Fatty acids, tall-oil, compds. with triethanolamine

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 April 2001 - 14 June 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Read across justification in attached in section 13

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Remarks:

During the study a deviation in pH by >1 unit was observed, in contrast to the protocol. This deviation was not considered to have affected the study outcome

Qualifier:

according to guideline

Guideline:

other: OECD Paris (2000), Guidence Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures. OECD Series on Testing and Assessment, No. 23. ENV/JM/MONO 6.

Version / remarks:

OECD Paris (2000), Guidence Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures. OECD Series on Testing and Assessment, No. 23. ENV/JM/MONO 6.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source: Pine Chemicals Association, Inc.
- Appearence: Viscous brown liquid
- Batch No.of test material: 5-10-00-17
- Expiration date of the batch:15 May 2006
- Purity test date: No Certificate of Analysis or details regarding test item purity were supplied. Study conducted using the test substance as received.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was poorly soluble and therefore was mixed to create a Water Accommodated Fraction (WAF) with loading rates of 1000, 750, 500, 250, 125 and 0 mg/L. WAF was prepared in 2 litre capacity glass vessels with sidearm, with 2 litres of ISO freshwater algal medium, these were then placed on a magnetic stirrer. The appropriate weight of test item was added to this stirring medium and the vessels sealed to avoind contamination and loss of valatile fractions. The stirring speed adjusted to give a stiring vortex of 5-10% of the water column. After ca. 48 hours of stirring, the medium was allowed to settle for ca.1 hour and the aqueous phase removed. The first ca.100 ml was discarded before ca. 800 ml was collected as test medium. Control medium, without the addition of test item, was stirred and extracted in an identical manner to treated media, using the same volume of extract. A volume of ca. 50 ml was added to the appropriate test vessels.

Analytical monitoring:

no

Details on sampling:

- Concentrations: 1000, 750, 500, 250, 125 and 0 mg.l-1
- Sampling method: Algal cell concentrations were observed in the test solutions of each flask at 24, 48 and 72 hours after initiation (definitive test). Aliquots were removed by pipette and cell concentrations determined using a Compound Light Microscope and Improved Neubauer Counting Chambers.
- Sample storage conditions before analysis: Not specified, although test flasks were maintained in a cooled oribital incubator to maintain a temperature of 22-26 ºC (maximum range 2ºC per vessel). Throughout the study duration, algal cells were rotated orbitally at 100 rpm with a light intensity range of between 6400 -9600 lux.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test item was poorly soluble and therefore was mixed to create a Water Accommodated Fraction (WAF) with loading rates of 1000, 750, 500, 250, 125 and 0 mg.l-1. WAF was prepared in 2 litre capacity glass vessels with sidearm, with 2 litres of ISO freshwater algal medium, these were then placed on a magnetic stirrer. The appropriate weight of test item was added to this stirring medium and the vessels sealed to avoind contamination and loss of valatile fractions. The stirring speed adjusted to give a stiring vortex of 5-10% of the water column. After ca. 48 hours of stirring, the medium was allowed to settle for ca.1 hour and the aqueous phase removed. The first ca.100 ml was discarded before ca. 800 ml was collected as test medium. Control medium, without the addition of test item, was stirred and extracted in an identical manner to treated media, using the same volume of extract. A volume of c. 50 ml was added to the appropriate test vessels.

- Controls: Water Accommodated Fraction prepared in exact way as test solutions, with the test item not being present.

Test organisms (species):

Selenastrum sp.

Details on test organisms:

TEST ORGANISM
- Common name: Selenastrum capricornutum
- Strain: 278/4
- Source (laboratory, culture collection): Starter cultures (Strain 278/4) obtained (2001) from Culture Centre of Algae and Protozoa (CCAP), Ambleside, Cumbria, UK.
- Age of inoculum (at test initiation): Strain was received as a starter culture (15th February 2001). Experimental commencement was the 2nd May 2001.
- Method of cultivation: Transfers of the alga were made regularly into fresh algal growth medium to provide suitable anexic subcultures, in the exponential phase of growth for test inoculants.

ACCLIMATION
- Culturing media and conditions (same as test or not): Same

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test temperature:

24-26ºC

pH:

6.78 - 9.31

Nominal and measured concentrations:

Nominal concentrations of 1000, 750, 500, 250, 125 and 0 mg.l-1

Details on test conditions:

TEST SYSTEM
- Test vessel: Water Accomodated Fractions were prepared in 2 litre capacity glass vessels with a sidearm, with 2 litres of ISO freshwater algal medium. these were then placed on a magnetic stirrer with a stir bar. the requisite quantity of test material was added to the stirring medium and the vessels sealed to prevent contamination and avoid loss of volatile fractions. After a 48 hour stirring period they were left to stand for 1 hour and the aqueous phase removed via the vessel sidearm. 800 mL of test medium was extracted for use as test medium. Control medium, without the addition of test item was extracted under the same methodology, using the same volume of extract. A volume of 50 mL was added to each test vessel.

- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: 2 litre Glass vessels of ISO freshwater algal medium
- Initial cells density: 10^4 cells.ml-1
- No. of organisms per vessel: 10^4 cells.ml-1
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Alkalinity: pH 6.78 - 9.31
- Culture medium different from test medium: no
- Intervals of water quality measurement: At the start and end of the definitive test (0h and 72h)

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Light intensity and quality: 8830 lux (definitive test)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Cell concentrations determined using a compound light microscope and improved neubauer counting chamber.

TEST CONCENTRATIONS
- Range finding study: yes
- Test concentrations: 1000, 100, 10, 1 and 0 mg.l-1.
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

no

Duration:

72 h

Dose descriptor:

EL50

Remarks:

Based on Area Under Curve (AUC)

Effect conc.:

ca. 854.9 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: AUC

Basis for effect:

cell number

Key result

Duration:

72 h

Dose descriptor:

EL50

Remarks:

Average Specific Growth rate

Effect conc.:

> 1 000 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: Average Specific Growth Rate

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

NOELR

Effect conc.:

ca. 500 mg/L

Nominal / measured:

meas. (initial)

Basis for effect:

other:

Remarks:

AUC

Duration:

72 h

Dose descriptor:

NOELR

Effect conc.:

ca. 750 mg/L

Nominal / measured:

meas. (initial)

Basis for effect:

growth rate

Details on results:

- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): none

Reported statistics and error estimates:

AUC (0-72hr) EL50 (ppm) estimate 854.90 mg.l-1. 95% Cl (821.19, 891.30 mg.l-1)

Mean Cell Concentration (x10⁵cells.ml^-1) at 0, 24, 48 and 72h During the Definitive Test

		Initial Loading Rate of Fatty acids, tall oil (mg.l^-1)
Time (h)	Replicate/Mean	1000	750	500	250	125	0
0	Mean	0.1	0.1	0.1	0.1	0.1	0.1
24	I	0.15	0.40	0.13	0.35	0.75	0.28
24	II	0.15	0.08	0.43	0.65	0.50	0.75
24	III	0.13	0.28	0.30	0.38	0.58	0.50
24	IV	-	-	-	-	-	0.30
24	V	-	-	-	-	-	0.60
24	VI	-	-	-	-	-	0.35
24	Mean	0.14	0.25	0.29	0.46	0.61	0.46
48	I	0.75	2.93	4.98	4.40	4.58	3.43
48	II	0.78	2.50	4.00	4.35	4.35	4.88
48	III	0.48	2.28	3.28	4.50	5.50	5.43
48	IV	-	-	-	-	-	4.45
48	V	-	-	-	-	-	5.60
48	VI	-	-	-	-	-	4.08
48	Mean	0.67	2.57	4.09	4.41	4.81	4.65
72	I	4.43	12.33	20.45	17.08	15.00	17.80
72	II	4.98	14.10	19.40	20.35	13.85	12.95
72	III	5.53	9.28	11.80	17.48	16.60	14.50
72	IV	-	-	-	-	-	12.75
72	V	-	-	-	-	-	15.70
72	VI	-	-	-	-	-	14.30
72	Mean	4.98	11.90	17.22	18.30	15.15	14.67

- = Three flasks tested

Cell Growth Group Mean Values Determined from Daily Cell Counts at 24, 48 and 72h During the Definitive Test

		Initial Loading Rate of Fatty acids, tall oil (mg.l^-1)
Growth Function	Time Interval (h)	1000	750	500	250	125	0
Area Under Growth Curve (AUC) (x 10^5cells.h.ml^-1)	0 -24	0.36	0.81	0.94	1.49	1.79	1.47
Area Under Growth Curve (AUC) (x 10^5cells.h.ml^-1)	0 -48	0.94	1.62	1.85	1.89	1.93	1.91
Area Under Growth Curve (AUC) (x 10^5cells.h.ml^-1)	0 -72	1.30	1.59	1.71	1.74	1.67	1.66
Average Specific Growth Rate (µave.day^-1)	0 -24	0.02	0.08	0.09	0.18	0.26	0.18
Average Specific Growth Rate (µave.day^-1)	0 -48	0.16	0.70	1.09	1.26	1.43	1.32
Average Specific Growth Rate (µave.day^-1)	0 -72	1.02	2.84	4.24	4.59	4.25	4.06

Values of pH Measured For Test Solutions at 0 and 72h During the Definitive Test

		Initial Loading Rate of Fatty acids, tall oil (mg.l^-1)
Time/Replicate		1000	750	500	250	125	0
0h	I	6.78	6.94	7.06	7.19	7.27	7.87
72h	I	8.03	8.36	8.42	8.38	8.71	8.65
72h	II	8.02	8.18	8.41	8.39	8.75	8.67
72h	III	8.02	8.09	8.17	8.31	8.40	9.31
72h	IV	-	-	-	-	-	9.00
72h	V	-	-	-	-	-	8.69
72h	VI	-	-	-	-	-	8.71

- = Three flasks tested

Validity criteria fulfilled:

not specified

Remarks:

Although not stated in the report, validity criteria appear to have been sufficiently met.

