Glycerol trioctanoate

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:

key study

Study period:

11 May - 13 May 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

EG guideline 84/449/EWG

Deviations:

yes

Remarks:

21 daphnids used in the test instead of 20 in two test concentrations

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The poorly soluble test substance was added to synthetic freshwater at 1 g/L and stirred for 18 hours. Afterwards the solution was filtrated. This solution was used as stock solution. The DOC content was determined to be 2.95 mg/L, equivalent to a test substance content of 4.38 mg/L. From this stock solution a series of test concentrations were prepared (see "Nominal and measured concentrations" field)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain: Straus, clone 5
- Source: own breeding
- Age at study initiation: < 24 hours
- Method of breeding: parthogenetic reproduction
- Feeding during test: none

ACCLIMATION/CULTURE
- Culture conditions: M4 medium according to Elendt, 1 L beakers
- Type and amount of food: Scenedesmus subspicatus
- Feeding frequency: daily
- Health during acclimation: healthy, darkbrown colour

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 1°C

pH:

7.6 - 8.1

Dissolved oxygen:

7.5 - 7.6 mg/L

Nominal and measured concentrations:

Nominal: 0.53, 0.70, 1.01, 1.40, 2.01, 2.80 and 3.94 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Fill volume: 10 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4
- Biomass loading rate: 2 mL per test organism

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: synthetic freshwater (CaCl2xH2O: 294 mg/L; MgSO4x7H2O: 123 mg/L; NaHCO3: 63 mg/L; KCl: 5.5 mg/L)

OTHER TEST CONDITIONS
- Photoperiod: dark

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 hours test duration.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: < 2

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 3.94 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: filtered test solution

Basis for effect:

mobility

Details on results:

- Immobility of control: none
- Any observations that might cause a difference between measured and nominal values: No, as stock solution was filtered and accordingly the highest tested concentration was the highest water soluble concentration of the test substance.

No effect up to the limit of the water solubility of the test substance was observed.

Table 1: Cumulative numbers of immobilised individuals during the test, absolute and % readings

Concentration [mg/L]	24 h			48 h
	mobile	immobile	immobile %	mobile	immobile	immobile %
Control	20	0	0	20	0	0
0.53	20	0	0	19	1	5
0.70*	21	0	0	21	0	0
1.01	19	1	5	19	1	5
1.40	20	0	0	20	0	0
2.01*	21	0	0	21	0	0
2.80	20	0	0	20	0	0
3.94	19	1	5	19	1	5

* 21 daphnids used in test

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8 Nov-10 November 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions (precipitation occurred in those test vessels where the substance was directly added, probably creating the observed effects by physical interference)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

. Additionally consideration of the "VCI-Konzept-Entwurfes für die (akute) aquatische Prüfung von schwerlöslichen Stoffen vom 31.1. 1994"

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Both test runs (a) directly weighed in and b) separated from undissolved test substance and)
- Sampling method: samples were taken at 0 and 48 hours

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the low solubility of the test substance, two different test solutions were prepared according to the VCI-concept for poorly soluble substances.
a) 100 mg/L directly weighed in solution: The test substance was directly weighed into the vessel and ultrasonicated for 5 min.
b) Test solution with 100 mg/L and subsequently separation of undissolved test substance: 100 mg test substance were added to 1 L M4 medium and stirred for 24h. Afterwards the undissolved test substance was separated by filtration with a fibre glass filter (pretreated with 1 molar NaOH)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: STRAUS
- Source: BGA Berlin (May 1994) afterwards own breeding
- Age at study initiation (mean and range, SD): 6 - 24h
- Feeding during test: none


ACCLIMATION
- Type and amount of food: Scenedesmus subspicatus

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

21.8°C

pH:

7.8 - 7.9

Dissolved oxygen:

94% oxygen saturation

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Measured concentrations:
Test solution a) directly weighed in without separation: 0h = 79.33 mg/L; 48h = 26.76 mg/L
Test solution b) separation of undissolved matter: 0h = 2.33 mg/L; 48 h = 0.08 mg/L The recovery of the test substance at the beginning and after 48 hours was higher than the water solubility of the test substance determined with the column elution method, what means that the separation was not complete.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL glass beaker, covered with a glass disc and filled with 50 mL
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: 5 mL per daphnia


TEST MEDIUM / WATER PARAMETERS
- Test medium: M4-medium according to guideline


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours


EFFECT PARAMETERS MEASURED: Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: Limit test - only 100 mg/L

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: solution a) test substance directly weighed in without separation of undissolved matter

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: solution b) separation of undissolved matter: No effect up to the limit of the water solubility of test substance (< 0.01 mg/L).

