7,7-dimethyl-3-oxa-6-azaoctan-1-ol

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

10 April 2001 to 12 April 2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of relevant results.

Qualifier:

according to guideline

Guideline:

other: ISO TC147/SC5/WG2

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of signature: 15/05/2001

Analytical monitoring:

no

Details on sampling:

Not applicable.

Vehicle:

no

Details on test solutions:

-Preparation of test solutions:
The behaviour of the test substance was examined in seawater according to the method described in the Harmonised Offshore Chemical Notification Format (OSPAR, 1995).

A nominal 1,000mg/l solution was prepared in dilution water, shaken vigorously and allowed to stand for 1 hour.

A clear solution was obtained. Test concentrations were prepared by addition of specified quantities of the sample to dilution water to obtain the required copncentrations.

A preliminary study had identified the 48 hour LC50 as being approximately 100mg/l and therefore definitive test concentrations were prepared as 0 (control), 25, 50, 100, 200 and 400mg/l.

Test organisms (species):

other aquatic crustacea: Acartia tonsa

Details on test organisms:

TEST ORGANISM
-Test organism:
Acartia tonsa

-Source:
Guernsey Sea Farms

-Date Acartia delivered to laboratory:
10 April 2001

-Age of copepods at start of test:
11-13 days

-Life stage:
Adult

Holding conditions of culture
-Prior to despatch:
Temperature: 20±1 deg C
Salinity: 33±2 ppt

Feeding regime:
Mixed algal diet recommended by Guernsey Sea Farms.

The Acartia tonsa for the test were cultured under semi-static conditions and were fed every working day on a mixed algal diet supplied by Guernsey Sea Farms.

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not applicable.

Hardness:

Not reported.

Test temperature:

at 0 hours:
19.5 deg C

at 48 hours:
20 deg C

pH:

at 0 hours:
control group: 8.0
400mg/l group: 9.5

at 48 hours:
control group: 8.1
400mg/l group: 8.5

Dissolved oxygen:

at 0 hours: (%ASV)
control group: 100
400mg/l group: 101

at 48 hours: (%ASV)
control group: 97
400mg/l group: 97

Salinity:

at 0 hours: (%0)
control group: 35
400mg/l group: 35

at 48 hours: (%0)
control group: 35
400mg/l group: 35

Nominal and measured concentrations:

Nominal concentrations:
0, 25, 50, 100, 200 and 400mg/l

Measured concentrations:
0, 25.1, 50.6, 101.6, 200.8, 400.0mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable):
closed

- Material, size, headspace, fill volume:
50ml crystallising dishes, fill volume 25ml.

- Aeration:
None.

- No. of organisms per vessel:
5

- No. of vessels per concentration (replicates):
4

- No. of vessels per control (replicates):
4

TEST MEDIUM / WATER PARAMETERS
The stocks of animals were maintained, and the test performed, in natural seawater supplied by Guernsey Sea Farms. The dilution water was stored in the dark at approximately 15 deg C within the testing facility. Immediately before use, the duilution water was aerated until the dissolved oxygen concentrations reached saturation (>80% ASV) and the pH stabilised at 8.0 ± 0.3.

The seawater was filtered to 5µm prior to despatch.

The dilution water used for sulturing the copepods used in this study and for the test was supplied by Guernsey Sea Farms and was delivered on 10 April 2001.

OTHER TEST CONDITIONS
- Photoperiod:16 hours light/8 hours dark.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The number of dead copepods at each test concentration were measured at 24 and 48 hours after initiation of the test.

TEST CONCENTRATIONS
-Definitive study:
0, 25, 50, 100 and 400mg/l

- Range finding study
- Test concentrations:
Not reported.

- Results used to determine the conditions for the definitive study:
Yes.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

239 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Remarks on result:

other: 95% CL 209-273 mg/l

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

201 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Details on results:

One (5%) of the twenty control copepods, died during the study. This indicates a satisfactory level of "health" of organisms maintained under test conditions.

The test substance nominal concentrations were prepared as "water-accommodated fractions" according to the OSPAR (1995) guidelines. There was no chemical analytical confirmation of the actual dissolved concentrations. The dissolved concentrations were likely to be as given in this report as the test substance was soluble in water. The water quality measurements of the test solutions are given in the test conditions section and were within acceptable limits.

Results with reference substance (positive control):

The test organism sensitivity was also monitored during this study using the recommended reference substance 3,5 DCP. Of the 20 copepods, 11 died when exposed to a 1mg/l concentration of 3,5 DCP for 48 hours at 20 deg C and therefore the 55% mortality represents an acceptable level of sensitivity (i.e. the 48 hour mortality was within the range of 20 to 80%).

