1-(3-chlorophenyl)-4-(3-chloropropyl)piperazinium chloride

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 6-7, 2012

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1-(3-chlorophenyl)-4-(3-chloropropyl)piperazinium chloride

EC Number:

258-038-0

EC Name:

1-(3-chlorophenyl)-4-(3-chloropropyl)piperazinium chloride

Cas Number:

52605-52-4

Molecular formula:

C13H18Cl2N2.ClH

IUPAC Name:

1-(3-chlorophenyl)-4-(3-chloropropyl)piperazin-1-ium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Test item identification : 1-(3-chloroprophyl)-4-(3-chlorophenyl)-piperazine hydrochloride
Active ingredient content : 97.4 % w/w
Batch number : 0900002043
Manufacturing date : June 07, 2011
Expiry date : December 07, 2012
Stability : Stable under the storage conditions.

Test solutions

Vehicle:

yes

Details on test solutions:

The main test included six test item concentrations in a geometric series, namely 1.9, 4.3, 9.4, 20.7, 45.5 and 100.0 mg/L into reconstituted water .

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

The study was performed on the freshwater crustacean Daphnia magna Straus. A strain of this species was originally supplied by the Italian Health Department (Istituto Superiore di Sanità) in September 2004. Since that time, the clone has been bred in the laboratories of the Test Facility in reconstituted water of the quality identical to the water quality used in the tests (regarding pH, main ions, and total hardness) and under temperature and light
conditions identical to those of the tests (see below). It was daily fed with a fixed amount of the laboratory cultured green algae Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) and with a suspension of the yeast Saccharomyces cerevisiae.
At the test start the organisms used in the test were less than 24 hours old and were not first brood progeny.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

Water Hardness : 140 - 250 mg/L as CaCO3

Test temperature:

The incubation temperature was continuously monitored during the course of the study by the mean of a data logger. It was in the range
18.1 – 20.6 °C with a mean value of 19.3 °C and a standard deviation of 0.8 °C. The temperature range provided by OECD guideline is 20 ± 2 C,
this range was corrected according to the data logger precision (0.48°C), therefore the range to be considered for the test was 18.5 – 21.5 °C .The
measured values were slightly lower than the corrected range and the temperature variation was higher than 1.0 °C; however these deviations
did not affected the test results since occurred for short periods and it had not any effects on the daphnia mobility in negative control.

pH:

The pH values of the test media after preparation were in the range 6.26 – 7.22 for test item solutions and 7.14 for the negative control. The pH did not vary by more than 0.45 unit during the test period, as required in the mentioned guideline

Dissolved oxygen:

The dissolved oxygen concentrations in the test media and the negative control were always higher than 7.0 mg/L according to OECD Guideline N 202 that provides a minimum oxygen concentration of 3.0 mg/L.

Details on test conditions:

Light: a 16 hours light and 8 hours darkness daily photoperiod. The measured values were in the range 854 – 864 Lux, according to provided range
(700-1000 Lux).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate solution

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

IC50

Effect conc.:

9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 6.7-12.2 %

Any other information on results incl. tables

In the negative control 0% of immobilization was observed and no daphnid was found trapped on the test water surface.

These values comply with the validity criterion of the test, that provide a maximum immobilization and/or a maximum number of daphnids trapped into water surface of 10% in the negative control medium at the end of the test. The obtained experimental results allowed to determine the IC0, IC50 and the IC100 values at 24, and 48 h.

The IC0 and IC100 values at each observation time were directly extracted from the Raw Data.

The results, based on the nominal concentrations of test item 1-(3-chloroprophyl)-4-(3 - chlorophenyl)-piperazine hydrochloride, were as follows:

Time (h)	IC0 (mg/L)	IC100 (mg/L)
24	1.9	> 100.0
48	< 1.9	100.0

At each observation time, the IC50 was calculated by Trimmed Spearman-Karber method.

The statistical analysis was chosen by CETIS software as the most appropriate.

The IC50 for each observation time with its 95% confidence limits (LCL, Lower Confidence

Level and UCL, Upper Confidence Level) determined using the nominal concentrations of test item as input data were as follows:

Time	IC50 (mg/L)	95% LCL (mg/L)	95% UCL (mg/L)
24 h	75.1	47.7	118.4
48 h	9.0	6.7	12.2

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The IC50 with its 95% confidence limits (LCL, Lower Confidence Level and UCL, Upper Confidence Level) at 24 and 48 hours, calculated in terms of
nominal test item concentrations, were as follows:
IC50 (mg/L) 24 h= 75.1 (47.7-118.4)
IC50 (mg/L) 48 h= 9.0 (6.7-12.2)

Executive summary:

The acute toxicity of the test item 1-(3-chloroprophyl)-4-(3-chlorophenyl)-piperazine hydrochloride to Daphnia magna was determined in a 48-hour full static test according to the OECD Guideline for Testing of Chemicals, No. 202 (2004) and to Council Regulation EC 440/2008 (C.2).

For this purpose, juvenile daphnids (< 24 hours old at initiation of the test) were exposed to an aqueous test medium containing the test item at six different concentrations, namely 1.9, 4.3, 9.4, 20.7, 45.5 and 100.0 mg/L.

The exposed organisms were checked for immobilization 24 and 48 hours after test initiation.

The pH of the test media after preparation was in the range 6.26 – 7.22 for test item solutions and 7.14 for negative control.

The incubation temperature was in the range 18.1 – 20.6 °C and the light intensity during the 48 hours of test period was in the range 854 - 864 Lux.

At the end of test period, in the negative control 0 % of immobilization was observed and no daphnid was found to be trapped on the test water surface. These values comply with the validity criterion of the test, that provide a maximum immobilization and/or a maximum number of daphnids trapped into water surface of 10% in the negative control medium at the end of the test.

The obtained experimental results allowed to assess the IC0, IC100 and the IC50 values with its 95% confidence limits (LCL, Lower Confidence Level and UCL, Upper Confidence Level) at 24 and 48 h.

The results, based on the nominal concentrations of test item, were as reported in the following table:

Time (h) IC0 (mg/L) IC100 (mg/L) IC50 with 95% Confidence range (mg/L)

24 1.9 > 100.0 75.1 (47.7 – 118.4)

48 < 1.9 100.0 9.0 (6.7 – 12.2)

The IC50 values were determined from the Raw Data by the statistical analysis in comparison

with the negative control (Trimmed Spearman-Karber method) while IC0 and IC100 values at

each observation time were directly extracted from the Raw Data.