2,6-dichloro-N-phenylaniline

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

other information

Study period:

February 06 - March 17, 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented and reported study, conducted equivalent to internationally accepted technical guideline in recognized research organization.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

, 1992, static system

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,6-dichloro-N-phenylaniline

EC Number:

239-349-0

EC Name:

2,6-dichloro-N-phenylaniline

Cas Number:

15307-93-4

Molecular formula:

C12H9Cl2N

IUPAC Name:

2,6-dichloro-N-phenylaniline

Details on test material:

Identification Code: PBS 3247
Generic/Trade name: Dichlordiphenylamin
BatchNo.: EN 322165.36
Appearance: beige solid
Purity: not available
Solubility (in water): 0.0063 g/l
Received: 17/10/94
Storage: room temperature
Stability: 12/95

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples of each test concentration were drawn from the approximate center of the test vessels. They were taken immediately before the exposure and after 96 hours exposure and kept at -18°C to -22°C until analysis.

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- (Test) Water: dechlorinated tap water (carbon filter)
- Method: Stock solution: 0.30 g test substance and 0.74 g 1% TWEEN 80 (FLUKA 93781) in water were mixed with and made up to 1000 ml with water (additional ultrasonication for 20 min). Calculated amounts of the stock solution to produce the desired test concentrations were given into the water. In addition calculated amounts of vehicle were given to the test solutions to achieve identical vehicle concentrations except for the blank. The test solutions were equilibrated for 4.5 hours.
- Controls: blank (dechlorinated tap water) and vehicle control
- Chemical name of vehicle: polyoxy-ethylene-sorbitan-monooleate (TWEEN 80)
- Concentration of vehicle in test medium: stock solution: 7.4 mg/l, final test solutions including vehicle control: 0.2 mg/l
- Evidence of undissolved material: a slight deposit was observed in concentration 1.9 mg/l from the start of the test to 24h exposure. The test substance appeared homogeneously distributed in the test vessels during 48-96h exposure.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: P. Hohler / CH-4314 Zeiningen
- Length at study initiation (length definition, mean, range and SD): 51 mm (42 - 59 mm) based on 7 fish
- Weight at study initiation (mean and range, SD): 1.13 g (0.64 - 1.72 g) based on 7 fish
- Feeding during test: none

ACCLIMATION
- Acclimation period: 70 days
- Acclimation conditions: same as test
- Treatment: none

ADAPTATION
24 hours / no food 24 hours prior to exposure

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

177 mg CaCO3/l

Test temperature:

test start: 13.1 °C
24-96h: 12.6 - 13.1 °C

pH:

test start: 7.7 - 8.1
24-96h: 8.0 - 8.3

Dissolved oxygen:

test start: 98% - 106%
24-96h: 84% - 103%

Salinity:

not applicable (freshwater)

Nominal and measured concentrations:

nominal concentration: 0.86, 1.5, 2.8, 5.0, and 9.0 mg/l
measured at test start: 0.10, 0.17, 0.36, 0.61, and 1.1 mg/l
measured at test end: 0.13, 0.29, 0.59, 0.81, and 2.7 mg/l
arithmetric mean: 0.12, 0.23, 0.47, 0.71, and 1.9 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass aquaria of 20 liters (35/22/26 cm) filled with 15 liters
- Aeration: gentle aeration during exposure
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.53 g/l


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water (carbon filter)
- Culture medium different from test medium: no
- Intervals of water quality measurement: oxygen content, pH and temperature were measured at 0, 24, 48, 72 and 96 hours


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hour daily


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality after 2-4, 24, 48, 72 and 96 hours exposure; sublethal symptoms daily


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: results not reported

Reference substance (positive control):

not required

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.2 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

1.2 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

1.2 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 5

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.71 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 6

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 7

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.23 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

other: sublethal effects

Details on results:

- Effect data: Mortality occured in the highest concentration of 1.9 mg/l (nominal 9 mg/l) only (100% mortality).
- Behavioural abnormalities: fish showed sublethal effects (change in pigmentation, loss of equi1ibrium, change in the swimming behaviour and in the respiratory function) at concentrations >0.23 mg/l (nominal >1.5 mg/l).
- Mortality of control: 0
- Other adverse effects control: none

Reported statistics and error estimates:

The LC-5O values were calculated according to the maximum likelihood method, probit model (Finney, D.J., 1971), using the probit procedure in SAS Version 6.08 (SAS / STAT Users Guide, Vol. 2, SAS Institute Inc.).
LC50 (48, 72 and 96 h): slope: 16.5, goodness of fit X²: 0.0 DF=3, P=1.0
LC50 (24 h): slope: 7.37, goodness of fit X²: 0.0 DF=3, P=1.0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Validity Criteria: Mortality in controls <10%; dissolved oxygen conc. >60% air saturation; conc. of test substance sufficiently constant; conc. based on measured concentration because deviation from nominal concentration >20%.

Conclusions:

96h- LC50: >1 ≤10 mg/l

Executive summary:

In a 96-h acute toxicity study, rainbow trout (Oncorhynchus mykiss) were exposed to PBS 3247 under static conditions in accordance with OECD Guideline 203 (Fish Acute Toxicity Test) and in compliance with GLP. Reliability grade 1 was assigned. Nominal concentrations of 0 (control and solvent control), 0.86, 1.5, 2.8, 5.0, and 9.0 mg/l were tested. The results were based on arithmetic mean concentrations of 0.12, 0.23, 0.47, 0.71, and 1.9 mg/l.

LC 50 (96 h) calculated:	1.2 mg/l
95 % confidence limit:	none
Slope:	16.5
Goodness of fit X²:	0.0  DF= 3 P=1.0
NOEC (96 h):	0.23 mg/l
LC 0 (96 h):	0.71 mg/l
LC 100 (96 h):	1.9 mg/l

Mortality occurred in the highest concentrations of 1.9 mg/l only (100% mortality). Fish showed sublethal effects at concentrations > 0.23 mg/l (change in pigmentation, loss of equilibrium, change in the swimming behaviour and in the respiratory function). No fish died in the control group.