Soaps, stocks, vegetable-oil, acidulated

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:

weight of evidence

Study period:

March 2002 to April 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ISO 14669, ISO 10253 and GESAMP protocol for preparation of poorly soluble complex mixtures

Deviations:

no

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

Samples were taken for carbon analysis at 0, 24 and 48 h.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accommodated fraction (WAF) methodology
- Preparation: WAFs were prepared by adding test substance to the test medium (sterilised natural seawater) to give loading rates of 10, 100 and 1,000 mg/L. In order to mix the test substance, which was solid at room temperature, the sample was microwaved on full power for approximately 2 min until it became liquid. The contents of the WAF vessels were then stirred using a magnetic stirrer, at 300 rpm, for 24 h, in a controlled environment room set at 20 ± 2°C and then left to settle for a minimum of 2 h (range 2 – 4 h), to allow the undissolved material to separate out. The aqueous phases - the WAFs – were then drawn off via the tap for use in the tests. Control media was sterilised seawater.

Test organisms (species):

other aquatic crustacea: Acartia tonsa

Details on test organisms:

TEST ORGANISMS
- Age: 12-14 d old.
- Source: Obtained from Guernsey Sea Farms, Vale, Guernsey

ENVIRONMENTAL CONDITIONS
- Temperature: 20-22°C
- Light: 16 h light/8 h dark cycle
- Diet: Fed ad libitum with mixed marine algae

Test type:

static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20.5-20.7°C

pH:

8.2 - 8.1

Dissolved oxygen:

6.1 - 6.2 mg/L

Salinity:

32-33%

Nominal and measured concentrations:

Nominal: 0 and 1,000 mg/L

Details on test conditions:

-All media and glassware were autoclaved at 121°C for 15 min before use.
-A preliminary test was made to decide stirring speed. A speed of 300 rpm was selected.
-2 vessels where used during the test, containing 2 L of media and known volume of test item.
-The tests were conducted in 150 mL glass crystal dishes containing 100 mL of either control or test solution. A. tonsa were added using modified glass Pasteur pipettes. A. tonsa were considered to be immobilised if they failed to respond to touching with the end of a sealed glass pipette. At the end, a few drops of formalin were added. Immobilisation, carbon concentration and growth were estimated.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

% immobilised after exposure:
24 h: 3%
48 h: 3%

Results with reference substance (positive control):

% immobilised after exposure:
24 h: 22%
48 h: 53%

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 48 h EL50 of the constituent was determined to be >1,000 mg/L.

Executive summary:

A study was conducted to evaluate the acute toxicity of the constituent glycerides C16 -18, C18 -unsatd (palm oil) to Acartia tonsa under static conditions. The test method was based on guidelines issued by ISO 14669, ISO 10253 and GESAMP. Acartia tonsa were exposed to water accomodated fractions prepared from the constituent at concentrations of 0 and 1,000 mg/L and immobility was determined at 0, 24 and 48 h. 3,5 -dichlorophenol was used as a reference substance. Under the test conditions, the 48 h EL50 of the constituent was determined to be >1000 mg/L (Worden and Sherren, 2002b).

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

March 2002 to April 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ISO 14669, ISO 10253 and GESAMP guidelines

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

Samples were taken for carbon analysis at 0, 24 and 48 h.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficultly soluble test substances)
- Method: Water accommodated fraction (WAF) methodology
- Preparation: WAFs were prepared by adding test substance to the test medium (sterilised natural seawater) to give loading rates of 10, 100 and 1,000 mg/L. The contents of the WAF vessels were then stirred using a magnetic stirrer, at 300 rpm, for 24 h, in a controlled environment room set at 20 ± 2°C and then left to settle for a minimum of 2 h (range 2 – 4 h), to allow the undissolved material to separate out. The aqueous phases - the WAFs – were then drawn off via the tap for use in the tests. Control media was sterilised seawater.

