[nitrilotris(methylene)]trisphosphonic acid, ammonium salt

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2003-11-28 to 2003-12-19

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The result was obtained by valid read-across.

Qualifier:

according to guideline

Guideline:

other: PARCOM 1995 part B

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

no range finding test, no determination of the test substance in solution

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM

- Common name: sheepshead minnow

- Source: Aquatic Research Organisms


ACCLIMATION

- Acclimation period: 13.11.03 to 19.12.03

- Acclimation conditions (same as test or not): synthetic seawater, 17.6-22.4 degC, salinity 33%o, DO >60%, flow-through

- Feeding frequency: at least once per working day

- Health during acclimation (any mortality observed): 0%

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

Not reported

Test temperature:

19 +/- 1 degC

pH:

range: 8.1-8.2

Dissolved oxygen:

range: 95-100%

Salinity:

range: 3.2-3.5 %o

Nominal and measured concentrations:

Nominal concentrations: 0, 100, 300 and 1000 mg/L

Details on test conditions:

TEST SYSTEM

- Test vessel: not reported, assumed to be according to guideline

- Renewal rate of test solution (frequency/flow rate): not reported, assumed to be according to guideline

- 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: reference sea water was bought from Bjolsen Akvarium AS

- Intervals of water quality measurement: pH, DO, salinity and temperature were measured daily in all treatments.



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at 24, 48, 72 and 96 h


TEST CONCENTRATIONS

- Spacing factor for test concentrations: 3

- Justification for using less concentrations than requested by guideline: none given

- Range finding study

- Results used to determine the conditions for the definitive study: range finding test with S. costatum, test concentrations based on findings, no further details reported.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol (2.3 mg/L)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 400 - 500 mg/L

Nominal / measured:

nominal

Conc. based on:

other: ATMP 5Na salt

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 282 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

Results with reference substance (positive control):

- Results with reference substance valid? yes, valid if mortality 20-80%

- Mortality: 20% after 96 h

Reported statistics and error estimates:

LC50 and 95% confidence intervals were calculated using a Logistic Regression Model.

Sublethal observations / clinical signs:

After 24 h 1 individual died in the highest treatment. This was the only death recorded.

Result expressed as nominal concentration. Properties of the test substance and evidence from other studies (where concentrations were 

measured) indicate that nominal and measured concentrations are likely to be in good agreement.

The study reports results as whole substance tested. However for the purpose of the risk characterisation conversion of results into acid and salt of ATMP equivalent was required. This was achieved by using the reported active acid content of the substance (28.2%) and the known ATMP 5 Na salt content of the test substance (ca. 40 -50%).

Therefore:

test substance 96 h LC50 = >1000 mg/L

ATMP acid 96 h LC50 = >282 mg/L (1000*28.2%)

ATMP 5Na salt 96 h LC50 = >400 -500 mg/L [1000*(40%-50%)]

Validity criteria fulfilled:

yes

Conclusions:

A 96 h LC50 value of >400-500 mg/L has been determined for the effects of ATMP 5Na salt on the mortality of the marine fish Cyprinodon variegatus. A 96 h LC50 value of >282 mg/L has also been determined for the effects of ATMP as active acid on mortality of C. variegatus.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1979-04-11 to 1979-04-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to a test protocol that is comparable to the appropriate OECD test guideline, and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

other: APHA (1975) Standard methods for the examination of water and wastewater. 14th edition. US EPA (1975) Methods for acute toxicity test with fish, macroinvertebrates and amphibians (EPA-660/3-75-009).

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

Deviations:

yes

Remarks:

no range finding test was conducted

Principles of method if other than guideline:

Method: other

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all test concentration

- Sampling method: concentrations were determined on days 0, 1, 5, 10 and 14.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)

- Method: mechanically dissolved in deionised water

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM

- Common name: rainbow trout

- Source: Test fish were obtained from Spring Creek Hatchery, Lewiston, Montana.

- Length at study initiation (length definition, mean, range and SD): initial mean standard length of 33 mm.

