Fatty acids, C18-unsatd., compds. with diisopropanolamine N-C12-18-alkyl derivs.

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:

2014-12-17 to 2014-12-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, no deficiencies

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.

Cas Number:

2156594-85-1

Molecular formula:

Not applicable

IUPAC Name:

Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.

Test material form:

other: viscous liquid

Details on test material:

- Name of test material (as cited in study report): Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.
- Physical state: Brown , slightly turbid, viscous liquid
Storage condition of test material: Room temperature, protected from light, in original container

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

All concentration levels and the control were analytically verified by LC-MS/MS at the start of both exposure intervals (0 and 24 h) and at the end of both exposure intervals (24 and 48 h).
At the start of the exposure intervals (0 and 24 h), sampling was carried out after preparation of the test concentrations.
At the end of the exposure intervals (24 and 48 h), samples were taken directly from the test vessels. Adsorption was determined as specified below. The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).

Recoveries of the test item should be within ± 20 % of the nominal or initially measured concentrations.
However, the effect levels given are based on the nominal concentrations of the test item Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.

Determination of losses due The sorption of the test item on glass was quantified at the
to adsorption onto glassware concentration levels of 1.00 – 2.00 – 4.00 mg/L (double determinations) at the end of both exposure intervals (24 and 48 h). Two additional replicates per concentration level without daphnids were prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied and an appropriate amount of a suitable solvent was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration. Before this procedure, the test vessels were carefully rinsed with demineralised water to remove the remaining test media.

Schedule of Specific Test Item Analysis
Sampling Date Samples intended to be measured:
[h]

0 and 24 h
(fresh media) aqueous phase
without daphnids:
all test media and control
24 and 48 h
(old media) aqueous phase
with daphnids:
all test media and control
aqueous phase and glass
surface without daphnids:
1.00 – 2.00 – 4.00 mg/L

Test solutions

Vehicle:

no

Details on test solutions:

The stock solution (8.00 mg/L of the test item were weighed out) was freshly prepared with dilution water prior to each exposure interval (0 and 24 h)
The stock solution was treated with ultrasound for 15 minutes at 40 °C and stirred thereafter with approximately 1100 rpm for 30 minutes at room temperature.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Clone 5
- Source:
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder: DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
- Age at study initiation (mean and range, SD): 2 to 24 h old daphnids from a healthy stock were used for the study
- Method of breeding:
Culture: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 +- 2°C, in an incubator, 16 h illumination; light intensity
of max. 20 µE x m-2 x s-1
Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
Cultur feeding: At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and
Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: At least 2 h in dilution water.
- Acclimation conditions (same as test or not): Same as test conditions.
- Health during acclimation (any mortality observed): No mortality observed.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

Dilution Water Total Hardness
Dated: [mg CaCO3/L]
0 h: 255
2014-12-17
24 h: 262
2014-12-18

Test temperature:

18 - 22°C, constant within ± 1°C

pH:

pH at the Start of the Exposure Intervals (0 and 24 h)
(measured in one additional replicate per concentration level and control)
Nominal
test item
concentration
[mg/L] 0 h 24 h
pH-value pH-value

8.00 7.60 7.60
4.00 7.62 7.62
2.00 7.66 7.67
1.00 7.68 7.69
0.500 7.70 7.69
0.250 7.73 7.71
Control 7.61 7.64



pH at the End of the 1st Exposure Interval (24 h)
(measured in all replicates)

Nominal
test item
concentration
[mg/L] pH-values
Replicates
1 2 3 4
8.00 7.48 7.47 7.47 7.45
4.00 7.48 7.46 7.45 7.47
2.00 7.47 7.47 7.46 7.46
1.00 7.48 7.47 7.47 7.48
0.500 7.45 7.44 7.49 7.47
0.250 7.49 7.47 7.48 7.45
Control 7.60 7.53 7.49 7.48

pH at the End of the 2nd Exposure Interval (48h)
(measured in all replicates)

Nominal
test item
concentration
[mg/L] pH-values
Replicates
1 2 3 4
8.00 7.55 7.57 7.57 7.56
4.00 7.58 7.56 7.56 7.56
2.00 7.58 7.57 7.56 7.59
1.00 7.59 7.59 7.57 7.58
0.500 7.62 7.61 7.61 7.56
0.250 7.66 7.64 7.63 7.63
Control 7.67 7.69 7.68 7.67

Dissolved oxygen:

O2-concentration at the Start of the Exposure Intervals (0 and 24 h)
(measured in one additional replicate per concentration level and control)

Nominal
test item
concentration
[mg/L] 0 h 24 h
Dissolved Dissolved
O2-concentration [mg/L] O2-concentration [mg/L]