Test organisms (species):

other aquatic crustacea: Acartia tonsa

Details on test organisms:

TEST ORGANISMS
- Age: 12-14 d old.
- Source: Obtained from Guernsey Sea Farms, Vale, Guernsey

ENVIRONMENTAL CONDITIONS
- Temperature: 20-22°C
- Light: 16 h light/8 h dark cycle
- Diet: Fed ad libitum with mixed marine algae

Test type:

static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20-22°C

pH:

8.5-8.2

Dissolved oxygen:

6.1-6.2 mg/L

Salinity:

32-33%

Nominal and measured concentrations:

Nominal: 0 and 1,000 mg/L

Details on test conditions:

- All media and glassware were autoclaved at 121°C for 15 min before use.
- A preliminary test was made to decide stirring speed. A speed of 300 rpm was selected.
- Two vessels where used during the test, containing 2 L of media and known volume of test item. The test was conducted on 150 mL glass crystallising dishes containing 100 mL of either control or test solution.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

% immobilised after exposure:
24 h: 0%
48 h: 3%

Results with reference substance (positive control):

% immobilised after exposure:
24 h: 22%
48 h: 53%

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 48 h EL50 of the constituent was determined to be >1,000 mg/L.

Executive summary:

A study was conducted to evaluate the acute toxicity of the constituent glycerides C16 -18, C18 -unsatd. (soybean oil) to Acartia tonsa under static conditions. The test method was based on the guidelines ISO 14669, ISO 10253 and GESAMP. Acartia tonsa were exposed to water accomodated fractions prepared from the constituent at concentrations of 0 and 1,000 mg/L and immobility was determined at 0, 24 and 48 h. 3,5 -dichlorophenol was used as a reference substance. Under the test conditions, the 48 h EL50 of the constituent was determined to be >1,000 mg/L (Worden and Sherren, 2002a).

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

January 12 to 17, 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

other: ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)

Deviations:

no

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accommodated fraction (WAF) methodology
- Preparation: WAFs were prepared by adding test substance to the test medium (sterilised natural seawater) to give loading rates of 10, 100 and 1000 mg/L. In order to mix the test substance, which was solid at room temperature, the sample was microwaved on full power for approximately 2 min until it became liquid. The contents of the WAF vessels were then stirred using a magnetic stirrer, at 300 rpm, for 24 h, in a controlled environment room set at 20 ± 2°C and then left to settle for a minimum of 2 h (range 2 – 4 h), to allow the undissolved material to separate out. The aqueous phases (the WAFs) were then drawn off via the tap for use in the tests. Control media was sterilised seawater.

Test organisms (species):

other: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Source: Guernsey Sea Farms, Vale, Guernsey
- Age at study initiation (mean and range, SD): 9-12 d
- Feeding
- Food type: Mixed marine algae supplied with the organisms
- Amount, frequency: Ad libitum


ACCLIMATION
- Acclimation period: 1 d
- Acclimation conditions (same as test or not): Temperature: 18-20°C

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20±2°C

pH:

8.1-8.6

Dissolved oxygen:

6.4-8.3 mg/L

Salinity:

33-34%

Nominal and measured concentrations:

Nominal concentration: 0, 10, 100 & 1,000 mg/L
- 1,000 mg/L was equivalent to 4 mg total carbon/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: Glass crystallising dishes, 150 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): Not renewed
- No. of organisms per vessel: 32 (control), 35 (control, filtered), 31-33 (test material) and 33 (positive control)
- No. of vessels per concentration (replicates): Three
- No. of vessels per control (replicates): Three
- No. of vessels per vehicle control (replicates): Three


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Natural seawater, collected off the beach at Kinmel Bay to the west of Rhyl, Flintshire, N. Wales
- Total organic carbon: 28 & 29 mg/L for filtered and unfiltered controls; 25, 27 & 31 mg/L for test substance at 10, 100 & 1,000 mg/L, respectively


OTHER TEST CONDITIONS
- Photoperiod: 16 h light/8 h dark cycle
- Temperature: 20±2°C

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol; 1 mg/L

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

>= 100 - < 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

mobility

Details on results:

Observation of immobilised organisms under a low power, binocular microscope did not reveal any adverse signs and is therefore concluded that the effects seen were due to toxicity.

Results with reference substance (positive control):

% immobilisation: 21 (after 24 h); 76 (after 48 h)

Preliminary study: The results from the study indicated that the test material was toxic to Acartia tonsa at a loading rate of 1000 mg/L.

