6-[C12-18-alkyl-(branched, unsaturated)-2,5-dioxopyrrolidin-1-yl]hexanoic acid

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-02-19 to 2013-03-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study, 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 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

6-[3-(C12-C18)-alkenyl(branched, unsaturated)-2,5-dioxopyrrolidin-1-yl]hexanoic acid

EC Number:

701-162-1

Molecular formula:

C22H37NO4 - C28H49NO4

IUPAC Name:

6-[3-(C12-C18)-alkenyl(branched, unsaturated)-2,5-dioxopyrrolidin-1-yl]hexanoic acid

Details on test material:

- Name of test material (as cited in study report): (Pentapropylensuccinimido)-capronic acid
- Physical state: Liquid, brown viscous
- Analytical purity: 95.0 % (w/w) (100 % minus by-products)
- Purity test date: 2011-11-15
- Appearance: Brown, viscous, liquid
- Lot/batch No.: ESD0009272
- Expiration date of the lot/batch: 2014-04-26
- Stability under test conditions: Not specified
- Storage condition of test material: Room temperature, protected from light, in original container

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1.25 - 2.50 - 5.00 - 10.0 - 20.0 mg/L
- Sampling method:
All concentration levels of the test item and the control were analysed by LC-MS at the start of the exposure intervals on days 0, 9, 14 (0 h) and at the end of the exposure intervals on days 2, 12, 16 (48 or 72 h). For the analyses of the old media, separate replicates without daphnids and algae were prepared and stored under test conditions until sampling.

- Sample storage conditions before analysis: All original samples were stored at room temperature until sample preparation. Prepared samples were stored at room temperature until analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution of 20.0 mg/L of the test item was prepared one day prior to each water renewal in demineralised water adjusted with 1 M NaOH to a pH of 11 ± 0.2.
40.0 mg of the test item were weighed out and mixed with 1900 mL of the pH-adjusted demineralised water (pH 11 ± 0.2).
The stock solution was stirred with approximately 1100 rpm for 24 hours at room temperature.
After the stirring period, the mineral components as specified under 'Test medium' were added to yield the highest test concentration.
- Eluate: Dilution water
- Differential loading: 1.25 - 2.50 - 5.00 - 10.0 - 20.0 mg/L
- Controls: 10 replicates of dilution water without test item

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUS
- Strain/clone: Clone 5
- Justification for species other than prescribed by test guideline: Daphnia magna STRAUS is recommended in the guideline
- Source: Own breeding (Origin: Institut fuer Wasser- Boden- und Lufthygiene)
- Age of parental stock (mean and range, SD): > 14 days
- Feeding during test
- Food type: Mix of Pseudokirchneriella subcapitata and Desmodesmus subspicatus
- Amount: 0.2 mg C/daphnia per day
- Frequency: renewals 3 times per week


ACCLIMATION
- Acclimation period: 2 h in dilution water
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: During acclimation the daphnids were not fed
- Feeding frequency: None
- Health during acclimation (any mortality observed): Healthy

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: The parent animals were removed from the culture medium and the juveniles collected over a sieve and flushed into fresh medium

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

Not observed

Test conditions

Hardness:

Total Hardness [mg/L] as CaCO3
Nominal
test item
concentration
[mg/L] I F I F I F
Day 0 Day 2 Day 9 Day 12 Day 14 Day 16
2013-02-20 2013-02-22 2013-03-01 2013-03-04 2013-03-06 2013-03-08
20.0 169 146 Not determined due to 100 % adult mortality
2.50 Not determined 169 185 142 141
Control 169 158 164 160 164 157

Test temperature:

Temperature [°C]
Nominal
test item
concentration
[mg/L] I F I F I F
Day 0 Day 2 Day 9 Day 12 Day 14 Day 16
2013-02-20 2013-02-22 2013-03-01 2013-03-04 2013-03-06 2013-03-08
20.0 21.0 21.0 Not determined due to 100 % adult mortality.
2.50 Not determined 21.0 20.1 19.8 19.8
Control 20.2 21.0 19.3 20.1 19.4 19.9

pH:

pH-Values
Nominal
test item
concentration
[mg/L] I F I F I F
Day 0 Day 2 Day 9 Day 12 Day 14 Day 16
2013-02-20 2013-02-22 2013-03-01 2013-03-04 2013-03-06 2013-03-08
20.0 7.12 7.72 Not determined due to 100 % adult mortality
2.50 Not determined 7.82 8.66 7.83 7.71
Control 8.34 7.96 8.02 8.21 8.11 7.86

