3-aminophenol

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From August 26, 2013 to January 29, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed according to the standard guideline.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Principles of method if other than guideline:

Study was conducted to evaluate the toxic nature of test chemical on the growth of aquatic invertebrates in accordance with OECD guideline 202.

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1.0, 0.45, 0.21, 0.093 and 0.042 mg test item/L
- Sampling method:One sample of the stock solution and duplicate samples from the freshly prepared test media of all test concentrations and the control
were taken at the start of the test and at day 1.
- Sample storage conditions before analysis: All samples were diluted by a factor of two with acetonitrile, directly after sampling. All samples were stored in a freezer (≤ - 10 °C), protected from light until analysis was performed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Before test start and before the test medium renewal a concentrated stock solution was prepared by dissolving 20.05 and 20.24 mg test item into 501.25 and 506 mL test water by intense stirring for 10 minutes. Adequate volumes of this test medium were diluted with test water to prepare the test media. The test media were prepared just before introduction of the daphnids (= start of the test) and test medium renewal on Day 1.
- Eluate: no
- Differential loading: no
- Controls: In the control, test water was used without addition of the test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not used
- Concentration of vehicle in test medium (stock solution and final test solution):
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no. The appearance of the test item in the test media was observed at the start of the test, on Day 1 in the old and the new test media and at the end of the test in the test media of all test concentrations.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Straus)
- Strain: clone 5
- Source: IBACON's in-house laboratory culture.
- Age at study initiation (mean and range, SD): From 2 to 18 hours old
- Method of breeding: The daphnids were bred in the laboratories of IBACON under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in the same reconstituted water as the test water used in the test. The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of IBACON.
- Feeding during test: No

ACCLIMATION
- Acclimation period: Was not necessary, since the test was performed in the same medium as the culturing.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

2.5 mmol/L (= 250 mg/L) as CaCO3

Test temperature:

20 to 21 °C in the freshly prepared media;
20 °C in the aged test media

pH:

7.8 to 7.9 in the freshly prepared media;
7.7 to 7.8 in the aged test media; and thus the pH-value did not vary by more than 1.5 units

Dissolved oxygen:

8.5 to 9.3 mg/L in the freshly prepared media;
8.1 to 8.3 mg/L in the aged test media

Nominal and measured concentrations:

1.0, 0.45, 0.21, 0.093 and 0.042 mg test item/L, corresponding to geometric mean measured concentrations of 0.99, 0.387 and 0.164 mg test item/L (the two lowest concentrations were below LOQ).

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers
- Type (delete if not applicable): closed; covered with a lid to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions.
- Material, size, headspace, fill volume: Glass; 100 mL volume containing approximately 60 mL of test medium.
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): renewal on Day 1
- No. of organisms per vessel: 20 daphnids per control and test concentration, divided into 4 groups of 5 animals, each group in 60 mL test medium
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): na

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted Water (Elendt "M4"). Analytical grade salts and additives were added in deionised water (conductivity < 5 μScm-1)

- Total organic carbon: nd
- Alkalinity: 0.9 mmol/L
- Ca/mg ratio: 2.5 mmol/L (= 250 mg/L) as CaCO3
- Conductivity: < 5 μScm-1
- Culture medium different from test medium: No
- Intervals of water quality measurement: The water temperature, pH-values and the dissolved oxygen concentrations were determined in all freshly prepared and aged test media of each treatment group.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light : 8 h dark
- Light intensity: 810 - 1080 lux (measured once during the test)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility: The mobility of the daphnids was determined by visual observation after 24 and 48 hours. Those animals that are not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile (even if they could still move their antennae).

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Justification for using less concentrations than requested by guideline: na
- Range finding study: yes
- Test concentrations: not in GLP, not reported in the present study
- Results used to determine the conditions for the definitive study: not reported in the present study

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.267 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.164 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no
- Observations on body length and weight: nd
- Other biological observations: no
- Mortality of control: 0%
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no; clear test medium.
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: In the most recent test with the reference item potassium dichromate performed in July 2013, the EC50 after 24 hours was determined to be 1.479 mg test item/L indicating that the sensitivity of the Daphnia was consistent with the level proposed by the OECD 202 guideline (EC50-24 h between 0.6 and 2.1 mg potassium dichromate/L).

Reported statistics and error estimates:

The 24-hour and 48-hour EC50, EC20 and EC10 and the 95 % confidence limits were calculated by Probit analysis.
The NOEC and LOEC after 24 and 48 hours were determined directly from the raw data.
The software used to perform the statistical analysis was ToxRat Professional, Version 2.10.05, ToxRat Solutions GmbH.