Table 1: Immobilisation ofA. tonsaexposed to WAF of coconut oil and 1 mg/L solution of reference compound 3,5-DCP

Test material	Loading rate (mg/L)	Number of A. tonsa exposed	Cumulative Number (N) and Percentage (P) of A. tonsa immobilised after exposure period of:
			24 h		48 h
			N	P	N	P
Control		32	0	0	0	0
Control (filtered)		35	0	0	2	6
Coconut oil	10	31	0	0	3	10
	100	32	0	0	0	0
	1000	33	2	6	17	52
3,5-DCP	1	33	7	21	25	76

Validity criteria fulfilled:

yes

Conclusions:

Based on the results, the 48 h EL50 of the constituent was determined to be between 100 and 1,000 mg/L.

Executive summary:

A study was conducted to evaluate the acute toxicity of the constituent "Glycerides C8 -18, C18 -unsatd (coconut oil) to Acartia tonsa under static conditions. The method followed the guideline ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea). Acartia tonsa were exposed to water accommodated fractions prepared from the constituent at concentrations ranging from 10-1,000 mg/L for 48 h and immobility was determined at 24 and 48 h. Based on the results, the 48 h EL50 of the constituent was determined to be between 100 and 1,000 mg/L (Worden and Sherren, 2004b).

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)

Deviations:

no

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accommodated fraction (WAF) methodology
- Preparation: WAFs were prepared by adding test substance to the test medium (sterilised natural seawater) to give loading rates of 10, 100 and 1000 mg/L. In order to mix the test substance, which was solid at room temperature, the sample was microwaved on full power for approximately 2 min until it became liquid. The contents of the WAF vessels were then stirred using a magnetic stirrer, at 300 rpm, for 24 h, in a controlled environment room set at 20 ± 2°C and then left to settle for a minimum of 2 h (range 2 – 4 h), to allow the undissolved material to separate out. The aqueous phases - the WAFs – were then drawn off via the tap for use in the tests. Control media was sterilised seawater.

Test organisms (species):

other: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Source: Guernsey Sea Farms, Vale, Guernsey
- Age at study initiation (mean and range, SD): 13-15 d
- Food type: Mixed marine algae supplied with the organisms
- Amount, frequency: Ad libitum

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 2°C

pH:

7.7-8.3 (at time 0), 8.1-8.2 (after 48 h)

Dissolved oxygen:

8.6-8.9 mg/L (at time 0), 7.6-8.9 (after 48 h)

8.6-8.9 mg/L (at time 0), 7.6-8.9 (after 48 h)

Salinity:

32-33%

Nominal and measured concentrations:

Nominal concentration: 10, 100 and 1,000 mg/L

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: Glass crystallising dishes, 150 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): Not renewed
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): Three
- No. of vessels per control (replicates): Three
- No. of vessels per vehicle control (replicates): Three


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Natural seawater, collected off the beach at Kinmel Bay to the west of Rhyl, Flintshire, N. Wales
- Total organic carbon (mean of three replicates at time 0): 28.1, 29.1 and 33.2 mg/L for 10, 100 and 1000 mg/L loading rate, respectively


OTHER TEST CONDITIONS
- Photoperiod: 16 h light/8 h dark cycle
- Temperature: 20 ± 2°C

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol; 1 mg/L

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The WAF prepared from palm kernel oil was found to be non-toxic to A. tonsa, with 48 h EL50 values exceeding 1,000 mg/L, the highest loading rate tested.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Other: The percentage of A. tonsa that were immobilised when exposed to 1 mg/L 3,5-DCP was 58% after 48 h.

Table 1: Immobilisation of A. tonsa exposed to WAF of palm kernel oil and 1 mg/L solution of reference compound 3,5-DCP

Test material	Loading rate (mg/L)	Number of A. tonsa exposed	Cumulative Number (N) and Percentage (P) of A. tonsa immobilised after exposure period of:
			24 h		48 h
			N	P	N	P
Control		35	1	3	4	11
Palm kernel oil	10	35	0	0	6	17
	100	34	1	3	3	9
	1000	32	2	6	5	16
3,5-DCP	1	33	4	12	19	58

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 48 h EL50 of the test substance was determined to be >1,000 mg/L.

Executive summary:

A study was conducted to evaluate the acute toxicity of the constituent C8 -18, C18 -unsatd. (palm kernel oil) to Acartia tonsa under static conditions. The method followed the guideline ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea). Acartia tonsa were exposed to water accomodated fractions prepared from the constituent at concentrations ranging from 10 to 1,000 mg/L for 48 h and immobility was determined at 24 and 48 h. Under the test conditions, the 48 h EL50 of the constituent was determined to be >1,000 mg/L (Worden and Sherren, 2004d).

