2-[[4-[ethyl(2-hydroxyethyl)amino]phenyl]azo]-6-methoxy-3-methylbenzothiazolium methyl sulphate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

September 7-11, 1987

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: DIN 38 412 L15

Version / remarks:

Testverfahren mit Wasserorganismen (Gruppe L). Allgemeine Hinweise zur Planung, Durchführung und Auswertung biologischer Testverfahren (L1)" und "Bestimmung der Wirkung von Wasserinhaltsstoffen auf Fische - Fischtest (L15)", June 1982, using a static procedure

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final dilution of a dissolved solid, stock liquid or gel: In the form of an aqueous solution (10 g/100 mL) the product was added to the test water, subsequently the fish were placed into the aquaria

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

The test item was added to the test solution in form of an aqueous solution (10 g/100 ml). Subsequently the fish were places into the aquaria.

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Common name: Golden orfe
- Strain: Leuciscus idus L., golden variety
- Source: Fischzucht Paul Eggers, Hohenwestedt, Germany
- Length at study initiation (length definition, mean, range and SD): 5.1 cm (Range: 4.8 - 5.7)
- Weight at study initiation (mean and range, SD): 1.8 g (Range: 1.4 - 2.7)

ACCLIMATION
- Acclimation period: The duration of housing and adaptation was about 6 weeks, adaptation to test water and test temperature: 3 days
- Type and amount of food during acclimation: Withdrawal of food: 1 day
- Health during acclimation (any mortality observed): Mortality during the last 2 weeks of housing: about 0.1 %, mortality during the adaptation period: 0 %

FEEDING DURING TEST
- Food type: DRAFT: Growing feed F/B 50, SSNIFF Spezialdiäten GmbH, Soest, Germany
- Amount: ad lib.
- Frequency: ad lib.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

about 2.5 mM

Test temperature:

20 °C +/- 1 °C

pH:

about 7.9

Dissolved oxygen:

> 60 % of maximum saturation

Salinity:

not applicable

Nominal and measured concentrations:

Nominal: 0 (control), 10.0, 21.5, 46.4, 100.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel / housing: all-Glass aquarium (30 cm x 22 cm x 24 cm)
- Material, size, headspace, fill volume: glass aquarium, volume of water: 10 L
- Aeration: slight, continuous aeration with oil-free air
- Type of flow-through (e.g. peristaltic or proportional diluter): /
- Renewal rate of test solution (frequency/flow rate): /
- No. of animals per concentration: 10
- No. of vessels per control (replicates): 1
- Biomass loading rate: 1.8 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted freshwater according to DIN 38 412, Part 11, Draft Sept. 1981, preparation from fully demineralized tap water (conductivity max. 10 micro MHO), the resalting was carried out by addition of
* 294.0 mg/L CaCl2 x 2 H2O
* 123.3 mg/L MgSO4 x 7 H2O
* 64.8 mg/L NaHCO3
* 5.8 mg/L KCl
- Alkalinity: Acid capacity about 0.8 mM
- Ca/Mg ratio: 4:1


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light and 8 h darkness
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): tumbling, narcotic-like state

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 2.2
- Range finding study: Based on the results of a range-finding study (LC 50 after 96h: between 10 and 100 mg/L) the concentrations, spaced by a factor of about 2.2, were fixed as follows: 10.0, 21.5, 46.4 and 100.0 mg/L.
- Test concentrations: 10.0, 21.5, 46.4 and 100.0 mg/L.
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Chloroacetamide

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 46.4 - < 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 68.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: geometric mean value of LC0 and LC100

Results with reference substance (positive control):

LC50 (48 h) = 32 mg/L (determined on August 10, 1987)
This lethal concetration was stated to correspond to the normal sensitivity.

Reported statistics and error estimates:

Determination or calculation of the median lethal concentration (LC 50) and, if possible, the LC 5 and the LC 95 using the Probit analysis* was performed after 1, 4, 24, 48 72, and 96 h.

