, skin reaction and necropsy findings

Dose range finding study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

 

Days

 

1

2

3

4

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

G2- DRF

and

1000

05 July 2018

and

12.06 PM

Rw302

F

247.31

247

N

N

N

N

N

N

N

N

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation/days

Necropsy findings

5

 

*

Er

@

Ed

@

6

7

8

9

10

11

12

13

14

15

 

G1- DRF

and

1000

Rw302

F

N

0

0

N

N

N

N

N

N

N

N

N

N

NAD

 

F: Female N: Normal h: hour min: minutes NAD: No abnormality detected Er: Erythema Ed: Edema Score 0: No Erythema / Edema

*: Clinical signs; @: Skin scoring as per Draize method (approximately 24, 48 and 72 hours) after test patch removal

 

Table 2 contd. Individual test item application, clinical signs, skin reaction and necropsy findings

Dose range finding study

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

 

Days

 

1

2

3

4

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

G3 and

2000

DRF

10 July 2018

and

11.47 AM

Rw303

F

242.60

485

N

N

N

N

N

N

N

N

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation/days

Necropsy findings

5

 

*

Er

@

Ed

@

6

7

8

9

10

11

12

13

14

15

 

G1 and

2000

DRF

Rw303

F

N

0

0

N

N

N

N

N

N

N

N

N

N

NAD

F: Female N: Normal h: hour min: minutes NAD: No abnormality detected Er: Erythema Ed: Edema Score 0: No Erythema / Edema

*: Clinical signs; @: Skin scoring as per Draize method (approximately 24, 48 and 72 hours) after test patch removal

Table 2 contd. Individual test item application, clinical signs, skin reaction and necropsy findings

Main study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

 

Days

 

1

2

3

4

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

G3and

2000

Main

12 July 2018

and

11.51 AM to

11.52 AM

Rw304

F

246.47

493

N

N

N

N

N

N

N

N

N

0

0

N

0

0

Rw305

F

241.21

482

N

N

N

N

N

N

N

N

N

0

0

N

0

0

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation/days

Necropsy findings

5

 

*

Er

@

Ed

@

6

7

8

9

10

11

12

13

14

15

 

G3 and

2000

Main

Rw304

F

N

0

0

N

N

N

N

N

N

N

N

N

N

NAD

Rw305

F

N

0

0

N

N

N

N

N

N

N

N

N

N

NAD

 

F: Female N: Normal h: hour min: minutes NAD: No abnormality detected Er: Erythema Ed: Edema Score 0: No Erythema / Edema

*: Clinical signs; @: Skin scoring as per Draize method (approximately 24, 48 and 72 hours) after test patch removal

Interpretation of results:
other: Not classified
Conclusions:
Based on the present study results, the acute dermal LD50 value of target chemical Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex(CAS No12225-21-7) is >2000 mg/kg body weight in female Wistar rats. The test item is classified as "Not classified”.
Executive summary:

The acute dermal toxicity of Neelicert tartrazine lake (CAS No.: 12225-21-7) in Wistar rats was tested with 200 (G1-DRF), 1000 (G2-DRF) and 2000 (G3-DRF) mg/kg with 1 female each for the dose range finding study and 2 female for main study (G3).Based on the individual body weight, the test item at the dose of 200, 1000 and 2000 mg/kg body weight was weighed on an aluminum foil and made as a paste in Milli-Q water and applied directly to the clipped skin of the animal to cover about 10% of the body surface of the animal(semi-occlusive). The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was for 24 hours.After the 24 hours contact period, the dressing was removed, and the applied area was washed with deionized water and wiped dry using clean towels.All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed at 24, 48 and 72 hours after removal of test item using the Draize criteria.There were no clinical signs of toxicity and mortality. There was no skin reaction observed at the site of application of test item. Body weights were measured on Days 1, 8 and 15 and growth was unaffected by test item. At the end of observation period (Day 15), all animals were euthanized and subjected to necropsy, and there were no gross lesions noted at the necropsy. Based on the present study results, the acute dermal LD50 of Neelicert tartrazine lake (CAS No.: 12225-21-7) is greater than 2000 mg/kg body weight in female Wistar rats.Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical Neelicert tartrazine lake (CAS No.: 12225-21-7) does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from experimental study report

