Glycerol trioctanoate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 Jan-21 Feb 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Qualifier:

according to guideline

Guideline:

EU Method C.20 (Daphnia magna Reproduction Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, Düsseldorf, Germany

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A concentrated stock solution of the test item was prepared by weighing 200 mg of the test item onto a small Teflon plate. The Teflon plate was placed inside a sterile 2L brown glass Schott bottle with drain port. Dilution water was then added to a volume of 2 L and the solution stirred at 300 rpm for 24 hours at room temperature (21 ± 2°C). The WAF solution was then left to settle for 24 hours before the first 100 mL was drained and the remaining solution was contained in a sterile Schott bottle and used to prepare the test concentrations. The WAF solution formed the test loading of 1 mg/L.
- Controls: yes, blank control
- Evidence of undissolved material: none reported

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Straus
- Source: German Federal Environment Agency, Institute for Water, Soil, and Air Hygiene (Berlin).
- Feeding during test: The daphnids were fed concentrated suspensions of living cells of the unicellular alga Desmodesmus subspicatus. Feeding occurred at each media renewal. The algae were spun at 3000 rpm for 3 min at room temperature, the supernatant discarded and the optical density at 585 nm measured from a diluted sample. The algal feed concentrate was stored at 4°C and kept for a maximum of one week before replacing it with a fresh suspension. The required aliquot of the algal concentrate per test vessel was calculated from a calibrated nomograph. Each daphnid was fed daily with a volume of algae containing 0.2 mg carbon, independent of age.

ACCLIMATION
- Acclimation period: For use in the test, newborn Daphnia (between 4 and 24 h old) were removed by wide-bore pipette (to avoid damage) and isolated in fresh dilution water for at least 1 h prior to being added to the test vessels.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

1 mmol/L

Test temperature:

18.5-19.9 °C

pH:

7.70-9.61

Dissolved oxygen:

6.5-9.8 mg O2/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal: 1 mg/L (limit test)

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass flasks stoppered with silicon plugs
- Renewal rate of test solution (frequency/flow rate): test solutions were renewed daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Purified local tap water will be used in accordance with the OECD guideline. The tap water is sourced from the Schmallenberg district water production plants, mostly fed by small springs and percolation. The purification process occurs on site at Fraunhofer IME and includes filtration with activated charcoal, passage through a lime-stone column, and aeration to the point of oxygen saturation. To avoid copper contamination (as copper is toxic to aquatic organisms), plastic water pipes are used in the test facilities.
- Particulate matter:
- Metals: Cu: 0.880-<0.943 µg/L
Fe: 5.54 µg/L
Mn: 1.49-2.03 µg/L
Zn: 2.62-2.36 µg/L
- Chlorine: < 0.02 mg/L
- Ca/mg ratio: 0.7/0.3 mmol/L
- Conductivity: 216.8 µS/cm

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light/8 hours darkness
- Light intensity: 609-966 lux

EFFECT PARAMETERS MEASURED: The number of juvenile Daphnia magna per replicate was recorded daily from day 6 onwards. Immobility, abnormalities in appearance, behaviour, condition (including male sex) or presence of winter eggs (ephippium) were also recorded. At study termination, body length (excluding the anal spine) of the adults in all treatments without significant immobility was measured by digital photography. Image analysis allowed comparison of the treatment group daphnids with the equally measured control daphnids.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Details on results:

- The cumulative number of offspring per replicate of the control at test end was 65 for the control and 69 for the test loading. No significant reproduction inhibition effect was shown so the NOELR for reproduction is ≥ 1.0 mg/L.

- No immobilisation occurred throughout the test. No other clinical symptoms were observed. The NOELR for immobility is ≥ 1.0 mg/L.

- Adult growth (length) was not affected by the test loading. The NOELR for length is ≥ 1.0 mg/L.

- Mean age of first brood was day 11.1 and 10.2 for the test loading and the control, respectively. A significant difference between the control and 1.0 mg/L was shown (p=0.019). The NOELR could not be determined for this parameter.

Reported statistics and error estimates:

- The data were tested for variance homogeneity and normal distribution. The NOELR was calculated by Students T-Test. The NOELR was reported as more than the highest concentration tested when the statistical test failed to find a difference between the limit treatment and the control. All statistical evaluations were conducted using the computer software ToxRat® Professional v. 2.10.