Influence of the test item on the Mobility of Daphnia magna:

Nominal Concentration	No. of Daphnia tested	No. of immobilised Daphnia after		% of immobilised Daphnia after
[mg test item/L]	20	24 h	48 h	24 h	48 h
control	20	0	0	0	0
0.042	20	0	0	0	0
0.093	20	0	0	0	0
0.21	20	0	0	0	0
0.45	20	2	16	10	80
1.0	20	5	20	25	100

Summary of Analytical Results

sample description [mg test item/L]	% of nominal^1	RSD [%]	n
Control	n.a. (<LOQ)	n.a.	8
0.042	n.a. (<LOQ)	n.a.	8
0.093	n.a. (<LOQ)	n.a.	8
0.21	78	5	8
0.45	86	12	8
1	99	6	8

1 mean value of all measured samples per treatment group

RSD: relative standard deviation per treatment group

n: number of analysed samples

n.a.: not applicable

Validity criteria fulfilled:

yes

Remarks:

see above

Conclusions:

The toxic effect of the test item to Daphnia magna was assessed in a semi-static dose-response test. The 48-hour NOEC was determined to be 0.164 mg test item/L. The 48-hour LOEC was determined to be 0.387 mg test item/L and the 48-hour EC50 value was calculated to be 0.267 mg test item/L.

Executive summary:

The Acute Toxicity of the test item to Daphnia magna was determined in a Semi-Static 48-hour Immobilisation Test, according to OECD 202 guideline and following GLP. This study encompassed 6 treatment groups (5 dose rates of the test item, control) each containing 20 individuals. The mobility of the daphnids was determined in a semi-static 48-hour test by visual observation after 24 and 48 hours. 1.0, 0.45, 0.21, 0.093 and 0.042 mg test item/L, corresponding to geometric mean measured concentrations of 0.99, 0.387 and 0.164 mg test item/L (the two lowest concentrations were below LOQ) and a control were tested. Water temperature: 20 to 21 °C; pH value: 7.7 to 7.9; dissolved oxygen concentration: 8.1 to 9.3 mg/L; photoperiod: 16 h light - 8 h dark; light intensity: 810 - 1080 lux; and thus were within the ranges requested by guideline OECD 202. Additionnally all the validity criteria are fulfilled according the oecd 202 guideline. After 48 hours of exposure no immobilisation of the test animals was observed in the control and up to and including the test item concentration of 0.164 mg test item/L. At the concentration of 0.387 mg test item/L, 16 animals were immobile and 20 animals were immobile at the concentration of 0.99 mg test item/L. The quantification of the test item was performed using liquid chromatography (HPLC-UV). In the both lowest test concentrations of nominal 0.042 and 0.093 mg test item/L all measured values are below the limit of quantification of the analytical method. In the freshly prepared test media 90% of the nominal test concentration was found (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L). In the 24 hours old test media 85% of the nominal value (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L) was determined. During the test the daphnids were exposed to a mean of 88% of nominal (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L). All reported biological endpoints refer to mean measured concentrations. For the concentration levels of 0.042 and 0.093 mg test item/L, which were below the limit of quantification (LOQ), 50% of the LOQ (= 0.01 mg test item/L) were introduced for the statisitical evaluation of the biological endpoints (as recommended in OECD No.23). Based on the effect concentration it was consider that the chemical was toxic to the mobility of daphnia magna and consider to be classified as aquatic acute 1 as per the CLP classification criteria.

Description of key information

The Acute Toxicity of the test item to Daphnia magna was determined in a Semi-Static 48-hour Immobilisation Test, according to OECD 202 guideline and following GLP. This study encompassed 6 treatment groups (5 dose rates of the test item, control) each containing 20 individuals. The mobility of the daphnids was determined in a semi-static 48-hour test by visual observation after 24 and 48 hours. 1.0, 0.45, 0.21, 0.093 and 0.042 mg test item/L, corresponding to geometric mean measured concentrations of 0.99, 0.387 and 0.164 mg test item/L (the two lowest concentrations were below LOQ) and a control were tested. Water temperature: 20 to 21 °C; pH value: 7.7 to 7.9; dissolved oxygen concentration: 8.1 to 9.3 mg/L; photoperiod: 16 h light - 8 h dark; light intensity: 810 - 1080 lux; and thus were within the ranges requested by guideline OECD 202. Additionnally all the validity criteria are fulfilled according the oecd 202 guideline. After 48 hours of exposure no immobilisation of the test animals was observed in the control and up to and including the test item concentration of 0.164 mg test item/L. At the concentration of 0.387 mg test item/L, 16 animals were immobile and 20 animals were immobile at the concentration of 0.99 mg test item/L. The quantification of the test item was performed using liquid chromatography (HPLC-UV). In the both lowest test concentrations of nominal 0.042 and 0.093 mg test item/L all measured values are below the limit of quantification of the analytical method. In the freshly prepared test media 90% of the nominal test concentration was found (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L). In the 24 hours old test media 85% of the nominal value (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L) was determined. During the test the daphnids were exposed to a mean of 88% of nominal (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L). All reported biological endpoints refer to mean measured concentrations. For the concentration levels of 0.042 and 0.093 mg test item/L, which were below the limit of quantification (LOQ), 50% of the LOQ (= 0.01 mg test item/L) were introduced for the statisitical evaluation of the biological endpoints (as recommended in OECD No.23). Based on the effect concentration it was consider that the chemical was toxic to the mobility of daphnia magna and consider to be classified as aquatic acute 1 as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.267 mg/L

