Registration Dossier

Ecotoxicological information

Endpoint summary

Administrative data

Description of key information

Short term toxicity to fish:

An experiment was conducted to determine the short term toxicity of test chemical to fish. In this study test organism used was Danio rerio (previous name: Brachydanio rerio). The nominal concentrations of test chemical used were of 0, 5.66 or 8.0 mg/l. The test chemical purity was reported to be 30% active ingredient The experiment was conducted in static condition and test chemical was exposed to fish for 96 h. All fish exposed to 8.0 mg/L died within the first 24 hours of exposure. The medial lethal concentration (EC50) for test chemical on Danio rerio after 96 h was observed to be 2.0 mg/l. Since the test chemical is readily biodegradable in nature test chemical is considered to be non toxic and cannot be Classified as per CLP regulation.

Long term toxicity to fish:

An experiment was conducted to determine the long term toxicity of test chemical by using OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study). In this study test organism used was Oncorhynchus mykiss. The no effect concentration (NOEC) and low effect concentration (LOEC) after 28 days was observed to be 0.16 mg/l and 0.5 mg/l respectively. By considering the no effect concentration (NOEC) value test chemical is considered to be toxic and can be classified as aquatic chronic 3 for long term toxicity to fish.

Short term toxicity to aquatic invertebrate:

An experiment was conducted to determine the short term toxicity of test chemical to aquatic invertebrates according to theProtocol number 202, part I, 24 h EC50 Acute Immobilization Test, of the 4th of April 1984, from theOECD Guideline for Testing of Chemicals. The test organisms used in the study was Daphnia magna. The experiment was conducted in static condition and waster used was 50% dechlorinated water and 50% distilled water. The nominal concentrations of test chemical used were 0 (control), 0.5, 1, 2, 4, 8 and 16 mg/l. The test chemicalwas administered in a single dose per concentration level, dissolved in the water held in the test tubes, which was similar to that used in the breeding period. The daphnia were then transferred to the vessels.

The dilution water was aerated to saturation level prior to introduction of the test substance to ensure that the oxygen level did not fall below 60% of the saturation value.Values for pH ranged from 8.33 –8.49, temperature ranged between 21.9 – 22.4°C and dissolved oxygen ranged between 67 – 100%.The Daphnia were observed at the initiation of study and after 24 and 48 hours exposure. Observations included a determination of the number of immobilized animals.The test chemical caused 0% immobilization at the 0.5 mg/l concentration. At concentrations of 1 mg/l and 2 mg/l, 2 and 3 daphnia, respectively, out of a total of 40, were found immobile. The 4 mg/l and 8 mg/l concentrations caused immobilization in 5 and 9 daphnia, respectively, out of a total of 20. All the animals treated at the 16 mg/l concentration were observed to be immobile. 1.7% immobilization was recorded in the Control group.

The statistical method used in this study was Litchfield and Wilcoxon Method.

The effect concentration at which 50% immobilization (EC50) of daphnia was observed was determined to be 6.40 mg/l (confidence limit 4.57-8.96). Since the test chemical is readily biodegradable in nature test chemical is considered to be non toxic and cannot be classified as per CLP regulation.

Long term toxicity to aquatic invertebrate:

An experiment was conducted to determine the Long term toxicity of test chemical to aquatic invertebrates according toOECD 202 Daphnia Reproduction Test.  The test organism used in this study was Daphnia magna. The experiment was conducted in semi-static condition using fresh water. The test chemical was exposed to Daphnia Magna for 21 days. The no observed effected concentration (NOEC) and low observed effect concentration (LOEC) was observed to be 0.9 mg/l and 3.6 mg/l respectively. By considering the LOEC value test chemical is considered to be non toxic to aquatic invertebrates and cannot be classified as per CLP regulation

Toxicity to aquatic algae and cyanobacteria:

Toxicity of aquatic algae and cyanobacteria was evaluated for test chemical Cocamidopropyl Betaine on marine microalgae Ulva lactuca.

 

Prior to experimentation, thalli were maintained for 3 days in Perspex aquaria containing 5 l of aerated, filtered , seawater (salinity = 33.8) in a temperature-controlled culture room set at 15° (representative of local, summer seawater temperatures) under a irradiance of on a 12:12 h light: dark photoperiod.Approximately 24 h prior to commencing the experiments, discs of 16 mm diameter were cut from the central portion of ca. 30 individual thalli using a plastic cork borer and left to recover from any resulting trauma. From the pooled discs, two or three were randomly selected and transferred to individual glass beakers containing 100 ml of treatment solutions.

