2,2'-phenyliminodiethanol

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Test performed in February 1995.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

only basic data given; legally binding classification (KBwS, WGK 2)

Qualifier:

according to guideline

Guideline:

other: German Industrial Standard DIN 38412-11

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL- Source and lot/batch No.of test material: Merck (Name in KBWS database: N,N-Di(2hydroxyethyl)anilin)- Expiration date of the lot/batch: no data- Purity test date: no data

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM - Common name: Waterflea - Strain/clone: clone 5 - Source: Laboratory - Age of parental stock (mean and range, SD): not specified; culture: fed with Desmodesmus subspicatus in CHU medium - Age at study initiation: < 24 h - Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20±1 °C

pH:

dilution water: 7.86test medium: 7.83

Dissolved oxygen:

dilution water: 99%

Nominal and measured concentrations:

nominal: 0 (control), 12.60, 17.64, 24.96, 34.57, 48.40, 67.76, 94.86, 132.81, 185.93, 260.31, 364.43, 510.20, 714.29, and 1000 mg/L

Details on test conditions:

TEST SYSTEM - Test vessel: not specified - No. of organisms per vessel: 10 - No. of vessels per concentration (replicates): 2 - No. of vessels per control (replicates): 2 TEST MEDIUM / WATER PARAMETERS - Source/preparation of dilution water: reconstituted freshwater according to German Industrial Standard DIN 38412, part 11 OTHER TEST CONDITIONS - Adjustment of pH: no - Photoperiod: continuous darkness EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : - immobilisation: after 24 and 48 h- pH and dissolved oxygen: after 48 h

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

87.85 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

279.52 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 5% - Effect concentrations exceeding solubility of substance in test medium: no

Immobilisation, pH and dissolved oxygen

Test item conc. (mg/L)	Cumulative number of immobile daphnids after				pH	Dissolved oxygen
	24 h		48 h		48 h	48 h
	n	%	n	%	-	%
control	0	0%	1	5%	7.62	97%
12.60	11	55%	20	100%	7.88	95%
17.64	13	65%	20	100%	7.87	94%
24.69	10	50%	20	100%	7.91	96%
34.57	12	60%	20	100%	7.87	94%
48.40	11	55%	18	90%	7.87	95%
67.76	10	50%	15	75%	7.87	95%
94.86	6	30%	12	60%	7.88	96%
132.81	1	5%	6	30%	7.87	96%
185.93	0	0%	0	0%	7.86	96%
260.31	0	0%	0	0%	7.86	96%
364.43	0	0%	0	0%	7.85	97%
510.20	0	0%	0	0%	7.80	97%
714.29	0	0%	0	0%	7.75	96%
1000	14	70%	20	100%	7.92	95%

Validity criteria according to OECD TG 202

For a test to be valid, the following performance criteria apply:

- In the control, including the control containing the solubilising agent, not more that 10 percent of the daphnids should have been immobilised: Fulfilled (Mortality of control: 5%)

- The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/l in control and test vessels: Fulfilled (see the Table above).

Validity criteria fulfilled:

yes

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

July 1996 - October 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Updating Nov. 1989.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL- Source and lot/batch No.of test material: BASF AG, 59-7325 (date of production: February 1988)- Expiration date of the lot/batch: not specified in study report- Purity test date: characterization at ZAX under 96L00222OTHER SPECIFICS:- Analytical purity: 97.7%- Impurities (identity and concentrations): 1.5% N,N-Bis-(2-Hydroxyethyl)-aniline mono ethylether; 0.4% N-(2-Hydroxyethyl) aniline; 0.08% water

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control), 3.13, 12.5, and 100 mg/L - Sampling method: samples from vessels without daphnids taken at 0 and 48 h; samples from vessels with daphnids taken after 48 h - Sample storage conditions before analysis: refrigerator

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)- Method: For the preparation of the stock solution (100 mg/L, nominal) the test substance was melted at 80 °C and after that it was stirred for about 10 minutes at 22±1 °C in M4-medium.- Differential loading: yes- Controls: negative control- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): not applicable- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM- Common name: Waterflea- Strain: Institut National de Recherche Chimique Appliquée, France- Source: originally supplied from 'Institut National de Recherche Chimique appliquee, France, and has been bred from 1978 in the Laboratories of BASF - Age of parental stock (mean and range, SD): 2 to 4 weeks- Age at study initiation (mean and range, SD): 2 to 24 h- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

total hardness: 2.40 mmol/L

Test temperature:

19.9 to 21.1 °C

pH:

test start: 8.0 to 8.1test end: 7.9

Dissolved oxygen:

test start: 8.1 to 8.2 mg/Ltest end: 7.7 to 7.8 mg/L

Conductivity:

607 µS/cm (test medium)

Nominal and measured concentrations:

nominal: 0 (control), 3.13, 6.25, 12.5, 25, 50, and 100 mg/L
measured: recovery: 98.4% to 105.4%

Details on test conditions:

