Reaction product of a ((2S)-2, 6-diaminohexanoic acid hydrochloride salt, phosphonic acid and methanal) with a 1,2-disubstituted alkane and sodium hydroxide.

Short-term toxicity to fish

Administrative data

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:

1979-04-11 to 1979-04-25

Reliability:

2 (reliable with restrictions)

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 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:

other: The study was conducted according to an appropriate national standard test method. 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, USEPA, 1975

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not applicable

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: the test substance was added directly into the each jar and mixed by stirring

- Controls: dilution water

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM

- Common name: rainbow trout

- Source: Fish were obtained from a commercial fish hatchery in Washington, USA

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

- Weight at study initiation (mean and range, SD): mean wet weight of 1.9 grams at time of study

- Method of breeding: not reported

- Feeding during test: no


ACCLIMATION

- Acclimation period: 48 h

- Acclimation conditions (same as test or not): same as test

- Type and amount of food: animals were not fed during acclimation period

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

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

35 mg/L CaCO3

Test temperature:

Water temperature was maintained at 12 deg C.

pH:

The pH of the test media was not reported. However a test was also conducted with Icatlurus punctatus in similar test conditions (higher temperature) and higher test concentrations, which reported a pH range of 7.0-8.2.

Dissolved oxygen:

50-99% saturation. The DO is below the recommended OECD level of 60%.

Nominal and measured concentrations:

Nominal test concentrations were 576, 432, 324, 252, and 180 mg/l as active acid. 

Details on test conditions:

TEST SYSTEM

- Test vessel: jars

- Material, size, headspace, fill volume: 19 L glass jars filled with 15L test medium

- Aeration: no

- No. of organisms per vessel: 10

- No. of vessels per concentration (replicates): 3

- No. of vessels per control (replicates): 3


TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: well water

- Culture medium different from test medium: same

- Intervals of water quality measurement: DO and temperature were measured at 0, 48 and 96 hours in the highest, middle and lowest test concentration exposures and found to be acceptable. 


OTHER TEST CONDITIONS

- Adjustment of pH: not reported

- Photoperiod: not reported

- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality every 24 h


TEST CONCENTRATIONS

- Spacing factor for test concentrations: 1.35

- Range finding study: yes, details not provided

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

216 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: estimated, between 180 and 252 mg/l concentrations

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

- Other: no mortalities were recorded at 180 mg/l, while 100% mortality was recorded at 252 mg/l concentration. Threfore the 96 h EC50 value of 215 mg active acid/l is an estimated EC50.

Reported statistics and error estimates:

LC50 values and 95% confidence intervals determined by regression analysis of concentrations expressed as logarithms and mortality expressed as probits.

Sublethal observations / clinical signs:

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.

Percentage mortality at each nominal test concentration:

Concentration (mg/L)	Mortality at 96 h (%)
Control	0
180	0
252	100
324	100
432	100
576	100

24 hr LC50: 252 (203-313) mg/L 
48 hr LC50: >180<252 mg/L 96 hr LC50: >180<252 mg/L

Validity criteria fulfilled:

no

Remarks:

DO was low

Conclusions:

A reliable 96 h LC50 value of 180-252 mg/L, i.e. 216 mg/l has been determined for the effects of the test substance on the mortality of the freshwater fish Salmo gairdneri (new name: Oncorhynchus mykiss). The results should however be treated with caution as the DO levels decreased to 50%, while the concentrations at which this occurred are not reported. The result is considered valid as the controls mortality was 0%.

