Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt

Endpoint summary

Description of key information

Acute toxicity: via oral route

The LD50 of Everzol Blue ED-G(Everzol SB26) was greater than 5000 mg/kg B.W. (EPA OPPTS 870.1100 and OECD TG401).

 

Acute toxicity: via dermal route

The LD50 of Everzol SB26 was greater than 2000 mg/kg B.W. (OECD TG402).

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:

Single dose

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: BioLASCO Taiwan Co., Ltd.
- Age at study initiation: about 5 week old
- Fasting period before study: overnight
- Housing: in polycarbonate cage
- Water: ad libitum
- Temperature (°C): 20-25 °C
- Humidity (%): 40-70 %
- Photoperiod: 12-hrs dark / 12-hrs light cycle

Route of administration:

oral: gavage

Vehicle:

water

Doses:

5000 mg/kg body weight

No. of animals per sex per dose:

For male: five
For female: five

Control animals:

yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

other: No adverse effects

Table 1. Symptoms and time course of death of rats administered by gavage with Everzol Blue ED-G

Sex	Dose (mg/kg)	Treated No.	Symptom	No. Observed											Total No. of death
				Hour			Day
				1	2	4	1	2	3	4	5	6	7	14
Male	0	5	Normal	5	5	5	5	5	5	5	5	5	5	5
			Death	0	0	0	0	0	0	0	0	0	0	0	0
	5,000	5	Normal	5	5	5	5	5	5	5	5	5	5	5
			Death	0	0	0	0	0	0	0	0	0	0	0	0
Female	0	5	Normal	5	5	5	5	5	5	5	5	5	5	5
			Death	0	0	0	0	0	0	0	0	0	0	0	0
	5,000	5	Normal	5	5	5	5	5	5	5	5	5	5	5
			Death	0	0	0	0	0	0	0	0	0	0	0	0

Table 2. Mortality rate of rats administered by gavage with Everzol Blue ED-G

Dose (mg/kg body weight)	Dead No./Treated No.			Mortality^1) (%)
	Male	Female	Total No.
Control	0/5	0/5	0/10	0
5,000	0/5	0/5	0/10	0

¹⁾ Mortality (%) = Dead No.÷Treated No. × 100%

Table 3. The body weight changes of rats administered by gavage with Everzol Blue ED-G on days 0, 7 and 14

Sex	Dose (mg/kg body weight)	Body weight (g)^1)
		Day 0	Day 7	Day 14
Male	Control	181.5 ± 4.7	205.2 ± 11.5	229.9 ± 20.8
	5,000	180.6 ± 4.2	219.1 ± 19.2	240.9 ± 31.3
Female	Control	159.3 ± 5.2	174.3 ± 4.5	198.4 ± 6.0
	5,000	159.2 ± 3.2	187.3 ± 11.8	203.1 ± 18.5

¹⁾ Mean ± S.D., n=5. Means in each column followed by asterisk (*) indicate significant difference at ρ<0.05 level by Student’st-test.

 

Table 4. The body weight gains of rats administered by gavage with Everzol Blue ED-G on days 7 and 14

Sex	Dose (mg/kg body weight)	Body weight gain (g)^1)
		Day 7^2)	Day 14^3)
Male	Control	23.6 ± 9.6	48.4 ± 18.7
	5,000	38.5 ± 15.7	60.3 ± 28.2
Female	Control	15.1 ± 3.6	39.1 ± 7.0
	5,000	28.1 ± 12.6	43.9 ± 20.0

¹⁾ Mean ± S.D., n=5. Means in each column followed by asterisk (*) indicate significant difference at ρ<0.05 level by Student’st-test.

²⁾ Body weight (BW) gain on the 7^thday = (7^th-0^th) BW (g).

³⁾ Body weight (BW) gain on the 14^thday = (14^th-0^th) BW (g).

Interpretation of results:

GHS criteria not met

Conclusions:

According to EPA OPPTS 870.1100 and OECD 401 test method, the LD50 of Everzol Blue ED-G was greater than 5000 mg/kg B.W.. Therefore, Everzol Blue ED-G is not classified according to CLP regulation.

Executive summary:

This test using the procedures outlined in the TACTRI for 0136Y07AORe which is based on the USEPA OPPTS 870.1100(US EPA 712-C-98-190) and OECD 401 (OECD, 2002). Wistar rats were administered by gavage with Everzol Blue ED-G at a fixed dose of 5,000 mg/kg body weight. At the end of experiment, all animals tolerated the test article well with increasing body weights and no mortality or gross lesions findings reported. The acute oral LD50 of Everzol Blue ED-G was greater than 5,000 mg/kg body weight in rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Endpoint conclusion:

no study available

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

Study period:

From August 22, 2016 to December 08, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: BioLASCO Taiwan Co. Ltd.
- Age at study initiation: about 7 weeks old
- Housing: Male and female rats were fed, respectively. Two rats per cage in an autoclaved polyethylene cage.
- Acclimation period: 7 Days
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 15%
- Photoperiod: 12 hrs dark / 12 hrs light

Type of coverage:

occlusive

Duration of exposure:

24 hours

Doses:

2000 mg/kg B.W.