Reported statistics and error estimates:

The 48 hour LC50 value of the test substance to Acartia tonsa was estimated by ToxCalc analysis to be 239mg/l as determined by the Spearman-Karber method. The highest no observed effect concentration (NOEC) after 48 hours was 201mg/l, as determined by Fisher's Exact test. The lowest test concentration which killed all twenty copepods within 48 hours was 400mg/l.

Table 2. Cumulative percent mortalities.

Actual concentration (mg/l)	Number dead		% mortality
	24 hours	48 hours	24 hours	48 hours
0	0	1	0	5
25.1	0	0	0	0
50.6	0	1	0	5
101.6	0	0	0	0
200.8	3	5	15	25
400.0	20	20	100	100
3,5-Dichlorophenol*	3	11	15	55

*A 1mg/l concentration of 3,5 -dichlorophenol was used as the reference substance to check the sensitivity of the test organisms.

Graphs 1 and 2 illustrate the estimation of the LC₅₀ values.

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of the test the substance cannot be classified according to 54/687/EEC as the 48 hour LC50 does not meet the criteria for classification.

Executive summary:

Method:

Test species: Acartia tonsa

Test type: Acute toxicity: 48 hour LC50

Test conditions: Static test conditions according to SOP E207 based upon ISOTC147/SC5/WG2 protocol by Thompson (1990): Water Quality: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea).

Test period: From 10 to 12 April 2001

Test performed at: Chemex International plc, Bar Hill Business Park, 37 Saxon Way, Bar Hill, Cambridge, CB3 8EL, England.

Results:

24 hour LC50: 256mg/l (229 -285mg/l)*

48 hour LC50: 239mg/l (209 -273mg/l)*

48 hour NOEC: 201mg/l

48 hour 100% mortality: 400mg/l

%mortality of Acartia tonsa exposed to 1mg/l 3,5 -dichlorophenol after 48 hours = 55%.

Conclusion:

The 48 hour LC50 (239mg/l) does not meet the criteria stated in 54/687/EEC and is therefore not classified.

*Denotes 95% confidence limits.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 June 1996 to 05 june 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of relevant results.

Qualifier:

according to guideline

Guideline:

other: PARCOM Ring Test Protocol (Acute Toxicity to the Marine Copepod Acartia tonsa).

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: UK proposal to ISO TC 147/SC 4/WG2: (Determination of Acute Lethal Toxicity to Marine Copepods).

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of signature: 07/06/96

Analytical monitoring:

no

Details on sampling:

Not applicable.

Vehicle:

no

Details on test solutions:

The test substance is soluble in water, and the sample preparation method for water soluble test substances was followed.

Water soluble test substanes
1) Dissolve the test substance in the filtered and aerated seawater. The concentration of this stock solution should correspond with the highest test dosage.
2) Mix the stock solution in a 1L erlenmeyer flask on a magnetic stirrer (~100 rpm) for 2-4 hours. Cover the tope with parafilm to minimize volatization. Wrap the erlenmeyer flask in aluminium foil to avoid light exposure.
3) if anything is flotated or precipitated, the Study Director should be contacted.
4) Prepare the required test dosages with dilutions of the stock solution using graduated pipettes and cylindric/volumetric measuring flasks. Use culture medium for dilutions.

Based on the results from the preliminary test, the following test concentratsion (nominal whole product) were used for the definitive test: 2.5; 25; 250; and 1000 mg/l.

Test organisms (species):

other aquatic crustacea: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Source:
University of Copenhagen, Laboratory of marine biology, Helsingør, Denmark

- Age at study initiation (mean and range, SD):
25 days

- Feeding during test
No feeding during test.


ACCLIMATION
Not stated.


QUARANTINE (wild caught)
Not applicable.

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not applicable.

Hardness:

Not reported.

Test temperature:

19.3 - 21.4°C

pH:

Not reported.

Dissolved oxygen:

7.3 mg/l
Test medium was aerated prior to disolution of test substance.

Salinity:

Not reported.

Nominal and measured concentrations:

Nominal concentrations:
2.5, 25, 250 and 1000 mg/l.

Details on test conditions:

Lethality testing with the calamoid copepod Acartia tonsa was performed twice. Initially, a preliminary (range finding) test was performed in duplicate for each test dosage followed by a definitive test with 4 parallels for each dosage.

