Sodium p-[(4,6-dichloro-1,3,5-triazin-2-yl)amino]benzenesulphonate

Long-term toxicity to aquatic invertebrates

Description of key information

At the limit concentration of 143 mg/L of the test item Active Substance of Granofin Easy F-90 liq in Aqueous Solution, no biologically or statistically significant effects were determined. The No Observed Effect Concentration (NOEC) and the Lowest Observed Effect Concentration (LOEC) were assessed as follows: 
NOEC	:	   143 mg/L
LOEC	:	> 143 mg/L
All provided effect values are based on the nominal concentration of the test item Active Substance of Granofin Easy F-90 liq in Aqueous Solution.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

143 mg/L

Additional information

TheDaphnia magnaReproduction Test (Semi-Static, 21 d) of Active Substance of Granofin Easy F-90 liq in Aqueous Solution was conducted as a limit test with the concentration of 143 mg/L of the test item, which corresponds to 100 mg/L of the main component, according to OECD 211 (2008) from 2012-05-30 to 2012-06-21, with the definitive exposure phase from2012-05-30 to 2012-06-20atDr.U.Noack-Laboratorien,Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany.
The selected test organism wasDaphnia magnaSTRAUS (Clone 5). Ten daphnids individually held were used for the limit concentration and the control. At the test start the daphnids were 2 to 24 h old. The test method was semi-static with a renewal of the test solutions three times per week. The aim of the Daphnia Reproduction Test over 21 days was to assess effects on the reproduction capacity and other test item-related effects, including occurrence of aborted eggs and stillborn juveniles, time of production of first brood, adult mortality, intrinsic rate of natural increase, dry body weight and length of the parent daphnids.
The tested limit concentration of the test item Active Substance of Granofin Easy F-90 liq in Aqueous Solution and the control were analytically verified by HPLC-DAD of samples on days 0, 7, 16 (fresh media, 0 hours) and on days 2, 9, 19 (old media, 48 and 72 hours). Details of the analytical method are presented in part 11.

The measured concentrations of the test item Active Substance of Granofin Easy F-90 liq in Aqueous Solution were in the range of 94 to 101 % of the nominal values in samples of the fresh media (0 hours), and 87 to 93 % of the nominal values in the old media (48 and 72 hours) of the limit concentration and the control. The results given are based on the nominal concentration of 143 mg/L of the test item Active Substance of Granofin Easy F-90 liq in Aqueous Solution.

·  The averagevalues ofjuveniles per parental daphnidwere 123 for the control and 121 for the limit concentration of 143 mg/L after 21 days. The reproductive output of daphnids in the limit concentration was comparable to the reproductive output of daphnids in the control(t-test,p = 0.05).

·  The coefficient of variation of thenumber of living offspringproduced per parent was 10 % for the control and 7 % for thelimit concentration

·  Theintrinsic rates of natural increase (IR)of the surviving parent daphnids accounting for generation time and number of offspring were calculated as a measure for population growth and maintenance. The IR of thelimit concentrationwas comparabletothe control group (t-test,p = 0.05).

·  Noadult mortalityor immobilization of parental daphnids was observedin the control andthelimit concentration.

·  Nodead juveniles and aborted eggswere observed in the control and thelimit concentration.

Five broods were observed during the test period for all parental daphnids of the limit concentration and the control. Thefirst broodwas released from all parent daphnids in all treatments between day 8 and 9.

·  The mean drybody weight and total body lengthsof all parental daphnids of the test and the control groups were determined at the end of the study.
Themeandry body weightof the parental daphnids was 1.06 mg per daphnid in the control and 0.77 mg per daphnid in the limit concentration.
Themean bodylengthof the parental daphnids was 5.20 mm per daphnid in the control and 5.00 mm per daphnid in the limit concentration.

·  Nomalesandephippia(winter eggs) were observed in the respective control or test groups.

A summary of all test item related effects is given inTable 1. No biologically or statistically significant effect could be determined in the limit concentration of 143 mg/Lcompared to the control.

Table1:   Summary of all Test Item Related Effects

(Based on the nominal concentration of the test itemof 143 mg/L)

Active Substance of Granofin Easy F-90 liq in Aqueous Solution
Effects	Control	Nominal Concentration 143 mg/L	Significance
Mean Number of Juveniles per Producing Parent (Reproduction Rate±SD)	123 ± 12	121 ± 8	no^1)
Coefficient of Variation of theMean Number of Juveniles per Producing Parent	10	7	no
Intrinsic Rate of Natural Increase (Mean±SD)	0.48 ± 0.03	0.49 ± 0.01	no^1)
Appearance of First Brood (Mean Day±SD)	8.1 ± 0.3	8.0 ± 0.0	—
Number of Broods	5	5	—
Stillborn Juveniles and Aborted Eggs	0	0	—
Adult Mortality after 21 Days [%]	0	0	no
Parent Daphnids: Mean Dry Weight [mg]	1.06	0.77	—
Parent Daphnids: Mean Body Length [mm]±SD	5.20 ± 0.26	5.00 ± 0.17	no
NOEC	143 mg/L
LOEC	> 143 mg/L

Thewater quality parameters(pH-value, dissolved oxygen, water hardness and temperature) were within the acceptable limits.

·         To assure thevalidity of the test systemand the test conditions at the test facility, an acute immobilization test according to DIN 38412 L 11 is carried out withpotassium dichromateas reference item once per month. This reference test is required by the test guideline OECD 211 (2008).
The EC₅₀of the reference item of1.74 mg/L (95 % confidence limits: 1.49 - 2.00 mg/L)after 24 h is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according toAQS 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.