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)

Deviations:

no

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficultly soluble test substances)
- Method: Water accommodated fraction (WAF) methodology
- Preparation: WAFs were prepared by adding test substance to the test medium (sterilised natural seawater) to give loading rates of 10, 100 and 1000 mg/L. In order to mix the test substance, which was solid at room temperature, the sample was microwaved on full power for approximately 2 min until it became liquid. The contents of the WAF vessels were then stirred using a magnetic stirrer, at 300 rpm, for 24 h, in a controlled environment room set at 20 ± 2°C and then left to settle for a minimum of 2 h (range 2 – 4 h), to allow the undissolved material to separate out. The aqueous phases - the WAFs – were then drawn off via the tap for use in the tests. Control media was sterilised seawater.

Test organisms (species):

other: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Source: Guernsey Sea Farms, Vale, Guernsey
- Age at study initiation (mean and range, SD): 13-15 d
- Food type: Mixed marine algae supplied with the organisms
- Amount, frequency: Ad libitum

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 2°C

pH:

7.7-8.3 (at time 0), 8.1-8.2 (after 48 h)

Dissolved oxygen:

8.6-8.9 mg/L (at time 0), 7.6-8.9 (after 48 h)

Salinity:

32-33%

Nominal and measured concentrations:

Nominal concentration: 10, 100 and 1,000 mg/L

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: Glass crystallising dishes, 150 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): Not renewed
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): Three
- No. of vessels per control (replicates): Three
- No. of vessels per vehicle control (replicates): Three


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Natural seawater, collected off the beach at Kinmel Bay to the west of Rhyl, Flintshire, N. Wales
- Total organic carbon (mean of three replicates at time 0): 28.4, 29.7 and 33.9 mg/L for 10, 100 and 1000 mg/L loading rate, respectively


OTHER TEST CONDITIONS
- Photoperiod: 16 h light/8 h dark cycle
- Temperature: 20 ± 2°C

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol; 1 mg/L

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The WAF prepared from palm kernel olein was found to be non-toxic to A. tonsa, with 48 h EL50 values exceeding 1,000 mg/L, the highest loading rate tested.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Other: The percentage of A. tonsa that were immobilised when exposed to 1 mg/L 3,5-DCP was 58% after 48 h.

Table 1: Immobilisation of A. tonsa exposed to WAF of palm kernel olein and 1 mg/L solution of reference compound 3,5-DCP

Test material	Loading rate (mg/L)	Number of A. tonsa exposed	Cumulative Number (N) and Percentage (P) of A. tonsa immobilised after exposure period of:
			24 h		48 h
			N	P	N	P
Control		35	1	3	4	11
Palm kernel olein	10	31	1	3	7	23
	100	34	1	3	2	6
	1000	35	0	0	2	6
3,5-DCP	1	33	4	12	19	58

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 48 h EL50 of the constituent was determined to be >1,000 mg/L.

Executive summary:

A study was conducted to evaluate the acute toxicity of the constituent C8 -18, C18 -unsatd. (palm kernel olein) to Acartia tonsa under static conditions. The method followed the guideline ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea).

Acartia tonsa were exposed to water accomodated fractions prepared from the constituent at concentrations ranging from 10 to 1,000 mg/L for 48 h and immobility was determined at 24 and 48 h. Under the test conditions, the 48 h EL50 of the constituent was determined to be >1,000 mg/L (Worden and Sherren, 2004c).

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)

Deviations:

no

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accommodated fraction (WAF) methodology
- Preparation: WAFs were prepared by adding test material to the test medium (sterilised natural seawater) to give loading rates of 10, 100 and 1,000 mg/L. In order to mix the test substance (solid at room temperature), the sample was microwaved on full power for approximately 2 min until it became liquid. The contents of the WAF vessels were then stirred using a magnetic stirrer, at 300 rpm, for 24 h, in a controlled environment room set at 20 ± 2°C and then left to settle for a minimum of 2 h (range 2 – 4 h), to allow the undissolved material to separate out. The aqueous phases - the WAFs – were then drawn off via the tap for use in the tests. Control media was sterilised seawater.