 

The nominal concentration selected for the experiment were 0, 5, 10, 15, 20, 25, 30, 35 and 40 mg/l. Data were analysed by either Repeat Measure Analysis of Variance (ANOVA) using SPSS (v.15.0) or by one-way or two-way ANOVA using Minitab (v. 15).The 50% effect on biomass of marine microalgae was observed in the concentration 30 mg/l after 48 h.

Though , according to the above concentration the test material can be considered as toxic to aquatic environment and can be classified as acute aquatic – 3, but the substance is readily biodegradable in aquatic environment. Hence, the test material can not be classified as per CLP classification.

Toxicity to microorganism:

Toxicity of test material Cocamidopropyl Betaine was evaluated for its efficiency in cell multiplication and oxygen consumption inhibition of test organism Pseudomonas putida for 16 hrs. No adverse effect of test material was observed on test organism Pseudomonas putida for its cell multiplication and oxygen consumption inhibition. the reported EC0 was >3000 mg/l

Additional information

Short term toxicity to fish:

Various studies have been conducted to determine the toxicity of test chemical to fish from different sources and their results are summarized below. 

 

In first study an experiment was conducted to determine the short term toxicity of test chemical to fish. In this study test organism used was Danio rerio (previous name: Brachydanio rerio). The nominal concentrations of test chemical used were of 0, 5.66 or 8.0 mg/l. The test chemical purity was reported to be 30% active ingredient The experiment was conducted in static condition and test chemical was exposed to fish for 96 h. All fish exposed to 8.0 mg/L died within the first 24 hours of exposure. The medial lethal concentration (EC50) for test chemical on Danio rerio after 96 h was observed to be 2.0 mg/l. 

 

In another study the experiment was conducted to determine the short term toxicity of test chemical to fish by using OECD Guideline 203 (Fish, Acute Toxicity Test). In this study test organism used was Danio rerio (previous name: Brachydanio rerio) whose Length at study initiation of study was 2.4 and 3.0 cm long and Acclimation period was of 12 days. In study two groups were made, each containing 10 animals. Ten Control fish were also included. Animals were placed in 84-liter glass tanks containing 50 liters of de chlorinated drinking water.

After exposure of test chemical of concentration 8.0 mg/l to fish 100% mortality was observed during the first 24 hours. Before death, they showed a decrease in mobility and breathing difficulties. Two of the animals remained on the surface of the aquarium and the others at the bottom. The 5.66 mg/l concentration did not cause the death of any of the treated animals. Only a slight decrease in mobility was noted between 3 and 24 hours after treatment. No mortality was observed in the control group. The medial lethal effect concentration was observed to be 6.73 mg/l after 96 h of exposure. 

 

The last experimental study was conducted to determine the short term toxicity of test chemical. In this study test organism used was Cyprinus carpio. The active ingredient of test chemical was reported29.6% .The nominal concentrations of test chemical used were 0, 1.0, 1.7, 3.0, 5.0, and 9 mg/L. The experiment was conducted under semi-static condition for 96 h. The median lethal concentration (LC 50 value) of test chemical on Cyprinus carpio was observed to be 1.9 mg/l after 96 h.

 

By considering results of all the studies mentioned above the LC50 value was determined to be in range 2.0 mg/L to 6.73 mg/l. Since the test chemical is readily biodegradable in nature test chemical is considered to be non toxic and cannot be classified as per CLP regulation.

Long term toxicity to fish:

An experiment was conducted to determine the long term toxicity of test chemical by using OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study). In this study test organism used was Oncorhynchus mykiss. The no effect concentration (NOEC) and low effect concentration (LOEC) after 28 days was observed to be 0.16 mg/l and 0.5 mg/l respectively. By considering the no effect concentration (NOEC) value test chemical is considered to be toxic and can be classified as aquatic chronic 3 for long term toxicity to fish.

Short term toxicity to aquatic invertebrate:

Different studies have been conducted to determine the toxicity of test chemical to aquatic invertebrates from various sources and their results are summarized below. 

