[ethane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

from 16 May 1979 to 30 May 1979

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study was conducted according to an appropriate national standard method. It was compliant with GLP and analytical monitoring was carried out

Qualifier:

according to guideline

Guideline:

other: EPA-660/3-75-009 (US EPA)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations at days 0, 2, 5, 7, 9 and 12

- Sampling method: the sample collection method is not reported, however samples were collected each Monday before water renewals to determine the stability of the substance.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: the test concentrations were prepared by transferring direct weights of the test substance to the test chambers.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM

- Common name: rainbow trout

- Source: Spring Creek hatchery in Lewiston, Montana, USA

- Length at study initiation (length definition, mean, range and SD): mean standard length 46.2 mm

- Weight at study initiation (mean and range, SD): mean weight 1.77 g

- Method of breeding: not reported

- Feeding during test

- Food type: commercial fish food (Rangen's)

- Amount: 3% body weight

- Frequency: daily


ACCLIMATION

- Acclimation period: 14 day

- Acclimation conditions (same as test or not): culture techniques as described by Brauhn et al.

- Type and amount of food: same as during test

- Feeding frequency: same as during test

- Health during acclimation (any mortality observed): <5% mortalities

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

14 d

Hardness:

255 ppm as CaCO3

Test temperature:

12⁰C

pH:

control range: 7.2 to 8
low dose medium range: 7.2 to 8.1
high dose range: 6.7 to 7.8

Dissolved oxygen:

>60%

Nominal and measured concentrations:

Nominal concentrations: 0, 37, 65, 120, 210 and 370 mg active acid/L.
Measured concentrations: 0, 35, 62, 114 , 205 and 345 mg active acid/L.

Details on test conditions:

TEST SYSTEM

- Test vessel: aquaria

- Material, size, headspace, fill volume: 40 L glass aquaria filled with 30 L of test solution

- Aeration: yes

- Renewal rate of test solution (frequency): 75% media was replaced on Monday, Wednesday and Friday

- No. of organisms per vessel: 30

- No. of vessels per concentration (replicates): 0

- No. of vessels per control (replicates): 0


TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: laboratory well water

- Alkalinity: 369 ppm as CaCO3

- Conductivity: 50 uhmos/cm

- Culture medium different from test medium: no

- Intervals of water quality measurement: water quality measurements were done on each renewal date in the control, low and high concentrations.


OTHER TEST CONDITIONS
- Adjustment of pH: none reported
- Photoperiod: 16 h daylight
- Light intensity: not reported


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : the fish were observed for mortality and abnormal behaviour initially and once every 24 hours for the 14 day period.


TEST CONCENTRATIONS

- Spacing factor for test concentrations: ca. 3

- Range finding study: not reported

Reference substance (positive control):

yes

Remarks:

Antimycin A

Key result

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

250 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Remarks:

active acid

Basis for effect:

mortality (fish)

Remarks on result:

other: (220-280)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

250 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Remarks:

active acid

Basis for effect:

mortality (fish)

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

114 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Remarks:

activ acid

Basis for effect:

mortality (fish)

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

35 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Remarks:

active acid

Basis for effect:

behaviour

Details on results:

- Observations on body length and weight: weight measurements of surviving fish at the end of the study yielded the following weight percentages of the control group mean weight: 35 mg/l, 128%,; 62 mg/l, 114%; 114 mg/l, 112% and 205 mg/l 97%.

- Other biological observations: Although no mortalities were observed at 114 mg/l, all test fish at this concentration exhibited loss of equilibrium within 30 minutes after beginning of the test. All test fish at this concentration recovered from this apparent initial toxicant shock, the same behavioral response was noted immediately following each solution change at 114, 205 and 345 mg/l.

- Mortality of control: 0%

Results with reference substance (positive control):

- Mortality: 100% mortality at 0.0014 mg/l

- LC50: 0.000084 mg/l

Reported statistics and error estimates:

Litchfield and Wilcoxon

Sublethal observations / clinical signs:

Table 1. Percentage mortalities

Mean measured concentration    (mg active acid/L)	Days of exposure
	1	2	3	4	5	6	7	8	9	10	11	12	13	14
Control	0	0	0	0	0	0	0	0	0	0	0	0	0	0
35	0	0	0	0	0	0	0	0	0	0	0	0	0	0
62	0	0	0	0	0	0	0	0	0	0	0	0	0	0
114	0	0	0	0	0	0	0	0	0	0	0	0	0	0
201	17	17	17	17	17	17	17	17	17	17	17	17	17	17
350	97	97	97	97	97	97	97	97	97	97	97	97	97	97

Validity criteria fulfilled:

yes

Conclusions:

A 14 d and 96 h LC50 value of 250 mg active acid/l and a NOEC value of 114 mg/l have been determined for the effects of the test substance on the mortality of the freshwater fish O. mykiss based on mean measured concentrations.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The study was conducted according to an appropriate OECD test guideline. It was not compliant with GLP and no analytical monitoring was carried out.

