L-Glutamic acid, N-acetyl-, compd. with (.alpha.S)-.alpha.-(2-methylpropyl)-2-(1-piperidinyl)benzenemethanamine (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:

22 January - 9 April 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to an appropriate EU test method and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Test guideline 3

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(S)-3-Methyl-1-(2-piperidine-1-phenyl)butylamine compound with N-Acetyl-L-glutamic acid

EC Number:

606-883-4

Cas Number:

219921-94-5

Molecular formula:

C16-H26-N2 x C7-H11-N-O5

IUPAC Name:

(S)-3-Methyl-1-(2-piperidine-1-phenyl)butylamine compound with N-Acetyl-L-glutamic acid

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): AGEE 623 Glutaminate; L-Glutamic acid, N-acetyl-, compd. with (S)-alpha-(2-methylpropyl-)-2-(1-piperidinyl)-benzenemethanamine
- Molecular formula (if other than submission substance): C16H26N2 * C7H11NO5
- Molecular weight (if other than submission substance): 435.57 g/mol
- Structural formula attached as image file (if other than submission substance): see reference substance
- Physical state: solid white powder
- Analytical purity: 100% (4% of (R)-(-)-enantiomer)
- Purity test date: 28 october 1998
- Lot/batch No.: T29725
- Expiration date of the lot/batch: 31 december 1999
- Stability under test conditions:pure see expiry date; in water: for at least 2 hours
- Storage condition of test material: in original container, at room temperature, in the dark

Test solutions

Vehicle:

no

Details on test solutions:

A stock solution of nominal 100 mg/L was prepared by dissolving 50 mg test substance in 500 ml test water. This stock solution was stirred on a magnetic stirrer for about 10 minutes. The stock solution was diluted in a series of sequential dilutions with test water to prepare the test media. The
test media were freshly prepared just before introduction of start of the test.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Straus), clone 5
- Source: supplied 1997 by the Umweltbundesamt, Institut für Wasser-, Boden- und Lufthygiene, Berlin, Germany
- Age at study initiation (mean and range, SD): 6 - 22 hours old
- Method of breeding: The Daphnia are bred in the laboratories of IBACON under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test. The test organisms were not first brood progeny.
- Sex: female

ACCLIMATION
- Acclimation period: for approximately 6 hours under test conditions
- Acclimation conditions (same as test or not): yes

Study design

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Test conditions

Hardness:

2.5 mmol/L (=250.0 mg/L) as CaCO3

Test temperature:

20 to 22°C

pH:

7.3 to 7.7

Dissolved oxygen:

at least 7.6 mg/L

Salinity:

CaCI2 x 2H20: 2.0 mmol/L (= 294.0 mg/L)
MgS04 X 7H20: 0.5 mmol/L (= 123.0 mg/L)
NaHC03: 0.75 mmol/L (= 65.0 mg/L)
KCI: 0.075 mmol/L (= 5.8 mg/L)

Nominal and measured concentrations:

nominal concentration stocksolution: 100 mg/l
nominal concentrations test solution: 0.10 mg/L, 0.21 mg/L, 0.46 mg/L, 1.0 mg/L, 2.1 mg/L and 4.6 mg/L
measured concentrations (mg/L): The analytically determined mean test substance concentrations in the analysed test media varied in the range
from 96% to 102% of the nominal values. In the stock solution sample 94% of the nominal concentration was measured.

Details on test conditions:

TEST SYSTEM
The Test vessels were glass beakers with 50 ml test medium. Each concentration had 4 replicate vessels; 20 individuals per control and concentration, divided into two groups of ten animals, each group in ml test medium. The age of the test organisms were < 24 hours and they were not fed during the test. Vessels were kept at 19.4 to 20.0°C .

The dilution water was synthetic freshwater with a hardness of 250.0 mg/L CaCO3.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

2.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Any other information on results incl. tables

After 24 hours in the control and in the test concentrations up to 2.1 mg test substance/L no mortality or immobilization of the test animals was observed. At the highest test concentration of 4.6 mg/L 20% of the Daphnia were immobile after 24 hours test duration.

The 24 hour EC50 and EC1000 of the test substance were clearly higher than 4.6 mg test substance/L. The 24 -hour EC0 was 2.1 test substance/L.

After 48 hours of exposure the toxicity of the test substance to Daphnia magna had increased. In the control and up to and including the test concentration of nominal 2.1 mg test substance/L no significant immobility or morality of the test animals or other signs of intoxication were determined during the test period of 48 hours. At the test concentration of nominal 0.46 mg/L one test animal was immobile at the observation after 48 hours. However, this immobilization rate was not estimated as a significant toxic effect, because according to the test guidelines this immobilization rate is also tolerated in the control, and additionally at the next higher test substance concentration of 1.0 mg test substance/L no immobilization and no other signs of intoxication were observed. At the test concentration of 2.1 mg/L no Daphnia were immobile, but all Daphnia were colourless. This effect might be caused by a toxic effect of the test substance.

The 48 hour EC50 was calculated to be 3.1 mg test substance/L with 95% confidence limits from 2.1 to 4.6 mg/L. The 48 hour EC0 was determined to be 2.1 mg/L. The 48 hour NOEC of AGEE 623 Glutaminate was 1.0 mg test substance/L, since no significant immobilization rate and no other signs of intoxication were observed at the test animals up to and including this test concentration. The 48 hour EC100 amounted to 4.6 mg test substance/L.

Biological results:

	Mean measured test concentration [mg/L]	lower 95% confidence value [mg/L]	upper 95% confidence value [mg/L]
24 hour EC50	>4.6	n.d.	n.d.
24 hour EC0	2.1	-	-
24 hour EC100	>4.6	-	-
48 hour EC50	3.1*	2.1	4.6
48 hour NOEC	1.0	-	-
48 hour EC0	2.1	-	-
48 hour EC100	4.6	-	-

*calculated as the geometrical mean value of the two concentrations with 0% and 100% immobility. 95% confidence value corresponds to the next lower and next higher test concentrations

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

A 48-hour EC50 value of 3.1 mg/L have been determined for the effects of the test substance on mobility of Daphnia magna.