* modified according to FINNEY, D.J., PROBIT ANALYSIS, CAMBR. UNIV. PRESS, 3rd ED., 1971.

Sublethal observations / clinical signs:

Table 1: Results of mortality and observed symptoms during the test until the end (96 h). Since the test solutions were intensely colored a complete evaluation of the symptoms could not be performed.

Nominal conc. (mg/L)	Number of fish	Dead fish after
		1 h	4 h	24 h	48 h	72 h	96 h
10.0	10	0	0	0	0	0	0
21.5	10	0	0	0	0	0	0
46.4	10	0	0	0	0	0	0
100.0	10	0	0	0	2	7	10
0	10	0	0	0	0	0	0
Nominal conc. (mg/L)	Number of fish	Symptoms
		1 h	4 h	24 h	48 h	72 h	96 h
10.0	10
21.5	10
46.4	10
100.0	10				NT	NT
0.0
Explanation of Symptoms
A = Apathy	B = Abdominal Distension	E = Exophthalmos	F = Escape Reflex	H = Hyperreflexia	K = Convulsions
L = Gasping	N = Narcotic-Like state	T = Tumbling	U = Restlessness	V = Discoloration	W = Headstand	X = Accelerated respiration

Validity criteria fulfilled:

yes

Conclusions:

No observable effect concentration: 46.4 mg/L
Maximum concentration causing no mortality: 46.4 mg/L
Minimum concentration causing 100 % mortality: 100 mg/L

Executive summary:

In a 96-h acute toxicity study, Golden orfe (Leudiscus idus L.) were exposed to the test item at nominal concentrations of 0, 10.0, 21.5, 46.4, and 100.0 mg/L under flow through conditions.  The 96-h NOEC was 46.4 mg/L.  The 96-h LC₁₀₀ was 100.0 mg/L.  The 96-h LC₅₀ calculated from the geometric mean value of the LC₀ and LC₁₀₀ is 68.1 mg/L.

Sublethal effects of narcotic-like state and tumbling were observed in the groups exposed to 100.0 mg/L of the test material starting after 48-h of exposure. 

Based on the results of this study, the test item would be classified as Chronic 3 to L. idus in accordance with the CLP classification system.

This toxicity study is classified as acceptable and satisfies the guideline requirement for DIN 38 412 L15 toxicity study with L. idus.

 

Results Synopsis

 

Test organism size: Length 5.1 cm (Range: 4.8 - 5.7) and weight 1.8 g (Range: 1.4 - 2.7)

Test Type: Flowthrough

 

LC₅₀:  < 100 and > 46.4 mg/L

NOEC: 46.4 mg/L            

Endpoint(s) Effected: mortality

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

23.07.1980

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

not reported

GLP compliance:

no

Analytical monitoring:

no

Details on test solutions:

no data

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

no data

Test type:

not specified

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

no data

Test temperature:

16°C

pH:

6.9 - 8.1

Dissolved oxygen:

8.5-7.9

Salinity:

not applicable

Conductivity:

no data

Nominal and measured concentrations:

nominal: 10, 30, 100, 300 mg/L

Details on test conditions:

8 fish per concentration tested

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

14.2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Table 1: Results from the Acute toxicity Test with Oncorhynchus mykiss.

Concentration in mg/L	Surviving fish after X- hours
	4	24	48	72	96	pH	O2 in mg/L
10	8	8	8	8	7	7.9
30	8	8	6	5	4	7.9
100	8	8	4	3	3	8.1	8.5
300	0	0	0	0	0	6.9	7.9

Validity criteria fulfilled:

not specified

Conclusions:

The 96-h LC50 of the test item determined with rainbow trout is 50 mg/L.

