Diammonium dihydroxybis[lactato(2-)-O1,O2]titanate(2-)

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16.07.2018 - 25.01.2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Specific details on test material used for the study:

Batch manufactured & supplied by : Dorf Ketal Chemicals India private Limited
Plot no. 2, Block-F, Sector -12N,
Adani ports & SEZ, Taluka – Mundra,
District-Kutch, Gujarat-370421

Test Item : Tyzor LA

CAS No. : 65104-06-5

Chemical name (IUPAC) : diammonium dihydroxybis[lactato(2-)-O1.O2]titanate(2-)

Titanium in product as TiO2 : 13.71 %

Physical appearance : Clear to slightly hazy.colourless to light yellow liquid

Lot No. : 334-300149/000160

Manufactured date : April 2018

Expiry date : March 2020

Recommended storage condition : Ambient (+15 to +25C)

Specific gravity : 1.215

Analytical monitoring:

not specified

Vehicle:

not specified

Details on test solutions:

6.1.3 Test Medium and Chemicals
The medium used for the test was reconstituted water, as described in the OECD test guideline 202 (OECD, 2004). The medium was prepared as per the details given in Appendix 2. The physical chemical characteristics of test medium are presented in Appendix 3.

All chemicals used in the study were of either analytical reagent (AR) or better grade reagent.

Chemical Batch No. Manufacturer
Hydroxyl ammonium chloride 1504/17101-1 Qualigens Fine Chemicals
Navi Mumbai – 400 710
India
Phenolphthalein Indicator 0000245703 Himedia laboratories Pvt. Ltd.
Mumbai – 400086, India
Sodium bicarbonate 0000230345
Magnesium sulphate. 7H2O H14A/1813/1707/31 s. d. fine-chem Limited
Mumbai-400030
India
E.D.T.A-disodium salt F10Z/1710/1705/31
Methyl orange indicator D16Z/1112/2402/74
Ammonium chloride D14A/2314/2103/31
Patton & reeders indicator F11Z/0311/1102/31
Ammonium hydroxide L11A/0711/2202/31
Calcium chloride, 2H2O D14A/1314/0804/31
Sodium hydroxide D16A/0415/2112/31
Potassium chloride QJ3P631025 Merck Specialities Private Limited
Mumbai-400018
India
Sodium carbonate QC2Q620547
Erichrome Black T AL0A601067
Sodium chloride QL4Q642763
Hydrochloric acid SHBG4416V Sigma Aldrich, 3050 Spruce street, St. Louis, USA

Test organisms (species):

Daphnia magna

Details on test organisms:

6.1.2 Test System
Daphnia magna was used as test system as it is a recommended species in the guideline.
6.1.2.1 Source
Ecotoxicology
Eurofins Advinus Limited
Bengaluru
INDIA
6.1.2.2 Maintenance of Daphnia
Daphnia were maintained in the test medium. The test medium in which the daphnia were maintained was changed at least twice a week and they were fed with the unicellular green alga, Chlorella vulgaris.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

not specified

Test temperature:

20.7 to 21.2

pH:

7.72 to 7.79

Dissolved oxygen:

6.9 to 7.4

Salinity:

not specified

Conductivity:

not specified

Nominal and measured concentrations:

6 , 12 , 25 , 50 , 100 mg/L

Details on test conditions:

6.2.1 Outline of the Method
Neonates of daphnia aged less than 24 hour old were exposed to a series of concentrations of the test item, Tyzor LA for 48 hours during range finding test and definitive test. Certain concentrations result in certain percentage of daphnia no longer being capable of swimming at 48 hours. The effects on the organisms were assessed at 24 and 48 hours interval and the EC50 value was calculated.
6.2.2 Neonates of Daphnia
One day prior to the start of the treatment, about 100 daphnia during range finding test and definitive test were incubated in approximately 2 L of test medium. The following day, the young ones hatched which were aged less than 24 hour old were collected and used in the tests.
6.2.3 Exposure to Treatment
Daphnia were exposed to the treatment under the following conditions during range finding and the definitive test.

