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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Qualifier:
according to
Guideline:
other: US EPA 1975
Deviations:
not applicable
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: an appropriate amount of test substance was added to each vessel and dispersed

- Controls: sea water only
Test organisms (species):
Palaemonetes pugio
Details on test organisms:
TEST ORGANISM

- Common name: grass shrimp

- Source: collected from Big Lagoon, near Bionomics Marine Research Laboratory.

- Length at study initiation (range): 15-20 mm


- Method of breeding: not reported

- Feeding during test: no


ACCLIMATION

- Acclimation period: min. 7 days

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

- Type and amount of food: 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:
21 ± 1⁰C
pH:
Control: 8.1 to 8.3, exposure concentrations: 6.2 to 7.2. The pH decreased with time in the control, and increased with time in the exposure vessels.
Dissolved oxygen:
≥60%
Salinity:
25‰
Nominal and measured concentrations:
Nominal concentrations: Control, 1000, 1800, 3200, 5600, 10 000 µl/L (ppm)
Details on test conditions:
TEST SYSTEM

- Test vessel: jars

- Type (delete if not applicable): open

- Material, size, fill volume: glass, 19 L which contained 15L of seawater

- 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 (5 micrometers) sea water

- Intervals of water quality measurement: 24 hours


OTHER TEST CONDITIONS

- Adjustment of pH: none

- 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: 1.8 to 2

- Range finding study: not reported to have been conducted
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
800 other: µl/l (ppm)
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
acitve acid
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
3 160 other: µl/l (ppm)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: (1720 - 5810)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
3 300 other: µl/l (ppm)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: (1790-6075)
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
3 300 other: µl/l (ppm)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: (1790-6075)
Details on results:
- Mortality of control: 0%
Reported statistics and error estimates:
LC50s were determined by regression analysis.

Table 1. Summary of findings.

 Nominal concentration (u/l; ppm)  Mortality (%)
 24 h  48 h  96 h
 Control  0  0  0
 1000  0  0  0
 1800  0  0  0
 3200  10  10  30
 5600  100  100 100 
 10 000  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.

A 96 h LC50 value of 3160 µl/l (ppm) of test substance has been determined. This is equivalent to 800 µl/l (ppm) in terms of HMDTMP active acid. The specific gravity of the test substance is 1.23 therefore the results in terms of mg/l are:

HMDTMP-xK = 96 h LC50 3900 mg/l

HMDMTP acid = 96 h 1000 mg/l

Validity criteria fulfilled:
yes
Conclusions:
A 96 h LC50 value of 3900 mg/l (3160 ppm) of test substance, corresponding to 1000 mg/l active acid, has been determined for the effects of the test substance on the mortality of the marine amphipo Palaeomonetes pugio based on nominal concentrations.
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Qualifier:
according to
Guideline:
other: In house protocol: MIC Environmental Assessment Method for Conducting Acute Toxicity Tests with Daphnia magna (see note below)
Deviations:
no
Qualifier:
according to
Guideline:
other: US EPA (1975) Methods for acute toxicity tests with fish, macroinvertebrates and amphibians. EPA 660/3-75-009.
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
not applicable
Principles of method if other than guideline:
The In house protocol references Grueber and Adams (1980)
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: For each test concentration the appropriate amount of the test compound was pipetted into 1L of dilution water and shaken vigorously for 1 min. The solution was then divided into 2 200 ml aliquots in triplicate beakers to provide the appropriate replication. The remaining 400 ml were used for 0 h DO, pH, alkalinity and hardness determinations.

- Controls: Just dilution water.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM

- Common name: water flea

- Source:  The daphnids used in the test were obtained from a laboratory culture at the MIC laboratory.

- Age at study initiation (mean and range, SD): <24 h

- Method of breeding: according to lab SOP

- Feeding during test: no


ACCLIMATION

- Acclimation period: not reported

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

- Type and amount of food: Daphnia were fed a mixture of Purina trout chow and alfalfa daily.

