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Toxicity to aquatic algae and cyanobacteria

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Reference
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Justification for type of information:
Data is from the exeprimental report followed standard gudeline.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Principles of method if other than guideline:
Determination of the effect of Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex(CAS No12225-21-7) on thegrowth of freshwater green alga Chlorella vulgaris (OECD Guideline No.201).
GLP compliance:
not specified
Specific details on test material used for the study:
- Name of test material (IUPAC name): aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate)- Common name: FD&C Yellow No. 5 Aluminum Lake- Molecular formula: C48H33AlN12O27S6- Molecular weight: 495.4038 g/mol-InChl:1S/C16H12N4O9S2.Al/c21-15-13(18-17-9-1-5-11(6-2-9)30(24,25)26)14(16(22)23)19-20(15)10-3-7-12(8-4-10)31(27,28)29;/h1-8,13H,(H,22,23)(H,24,25,26)(H,27,28,29);/b18-17+;- Substance type: Organic- Physical state: solid
Analytical monitoring:
no
Details on test solutions:
The test solution was prepared in aseptic condition. The test item Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring/sonication 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 concentrations chosen were set up to the water solubility limit. The remaining test solutions were prepared by dilution from the above stock solution. To have abetter growth and visibility of cells, the initial of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment.
Test organisms (species):
Chlorella vulgaris
Details on test organisms:
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:
24 °C ±2°C.
pH:
6.7 to 7.4
Nominal and measured concentrations:
6.25 mg/L,12.5 mg/L,25 mg/L,50 mg/L,100 mg/L,200 mg/L.All the six concentration were in geometric series spaced by a factor of 2.
Details on test conditions:
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
Key result
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: calculated from equation through probit analysis
Details on results:
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.

Table 1: Showing the average cell count using Haemocytometer of the experimental flasks at an equal interval of 24hrs, 48hrs and 72hrs

Validity criteria fulfilled:
yes
Conclusions:
After 72 hours of exposure to Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex(CAS No12225-21-7) to various nominal test concentration, EC50 calculated from equation through probit analysis was found to be >100 mg/L.
Executive summary:

The effect of test item Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1Hpyrazole-3-carboxylic acid complex, (CAS No. 12225-21-7) 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 6.25 mg/L,12.5 mg/L,25 mg/L,50 mg/L,100 mg/L,200 mg/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 >200 mg/L. It can be concluded from the value that the Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1Hpyrazole-3-carboxylic acid complex, (CAS No. 12225-21-7) is not toxic to the aquatic algae and can be considered as "not classified" as per the classifiecation criteria for aquatic environment.

Description of key information

The effect of test item Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1Hpyrazole-3-carboxylic acid complex, (CAS No. 12225-21-7) 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 6.25 mg/L,12.5 mg/L,25 mg/L,50 mg/L,100 mg/L,200 mg/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 >200 mg/L. It can be concluded from the value that the Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1Hpyrazole-3-carboxylic acid complex, (CAS No. 12225-21-7) is not toxic to the aquatic algae and can be considered as "not classified" as per the classifiecation criteria for aquatic environment.

Key value for chemical safety assessment

EC50 for freshwater algae:
200 mg/L

Additional information

Data is from UERL 2017, exeprimental report followed standard gudeline suggests that The effect of test item Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1Hpyrazole-3-carboxylic acid complex, (CAS No. 12225-21-7) 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 6.25 mg/L,12.5 mg/L,25 mg/L,50 mg/L,100 mg/L,200 mg/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 >200 mg/L. It can be concluded from the value that the Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1Hpyrazole-3-carboxylic acid complex, (CAS No. 12225-21-7) is not toxic to the aquatic algae and can be considered as "not classified" as per the classifiecation criteria for aquatic environment.