(2S)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-amine hydrochloride (1:1)

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 Aug 2017 - 19 Sep 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(2S)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-aminium chloride

EC Number:

695-797-0

Cas Number:

93601-86-6

Molecular formula:

C14H21NO.HCl

IUPAC Name:

(2S)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-aminium chloride

Test material form:

solid: particulate/powder

Details on test material:

- Appearance: White to off-white powder
- Storage condition of test material: At room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and all test concentrations
- Sampling method: 4.0 mL taken from the approximate centre of the test vessels, at t=0 h and t=48 h.
- At the end of the exposure period, the replicates were pooled at each concentration before sampling.
- Sample storage conditions before analysis: Samples were stored in a freezer (≤-15°C) until analysis at the analytical laboratory of the Test Facility.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Direct addition to the test medium. Test solutions were prepared at the highest concentration of 100 mg/L applying 15-26 minutes of magnetic stirring to accelerate dissolution of the test item in medium. Lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium. All test solutions were clear and colorless at the end of the preparation procedure.
- Controls: Test medium without test item or other additives.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Straus 1820
- Source: In-house laboratory culture with a known history
- Age of parental stock: >2 weeks old
- Age at test initiation: <24 hours
- Validity of batch: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

CULTIVATION
- Start of each batch: With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of M7 medium in an all-glass culture vessel.
- Maximum age of the cultures: 4 weeks
- Renewal of the cultures: After 7 days of cultivation half of the medium twice a week.
- Temperature of medium: 18-22°C
- Feeding: Daily, a suspension of fresh water algae.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

20-21°C

pH:

7.7-7.9

Dissolved oxygen:

8.5-9.0 mg/L

Nominal and measured concentrations:

Nominal: 10, 18, 32, 56, 100 mg/L
Measured concentrations were 108-117% of nominal throughout the test. Therefore, effect parameters were based on nominal concentrations. See Table 1 in 'Any other information on results incl. tables' for details on measured concentrations throughout the test.

Details on test conditions:

TEST SYSTEM
- Test vessel: 60 mL all-glass, containing 50 mL of test solution
- Aeration: no
- Feeding: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium, prepared with tap water purified by Reverse Osmosis
- Culture medium different from test medium: yes, M7 medium
- Intervals of water quality measurement: Dissolved oxygen and pH: at beginning and end of the test, for all concentrations and the control. Temperature: continuously in a temperature control vessel, beginning at the start of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h photoperiod daily

EFFECT PARAMETERS MEASURED
- Immobility (including mortality) at 24 hours and at 48 hours.

RANGE-FINDING STUDY
- Test concentrations: Control, 0.10, 1.0, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes; expected EC50 was between 10 and 100 mg/L (nominal concentration).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (performed Sep 2017)

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

33 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% c.i.: 27-40 mg/L

Details on results:

- Behavioural abnormalities: none observed in the Control group.
- Mortality of control: none
- All water quality parameters remained within the requirements as laid down in the study plan throughout the test.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- The 24h-EC50 was 0.59 mg/L with a 95% confidence interval between 0.52 and 0.68 mg/L.
- The 48h-EC50 was 0.41 mg/L with a 95% confidence interval between 0.34 and 0.46 mg/L.
- The actual responses in this reference test with K2Cr2O7 are within the ranges of the expected responses at the different concentrations, i.e. the 48h-EC50 was between 0.3 and 1.0 mg/L. Hence, the sensitivity of this batch of D. magna was in agreement with the historical data collected at the Test Facility.

Reported statistics and error estimates:

24 and 48h-EC50-values were calculated using the Spearman-Karber method (without trim) on the percentages of affected daphnids and the logarithms of the corresponding test item concentrations.
ToxRat Professional v 3.2.1 (ToxRat Solutions® GmbH, Germany) was used to perform the analyses.

Any other information on results incl. tables

Table 1: Final Test: Test Samples

Time of sampling [hours]	Concentration [mg/L]		Relative to nominal [%]	Relative to initial [%]
	Nominal	Analyzed
0	0	n.d.	n.a.
	10	11.7	117
	18	20.4	114
	32	37.3	116
	56	61.7	110
	100	111	111
48	0	n.d.	n.a.	n.a.
	10	11.6	116	99
	18	20.2	112	99
	32	36.3	114	97
	56	60.7	108	98
	100	110	110	99

Samples were stored in the freezer (≤ -15°C) until the day of analysis.

n.d. Not detected.

n.a. Not applicable.

Table 2: Number of Introduced Daphnids and Incidence of Immobility in the Final Test

Time (h)	Replicate	Nominal conc. (mg/L)
		Control	10	18	32	56	100
0	A	5	5	5	5	5	5
	B	5	5	5	5	5	5
	C	5	5	5	5	5	5
	D	5	5	5	5	5	5
	Total introduced	20	20	20	20	20	20
24	A	0	0	0	0	0	3
	B	0	0	0	0	0	5
	C	0	0	0	0	0	4
	D	0	0	0	0	0	2
	Total immobilised	0	0	0	0	0	14
	Effect %	0	0	0	0	0	70
48	A	0	2	0	2	4	5
	B	0	0	1	3	5	5
	C	0	0	0	2	4	5
	D	0	0	0	3	3	5
	Total immobilised	0	2	1	10	16	20
	Effect %	0	10	5	50	80	100

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For details on validity criteria, please refer to section 'Overall remarks'

Conclusions:

The 48h-EC50 for Daphnia magna exposed to the test item was 33 mg/L, based on analytically confirmed nominal concentrations (95% confidence interval between 27 and 40 mg/L).

Executive summary:

In a 48 h acute toxicity test conducted according to OECD guideline 202, Daphnia magna were exposed to the test item at nominal concentrations of 10, 18, 32, 56 and 100 mg/L and an untreated control (all with 4 replicates per concentration and 5 daphnids per replicate). Measured concentrations were at 108 -117% relative to nominal throughout the test. Therefore, effect parameters were based on nominal concentrations. A dose-response relationship for immobility was observed at 48 h. The 48h-EC50 for Daphnia magna exposed to the test item was 33 mg/L, based on analytically confirmed nominal concentrations. The study is considered to be reliable without restrictions.