Qualifier:

according to guideline

Guideline:

other: "Methods for acute toxicity tests with fish, macroinvertebrates and amphibians" EPA-660/3-75-009 (US EPA)

Deviations:

not applicable

Remarks:

guideline not available

GLP compliance:

no

Specific details on test material used for the study:

Test material neutralised to pH 7.0 with NaOH.

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: an appropriate amount of test substance was added directly to the water in each vessel to obtain the desired concentration

- Controls: sea water only

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM

- Common name: sheepshead minnow
- Source: collected from Big Lagoon, near Bionomics Marine Research Laboratory (BMRL)
- Length at study initiation (length definition, mean, range and SD): 15-20 mm
- Feeding during test: no


ACCLIMATION

- Acclimation period: 7 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: not reported
- Feeding frequency: not reported
- Health during acclimation (any mortality observed): <1% mortality

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20±1⁰C

pH:

initial pH range in exposure media: 5.7 to 6.9
Although the pH in highest test concentration was below the validity criterion, 5.8 to 5.9, it is not thought by the reviewer that this would have significantly affected the outcome of the study.
pH in controls: 8.2 to 8.3

Dissolved oxygen:

≥60%

Salinity:

25‰

Nominal and measured concentrations:

Nominal concentrations: control, 1800, 3200, 5600, 10 000 and 18 000 µ/l (ppm)

Details on test conditions:

TEST SYSTEM

- Test vessel: jars
- Type (delete if not applicable): open
- Material, size, fill volume: 19 L glass jars, filled with 15L
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: natural, filtered sea water
- Culture medium different from test medium: no
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: not reported
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortalities were recorded every 24 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 1.8
- Range finding study: none reported

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

8 000 other: (µl/l) ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: equivalent to 1600 mg active acid/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

9 960 other: µl/l (ppm)

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: equivalent to 2000 mg active acid/L

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

9 960 other: µl/l (ppm)

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: equivalent to 2000 mg active acid/L

Reported statistics and error estimates:

LC50s were calculated by linear regression

Sublethal observations / clinical signs:

Table 1. Summary of mortalities.

Nominal concentrations ppm (µl/l)	Mortality (%)
	24 h	48 h	96 h
Control	0	0	0
1800	0	0	10
3200	0	10	10
5600	0	0	10
10 000	30	30	40
18 000	100	100	100

The concentrations have been reported in terms of µl/l (ppm) and whole test substance. The test substance has a density of 1.21 and an active acid content of 16.3%. Therefore the results in terms of active acid are:

24 h LC50 9960 ppm (*1.21*16.3%) = 2000 mg active acid/L

48 h LC50 9960 ppm (*1.21*16.3%) = 2000 mg active acid/L

96 h LC50 8000 ppm (*1.21*16.3%) = 1600 mg active acid/L

Validity criteria fulfilled:

yes

Conclusions:

A 96 h LC50 value of 8000 µl/l (ppm), equivalent to 1600 mg active acid/L has been reported for the effects of the test substance on the mortality of the marine fish C. variegatus.

Description of key information

14 d and 96 h LC₅₀: 250 mg active acid/l and NOEC: 114 mg active acid/l, O. mykiss, reliability 1 (ABC 1979).

96 h LC₅₀: 1600 mg active acid/l, C. variegatus, reliability 2 (EG&G 1976).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

250 mg/L

Marine water fish

Marine water fish

Effect concentration:

1 600 mg/L

Additional information

A 14 d and 96 h LC₅₀ value of 250 mg active acid/l and NOEC value of 114 mg/l have been determined for the effects of the test substance on the mortality of the freshwater fish O. mykiss based on mean measured concentrations (ABC, 1979). This study has been selected as key as it is the lowest reliable LC₅₀ value with freshwater fish.

A 96 h LC₅₀ value of 8000 µl/l (ppm), equivalent to 1600 mg active acid/l has been reported for the effects of the test substance on the mortality of the marine fish C. variegatus (EG&G, 1976 (6.1.1.004)). This study has been selected as key as it is the lowest reliable LC₅₀ value for marine fish.

A supporting study has also reported a 96 h LC₅₀ value of 970 mg active acid/l determined for the effects of the test substance on the mortality of the marine fish Ictalurus punctatus, based on nominal concentrations (EG&G, 1976 (6.1.1.003)).