o-phenylenediamine

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: 40 CFR 797.1300 Daphnid acute toxicity test

GLP compliance:

yes

Remarks:

40 CFR 160 and 40 CFR 792

Test material

Test material information

Constituent 1

Reference substance name:

o-phenylenediamine

EC Number:

202-430-6

EC Name:

o-phenylenediamine

Cas Number:

95-54-5

Molecular formula:

C6H8N2

IUPAC Name:

benzene-1,2-diamine

Details on test material:

- Purity: 99.8%

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: collected in 1.5 mL autosampler vials
- Sample storage conditions before analysis: Immediately frozen in liquid nitrogen and stored at -70°C

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: Haskell Laboratory
- Age at study initiation (mean and range, SD): Less than 24 hours old
- Method of breeding: Parent Daphnia magna were reared at Haskell Laboratory in 250 mL Pyrex beakers (1 per beaker) which contained 200 mL of (autoclaved) FFTWater held at approximately 20°C. Daphnids were fed 2 algal species, Ankistrodesmus falcatus and Selenastrum capricornutum, at a rate of 250000 cells/mL of each species, three times weekly during water renewal. Neonates were collected from 12 day old parent daphnids for use in this test.
- Feeding during test: Yes
- Food type: Ankistrodesmus falcatus and Selenastrum capricornutum
- Amount: 250000 cells/mL of each species
- Frequency: At test start and daily at test solution renewal

ACCLIMATION
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: See above
- Feeding frequency: See above
- Health during acclimation (any mortality observed): No sickness, injury or abnormality was observed

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

82 mg/L as CaCO3

Test temperature:

Mean: 20.5°C; range: 20.2 to 20.7°C

pH:

7.6 - 8.0

Dissolved oxygen:

9.0 - 9.1

Nominal and measured concentrations:

Nominal: 0.041, 0.10, 0.26, 0.64, 1.6, 4.0, 10 mg/L
Mean measured: 0.022, 0.072, 0.22, 0.57, 1.8, 3.6, 9.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: Pyrex beakers, 250 mL, 7 cm diameter, 200 mL test solution, 6 cm depth
- Aeration: No

- Renewal rate of test solution (frequency/flow rate): At 24 hours
- 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: Haskell Laboratory well water flowed through aquaria containing fathead minnows and subsequently filtered (= filtered fish tank water, FFTW)
- Total organic carbon: <0.5 and 1 mg/L (sampled twice during experiment)
- Particulate matter: <4 mg/L
- Metals: (all data expressed as mg/L) Al: <0.2 - Sb: <0.2 - As: <0.002 - Be: <0.01 - B: <0.04 - Ca: 24.8 - Cd: <0.01 - Cr: <0.05 - Co: <0.05 - Cu: <0.02 - Fe: 0.1 - Pb: <0.1 - Mg: 3.9 - Hg: <0.00020 - Ni: <0.05 - K: 1.9 - Se: <0.002 - Ag: <0.02 - Na: 8.4 - Zn: 0.1 (Zn concentration probably due to contamination during sampling.)
- Pesticides: ND
- Chlorine: 6 mg/L chloride
- Alkalinity: 86 mg/L as CaCO3
- Conductivity: 180 µmhos/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: D.O., pH and temperature were measured at start, 24 hours, renewal and 48 hours


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light, 8 hours darkness. Approximately 25 minutes of transitional light
- Light intensity: 118-226 Lux

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
- Results used to determine the conditions for the definitive study: This study served as a range finding study for further experiments

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.4 mg/L

Nominal / measured:

meas. (geom. mean)

Basis for effect:

mobility

Remarks on result:

other: 95% Confidence Limits: 0.91 - 2.1 mg/L

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (geom. mean)

Basis for effect:

mobility

Remarks on result:

other: 95% Confidence Limits: 1.5 - 2.3

Reported statistics and error estimates:

Moving average-angle technique (Peltier WH, Weber CI. 1985. Methods for measuring the acute toxicity of effluents to freshwater and marine organisms. EPA/600/4-85/013.)

Any other information on results incl. tables

Observed Immobility in Percent

Mean Measured Concentration (mg/L)	24 Hours		48 Hours
	Replicate A	Replicate B	Replicate A	Replicate B
Control	0	0	0	0
0.022	0	0	0	0
0.072	0	0	0	0
0.22	10	10	10	10
0.57	0	0	0	0
1.8	40	40	80	70
3.6	100	100	100	100
9.0	100	100	100	100

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

This study established a 48-hr EC50 in daphnids of 1.4 mg/L.

Executive summary:

oPDA was tested with Daphnia magna in an unaerated, static, 48-hour test with test solution renewal at 24 hours. Nominal concentrations were 0.041, 0.10, 0.26, 0.64, 4.0 and 10 mg/L. Mean measured concentrations were 0.022, 0.072, 0.22, 0.57, 1.8, 3.6, and 9.0 mg/L, respectively. The 48-hour EC50 based on mean measured concentrations and immobility was 1.4 mg/L.