.alpha.-D-Glucopyranosyl bromide, 2,3,4,6-tetrakis(2,2-dimethylpropanoate)

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 May 2020 - 17 Aug 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2012

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MEP, PR China, the Guideline for the testing of chemicals, effects on biotic systems (2nd edition), No. 211 "Daphnia magna reproduction test", China Environment Press

Version / remarks:

2013.8

Qualifier:

according to guideline

Guideline:

other: Guidance document on analytical quality control and validation procedures for pesticides residues analysis in food and feed, SANCO/12571/2013

Version / remarks:

19 November 2013, implemented by 01/01/2014

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number: from sponsor, batch no: M19IB2844
- Purity: 100.3%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature (15-25 °C)
- Solubility of the test material in the solvent/vehicle and the exposure medium: solubility in water < 0.02 mg/L (20°C)

Analytical monitoring:

yes

Details on sampling:

- Concentrations of the test chemical were analyzed once a week in fresh and old media: 0, 8, 16d of fresh media, and 2, 10, 18d of old media
- 150 mL solution was sampled from each test concentration and control
- Pre-treatment method: 50 mL test samples were extracted with 2.0 mL n-hexane, after shaken for 30 seconds and the n-hexane phase was analyzed by GC/MS/MS. If necessary, the samples were further diluted 5-fold with n-hexane to obtain concentrations within the calibration range.
- Sample storage conditions before analysis: the extracted samples were stored in refrigerator (2-8 °C) until analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a concentration of 100 mg/L (100% SS) was prepared by adding a weighed amount of 0.20 g test item directly in 2 L test water, which was then stirred for 24 hours at about 400 rpm to ensure maximum dissolution in the test medium. The solution was filtered over a 0.45 µm filter after stirring and standing for about 20 min, to obtain a saturated solution. The filtered solution was diluted step-by-step to get the test solutions of 6.25 - 12.5 - 25 - 50 % of SS at 100 mg/L
- Controls: a concurrent control group was included in parallel with ca. 100 mL of the test water in each vessel
- Test concentration separation factor: 2

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Straus
- pre-cultured animals were maintained under culture conditions similar to those that were used in the test. Test animals were less than 24 hours old and not the first brood progency
- Source: Applied Chemistry Department Pesticide Environmental Evaluation Laboratory of China Agricultural University
- Feeding during test: yes
- Food type: living algal cells of Chlorella sp and Desmodesmus subspicatus
- Amount: 0.1-0.2 mg C/Daphnia/day
- Frequency: daily

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

140-250 mg/L CaCO3

Test temperature:

18-22 °C

pH:

6-8.5

Dissolved oxygen:

60% of air saturation value

Nominal and measured concentrations:

Nominal: 6.25 - 12.5 - 25 - 50 - 100% of SS at 100 mg/L
Geometric mean measured in fresh media: 0.0040 - 0.0099 - 0.0277 - 0.0488 - 0.1304 mg/L
Geometric mean measured in old media: 0.0037 - 0.0097 - 0.0274 - 0.0258 - 0.1310 mg/L.

The overall mean measured concentrations are 0.0039 - 0.0098 - 0.0276 - 0.0508 - 0.1307 mg/L, measured after 0d (fresh), 2d (old), 8d (fresh), 10d (old), 16d (fresh), 18d (old).

Details on test conditions:

TEST SYSTEM
- Test vessel: 200 or 250 mL glass beaker
- Test solution volume: 100 mL per vessel
- Aeration: before usage, the water was aerated for more than 24 hours to attain DO saturation of at least 80% ASV
- semi-static
- Renewal rate of test solution: every 48 hours
- 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: tap water treated with Aquapro Pure Water System
- Total organic carbon: < 2 mg/L
- characteristics of test water quality: in accordance with the GB11604-89 standard "water quality standard for fisheries"

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light / 8 hours dark
- Light intensity: 100-1500 lux for cool white light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- offspring (including presence of aborted eggs or dead offspring): daily
- mortality: daily
- number of living offspring produced per parent
- body length: at end of the test
- time to production of first brood, total juveniles of each parent, total broods of each parent

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: yes
- Test concentrations range-finding study: 0.8 - 4.0 - 20 - 100% of the saturated solution (SS) at 100 mg/L (nominal concentration)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.028 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality: Total number living offspring produced at the end of the test per parent daphnia at the start of the test excluding from the analysis parental accidental and/or inadvertent mortality {TG 211}

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.051 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality: Total number living offspring produced at the end of the test per parent daphnia at the start of the test excluding from the analysis parental accidental and/or inadvertent mortality {TG 211}

Details on results:

- Body length of parents:
NOEC 0.0276 mg/L
LOEC 0.0508 mg/L

- Mortality of parents:
NOEC 0.0508 mg/L
LOEC 0.1307 mg/L

- First brood time:
NOEC > 0.1307 mg/L
LOEC > 0.1307 mg/L

- Total number of broods per parent:
NOEC 0.0508 mg/L
LOEC 0.1307 mg/L

Reported statistics and error estimates:

- Mortality: no significant difference for mortality between control and the test groups of 6.25-12.5-25-50% of SS treatment groups (Student's-T, P>0.05), highly significant difference between control and 100% SS test group (Student's-T, P<0.001)
- Total living offspring: no significant difference between control and test groups 6.25-12.5-25%SS (student's T-test, P>0.05), highly significant difference between control and test groups 50-100% of SS (Student's-T, P<0.001). For the control group, the coefficient of variation was around the mean number of living offspring produced per parent animal at the end of the test was 24.0%
- First time brood: no significant difference between the control and all test groups (Student's-T, P>0.05)
- Body length: highly significant difference between control and test group of 50% and 100% of SS treatment groups (Student's-T, P<0.001)
- Total number of broods: no significant difference between the control and all test groups (Student's-T, P>0.05) except the 100%SS treatment group (Student's-T, P<0.001)

