Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental result of of read across substance

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

WoE report is based on two short term toxicity study of algae for the test chemical :1. and 2. To evalute algal growth inhibition test of test material

GLP compliance:

not specified

Specific details on test material used for the study:

IUPAC name: Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate Molecular weight: 515.7375Molecular formula: C33H45N3O2Smiles: CCN(CC)c1ccc(C{+}(c2ccc(N(CC)CC)cc2)(c2ccc(N(CC)CC)cc2).O{-}C(C)=O)cc1Inchl: 1S/C31H42N3.C2H4O2/c1-7-32(8-2)28-19-13-25(14-20-28)31(26-15-21-29(22-16-26)33(9-3)10-4)27-17-23-30(24-18-27)34(11-5)12-6;1-2(3)4/h13-24H,7-12H2,1-6H3;1H3,(H,3,4)/q+1;/p-1Physical state: Solid and liquidForm: Organic

Analytical monitoring:

not specified

Vehicle:

not specified

Details on test solutions:

1.The stock solution (10 mg/L) was prepared by dissolving yellow powder in OECD growth medium. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture.2.The test item was prepared by adding 12 mg of test item in 240 ml of BBM to get the final concentration of 50mg/L. This stock solution was kept for stirring for 30 minutes to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment.

Test organisms (species):

other: 1.Desmodesmus subspicatus , 2.Chlorella vulgaris

Details on test organisms:

1.TEST ORGANISM- Common name: - Strain: 86.81 SAG- Source (laboratory, culture collection): Institute of botany of the ASCR- Initial biomass concentration: 5x10(3) cells /ml - Method of cultivation: No data available2.The fresh water green alga Chlorella vulgaris, was used as the test organism. Sterile, unicellular, liquid cultures of algae.The culture was examined under the microscope to confirm that it was unicellular, healthy and not contaminated. The medium to be used for the growth of algae was Bold’s Basal Medium (BBM). It is a medium composed of macronutrients, micronutrients, alkaline EDTA solution and Iron solution. Stock solution of each of these was prepared separately and then a complete medium was prepared and sterilized. De-ionized water was used to prepare the BBM.

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Test temperature:

1) 23±2°C2) 24 °C ±2°C.

pH:

Sample at concentration 0.64 mg/L: pH= 7.9 changed to pH= 7.8 during the testControl: pH= 8.0 changed to pH= 7.4 during the test2)6.7 - 7.4

Nominal and measured concentrations:

1) 0, 0.02, 0.04, 0.08, 0.16, 0.32 and 0.64 mg/L.2) 0.5 mg/l, 1.25mg/l,3.125mg/l, 7.813mg/l, 19.531mg/l and 48.828mg/l

Details on test conditions:

1.TEST SYSTEM- Test vessel: 50 ml Glass vessel- Type (delete if not applicable): closed (with air permeable stopper)- Sample volume: 15 ml- Initial cells density: 5x10(3) cells/ml- No. of vessels per concentration (replicates): 3GROWTH MEDIUM- Standard medium used: yesOTHER TEST CONDITIONS- Adjustment of pH: No- Photoperiod: Continuous- Light intensity and quality: 6000-8000 lxEFFECT PARAMETERS MEASURED (with observation intervals if applicable) :- Determination of cell concentrations: microscope with counting chamber Cyrus I or electronic particle counter.- Other: ErC50 was calculated using non-linear regression by the software Prism 4.02.Light QualityWhite Fluoroscent LightLight Intensity1500LuxPhotoperiod16 Hour Light Period : 8 Hour Dark PeriodRPM Speed120 Revolutions per minuteWater Deionized WaerTest vessel size100mlTest solution volume60mlDilution waterBold’s Basal MediumTest concentration Six test concentration were: Replicates per concentrationThree replicates for ControlTwo replicates for each test concentration

Reference substance (positive control):

yes

Remarks:

K2Cr2O7

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

0.13 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95% CL: 0.11 - 0.15 mg/L for 1 study

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

0.61 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: For 2 nd study

Details on results:

2.The microscopic observations were also noted in each of the experimental flasks. All the cells appeared healthy, round and green throughout the test duration in the control and in the experimental flask also no significant changes were observed.

