[No public or meaningful name is available]

Endpoint summary

Description of key information

Acute Oral Toxicity: The method used, was not intended for calculation of a precise LD50 value.

The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423. The test substance was classed as Category 5 in accordance with OECD Guideline No. 423.

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

Study period:

October 15 2020 to October 31 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

yes

Specific details on test material used for the study:

Not specified.

Species:

rat

Strain:

other:

Remarks:

Han:WIST

Sex:

female

Details on test animals or test system and environmental conditions:

Species and strain: Han:WIST rats
Source: TOXI COOP ZRT. Cserkesz u. 90. 1103 Budapest, Hungary
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: The Wistar rats as a rodent is one of the standard species of acute toxicity studies
Number of animals: 3 animals/group
Sex: Female, nulliparous and non pregnant animals
Age of animals: Young adult rat, 8 weeks old in first and second step
Body weight range at starting (first step): 167 - 170 g
Body weight range at starting (second step): 161 - 163 g
Acclimatization time: 7 days in first step and 8 days in second step

Husbandry
Animal health: Only healthy animals were used for the study. Health status was certified by the study director.
Room: 13/1
Housing: Group caging (3 animals/cage)
Cage type: Type III polypropylene/polycarbonate
Bedding: Certified laboratory bedding
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: above 10 air exchanges/hour by central air-condition system.

The temperature and relative humidity parameters were recorded daily during the study.

Food and Water Supply
Animals received ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany and tap water from municipal supply, as for human consumption from bottle ad libitum.
The diet and drinking water are periodically analysed and are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.

Identification
The individual identification was performed by numbers on the tail written by a permanent marker. The numbers were given on the basis of Toxi-Coop Zrt.'s master file, for each animal allocated to the study.
The boxes were identified by cards, holding information about study number, test item, sex, strain, dose group, date of arriving of animals, room number, cage number and individual animal numbers.

Route of administration:

oral: gavage

Vehicle:

other:

Remarks:

Sunflower oil, refined

Details on oral exposure:

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animals died in first step, so further three female rats were treated with the same dose. No animals died in second step, too, so the test was finished, because the stopping criteria of Annex 2d of OECD Guideline No. 423 was met.
Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried on the 15th day after the treatment.

Doses:

Justification of the doses
The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animals died in first step, so further three female rats were treated with the same dose. No animals died in second step, too, so the test was finished, because the stopping criteria of Annex 2d of OECD Guideline No. 423

No. of animals per sex per dose:

3 female rats per dose

Control animals:

not specified

Details on study design:

Procedure
A single oral administration - followed by a fourteen-day observation period - was performed by gavage. The day before treatment the animals were fasted. The food but not water was withheld overnight. Animals were weighed before the application and the food was given back 3 hours after the treatment.

Duration of the experimental period
7 days in first step and 8 days in second step of acclimatization, treatment’s day, 14 days post-treatment observation period, necropsy on Day 15.

Statistics:

Not specified.

Preliminary study:

Not applicable.

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other:

Remarks:

No lethality was noted at single oral dose of 2000 mg/kg bw.

Mortality:

No death occurred at 2000 mg/kg bw single oral dose of Dye-2019. All female rats in step 1 and step 2 survived until the end of the 14-day observation period.

Clinical signs:

In group 1 treated with 2000 mg/kg bw dose dark faeces (3 cases of 57 observations) were detected on Day 1. This alteration was observed in all animals (score +4).
In group 2 treated with 2000 mg/kg bw dose dark faeces (3 cases of 57 observations) were detected on Day 1. This alteration was observed in all animals (score +4).

Body weight:

The mean body weight of animals treated with 2000 mg/kg bw dose corresponded to their species and age throughout the study.

Gross pathology:

All animals treated with 2000 mg/kg bw dose survived until the scheduled necropsy on Day 15.
Greyish-greenish coloured kidneys was observed in two animals (No.: 9134, 9130).
No pathological changes were found related to the effect of the test item during the macroscopic examination of animals treated with 2000 mg/kg bw dose.