Additional information

Based on the experimental data from various database for the target chemical study have been reviewed to determine the mode of action of 3-aminophenol on the mobility rate and behavior of aquatic invertebrates. The studies are as mentioned below:

 

In the first key study from experimental report 2015. The Acute Toxicity of the test item to Daphnia magna was determined in a Semi-Static 48-hour Immobilisation Test, according to OECD 202 guideline and following GLP. This study encompassed 6 treatment groups (5 dose rates of the test item, control) each containing 20 individuals. The mobility of the daphnids was determined in a semi-static 48-hour test by visual observation after 24 and 48 hours. 1.0, 0.45, 0.21, 0.093 and 0.042 mg test item/L, corresponding to geometric mean measured concentrations of 0.99, 0.387 and 0.164 mg test item/L (the two lowest concentrations were below LOQ) and a control were tested. Water temperature: 20 to 21 °C; pH value: 7.7 to 7.9; dissolved oxygen concentration: 8.1 to 9.3 mg/L; photoperiod: 16 h light - 8 h dark; light intensity: 810 - 1080 lux; and thus were within the ranges requested by guideline OECD 202. Additionnally all the validity criteria are fulfilled according the oecd 202 guideline. After 48 hours of exposure no immobilisation of the test animals was observed in the control and up to and including the test item concentration of 0.164 mg test item/L. At the concentration of 0.387 mg test item/L, 16 animals were immobile and 20 animals were immobile at the concentration of 0.99 mg test item/L. The quantification of the test item was performed using liquid chromatography (HPLC-UV). In the both lowest test concentrations of nominal 0.042 and 0.093 mg test item/L all measured values are below the limit of quantification of the analytical method. In the freshly prepared test media 90% of the nominal test concentration was found (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L). In the 24 hours old test media 85% of the nominal value (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L) was determined. During the test the daphnids were exposed to a mean of 88% of nominal (average of the nominal test concentrations of 0.21, 0.45 and 1.0 mg test item/L). All reported biological endpoints refer to mean measured concentrations. For the concentration levels of 0.042 and 0.093 mg test item/L, which were below the limit of quantification (LOQ), 50% of the LOQ (= 0.01 mg test item/L) were introduced for the statisitical evaluation of the biological endpoints (as recommended in OECD No.23). Based on the effect concentration it was consider that the chemical was toxic to the mobility of daphnia magna and consider to be classified as aquatic acute 1 as per the CLP classification criteria.

 

First key study was supported by the second study from authoritative database 2018. Acute Immobilization effect of test chemical to Daphnia magna was studied in accordance with the OECD Guideline 202 (Daphnia sp., Acute Immobilisation on Test and Reproduction Test). Test conducted in the semi-static system for the total exposure period of 48hrs. Sample analyzed by the HPLC with the frequency of analysis at the start of exposure, before renewal of test solution. Toxicity measured at the nominal concentration 10, 4.0, 1.60, 0.640, 0.256, 0.102 mg/L (geometric ratio 2.5), control. 100 mL per vessel test solution volume was added, on 5 daphnia magna per vessel (20 per concentration) toxicity was checked. Results after 24-hour exposure at 50 % Effective concentration (EC50) for immobilization was 1.79 mg/L (95 % CL: 1.28 - 2.57 mg/L). After 48-hour exposure No observed effect concentration (NOEC) for immobilization was 0.256 mg/L, and Lowest concentration causing 100 % immobility at 1.60 mg/L. Based on the immobility of daphnia magna due to the exposure of chemical m-Aminophenol, the EC50 for 48hrs of exposure was 0.447 mg/l. Based on the EC50, it can be concluded that the chemical was toxic and be consider to be classified in aquatic acute category 1.

 

 

Similarly damaging effects of environmental chemicals (from peer reviewed journal 1989) in the Daphnia test as a basis for evaluation of the environment hazard in aquatic systems was measured. The effect of potential pollutants on the ability of Daphnia magna Straus to swim was carried out in accordance with the DIN 38412, Part II. Test conducted under the static system. From the solutions with the pollutants, dilution series were prepared using "dilution water". The concentration steps of the test solution were selected so as to give 3--4 EC values in a range between EC 0 and EC 100. Effect measured on the 6-24 hours old daphnia magna Straus which were not feed during the time of testing. 50 ml beakers and two 50 ml beakers with ground-glass stoppers were used. Loading amounted to one test animal per 2 ml test medium as ten 6-24 h old daphnids were placed in each test and control vessel, i.e. 20 animals per concentration step. Based on the mobility of daphnia magna by the exposure of chemical 3-aminophenol, in the interval of 24hrs and 48 hrs effect were measured. After 24 and 48 hrs effect (EC50) was observed at 2.2 mg/l and 1.1 mg/l. based on the EC50, chemical was consider as toxic to aquatic environment and classified as aquatic chronic 2 as per the CLP classification criteria.

 

As the maximum experimental studies performed according to the OECD guidelines supports the toxic and hazardous nature of test chemical thus on basis chemical 3-aminophenol was consider as toxic to aquatic invertebrates and classified as aquatic acute 1 as per the CLP classification criteria.