Parameters Range finding test Definitive test
Method of exposure Static Static
Duration of the test 48 ± 1 hours 48 ± 1 hours
Test containers 250 mL glass beakers 250 mL glass beakers
No. of daphnia Ten daphnia per group divided into two replicates of five daphnia each Twenty daphnia per group divided into four replicates of five daphnia each
Loading About 15 mL of test medium to each daphnia About 30 mL of test medium to each daphnia
Light Complete darkness Complete darkness
Temperature of growth chamber (C) 20.3 to 20.5 20.7 to 21.2
Dissolved oxygen concentration of test concentration (mg/L) 7.0 to 7.4 6.9 to 7.4
pH of test concentration 7.70 to 7.80 7.72 to 7.79
Aeration The test containers were not bubble aerated The test containers were not bubble aerated
Feeding None None
6.2.4 Test Item Solubility
1 g of the test item was mixed and made up to 10 mL using test medium in a volumetric flask.
6.2.4.1 Range Finding Test
Neonates of daphnia were tested for immobility in test concentrations of Tyzor LA at 0.00001, 0.0001, 0.001, 0.01, 0.1, 1, 10 and 100 mg/L along with a negative control.
6.2.5 Definitive Test
Based on the results of the range finding test, Tyzor LA was tested at the concentrations of 6 (G2), 12 (G3), 25 (G4), 50 (G5) and 100 (G6) mg/L (factor of 2) were tested in the definitive test. Neonates of daphnia were tested for immobility in these test concentrations along with a negative control and a reference substance.
6.2.5.1 Preparation of Test Concentrations
To prepare test concentrations of 6, 12, 25, 50 and 100 mg/L, 6, 12, 25, 50 and 100 mg test item was mixed separately and volume was made up to 1 L in a volumetric flask using test medium, respectively.

1 L of test medium was used as negative control (G1).

A stock solution of reference substance (Potassium dichromate- K2Cr2O7) at 1 mg/mL was prepared by mixing 10 mg in 10 mL of test medium. For preparation of reference substance at 0.32 mg/L (G7), 0.32 mL of reference substance stock was mixed and made up to 1 L in a volumetric flask using test medium.

150 mL of negative control, test item concentrations and reference substance was dispensed into 4 separate test containers.
6.2.5.2 Stability and Analytical Verification of Test item Concentrations in Test Solutions
Stability of the test item in test medium was established for
49 h at room temperature at claimed concentrations of 0.104 and 104 mg/L using a validated analytical method.

All the test solutions along with the negative control were analyzed at the beginning of the test and at the termination of the test. For analysis, single composite sample was drawn from prepared test concentrations and duplicate samples were drawn from the negative control. Analysis was conducted using a validated analytical method in Analytical R & D Department of Eurofins Advinus Limited. The criteria for acceptance of analysis results of test concentration were 80 to 120 % of claimed concentration with ≤ 20 % RSD of analyzed concentration.
6.2.6 Observations
Following parameters were observed and recorded during both range finding test and definitive test.

pH and dissolved oxygen concentration of test concentrations at the start and end of the treatment period were recorded.

Immobility of daphnia at 24 ± 1 and 48 ± 1 hour after the start of the treatment was recorded.

The temperature of the incubator/growth chamber was recorded once daily.

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

19.373 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

9.1 Test Item Solubility
The test item formed miscible in the test medium at 100 mg/mL.
9.2 Range Finding Test
Test conditions are presented in Table 1.

Summary of daphnia immobilization presented in Table 2.

No immobility of the daphnia was observed in the negative control and at test concentrations of 0.00001, 0.0001, 0.001, 0.01, 0.1 and 1 mg/L either at 24 or at 48 hours post exposure. The immobilization of daphnia was 10 and 60 % at 24 hours and 10 and 80 % at 48 hours post exposure at the tested concentrations of 10 and 100 mg/L of Tyzor LA.
9.3 Definitive Test
Test conditions are presented in Table 3.

