Registration Dossier

Diss Factsheets

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
24th January 1983- 28th January 1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
No CoA, no analytics, but all raw data included and QC,
Qualifier:
according to guideline
Guideline:
other: Standard Methods for the Examination of Water and Wastewater (1975). 14th edition
Qualifier:
according to guideline
Guideline:
other: Methods for Acute Toxicity Tests with Fish, Macroinverterbrates and Amphibians, (1975)
Version / remarks:
EPA 660/3/75-009
GLP compliance:
not specified
Remarks:
but quality control applied and reported
Specific details on test material used for the study:
Santicizer 8, NETSA, mixture of N-ethyl-o-toluene sulfonamide and N-ethyl-p-toluene sulfonamide, QM18
The Santicizer® 8 plasticizer test compound was received on January 7, 1983, in good condition. The sample upon receipt was observed to be a yellow liquid and was stored at 4°C. Sample purity was listed as >95%. Stability is recorded to be above 6 months and solubility in water as 13%. Test concentrations were prepared based on the total compound.
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
The test concentrations were obtained by transferring appropriate weights of the compound directly to the test chambers.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
The bluegill sunfish used in the test were obtained from Fattig Fish Hatchery in Brady, Nebraska. The fish were identified to species using the taxonomic keys developed by (Eddy,1969). All test fish were held in culture tanks on a 16 hour daylight photoperiod and observed for at least fourteen days prior to testing. Fish culture techniques used were basically those described by Brauhn & Schoettger(1975). A daily record of fish observations during the holding period, along with any therapeutic diseases treatments were recorded.

During this period, the fish received standard commercial fish food (Rangen’s) daily until 48 hours prior to testing at which time feeding was discontinued. The bluegills sunfish used for this experiment had a mean weight of 0.49g and a mean standard length of 26mm. The test vessels were kept in a water bath at 22°C (±1.0). The test fish were acclimated to the dilution water and test temperature and held without food for 48 hours prior to testing.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
255 ppm (CaCO3)
Test temperature:
start of the test: 23 °C
End of the test: 23 °C
pH:
Start of the test: 8
End of the test: 8-8.2
Dissolved oxygen:
Start of the test: 8.9-9.1 mg/L
End of the test: 3.8-6.1 mg/L
Conductivity:
50 µmhos/cm
Nominal and measured concentrations:
Nominal concentrations: 100, 180, 320, 560 and 1000 mg/L
Details on test conditions:
TEST SYSTEM
The static fish bioassay was conducted in five gallon glass vessels containing 15 liters of well water. A 72 hour range-finding test was conducted to determine the concentration range for the definitive study. The preliminary test concentration was set at 100 mg/l. Based on the results of preliminary testing, five concentrations of the test compound, ranging in a logarithmic series from 100 to 1000 mg/l, with ten fish per concentration were selected for definitive bioassay. The fish were added to the test chambers by random assignment within 30 minutes after addition of test material aliquots. Mortality and abnormal effects were monitored and recorded once every 24 hours.

TEST MEDIUM / WATER PARAMETERS
Parameter Concentration
Dissolved Oxygen 9.2 ppm
pH 7.8
Hardness (CaC03) 255 ppm
Alkalinity (CaC03) 368 ppm
Conductivity 50 µmhos/cm
Total Ammonia (NH3) <0 .10 ppm
N03-N 0.15 ppm
Ortho-Phosphate 0.10 ppm
Aluminum <0 .007 ppm
Arsenic <0 .001 ppm
Cadmium <0.001 ppm
Chromium <0 .002 ppm
Cobalt <0.002 ppm
Copper <0 .002 ppm
Iron <0 .004 ppm
Lead <0 .02 ppm
Mercury <0 .0001 ppm
Nickel <0 .004 ppm
Silver <0 .002 ppm
Zinc <0 .001 ppm

Measured organophosphorus pesticides: below detectable limits
Measured organochlorine pesticides plus PCB 's: below detectable limits




Reference substance (positive control):
yes
Remarks:
Antimycin A standard was obtained from Sigma Chemical Company, Type III, crystalline, Lot 89C-0588.
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
130 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% confidence limit ( 100-180)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
240 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% confidence limit (180-320)
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
420 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% confidence limit (320-560)
Details on results:
Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured throughout the test and were within acceptable limits outlined by the Committee on Methods for Toxicity Tests with Aquatic Organisms. Recorded dissolved oxygen ranged between 43% and 103% saturation at 22 degrees. It should be noted that under the used guideline, acceptable limits are considered to be above 40% whereas the OECD 203 guidelines specify a dissolved oxygen(DO) concentration of at least 60%. Although DO concentrations were above 60% in the controls, this was not the case for the other treatment groups after 96 hour, thus raising question to whether this had any impact on the final outcome. No mortality of fish in the control vessel were seen which fullfills a principle OECD validity criteria however there is no evidence that the concentration of the substance being tested has been satisfactorily maintained.
Results with reference substance (positive control):
The 96 hour LC50 for bluegill sunfish exposed to Antimycin A was 6.2Xl0^(-5) mg/l and was within the 95% confidence intervals reported in the literature (Berger et al, 1969).
Reported statistics and error estimates:
Statistical analysis of the concentration vs. effect data (generally mortality) was obtained by employing a computerized LC50 program developed by Stephan et al. (1977). This program calculated the LC50 statistic and its 95 percent confidence limits using the binomial, the moving average, and the probit tests. Three different methods of analyzing the data were used since not one method of analysis is appropriate for all possible sets of data that may be obtained. The method of calculation selected for use in this report was that which gives the narrowest confidence limits for the LC50 .
Sublethal observations / clinical signs:

Mortality rates during the acute toxicity test of Santicizer 8 Plasticizer to Bluegill Sunfish (Lepomis Macrochirus)

 

Percent mortality

Concentrations ( mg/L)

24 hr

48 hr

96 hr

Control

0

0

0

100

0

0

0

180

0

0

100

320

0

100

100

560

100

100

100

1000

100

100

100

Validity criteria fulfilled:
yes
Remarks:
see details on results
Conclusions:
The 96-h LC50 value is 130 mg/L (95% C.I 100-180 mg/L) for Bluegill Sunfish (Lepomis Macrochirus) based on nominal concentrations.
Executive summary:

Fish acute toxicity was examined in a EPA guideline study (EPA 660/3 -75 -009) study under quality control. In this study 10 bluegill sunfish (Lepomis macrochirus) per concentration, were exposed to nominal concentrations of 100, 180, 320, 560 and 1000 mg/L under static conditions for 96 hours. The 24 hour, 48 hour and 96 hour LC50 values for Santicizer 8 Plasticizer were determined as 420, 240 and 130 mg/L respectively and are based on nominal concentrations. Results indicated a 96 hour, no observed concentration of 100 mg/L based on mortality and abnormal effects. As a quality check, the bluegill sunfish were challenged with a reference compound, Antimycin A. The estimated 96 hour LC50 and 95% confidence limits (C.I.) were within the 95% confidence limits reported in the literature, indicating that the fish were in good condition.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24th January 1983 - 30th January 1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
No CoA, no analytics, but all raw data included and QC,
Qualifier:
according to guideline
Guideline:
other: Standard Methods for the examination of Water and Waste water (1975)
Qualifier:
according to guideline
Guideline:
other: Methods for Acute Toxicity Tests with Fish, Macroinverterbrates and Amphibians, (1975)
Version / remarks:
EPA 660/3/75-009
GLP compliance:
not specified
Remarks:
Quality assurance was applied and signed for.
Specific details on test material used for the study:
Santicizer 8, NETSA, mixture of N-ethyl-o-toluene sulfonamide and N-ethyl-p-toluene sulfonamide, QM18
The Santicizer® 8 plasticizer test compound was received on January 7, 1983, in good condition. The sample upon receipt was observed to be a yellow liquid and was stored at 4°C. Sample purity was listed as >95%. Stability is recorded to be above 6 months and solubility in water as 13%. Test concentrations were prepared based on the total compound.
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
Before the test concentrations were prepared, the standard was allowed to warm to room temperature. The test concentrations were obtained by transferring appropriate weights of the test compound directly to the test chambers.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
The rainbow trout used in the test were obtained from Trout Lodge in McMillin, Washington. The fishes were identified to species using the taxonomic keys developed by (Eddy,1969). All test fish were held in culture tanks on a 16 hour daylight photoperiod and observed for at least fourteen days prior to testing. Fish culture techniques used were basically those described by Brauhn & Schoettger(1975). A daily record of fish observations during the holding period, along with any therapeutic diseases treatments were recorded.
During this period, the fish received standard commercial fish food (Rangen’s) daily until 48 hours prior to testing at which time feeding was discontinued. The rainbow trout used for this experiment had a mean weight of 1.3 g and a mean standard length of 44mm. The test vessels were kept in a water bath at 12°C (±1.0). The test fish were acclimated to the dilution water and test temperature and held without food for 48 hours prior to testing.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
255 ppm (CaCO3)
Test temperature:
Start of the test: 12 °C
End of the test: 12 °C
pH:
Start of the test: 7.9-8.3
End of the test: 8-8.2
Dissolved oxygen:
Start of the test: 9.3-9.7
End of the test: 5.4-7.9
Conductivity:
50 µmhos/cm
Nominal and measured concentrations:
Nominal concentrations: 56, 100, 180, 320, 560 and 1000 mg/L
Details on test conditions:
TEST SYSTEM
The static fish bioassay was conducted in five gallon glass vessels containing 15 liters of well water. A 72 hour range-finding test was conducted to determine the concentration range for the definitive study. The preliminary test concentration was set at 100 mg/l. Based on the results of preliminary testing, six concentrations of the test compound, ranging in a logarithmic series from 56 to 1000 mg/l, with ten fish per concentration were selected for definitive bioassay. The fish were added to the test chambers by random assignment within 30 minutes after addition of test material aliquots. Mortality and abnormal effects were monitored and recorded once every 24 hours.