TEST SYSTEM- Test vessel: reagent tubes with flat bottom- Material, size, headspace, fill volume: glass, 20-mL total volume, 10 mL fill volume- No. of organisms per vessel: 5- No. of vessels per concentration (replicates): 4- No. of vessels per control (replicates): 4TEST MEDIUM / WATER PARAMETERS- Source/preparation of dilution water: according to guideline (synthetic freshwater prepared on the basis of clean and deionized water with a conductivity of < 0.05 µS/cm)- Alkalinity: up to pH 4.3: 0.84 mmol/L- Ca/Mg ratio: 4:1- Culture medium different from test medium: noOTHER TEST CONDITIONS- Adjustment of pH: no- Photoperiod: 16/8 h light/dark - Light intensity: About 5 - 6 µE/(m²s) in the range of 400 - 700 nm (Artificial Iight, OSRAM L58 W31 warm white)EFFECT PARAMETERS MEASURED (with observation intervals if applicable): - temperature: After 0, 24, and 48 h- pH values and dissolved oxygen: After 0 and 48 hours- Mobile daphnids: after 0, 3, 6, 24 and 48 hoursTEST CONCENTRATIONS- Spacing factor for test concentrations: 2

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

94.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

analytically verified

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 75.8 to 117.5 mg/L

Immobilisation

Test item conc. (mg/L, nominal)	Immobilised animals after
	3 h	6 h	24 h	48 h
0	0	0	0	1
3.13	0	0	0	0
6.25	0	0	0	0
12.5	0	0	0	3
25	0	0	0	3
50	0	0	1	5
100	0	0	4	11

 

pH and dissolved oxygen

Test item conc. (mg/L, nominal)	pH		Dissolved oxygen (mg/L)
	0 h	48 h	0 h	48 h
0	8.1	7.9	8.2	7.8
3.13	8.0	7.9	8.1	7.7
6.25	8.0	7.9	8.2	7.7
12.5	8.0	7.9	8.2	7.7
25	8.0	7.9	8.2	7.7
50	8.0	7.9	8.2	7.7
100	8.0	7.9	8.2	7.7

 

Analytical verification of test item concentrations

Test item conc.	Measured concentration
	0 h		48 h
Stocked with daphnids	no		no		yes
(mg/L, nominal)	(mg/L)	(% of nominal)	(mg/L)	(% of nominal)	(mg/L)	(% of nominal)
0	< 2	n.d.	< 2		< 2
3.13	3.1	99.0%	3.1	99.0%	3.3	105.4%
12.5	12.7	101.6%	12.3	98.4%	12.7	101.6%
100	99.8	99.8%	98.5	98.5%	99.6	99.6%

Validity criteria according to OECD TG 202

For a test to be valid, the following performance criteria apply:

- In the control, including the control containing the solubilising agent, not more that 10 percent of the daphnids should have been immobilised: Fulfilled (Mortality of control: 5%)

- The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/l in control and test vessels: Fulfilled (see the Table above).

Validity criteria fulfilled:

yes

Description of key information

Acutely harmful to aquatic invertebrates

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

87.85 mg/L

Additional information

The effect of 2,2 -phenyliminodiethanol (CAS 120 -07 -0) was tested according to German Industrial Standard DIN 38412 -11 for the German Commission for the assessment of substances hazardous to water (KBWS, 1995). The study was performed without GLP and analytical verification of test item concentrations. Test concentrations can be assumed to be stable over the exposure period based on the results of the second daphnid study by BASF AG (1998). The test protocol only contains basic informations on the test conditions and the performance. The 48 -h EC50 was determined to be 87.85 mg/L (95% CL 72.33 to 106.70 mg/L; nominal).

The assessment of the toxicity to microorganisms is based on effect data taken from the “Commission for the Evaluation of Substances Hazardous to Waters” (KBWS) data sheet no. 1385, which is part of the German Federal Water Act (Article 19g). Based on the collected data a substance is classified according to its water-hazardous properties (WGK 1 to 3). 2,2 -phenyliminodiethanol was classified as WGK 2, which indicates a hazard to waters. As the list with the classified substances was published in the Bundesanzeiger (Federal Ministry of Justice, 2005. Bundesanzeiger No. 142a), the classification is legally binding for the industry.

In addition, a 48-h EC50 of 98.4 mg/L (95% CL 75.8 to 117.5 mg/L; measured) was determined in a GLP study testing the effect of 2,2'-phenyliminodiethanol (CAS 120 -07 -0) on the mobility of Daphnia magna. The study was conducted according to EU method C.2 (BASF AG, 1998, report no. 96/0282/50/1). The test item concentrations were analytically verified. The measured concentrations were in good agreement with the nominal values, which indicates a good stability of the test item. The EC50 is within the same order of magnitude as the EC50 determined by KBWS.

As the KBWS study (1995) has a higher sensitivity than the study performed by BASF (1998) it was selected as key study, although it has alower reliability (RL 2, basic data given, no analytical verification of test item concentrations). The study by BASF (1998) was selected as supporting study, although it has a higher reliability (RL 1; GLP guideline study).