Conclusions:

The 72h EL50 for AUC was 854.90 mg.l-1 with a corresponding NOELr of 500 mg.l-1, under test conditions. The 72h EL50 based on average specific growth rate was > 1000 mg.l-1, the highest loading rate tested. However, the corresponding NOELr was 500 mg.l-1 at 0 -48h, and 750 mg/L at 0 -72h indicating some inhibition (<50%) compared to the control.

Executive summary:

The study was designed to determine the median effective loading (EL50) of Fatty acids, Tall oil, CAS No. 61790 -12 -3, by exposing alga (Selenastrum capriconutum) to solutions of prepared Water Accommodated Fractions (WAF) with test item loading rates of 1000, 750, 500, 250, 125 and 0 mg.l-1. It was performed based on procedures in OECD (1984) Guideline 201, and following procedures in OECD (2000) Guidance Document No. 23 on Testing Difficult Substances, according to GLP.

During the definitive test, environmental conditions were within the following ranges, pH: 6.78 -9.31 and temperature: 24 -26°C. The light intensities were 9450 lux (range finding test) and 8830 lux (definitive test).

The 72h EL50 for the area under the curve (AUC) was 854.90 mg.l-1 with a corresponding NOELr of 500 mg.l-1, under test conditions. The 72h EL50 based on Average Specific Growth Rate was > 1000 mg.l-1, the highest loading rate tested, with a corresponding NOELr of 750 mg.l-1.

This guideline, GLP-compliant study has been evaluated as being reliable without restriction.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Remarks:

Accordning to In a German Industrial Standard DIN 38412 part 9. USEPA ECOTOX database evaluation: Reliability: 2A: “Remarks: Reliable with restrictions; Acceptable, well-documented publication report which meets basic scientific principles.”. Study also considered valid for use in OECD SIDS review for triethanolamine.

Justification for type of information:

Read across justification in attached in section 13

Qualifier:

according to guideline

Guideline:

other:

Version / remarks:

DIN 38412, Part 9 (German National Guideline - Acute static growth inhibition)

Principles of method if other than guideline:

DIN 38412, Part 9 (German National Guideline - Acute static growth inhibition)

GLP compliance:

not specified

Specific details on test material used for the study:

CAS Number: 102-71-6
Identity: Triethanolamine
Purity: Not stated

Analytical monitoring:

no

Vehicle:

not specified

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

Species identified as Scenedesmus subspicatus

Test type:

static

Water media type:

not specified

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

72 h measurement also conducted

pH:

Toxicity tests have been carried out at both constant pH and allowing the pH to increase with increasing triethanolamine concentration.

Reference substance (positive control):

not specified

Key result

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

7.9 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: solutions not neutralised

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

216 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: solutions not neutralised

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

26 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: solutions neutralised

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

512 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: solutions neutralised

Duration:

72 h

Dose descriptor:

EC90

Effect conc.:

5 900 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: solutions not neutralised

Duration:

72 h

Dose descriptor:

EC90

Effect conc.:

10 100 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: solutions neutralised

Duration:

96 h

Dose descriptor:

EC10

Effect conc.:

7.1 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other:

Remarks:

solutions not neutralised

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

169 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other:

Remarks:

solutions not neutralised

Duration:

96 h

Dose descriptor:

EC90

Effect conc.:

4 030 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other:

Remarks:

solutions not neutralised

Duration:

96 h

Dose descriptor:

EC10

Effect conc.:

13.2 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other:

Remarks:

solutions neutralised

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

910 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other:

Remarks:

solutions neutralised

Duration:

96 h

Dose descriptor:

EC90

Effect conc.:

62 500 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other:

Remarks:

solutions neutralised

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour acute toxicity of the test substance to Scenedesmus subspicatus has been adequately characterized. The EC50 (growth rate, 72 h) was 216 mg/L for the non-neutralised test and 512 mg/L for the neutralised test. The EC10 (growth rate, 72 h) was 7.9 mg/L for the non-neutralised and 26 mg/L for the neutralised test.

Executive summary:

In a study according to German Industrial Standard DIN 38412 part 9 the 96- hour acute toxicity of the test substance to Scenedesmus subspicatus has been adequately characterized. Full details of the study are not available but the study was considered valid according to an entry to the USEPA ECOTOX database evaluation (Reliability: 2A: “Remarks: Reliable with restrictions; Acceptable, well-documented publication report which meets basic scientific principles.”). The study was also considered valid for use in OECD SIDS review for triethanolamine.

The EC50 (growth rate, 72 h) was 216 mg/L for the non-neutralised test and 512 mg/L for the neutralised test. The EC10 (growth rate, 72 h) was 7.9 mg/L for the non-neutralised and 26 mg/L for the neutralised test.

Description of key information

The acute toxicity of the target substance, Tall oil, compound with triethanolamine, to green algae, was assessed by read-across from its components, tall oil & triethanolamine. Based on read-across data, the target substance is not expected to be hazardous to Algae, with the lowest measured 72-h EC50 (nominal) of 216 mg/L. The read-across justification document covering this and other endpoints is attached to Section 13.

Key value for chemical safety assessment

EC50 for freshwater algae:

216 mg/L

EC10 or NOEC for freshwater algae:

7.9 mg/L