Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex

Acute Toxicity: oral

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex

EC Number:

235-428-9

EC Name:

Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex

Cas Number:

12225-21-7

Molecular formula:

C48H33AlN12O27S6

IUPAC Name:

aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate)

Test material form:

solid: particulate/powder

Details on test material:

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.

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.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system 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

Administration / exposure

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

Results and discussion

Preliminary study:

not specified

Effect levels

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:

other: 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.

Gross pathology:

There were no gross pathological changes at necropsy.

Other findings:

not specified

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.