No death occurred after the single 2000 mg/kg bw oral dose of Dye-2019.  

There were no toxic clinical signs. The observed alteration as dark faeces was connected with the physical property of test item.

The test item caused a macroscopic alteration as greyish-greenish colouration in the kidneys, which was connected with the physical property of test item, probably.

There were no any changes related to the systemic toxic effect of the test item found in body weights and body weight gains during the study.

Autopsy revealed no treatment related pathological changes.

Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as below:

Dose (mg/kg bw)	Mortality (dead/treated)	LD50cut-off (mg/kg bw)	GHS category
2000	0/6 to 2/6	Between 2000 and 5000 Above 5000	5 5 or unclassified
300	0/6 to 2/6	Between 300 and 2000	4
50	0/6 to 2/6	Between 50 and 300	3
5	0/6 to 2/6	Between 5 and 50	2
5	3/6 or greater	Below 5	1

Summary of lethality – Post-treatment observation period (14 days

Groups	Treatment		Lethality
	Test item	Dose (mg/kg bw)	Females
1	Dye-2019 Step 1	2000	0/3
2	Dye-2019 Step 2	2000	0/3

 

Summary of clinical symptoms

Groups	Treatment		Symptoms	Incidence
	Test item	Dose mg/kg bw
1	Dye-2019 Step 1	2000	Dark faeces	3/57
			Normal	54/57
2	Dye-2019 Step 2	2000	Dark faeces	3/57
			Normal	54/57

Remark: Incidence = Number of symptoms/Summarized number of observations inside the group

Summarized number of observations inside the group =

(number of observations of first animal) + (number of observations of second animal) + (number of observations of third animal)

Summary of body weight gains (g)

Females	Day 0	Day 7	Day 15
Group 1: Dye-2019
2000 mg/kg bw; Step 1
Group size:	3	3	3
Mean (g)	168.0	197.7	207.3
SD:	1.73	2.08	1.53

 

Females	Day 0	Day 7	Day 15
Group 1: Dye-2019
2000 mg/kg bw; Step 1
Group size:	3	3	3
Mean (g)	162.0	194.3	203.7
SD:	1.00	3.51	7.02

 

Females	Day 0-7	Day 7-15	Day 0-15
Group 1: Dye-2019
2000 mg/kg bw; Step 1
Group size:	3	3	3
Mean (g)	29.7	9.7	39.3
SD:	3.06	1.15	3.06

Females	Day 0-7	Day 7-15	Day 0-15
Group 2: Dye-2019
2000 mg/kg bw; Step 2
Group size:	3	3	3
Mean (g)	32.3	9.3	41.7
SD:	4.51	3.51	8.02

Summarised gross pathology findings

Observations Females	Dye-2019 2000 mg/kg bw
Kidneys: Greyish-greenish colour	2/6
Kidneys: Normal	4/6

Interpretation of results:

GHS criteria not met

Conclusions:

The method used, was not intended for calculation of a precise LD50 value.
The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423. The test substance was classed as Category 5 in accordance with OECD Guideline No. 423.

Executive summary:

The objective of the acute oral study was to assess the toxicity of test item Dye-2019 when administered orally in a single dose to rats at one or more defined dose levels. The results of the study allowed the test item to be ranked according to most classification systems, currently in use. The study was carried out in accordance with OECD test guidline No.423 in compliance with GLP.

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animals died in first step, so further three female rats were treated with the same dose. No animals died in second step, too, so the test was finished, because the stopping criteria of Annex 2d of OECD Guideline No. 423 was met.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried on the 15th day after the treatment.

No lethality was noted at single oral dose of 2000 mg/kg bw.  

In first step, dark faeces were observed in all animals on Day 1. It was connected with the physical property of test item. No systemic clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal.  

In second step, dark faeces were observed in all animals on Day 1. It was connected with the physical property of test item. No systemic clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal.  

The body weight development was undisturbed in all animals.

The observed macroscopic alteration as greyish-greenish coloured kidneys was connected with the physical property of test item, probably. All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

The method used is not intended to allow the calculation of a precise LD50 value.

The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw