N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

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:

key study

Study period:

From February 25, 2002 to September 26, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

see 'Principles of method if other than guideline'

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

see 'Principles of method if other than guideline'

Principles of method if other than guideline:

The only deviation was that analytical quantification of the applied test concentrations was not possible due to the absence of a suitable analytical method. Therefore only the stock solution used to prepare the test concentrations were analyzed. However, based on the high water solubility and stability of the test substance it was considered reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.

GLP compliance:

yes

Specific details on test material used for the study:

Triamine content 85.3%, Total amine content 100%

Analytical monitoring:

yes

Details on sampling:

Quantification of the actual exposure concentrations in the test solutions was not possible due to the absence of suitable analytical method. Instead the concentrations in the stock solutions were quantified.

Vehicle:

yes

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION:
- Dispersion: A stock solution was prepared by dissolving approximately 1 g of test substance in 120 mL deionized water at 50 °C under stirring. After cooling pH was adjusted to 8.5 and deionized water was added up to make up a volume of 1000 mL (at Day 2 a fresh solution was prepared to change the medium). The test solutions were prepared by addition of the required amounts of stock solution to the test medium (Dutch Standard Water). - Vehicle, solvent: deionized water
- Concentration of vehicle/ solvent: 1 g/L


STABILITY OF THE TEST CHEMICAL SOLUTIONS:
- Reference substance: Dilution water
- Source: Dutch Standard Water; Containing per liter of dionized water 100 mg Na HCO3, 20 mg KHCO3, 200 mg CaCl2.2H2O and 180 mg MgSO4.7H2O
- Aeration: test solutions were aerated with water-saturated air purified by an active coal filter and cotton filter.
- Hardness: 12°dH
- pH: 8.2
- Oxygen content: 8.2-8.7 mg/L
- Conductance: < 5uS/cm
- Holding water: Dutch Standard Water (DSW)

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISMS:
- Strain: Brachydanio rerio (Zebra fish)
- Supplier: Retailer (fish were accepted according to the quality control criteria of the guidelines and SOP)
- Wild caught: no
- Age/size/loading: Juvenile/<= 3 cm/1g/L (wet weight)
- Feeding: Commercially available dry or deep-frozen food
- Pretreatment: 1-3 times a day
- Feeding during test: No; feeding stopped 24 h before study

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

no

Hardness:

Hardness: 12°dH

Test temperature:

19.4-21.4°C

pH:

8.2

Dissolved oxygen:

Oxygen content: 8.2-8.7 mg/L

Salinity:

Dutch Standard Water was used which is a freshwater

Nominal and measured concentrations:

Concentrations: 0.1, 0.18, 0.32, 0.58 and 1.05 mg/L

Details on test conditions:

TEST SYSTEM:
- Test type: Semi static
- Concentrations: 0.1, 0.18, 0.32, 0.58 and 1.05 mg/L
- Renewal of test solution: day 2
- Exposure vessel type: 5 L aquaria containing 3 L of test solution
- Number of replicates, fish per replicate: one aquarium per dose with 7 fish
- Test temperature: 19.4-21.4°C
- Dissolved oxygen: 8.2-8.7 mg/L
- pH: 7.6-8.2
- Adjustment of pH: The pH of the stock solution (1.0031 g in 120 mL and 1.0004 g in 120 mL) of the chemical prepared in deionized water of approximately 50°C after cooling down the pH was adjusted to 8.5. Deionized water was added up to a volume of 1000 mL.
- Intensity of irradiation: Ambient light provided by fluorescent tubes
- Photoperiod: 16 h photoperiod daily

DURATION OF THE TEST: 96 h

TEST PARAMETER: The fish were considered dead when a lack of opercular movement was observed and touching of the caudal peduncle produced no reaction. Dead fish were removed from the test vessels directly after being observed, which was checked at least twice each day. In addition to death, sub-lethal effects such as erratic swimming, loss of reflex, increased excitability, lethargy, changes in physiology, discoloration, pigmentation, excessive mucous production, hyperventilation, opaque eyes, curved spine or hemorrhaging were recorded. Fish that were convulsing or showing other severe forms of distress not considered transient in nature and likely to become more severe before the exposure is terminated, were sacrificed for humane reasons. These fish were treated as having died in the test.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.431 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: (0.346 - 0.538)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

1.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: observed after 29 h

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- The LC50 (96 h) was calculated to be 0.431 mg/L with 95 % confidence limits of 0.346 and 0.538 m/L.
- The highest concentration causing no mortality (no observed effect concentration, NOEC) after 96 h amounted to 0.18 mg/L whereas 0.32 m/L was the lowest tested concentration (LOEC,96 h) at which lethality was observed as compared to the control.
- The LC100 was 1.05 mg/L, which was observed after 29 h.

Reported statistics and error estimates:

The LC50 was calculated using the computer program Toxcalc v 5.0 applying the trimmed Spearman-Karber method

Sublethal observations / clinical signs:

RESULTS: EXPOSED

- Nominal/measured concentrations: 0, 0.1, 0.18, 0.32, 0.58,1.05 mg/L (nominal)

Table 1

Survival of fish

 

Nominal	Hours of test duration
Concentration	0	2	5	21	24	29	45	48	53	54	69	72	77	96
(mg/l)	Number of living fish
Control	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.1	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.18	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.32	7	7	7	7	7	7	7	7	7	7	7	7	6	6
0.58	7	7	7^a	7^a	7^a	7^a	7^a	6^a	4^a*	4^a	3	2	1	1
1.05	7	7^a	7^b	1^b	1^b	0

^a= reduced activity

^b= very reduced activity

* = For ethical reasons two fish were euthanised at the concentration of 0.58 mL/L after 53 h.

 

Table 2

Data evaluation

 

Hours of test	LC50-values	95%-confidence intervals
duration	(mg/l)	(mg/l)
24	0.82	0.7369	0.9124
48	0.7169	0.6126	0.8389
72	0.5105	0.4168	0.6253
96	0.4314	0.3460	0.5378

 

- The LC50 (96 h) was calculated to be 0.431 mg/L with 95 % confidence limits of 0.346 and 0.538 m/L. - The highest concentration causing no mortality (no observed effect concentration, NOEC) after 96 h amounted to 0.18 mg/L whereas 0.32 mg/L was the lowest tested concentration (LOEC,96 h) at which lethality was observed as compared to the control. - The LC100 was 1.05 m/L, which was observed after 29 hours.

RESULTS: CONTROL

- Number/percentage of animals showing adverse effects: NoneBased on the following observations, the validity criteria was considered as fulfilled:  - Mortality of control animals was <10 %;- The concentration of dissolved  oxygen in all test vessels was > 60 % saturation and - The concentration of  test substance >= 80 % of initial concentration during test. Although quantification of the applied concentrations was not possible  due to the absence of a suitable analytical method it was possible to analyze the two stock solutions used to prepare the test concentrations. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. Considering the  high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.  A dose-response related increase in mortality was observed.

Table 3

Oxygen measurements

 

Nominal	Days of measurements
concentration	0	1	2 old medium	2 fresh medium	3	4
(mg/l)	O2(mg/l)
Control	8.6	8.8	8.3	8.3	8.5	8.8
0.1	8.7	8.8	8.7	8.2	8.6	8.1
0.18	8.7	8.8	8.7	8.2	8.6	8.8
0.32	8.7	8.8	8.7	8.3	8.7	8.7
0.58	8.7	8.9	8.5	8.3	8.6	8.3
1.05	8.7	8.9

 

 

Table 4

pH measurements

 

Nominal	Days of measurements
concentration	0	1	2 old medium	2 fresh medium	3	4
(mg/l)	pH
Control	8.0	7.8	8.0	8.2	8.0	7.8
0.1	8.1	7.8	8.0	8.2	8.0	7.6
0.18	8.2	7.8	8.0	8.2	8.0	7.9
0.32	8.2	7.8	8.1	8.2	8.0	7.9
0.58	8.2	7.8	8.0	8.2	8.0	7.8
1.05	8.2	7.8

 

 

Table 5

Results of the chemical analyses

 

	Measured Concentration in 10 times diluted stock solution (mg/l)	Average measured concentration (mg/l)	Standard deviation	Concentration of undiluted stock solution (mg/l)
First analyses stock 25022002	111.40	110.81	0.83	1108.10
Second analyses 25022002	110.22
First analyses stock 27022002	101.54	100.01	2.17	1000.10
Second analyses 27022002	98.47

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance were calculated to be 0.43 and 0.18 mg/L, respectively. The test substance was found to be toxic to Zebra fish after 96 h at concentration levels of 0.32 mg/L and higher.

Executive summary:

A study was conducted to determine the acute toxicity of the test substance to Zebra fish (Danio rerio) according to the OECD Guideline 203 and EU Method C.1, in compliance with GLP. In this study, single replicate of seven fish per concentration were exposed to 0.1, 0.18, 0.32, 0.58 and 1.05 mg/L nominal test concentrations under semi-static conditions for 96 h. Mortality was observed at 24, 48, 72 and 96 h intervals. Although quantification of the applied concentrations was not possible due to the absence of a suitable analytical method, it was possible to analyze the two stock solutions used to prepare the test concentrations. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility and stability of the test substance, it was considered reasonable to assume that the actual concentrations would be identical to the nominal concentrations during the entire test. All the validity criteria were considered to be fulfilled. Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance were calculated to be 0.43 and 0.18 mg/L, respectively. The test substance was found to be toxic to Zebra fish after 96 h at concentration levels of 0.32 mg/L and higher (Company data, 2002).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Lepomis macrochirus

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

22±2°C

pH:

8.1 to 8.3

Dissolved oxygen:

> 7.5 mg/L (86% of saturation)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.45 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the mean measured 96 h LC50 and NOEC of the test substance were determined to be 0.45 and 0.3 mg/L, respectively.

Executive summary:

A study was conducted to determine the toxicity of the test substance to bluegill sunfish (Lepomis macrochirus) according to EPA OPPTS Guideline 850.1075, in compliance with GLP. Toxicity, expressed a mortality and clinical signs, followed a positive dose-response relationship. Dose levels were verified analytically. All validity criteria for the study were fulfilled. Under the study conditions, the mean measured 96 h LC50 and NOEC of the test substance were determined to be 0.45 and 0.3 mg/L, respectively (Drottar, 2000).

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No analytical dose verification

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1400 (Fish Acute Toxicity Test)

Deviations:

yes

Remarks:

no analytical verification of test concentrations.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.75 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.58 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

no

Remarks:

no analytical veryfication

Conclusions:

Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance were determined to be 0.75 and 0.58 mg/L, respectively

Executive summary:

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to EPA OTS Guideline 797.1400, in compliance with GLP. Toxicity, expressed a mortality and clinical signs, followed a positive dose-response relationship. No analytical dose verification was conducted. Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance were determined to be 0.75 and 0.58 mg/L, respectively (Holmes, 1992).

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1460 (Fish Acute Toxicity Test mitigated by humic acid)

Deviations:

yes

Remarks:

No analytical verification of test concentrations.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Details on test conditions:

Humic acid content: 10 mg humic acid/litre.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

no

Remarks:

no analytical verification

Conclusions:

Under the study conditions, the 96 h LC50 and NOEC of the test substance mitigated by dissolved organic carbon were determined to be 3.0 and 1.4 mg/L (nominal), respectively. This study demonstrates the mitigating effects of organic matter on the test substance.

Executive summary:

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to EPA OTS 797.1460 (fish acute toxicity test mitigated by humic acid) under static conditions, in compliance with GLP. The applied humic acid content in this experimental setup was 10 mg/L. No analytical dose verification was conducted. The toxicity of the test substance to fish followed a positive dose-response relationship for mortality and clinical signs of toxicity. Under the study conditions, the 96 h LC50 and NOEC of the test substance mitigated by dissolved organic carbon were determined to be 3.0 and 1.4 mg/L (nominal), respectively (Holmes, 1992).

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1460 (Fish Acute Toxicity Test mitigated by humic acid)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Details on test conditions:

Humic acid content: 20 mg humic acid/litre

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

3.1 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

no

Remarks:

no analytical veryfication

Conclusions:

Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance mitigated by dissolved organic carbon were determined to be 3.0 mg/L and 1.4 mg/L (nominal), respectively. This study shows the positive relationship between the concentration of organic matter and mitigating effect on the test substance.

Executive summary:

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to EPA OTS 797.1460 (fish acute toxicity test mitigated by humic acid) under static conditions, in compliance with GLP. The applied humic acid content in this experimental setup was 20 mg/L. No analytical dose verification was conducted. The toxicity of the test substance to fish followed a positive dose-response relationship for mortality and clinical signs of toxicity. Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance mitigated by dissolved organic carbon were determined to be 3.0 mg/L and 1.4 mg/L (nominal), respectively (Holmes, 1992).

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Not all validity criteria met

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Remarks:

derivation and quantification by UV absorbance

Vehicle:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.68 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.23 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

4.44 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Toxicity of the test substance to fish followed a positive dose-response relationship for mortality and clinical signs of toxicity.
- Results from analytical part:
Recovery of the test substance from the test medium was variable, with concentrations recovered at the lower test concentrations falling below 80% after 48 or 96 hours. At other time periods the recoveries were above 100% of nominal. No indication of the possible cause of the loss of test material is stated. However, the analytical methodology (derivation and quantification by UV absorbance) could account for the deviation as at the level of quantification it is not as specific as chromatographic analysis. Further, the method of analysis is not stability indicating.It is considered that although this validation criteria has not been fulfilled the performance of the study and the result are reliable.