- Health during acclimation (any mortality observed): not reported
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
The alkalinity ranged from 236 to 304 mg/L and 250 to 300 mg/L in the low and high test concentration vessels respectively.
Test temperature:
Average temperature of 21.2 degrees C.
pH:
The pH in the control medium ranged between 7.2-8.4. The pH in the test media ranged between 2.0 and 8.2.
Dissolved oxygen:
The dissolved concentrations for the treatments were not reported due to measurement interference from the Dequest products. However, DO on the control water ranged from 6.2-9.4 mg/L. DO was measured by the Winkler titriation method.
Nominal and measured concentrations:
Nominal test concentrations were 125, 250, 500, 1000 and 2000 mg/l as active acid
Details on test conditions:
TEST SYSTEM

- Test vessel: beaker

- Material, size, headspace, fill volume: 250 ml beakers containing 200 ml of 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 from St. Peters, Missouri.

- Alkalinity: 303 mg/L CaCO3, hardness 297 mg/L CaCO3

- Culture medium different from test medium: not reported

- Intervals of water quality measurement:  DO, pH, alkalinity, hardness and temperature of test solutions were monitored at the initiation (control only) and termination of the test in the high, middle and low concentrations. 


OTHER TEST CONDITIONS

- Adjustment of pH: no

- Photoperiod: not reported

- Light intensity: not reported


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility at 0, 24 and 48 h.


TEST CONCENTRATIONS

- Spacing factor for test concentrations: 2

- Range finding study

- Test concentrations: not reported

- Results used to determine the conditions for the definitive study: EC50 833 mg/L
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
125 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: immobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
570 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: immobility
Remarks on result:
other: 95% CL 500-660
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
680 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: immobility
Remarks on result:
other: 95% Cl 500-1000
Details on results:
- Mortality of control: 0%
Reported statistics and error estimates:
EC50 values and 95% confidence intervals calculated using an LC50 computer programme developed by Stephen et al (1978).   

Table 1. Summary of findings (average).

 Nominal concentrations (mg/L)     Percent immobilisation 
24 h 48 h 
Control 
125  6.7 
250  3.3 10.0
500  3.3 16.7
1000  100  100 
2000  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.

The pH data suggest that the acidity of the higher concentration test solutions is likely to have contributed to the effects observed in the test.


Validity criteria fulfilled:
no
Remarks:
pH very low at the highest concentrations tested. However the reported EC50 value is high enough to determine toxicity that the test substance is of low intrinsic toxicity.
Conclusions:
A 48 h EC50 value of 570 mg active acid/L has been determined for the effects of HMDTMP acid on the immobility of the freshwater flea D. magna. The results indicate that the substance is of very low short-term toxicity to invertebrates. However, they should be treated with caution the mortalities may be an effect of the pH of the test medium, rather than a reflection of true toxicity.
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See attachment for justification of read-across
Reason / purpose:
read-across source
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
570 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
active acid
Basis for effect:
other: immobility

Description of key information

48 h EC50 570 mg active acid/L, D. magna, reliability 2 (Monsanto 1981, read-across from HMDTMP acid CAS 23605-74-5)

96 h LC50 1000 mg active acid/L, P. pugio, reliability 2 (EG&G 1976)

96 h EC50 210 mg active acid/L, C. virginica, reliability 2 (EG&G 1977)

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
570 mg/L
EC50/LC50 for marine water invertebrates:
1 000 mg/L

Additional information

A 48 h EC50 value of 570 mg active acid/L has been determined for the effects of HMDTMP acid on the immobility of the freshwater flea Daphnia magna based on nominal concentrations. The pH in the test media ranged from 2.0 to 8.2, however the results are consistent with expectations based on data available with analogue substances. Therefore, while results indicate that the substance is of low short-term toxicity to invertebrates, they should be treated with caution because the mortalities may be an effect of the pH of the test medium, rather than a reflection of true toxicity.

A 96 h LC50 value of 3900 mg/l HMDTMP K pH 7, corresponding to 1000 mg/l active acid, has been determined for the effects of the test substance on the mortality of the marine amphipod Palaeomonetes pugio based on nominal concentrations.

Supporting data are available with the registered substance indicating a low short-term toxicity. The results ranged from 555 to 4660 mg active acid/L with D. magna.

A sub-lethal study measuring the shell deposition growth with the marine eastern oyster Crassostrea virginica and HMDTMP K pH 7 reported a 96 h EC50 value of 210 mg active acid/L (EG&G 1977). Shell growth is dependant on calcium bioavailability, which is greatly reduced by phosphonates due to their ability to form strong metal complexes. Therefore the outcome of this study is negatively affected by the substance forming complexes with calcium and the results should not be interpreted as inherent toxicity effects.