Test organisms (species):

other: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Source: Guernsey Sea Farms, Vale, Guernsey
- Age at study initiation (mean and range, SD): 13-15 d
- Food type: Mixed marine algae supplied with the organisms
- Amount, frequency: Ad libitum

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 2°C

pH:

7.7-8.3 (at time 0), 8.1-8.2 (after 48 h)

Dissolved oxygen:

8.6-8.9 mg/L (at time 0), 7.6-8.9 (after 48 h)

Salinity:

32-33%

Nominal and measured concentrations:

Nominal concentrations: 0, 10, 100 & 1,000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: Glass crystallising dishes, 150 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): Not renewed
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): Three
- No. of vessels per control (replicates): Three
- No. of vessels per vehicle control (replicates): Three


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Natural seawater, collected off the beach at Kinmel Bay to the west of Rhyl, Flintshire, N. Wales
- Total organic carbon (mean of three replicates at time 0): 28.3, 28.6 and 29.2 mg/L for 10, 100 and 1000 mg/L loading rate, respectively


OTHER TEST CONDITIONS
- Photoperiod: 16 h light/8 h dark cycle
- Temperature: 20 ± 2°C

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol; 1 mg/L

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

- Results with reference substance valid: yes
- Other: The percentage of A. tonsa that were immobilised when exposed to 1 mg/L 3,5-DCP was 58% after 48 h.

Table 1: Immobilisation of A. tonsa exposed to WAF of palm kernel stearin and 1 mg/L solution of reference compound 3,5-DCP

Test material	Loading rate (mg/L)	Number of A. tonsa exposed	Cumulative Number (N) and Percentage (P) of A. tonsa immobilised after exposure period of:
			24 h		48 h
			N	P	N	P
Control		35	1	3	4	11
Palm kernel stearin	10	33	2	6	3	9
	100	36	1	3	5	14
	1000	39	5	13	10	26
3,5-DCP	1	33	4	12	19	58

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 48 h EL50 of the constituent was determined to be > 1,000 mg/L.

Executive summary:

A study was conducted to evaluate the acute toxicity of the constituent palm kernel stearin to Acartia tonsa under static conditions. The procedures followed ISO 14669:1999 "Water quality" Part 5: Biological methods Section 5.24: Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea). Acartia tonsa were exposed to water accommodated fractions prepared from the constituent at concentrations ranging from 10 to 1,000 mg/L for 48 h and immobility was determined at 24 and 48 h. Under the test conditions, the 48 h EL50 of the constituent was determined to be > 1,000 mg/L (Worden and Sherren, 2004a).

 

Reference 7

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

23 March 2010 to 21 July 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Refer to the section 13 for details on the read across justification. The acute fish study with the read across substance is used as a supporting study.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Series on testing and assessment No. 23: Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
- Sampling method: The concentrations of the test substance were analysed in the duplicate test media samples from both sampling times (0 and 48 h). From the control samples only one of the duplicate samples was analysed from each of both sampling times.
- Sample storage conditions before analysis: All samples were stored in a freezer (≤ - 10°C), protected from light until analysis was performed

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 19.9, 23, 10.3, 9.4 and 4.3 mg of the test substance were added directly to 1990, 5000, 4900, 9900 and 10000 mL test water, respectively, and carefully stirred for about 24 h at room temperature in the dark to dissolve as much of the test substance as possible. After cessation of mixing and a following period of settling of about 1 h and 45 min to allow phase separation, the aqueous phases, i.e. the water accommodated fractions, were drawn off carefully and used as the test media of the nominal test concentrations.
- Controls: Yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Elendt "M4" reconstituted water
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The appearance of the test substance in the test media was observed at the start of the test and after 24 and 48 h test duration in the only test concentration. No remarkable observations were made.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Clone 5
- Source: In house laboratory culture (i.e. IBACON testing laboratory)
- Age at study initiation (mean and range, SD): 6.25 to 21.5 h old
- Method of breeding: The cultivation of the parental Daphnia was performed in reconstituted water of a similar quality with regards to pH, the constituent salts and total hardness as the test water used in the test. The test organisms were not first brood progeny.
- Food type: Green algae (Desmodesmus subspicatus) freshly grown in the lab
-Frequency: Daily