TEST MEDIUM / WATER PARAMETERS
Parameter Concentration
Dissolved Oxygen 9.2 ppm
pH 7.8
Hardness (CaC03) 255 ppm
Alkalinity (CaC03) 368 ppm
Conductivity 50 µmhos/cm
Total Ammonia (NH3) <0 .10 ppm
N03-N 0.15 ppm
Ortho-Phosphate 0.10 ppm
Aluminum <0 .007 ppm
Arsenic <0 .001 ppm
Cadmium <0.001 ppm
Chromium <0 .002 ppm
Cobalt <0.002 ppm
Copper <0 .002 ppm
Iron <0 .004 ppm
Lead <0 .02 ppm
Mercury <0 .0001 ppm
Nickel <0 .004 ppm
Silver <0 .002 ppm
Zinc <0 .001 ppm

Measured organophosphorus pesticides: below detectable limits
Measured organochlorine pesticides plus PCB 's: below detectable limits




Reference substance (positive control):
yes
Remarks:
Antimycin A standard obtained from Sigma Chemical Company, Type III, crystalline, Lot 89C-0588.
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
80 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% Cl (56-100)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
130 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% Cl (100-180)
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
240 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CL (180-320)
Details on results:
Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured throughout the test and were within acceptable limits outlined by the Committee on Methods for Toxicity Tests with Aquatic Organisms. Recorded dissolved oxygen ranged between 50% and 90% saturation at 12 degrees. However it should be noted that under this method, acceptable limits are considered to be above 40% whereas the OECD 203 guidelines specify a dissolved oxygen concentration of at least 60%. This deviation does not seem to have any impact on the final outcome. No mortality of fish in the control vessel were seen which fullfills a principle OECD validity criteria however there is no evidence that the concentration of the substance being tested has been satisfactorily maintained.
Results with reference substance (positive control):
The 96 hour LC50 for bluegill sunfish exposed to Antimycin A was 3.9Xl0^(-5) mg/l and was within the 95% confidence intervals reported in the literature (Berger et al, 1969).
Reported statistics and error estimates:
Statistical analysis of the concentration vs. effect data (generally mortality) was obtained by employing a computerized LC50 program developed by Stephan et al. (1977). This program calculated the LC50 statistic and its 95 percent confidence limits using the binomial, the moving average, and the probit tests. Three different methods of analyzing the data were used since not one method of analysis is appropriate for all possible sets of data that may be obtained. The method of calculation selected for use in this report was that which gives the narrowest confidence limits for the LC50 .
Sublethal observations / clinical signs:

Mortality rates during the acute toxicity test of Santicizer 8 Plasticizer to Rainbow Trout (Salmo Gairdneri)

 

Percent mortality

Concentrations ( mg/L)

24 hr

48 hr

96 hr

Control

0

0

0

56

0

0

0

100

0

10

90

180

0

100

100

320

100

100

100

560

100

100

100

 1000 100 100 100
Validity criteria fulfilled:
yes
Remarks:
see details on results
Conclusions:
The 96-h LC50 value is 80 mg/L (95% C.I 56-100 mg/L) for rainbow trout (Salmo Gairdneri) based on nominal concentrations .
Executive summary:

Fish acute toxicity was examined in a EPA guideline (EPA-660/3 -75 -009) study under quality control. In this study 10 rainbow trout (salmo gairdneri) per concentration, were exposed to nominal concentrations of 56, 100, 180, 320, 560 and 1000 mg/L under static conditions for 96 hours. The 24 hour, 48 hour and 96 hour LC50 values for Santicizer 8 Plasticizer were determined as 240, 130 and 80 mg/L respectively and are based on nominal concentrations. As a quality check, the rainbow trout were challenged with a reference compound, Antimycin A. The estimated 96 hour LC50 and 95% confidence limits (C.I.) were within the 95% confidence limits reported in the literature, indicating that the fish were in good condition .

Description of key information

Fish acute toxicity was examined in a EPA guideline (EPA-660/3 -75 -009) study under quality control. In this study 10 rainbow trout (salmo gairdneri) per concentration, were exposed to nominal concentrations of 56, 100, 180, 320, 560 and 1000 mg/L under static conditions for 96 hours. The 24 hour, 48 hour and 96 hour LC50 values for Santicizer 8 Plasticizer were determined as 240, 130 and 80 mg/L respectively and are based on nominal concentrations. As a quality check, the rainbow trout were challenged with a reference compound, Antimycin A. The estimated 96 hour LC50 and 95% confidence limits (C.I.) were within the 95% confidence limits reported in the literature, indicating that the fish were in good condition .

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
80 mg/L

Additional information

There are 2 similar studies performed in the same laboratory in the same period. Species were rainbow trout and bluegill sunfish of which rainbow trout proved most sensitive. This value has therefore been selected as key value.