Percentage lethality after 24 and 48 hours was determined, and the LC50, LC90, LC10 and NOEC values were calculated. LC50 is defined as the test dosage which kills 50% of the animals. NOEC is defined as the highest test dosage of the test substance which gives no observed effect on the lethality.

The test procedure is according to Aquateam's Standard Operating Procedure S.O.P. Lab II - 1.1 (Aquateam, 1994) which is based upon a UK proposal to ISO TC 147/SC5/WG2, and supplementary information from the PARCOM Ring Test Workshops in 1990 (Bjørnestad et al., 1990; PARCOM, 1990) and 1991 (PARCOM, 1992). The test system is in accordance with the SFT and PARCOM requirements (OSPAR, 1995).

References:
Aquateam (1994) Standard Operating Procedure. Test Methods. Acartia tonsa (Marine) 48 h Lethality Test. S.O.P. Lab II 1.1, Date 1 August 1994.

Bjørnestad, E; Nyholm, N and Robson, M (1990) Notes from the PARCOM Ring Test Workshop in Copenhagen, November 21-22, 1990.

PARCOM (1990) Acute Toxicity to the Marine Copepod Acartia tonsa. Paris Commission - Ring Test Protocol, WQI.

PARCOM (1992) Follow-up workshop to the PARCOM Ring Test. Summary Report. Wageningen, 6-9 January 1992.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

60 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks on result:

other: 95% CL 45-47 mg/l

Details on results:

Lethality-dosage curves for the test substance are presented in figures 1 and 2 (attached). The raw data for the definitive test are presented in Appendix 2 (attached).

The LC50-value was found to be 450 mg/l after 24 hours. The 95% confidence limit was 225-675 mg/l. The LC10-value was estimated to 40 mg/l, the LC90-value was estimated to 1000 mg/l and the NOEC-value was estimated to 2.5 mg/l after 24 hours.

The LC50-values was found to be 60 mg/l after 48 hours. The 95% confidence limit was 45-75 mg/l. The LC10-value was estimated to 2.5 mg/l, the LC-90-value was estimated to 170 mg/l after 48 hours. The test concentrations were too high to calculate NOEC value after 48 hours.

Results with reference substance (positive control):

One test concentration (1.0 mg/l) of the reference substance 3,5-dichlorophenol was analysed 03.06.96 - 0.5.06.96. The lethality of 1.0 mg/l 3,5-dichlorophenol was 40% after 48 h.

Reported statistics and error estimates:

Method for data handling:
The mean values of LC50, LC10, LC90, NOEC and the confidence limits are calculated using the computer program LOTUS 1-2-3 for Windows 4.0. The scattered half-logarithmic dosage-response diagram is printed out and the curve itself drawn by hand.

Validity criteria fulfilled:

yes

Remarks:

In this test, the control parameters yielded the following values: Dissolved oxygen concentration: 7.3 mg/l, % lethality of the control: 0%, Lethality of 1.0 mg/l 3,5-dichlorophenol: 40%, Temperature variations during the test: 19.3-21.4 deg C.

Conclusions:

The toxicity of the test substance, measured as LC50 after 48 hours, was found to be 60 mg/l. The 95% confidence limit was: 45-75 mg/l. According to the EEC classification system (EEC, 1993), a test substance is classified as very toxic if LC50 < 1 mg/l, as toxic if LC50 + 1-10 mg/l and as harmful if LC50 >10-100 mg/l. Based on this, the test substance is classified as harmful.

Executive summary:

Acartia tonsa testing has been performed on the test substance based on a UK proposal to ISO TC 147/SC 4WG2: "Determination of Acute Lethal Toxicity to Marine Copepods" and PARCOM Ring Test Protocol: "Acute Toxicity to the Marine Copepod Acartia tonsa".

The LC₅₀ -value for the test substance after 48 h was found to be 60 mg/l. The 95% confidence limit was 45 -75 mg/l. The LC₁₀ -value was estimated to 2.5 mg/l and the LC₉₀ -value was estimated to 170mg/l.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 December 1982 to 18 December 1982.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect rthe quality of relevant results.

Qualifier:

according to guideline

Guideline:

other: E 729-80 (ASTM Standard Practice for Conducting Acute Toxicity Test with Fishes, Macroinvertebrates and Amphibians)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA-660/3-75-009 (Methods for Acute Toxicity Test with Fish, Macroinvertebrates and Amphibians)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of most recent inspection: 18/01/83 Date of signature: 28/01/83

Analytical monitoring:

no

Details on sampling:

Not applicable.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
Standard operating procedure L.2.2 was utilised. SOP details not recorded.

- Controls:
Blank (test media) control

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name:
Water flea

- Strain:
Not stated.