Reference 3

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:

other: The study was conducted according to an appropriate EU test method. 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, USEPA, 1975

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

not applicable

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: appropriate amounts of test substance were directly added to the vessels

- Controls: dilution water only

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM

- Common name: sheepshead minnow

- Source: collected from stocked ponds at the testing laboratory 

- Length at study initiation (length definition, mean, range and SD): 15-20 mm standard length

- Method of breeding: not reported


ACCLIMATION

- Acclimation period: a minimum of 7 days before testing

- Acclimation conditions (same as test or not): not reported (only "acclimation to laboratory conditions" reported)

- Type and amount of food: not reported

- Health during acclimation (any mortality observed): <1%

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Water temperature was maintained at 20 +/- 1 degree C.

pH:

The pH of the test media ranged between 6.7 and 8.3

Dissolved oxygen:

>= 58% saturation for all concentrations and control

Salinity:

25 parts per thousand

Nominal and measured concentrations:

Nominal test concentrations were 1000, 10000, 18000, 32000 and 56000 ml/l (ppm).

Details on test conditions:

TEST SYSTEM

- Test vessel: jars

- Type (delete if not applicable): open

- Material, size, headspace, fill volume: Testing was conducted in 19 litre uncovered glass jars containing 15 litres of test medium.

- 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: Filtered natural sea water.

- Intervals of water quality measurement: DO and pH were measured during the study and remained within acceptable limits


OTHER TEST CONDITIONS

- Adjustment of pH: not reported

- Photoperiod: not reported

- Light intensity: not reported


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, every 24 h


TEST CONCENTRATIONS

- Spacing factor for test concentrations: 2 and 10

- Range finding study: not reported

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6 435 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Reported statistics and error estimates:

Test concentrations were converted to logarithms and corresponding percentage mortalities to probits. LC50 values were then determined by linear regression.

Sublethal observations / clinical signs:

Table 1. Mortality of fish during 96 h exposure.

Nominal concentration (u/l; ppm)	Mortality (%)
	24 h	48 h	96 h
Control	0	0	0
1 000	0	0	0
10 000	0	0	0
18 000	0	10	10
32 000	60	60	60
56 000	100	100	100

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.

24 hr LC50: 5377 mg/l (30,299 ppm)

48 hr LC50:

5377 mg/l (30,299 ppm)

96 hr LC50: 5377 mg/l (30,299 ppm)

The results reported have been corrected for active acid content (18%) and density (1.18).

Validity criteria fulfilled:

yes

Conclusions:

A reliable 96 h LC50 value of 6435 mg active acid/l has been determined for the effects of DTPMP acid towards the mortality of the marine fish Cyprinodon variegatus.

Description of key information

There are no studies available conducted on SPE1415, however there are three reliable (Klimisch 2) short-term toxicity studies in fish conducted on the analogue substances ATMP and DTPMP. In addition there is one reliable (Klimisch 2) study on another confidential analogue substance. These data are all included in this dossier.

One reliable (Klimisch 2) short-term toxicity study for fish was available for ATMP (ABC Laboratories, 1979). The GLP-compliant prolonged toxicity study with rainbow trout (Onchorhynchus mykiss, formerly known as Salmo gairdneri) provided a 96-hour LC50 of 160 mg/L and a 14-day LC50 of 150 mg/L, based on mean measured test concentrations of the ATMP active acid.

Two reliable (Klimisch 2) short-term toxicity studies for fish are available for DTPMP (EG&G, 1976a and b) and follow published study guidelines. The study with rainbow trout (Onchorhynchus mykiss, formerly known as Salmo gairdneri) provided a 96-hour LC50 of 216 mg/L based on nominal test concentrations of the active acid of DTPMP and the study with sheepshead minnow (Cyprinodon variegatus) provided a 96-hour LC50 of 5,377 mg/L based on nominal test concentrations of DTPMP active acid.

Additional supporting data on another confidential analogue substance is discussed in Additional Information.

Adequate reliable measured data exists for the short-term toxicity to fish to aminomethylphosphonate analogues of SPE1415 (ATMP and DTPMP).  In a conservative approach the most sensitive study result from these studies has been identified and used to address the hazard endpoint in question. The most sensitive study result has been identified as a reliable study with ATMP (ABC Laboratories, 1979) which reports an LC50 for short-term toxicity in fish of 150 mg/l. Consequently this value will be used for the short term toxicity to fish for SPE1415.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

150 mg/L