Dissolved oxygen:

Dissolved Oxygen Concentration [mg/L]
Nominal
test item
concentration
[mg/L] I F I F I F
Day 0 Day 2 Day 9 Day 12 Day 14 Day 16
2013-02-20 2013-02-22 2013-03-01 2013-03-04 2013-03-06 2013-03-08
20.0 8.60 8.33 Not determined due to 100 % adult mortality
2.50 Not determined 8.75 10.76 8.86 8.23
Control 8.96 8.76 9.01 9.12 9.19 8.52

Salinity:

Not measured, freshwater

Nominal and measured concentrations:

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

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: 100 mL glass beaker, filled with 50 mL test solution
- Aeration: No
- Renewal rate of test solution (frequency): 3 x per week
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light : 8 h dark per day
- Light intensity: Max. 20 µE*m-2*s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Adult mortality: daily
- Number of juveniles: daily
- Stillborn juveniles and aborted eggs: daily
- Appearance of first brood: between days 8 and 9
- Intrinsic rate of natural increase: test end
- Growth (total length and dry weight): test end


RANGE-FINDING STUDY

Immobilisation Rates in the Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal
Concentration
of the Test Item
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 100 100 100 100 100 100
10.0 10 0 5 30 10 20
1.00 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1.25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals:
The test item induced biologically significant adult mortality at the concentration levels of 2.50 mg/L (90 %) and 5.00 to 20.0 mg/L (100 %) during the test period of 21 days. In the concentration level of 1.25 mg/L and in the control, all parental daphnids survived until the end of the test after 21 days.
The EC10 for the adult mortality was calculated to be 1.93 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L). The EC50 for the adult mortality was calculated to be 2.19 mg/L (95% confidence limits: 1.25 – 5.00 mg/L). The EC100 for the adult mortality was 5.00 mg/L.
- No. of offspring produced per day per female:
The average number of living juveniles per surviving parental daphnid at the end of the test after 21 days was 87 in the control group. The reproductive output was statistically significant reduced in comparison to the control at the concentration level of 2.50 mg/L (One Way Analysis of Variance, DUNNETT’S method, p = 0.05). At the concentration level of 1.25 mg/L, the reproductive output was comparable to the reproductive output at the control. Due to 100 % adult mortality, no offspring was observed at the concentration levels of 5.00 to 20.0 mg/L.
Based on the significant reduction of the reproductive output and the biologically significant adult mortality of 90 % at the concentration level of 2.50 mg/L, the No Observed Effect Concentration (NOEC) after 21 days was assessed at 1.25 mg/L and the Lowest Observed Effect Concentration (LOEC) was assessed at 2.50 mg/L. The EC10-value for the reduction of the reproductive output was calculated to be 1.70 mg/L (95 confidence limits: < 1.25 – 5.00 mg/L). The EC50-value for the reduction of the reproductive output was calculated to be 2.49 mg/L (95 % confidence limits: < 1.25 – 5.00 mg/L).
- Body length and weight of parent animals:
The mean values of the body length of the surviving parental daphnids at the tested concentration levels of 1.25 and 2.50 mg/L were 5.23 and 4.50 mm per daphnid. A mean body length of 5.40 mm per daphnid was determined for the control group.
The mean dry body weight was determined to be 0.72 and 0.40 mg per daphnid at the concentration levels of 1.25 and 2.50 mg/L and 0.99 mg per daphnid at the control
- Type and number of morphological / behavioural abnormalities:
please refer to the respective table
- Time to first brood release or time to hatch:
The first brood was released on days 8 and 9 by all parental daphnids of the control and the concentration levels of 1.25 and 2.50 mg/L. At the tested concentration level of 1.25 mg/L and in the control, 4 broods were produced by the surviving parental daphnids. The surviving daphnid at the concentration level of 2.50 mg/L released only 3 broods.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: EC50 (24 h) = 1.23 (CI 1.14 - 1.37) mg/L

Reported statistics and error estimates:

The NOEC and the LOEC based on the reduction of the reproductive output and the intrinsic rates of natural increase were deduced directly from the results. Significant deviations were determined in comparison to the control using statistical standard procedures as normality test, equal variance test and analysis of variance (One Way Analysis of Variance, DUNNETT’S method).
Prior to running the One Way Analysis of Variance, a normality test and an equal variance test were performed. P-values of the normality and equal variance tests should be 0.05. The actual P-values were > 0.05. The a-value (acceptable probability of incorrectly concluding that there is a difference) was a=0.05.
The coefficients of variation around the mean number of living offspring produced per parent in the control and the test groups were evaluated.