Details on results:

- Immobility of control: none
- Any observations that might cause a difference between measured and nominal values: Yes, for solution a) a turbidity of the test suspension was observed.
- Effect concentrations exceeding solubility of substance in test medium: yes, for solution a) without separation of undissolved test substance

No effect was observed up to the limit of the water solubility of the test substance under conditions tested. For the test solution a) without the separation of the undissolved test substance, an immobilisation of 20 to 30% (mean 25%) was observed.

Table 1: Immobilisation (%) of daphnids in solution a) (without separation of undissolved test substance)

Concentration (mg/L)	Incubation time (h)
	24	48
0/1	0	0
0/2	0	0
100/1	0*	20*
100/2	10*	30*

*Daphnia swimming on the surface, caught in oil drops. A turbidity of the test suspension was observed.

 

Table 2: Immobilisation (%) of daphnids in solution b) (with separation of undissolved test substance)

Concentration (mg/L)	Incubation time (h)
	24	48
0/1	0	0
0/2	0	0
100/1	0	0
100/2	0	0

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Aug - 24 Aug 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Remarks:

LUBW Landesanstalt für Umwelt, Messungen und Naturschutz Baden-Württemberg, Karlsruhe, Germany

Analytical monitoring:

yes

Details on sampling:

- Analysis was performed in additional test vessels without Daphnia to avoid disturbance of the measurement by the organisms itself or excrements. At the start (0 h) and at the end (48 h) of the experimental phase samples (4 mL) were taken and filtered for DOC analysis with a 0.45 µm CA membrane filter (Whatman FP 30/0.45 (im). First the filter was washed with 100 mL bi-distilled water and with about 15 mL sample. For TOC the samples were measured without filtration.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the sparingly soluble properties of the test item, the study was performed with WAFs ("water accommodated fraction") prepared with Daphnia medium similar to M4 described in OECD 202. The test item was weighted on a piece of polyethylene foil (rinsed with ethanol and H2O bidest. before) which was transferred together with the test item into a defined volume of Daphnia medium into a 1000 mL beaker. The suspension was stirred for 48 hours at 21 - 22 °C in the dark. For stirring a magnetic stirrer with a 2 cm stir bar was used. After stirring was stopped, the suspension was allowed to float and sediment for a period of 1 hour. After stopping the stirring the WAFs with higher loading rates were slightly turbid. After one hour sedimentation and flotation period no particles could be observed in suspension which could affect the organisms physically. The test solution was taken from the middle of the suspension in the beaker using a glass tube and transferred into the test vessels.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: German Federal Environmental Agency, Marienfelde, Department IV 2.4; current breeding is held at Hydrotox GmbH since February 2007
- Age at study initiation (mean and range, SD): 30 min - 21 h 30 min (gathered by separating the adults from the young animals twice within a period of 21 h 40 min)
- Method of breeding: Twice a week the animals are placed into fresh Elendt-Medium M4 (according to OECD 202). They are kept at approximately 10 animals / 200 mL. Daily, they are fed with a suspension of the algae Desmodesmus subspicatus in order to keep the C-content in the incubation beakers at 0.1 mg C per Daphnia and day. The C-content oft the algal suspension is measured photometrically taking a correlation between measured TOC and photometrical chlorophyll determination as basis. The Daphnia are held at 20 ± 2 °C with a light / dark cycle of 16 / 8 h.
- Feeding during test: none

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.9 - 21.6 °C

pH:

7.69 - 8.01

Dissolved oxygen:

7.9 - 9.2 mg O2/L

Nominal and measured concentrations:

Nominal: control, 5.8, 12.8, 28.2, 62.0, 136.4, and 300 mg/L (WAF, nominal)
Mean measured concentrations were 3.0, 15.6, 29.8 and 140.7 mg/L at nominal loading rates of 5.8, 28.2, 62 and 300 mg/L, respectively.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 50 mL glass beakers filled with 20 mL test solution
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per chemical analysis: 2

TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: no; similar to M4-medium accordig to OECD 202

OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): After 24h and 48h, the swimming capability of Daphnia magna was determined.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

218.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

102.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

136.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

300 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Details on results:

- 100% immobilization was reported at the highest nominal loading rate of 300 mg/L, whereas no immobilization was observed at any other test concentration.
- A correlation between measured loading rates and nominal loading rates for the 62 mg/L and 300 mg/L treatment groups was made. The parameters of the curve allow calculating the measured EC50 concentration.

Results with reference substance (positive control):

EC50 (<24h) = 2.88 mg/L (95% CL: 2.55 - 3.97 mg/L).
EC50 (<48h) = 1.58 mg/L (95% CL: 1.42 - 1.73 mg/L).

Results of the accompanying analysis:

The analytical results show that only 40.2 - 61.7% of the nominal TOC concentration was found in the test solutions. That means that the test item is not completely dispersible. During the 48 hours test course almost no decrease of TOC/DOC concentration was measured. Only marginal differences between 0 h and 48 h measurements occurred. In addition, only slightly higher TOC concentrations were measured compared to the DOC concentrations. TOC content of the test item was determined with 64.3%. From this the recovery rates and the measured test item concentration in the test solution were calculated.

Table: Results from the TOC/DOC analysis of the test item concentration

	A	C	D	E
Nominal loading rate [mg/L]	300	62	28.2	5.8
DOC measured 0h [mg/L]	85.2	16.6	7.9	1.7
DOC measured 48h [mg/L]	85.2	15.1	9.3	2.2
TOC measured 0h [mg/L]	89.4	16.1	10.8	2.3
TOC measured 48h [mg/L]	91.6	22.2	9.2	1.5
TOC expected [64.3% mg/L]	192.9	39.9	18.1	3.7
Recovery 0h [%]	46.3	40.4	59.6	61.7
Recovery 48h [%]	47.5	55.7	50.7	40.2
Measured loading rate 0h [mg/L]	139.0	25.0	16.8	3.6
Measured loading rate 48h [mg/L]	142.5	34.5	14.3	2.3
Measured loading rate 0-48h [mg/L]	140.7	29.8	15.6	3.0

Table: Results of the Daphnia test

	A	B	C	D	E	F	Control
Loading Rate [mg/L]	300	136.4	62.0	28.2	12.8	5.8	0
Immobile Daphnia 24h	30%	0%	0%	0%	0%	0%	0%
Immobile Daphnia 48h	100%	0%	0%	0%	0%	0%	0%

Table: Results of the Daphnia test in detail (0 h - 48 h)

Concentration test item [mg/L]	Volume test solution [mL]	[h]	Number of immobile Daphnia / number Daphnia introduced				Immobile Daphnia total	Introduced Daphnia total	Daphnia immobile [%]
			1	2	3	4
300	20	24	1/5	1/5	2/5	2/5	6	20	30
		48	5/5	5/5	5/5	5/5	20	20	100
136.4	20	24	0/5	0/5	0/5	0/5	0	20	0
		48	0/5	0/5	0/5	0/5	0	20	0
62.0	20	24	0/5	0/5	0/5	0/5	0	20	0
		48	0/5	0/5	0/5	0/5	0	20	0
28.2	20	24	0/5	0/5	0/5	0/5	0	20	0
		48	0/5	0/5	0/5	0/5	0	20	0
12.8	20	24	0/5	0/5	0/5	0/5	0	20	0
		48	0/5	0/5	0/5	0/5	0	20	0
5.8	20	24	0/5	0/5	0/5	0/5	0	20	0
		48	0/5	0/5	0/5	0/5	0	20	0
Control		24	0/5	0/5	0/5	0/5	0	20	0
		48	0/5	0/5	0/5	0/5	0	20	0

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Remarks:

Summary of available data used for the endpoint assessment of the target substance

Adequacy of study:

key study

Justification for type of information:

Please refer to the analogue justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 3.94 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: filtered test solution