- Weight at study initiation (mean and range, SD): initial mean weight of 0.64 g

- Feeding during test: no data


ACCLIMATION

- Acclimation period: Acclimated to test conditions for at least 14 days.

- Acclimation conditions (same as test or not): Fish culture techniques were those described in Brauhn et al. (1975)

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

14 d

Post exposure observation period:

none reported

Hardness:

240 mg/L as CaCO3

Test temperature:

12 +/- 1 degree C. 

pH:

The pH of the test media ranged between 6.3 in the high concentration to 7.9 in the control.

Dissolved oxygen:

>7.5 mg/L

Salinity:

Not Applicable

Nominal and measured concentrations:

Nominal test concentrations were 19, 42, 80, 163 and 320 mg/L (active acid). 
Mean measured concentrations were 19, 47, 94, 196 and 390 mg/L (active acid).

Details on test conditions:

TEST SYSTEM

- Test vessel:

- Material, size, headspace, fill volume: 30L glass test aquaria

- Aeration: aerated well water

- Type of flow-through (e.g. peristaltic or proportional diluter): Mount & Brung proportional diluter

- Renewal rate of test solution (frequency/flow rate): replacement rate of 200 ml/min/test vessel. 

- No. of organisms per vessel: 30

- No. of vessels per concentration (replicates): 1

- No. of vessels per control (replicates): 1



TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: ABC laboratories well water

- Metals: all <0.1 mg/L

- Alkalinity: 360 mg/L as CaCO3

- Culture medium different from test medium: no

- Intervals of water quality measurement: Temperature, DO, and ammonia were measured on days 0, 1, 5, 10, and 14 in control, low concentration, and high concentration samples. The values were all acceptable.



OTHER TEST CONDITIONS

- Photoperiod: 16h day light



EFFECT PARAMETERS MEASURED (with observation intervals if applicable): weight of fish were measured at the beginning and the end of the test, mortality and abnormal behaviour were checked every 24 h.



TEST CONCENTRATIONS

- Spacing factor for test concentrations: 2

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

160 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: (95% CL 130-190)

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

47 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

150 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: Although only 10% mortality was observed at 94 mg/l, all test fish at this concentration exhibited partial loss of equilibrium within 30 minutes after beginning the test. As the test was extended, this group recovered from the apparent initial shock of toxicant exposure and responded normally for the duration of the test.

- Observations on body length and weight: the study yielded the following weight percentages of the control group mean weight: 19mg/L - 113%, 47 mg/L - 107%, 94 mg/L - 93% and 196 mg/L - 63%. The average weight of the control group at the end of the test showed a 59% increase in the growth when compared with the representative group of fish at the beginning of the test.

- Mortality of control: 0%

Reported statistics and error estimates:

LC50 and 95% confidence intervals determined by the method of Litchfield and Wilcoxon.

Sublethal observations / clinical signs:

Result expressed as measured concentration

24 - 72 hr LC50: 180 mg (active acid)/L
Day 4 - Day 8 LC50: 160 mg (active acid)/L
Day 9 - Day 14 LC50: 150 mg (active acid)/L

Validity criteria fulfilled:

no

Conclusions:

A 96 h LC50 value of 160 mg/L (as active acid) has been determined for the effects of the test substance on mortality of S. gairdneri (now known as Onychorhynchus mykiss). Also a 14 day NOEC of 47 mg active acid/L and a 14 day LC50 of 150 mg active acid/L have been determined for the effects of the test substance on mortality of S. gairdneri (now known as O. mykiss).

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Please refer to Annex 3 of the CSR and IUCLID Section 13 for justification of read-across within the ATMP category.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

160 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: (95% CL 130-190) (ABC, 1979)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 400 - 500 mg/L

Nominal / measured:

nominal

Conc. based on:

other: ATMP 5Na salt

Basis for effect:

mortality (fish)

Remarks on result:

other: AnalyCen (2003)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 282 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: AnalyCen (2003)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: AnalyCen (2003)

Reference 4

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

The study was assigned a reliability score of 1 in a thorough, peer-reviewed report by the Environment Agency (2007). However, the study has been assigned a reliability value of 2, as there is not enough information on control mortality to compare to validity criteria from modern guidelines.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The sensitivity of early life stages of Oncorhynchus mykiss to ammonia was tested in 96-hour exposures. Emergence, survival and yolk absorption of fish were measured and the total ammonia concentrations were measured using an automated method that measured the intensity of indophenol blue, formed after the reaction of ammonnia with alkaline phenol hypochlorite.