8.00 8.14 7.76
4.00 8.67 8.40
2.00 8.92 8.72
1.00 9.02 8.88
0.500 9.07 8.95
0.250 9.08 8.95
Control 9.17 8.90




O2-concentration at the End of the 1st Exposure (24 h)
(measured in all replicates)

Nominal
test item
concentration
[mg/L] Dissolved O2 -concentration [mg/L]
Replicates
1 2 3 4
8.00 8.32 8.50 8.38 8.40
4.00 8.54 8.38 8.31 8.35
2.00 8.65 8.33 8.43 8.49
1.00 8.49 8.51 8.53 8.50
0.500 8.65 8.63 8.55 8.62
0.250 8.68 8.66 8.68 8.57
Control 8.67 8.57 8.50 8.52

O2-concentration at the End of the 2nd Exposure (48 h)
(measured in all replicates)

Nominal
test item
concentration
[mg/L] Dissolved O2 -concentration [mg/L]
Replicates
1 2 3 4
8.00 8.48 8.64 8.68 8.25
4.00 8.33 8.52 8.30 8.51
2.00 8.60 8.60 8.56 8.62
1.00 8.62 8.45 8.58 8.68
0.500 8.71 8.49 8.49 8.65
0.250 8.72 8.73 8.74 8.79
Control 8.73 8.56 8.68 8.72

Nominal and measured concentrations:

6 test item concentrations in a geometric series with a separation factor of 2, prepared by diluting the stock solution of 8.00 mg/L with dilution water, were tested as follows:
0.250 - 0.500 - 1.00 - 2.00 - 4.00 - 8.00 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 mL capacity, loosely covered with watch glasses
- Test volume: 20 mL
- Aeration: None
- 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: ISO Test water, according to OECD 202, Annex 3
Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2 x 2 H2O 294
MgSO4 x 7 H2O 123

- Water Quality Parameters of the Dilution Water prior to the Start of the Exposure Intervals (0 and 24 h):
Dilution Water
Dated: pH-Value Dissolved O2-Concentration [mg/L] Temperature [°C] Conductivity [µS/cm] Total Hardness [mg CaCO3/L]
0 h:2014-12-17 7.61 9.17 19.5 639 255
24 h:2014-12-18 7.64 8.90 20.9 643 262


- Culture medium different from test medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement:interval start (0 and 24 h) and interval end (24 and 48h)

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 µE x m-2 x s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Immobilisation, 24 h interval

TEST CONCENTRATIONS
- Test concentrations: 0.250 - 0.500 -1.00 - 2.00 - 4.00 - 8.00 mg/L (spacing factor 2 )

- Range finding study:
A non GLP preliminary range finding test under semi-static conditions over a period of 48 hours was conducted at the test facility with three nominal test item concentrations of 0.100, 1.00 and 10.0 mg/L. In the range finding test, two replicates per treatment group and control, each with ten daphnids, were tested. The biologically results are presented in the table below. The results of the analytical monitoring during the range finding test are presented in the tables below

Immobilisation Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)

Nominal test item concentration
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
10.0 90 100 95 100 100 100
1.00 0 0 0 0 0 0
0.100 0 0 0 0 0 0
Control 0 0 0 0 0 0

MV = mean value


During the range finding test, the test solutions were clear. The exposure concentrations were monitored at the start (0 and 24 h) and at the end (24 and 48 h) of the exposure intervals in test vessel with daphnids and test vessels without daphnids.

Measured Concentrations of the Component Test Item during the non GLP Preliminary Range Finding Test

Sampling date 2014-11-19 2014-11-20 2014-11-20 2014-11-21
0 h 24 h 24 h 48 h
Start of the End of the Start of the End of the
exposure interval exposure interval exposure interval exposure interval
Nominal
test item
concentration
[mg/L] Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.
Meas. Meas. Meas. Meas.
conc. conc. conc. conc.
[mg/L] % [mg/L] % [mg/L] % [mg/L] %
10.0 10.2 102 7.06 71 8.02 80 7.94 79
1.00 1.11 111 0.869 87 1.04 104 1.06 106
0.100 0.118 118 0.0827 83 0.104 104 0.0792 79
Control < 0.0100 < 0.0100 < 0.0100 < 0.0100
Meas. conc. = measured concentration, mean value of 2 injections, dilution factor taken into account
% = percent of the nominal concentration of the test item


The sorption of the test item on glass was quantified at the end of both exposure intervals (24 and 48 h). For this purpose, an additional replicate per concentration level without daphnids was prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied and an appropriate amount of a suitable solvent was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration. Before this procedure, the test vessels were carefully rinsed with demineralised water to remove the remaining test media.