Results with reference substance (positive control):

Results with reference substance valid- EC50: 0.75 mg/L

Validity criteria fulfilled:

not specified

Conclusions:

1. The median effective concentration (EC50) for the test substance, in Desmodesmus subspicatus was determined to be 0.13 mg/L on the basis of effects on growth rate in a 72 hour study.2. After 72 hours of exposure to test item to various nominal test concentrations, EC50 was found to be 0.610mg/l graphically and through probit analysis.Thus, comparing this value with the criteria of CLP regulation Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as toxic for short term for aquatic algae and cyanobacteria as and classified aq aquatic acute 1 as per the CLP criteria.

Executive summary:

Data available for the structurally similar read across chemicals has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate.The studies are as mentioned below:

1.Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the test substance according to OECD Guideline 201.The stock solution (10 mg/L) was prepared by dissolving yellow powder in OECD growth medium. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. The test substance was dissolved in DMSO/ OECD growth medium and tested at the concentrations 0, 3, 6, 12, 24 and 50 mg/L.Determination of cell concentrations was performed by microscope with counting chamber Cyrus I or electronic particle counter.Effects on the growth rate of the organism were studied. The median effective concentration (EC50) for the test substance, test substance, in Desmodesmus subspicatus was determined to be 0.13 mg/L. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as Aquatic acute category 1 as per the CLP criteria.

2.The effect of test item was studied on the growth of fresh water green algaChlorella vulgaris.The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 0.5mg/l, 1.25mg/l, 3.125mg/l, 7.813mg/l, 19.531mg/l and 48.828mg/l. The test concentrations were prepared using stock solution of the test item using mineral media. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be 0.610 mg/L.Thus, based on the EC50 value, test substancecan be considered astoxic to aquatic organisms and thus can be classified as aquatic acute category 1 as per the CLP criteria.

Thus, based on the above summarised studies, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be concluded that effect concetration value is in the range of 0.13 - 0.61 mg/ L. Thus, comparing this value with the criteria of CLP regulation Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as toxic for short term for aquatic algae and cyanobacteria as and classified aq aquatic acute 1 as per the CLP criteria.

Description of key information

1. The median effective concentration (EC50) for the test substance, in Desmodesmus subspicatus was determined to be 0.13 mg/L on the basis of effects on growth rate in a 72 hour study.

2. After 72 hours of exposure to test item to various nominal test concentrations, EC50 was found to be 0.610mg/l graphically and through probit analysis.

Thus, comparing this value with the criteria of CLP regulation Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as toxic for short term for aquatic algae and cyanobacteria as and classified aq aquatic acute 1 as per the CLP criteria.

Key value for chemical safety assessment

EC50 for freshwater algae:

0.13 mg/L

Additional information

Data available for the structurally similar read across chemicals has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate.The studies are as mentioned below:

 

In the key study from the experimental report 2016 freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the test substance according to OECD Guideline 201.The stock solution (10 mg/L) was prepared by dissolving yellow powder in OECD growth medium. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. The test substance was dissolved in DMSO/ OECD growth medium and tested at the concentrations 0, 3, 6, 12, 24 and 50 mg/L. Determination of cell concentrations was performed by microscope with counting chamber Cyrus I or electronic particle counter. Effects on the growth rate of the organism were studied. The median effective concentration (EC50) for the test substance, test substance, in Desmodesmus subspicatus was determined to be 0.13 mg/L. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as Aquatic acute category 1 as per the CLP criteria.

 

The first study was supported by the second experimental study for read across chemical. The effect of test item was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 0.5mg/l, 1.25mg/l, 3.125mg/l, 7.813mg/l, 19.531mg/l and 48.828mg/l. The test concentrations were prepared using stock solution of the test item using mineral media. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be 0.610 mg/L. Thus, based on the EC50 value, test substance can be considered as toxic to aquatic organisms and thus can be classified as aquatic acute category 1 as per the CLP criteria.

 

Thus, based on the above summarized studies, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate it can be concluded that effect concentration value is in the range of 0.13 - 0.61 mg/L. Thus, comparing this value with the criteria of CLP regulation reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as toxic for short term for aquatic algae and cyanobacteria as and classified aquatic acute 1 as per the CLP criteria.