Reaction products of Hydrogen amino-4-hydroxynaphthalene-2-sulfonate, Potassium substituted-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate and Sodium substituted-({4-[(2-chloroethyl)sulfonyl]butanoyl}amino)benzenesulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid

Details on test material:

Test item : FAT 40875/A TE
Physical appearance : Dark red powder
Purity as per CoA : 86.4% all organic components
Batch No. : BOP 05-17 (BS-ROE 1550 NIM 25+26+27)
Manufactured date : 20 December 2017
Expiry date : 11 December 2022
pH : 4.3 (aq. soln. 2 %w/w at room-temperature)
Recommended storage condition: Refrigeration (+2 to +8 °C) and protection from light

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Test item: FAT 40875/A TE
- Physical appearance: Dark red powder
- Source and lot/batch No.of test material: Huntsman Textile Effects (Germany) GmbH / BOP 05-17 (BS-ROE 1550 NIM 25+26+27)
- Expiration date of the lot/batch: 11 December 2022
- Purity test date: 86.4% all organic components

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (+2 to +8 °C), Protect from light.

OTHER SPECIFICS:
- measurement of pH: 4.3 (aq. soln.(2% (w/w)) at room-temperature.

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, No.93, Solur, Thally Road, Anekal, Bengaluru - 562106, India.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 Weeks
- Weight at study initiation: 180.7 to 217.6 g
- Fasting period before study: 16 to 18 hours
- Housing: Rats were housed individually in standard polysulfone cages (Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottle. Additionally, polycarbonate rat huts were placed inside the cage as an 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, manufactured by Krishna Valley Agro Tech LLP, MIDC Kupwad block, Sangli, Maharashtra, was provided to animals.
- 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 manufactured by Eureka Forbes Ltd, Mumbai 400001, India was provided to animals in polycarbonate bottles with stainless steel sipper tubes.
- Acclimation period: After physical examination for good health and suitability for experiment, the animals were acclimatized for six to fourteen days before treatment. Animals were observed once daily during acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 24
- Humidity (%): 65 to 67
- Air changes (per hr): 14.6
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Milli-Q water

Details on oral exposure:

The prepared dose formulation was administered once orally as gavage to the fasted (16 to 18 hours) rats. Each animal was administered orally by gavage using disposable plastic syringe attached with metal feeding canula. The dose volume was of 10 mL/kg body weight to deliver the dose of 300 mg/kg body weight (G1- FTS & STS) and 2000 mg/kg bodyweight (G2 -FTS & STS).

VEHICLE
- Concentration in vehicle: 30 mg/mL & 200 mg/mL
- Justification for choice of vehicle: The test item was found to be homogeneous and stable in the vehicle for 24 hours at the dose concentrations of 0.5 mg/mL and 250 mg/mL in the vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg.

Doses:

G1 - FTS : 300 mg/kg bw - Group 1, First test step.
G1 - STS : 300 mg/kg bw - Group 1, Second test step.
G2 - FTS : 2000 mg/kg bw - Group 2, First test step.
G2 - STS : 2000 mg/kg bw - Group 2, Second test step.

No. of animals per sex per dose:

Three per step

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.

- Frequency of observations and weighing: 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 to15 post administration. The body weights were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days 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 toxic signs were recorded.

- Necropsy of survivors performed: The rats surviving to the end of the observation period were euthanised by using isoflurane anaesthesia and subjected to detailed necropsy. Gross pathological findings were recorded and reported. Microscopic examination was not carried out as no gross pathological changes were observed.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality observed at 300 & 2000 mg/kg bw.

Clinical signs:

No clinical signs observed at 300 & 2000 mg/kg bw.

Body weight:

The body weights of all the rats increased throughout the observation period in both 300 mg/kg bw and 2000 mg/kg bw.

Gross pathology:

No gross pathological changes were observed.

Any other information on results incl. tables

Group and dose (mg/kg body weight)	Rat No.	Sex	Body weight (g)						Day of death (Time of death)	No. dead/ no. tested	Pre-terminal deaths (%)
			Initial (Day 1)	Day 8	Weight change (Day 8 – Initial)	Day 15	Weight change (Day 15 – Initial)	At death
G1 (FTS)300	Rm9883	F	192.6	199.7	7.1	209.1	16.5	NA	NA	0/3	0
	Rm9884	F	210.0	217.7	7.7	226.9	16.9	NA	NA
	Rm9885	F	181.8	190.5	8.7	201.4	19.6	NA	NA
G1 (STS)300	Rm9886	F	182.8	201.9	19.1	208.7	25.9	NA	NA	0/3	0
	Rm9887	F	195.0	208.6	13.6	217.3	22.3	NA	NA
	Rm9888	F	203.5	212.1	8.6	222.6	19.1	NA	NA
G2 (FTS)2000	Rm9889	F	212.0	222.7	10.7	232.8	20.8	NA	NA	0/3	0
	Rm9890	F	184.9	198.4	13.5	210.5	25.6	NA	NA
	Rm9891	F	200.6	211.0	10.4	219.2	18.6	NA	NA
G2(STS)2000	Rm9892	F	180.7	190.3	9.6	200.8	20.1	NA	NA	0/3	0
	Rm9893	F	209.9	220.7	10.8	227.4	17.5	NA	NA
	Rm9894	F	217.6	224.2	6.6	232.1	14.5	NA	NA

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for FAT 40875/A TE was found to be greater than 5000 mg/kg.

Executive summary:

The acute oral toxicity study of FAT 40875/A TE to the Wistar rats was evaluated according to OECD 423 test guideline.

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 (First treatment step) at a dose volume of 10 mL/kg body weight to attain the dose of 300 mg/kg body weight. No clinical signs were observed and there were no pre-terminal deaths.

Based on the scheme - Annex 2c, three additional female rats were tested at the same dose of 300 mg/kg body weight G1 - STS (Second treatement step). No clinical signs were observed and there were no pre-terminal deaths.

The treatment was continued with the next higher dose of 2000 mg/kg bodyweight – G2-FTS. No clinical signs were observed and there were no pre-terminal deaths. Three additional female rats were tested at the same dose of 2000 mg/kg body weight (G2-STS). No clinical signs were observed and there were no pre-terminal deaths.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing and again on days 8 and 15. Necropsy was performed for all the survived rats at terminal sacrifice. All survived rats gained weight during experimental period. There were no gross pathological findings at necropsy.

Based on the results of the present study, the LD₅₀for FAT 40875/A TE is greater than 5000 mg/kg as per the LD₅₀cut-off value.

The test item does not meet the classification criteria as per Globally Harmonized Classification system as there were no mortality or clinical signs of toxicity observed till 2000 mg/kg body weight.