Measured Concentrations of the Test Item in Aqueous Phase and at Glass Walls of Test Vessels without Daphnids during the non GLP Preliminary Range Finding Test

Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-,
N-C12-18-(even numbered) alkyl derivs.
Nominal End of the 1st exposure interval End of 2nd exposure interval
test item (24 h) (48 h)
concentration Aqueous Phase Adsorbed Aqueous Phase Adsorbed
[mg/L] 20 mL to Glass 20 mL to Glass
Meas. Meas. Meas. Meas
conc. conc.* conc. conc.*
[mg/L] % [mg/L] % [mg/L] % [mg/L] %
10.0 7.78 78 1.02 10 7.95 80 0.823 8
1.00 0.905 91 0.0406 4 0.885 89 0.0314 3
0.100 0.0828 83 < 0.0100 0.0606 61 < 0.0100
Meas. conc. = measured concentration, mean value of 2 injections, dilution factor taken into account
% = percent of the nominal concentration of the test item

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

2.63 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 2.36 - 2.91

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

4.46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 4.26 - 4.70

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

See below

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC50-value with 95 % confidence limits (Cl) was calculated by sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was:
EC50: 2.05 mg/L (Cl 1.90 - 2.18 mg/L)
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of 0.6 - 2.4 mg/L of quality criteria according to AQS P 9/2 (02/2000) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Any other information on results incl. tables

Biological Data

The percentage immobility, determined in all test and control groups after 24 and 48 h under semi-static conditions, is given in the table below.

 

Immobilisation Rates after 24 and 48 h of Exposure in the Definitive Test

(n = 20, divided into 4 replicates with 5 daphnids each)

Nominal test item concentrations   [mg/L]	IMMOBILISATION [%]
	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
8.00	20	0	20	0	10	100	80	80	100	90
4.00	0	0	0	0	0	40	40	40	40	40
2.00	0	0	0	0	0	0	0	0	0	0
1.00	0	0	0	0	0	0	0	0	0	0
0.500	0	0	0	0	0	0	0	0	0	0
0.250	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

MV = mean value

 

All test concentrations were visually clear throughout the exposure period.

Measured Exposure Concentrations during the Definitive Test

The concentrations of the test itemwereanalytically verified via LC-MS/MS at the start of the exposure intervals (0 and 24 h) and at the end of the exposure intervals (24 and 48 h) in all concentration levels and the control.

The measured concentrations of the test item at the start of the exposure intervals (0 and 24 h) were in the range of 78 to 112 % of the nominal values. At the end of the exposure intervals (24 and 48 h), the measured concentrations of the test item were in the range of 72 to 90 % of the nominal values.The analytical results arepresented in the table below.

Adsorption to the glassware was found negligible. However, the measured concentrations were successfully maintained for the duration of the test at the biologically relevant test item concentrations.

Measured Concentrations of theTest ItemFatty acids, C18-unsatd., compd.with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.during the Definitive Test

Sampling date	2014-12-17 Start of the exposure interval 0 h		2014-12-18 End of the exposure interval 24 h		2014-12-18 Start of the exposure interval 24 h		2014-12-19 End of the exposure interval 48 h
Start of analysis	2014-12-17		2014-12-18		2014-12-18		2014-12-19
Nominal test item concentration [mg/L]	Fattyacids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.
	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
8.00	6.59^1)	82	6.82	85	8.53	107	7.09	89
4.00	3.41^1)	85	3.24	81	4.48	112	3.55	89
2.00	1.55^1)	78	1.43^2)	72	2.09	105	1.69	85
1.00	0.790^1)	79	0.722^2)	72	1.03^3)	103	0.896	90
0.500	0.408^1)	82	0.402	80	0.551	110	0.398	80
0.250	0.207^1)	83	0.184^2)	74	0.261	104	0.184^2)	74
Control	< LOQ		< LOQ		< LOQ		< LOQ

Meas. conc.= measured concentration of the test item, mean value of 2 injections, dilution factor taken into account

%                  = percent of the nominal concentration of the test item

LOQ             = limit of quantification (20.0 µg/L of the test item)

¹⁾                   = reanalysis on 2014-12-19, mean value of 2 replicates with 2 injections each, dilution factor taken into account

²⁾                   = reanalysis on 2014-12-19, mean value of 3 replicates with 2 injections each, dilution factor taken into account

³⁾                   = reinjected on 2014-12-18

 

Adsorption to Glassduring the Definitive Test

 

Adsorption to the glass walls of the test vessels was determined exemplarily at the concentration levels of 1.00 – 2.00 – 4.00 mg/L (double determinations) in separate replicates without daphnids at the end of both exposure intervals (24 and 48 h).