ACCLIMATION
- Was not necessary, since the test was performed in the same medium as the culturing

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

20 - 22°C

pH:

7.7- 7.9

Dissolved oxygen:

8.0 - 9.0 mg/L

Nominal and measured concentrations:

Nominal: 0, 0.43, 0.95, 2.1, 4.6 and 10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- Type (delete if not applicable): Closed with lids
- Material, size, headspace, fill volume: Glass beakers of 100 mL volume containing approximately 60 mL of test medium.
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 20 Daphnia per test concentration and control, divided into 4 groups of 5 animals, each group in 50 mL test medium



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt "M4" reconstituted water
- Alkalinity: 0.9 mmol/L
- Culture medium different from test medium: No


OTHER TEST CONDITIONS
- Adjustment of pH: Yes
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 940 - 1150 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility of the Daphnia was determined by visual observation after 24 and 48 h. Those animals not able to swim within 15 sec after gentle agitation of the test beaker were considered to be immobile (even if they could still move their antennae).


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: Yes
A dose-related effect of the test material was observed in an additional range-finding test for the water accommodated fractions of nominal 10 and 1.0 mg/L. Nominal test concentrations 10, 4.6, 2.1, 0.95 and 0.43 mg/L were tested in the main experiment.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.95 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

2.1 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

5.17 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

0.95 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

LOEC

Effect conc.:

2.1 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: No
- Mortality of control: No
- After 48 h of exposure no immobilisation of the test animals was observed in the control and up to the nominal test substance concentration of 0.95 mg/L. At 2.1 mg/L, one daphnid was immobile after 48 h of exposure. 8 immobile daphnids were observed at nominal 4.6 mg/L. At the highest concentration of 10 mg/L, 18 daphnids were immobile.

Therefore, the 48 h NOEC was determined to be 0.95 mg/L. The 48 h LOEC was determined to be 2.1 mg/L. The 48 h EC50 value was calculated to be 5.17 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid: Yes
- 48 h EC50: 1.37 mg/L indicating that the sensitivity of the Daphnia was consistent with the level proposed by the OECD 202 guideline
- 48 h NOEC: 0.62 mg/L
- 48 h LOEC: 1.4 mg/L
- Other: Test was not conducted parallely with this study but prior to this study.

Analytical Results:

Determination of the Test substance	Based on the results of TOC measurements the concentration of the test substance was determined using a calibration curve.
Calibration Range	1 to 50 mg Carbon/L in pure water in the case of TC-calibration and 1 to 25 mg Carbon/L in pure water in the case of IC-calibration
Linearity of Response	Correlation of peak area of different standard solutions with their corresponding concentrations, using a linear regression
Regression Coefficient	At least 0.9999 in the case of TC at least 0.9985 in the case of IC
Typical Calibration Curve	y = 4.3873 * x + 0.5934 in the case of TC y = 4.4984 * x – 1.8233 in the case of IC
Limit of Detection	LODTC: 2.26 mg Carbon/L LODIC: 2.45 mg Carbon/L
Limit of Quantification	3 mg Carbon/L 125 % (n = 5, RSD 6 %)
Mean Recovery in the Fortified Samples	118 % (n = 14, RSD 8 %)
Mean recovery in the test samples	Since the test substance is not well water soluble, all results were below LOQ

Validity criteria fulfilled:

yes

Conclusions:

Based on the results of the read across study, 48 h EC50, LOEC and NOEC for the test substance, Soaps, stocks, vegetable oil, acidulated, are considered to be 5.17, 2.1 and 0.95 mg/L WAF (nominal), respectively.

Executive summary:

A study was conducted to determine the acute toxicity of the read across substance Oils, vegetable, deodorizer distillates to Daphnia magna in a static 48 h immobilisation test according to OECD Guideline 202 and EU method C.2. Twenty Daphnia magna per dose were exposed to water accommodated fraction (WAF) of the nominal test concentrations of 0, 0.43, 0.95, 2.1, 4.6 and 10 mg/L for 48 h under static conditions. Immobility of the Daphnia was determined by visual observation after 24 and 48 h. Analytical verification of the test concentrations at the start and the end of the test via the TOC-method revealed results below LOQ. Therefore, all concentrations were represented as the nominal concentration of the read across substance. After 48 h of exposure, no immobilisation of the test animals was observed in the control and up to 0.95 mg/L. At 2.1 mg/L, one daphnid was immobile after 48 h of exposure. 8 immobile daphnids were observed at 4.6 mg/L and at the highest concentration of 10 mg/L, 18 daphnids were immobile. Therefore, under the test conditions the 48 h NOEC and LOEC were determined to be 0.95 and 2.1 mg/L respectively. The 48 h EC50 was determined to be 5.17 mg/L (Kley and Deierling, 2010c). Based on the results of the read across study, 48 h EC50, LOEC and NOEC for the test substance, Soaps, stocks, vegetable oil, acidulated, are considered to be 5.17, 2.1 and 0.95 mg/L WAF (nominal), respectively.

Reference 8

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

23 March 2010 to 26 March 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

KL2 due to RA

Justification for type of information:

Refer to the section 13 for details on the read across justification. The acute fish study with the read across substance is used as a supporting study.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Series on testing and assessment No. 23: Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
- Sampling method: The concentrations of the test substance were analysed in the duplicate test media samples from both sampling times (0 and 48 h). From the control samples only one of the duplicate samples was analysed from each of both sampling times.
- Sample storage conditions before analysis: All samples were stored in a freezer (≤ - 10°C), protected from light until analysis was performed

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficultly soluble test substances)
200.8 mg of test item were added directly to 200 mL of test water and carefully stirred for about 24 h at room temperature in the dark to dissolve as much of the test substance as possible. After cessation of stirring and a following period of settling to allow phase separation, the aqueous phase, i.e. the water accomodated fraction, was drawn off carefully and used as the test medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Clone 5
- Source: In house laboratory culture (i.e. IBACON testing laboratory)
- Age at study initiation (mean and range, SD): 6.25 to 21.5 h old
- Method of breeding: The cultivation of the parental Daphnia was performed in reconstituted water of a similar quality with regards to pH, the constituent salts and total hardness as the test water used in the test. The test organisms were not first brood progeny
- Food type: Green algae (Desmodesmus subspicatus) freshly grown in the lab
-Frequency: Daily

ACCLIMATION
- Was not necessary, since the test was performed in the same medium as the culturing

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

21°C

pH:

7.7- 7.8

Dissolved oxygen:

8.1 - 8.8 mg/L

Nominal and measured concentrations:

Nominal: 0 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- Type (delete if not applicable): Closed with lids
- Material, size, headspace, fill volume: Glass beakers of 100 mL volume containing approximately 60 mL of test medium.
- No. of organisms per vessel: 20 Daphnia per control and test concentration, divided into 4 groups of 5 animals, each group in 50 mL test medium



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt "M4" reconstituted water
- Alkalinity: 0.9 mmol/L
- Culture medium different from test medium: No


OTHER TEST CONDITIONS
- Adjustment of pH: Yes
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 940 - 1150 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility of the Daphnia was determined by visual observation after 24 and 48 h. Those animals not able to swim within 15 sec after gentle agitation of the test beaker were considered to be immobile (even if they could still move their antennae).


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: Yes
Since a dose-related effect of the test material was observed in an additional range-finding test for the water accommodated fractions of nominal 10 and 1.0 mg/L. Nominal test concentrations 10, 4.6, 2.1, 0.95 and 0.43 mg/L were tested in the main experiment.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: No
- Mortality of control: No
- Other adverse effects control: None

The concentration of test substance recorded by total organic carbon analysis was below the limit of quantification in the control and test vessels. The LOQ was equivalent to 3 mg/L. Test concentrations were therefore based on nominal values.

Validity criteria fulfilled:

yes

Conclusions:

Based on the results of the read across study, 48 h NOEC and EC50 for the test substance, Soaps, stocks, vegetable oil, acidulated, is considered to be 100 and >100 mg/L WAF (nominal), respectively.

