Pyridinium, 1-[2-[[4-[[3-[2-[6-amino-1-hydroxy-3-sulfo-5-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-2-naphthalenyl]diazenyl]-4-sulfophenyl]amino]-4-oxobutyl]sulfonyl]ethyl]-3-carboxy-, inner salt, sodium salt (1:4)

Endpoint summary

Description of key information

Acute toxicity: via oral route

The LD50 of CR SB33 was greater than 2000 mg/kg B.W. (OECD TG423).

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:

From January 4, 2019 to September 16, 2019

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)

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: BioLASCO Taiwan Co., Ltd (Taipei, Taiwan)
- Age at study initiation: 8~12 week old
- Housing: two animals per cage
- Diet: ad libitum
- Water: ad libitum
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 15%
- Frequency of ventilation: 10~15 times/hour
- Photoperiod (hrs dark / hrs light): 12-hrs dark / 12-hrs light cycle

Route of administration:

oral: gavage

Vehicle:

water

Doses:

2000 mg/kg b.w.

No. of animals per sex per dose:

Three

Details on study design:

1. Test article was prepared to aqueous solution of 200 mg/mL in RO water. The dosage was 2,000 mg/kg BW.

2. After fasted overnight, the test article was administered in a single dose by gavage using a stomach tube.

3. First, three rats at group 1 were administered and observed for 48 hours. Group 2 were administered depending on the mortality results in the group 1.
3.1 If group 1 has <=1 out of three rats die, the same dose is administered to three rats in group 2 and their mortality rate is observed again.
3.2 If group 1 has >1 out of three rats die, a smaller dose is administered to three rats in group 2.

4. After the test article has been administered, food was withheld for a further 3 hours. Animals are observed individually after dosing at the 30 minutes and 4 hours, and continuous observation to day14.

5. Test item:
5.1 Record toxicity symptoms, time of their occurrence, duration, and rehabilitation details.
5.2 Mortality rate: The rate should be carefully observed and its condition should be recorded (once a day) for 14 days after the application of the test article.
5.3 Body weight: Individual weight of animals in day 1, in weekly intervals thereafter, and at the time of death or sacrifice.
5.4 Anatomical pathology and macropathology: A;; rats that survived the test should be checked for anatomical pathology and macropathology

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Based on:

test mat.

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

Group	Animal ID	Dosing Volumn (mL)	Body weight (g) Day1 Day7 Day14			Weight changes (g)
1	01F	1.9	182.4	207.0	217.1	+34.7
1	02F	2.0	196.3	233.2	249.6	+53.3
1	03F	2.1	207.1	244.1	268.2	+61.1
2	04F	2.1	202.2	239.7	252.7	+50.5
2	05F	2.0	192.0	233.9	233.7	+41.7
2	06F	1.9	187.4	219.3	229.4	+42.0

^aWeight change = body weight (day 14)-body weight (day 1).

Table 2. Clinical observation of the rats

Group

Animal ID

Clinical sign observation ^a

30 mins

4 hours

D2 ^b

D3

D4

D5

D6

D7

D8

D9

D10

D11

D12

D13

D14

1

01F

N ^c

N

N

N

N

N

N

N

N

N

N

N

N

N

N

1

02F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

1

03F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

2

04F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

2

05F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

2

06F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

^aClinical sign observation including: skin, hair, eyes, activity, circulatory system, respiratory system, digestive system and urinary system.

^bD X: Day X of test.

^cN: Normal.

Table 3. Results of gross necropsy examination

Group	Animal ID	Dose	Gross lesion
1	01F	2,000 mg/kg BW	No significant lesion founded
	02F		No significant lesion founded
	03F		No significant lesion founded
2	04F	2,000 mg/kg BW	No significant lesion founded
	05F		No significant lesion founded
	06F		No significant lesion founded

^aGross lesion including: appearance, subcutis, abdomen, reproductive system, spleenm pancreas, liver, stomach, intestines, mesemteric lymph node, kidney, urinary bladder, adrenal, thorax, thymus, heart, lung, oral cavity, salivary gland, lymph nodes, muscles, esophagus, trachea and other.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

According to OECD 423 test method, the LD50 of CR SB33 was greater than 2000 mg/kg B.W.. Therefore, CR SB33 was Category 5 based on GHS criteria.

Executive summary:

This test using the procedures outlined in the SuperLab for MZ6-181200026 which is based on the SOP (SOPP-341) for the OECD 423 and OECD 423 (OECD, 2001). A total of 6 female Sprague-Dawley rats were orally dosed with CR SB33 in three animals each group, at 2000 mg/kg b.w. for both Group 1 and Group 2 in limit test. 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 CR SB33 was greater than 2,000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Limited exposure evisaged.

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Limited exposure evisaged.

Additional information

Acute toxicity: via oral route

A total of 6 female Sprague-Dawley rats were orally dosed with CR SB33 in three animals each group, at 2000 mg/kg b.w. for both Group 1 and Group 2 in limit test. 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 CR SB33 was greater than 2,000 mg/kg.