Executive summary:

In a 96-h acute toxicity study according to OECD 203, rainbow trout (Oncorhynchus mykiss) were exposed to the test item at nominal concentrations of 10, 30, 100 and 300 mg/L. The active ingredient content in the test item was 28.5 %. The 96-h LC₅₀ was 50 mg/L. Also in the lowest concentration of 10.0 mg/L effects were observed, hence, the NOEC must be lower than 10.0 mg/L. Sublethal effects were not documented.

Based on the results of this study, a 100 % active ingredient (calculated LC₅₀ of 14.2 mg/L) would be classified as Chronic 3 to O. mykiss in accordance with the CLP classification system.

 

Results Synopsis

 

Test organism size: not reported

Test Type: not reported

 

96 h LC₅₀:  50.0 mg/L           

 

Endpoint(s) Effected: mortality

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

26.09.1983 - 29.09.1983

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

no data

GLP compliance:

no

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Test type:

not specified

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

25 °C

pH:

7.6 - 8

Dissolved oxygen:

7.8 - 8.7 mg/L

Nominal and measured concentrations:

Nominal: 10, 30, 100, 300 mg/L

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

17.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: geometric mean value of LC0 and LC100

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: remark: calculated from geometric mean LC50 of the test material

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

30 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Table 1: Results of surviving fish and corresponding medium parameter measured.

Concentration (in mg/L)	Number of living fish after X- hours
	1	4	24	48	72	96	pH	O2
10.0	10	10	10	10	10	10	7.6	8
30.0	10	10	10	8	6	0	7.8	7.8
100.0	10	10	0				8	8.1
300.0	10	0					7.6	8.7

Validity criteria fulfilled:

not specified

Conclusions:

The 96-h LC50 of the test item to Danio rerio is 22 mg/L.

Executive summary:

In a 96-h acute toxicity study, zebrafish (Danio rerio) were exposed to the test item (14.5 % active ingredient) at nominal concentrations of 10, 30, 100, and 300 mg/L. The 96-h NOEC was 10 mg/L. The 96-h LC₅₀ was 17.3 mg/L (geometric mean value of LC₀and LC₁₀₀).

Sublethal effects were not documented.  Based on the results of this study, the LC₅₀ for 100 % test item would be calculated as 2.5 mg/L, leading to a classification as classified as Chronic 2 to D. rerio in accordance with the CLP classification system.

 

Results Synopsis

Test organism size: not reported

Test Type: not reported

 

96 h LC₅₀:  22.0 mg/L (14.5 % act. ingr.)

96 h LC₀: 10 mg/L (14.5 % act. ingr.)           

 

Endpoint(s) Effected: mortality

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

06.11. - 10.11.1989

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

no data

GLP compliance:

no

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

23 °C

pH:

8.4 - 8.5

Dissolved oxygen:

90 - 94 %

Nominal and measured concentrations:

nominal: 3, 10, 30, 100 mg/L

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

8.78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

not specified

Conclusions:

The 96-h LC50 of the test item to Danio rerio is 8.78 mg/L.

Executive summary:

In a 96-h acute toxicity study, zebrafish (Danio rerio) were exposed to the test item at nominal concentrations of 3.0, 10, 30 and 100 mg/L. The 96-h NOEC was 3.0 mg/L. The 96-h LC₅₀ was 8.78 mg/L. Sublethal effects were not documented. Based on the results of this study the test item would be classified as Chronic 2 to D. rerio in accordance with the CLP classification system.