Table1. Survival, growth and reproduction averages for all endpoints of Daphnia magna at the end of the 21 d exposure period to the test item compared to the control treatment.

Nominal treatment loading	Parental immobility		Growth (length on day 21)	Age at first brood	Cumulative offspring per female
(mg/L)	(%)		Mean ± SD (mm)	Mean ± SD (days)	Mean ± SD (n)
Control	0		3.98 ± 0.26	10.2 ± 0.95	64.7 ± 12.4
1.0	0		3.97 ± 0.25	11.1 ± 0.84	69 ± 7.4
NOEL	≥1.0 mg/L	≥ 1.0 mg/L		n.d.	≥ 1.0 mg/L

n.d.: not able to be determined from the data as there were effects at the response level.

 

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 Feb - 21 March 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: OECD guideline 202 Part II 1993, which is now OECD guideline 211

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Samples were taken after 0, 24, 48 and 72 h from both concentrations

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Two stock solutions with nominal concentrations of 1 and 100 mg/L test substance were prepared in M4-medium. They were stirred for at least 16h and up to 7 days. These suspensions were carefully filtered with a glass fibre filter (pretreated with NaOH). These solutions saturated with test substance were used without further dilution for the test

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: STRAUS 1820
- Source: BGA Berlin (May 1994), afterwards own breeding
- Feeding during test
- Food type: Scenedesmus subspicatus
- Amount: 0-7 days old: 2.5x10E6 cells/daphnid/d; > 7 days old animals: 5.0X10E6 cells/daphnid/d

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Test temperature:

20.2 - 20.6 °C

pH:

Control:7.5 - 7.9
1 mg/L: 7.7 - 8.0
100 mg/L: 7.6 - 7.8

Dissolved oxygen:

Control: 93 - 100%
1 mg/L and 100 mg/L: 92 - 100%

Nominal and measured concentrations:

Nominal concentrations: 1 and 100 mg/L
Measured concentrations: 1 mg/L: < 0.05 mg/L; 100 mg/L: < 0.05 mg/L, the test solution prepared at 11.3.1996 had 2.57 mg/L. The analytical recovery was partly above the water solubility determined with the column elution procedure. Thus, a complete separation of undissolved test substance was not always obtained.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL Erlenmeyer flasks, filled with 50 mL, covered with a watch glass
- Renewal rate of test solution (frequency/flow rate): every 2-3 days
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10


TEST MEDIUM / WATER PARAMETERS
- Medium: M4-Medium according to Elendt


OTHER TEST CONDITIONS
- Photoperiod: 16 hours
- Light intensity: 300 Lux


EFFECT PARAMETERS MEASURED: Immobility and reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

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

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

immobilisation

Remarks on result:

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

Details on results:

- Immobilisation of parent animals: One in the control, one at 1 mg/L
- No. of offspring produced per day per female: see tables 1-4
- Time to first brood release or time to hatch: First offspring 9 days after test start.

Reported statistics and error estimates:

T-Test, Two Sample F-Test

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

Table 1: Reproduction rate: Number of offspring produced per daphnia alive per day (calculated from the first day of occurrence of offspring in the control)

Concentration	Replicates										Offspring/ Daphnia/d [mean value]
	1	2	3	4	5	6	7	8	9	10
0 mg/L	5.0	4.5	4.6	5.8	4.6	4.9	0.0	4.9	4.4	5.0	4.9
1 mg/L	3.4	3.1	5.6	2.5	3.8	3.6	6.1	0.0	4.2	5.0	4.1
100 mg/L	5.0	4.8	4.5	6.2	3.3	3.2	4.1	4.1	1.6	3.6	4.0

 

Table 2: Control: Number of offspring on different days

	Offspring after x days									Offspring (sum)	Offspring/ Daphnia/d
	2	5	7	9	12	14	16	19	21(22)
Parallel 1	-	-	-	-	14	-	18	16	17	65	5.0
Parallel 2	-	-	-	-	11	-	20	17	10	58	4.5
Parallel 3	-	-	-	-	14	20	-	10	16	60	4.6
Parallel 4	-	-	-	8	16	-	18	20	13	75	5.8
Parallel 5	-	-	-	-	14	18	-	13	15	60	4.6
Parallel 6	-	-	-	-	15	-	17	20	12	64	4.9
Parallel 7	-	-	T
Parallel 8	-	-	-	-	15	-	22	16	11	64	4.9
Parallel 9	-	-	-	-	5	20	-	17	15	57	4.4
Parallel 10	-	-	-	-	10	19	-	16	20	65	5.0
							Mean offspring:			63	4.9

