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

Endpoint summary

Additional information

SHORT-TERM TOXICITY TO FISH

The acute toxicity of the test item Active substance of Granofin Easy F-90 liq in aqueous solution (batch no.:ITA0056831)to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from 2012-03-12 to 2012-03-30 with the definite exposure phase from 2012-03-26 to 2012-03-30, atDr.U.Noack-Laboratorien, 31157 Sarstedt, Germany. A static test with one limit concentration of nominal 143 mg test item/L, corresponding to 100 mg main component/L, was performed.

The duration of the test was 96 h. Seven test organisms were exposed to the limit concentration and the control. Water quality parameters pH-value, temperature and oxygen-saturation measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits.

The determination of the test item Active substance of Granofin Easy F-90 liq in aqueous solution was carried out via HPLC-DAD from the limit concentration and the control at test start and test end. The recovery rates of Active substance of Granofin Easy F-90 liq in aqueous solution at start and end of the test were 103 % and 92 % of the nominal value, respectively. For details of the analytical results please refer to part 8. All effect levels are given based on the nominal concentration of the test item Active substance of Granofin Easy F-90 liq in aqueous solution (Table 1).

LC-Values with 95 % Confidence Intervals

                        based on nominal test item concentration [mg/L]

Test duration [h]	LC50	p = 95 %
24	> 143	n.a.
48	> 143	n.a.
72	> 143	n.a.
96	> 143	n.a.
LC100=	> 143
Lowest test item concentration
with 100 % mortality after 96 h
LC0=	143
Highest test item concentration
with 0 % mortality after 96 h

LONG-TERM TOXICITY TO FISH

Waiving according to "column 2" in Annex IX of REGULATION (EC) No 1907/2006 (CSA does not indicate need for further investigations. There are no acute toxicity effects on aquatic organisms). Nevertheless a chronic study with daphnia magna is available.

SHORT-TERM TOXICITY TO AQUATIC INVERTEBRATES

The test substance was tested for its acute toxicity to daphnids. The test article was found to be non-Toxic to daphnids (EC50: > 100 mg/L).

LONG-TERM TOXICITY TO AQUATIC INVERTEBRATES

TheDaphnia magnaReproduction Test (Semi-Static, 21 d) of Active Substance of Granofin Easy F-90 liq in Aqueous Solution (batch number ITA0056831) 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. TOXICITY TO AQUATIC ALGAE

The toxicity ofActive substanceof Granofin Easy F-90 liq inaqueous solution (batch number ITA0056831)to the unicellular freshwater green algaDesmodesmus subspicatuswas determined according to the principles of OECD 201atDr.U.Noack-Laboratorienin 31157 Sarstedt, Germanyfrom to , with the definitive exposure phase from 2012-03-27 to 2012-03-30.The aim of the study was to assess the effects on growth rate and yield over a period of 72 hours.

The study was conducted under static conditions with an initial cell density of 4910 cells/mL.Based on a preliminary range finding test the limit concentration of 143 mg test item /L, corresponding to 100 mg/L main component,was tested.Sixreplicates were tested for the limit concentration and the control.Environmental conditionswere determined to be within the acceptable limits.

The limit concentration and the control were analysed by HPLC-DAD after (fresh media) and 72 h (old media).

The measured concentration of the test item in the fresh media (0 h) was 101 %. The test item concentration in the old media (72 h) was 83 %.

All effect values are based on the nominal concentration of the main component as well as the test item.

NOEC, LOEC, EC - values of Active substanceof Granofin Easy F-90 liq inaqueous solution (0-72 hours)

                  based on nominal concentration of the main component and test item [mg/L]

	Inhibition of Growth Rate
	Main Component [mg/L]	Test Item                   [mg/L]
NOEC	100	143
LOEC	> 100	> 143
ErC10	> 100	> 143
ErC20	> 100	> 143
ErC50	> 100	> 143
	Inhibition of Yield
NOEC	100	143
LOEC	> 100	> 143
EyC10	> 100	> 143
EyC20	> 100	> 143
EyC50	> 100	> 143

TOXICITY TO MICROORGANISMS

A Respiration Inhibition Test with activated sludge according to OECD Guideline No. 209 was carried out with the test item Active Substance of Granofin Easy F-90 liq in aqueous solution (batch number:ITA0056831)from 2012-05-23 to 2012-05-24, with the definitive experimental phase on 2012-05-24, at theDr.U.Noack-Laboratorien, D-31157Sarstedt.Test system was activated sludge of the municipal treatment plant of D-31137 Hildesheim. The test was carried out under static conditions with the substance concentrations 100 - 180 - 320 - 580 - 1000 mg a.i./L, corresponding to 145 - 261 - 464 - 840 - 1449 mg/L. The respiration rates of the control, reference and test item replicates were measured after a contact time of three hours, and the inhibitory effects of the test and reference item were determined in comparison to the control respiration rates. The mean inhibition of respiration for the test item replicates ranged from -10 % to 41 %.

In order to check the activity of the test system and the test conditions a reference test was carried out with copper (II) sulphate pentahydrate as reference item and the reference toxicity was determined. The EC₅₀-value for the reference item was 88.7 mg/L.

EC-Values with Confidence Interval of Active Substanceof Granofin Easy F-90 liq in aqueous solution

	EC-values [mg a.i./L]	Confidence interval P = 95 [%] [mg a.i./L]
NOEC*	320	–
EC20	554	516 - 593
EC50	> 1000	–
EC80	> 1000	–

The NOEC of the test itemActive Substanceof Granofin Easy F-90 liq in Aqueous Solution is320 mg a.i./L.

The test item is not toxic up to the concentration of320mg a.i./L to activated sludge of a municipal sewage treatment plant. The EC₅₀of the test item is> 1000 mg a.i./L.

In a second Respiration Inhibition Test with activated sludge according to OECD 209 it was shown, that the test item is not toxic up to the highest tested concentration (IC50 > 100 mg/L). This supports the results from the kew study.