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EC number: 701-162-1
CAS number: -
of Juveniles in the Control and Test Groups after 21 Days
Mean no. of juveniles
Number of living juveniles in replicate no.
MV ± SD
[%] of the Adult Daphnids after 7, 14 and 21 Days of Exposure
test item concentration
Adult mortality [%]
Number of Dead (Stillborn + Aborted Eggs) to Total Number of Juveniles
Number of juveniles
Appearance of Living Juveniles in the Individual Groups
Day of first appearance of living juveniles
in replicate no.
Body Length of the Parental Daphnids
Total length of the parent animals
magna Reproduction Test (semi-static, 21 d) of the test item (Pentapropylensuccinimido)-capronic
acid (batch number:
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,
1, 31157Sarstedt, Germany.
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
Nominal concentrations of
acid were selected as follows: 1.25 - 2.50 - 5.00 - 10.0 -
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.
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 % .
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.
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.
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.
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.
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.
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.
or ephippia (winter eggs) were
observed in the control or in the test groups.
water quality parameters(i.e. pH-value, dissolved oxygen
concentration, total water hardness and temperature) were determined to
be within the acceptable limits.
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
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.
of all test item related effects and the assessed endpoints based on the
nominal test item concentrations is given in Table 1.
Related Effects (NOEC, LOEC, EC10, EC50 and EC100)
on the nominal concentrations of the test item)
Nominal test item concentrations [mg/L]
Mean Number of Living Juveniles per
Producing Parent (Reproduction Rate ± SD)
87 ± 5
85 ± 6
Coefficient of Variation of the Mean Number of Juveniles per Producing Parent [%]
Mean Number of Broods
Appearance of First Brood [Mean Day]
Mean Intrinsic Rates of Natural Increase
Percentage of Dead Juveniles Related to the Total Number of Juveniles [%]
Parental Daphnids: Mean Dry Weight [mg]
Parental Daphnids: Mean Body Length [mm]
Adult Mortality after 21 Days [%]
1.70 mg/L (95 % confidence limits: 1.25 – 5.00 mg/L)
2.49 mg/L (95 % confidence limits: 1.25 – 5.00 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
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