T) Immobilisation of parental animal

 

Table 3: Test concentration 1 mg/L: Number of offspring on different days

	Offspring after x days									Offspring (sum)	Offspring/ Daphnia/d
	2	5	7	9	12	14	16	19	21(22)
Parallel 1	-	-	-	7	1	10	-	7	19	44	3.4
Parallel 2	-	-	-	-	-	13	19	8	-	40	3.1
Parallel 3	-	-	-	9	11	-	18	19	16	73	5.6
Parallel 4	-	-	-	-	-	-	7	14	12	33	2.5
Parallel 5	-	-	-	-	3	14	-	18	15	50	3.8
Parallel 6	-	-	-	-	8	17	-	9	13	47	3.6
Parallel 7	-	-	-	-	9	18	20	19	13	79	6.1
Parallel 8	-	-	T
Parallel 9	-	-	-	-	8	18	-	15	13	54	4.2
Parallel 10	-	-	-	-	15	-	19	16	15	65	5.0
							Mean offspring:			54	4.1

T) Immobilisation of parental animal

Table 4: Test concentration 10 mg/L: Number of offspring on different days

	Offspring after x days									Offspring (sum)	Offspring/ Daphnia/d
	2	5	7	9	12	14	16	19	21(22)
Parallel 1	-	-	-	-	8	22	-	21	14	65	5.0
Parallel 2	-	-	-	-	10	21	-	18	13	62	4.8
Parallel 3	-	-	-	-	8	20	-	14	16	58	4.5
Parallel 4	-	-	-	-	13	10	23	14	20	80	6.2
Parallel 5	-	-	-	-	-	3	16	24	-	43	3.3
Parallel 6	-	-	-	-	-	6	18	17	-	41	3.2
Parallel 7	-	-	-	-	9	15	-	12	17	53	4.1
Parallel 8	-	-	-	-	5	-	12	21	15	53	4.1
Parallel 9	-	-	-	-	-	3	-	7	11	21	1.6
Parallel 10	-	-	-	-	-	-	11	20	16	47	3.6
							Mean offspring:			52.3	4.0

Reference 3

Endpoint:

long-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

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: Source: CAS 620-67-7, Oleon, 2013, Daphnia magna

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

immobilisation

Remarks on result:

other: Source: CAS 620-67-7, Oleon, 2013, Daphnia magna

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks on result:

other: Source: CAS 620-67-7, Oleon, 2013, Daphnia magna

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: Source: CAS 65381-09-1, BASF, 1996, Daphnia magna

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

immobilisation

Remarks on result:

other: Source: CAS 65381-09-1, BASF, 1996, Daphnia magna

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: Source: CAS 65381-09-1, BASF, 1996, Daphnia magna

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

immobilisation

Remarks on result:

other: Source: CAS 65381-09-1, BASF, 1996, Daphnia magna

Additional information

Since no studies investigating the long-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), 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 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 as a limit test in a semi-static design and under GLP according to OECD testing guideline No. 211 (Daphnia magna Reproduction Test) and EU Method C.20 (Daphnia magna Reproduction Test). The freshwater flea Daphnia magna was used as test organism. The daphnia were exposed to a single nominal concentration of 1 mg/L over a period of 21 days with daily renewal of the test solutions. No effect (immobility) was observed in the test solutions. No significant difference was found between the control and test solutions for growth rate and offspring count. The NOELR (21d) was determined as≥1 mg/L for immobility, growth rate and offspring count.

The study with read-across substance Glycerides, C8-10 mono and di (CAS 85536-07-8) was performed in a semi-static design under GLP (with acceptable restrictions) according to OECD guideline 202 Part II (1993), which is now OECD guideline 211. The freshwater flea Daphnia magna was used as test organism. The daphnia were exposed to nominal concentrations of 1 and 100 mg/L (measured: < 0.05 mg/L WAF for both test concentrations) over a period of 21 days. No effect (immobility, reproduction) was observed up to the limit of water solubility. The NOEC (21 d, immobility, reproduction) was determined as≥0.01 mg/L. The LOEC (21 d, immobility, reproduction) was determined as > 0.01 mg/L.

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 long-term toxicity to aquatic invertebrates up to the limit of water solubility (< 0.532 µg/L).