Any other information on results incl. tables

Number of Juveniles in the Control and Test Groups after 21 Days

Nominal test item conc. [mg/L]												No. of parents prod. juv.	Mean no. of juveniles per parent producing juveniles			Comparison versus control
	Number of living juveniles in replicate no.										Total no.				CV
																Red.	Stat.
	1	2	3	4	5	6	7	8	9	10	å	N	MV ± SD		[%]	[%]
20.0	--	--	--	--	--	--	--	--	--	--	n.a.	0	n.a.		n.a.	n.a.	n.a.
10.0	--	--	--	--	--	--	--	--	--	--	n.a.	0	n.a.		n.a.	n.a.	n.a.
5.00	--	--	--	--	--	--	--	--	--	--	n.a.	0	n.a.		n.a.	n.a.	n.a.
2.50	--	--	--	--	--	--	--	--	--	43	43	1	43 ±	—	—	51	yes
1.25	89	79	80	92	80	88	79	92	94	80	853	10	85 ±	6	7	2	no
Control	95	86	89	86	90	81	84	82	92	84	869	10	87 ±	5	5	—	—

Mortality [%] of the Adult Daphnids after 7, 14 and 21 Days of Exposure

                 (n = 10)

Nominal test item concentration [mg/L]	Adult mortality [%]
	7 days	14 days	21 days
20.0	100^1)	100^1)	100^1)
10.0	100^1)	100^1)	100^1)
5.00	100^1)	100^1)	100^1)
2.50	70^1)	90^1)	90^1)
1.25	0	0	0
Control	0	0	0

Relative Number of Dead (Stillborn + Aborted Eggs) to Total Number of Juveniles

Nominal test item concentration [mg/L]	Number of juveniles			Percentage of dead juveniles^#) [%]
	Dead	Alive	Total
20.0	n.a.	n.a.	n.a.	n.a.
10.0	n.a.	n.a.	n.a.	n.a.
5.00	n.a.	n.a.	n.a.	n.a.
2.50	7	43	50	14
1.25	4	853	857	1
Control	0	869	869	0

First Appearance of Living Juveniles in the Individual Groups

Nominal test item concentration [mg/L]	Day of first appearance of living juveniles										First	Mean
	in replicate no.										appearance	number
	1	2	3	4	5	6	7	8	9	10	mean day	of broods
20.0	--	--	--	--	--	--	--	--	--	--	n.a.	n.a.
10.0	--	--	--	--	--	--	--	--	--	--	n.a.	n.a.
5.00	--	--	--	--	--	--	--	--	--	--	n.a.	n.a.
2.50	--	--	--	--	--	--	--	--	--	9	9.0	3
1.25	9	9	8	8	8	9	9	9	9	9	8.7	4
Control	9	8	9	8	8	8	9	9	9	9	8.6	4

Total Body Length of the Parental Daphnids

Nominal test item concentration [mg/L]	Total length of the parent animals [mm]										MV	N	Dry weight [mg]
	Replicate no.
	1	2	3	4	5	6	7	8	9	10	[mm]		S	MV
20.0	--	--	--	--	--	--	--	--	--	--	n.a.	0	n.a.	n.a.
10.0	--	--	--	--	--	--	--	--	--	--	n.a.	0	n.a.	n.a.
5.00	--	--	--	--	--	--	--	--	--	--	n.a.	0	n.a.	n.a.
2.50	--	--	--	--	--	--	--	--	--	4.50	4.50	1	0.40	0.40
1.25	5.25	4.75	5.00	5.25	5.50	5.25	5.50	5.50	5.25	5.00	5.23	10	7.2	0.72
Control	5.50	5.00	5.50	5.50	4.75	5.25	5.50	5.50	5.75	5.75	5.40	10	9.9	0.99

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the statistically significant reduction of the reproductive output, the statistically significant intrinsic rate and the biologically significant adult mortality of 90 % at the concentration level of 2.50 mg/L, the No Observed Effect Concentration (NOEC) after 21 days was assessed at 1.25 mg/L and the Lowest Observed Effect Concentration (LOEC) at 2.50 mg/L.
The EC10-value for the reduction of the reproductive output was calculated to be 1.70 mg/L (95 confidence limits: < 1.25 – 5.00 mg/L). The EC50-value for the reduction of the reproductive output was calculated to be 2.49 mg/L (95 % confidence limits: < 1.25 – 5.00 mg/L).
The EC10 for the adult mortality was calculated to be 1.93 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L). The EC50 for the adult mortality was calculated to be 2.19 mg/L (95% confidence limits: 1.25 – 5.00 mg/L. The EC100 for the adult mortality was 5.00 mg/L.