 

Results Synopsis

Test organism size: not reported

Test Type: not reported

 

96 h LC₅₀:  8.78 mg/L       

 

Endpoint(s) Effected: mortality

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

January 11- May 17, 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1984

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

not specified

Vehicle:

no

Details on test solutions:

For the test concentration 10 mg/I, a stock solution of 1 gil was prepared. For all other test concentrations, the stock solution was l0 gll. The test concentrations were achieved by addition of the appropriate volume of stock solution to the clilution water.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Upwey Trout Farm, Weymouth, Dorset, UK
- Length at study initiation: 51.5 mm - 52.0 mm
- Weight at study initiation: 1.33 g - 1.47g

ACCLIMATION
- Acclimation period: 16.04. - 19.04.1993; 23.04. - 26.04.1993; 30.04. - 04.05.1993
- Acclimation conditions (same as test or not): Temperature: 13-16 °C, Dissolved oxygen: > 80 % air saturation value; same as test
- Type and amount of food during acclimation: Trout Fry f'eed manufactured by BP Nutrition (U.K.) Limited.
- Health during acclimation (any mortality observed): The rainbow trout are examined prior to selection lbr the test to ensure that only healthy animals are exposed.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

240 mg/L CaCO3/L

Test temperature:

14.0 - 16.4 °C

pH:

7.6 - 8.7

Dissolved oxygen:

71 mg/L - 98 mg/L

Nominal and measured concentrations:

nominal: 10, 18, 32, 56, 100, 180 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Plastic aquaria with a working volume of 10 litre
- Material, size, headspace, fill volume: Tank containing 10 L of test solution
- Aeration: Continously diffusing air through the aquarium
- Type of flow-through (e.g. peristaltic or proportional diluter): The test solutions are replaced every 24 hours throughout the test period, the remaining live animals being transferred to an aquarium containing freshly prepared test solution.
- Renewal rate of test solution (frequency/flow rate): every 24 h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): /

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated mains water of typical hardness 240 mg calcium carbonate per litre
- Culture medium different from test medium: no
- Intervals of water quality measurement: before and after the daily change

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16:8

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: yes, but no results given

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 42 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 24 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

9.6 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: not documented
- Mortality of control: 0
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

48 h LC50 = 42 mg/L (95 % CI: 35-52)
72 h LC50 = 27 mg/L (95 % CI: 20-37)
96 h LC50 = 24 mg/L (95 % CI: 19-30)
LC50 are estimated graphically and 95% Confidence limits calculated where possible, according to the method of GRanmo and Larsstuvold: Evaluation of results from short-term tests. The LC50 value and its confidence limits. Food and Agriculture Organisation of the United Nations TF-INT 1976. 237 p.

Sublethal observations / clinical signs:

During the test the time of survival was recorded and is presented in table 1. An overview of the LC₅₀ values for different observation times and the corresponding confidence limits is presented in table 2.

Table 1.       Toxicity of Yoracryl Blue RGLN Liq 100 to Rainbow Trout Median Periods of Survival

Concentration  (mg/L)	Median Period of Survival in minutes with 95% confidence limits  ( - )
180	922      ( 784 - 1084)
100	2167      (1954 - 2403)
56	1451      (976 - 2157)
32	4740      (3976 - 5651)
18	20 % mortality in 96 h
10	No mortality
0	No mortality

 

Table 2.          Toxicity of Yoracryl Blue BGLN Liq 100 to Rainbow Trout LC₅₀  Values

Period of Exposure in hours	LC50  value with 95% cofidence limits (-) in mg/L
24	134   insufficient data
48	42      (35 - 52)
72	27      (20 - 37)
96	24      (19 - 30)

Validity criteria fulfilled:

not applicable

Conclusions:

The 96-h LC50 of the test item to Oncorhynchus mykiss under the conditions of the OECD test guideline 203 is 24.0 mg/L.

Executive summary:

In a 96-h acute toxicity study, Rainbow trout (Oncorhynchus mykiss) were exposed to the test item (40 % active ingredient) at nominal concentrations of 0, 10, 18, 32, 56, 100, 180 mg/L under static conditions. The 96-h NOEC was 10 mg/L. The LOEC was 18 mg/L with 20 % mortality after 96 h. The graphically derived 96-h LC₅₀was 24.0 mg/L. Sublethal effects were not documented. 