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

Water samples were taken and the total ammonia concentrations were analyzed on the same day. Further information on sampling was not reported.

Details on test solutions:

No information on the preparation of test solutions was reported.

Test organisms (species):

Oncorhynchus gorbuscha

Details on test organisms:

TEST ORGANISM
- Common name: pink salmon
- Source: wild caught from Lovers Cove Creek, Southern Baranof Island, Alaska.
- Strain: not reported
- Method of breeding: fertilised eggs were collected from Lovers Cove Creek, Southern Baranof Island, Alaska.


ACCLIMATION - not reported except that the eggs were held in Heath incubators.

QUARANTINE (wild caught) - no information reported

FEEDING DURING TEST (as applicable) - no information reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Water hardness was not reported

Test temperature:

3.7 - 4.8 degrees C

pH:

6.3 - 6.5

Dissolved oxygen:

Dissolved oxygen not reported

Salinity:

Salinity not reported

Conductivity:

Conductivity not reported

Nominal and measured concentrations:

Nominal: values not reported
Measured: lowest test concentration: 50 ppb NH3, highest test concentration: >1500 ppb NH3.

Details on test conditions:

TEST SYSTEM
- Test vessel: 18 L test vessel
- Type (delete if not applicable): not reported
- Aeration: test solutions were aerated, further information not reported
- No. of organisms per vessel: 25
- No. of vessels per concentration (replicates): not reported
- No. of vessels per control (replicates): not reported
- Biomass loading rate: <0.3 g tissue/L test solution

TEST MEDIUM / WATER PARAMETERS - not reported

OTHER TEST CONDITIONS - not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : emergence, survival and absorption of the yolk

TEST CONCENTRATIONS - not reported

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.068 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: results expressed in terms of mg NH3-N/L

Details on results:

- Other abnormalities: not reported
- Other biological observations: Hatching of eyed eggs was not affected by concentrations >1500 ppb. The most sensitive life stage was late alevins, determined using the median tolerance limit (TLM).

Reported statistics and error estimates:

Median tolerance limits (TLM) and 95% confidence levels were calculated by Probit analysis (Finney, 1971).

Validity criteria fulfilled:

not applicable

Remarks:

No guideline followed. Not enough detail on raw data to directly compare to validity criteria from modern test guidelines.

Conclusions:

A 96-hour LC50 value of 0.068 mg NH3-N/L was reported for the effects of ammonia on Oncorhynchus gorbuscha.

Description of key information

There are no test data for ATMP-xNH₄, however data have been read-across from ATMP-H and ATMP-xNa.

96-hour LC₅₀ 160 mg active acid/L Salmo gairdneri (now known as Oncorhynchus mykiss), read-across from ATMP-H

96-hour LC₅₀ >282 mg active acid/L marine fish Cyprinodon variegatus, read-across from ATMP-xNa

Ammonia: 96-hour LC₅₀ 0.068 mg NH₃-N/L, Oncorhynchus gorbuscha

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

160 mg/L

Fresh water fish

Dose descriptor:

LC50

Remarks:

toxicity data for ammonia

Effect concentration:

0.068 mg/L

Marine water fish

Marine water fish

Dose descriptor:

LC50

Effect concentration:

> 282 mg/L

Additional information

ATMP-xNH₄ will dissociate when it is released into aqueous environments to yield ATMP and ammonia. No data is available for ATMP ammonium salts, therefore data are read-across from ATMP-H and ATMP-xNa.

In a reliable, 96-hour exposure to Oncorhynchus mykiss, an LC₅₀ value of 160 mg active acid/L was determined for the effects of ATMP-H on mortality (ABC, 1979). The study was conducted as a prolonged toxicity to fish study, and so 14-day LC₅₀ and NOEC values of 150 and 47 mg active acid/L were also determined for the mortality of O. mykiss. This study reflects the lowest freshwater values that are available for this endpoint.

