N-methoxy-1-(2,4,6-trichlorophenyl)propan-2-amine

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Remarks:

The acute toxicity fish study was conducted solely to comply with a non-EU national registration requirement, and has been provided here in accordance with REACH, Article 22(1)e.

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 April 2015 to 14 October 2015

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The study was conducted under a National quality assurance scheme (HJ/T 155-2004) similar to OECD GLP.

Qualifier:

according to guideline

Guideline:

other: HJ!f 153-2004, The guidelines for the testing of chemicals [S]. Beijing: SEPA, 2004

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: CRC-MEP. The Guidelines for the Testing of Chemicals, Effects on Biotoc Systems [M]. 2nd edition. Beijing: China Environment Press. 2013: 30-36.

Version / remarks:

2013

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: GB/T 27861-2011, Chemicals-Fish acute toxicity test, Beijing: SAC, 2011.

Version / remarks:

2011

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: GB/T 29763-2013, Chemicals-Rare Minnow (Gobiocypris rarus) acute toxicity test Beijing: SAC, 2013.

Version / remarks:

2013.

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

no

Remarks:

The study was conducted under a National quality assurance scheme (HJ/T 155-2004) similar to OECD GLP.

Analytical monitoring:

yes

Details on sampling:

Sampling and analysis of the test solution
Water samples were taken (at least in duplicate) from each concentration during the definitive test at 0, 24, 48, 72 and 96 h. On each occasion, one sample was analysed after certain pre-treatments; the remaining samples were retained in case further analysis would be required.
Pre-treatment method of the sample
50.0 ml collected water samples were diluted to 100 mL with mobile phase (2-folds diluted), and filtrated by 0.22 fill1 millipore membrane (discarding the initial 10.0 mL filtrates), then the flltrates were analysed by UPLC-PDA.

Vehicle:

no

Details on test solutions:

In the range-finding test, the test solution was prepared by directly adding appropriate amounts of CA5204A in dilution water and then facilitating its dispersion by ultrasonication and stirring for 30 minutes. The amount of the test substance added was converted according to the purity of the test substance. The insoluble test substance settled at the bottom of the solution.
In the definitive test, the test medium was prepared as a slow stir super stock solution. The super stock solution (1 00 mg/L) was prepared by adding 0.3231 g CA5204A in 3 L dilution water. The aqueous test substance mixture was stirred for 24 hrs on a magnetic stirplate and a telfon stirbar at 35°C. The vortex height was set at least 10% of the static liquid height. At the end of the 24 h, stirring was stopped and the test substance was settled down to the bottom. The stock solution stands for 1 hour at room temperature prior to the removal of any undissolved test item by filtration through 0.45 micrometer millipore membrane (first approximate 100 mL discarded) to produce a 100% v/v stock solution of the test item.

Test organisms (species):

other: Gobiocypris rarus

Details on test organisms:

The test species of Rare minnow, Gobiocypris rarus (Batch No.: F20150315G), were obtained from a fish supplier of Institute of Hydrobiology, Chinese Academy of Sciences.
Fish were held at least for 12 days in holding tanks supplied with a continuous flow of aerated water before being used for testing. Fish to be used in the test were held for 7 days in water of the quality and temperature to be used in the test.
A photoperiod of 16 hours light, provided by overhead fluorescent tubes, and 8 hours dark was maintained. The oxygen concentration was more than 80% of the air saturation value.
The fish were fed daily during the holding period on proprietary fish food. They were held without food for approximately 24 hours before being placed into the test vessels. The ingredients of the fish food are given as follows:
Crude Protein > 36.0%
Crude Fat > 2.0%
Crude Fibre < 3.0%
Crude Ash < 12.0%
Moisture < 10.0%
Characteristics of the fish food are measured at least twice a year by Jiangsu Provincial Center for Disease Prevention and Control.
During the holding period the tanks was inspected daily and any debris or unhealthy or dead fish removed.
After the 48 hour settling-in period, no mortality was observed in the following 7 days. So the batch of fish (Batch No.: F20 150315G) was accepted.
The average wet-weight and length of the fish used to the test was 0.226 g and 2.54 cm, and the relative standard deviation (RSD) was 8.66% and 7.07%, respectively

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

10 ~ 250 mg (CaC03)/L

Test temperature:

23 ±2 °C

pH:

6.0 to 8.5

Dissolved oxygen:

75% to 96%

Nominal and measured concentrations:

Nominal concentrations of 5%, 10%, 15%, 20% and 30% stock solution were used in the defmitive test.
Measured concentrations were 0.92, 1.85, 2.98, 3.85 and 6.43 mg/L, respectively

Details on test conditions:

Synchronously blank control was used in the test. No replicate was assigned for each treatment group and control group, while the initial number of testing fish was 10 for each group.
The test fish were randomly chosen and put in different test solutions after the temperature has been adjusted to the required value. This was done in 30 min.
During the test, the following conditions were maintained:
-Light: 16 hours photoperiod daily (light intensity: 1000 lux to 1500 lux);
-Temperature: 23.2°C to 23.SOC;
-Oxygen concentration: 75%- 96% of the air saturation; No aeration.
-Feeding: none.
At 3, 6, 24, 48, 72 and 96 h, the mortalities of the fish were recorded, and observations on individual behaviour were performed. Meanwhile, measurements of pH, dissolved oxygen and temperature were carried out and recorded daily.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, CAS: 7778-50-9; purity: > 99.8%; Lot number: 10011460037; Nanjing Chemical Reagent Co., Ltd.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.72 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.85 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

3.85 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

During the test period, all fish in the control and treatment of 5% stock solution (measured concentration was 2.98 mg/L) were alive and appeared normal. Effects (fish lying on side or back) occurred at nominal test concentrations of 10% stock solution (measured concentration was 1.85 mg/L). All fish were dead after 96 h-exposure at nominal test concentration of20% stock solution (measured concentration was 3.85 mg/L).

Results with reference substance (positive control):

- Results with reference substance valid: Yes
- LC50 (6h) = 473 mg/L
- LC50 (12h) = 410 mg/L
- LC50 (18h) = 344 mg/L
- LC50 (24h) = 277 mg/L

Sublethal observations / clinical signs:

During the test period, all fish in the control and treatment of 0.92 mg/L were alive and appeared normal.

Validity criteria fulfilled:

yes

Conclusions:

The results showed that under valid static test conditions, the 96 h-LC50 of CA5204A to fish (Rare minnow, Gobiocypris rarus) is 2.72 mg/L, with 95% confidence limit of 2.43 mg/L ~ 3.05 mg/L (based on the measured concentration). The NOEC (96 h-LC0) is 1.85 mg/L. The minimum concentration causing 100% mortality (96 h-LC100) is 3.85 mg/L.

Executive summary:

Under static conditions, the acute toxicity of test substance (CA5204A) to Rare mmnow (Gobiocypris rarus) was conducted according to: "The guidelines for the testing of chemicals" (HJ/f 153-2004); and with reference to Procedure 203 of the "Guidelines for Testing of Chemicals" of the OECD: "Fish, Acute Toxicity Test" (1992).

A range-finding test and then a definitive test were performed respectively. Nominal concentrations of 1.0, 10.0 and 100 mg/L were used in the range-finding test. While the nominal concentrations of 5%, 10%, 15%, 20% and 30% stock solution were used in the definitive test. Water samples taken from the blank control and the treatments in the definitive test were analysed. Concentrations of the test substance were quantified using UPLC-PDA of the measured concentrations were 0.92, 1.85, 2.98, 3.85 and 6.43 mg/L, respectively. The analytical results showed that the concentration of the test substance is consistent in the test medium throughout the 96-hour test period (deviation within 20 %). Thus a static procedure was reasonable.

In the range-fmding test, 5 fishes per treatment and no replicates were used. 10 fishes per treatment were used in the definitive test. The test fishes were exposed for 96 hours to the test solution.

During the test period, the pH values of the control medium and test medium were between 7.92 and 8.13, the Dissolved Oxygen (DO) values varied from 75% ~ 96% of the air saturation at the test temperature, and the temperature of the test mediums were maintained in the range of 23.1 oc to 23.5°C. The total hardness was in the range of 142 mg (CaC03)/L to 153 mg (CaC03)/L in the beginning of the test, and all fishes in the control group were normal. With the same conditions, K2Cr2O7 was used as the positive control substance, and the resulting 24 h-LC50 was 277 mg/L. So the study met the acceptability criteria prescribed by the protocol (pH: 6.0 ~ 8.5; dissolved oxygen concentration: > 60% of the air saturation value; total hardness: 10 ~ 250 mg (CaC03)/L; temperature: (23±2) °C; 24 h-LC50 of K2Cr2O7 in the range of 200 mg/L to 400 mg/L). Therefore the test was considered valid.

Description of key information

The results showed that under valid static test conditions, the 96 h-LC50 of the test substance to fish (Rare minnow, Gobiocypris rarus) is 2.72 mg/L, with 95% confidence limit of 2.43 mg/L ~ 3.05 mg/L (based on the measured concentration). The maximum tested concentration causing no mortality (96 h-LC0 or No Observed Effect Concentration) is 1.85 mg/L. The minimum concentration causing 100% mortality (96 h-LC100) is 3.85 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

2.72 mg/L