Based on the results of this study, the LC₅₀ for 100 % test item is calculated as 9.6 mg/L, leading to a classification as as Chronic 2 to O. mykiss in accordance with the CLP classification system.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for OECD TG 203 Fish acute toxicity study.

 

Results Synopsis

 

Test organism size: Length 51.5 – 52.0 mm and weight 1.33 – 1.47 g

Test Type: Semi-static

 

96 h LC₅₀:  24.0 mg/L (40 % act. ingr.)

NOEC: 10 mg/L (40 % act. ingr.)             

 

Endpoint(s) Effected: mortality

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

For details and justification of read-across please refer to the attached report in section 13 of IUCLID.

Reason / purpose for cross-reference:

read-across source

GLP compliance:

no

Analytical monitoring:

not specified

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

17.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: geometric mean value of LC0 and LC100

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: remark: calculated from geometric mean LC50 of the test material

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

30 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Table 1: Results of surviving fish and corresponding medium parameter measured.

Concentration (in mg/L)	Number of living fish after X- hours
	1	4	24	48	72	96	pH	O2
10.0	10	10	10	10	10	10	7.6	8
30.0	10	10	10	8	6	0	7.8	7.8
100.0	10	10	0				8	8.1
300.0	10	0					7.6	8.7

Validity criteria fulfilled:

not specified

Conclusions:

The 96-h EC50 of the test item to Danio rerio is 22 mg/L.

Executive summary:

In a 96-h acute toxicity study, zebrafish (Danio rerio) were exposed tothe test item (14.5 % active ingredient) at nominal concentrations of 10, 30, 100, and 300 mg/L. The 96-h NOEC was 10 mg/L. The 96-h LC₅₀ was 17.3 mg/L (geometric mean value of LC₀and LC₁₀₀).

Sublethal effects were not documented.  Based on the results of this study, the LC₅₀for 100 % test item would be calculated as 2.5 mg/L, leading to a classification as

classified as Chronic 2 to D. rerio in accordance with the CLP classification system.

 

Results Synopsis

Test organism size: not reported

Test Type: not reported

 

96 h LC₅₀:  22.0 mg/L (14.5 % act. ingr.)

96 h LC₀: 10 mg/L (14.5 % act. ingr.)           

 

Endpoint(s) Effected: mortality

This information is used in a read-across approach in the assessment of the target substance.

For details and justification of read-across please refer to the attached report in section 13 of IUCLID.

Reference 7

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

For details and justification of read-across please refer to the attached report in section 13 of IUCLID.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

8.78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

not specified

Conclusions:

The 96-h LC50 of the test item to Danio rerio is 8.78 mg/L.

Executive summary:

In a 96-h acute toxicity study, zebrafish (Danio rerio) were exposed to the test item at nominal concentrations of 3.0, 10, 30 and 100 mg/L. The 96-h NOEC was 3.0 mg/L. The 96-h LC₅₀was 8.78 mg/L. Sublethal effects were not documented.

Based on the results of this study the test item would be classified as Chronic 2 toD. rerio in accordance with the CLP classification system.

 

Results Synopsis

Test organism size: not reported

Test Type: not reported

 

96 h LC₅₀:  8.78 mg/L

 

Endpoint(s) Effected: mortality

This information is used in a read-across approach in the assessment of the substance to be registered.

For details and justification of read-across please refer to the attached report in section 13 of IUCLID.

Reference 8

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

For details and justification of read-across please refer to the attached report in section 13 of IUCLID.

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 42 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 24 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

9.6 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: not documented
- Mortality of control: 0
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

48 h LC50 = 42 mg/L (95 % CI: 35-52)
72 h LC50 = 27 mg/L (95 % CI: 20-37)
96 h LC50 = 24 mg/L (95 % CI: 19-30)
LC50 are estimated graphically and 95% Confidence limits calculated where possible, according to the method of GRanmo and Larsstuvold: Evaluation of results from short-term tests. The LC50 value and its confidence limits. Food and Agriculture Organisation of the United Nations TF-INT 1976. 237 p.