A reliable study has been read-across from ATMP-xNa which gives a 96-hour LC₅₀ value of >282 mg active acid/L for the mortality of the marine fish, Cyprinodon variegatus (AnalyCen, 2003). This value represents the lowest value available for the marine compartment for this endpoint.

Several other supporting studies are available:

Schoberl & Huber (1988) report an LC₅₀ value of 240 mg/L for the toxicity of ATMP-H to the mortality of Cyprinus sp. however, little information on the study methodology was reported and so the study was assigned a Reliability score of 4.

A 48-hour LC₅₀ value of >250 mg active acid/L has been reported for the effects of ATMP-H to the mortality of Ides fish species however, only a summary report was available for review and the study was assigned Reliability 4 accordingly (Henkel, 1972).

A secondary literature source reported 48-hour LC₅₀ and NOEC values of 250 and 150 mg/L for the toxicity of ATMP-H on the mortality of Leuciscus idus (Kastner & Gode, 1983).

Two reliable EG&G studies were conducted. The first reported a 96-hour LC₅₀ of 1212 mg active acid/L for the effects of ATMP-H on the mortality of Ictalurus punctatus, however <60% oxygen saturation level was recorded at the end of the test (EG&G, 1976b). The second study reported a NOEC of 4831 mg active acid/L for the effects of ATMP-H on Cyprinodon variegatus (EG&G, 1976c).

Lastly, a 48-hour LC₅₀ value of 1946 mg active acid/L has been reported in a reliable study for the effect of ATMP-4Na on the mortality of Salmo gairdneri (new name: Oncorhynchus mykiss) (Inveresk, 1982).

Ammonia: A 96-hour LC₅₀ value of 0.068 mg NH₃-N/L for the effects of unionised ammonia on Oncorhynchus gorbuscha (Rice and Bailey, 1980). The value is equivalent to 0.083 mg NH₃/L. The study was reviewed in the report: Environment Agency Proposed EQS for Water Framework Directive Annex VIII Substances: Ammonia (unionised) 2007 (EA, 2007).

ATMP-xNH₄ will dissociate as it is released into the aquatic environment and it is therefore necessary to assess the hazard properties and derive PNECs for ATMP and ammonia separately.

For hazard classification, ATMP-xNH₄ has been assessed based on the available aquatic data for the whole substance.

The influence of the ammonium salt is discussed in IUCLID Section 6.0 Ecotoxicological Information and Section 7.0 of the CSR.

The acid, sodium, potassium and ammonium salts in the ATMP category are freely soluble in water. The ATMP anion can be considered fully dissociated from its sodium, potassium or ammonium cations when in dilute solution. Under any given conditions, the degree of ionisation of the ATMP species is determined by the pH of the solution. At a specific pH, the degree of ionisation is the same regardless of whether the starting material was ATMP-H, ATMP.4Na, ATMP.7K or another salt of ATMP.

 

Therefore, when a salt of ATMP is introduced into test media or the environment, the following is present (separately):

1. ATMP is present as ATMP-H or one of its ionised forms. The degree of ionisation depends upon the pH of the media and not whether ATMP (3-5K) salt, ATMP (3-5Na) salt, ATMP-H (acid form), or another salt was used for dosing.
2. Disassociated potassium, sodium or ammonium cations. The amount of potassium or sodium present depends on which salt was dosed.
3. It should also be noted that divalent and trivalent cations would preferentially replace the sodium or potassium ions. These would include calcium (Ca²⁺), magnesium (Mg²⁺) and iron (Fe³⁺). These cations are more strongly bound by ATMP than potassium, sodium and ammonium. This could result in ATMP-dication (e.g. ATMP-Ca, ATMP-Mg) and ATMP-trication (e.g. ATMP-Fe) complexes being present in solution.

In this context, for the purpose of this assessment, read-across of data within the ATMP Category is considered to be valid.