Validity criteria fulfilled:

no

Remarks:

It is considered that although this validation criteria has not been fulfilled the performance of the study and the result are reliable.

Conclusions:

Under the study conditions, the measured 96 h LC50 and NOEC were determined to be 0.68 and 0.23 mg/L, respectively.

Executive summary:

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203, in compliance with GLP. Animals were exposed to the test substance for 96 h under static conditions. Toxicity, expressed a mortality and clinical signs, followed a positive dose-response relationship. Analytical dose verification was conducted. Recovery of the substance from the test medium was variable, with levels at the lower test concentrations falling below 80% after 48 or 96 h. At other time periods, the recoveries were above 100%. Although validity of the test method was not evaluated, performance of the study and the result were considered to be reliable. Under the study conditions, the measured 96 h LC50 and NOEC were determined to be 0.68 and 0.23 mg/L, respectively (Ritter, 1989).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.43 mg/L

Additional information

A study was conducted to determine the acute toxicity of the test substance to Zebra fish (Danio rerio) according to the OECD Guideline 203 and EU Method C.1, in compliance with GLP. In this study, single replicate of seven fish per concentration were exposed to 0.1, 0.18, 0.32, 0.58 and 1.05 mg/L nominal test concentrations under semi-static conditions for 96 h. Mortality was observed at 24, 48, 72 and 96 h intervals. Although quantification of the applied concentrations was not possible due to the absence of a suitable analytical method, it was possible to analyze the two stock solutions used to prepare the test concentrations. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility and stability of the test substance, it was considered reasonable to assume that the actual concentrations would be identical to the nominal concentrations during the entire test. All the validity criteria were considered to be fulfilled. Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance were calculated to be 0.43 and 0.18 mg/L, respectively. The test substance was found to be toxic to Zebra fish after 96 h at concentration levels of 0.32 mg/L and higher (Company data, 2002).

A study was conducted to determine the toxicity of the test substance to bluegill sunfish (Lepomis macrochirus) according to EPA OPPTS Guideline 850.1075, in compliance with GLP. Toxicity, expressed a mortality and clinical signs, followed a positive dose-response relationship. Dose levels were verified analytically. All validity criteria for the study were fulfilled. Under the study conditions, the mean measured 96 h LC50 and NOEC of the test substance were determined to be 0.45 and 0.3 mg/L, respectively (Drottar, 2000).

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203, in compliance with GLP. Animals were exposed to the test substance for 96 h under static conditions. Toxicity, expressed a mortality and clinical signs, followed a positive dose-response relationship. Analytical dose verification was conducted. Recovery of the substance from the test medium was variable, with levels at the lower test concentrations falling below 80% after 48 or 96 h. At other time periods, the recoveries were above 100%. Although validity of the test method was not evaluated, performance of the study and the result were considered to be reliable. Under the study conditions, the measured 96 h LC50 and NOEC were determined to be 0.68 and 0.23 mg/L, respectively (Ritter, 1989).

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to EPA OTS Guideline 797.1400, in compliance with GLP. Toxicity, expressed a mortality and clinical signs, followed a positive dose-response relationship. No analytical dose verification was conducted. Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance were determined to be 0.75 and 0.58 mg/L, respectively (Holmes, 1992a).

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to EPA OTS 797.1460 (fish acute toxicity test mitigated by humic acid) under static conditions, in compliance with GLP. The applied humic acid content in this experimental setup was 10 mg/L. No analytical dose verification was conducted. The toxicity of the test substance to fish followed a positive dose-response relationship for mortality and clinical signs of toxicity. Under the study conditions, the 96 h LC50 and NOEC of the test substance mitigated by dissolved organic carbon were determined to be 3.0 and 1.4 mg/L (nominal), respectively (Holmes, 1992b).

A study was conducted to determine the toxicity of the test substance to rainbow trout (Oncorhynchus mykiss) according to EPA OTS 797.1460 (fish acute toxicity test mitigated by humic acid) under static conditions, in compliance with GLP. The applied humic acid content in this experimental setup was 20 mg/L. No analytical dose verification was conducted. The toxicity of the test substance to fish followed a positive dose-response relationship for mortality and clinical signs of toxicity. Under the study conditions, the nominal 96 h LC50 and NOEC of the test substance mitigated by dissolved organic carbon were determined to be 3.0 mg/L and 1.4 mg/L (nominal), respectively (Holmes, 1992c).