Endpoints assessed
(Pentapropylensuccinimido)-capronic acid
Nominaltest item concentration
[mg/L]
EC10 Reproduction 1.70 mg/L (95 % confidence limits: < 1.25 – 5.00 mg/L)
EC50 Reproduction 2.49 mg/L (95 % confidence limits: < 1.25 – 5.00 mg/L)
NOECReproduction 1.25 mg/L
LOECReproduction 2.50 mg/L
EC10 Adult Mortality 1.93 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
EC50 Adult Mortality 2.19 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
EC100 Adult Mortality 5.00 mg/L

Executive summary:

The Daphnia magna Reproduction Test (semi-static, 21 d) of the test item (Pentapropylensuccinimido)-capronic acid (batch number: ESD0009272) was conducted according to OECD 211 (2008) from 2013-02-19 to 2013-03-14, with the definitive exposure phase from 2013-02-20 to 2013-03-13, at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str. 1, 31157Sarstedt, Germany.

Test species was Daphnia magna STRAUS (Clone 5). Ten daphnids, individually held, were used per concentration level and control. At the test start, the daphnids were 2 to 24 hours old. The study was carried out under semi-static conditions with renewal of the test solutions three times per week. Aim of the Daphnia magnaReproduction Test over 21 days was to assess effects on the reproduction capacity and other test item-related effects or parameters such as time of production of first brood, adult mortality, intrinsic rate of natural increase, occurrence of aborted eggs and stillborn juveniles and body length of the parental daphnids.

Nominal concentrations of (Pentapropylensuccinimido)-capronic acid were selected as follows: 1.25 - 2.50 - 5.00 - 10.0 - 20.0 mg/L.

The concentrations of the test item (Pentapropylensuccinimido)-capronic acid were analytically verified by LC-MS of samples taken at the start of the exposure intervals on days 0, 9, 14 (0 h) and at the end of the exposure intervals on days 2, 12, 16 (48 or 72 h) of all concentration levels and the control.

The measured concentrations of the test item at the start of the exposure intervals were in the range of 75 to 104 % of the nominal values. At the end of the exposure intervals (48 or 72 h), the measured concentrations of the test item were in the range of 70 to 100 % of the nominal values.
The effect levels are based on the nominal concentrations, because recoveries in the range of ± 20 % of the nominal values were typically observed. Two samples with recoveries of 70 and 75 % were regarded to be outliers and have therefore no influence on the test results.

·   The average number of living juveniles per surviving parental daphnid at the end of the test after 21 days was 87 in the control group. The reproductive output was statistically significant reduced in comparison to the control at the concentration level of 2.50 mg/L (One Way Analysis of Variance, Dunnett’smethod, p = 0.05). At the concentration level of 1.25 mg/L, the reproductive output was comparable to the reproductive output at the control. Due to 100 % adult mortality, no offspring was observed at the concentration levels of 5.00 to 20.0 mg/L.
The EC₁₀-value for the reduction of the reproductive output was calculated to be 1.70 mg/L (95 % confidence limits:  1.25 – 5.00 mg/L). The EC₅₀-value for the reduction of the reproductive output was calculated to be 2.49 mg/L (95 % confidence limits:  < 1.25 – 5.00 mg/L).

·   Based on the significant reduction of the reproductive output and the biologically significant adult mortality of 90 % at the concentration level of 2.50 mg/L, the No Observed Effect Concentration (NOEC)after 21 days was assessed at 1.25 mg/L and the Lowest Observed Effect Concentration (LOEC) was assessed at 2.50 mg/L.

·   The coefficient of variation of the number of living offspring produced per parent was 5 % at the control and 7 % at the concentration level of 1.25 mg/L. For the concentration levels of 2.50 to 20.0 mg/L, no coefficient of variation could be calculated because of the adult mortality of ≥ 90 % .

