CJ303

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From October 29, 2019 to January 2, 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Preparation of test article: Stock solution of test article at concentration of 1,000.00 mg/L was prepared in reconstituted water. It was diluted in reconstituted water to desired concentrations for the test.
10mL of test solutions from control group and test group were sampled at the beginning (0 hour) and the end (96 hours) of the limit test, filtrated with 0.22µm filter membrane.

Vehicle:

yes

Remarks:

Reconstituted water

Details on test solutions:

The test article was stable, so the test was carried out using static system. The volume of test solution in each group was 500 mL.

Test organisms (species):

other: Zebrafish (Danio rerio)

Details on test organisms:

Test system:
1. Zebrafish (Danio rerio), the total body length was 1.0~2.0 cm.
2. Source: AZOO BIO CORPORATION(10F., No. 35, Ln. 3, Sec. 1, Zhongzheng E. Rd., Tamsui Dist., New Taipei City 251, Taiwan (R.O.C.))
3. Quality of fish: Within 7 days after the maintenance began, the mortality was less than 5%. As a result, the batch of fish was acceptable for the test.

Conditions:
1. Temperature: 23±2℃ during the maintenance. Light periodicity: 12~16 light cycle.
2. Feeding: one times at working day daily. Stop feeding 24 hours prior to the test.
3. Feed: Ayu Feed No.3 (Chuan Kuan, Kaohsiung Hsien, Taiwan).

Test type:

static

Water media type:

other: Reconstituted water

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

23±2℃

pH:

6.0 ~ 8.5

Dissolved oxygen:

Dissolved oxygen level of control group should be ≥60% saturated dissolved oxygen during the test.

Details on test conditions:

Light cycle: 16 hour: 8 hour (Light:Dark).
Feeding during the test and disturbances that may affect the behavior of the fish were avoided.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

9 375 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

1. Range-finding test:
1.1 The results showed that the numbers of death at 0.16, 0.80, 4.00, 20.00 and 100.00 mg/L nominal concentrations were 0, 0, 0, 0 and 0, respectively (Table 1). The mortality were 0.00%, 0.00%, 0.00%, 0.00% and 0.00%, respectively (Table 2).
1.2 The LC50 of test article was greater than the highest concentration group, so the limit test was conducted at 100.00 mg/L nominal concentration.

2. Limit test:
2.1 The results showed that the number of death at 100.00 mg/L nominal concentrations at the end of the test were 0 (Table 4). The mortality was 0.00% (Table 5).
2.2 The analytical results of test article concentration (Table 7) showed that the test article in control group was not detected at 0 and 96 hours. In test group, its concentration determined at 0 and 96 hours were 102.578 and 99.032 mg/L, respectively.

Sublethal observations / clinical signs:

Table 1. The number of fish died at each observation time for the range-finding test

Group	Nominal conc. (mg/L)	Number of death (individual)
		0 hour	2 hours	5 hours	24 hours	30 hours	48 hours	54 hours	72 hours	78 hours	96 hours
Control group	0.00	0	0	0	0	0	0	0	0	0	0
Test group	0.16	0	0	0	0	0	0	0	0	0	0
	0.80	0	0	0	0	0	0	0	0	0	0
	4.00	0	0	0	0	0	0	0	0	0	0
	20.00	0	0	0	0	0	0	0	0	0	0
	100.00	0	0	0	0	0	0	0	0	0	0

Table 2. The percentages mortality of fish at each observation time for the range finding test

Group	Nominal conc. (mg/L)	Mortality (%)
		0 hour	2 hours	5 hours	24 hours	30 hours	48 hours	54 hours	72 hours	78 hours	96 hours
Control group	0.00	0.00^a	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
Test group	0.16	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
	0.80	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
	4.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
	20.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
	100.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00

^aMortality = Dead number / Total number * 100%

Table 3. Light intensity, temperature, dissolved oxygen and pH value for the range-finding test

Group	Nominal conc. (mg/L)	Time (hour)	Light intensity (lux)	Temperature (℃)	Dissolved oxygen level (%)	pH
Control group	0.00	0	920	24.6	100.0	7.75
		24	930	25.0	86.3	7.30
		48	940	25.0	97.2	7.00
		72	920	24.9	94.2	6.99
		96	930	25.0	93.6	6.97
Test group	100.00	0	920	24.6	99.6	7.80
		24	930	25.0	85.8	7.33
		48	940	25.0	96.2	7.08
		72	920	24.9	84.8	7.06
		96	930	25.0	85.5	7.04

Table 4. The number of fish died at each observation time for the limit test

Group	Nominal conc. (mg/L)	Number of death (individual)
		0 hour	2 hours	5 hours	24 hours	30 hours	48 hours	54 hours	72 hours	78 hours	96 hours
Control group	0.00	0	0	0	0	0	0	0	0	0	0
Test group	100.00	0	0	0	0	0	0	0	0	0	0

Table 5. The percentages mortality of fish at each observation time for the limit test

Group	Nominal conc. (mg/L)	Number of death (individual)
		0 hour	2 hours	5 hours	24 hours	30 hours	48 hours	54 hours	72 hours	78 hours	96 hours
Control group	0.00	0.00^a	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
Test group	100.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00

^aMortality = Dead number / Total number * 100%

Table 6. Light intensity, temperature, dissolved oxygen and pH value for the limit test

Group	Nominal conc. (mg/L)	Time (hour)	Light intensity (lux)	Temperature (℃)	Dissolved oxygen level (%)	pH
Control group	0.00	0	920	25.0	100.0	7.40
		24	930	24.9	96.3	7.37
		48	920	25.0	92.1	7.32
		72	930	24.9	91.1	7.08
		96	920	24.9	89.4	7.06
Test group	100.00	0	920	25.0	100	7.33
		24	930	24.9	94.8	7.37
		48	920	25.0	91.7	7.35
		72	930	24.9	90.3	7.15
		96	920	24.9	87.3	7.09

Table 7. The results of concentration analysis for the limit test

Group	Nominal conc. (mg/L)	Sampling time (hour)^a	Analytial conc. (mg/L)
Control group	0	0	0
		96	0
Test group	100	0	102.578
		96	99.032

^aThe sampling dates (yyyy/mm/dd) of  0 hour and 96 hours were 2019/11/11 and 2019/11/15, repectively.

Validity criteria fulfilled:

yes

Conclusions:

This study was conducted according OECD203:2019 to evaluate the lethal effect of test article on zebrafish (Danio rerio). The results indicated that under the conditions designed for this test, CJ303 was not lethal to zebrafish. Its NOEC was 100.00 mg/L nominal concentration.

Executive summary:

This study was conducted according to OECD 203:2019 to evaluate fish acute toxicity caused by CJ303. The test system was zebrafish (Danio rerio). The nominal concentrations of test article in the range-finding test were 0.16~100.00 mg/L. The result showed that the LC50 was greater than the highest concentration of test groups, so the limit test was conducted at 100.00 mg/L nominal concentration. The results of the limit test showed that the nominal concentrations of 100.00 mg/L caused mortality of fish at the 96 hour was 0.00%. Therefore, under the conditions designed for this test, CJ303 was not lethal to zebrafish. Its no observed effect concentration (NOEC) was 100.00 mg/L nominal concentration.

Description of key information

The 96-hr LC50 of CJ303 was 9375 mg/L for fish (OECD TG203).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

9 375 mg/L

Additional information

This study was conducted according OECD203:2019 to evaluate the lethal effect of test article on zebrafish (Danio rerio). The results indicated that under the conditions designed for this test, CJ303 was not lethal to zebrafish. Its NOEC was 100.00 mg/L nominal concentration.