reaction products of 1 mole of Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs and 1 mole of cyclohexylamine

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name PROCHINOR B657
CAS Number 2060540-82-9
Reference PROCHINOR B657
Batch number EP7377
Purity (%) 10
Expiry date 21/11/19

Analytical monitoring:

yes

Details on sampling:

Description of the protocol

For CHA: One to three steps of dilution into a 90:10 water:acetonitrile solution acidified with 0.1% formic acid or 100% acetonitrile with 0.2% formic acid, addition of an internal standard before filtration through a 0.2 µm x 13 mm PTFE filter into a vial for LC-MS/MS analysis. 5 µL were injected into the LC-MS/MS system.

For DDBS: One or two steps of dilution into a 30:70 water:acetonitrile solution acidified with 0.1% formic acid or 100% acetonitrile with 0.013% formic acid. 1 µL was injected into the LC-MS/MS system.

Vehicle:

no

Details on test solutions:

Test solution preparation

The test solutions were prepared by dilution of a primary stock solution or by directly dissolving the test item in the test solution


Treatment Control PROCHINOR B657
ID C06 C07 C08 C09 C10 C11 C14
Target rate [mg test item.L-1] NA 3.1 6.25 12.5 25 50 100
Actual amount of test item weighed for preparation of SS [mg] NA 51.2 51.2 51.2 51.2 51.2 54.7
Actual volume of SS [mL] NA 500 500 500 500 500 500
Actual volume sampled from SS [mL] NA 7.8 15.6 31.25 62.5 125 250
Final volume [mL] NA 250 250 250 250 250 250

SS= Stock Solution; NA= Not Applicable

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia, Daphnia magna Strauss clone 5, between 6 and 24 hours old and produced from parthenogenically reproducing brood female population were used. Daphnia are issued from the breeding lab. The selection of the juvenile was daily realized by filtration. The juvenile stemming at least from the second litter were used to start the test.

Culture conditions:
Medium: Elendt M4
Temperature: 20 °C ± 2
Light/dark cycle: 16 h/8 h
pH: Between 6 and 9
Hardness (CaCO3): Between 140 and 250 mg.L-1
Daily feeding: Pseudokirchneriella subcapitata and Chlorella vulgaris suspensions, spirulin

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.4 - 20.5 (°C)

pH:

8.0 - 8.2

Dissolved oxygen:

9.0 - 9.2 (mg O2/L)

Nominal and measured concentrations:

Nominal concentrations: 3.1 ; 6.25 ; 12.5 ; 25 ; 50 and 100 mg test item.L-1
Measured concentrations: 2.16 ; 3.02 ; 9.16 ; 18.36 ; 41.58 ; 90.99 mg test item. L-1

Details on test conditions:

Based on the results of the no-GLP range-finder, concentrations were prepared in OECD medium at 6 rates equivalent to: 3.1 ; 6.25 ; 12.5 ; 25 ; 50 and 100 mg test item.L-1. A control was set up with the test medium only. There were 4 replicates for each treatment and for the control. 5 daphnids were introduced in each replicate (each one containing 50 mL of test solution).
Organisms were maintained at temperature of 20°C ± 2°C and light intensity under 300 lux with a light:dark cycle of 16 hours light and 8 hours dark.
After 24 and 48 hours, the number of immobile daphnids in each replicates were recorded. pH dissolved oxygen and temperature were recorded at the beginning and the end of the test.

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

40.64 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

act. ingr.