Executive summary:

A study was conducted to determine the acute toxicity of the read-across substance Soybean oil, deodorizer distillates to Daphnia magna in a static 48 h immobilisation test (OECD Guideline 202 and EU method C.2). Twenty Daphnia magna were exposed to WAFs of the nominal test concentrations of 0 and 100 mg/L for 48 h. Immobility of the Daphnia was determined by visual observation after 24 and 48 h. Analytical verification of the test concentrations at the start and the end of the test via the TOC- method revealed results below LOQ. Therefore, all concentrations were represented as the nominal concentration of the test substance. Under the test conditions, the nominal 48 h NOEC of the read-across substance was determined to be 100 mg/L and the nominal 48 h EC 50 was >100 mg/L (Kley and Deierling, 2010d). Based on the results of the read across study, 48 h NOEC and EC50 for the test substance, Soaps, stocks, vegetable oil, acidulated, is considered to be 100 and >100 mg/L WAF (nominal), respectively.

Description of key information

Based on the studies with major constituents as well as by read-across to the similar deodorizer distillates, the lower 48 h EC50 of 5.17 mg/L and NOEC of 0.95 mg/L (nominal) has been considered further for the hazard assessment of 'soaps, stocks, vegetable oil, acidulated'.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

5.17 mg/L

Additional information

In absence of studies with ‘soaps, stocks, vegetable oil, acidulated’, its acute aquatic toxicity potential to daphnia has been evaluated on the basis of available studies with major constituents as well as by read-across to the similar deodorizer distillates.

A study was conducted to determine the acute toxicity of the read across substance Oils, vegetable, deodorizer distillates to Daphnia magna in a static 48 h immobilisation test according to OECD Guideline 202 and EU method C.2. Twenty Daphnia magna per dose were exposed to water accommodated fraction (WAF) of the nominal test concentrations of 0, 0.43, 0.95, 2.1, 4.6 and 10 mg/L for 48 h under static conditions. Immobility of the Daphnia was determined by visual observation after 24 and 48 h. Analytical verification of the test concentrations at the start and the end of the test via the TOC-method revealed results below LOQ. Therefore, all concentrations were represented as the nominal concentration of the read across substance. After 48 h of exposure, no immobilisation of the test animals was observed in the control and up to 0.95 mg/L. At 2.1 mg/L, one daphnid was immobile after 48 h of exposure. 8 immobile daphnids were observed at 4.6 mg/L and at the highest concentration of 10 mg/L, 18 daphnids were immobile. Therefore, under the test conditions the 48 h NOEC and LOEC were determined to be 0.95 and 2.1 mg/L respectively. The 48 h EC50 was determined to be 5.17 mg/L (Kley and Deierling, 2010c). Based on the results of the read across study, 48 h EC50, LOEC and NOEC for the test substance, Soaps, stocks, vegetable oil, acidulated, are considered to be 5.17, 2.1 and 0.95 mg/L WAF (nominal), respectively.

A study was conducted to determine the acute toxicity of the read-across substance Soybean oil, deodorizer distillates to Daphnia magna in a static 48 h immobilisation test (OECD Guideline 202 and EU method C.2). Twenty Daphnia magna were exposed to WAFs of the nominal test concentrations of 0 and 100 mg/L for 48 h. Immobility of the Daphnia was determined by visual observation after 24 and 48 h. Analytical verification of the test concentrations at the start and the end of the test via the TOC- method revealed results below LOQ. Therefore, all concentrations were represented as the nominal concentration of the test substance. Under the test conditions, the nominal 48 h NOEC of the read-across substance was determined to be 100 mg/L and the nominal 48 h EC 50 was >100 mg/L (Kley and Deierling, 2010d). Based on the results of the read across study, 48 h NOEC and EC50 for the test substance, Soaps, stocks, vegetable oil, acidulated, is considered to be 100 and >100 mg/L WAF (nominal), respectively.

Further, several acute toxicity studies were conducted on the saltwater copepod Acartia tonsa using water accommodated fractions of glyceride representative substances i.e., ‘glycerides, C8-18 and C18-unsatd.’ in the form of coconut oil, palm kernel stearin, palm kernel olein and palm kernel oil (Worden and Sherren, 2004 a and b). The lowest loading rate which caused immobilisation in 50% of the exposed animals (48 h EL₅₀) was above 100 mg/L, suggesting low acute toxicity. This is in line with results obtained with ‘glycerides, C16-18 and C18-unsatd’ (Worden and Sherren, 2002; Bowmeret al.,1998).

Overall based on the above studies, ‘soaps, stocks, vegetable oil, acidulated' is considered to have low acute toxicity potential in daphnia and the lower 48 h EC50 calculated at 5.17 mg/L has been considered further for hazard assessment.