Summary of daphnia immobilization presented in Table 4.

Immobilization curve and dose response curves in Figures 1 to 3.

There was no immobilization of daphnia in the negative control, vehicle control and at the test concentrations of 6 mg/L either at 24 or at 48 hours exposure. The immobilization of daphnia was 10, 10, 25 and 55 % at 24 hours and 15, 30, 50 and 90% at 48 hours exposure at the tested concentrations of 12, 25, 50 and 100 mg/L of Tyzor LA, respectively.

At 0.32 mg/L reference substance, the immobilization of daphnia was 25% and 50 % observed at 24 and 48 hours exposure, respectively.

The EC50 value for Tyzor LA at 24 hours was 50.655 mg/L based on nominal concentrations with fiducial limits at 95 % being 26.2686 and 97.6789 mg/L.

The EC50 value for Tyzor LA at 48 hours was 19.373 mg/L based on nominal concentrations with fiducial limits at 95 % being 14.3068 and 26.2345 mg/L.
9.4 Analysis of Test Concentration
The results showed that the test item was stable in the test medium up to 97 hours at claimed concentrations of 0.104 and 104 mg/L (Table 5).

The active ingredient concentration analysis of all test concentrations showed that the per cent agreement with claimed concentration was 80.58 to 112.5% at start of test and 96.56 to 111.2% at end of test (48 hour) indicating that the results were within the acceptable limit (80 to 120 % of the claimed concentration with an RSD of < 20%). No measurable concentration of test item was recorded in the negative control and vehicle control samples (Table 6 to 9).

Validity criteria fulfilled:

yes

Conclusions:

The EC50 value for Tyzor LA at 48 hours was 19.373 mg/L based on nominal concentrations with fiducial limits at 95 % being 14.3068 and 26.2345 mg/L

Executive summary:

The acute immobilization effect of the test item Tyzor LA was studied onDaphnia magna. TheDaphnia magnaof less than 24 hours old were exposed to concentrations of 6, 12, 25, 50 and 100 mg/L of Tyzor LA along with a negative control and reference substance in reconstituted water at 20 ± 1^oC. The number of daphniaimmobilized was recorded at 24 and 48 hours post exposure.

 

There was no immobilization of daphnia in the negative control, vehicle control and at the test concentrations of 6 mg/L either at 24 or at 48 hours exposure. The immobilization of daphniawas 10, 10, 25 and 55 % at 24 hours and 15, 30, 50 and 90% at 48 hours exposure at the tested concentrations of 12, 25, 50 and 100 mg/L ofTyzor LA, respectively.

 

At 0.32 mg/Lreference substance, the immobilization of daphniawas 25% and 50 % observed at 24 and 48 hours exposure, respectively.

 

The results showed that the test item was stable in the test medium up to 97 hours at claimed concentrations of 0.104 and 104 mg/L. The active ingredient concentration analysis of all test concentrations showed that the per cent agreement with claimed concentration was 80.58 to 112.5% at start of test and 96.56 to 111.2% at end of test (48 hour) indicating that the results were within the acceptable limit (80 to 120 % of the claimed concentration with an RSD of<20%).

 

The EC₅₀value for Tyzor LA at 24 hours was 50.655 mg/L based on nominal concentrations with fiducial limits at 95 % being 26.2686 and 97.6789 mg/L.

 

The EC₅₀value for Tyzor LA at 48 hours was 19.373 mg/L based on nominal concentrations with fiducial limits at 95 % being 14.3068 and 26.2345 mg/L.

 

Description of key information

The EC₅₀value for Tyzor LA at 48 hours was 19.373 mg/L based on nominal concentrations with fiducial limits at 95 % being 14.3068 and 26.2345 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

19.373 mg/L