Details on results:

cf Table 1

Results with reference substance (positive control):

24-hour EC50: 1.428 mg.L-1

cf details in Table 2

TABLE 1:

ID	Target rate	Immobilized daphnids 48 hours after exposure				Total of immobilized daphnids	Mean of immobilized daphnids (%)
	[mg test item.L^-1]	Replicates
		A	B	C	D
C06	0	0	0	0	0	0	0
C07	3.1	0	0	0	0	0	0
C08	6.25	0	0	0	0	0	0
C09	12.5	0	0	0	0	0	0
C10	25	1	0	0	0	1	5
C11	50	2	3	3	1	9	45
C14	100	5	5	5	5	20	100

TABLE 2:

ID	Rate	Immobilized daphnids 24 hours after exposure				Total of immobilized daphnids	Mean of immobilized daphnids (%)
	[mg reference item.L^-1]	Replicates
		A	B	C	D
T	0	0	0	0	0	0	0
C1	0,5	0	0	0	0	0	0
C2	0,9	0	1	0	0	1	5
C3	1,35	3	1	2	2	8	40
C4	2,025	5	4	4	5	18	90
C5	3,0375	5	5	5	5	20	100

Validity criteria fulfilled:

yes

Conclusions:

CONCLUSION

EC50-48h (95% confidence limits) 40.64 mg.L-1 (33.03– 49.83 mg.L-1)

The acute effects of PROCHINOR B657 diluted in OECD test medium at the definitive rates of 3.1 ; 6.25 ; 12.5 ; 25 ; 50 and 100 mg.L-1 to Daphnia magna were investigated under laboratory conditions.
The study is valid since
- immobilization in the control was less than 10% (actual values: 0% in the definitive test)
- dissolved oxygen in the lowest concentration producing the maximum effect at the end of the test was = 3 mg.L-1 (actual values: 9.1 mg.L-1 mg.L-1 in the definitive test)
- the EC50 with reference item over a period of 24 hours was between 0,6 and 2,1 mg.L-1 (actual value: 1.43 mg.L-1).

Executive summary:

The objective of this study was to assess the effects of the test item PROCHINOR B657 on the mobility of Daphnia magna over a period of 48 hours under static conditions according to OECD Guideline 202 (April 2004).

The test item is a soluble substance under the test conditions and the test solutions were prepared by dilution of a primary stock solution or by directly dissolving the test item in the test solution.

Test item quantification was performed according to a validated analytical method using a LC-MS/MS from Sciex (Exion LC and MS QTRAP 5500) as mass spectrometry detection to measure both DDBS and CHA from PROCHINOR B657 concentrations at 0 and 48 hours.

Results are the following:

	PROCHINOR B657
EC10-48h(95% confidence limits)	24.06 mg.L^-1(15.34 – 30.17 mg.L^-1)
EC50-48h(95% confidence limits)	40.64 mg.L^-1(33.03– 49.83 mg.L^-1)

EC = effect concentration

The acute effects of PROCHINOR B657 diluted in OECD test medium at the definitive rates of 3.1 ; 6.25 ; 12.5 ; 25 ; 50 and 100 mg.L^-1to Daphnia magna were investigated under laboratory conditions.

The study is valid since immobilization in the control was less than 10% (actual values: 0% in the definitive test), dissolved oxygen in the lowest concentration producing the maximum effect at the end of the test was = 3 mg.L^-1(actual value: 9.1 mg.L^-1in the definitive test) and the EC₅₀with reference item over a period of 24 hours was between 0,6 and 2,1 mg.L^-1(actual value: 1.43 mg.L^-1).

 

Description of key information

The objective of this valid study was to assess the effects of the test item on the mobility of Daphnia magna over a period of 48 hours under static conditions according to OECD Guideline 202 (April 2004).

The test item is a soluble substance under the test conditions and the test solutions were prepared by dilution of a primary stock solution or by directly dissolving the test item in the test solution.

Test item quantification was performed according to a validated analytical methodusing a LC-MS/MS from Sciex (Exion LC and MS QTRAP 5500) as mass spectrometry detection to measure both DDBS and CHA from PROCHINOR B657 concentrations at 0 and 48 hours.

The acute effects of the test item diluted in OECD test medium at the definitive rates of 3.1 ; 6.25 ; 12.5 ; 25 ; 50 and 100 mg.L^-1to Daphnia magna were investigated under laboratory conditions.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

40.64 mg/L