- Source:
In-house cultures

- Age at study initiation (mean and range, SD):
<36 hours old

- Method of breeding:
Daphnids were produced from parents which had been accilimated approximately 11 days.

Reproductive populations had been acclimated for a minimum of 30 days.

- Feeding during test
No feeding during test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

None.

Hardness:

CaCO3 160 mg/l

Test temperature:

21±1 deg C

pH:

7.5-8.2

Dissolved oxygen:

At 0 hours:
0 mg/l group:8.4-8.5
39 mg/l group: 8.5-8.6
65 mg/l group: 8.5-8.6
108 mg/l group:8.5
180 mg/l group:8.4-8.5
300 mg/l group: 8.3

At 48 hours:
0 mg/l group: 9.0-9.1
39 mg/l group: 8.9
65 mg/l group: 8.9-9.0
108 mg/l group: 8.8-9.0
180 mg/l group:8.8-8.9
300 mg/l group: 8.9-9.0

Salinity:

Not applicable.

Nominal and measured concentrations:

Nominal concentrations only, no verification performed.
Nominal: 0, 39, 65, 108, 180 and 300mg/l

Details on test conditions:

TEST SYSTEM
No information reported.

TEST MEDIUM / WATER PARAMETERS
No information reported.

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light / 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
In the range finding and Definitive tests immobilisation observations were made and recorded daily. Standard operating proceedure L.2.7 was follwed.

TEST CONCENTRATIONS
- Test concentrations:
0, 39, 65, 108, 180 and 300

- Range finding study
- Test concentrations:
Not reported.

- Results used to determine the conditions for the definitive study:
Yes.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

113.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL 78.3-164.6

Details on results:

- Behavioural abnormalities:
See Table 1 (attached) for daily observation data.

-Immobilisation:
1 Daphnid did not survive in thefirst control group at the 48 hour observation.
See Table 1 (attached) for daily observation data.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

Analysis were performed based on probit transformations of Litchfield, J.T., and F Wilcoxon, 1949. These analyses were performed by a computer program writeen by M.J. Nicolich version 06-Dec-82. This program has been archived in the East Laboratory

The definitive test was perfromed two times due to poor dose response data in the first test. The raw data from the first test are available in the archived study data book.

Validity criteria fulfilled:

not specified

Conclusions:

Based upon the 48 hour EC50 value (113.6mg/l) the substance cannot be classified for the environment according to 67/548/EEC.

Executive summary:

Introduction.

This study was conducted in order to evaluate the acute toxicity of MRD-82 -178 to the daphnid (Daphnia magna).

The study was conducted in general accordance to appropriate ASTM Standards (E 729 -80) and EPA Guidelines (EPA-660/3 -75 -009).

This study was performed in the Environmenal Toxicology Laboratory, Bio/dynamics Incorporated, P.O. Box 43, East Millstone, New Jersey 08873.

Method.

Once quarantine period was over, and animals were deemed healthy, testing began. The initial test was a range finding series. Initial test concentrations were assigned by the study director. After the 24 hour range finding study was completed, the 48 hour definitive study was initiated.

The definitive test was perfromed two times due to poor dose response data in the first test. The raw data from the first test are available in the archived study data book.

Results.

Time Period	EC50	95% Confidence Limits
24 hours	122.9 mg/l	85.9 to 175.7
48 hours	113.6 mg/l	78.3 to 164.6

Conclusion.

Based upon the 48 hour EC50 value the substance cannot be classified for the environment according to 67/548/EEC.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not reported in the data over 12 years old provided by ECHA.

Reliability:

other: Reliability of study not reported in the data over 12 years old provided by ECHA.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

not specified

GLP compliance:

yes

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Test type:

not specified

Water media type:

not specified

Limit test:

no

Total exposure duration:

48 h

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

not specified

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

560 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

not specified

1000 mg/l was the highest concentration tested. It was considered unnecessary to test at higher concentrations.

Validity criteria fulfilled:

yes

Conclusions:

EC50 determined to be >1000 mg/l. Substance not classified.

Description of key information

Four acute toxicity studies on aquatic invertebrates are available (two freshwater study in Daphnia magna and two marine studies in Acartia tonsa).
The freshwater Daphnia magna studies gave a 48hr EC50 values of 113.6 mg/l and >1000 mg/l.
The marine studies in Aractia tonsa,gave 48hr EC50 values of 60 mg/l and 239 mg/l. 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

113.6 mg/L

Marine water invertebrates

Marine water invertebrates

Effect concentration:

60 mg/L