·   The test item induced biologically significant adult mortality at the concentration levels of 2.50 mg/L (90 %) and 5.00 to 20.0 mg/L (100 %) during the test period of 21 days. In the concentration level of 1.25 mg/L and in the control, all parental daphnids survived until the end of the test after 21 days. The EC₁₀ for the adult mortality was calculated to be 1.93 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L). The EC₅₀ for the adult mortality was calculated to be 2.19 mg/L (95% confidence limits: 1.25 – 5.00 mg/L. The EC₁₀₀ for the adult mortality was 5.00 mg/L.

·   No stillborn juveniles or aborted eggs were observed in the control group during the exposure period of 21 days. A small number of aborted eggs of 4 at the concentration level of 1.25 mg/L and 7 at the concentration level of 2.50 mg/L were observed.

·   The intrinsic rates of natural increase (IR) of the surviving parental daphnids accounting for generation time and number of offspring were used for calculation of population growth and maintenance. The intrinsic rates of natural increase determined for the surviving daphnids at the concentration levels of 1.25 to 2.50 mg/L were compared to the IR at the control (One Way Analysis of Variance, Dunnett’s method, p = 0.05). There was a statistical significance at the concentration level of 2.50 mg/L. The IR at the concentration level of 1.25 mg/L was comparable to the IR at the control.

·   The first brood was released on days 8 and 9 by all parental daphnids of the control and the concentration levels of 1.25 and 2.50 mg/L. At the tested concentration level of 1.25 mg/L and in the control, 4 broods were produced by the surviving parental daphnids. The surviving daphnid at the concentration level of 2.50 mg/L released only 3 broods.

·   The mean values of the body length of the surviving parental daphnids at the tested concentration levels of 1.25 and 2.50 mg/L were 5.23 and 4.50 mm per daphnid. A mean body length of 5.40 mm per daphnid was determined for the control group.

     The mean dry body weight was determined to be 0.72 and 0.40 mg per daphnid at the concentration levels of 1.25 and 2.50 mg/L and 0.99 mg per daphnid at the control.

·       No males or ephippia (winter eggs) were observed in the control or in the test groups.

 

·   The water quality parameters(i.e. pH-value, dissolved oxygen concentration, total water hardness and temperature) were determined to be within the acceptable limits.

In order to prove the validity of the test systemand test conditions at the test facility, an acute immobilisation test according to DIN 38412 L 11 is monthly carried out with potassium dichromate as reference item.
The EC₅₀ of the reference item was 1.23 mg/L (95 % confidence limits: 1.14 - 1.37 mg/L) after 24 hours and therefore within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according to AQS P 9/2 (05/1996) 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. For details see part 4.6.

A summary of all test item related effects and the assessed endpoints based on the nominal test item concentrations is given in Table 1.

Table 1:           Test Item Related Effects (NOEC, LOEC, EC₁₀, EC₅₀ and EC₁₀₀)

                         (based on the nominal concentrations of the test item)

Effects	(Pentapropylensuccinimido)-capronic acid Nominal test item concentrations [mg/L]
	Control	1.25	2.50	5.00	10.0	20.0
Mean Number of Living Juveniles per Producing Parent (Reproduction Rate ± SD)	87 ± 5	85 ± 6	43^1)	---	---	---
Coefficient of Variation of the Mean Number of Juveniles per Producing Parent [%]	5	7	n.a.	---	---	---
Mean Number of Broods	4.00	4.00	3.00	---	---	---
Appearance of First Brood [Mean Day]	8.6	8.7	9.0	---	---	---
Mean Intrinsic Rates of Natural Increase	0.45	0.45	0.38^1)	---	---	---
Percentage of Dead Juveniles Related to the Total Number of Juveniles [%]	0	1	14	---	---	---
Parental Daphnids: Mean Dry Weight [mg]	0.99	0.72	0.40	---	---	---
Parental Daphnids: Mean Body Length [mm]	5.40	5.23	4.50	---	---	---
Adult Mortality after 21 Days [%]	0	0	90^2)	100^2)	100^2)	100^2)
EC10 Reproduction	1.70 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
EC50 Reproduction	2.49 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
NOECReproduction	1.25 mg/L
LOECReproduction	2.50 mg/L
EC10 Adult Mortality	1.93 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
EC50 Adult Mortality	2.19 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
EC100 Adult Mortality	5.00 mg/L