Two additional replicates per concentration level without daphnids were prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied, rinsed carefully with demineralised water and an appropriate amount of a suitable solvent was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration.

Adsorption to the glass walls was in the range of 3 – 7 % of the nominal test item concentrations. For details, see the table below

Fatty acids, C18-unsatd., compd.with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.
	Aqueous Phase 20 mL		Adsorbed to Glass		Aqueous Phase 20 mL		Adsorbed to Glass
	End of the 1^stexposure interval (24 hours)		after exposure of 24 hours		End of the 2^ndexposure interval (24 hours)		after exposure of 24 hours
	Meas. conc. [mg/L]	%	Meas. conc.* [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc.* [mg/L]	%
4.00	3.29	82	0.154	4	3.73	93	0.267	7
2.00	1.69	85	0.0800	4	1.88	94	0.102	5
1.00	0.802	80	0.0273	3	0.860	86	0.0353	4

Meas. conc.= measured concentration of the test item, mean value of 2 injections, dilution factor taken into account

%                  = percent of the nominal concentration of the test item

LOQ             = limit of quantification (20.0 µg/L of the test item)

*             = mean value of 2 replicates with 2 injections each

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item Fatty acids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs., the 48 h-EC50 for Daphnia magna was 4.46 mg/L (95 % confidence limits: 4.26 – 4.70 mg/L).
The NOEC after 48 h was 2.00 mg/L. The LOEC after 48 h was 4.00 mg/L.

Executive summary:

In the acute immobilisation test withDaphnia magna (STRAUS), the effects of the test item Fatty acids, C18-unsatd., compd.with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.(batch number:DEG4273438) were determined according to OECD 202 (2004) from2014-12-17 to 2014-12-19 at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str.1, 31157 Sarstedt, Germany.

The study was conducted under semi-static conditions over a period of 48 h with6concentrations of the test itemFatty acids, C18-unsatd., compd.with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.in the range of 0.250 to 8.00 mg/Lin a geometric series with a separation factor of 2.

The test item is a mixture of salt and ester of primary fatty amine propoxylate and fatty acid and forms an emulsion in water up to a maximum of 181 mg/L (20 °C). The test item is a brown, slightly turbid and viscous liquid.All test concentrations were visually clear throughout the exposure period.

Twenty daphnids were exposed to each concentration leveland the control.

The concentrations of the test itemwereanalytically verified via LC-MS/MS at the start of the exposure intervals (0 and 24 h) and at the end of the exposure intervals (24 and 48 h) in all concentration levels and the control.

The measured concentrations of the test item at the start of the exposure intervals (0 and 24 h) were in the range of 78 to 112 % of the nominal values. At the end of the exposure intervals (24 and 48 h), the measured concentrations of the test item were in the range of 72 to 90 % of the nominal values..

Adsorption to the glass walls of the test vessels was determined exemplarily at the concentration levels of 1.00 – 2.00 – 4.00 mg/L (double determinations) in separate replicates without daphnids at the end of both exposure intervals (24 and 48 h).
Adsorption to the glass walls was in the range of 3 – 7 % of the nominal test item concentrations. .

Adsorption to the glassware was found negligible. However, the measured concentrations were successfully maintained for the duration of the test at the biologically relevant test item concentrations. The effect levels given are based on the nominal concentrations of the test itemFatty acids, C18-unsatd., compd.with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 and 24 h) and at the end of the exposure intervals (24 and 48 h), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.  

Based onthe nominal concentrationsof the test itemFatty acids, C18-unsatd., compd.with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.,

the 48 h-EC₅₀forDaphnia magnawas 4.46 mg/L (95 % confidence limits: 4.26 – 4.70 mg/L).

The NOEC after 48 h was 2.00 mg/L.The LOEC after 48 h was 4.00 mg/L.

 

EC₁₀-, EC₅₀- (with Confidence Interval), EC₁₀₀-Values, NOEC and LOEC

(based on the nominal concentrations of the test item)

Fattyacids, C18-unsatd., compd. with 2-propanol, 1,1’-iminobis-, N-C12-18-(even numbered) alkyl derivs.
Effect levels	Test duration [h]	Nominal test item concentrations [mg/L]
EC10 (with 95 % confidence limits)	24	8.00	(Cl: Not applicable)
	48	2.63	(Cl: 2.36 – 2.91)
EC50 (with 95 % confidence limits)	24	> 8.00	(Cl: Not applicable)
	48	4.46	(Cl: 4.26 – 4.70)
EC100	24	> 8.00
	48	> 8.00
NOEC	48	2.00
LOEC	48	4.00