Propanoic acid, 2,2,3-trifluoro-3-[1,1,2,2,3,3-hexafluoro-3-(trifluoromethoxy)propoxy]-, methyl ester

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 September 2007 - 26 September 2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study conducted under GLP. Study on ammonium salt of hydrolysis product.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: ISO International Standard 10706:2000

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

ADONA

IUPAC Name:

ADONA

Details on test material:

- Name of test material (as cited in study report): MTDID 6675
- Substance type: single-constituent substance
- Physical state: clear colorless liquid
- Analytical purity: 29.9 ±0.1% in water
- Lot/batch no.: 140499-19/10
- Stability under test conditions: Stable in water at least 96 hours
- Storage condition of test material: At room temperature in the dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: 2 mL aliquots taken from all concentrations and control (spent solutions were pooled before sampling)
1st Study) Aliquots were taken on day 0, 2, 7, 9, 16, and 19 at the beginning and end of the intervals
2nd Study) Aliquots were taken on day 0, 3, 8, 10, 17, and 20 at the beginning and end of the intervals
- Sample storage conditions before analysis: Samples were stored in a freezer until analysis

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Method: ADONA consists of 29.9% active ingredient (a.i.) in water. The lower concentrations were prepared by subsequent dilutions of the highest test concentration (330 mg/L in test medium). No special treatment other than a 15 min of magnetic stirring was necessary to completely dissolve the test substance in the test medium.
- Controls: Test medium without test substance or other additives
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None, final test solutions were all clear and colorless.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Straus, 1820
- Source: In-house laboratory culture with known history
- Age of parental stock (mean and range, SD): No more than two weeks old; test organisms from second to 5th brood of parental stock
- Method of breeding: By acyclical parthenogenesis. Each batch started with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 L of M7 medium (modified ISO medium) in an all-glass culture vessel. Cultures maintained for a maximum age of 4 weeks. After 7 days of cultivation half of the M7 medium was renewed twice a week. Cultures were maintained at 18 - 22 °C
- Feeding during test: Yes
- Food type: Chlorella pyrenoidosa suspension
- Amount: 0.25 mL containing 1.3E+08 (1st study) or 1.8E+08 (2nd study) cells, corresponding to 0.2 mg carbon/daphnid/day.
- Frequency: Daily

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Remarks on exposure duration:

Two reproduction studies were preformed, the second study was done to confirm the results of the first study.

Test conditions

Hardness:

1st study (179 mg/L CaCO3); 2nd study (161−179 mg/L CaCO3)

Test temperature:

1st study (19.4 - 20.1 °C); 2nd study (18.7 - 19.5 °C)

pH:

1st study (7.2 - 8.9); 2nd study (7.4 - 9.0)

Dissolved oxygen:

1st study (6.8 - 10.6 mg/L); 2nd study (7.4 - 9.4 mg/L)

Nominal and measured concentrations:

[1st study (Nominal: 33, 59, 106, 185, and 330 mg/L, corresponding to: 10, 18, 32, 56, and 100 mg a.i./L); (Mean Measured: 9.6, 18.2, 32.2, 55.4, and 100.3 mg a.i./L)]
[2nd study (Nominal: 33, 106, and 330 mg/L, corresponding to 10, 32, and 100 mg a.i./L), (Mean Measured: 9.6, 32.8, and 102.2 mg a.i./L)]. Measured concentrations were stable and in agreement with nominal during the various periods of renewal (91 - 106% over both studies). Therefore, all results were based on nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: 60 mL all-glass vessel covered with a Perspex plate, 50 mL fill volume
- Aeration: Not reported
- Renewal rate of test solution (frequency/flow rate): Three times a week
- No. of organisms per vessel: 1 neonate, < 24 hours old
- No. of vessels per concentration (replicates): [1st study:10], [2nd study: 20]
- No. of vessels per control (replicates): [1st study: 20], [2nd study: 20]

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water used to prepare media was tap water purified by reverse-osmosis (RO-water, GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
- Culture medium different from test medium: No, M7 used throught the test. M7 is based on adjusted ISO medium supplemented with additional minerals.

OTHER TEST CONDITIONS

- Adjustment of pH: No
- Photoperiod: 16h light; 8h dark
- Light intensity: [1st study Start: 676−726 lux (9.4−10.0 µE.mˉ².sˉ¹); End: 617−661 lux (8.8−9.9 µE.mˉ².sˉ¹)], [2nd study Start: 719−780 lux (10.4−11.3 µE.mˉ².sˉ¹); End: 647−755 lux (9.8 - 10.3 µE.mˉ².sˉ¹)]

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : For the parental Daphnia, every workday and upon renewal on non-workdays the number of living, immobile and dead were recorded, along with the presence of eggs in brood pouch. Body length of the parental Daphnia was recorded at the end of the test. For the F1 generation, the number of newborn young and their condition was recorded every workday and upon renewal on non-workdays. Appearance of the first brood, presence of unhatched eggs and incidence of F1 generation immobility were recorded.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks on result:

other: ADONA did not adversely affect survival, growth, or reproduction of Daphnia magna at analytically confirmed nominal concentration of 100 mg a.i./L, corresponding to 330 mg/L of total product.

