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EC number: 701-162-1 | CAS number: -
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
Mortality [%] of the Adult Daphnids after 7, 14 and 21 Days of Exposure
(n = 10)
test item concentration
Adult mortality [%]
Relative Number of Dead (Stillborn + Aborted Eggs) to Total Number of Juveniles
Number of juveniles
First Appearance of Living Juveniles in the Individual Groups
Day of first appearance of living juveniles
in replicate no.
Total Body Length of the Parental Daphnids
Total length of the parent animals
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)
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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