No. of animals per sex per dose:

Six of male and six female

Control animals:

yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Table 1. Body weight of the rats in the study period

Group	Animal I.D.	Dosing volume (mL)	Body weight (g)		Weight chenges(g)
			Day 1	Day 14
Control	01M	0.5	246.6	345.4	+98.8
	02M	0.5	240.2	349.6	+109.4
	03M	0.5	266.4	355.2	+88.8
	04M	0.5	256.3	352.8	+96.5
	05M	0.6	283.5	385.8	+102.3
	06M	0.5	271.6	370.0	+98.4
Test	07M	0.5	267.7	359.7	+92.0
	08M	0.5	262.7	365.6	+102.9
	09M	0.5	249.4	356.9	+107.5
	10M	0.5	271.4	352.5	+81.1
	11M	0.5	264.1	361.1	+97.0
	12M	0.6	275.0	383.0	+108.0
Control	13F	0.4	204.6	260.2	+55.6
	14F	0.4	219.4	258.0	+38.6
	15F	0.4	220.5	275.2	+54.7
	16F	0.4	218.0	275.9	+57.9
	17F	0.5	230.2	270.9	+40.7
	18F	0.4	219.5	270.5	+51.0
Test	19F	0.4	206.0	254.0	+48.0
	20F	0.4	211.4	248.1	+36.7
	21F	0.5	225.1	287.3	+62.2
	22F	0.4	223.3	271.5	+48.2
	23F	0.4	224.7	282.6	+57.9
	24F	0.5	239.5	299.7	+60.2

Table 2. Clinical observation of the rats

Animal I.D.

Clinical sign observation

30 mins

4 hours

D2

D3

D4

D5

D6

D7

D8

D9

D10

D11

D12

D13

D14

01M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

02M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

03M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

04M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

05M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

06M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

07M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

08M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

09M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

10M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

11M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

12M

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

13F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

14F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

15F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

16F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

17F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

18F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

19F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

20F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

21F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

22F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

23F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

24F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

DX: Day X in the study period

N: Normal

Table 3. Results of gross necropsy examination

Animal I.D.	Dose	Gross lesion
01M	─	No significant lesion founded
02M		No significant lesion founded
03M		No significant lesion founded
04M		No significant lesion founded
05M		No significant lesion founded
06M		No significant lesion founded
07M	2000 mg/kg B.W.	No significant lesion founded
08M		No significant lesion founded
09M		No significant lesion founded
10M		No significant lesion founded
11M		No significant lesion founded
12M		No significant lesion founded
13F	─	No significant lesion founded
14F		No significant lesion founded
15F		No significant lesion founded
16F		No significant lesion founded
17F		No significant lesion founded
18F		No significant lesion founded
19F	2000 mg/kg B.W.	No significant lesion founded
20F		No significant lesion founded
21F		No significant lesion founded
22F		No significant lesion founded
23F		No significant lesion founded
24F		No significant lesion founded

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 402 test method, the LD50 of Everzol SB26 was greater than 2000 mg/kg B.W.. Therefore, Everzol SB26 was Category 5 based on GHS criteria.

Executive summary:

This test using the procedures outlined in the SuperLab for M62-151100129001EN which is based on the SOP (SOPP-342) for the OECD 402and OECD 402 (OECD, 1987). Six male and six femaleSprague-Dawley rats for each group were used in limit test. For Test group, 12Sprague-Dawley ratsweredermally dosed with 2000 mg/kg B.W. of Everzol SB26. All animals tolerated the test article well with increasing body weights and no mortality or gross lesions findings reported. In absence of mortality or other significant clinical signs of toxicity, LD50 ofEverzol SB26was greater than 2,000 mg/kg B.W..

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute toxicity: via oral route

Wistar rats were administered by gavage with Everzol Blue ED-G(Everzol SB26) at a fixed dose of 5,000 mg/kg body weight. At the end of experiment, all animals tolerated the test article well with increasing body weights and no mortality or gross lesions findings reported. The acute oral LD50 of Everzol Blue ED-G (Everzol SB26) was greater than 5,000 mg/kg body weight in rats.

 

Acute toxicity: via dermal route

Six male and six femaleSprague-Dawley rats for each group were used in limit test. For Test group, 12 Sprague-Dawley rats were dermally dosed with 2000 mg/kg B.W. of Everzol SB26. All animals tolerated the test article well with increasing body weights and no mortality or gross lesions findings reported. In absence of mortality or other significant clinical signs of toxicity, LD50 of Everzol SB26was greater than 2,000 mg/kg B.W..