Details on results:

Exposure concentrations were based on nominal concentrations as analytical results showed that concentrations were within 80-120% of nominal. See Table 1 and 2.
- Mortality of parent animals: Controls < 20%, Treatment between 10 - 40% (not dose-responsive)
- Body length and weight of parent animals: No significant differences were found between controls and test samples with respect to surviving parental Daphnid length. See Table 4
- Time to first brood release or time to hatch: 8 days, See Table 3
- Effect concentrations exceeding solubility of substance in test medium: No

Reported statistics and error estimates:

Total reporduction on day 21 was tested for normality and for homogeneity of variance and statistically tested using an ANOVA test followed by a Bonferroni t-test and a Tukey test (TOXSTAT Release 3.5, 1996, D.D. Gulley, A.M. Boelter, H.L. Bergman).

Any other information on results incl. tables

Table 1, Measured Concentrations of ADONA in Test Samples (1^stStudy)^A
Sampling Time (Day)	Nominal Test Concentration (mg/L)	Measured Concentration (mg/L)	Relative to Nominal (%)	Relative to initial (%)
	0	<0.08
	10	9.43	94
0	18	18.0	100
(Fresh)	32	32.2	101
	56	55.3	99
	100	100	100
	0	<0.08
	10	9.65	97	102
2	18	18.2	101	101
(Old)	32	32.5	102	101
	56	55.1	98	100
	100	95.0	95	95
	0	<0.08
	10	9.53	95
7	18	17.5	97
(Fresh)	32	32.2	101
	56	54.7	98
	100	105	105
	0	<0.08
	10	9.09	91	95
9	18	17.9	99	102
(Old)	32	32.1	100	100
	56	53.5	96	98
	100	101	101	97
	0	<0.08
	10	9.86	99
16	18	18.7	104
(Fresh)	32	33.2	104
	56	57.5	103
	100	97.8	98
	0	<0.08
	10	9.83	98	100
19	18	18.8	104	100
(Old)	32	31.0	97	93
	56	56.4	101	98
	100	103	103	105
	0	<0.08
	10	9.6
Average	18	18.2
	32	32.2
	56	55.4
	100	100.3
A) Concentrations were corrected for the purity of the test substance (29.9%)

Table 2, Measured Concentrations of ADONA in Test Samples (2^ndStudy)^A
Sampling Time (Day)	Nominal Test Concentration (mg/L)	Measured Concentration (mg/L)	Relative to Nominal (%)	Relative to initial (%)
	0	<0.08
0	10	9.33	93
(Fresh)	32	31.8	99
	100	102	102
	0	<0.08
3	10	9.68	97	104
(Old)	32	32.3	101	102
	100	98	98	96
	0	<0.08
8	10	9.35	93
(Fresh)	32	32.8	103
	100	104	104
	0	<0.08
10	10	9.34	93	100
(Old)	32	32.9	103	100
	100	100	100	96
	0	<0.08
17	10	9.93	99
(Fresh)	32	33.8	106
	100	104	104
	0	<0.08
20	10	10.0	100	101
(Old)	32	33.3	104	98
	100	105	105	101
	0	<0.08
Average	10	9.6
	32	32.8
	100	102.2
A) Concentrations were corrected for the purity of the test substance (29.9%)

Table 3, Summary of Surviving First-Generation Daphnids
Nominal ADONA Concentration^A (mg/L)	Number exposed	First Day of Reproduction	Number of Surviving Adults	Mean Number of Live Young Per Surviving Adult ± Std. Dev.
1^stStudy
Negative control	20	8	18	136.1 ± 13.3
10	10	8	6	125.8 ± 23.2
18	10	8	9	158.4 ± 18.2
32	10	8	7	164.0 ± 26.5
56	10	8	7	174.2 ± 21.5
100	10	8	9	152.4 ± 39.5
2^ndStudy
Negative control	20	8	19	115.8 ± 26.6
10	20	8	16	136.7 ± 13.8
32	20	8	18	132.3 ± 34.5
100	20	8	20	142.5 ± 14.4
A) Concentration of active ingredient

Table 4, Summary of Length of parental Daphnids
Nominal ADONA Concentration (mg/L)	Number Exposed	Number of Surviving Adults	Mean Length ± Std. Dev. (mm)
1^stStudy
Blank control	20	18	4.2 ± 0.2
10	10	6	4.6 ± 0.2
18	10	9	4.7 ± 0.2
32	10	7	4.6 ± 0.3
56	10	7	4.7 ± 0.2
100	10	9	4.6 ± 0.2
2^ndStudy
Blank control	20	19	4.1 ± 0.1
10	20	16	4.2 ± 0.1
32	20	18	4.2 ± 0.1
100	20	20	4.2 ± 0.1
A) Concentration for active ingredient

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Parent animal (female Daphnia) mortality < 20% at test end. Average cumulative # of young/female in controls after 21 days ≥ 60 both studies.

Conclusions:

In two separate studies, the21-day NOEC of ADONA to Daphnia magna was 100 mg/L (OECD 211), based on the analytically confirmed nominal concentrations of the active ingredient.

Executive summary:

MeDONA is hydrolytically unstable. Aquatic toxicity testing on MeDONA would in practice be testing of the hydrolysis products. This test examined the ammonium salt of the hydrolysis product (ADONA).

ADONA did not adversely affect survival, growth, or reproduction of Daphnia magna, with a 21-day NOEC of 100 mg/L of active ingredient, which corresponds to 334 mg/L of total product.

The study was performed in accordance with internationally-accepted test guidelines and Good Laboratory Practice (GLP) standards.Therefore, this study is reliable without restrictions and the results are suitable for purposes of Risk Assessment, Classification & Labeling, and PBT Analysis.

Test Type: Semi-Static

21-d NOEC = 100 mg/L active ingredient

This corresponds to 334 mg/L total product