Accumulated mortality and observed toxicity signs

Nominal concentration	Maternal mortality (%)	Mean +- SD (%)	P-value (two sided)
control	0	NA	NA
6.25% SS	0	NA	NA
12.5% SS	0	NA	NA
25% SS	0	NA	NA
50% SS	20	20 +- 42	0.17
100% SS	80	80 +- 42	0.0002

 

Total juveniles per parent during the test

Nominal concentration	Total juveniles per parent, mean +- SD	CV (%)	P-value (two sided)	Inhibition rate (%)
control	145 +- 35	24.0	/	/
6.25% SS	140 +- 30	21.6	0.73	3.4
12.5% SS	149 +- 41	27.7	0.61	-2.1
25% SS	128 +- 33	26.0	0.40	11.7
50% SS	84 +- 44	52.7	0.004	42.1
100% SS	17 +- 33	200.1	0.00000007	88.3

 

First brood time of each parent

Nominal concentration	First brood time of each parent (day after exposure), mean +- SD	P-value (two sided)
control	9.4 +- 1.1	/
6.25% SS	10.4 +- 1.2	0.06
12.5% SS	10.2 +- 1.8	0.23
25% SS	10.6 +- 1.6	0.06
50% SS	9.4 +- 1.3	1.00
100% SS	9.8 +- 1.0	0.18

 

Body length of each parent

Nominal concentration	Body length (mm), mean +- SD	P-value (two sided)
control	2.88 +- 0.12	/
6.25% SS	2.90 +- 0.16	0.65
12.5% SS	2.89 +- 0.17	0.82
25% SS	2.79 +- 0.18	0.25
50% SS	2.68 +- 0.07	0.0008
100% SS	2.26 +- 0.02	0.000003

 

Total number of broods per parent

Nominal concentration	Total number of broods per parent, mean +- SD	P-value (two sided)
control	5.8 +- 0.9	/
6.25% SS	5.2 +- 0.8	0.13
12.5% SS	5.5 +- 1.1	0.51
25% SS	5.1 +- 0.7	0.08
50% SS	5.0 +- 1.2	0.12
100% SS	1.3 +- 1.8	0.000002

 

Water quality and physicochemical measurements in the definitive test

Nominal concentration	Measured item	2020-05-20	2020-05-22	2020-05-28	2020-05-30	2020-06-05	2020-06-07
		Fresh	Old	Fresh	Old	Fresh	Old
Control	pH	8.43	8.57	8.37	8.55	8.01	8.04
	D.O. (mg/L)	8.66	8.67	8.48	9.10	8.34	9.07
	Water hardness (CaCO3 mg/L)	180	-	180	-	200	-
6.25% SS	pH	8.41	8.57	8.12	8.54	8.12	8.43
	D.O. (mg/L)	8.68	8.46	8.34	9.10	8.23	8.86
12.5% SS	pH	8.48	8.56	8.18	8.58	8.18	8.49
	D.O. (mg/L)	8.60	8.49	8.27	9.07	8.15	9.09
25% SS	pH	8.54	8.55	8.32	8.58	8.18	8.50
	D.O. (mg/L)	8.60	8.50	8.20	9.06	8.10	9.14
50% SS	pH	8.51	8.53	8.42	8.51	8.19	8.49
	D.O. (mg/L)	8.61	8.46	8.22	9.04	8.04	9.11
100% SS	pH	8.53	8.54	8.47	8.55	8.22	8.45
	D.O. (mg/L)	8.50	8.49	8.26	9.06	8.13	9.14

 

Concentrations of test item in the exposure solution

Nominal concentration	Geometric mean concentration in fresh media (mg/L)	Geometric men concentration in old media (mg/L)	Overall mean concentration (mg/L)
6.25% SS	0.0040	0.0037	0.0039
12.5% SS	0.0099	0.0097	0.0098
25% SS	0.0277	0.0274	0.0276
50% SS	0.0488	0.0528	0.0508
100% SS	0.1304	0.1310	0.1307

 

 

Validity criteria fulfilled:

yes

Conclusions:

The objective of the test was to evaluate the effects of JNJ-42808389-AAA (T003421) on the
reproductive capacity output of Daphnia magna. For this purpose, test organisms were exposed to aqueous solutions containing the test item at various concentrations: 6.25 - 12.5 - 25 - 50 and 100% of saturated solution at 100 mg/L (nominal). Total number of living offspring per parent, body length of parent, maternal mortality, first brood time and total number of broods per parent were analyzed to obtain the NOEC and LOEC.

In conclusion, the 21-d NOEC and LOEC for all the parameters based on the measured concentration was: total number of living offspring: NOEC 0.0276 mg/L, LOEC 0.0508 mg/L. Body length: NOEC 0.0276 mg/L, LOEC 0.0508 mg/L. Mortality: NOEC 0.0508 mg/L, LOEC 0.1307 mg/L. First brood time: NOEC > 0.1307 mg/L, LOEC > 0.1307 mg/L. Total number of broods per parent: NOEC 0.0508 mg/L, LOEC 0.1307 mg/L. The results of the test can be considered reliable without restriction.

Description of key information

One key study (Hairong, 2021) is used for endpoint coverage. The study investigates the chronic toxicity of T003421 to Daphnia magna in a 21-day reproduction test according to the OECD guideline 211. The study determined a 21d-NOEC = 0.0276 mg/L (mean measured concentration). The study was assigned a Klimisch score of 1 and was conducted under GLP.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Effect concentration:

0.028 mg/L