Basis for effect:

mobility

Remarks on result:

other: Source: CAS 620-67-7, Sasol, 1993, Daphnia magna

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

218.2 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test material / WAF

Basis for effect:

mobility

Remarks on result:

other: Source: CAS 85536-07-8, Cremer, 2012, Daphnia magna

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

102.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Source: CAS 85536-07-8, Cremer, 2012, Daphnia magna

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

136.4 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test material / WAF

Basis for effect:

mobility

Remarks on result:

other: Source: CAS 85536-07-8, Cremer, 2012, Daphnia magna

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Source: CAS 65381-09-1, BASF, 1995, Daphnia magna; test substance directly weighed in without separation of undissolved matter

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Source: CAS 65381-09-1, BASF, 1995, Daphnia magna; separation of undissolved matter

Additional information

Since no studies investigating the short-term toxicity of Glycerol trioctanoate (CAS 538-23-8) to aquatic invertebrates are available, in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5, a read-across to the structurally related source substances Glycerol triheptanoate (CAS 620-67-7), Glycerides, C8-10 mono and di (CAS 85536-07-8), and Decanoic acid, ester with 1,2,3-propanetriol octanoate (CAS 65381-09-1) was conducted. The source substances are representative to evaluate the short-term toxicity of the target substance to aquatic invertebrates. This read-across is justified in detail within the analogue justification in IUCLID section 13.

The target substance Glycerol trioctanoate (CAS 538-23-8) is a monoconstituent characterized by the alcohol component glycerol which is fully esterified with octanoic acid to give glycerol triesters of octanoic acid.

The source substance Glycerol triheptanoate (CAS 620-67-7) is a monoconstituent characterized by the alcohol component glycerol which is esterified to give triesters of C7 fatty acids.

The source substance Glycerides, C8-10 mono and di (CAS 85536-07-8) is a UVCB substance characterized by the alcohol component glycerol and C8-10 (even numbered) fatty acids. Esterification of glycerol gives mainly mono- and diesters of C8-10 (even numbered) fatty acids.

The source substance Decanoic acid, ester with 1,2,3-propanetriol octanoate (CAS 65381-09-1) is a UVCB substance characterized by the alcohol component glycerol. Complete esterification with octanoic and decanoic fatty acids gives glycerol triesters of octanoic decanoic acid.

The study with read-across substance Glycerol triheptanoate (CAS 620-67-7) was performed in a static design and under GLP according to EU Method C.2 (Acute toxicity for Daphnia). The freshwater flea Daphnia magna was used as test organism. The daphnia were exposed to nominal concentrations of 0.53, 0.70, 1.01, 1.40, 2.01, 2.80 and 3.94 mg/L over a period of 48 hours. The highest tested concentration was the highest water soluble concentration of the test substance. No effect (immobility) was observed up to the limit of water solubility. The EC50 (48h) was determined as > 3.94 mg/L.

The study with read-across substance Glycerides, C8-10 mono and di (CAS 85536-07-8) was performed in a static design under GLP according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The freshwater flea Daphnia magna was used as test organism. The daphnia were exposed to nominal concentrations (WAF) of 5.8, 12.8, 28.2, 62.0, 136.4, and 300 mg/L over a period of 48 hours. 100% immobilization was reported at the highest nominal loading rate of 300 mg/L, whereas no immobilization was observed at any other test concentration. The EC50 (48 h) was determined as 102.6 mg/L (mean measured concentration).

The study with read-across substance Decanoic acid, ester with 1,2,3-propanetriol octanoate (CAS 65381-09-1) was performed as a limit test in a static design and under GLP according to EU Method C.2 (Acute toxicity for Daphnia). The freshwater flea Daphnia magna was used as test organism. The daphnia were exposed to a single nominal concentration of 100 mg/L over a period of 48 hours. No effect (immobility) was observed up to the limit of water solubility. The EC50 (48 h) was determined as > 0.01 mg/L (measured initial concentration).

Based on the available results from structurally related read-across substances (in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5) which are characterized by a similar ecotoxicological profile and comparable structure, it can be concluded that Glycerol trioctanoate (CAS 538-23-8) shows no acute toxicity to aquatic invertebrates up to the limit of water solubility (< 0.532 µg/L at 20 °C).