Sublethal observations / clinical signs:

During the test the time of survival was recorded and is presented in table 1. An overview of the LC₅₀ values for different observation times and the corresponding confidence limits is presented in table 2.

Table 1.       Toxicity of Yoracryl Blue RGLN Liq 100 to Rainbow Trout Median Periods of Survival

Concentration  (mg/L)	Median Period of Survival in minutes with 95% confidence limits  ( - )
180	922      ( 784 - 1084)
100	2167      (1954 - 2403)
56	1451      (976 - 2157)
32	4740      (3976 - 5651)
18	20 % mortality in 96 h
10	No mortality
0	No mortality

 

Table 2.          Toxicity of Yoracryl Blue BGLN Liq 100 to Rainbow Trout LC₅₀  Values

Period of Exposure in hours	LC50  value with 95% cofidence limits (-) in mg/L
24	134   insufficient data
48	42      (35 - 52)
72	27      (20 - 37)
96	24      (19 - 30)

Validity criteria fulfilled:

not applicable

Conclusions:

The 96-h LC50 of the test item to Oncorhynchus mykiss under the conditions of the OECD test guideline 203 is 24.0 mg/L.

Executive summary:

In a 96-h acute toxicity study, Rainbow trout (Oncorhynchus mykiss) were exposed to the test item (40 % active ingredient) at nominal concentrations of 0, 10, 18, 32, 56, 100, 180 mg/L under static conditions. The 96-h NOEC was 10 mg/L. The LOEC was 18 mg/L with 20 % mortality after 96 h. The graphically derived 96-h LC₅₀ was 24.0 mg/L. Sublethal effects were not documented. 

Based on the results of this study, the LC₅₀ for 100 % test item is calculated as 9.6 mg/L, leading to a classification as as Chronic 2 toO. mykiss in accordance with the CLP classification system.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for OECD TG 203 Fish acute toxicity study.

 

Results Synopsis

 

Test organism size: Length 51.5 – 52.0 mm and weight 1.33 – 1.47 g

Test Type: Semi-static

 

96 h LC₅₀:  24.0 mg/L (40 % act. ingr.)

NOEC: 10 mg/L (40 % act. ingr.)             

 

Endpoint(s) Effected: mortality

Description of key information

In a 96-h acute toxicity study by Munk 1987, individuals of L. idus were exposed to Basic Blue 41 methyl sulfate and a 96-h LC₅₀ was determined to be 68.1 mg/L. An additional study is available (Reust, 1980) reporting results for O. mykiss and D. rerio which were tested with a substance mixture containing 28.5 % of Basic Blue 41 methyl sulfate. 96-h LC₅₀ values, extrapolated to 100 % Basic Blue methyl sulfate, are 14.2 mg/L and 4.9 mg/L for O. mykiss and D. rerio, respectively.

Two additional studies are available (Reust, 1983; Reust, 1989) which investigated the effects of the structural analogue to D. rerio. LC₅₀values were 2.5 mg/L (extrapolated to 100 % test item)

and 8.78 mg/L, respectively. These values are again in the same range as the 96-h LC₅₀ value for D. rerio reported for Basic Blue methyl sulfate (Reust, 1980).

Although the detail level of the studies by Reust 1980, 1983 and 1989 are insufficient for assessment and are thus all rated as not reliable (Klimisch 4), the information is considered in a weight-of-evidence approach, demonstrating that D. rerio is more sensitive to Basic Blue 41 methyl sulfate compared to L. idus and O. mykiss.

Thus the lowest available 96-h LC₅₀value for D. rerio of 2.5 mg/L (Reust, 1983) is carried forward in the risk assessment.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

2.5 mg/L

Additional information

Information on Basic Blue 41 chloride is used in a read-across approach in the assessment of the substance to be registered.

For details and justification of read-across please refer to the attached report in section 13 of IUCLID.