Additional information

Acute Oral Toxicity: 

In the experimental study report, aluminium tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (12225-21-7) has been investigated for acute oral toxicity. The studies are summarized as below –

The acute oral toxicity study with Neelicert Tartrazine Lake (CAS No.: 12225-21-7) in Wistar rats was conducted to assess the toxicological profile of the

test item.The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats

(G1-FTS) at the dose of 300 mg/kg body weight. There were no clinical signs of toxicity and pre-terminal deaths observed. As all the rats survived at this step, the test was confirmed with three additional female animals with the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths observed at this step. Based on the guideline OECD 423, the test was repeated at the dose of 2000 mg/kg body weight (G2-FTS). There were no clinical signs of toxicity observed. Hence, the test was confirmed with three additional female rats at the same dose of 2000 mg/kg body weight (G2-STS) as per the guideline OECD 423. There were no clinical signs of toxicity and pre-terminal deaths observed at this confirmatory test. Hence, the further dosing was stopped.The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Gross necropsy was performed for all the rats at termination. The body weight growth of all rats was unaffected by the test item. There were no gross pathological changes at necropsy, hence histopathology was not performed. Based on the results of the present study, the LD50 of test item, Neelicert tartrazine lake (CAS No.: 12225-21-7), the LD50 is >2000 mg/kg body weight.Thus, it was concluded that the acute toxicity study of Neelicert Tartrazine Lake (CAS No.: 12225-21-7), when administered via oral route in Wistar rats falls into the “Category not classified (> 2000)” criteria of CLP.

Thus, based on the above experimental study on aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (122251-21-7) , it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (122251-21-7) cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity: 

This data was considered for waiver considering the low vapour pressure of this chemical (0.000000000000000000000913 Pa) as well as the particle size distribution. Majority of the particles were found to be in the size 150 (62.5%) micrometer in size which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex is highly unlikely.

Acute Dermal Toxicity:

In the experimental study report, aluminium tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (12225-21-7) has been investigated for acute dermal toxicity. The studies are summarized as below –

The acute dermal toxicity of Neelicert tartrazine lake (CAS No.: 12225-21-7) in Wistar rats was tested with 200 (G1-DRF), 1000 (G2-DRF) and 2000 (G3-DRF) mg/kg with 1 female each for the dose range finding study and 2 female for main study (G3).Based on the individual body weight, the test item at the dose of 200, 1000 and 2000 mg/kg body weight was weighed on an aluminum foil and made as a paste in Milli-Q water and applied directly to the clipped skin of the animal to cover about 10% of the body surface of the animal(semi-occlusive). The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was for 24 hours.After the 24 hours contact period, the dressing was removed, and the applied area was washed with deionized water and wiped dry using clean towels.All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed at 24, 48 and 72 hours after removal of test item using the Draize criteria.There were no clinical signs of toxicity and mortality. There was no skin reaction observed at the site of application of test item. Body weights were measured on Days 1, 8 and 15 and growth was unaffected by test item. At the end of observation period (Day 15), all animals were euthanized and subjected to necropsy, and there were no gross lesions noted at the necropsy. Based on the present study results, the acute dermal LD50 of Neelicert tartrazine lake (CAS No.: 12225-21-7) is greater than 2000 mg/kg body weight in female Wistar rats.Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical Neelicert tartrazine lake (CAS No.: 12225-21-7) does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Thus, based on the above experimental study on aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (122251-21-7) , it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (122251-21-7) cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies on aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (122251-21-7) and it’s structurally similar read across substances, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) (122251-21-7) cannot be classified for acute oral toxicity and acute dermal toxicity. For Acute inhalation toxicity wavier was added so, not possible to classify.