D-Glucitol, 1-deoxy-1-(methylamino)-, N-C8-10 acyl derivs.

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-10-10 to 2013-10-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study performed under GLP, no deficiencies

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-10(even numbered) acyl] derivs.

EC Number:

940-284-1

Cas Number:

1591782-62-5

Molecular formula:

C15H31NO6 C17H35NO6

IUPAC Name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-10(even numbered) acyl] derivs.

Constituent 2

Reference substance name:

Glucamide 810

IUPAC Name:

Glucamide 810

Details on test material:

- Physical state: Colourless powder
- Sum formula: C15H31NO6 (C8-Glucamide) C17H35NO6 (C10-Glucamide)
- Molecular weight:
C8-Glucamide: 321.42 g/Mol
C10-Glucamide 349.47 g/Mol
- Storage condition of test material: Room temperature, protected from light, in original container.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The limit concentration of 100 mg/L is based on the results of preliminary range finding tests.
- Sampling method:
Analytical evaluation of the limit concentration of C8/C10-Glucamide AC93/13 was carried out via LC-MS/MS from freshly prepared test media at start of the exposure (0 h) and from corresponding 96 h aged test media at the end of the exposure.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Directly weighing. The test media was mixed with an ultraturrax (1 min, 17000 rpm).

- Eluate: Dilution water
- Differential loading: Limit concentration of 100 mg/L
- Controls: 7 fish in dilution water (without test item) were tested under the same test conditions as the test replicates.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: All fish used in the test were gained at DR.NOACK-LABORATORIEN from a single brood stock. (supplier: Umweltbundesamt, Schichauweg 58, D-12307 Berlin)
- Length at study initiation (length definition, mean, range and SD): Average body length: 2.74 cm
- Weight at study initiation (mean and range, SD): Average body weight: 0.28 g
- Method of breeding:Holding was performed at the test facility at 23 +/- 2 °C and diffuse light (0.1 - 10 µmol photons • m-2 • s-1, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value. No disease treatments were administered throughout holding and testing. Food was provided 3 times per week. The amount of food was 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started.
- Feeding during test: No feeding during test



ACCLIMATION
- Acclimation period: Zebrafish with at least 12 days of acclimatisation and mortality < 5 % within these days before the study starts were used in the test.
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started. Food: Sera Vipan; SERA GMBH, D-52518 Heinsberg
- Feeding frequency: 3 times per week.
- Health during acclimation (any mortality observed): No mortality observed, no disease treatments were administered throughout holding and testing.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

Total Hardness at day 0: 43 mg/L

Test temperature:

Please refer to "Any other information on materials and methods"

pH:

Please refer to "Any other information on materials and methods"

Dissolved oxygen:

Please refer to "Any other information on materials and methods"

Salinity:

Not measured, freshwater

Nominal and measured concentrations:

Please refer to "Any other information on materials and methods incl. tables"

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria
- Type (delete if not applicable): open, loosely covered by glass tops
- Material, size, headspace, fill volume: glass, 10 L test volume per vessel
- Aeration: Gentle aeration
- Renewal rate of test solution (frequency/flow rate): None, static design
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: less than 1 g fish per L test solution


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water of local origin was used for holding and testing. The water was filtered on activated charcoal and aerated for at least 24 h to remove possible chlorine.
Nominal water parameters:
Total hardness: 10 - 250 mg CaCO3/L
pH-value: 6.0 - 8.5
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH-value, temperature and oxygen saturation were measured in all vessels at the beginning of the test and every 24 h. Total hardness of the water was determined at the beginning out of the test control. During the test the water temperature was recorded continuously with a data logger (once per hour).


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: natural
- Light intensity: 0.1 - 10 µmol photons x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.). Observations were made after 24, 48, 72 and 96 h.


TEST CONCENTRATIONS
- Range finding study:
Cumulative Mortality [%] in the Preliminary Test
(n = 5)
Nominal Test Item Concentration
[mg/L] Cumulative mortality [%] after different exposure periods [hours]
24 48 72 96
100 0 0 0 0
Control 0 0 0 0

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: highest test item concentration with 0% mortality after 96h

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: Not observed
- Mortality of control: 0%
- Other adverse effects control: Not observed
- Abnormal responses: Not observed
- Any observations (e.g. precipitation): The test solution was clearly dissolved throughout exposure.

Results with reference substance (positive control):

No reference substance tested

Reported statistics and error estimates:

The concentrations leading to 0 and 100 % mortality (LC0 and LC100) after 96 h were determined directly from the test results. LC10,20,50-values had not to be calculated in a limit test.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Observations in the Definitive Test

Nominal test item concentration [mg/L]	Obser-vations *	Number of fish effected at observation time [hours]
		24	48	72	96
100	(1)	7/7	7/7	7/7	7/7
Control	(1)	7/7	7/7	7/7	7/7

Cumulative Mortality [%] in the Definitive Test

Nominal test item concentration [mg/L]	Cumulative mortality at observation time [hours]
	24	48	72	96
100	0	0	0	0
Control	0	0	0	0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In this study, C8/C10-Glucamide AC93/13 was found to cause no lethal effects to Zebrafish after 96 hours at a nominal test concentration of 100 mg/L (highest tested concentration).
The LC50 after 96 hours is > 100 mg/L. All effect levels are given based on the nominal concentrations of the test item C8/C10-Glucamide AC93/13.

Executive summary:

The acute toxicity of the test item C8/C10-Glucamide AC93/13(batch no.:AC93/13) to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from 2013-10-10 to 2013-10-14 at Dr.U.Noack-Laboratorien, 31157 Sarstedt, Germany.

 

A static test was performed with nominal test item concentration of 100 mg/L. Duration of the test was 96 hours. 7 test organisms were exposed to each concentration and the control. Water quality parameters pH-value, temperature and oxygen-saturation, measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits.

 

The concentrations of the test item C8/C10-Glucamide AC93/13 were analytically verified by LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (96 h) of the limit concentration and the control.
The measured concentration of the test item at the start of the exposure (0 h) was 94 % of the fraction C8-Glucamid and 96 % of the fraction C10-Glucamid of the nominal value. The measured concentrations of the test item at the end of the exposure (96 h) was 105 % of the fraction C8-Glucamid and 103 % of the fraction C10-Glucamid of the nominal value.

    

LC-Values (0 – 96 hours)

     Based on nominal test item concentrations [mg/L].

Test duration [hours]	C8/C10-Glucamide AC93/13[mg/L]
	LC10	LC20	LC50
24	> 100	> 100	> 100
48	> 100	> 100	> 100
72	> 100	> 100	> 100
96	> 100	> 100	> 100
LC0= Highest test item concentration with 0 % mortality after 96 hours	100
LC100= Lowest test item concentration with 100 % mortality after 96 hours	> 100