 

In first study an experiment was conducted to determine the short term toxicity of test chemical to aquatic invertebrates according to the Protocol number 202, part I, 24 h EC50 Acute Immobilization Test, of the 4th of April 1984, from the OECD Guideline for Testing of Chemicals. The test organisms used in the study was Daphnia magna. The experiment was conducted in static condition and waster used was 50% dechlorinated water and 50% distilled water. The nominal concentrations of test chemical used were 0 (control), 0.5, 1, 2, 4, 8 and 16 mg/l. The test chemical was administered in a single dose per concentration level, dissolved in the water held in the test tubes, which was similar to that used in the breeding period. The daphnia were then transferred to the vessels.

The dilution water was aerated to saturation level prior to introduction of the test substance to ensure that the oxygen level did not fall below 60% of the saturation value. Values for pH ranged from 8.33 –8.49, temperature ranged between 21.9 – 22.4°C and dissolved oxygen ranged between 67 – 100%.The Daphnia were observed at the initiation of study and after 24 and 48 hours exposure. Observations included a determination of the number of immobilized animals. The test chemical caused 0% immobilization at the 0.5 mg/l concentration. At concentrations of 1 mg/l and 2 mg/l, 2 and 3 daphnia, respectively, out of a total of 40, were found immobile. The 4 mg/l and 8 mg/l concentrations caused immobilization in 5 and 9 daphnia, respectively, out of a total of 20. All the animals treated at the 16 mg/l concentration were observed to be immobile. 1.7% immobilization was recorded in the Control group.

The statistical method used in this study was Litchfield and Wilcoxon Method.

The effect concentration at which 50% immobilization (EC50) of daphnia was observed was determined to be 6.40 mg/l (confidence limit 4.57-8.96).

 

In another study an experiment was conducted to determine the short term toxicity of test chemical to aquatic invertebrates according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) and EU Method C.2 (Acute Toxicity for Daphnia). The test organism used in the study was Daphnia Magna which were laboratory bred and less than 24 hours old at test initiation. The nominal concentrations of test chemical were0 (control), 6.25, 12.5, 25, 50 and 100 mg/l. Dilution water was reconstituted water prepared according to the EEC Directive. The pH of the water was adjusted to 8.1 and the dissolved oxygen was 8.1 mg/l. The experiment was conducted in static condition in50-ml beakers containing 20 ml of test solution with 10 Daphnia Magna per beaker. Experiments were run in duplicate. The concentration of the test chemicals at the beginning and at the end of the test was analyzed for the control and for the 6.25, 25, and 100 mg/l test solutions. In addition, stability analyses gave values 95.5 and 85.2% of nominal for a 100 mg/l solution at the beginning and end of the test. The statistical method used in this study was Logit model described by Cox, D. R. 1977. Analysis of binary data. Methuen & Co., Ltd. Daphnids in the control group showed no adverse effects during the exposure.

After 48 h exposure of test chemical to Daphnia magna was the Effect concentration (EC50) at which 50 % immobilization observed was determined to be 21.5 mg/l (95% confidence limits: 16.1 – 28.1 mg/l). EC0 and EC100 value were determined to be 5.3 mg/l and 89.3 mg/l respectively after exposure of test chemical to Daphnia magna for 48 h.

 

By considering results of all the studies mentioned above the EC50 value was determined to be in range 6.40 mg/L to 21.5 mg/l. Since the test chemical is readily biodegradable in nature test chemical is considered to be non toxic and cannot be classified as per CLP regulation.

Long term toxicity to aquatic invertebrate:

An experiment was conducted to determine the Long term toxicity of test chemical to aquatic invertebrates according toOECD 202 Daphnia Reproduction Test.  The test organism used in this study was Daphnia magna. The experiment was conducted in semi-static condition using fresh water. The test chemical was exposed to Daphnia Magna for 21 days. The no observed effected concentration (NOEC) and low observed effect concentration (LOEC) was observed to be 0.9 mg/l and 3.6 mg/l respectively. By considering the LOEC value test chemical is considered to be non toxic to aquatic invertebrates and cannot be classified as per CLP regulation

Toxicity to aquatic algae and cyanobacteria:

Different studies have been conducted to determine the toxicity of test chemical to aquatic algae from different sources and their results are summarized below.                                                                                   

 

The first study was reviewed from the ECotoxiccology journal (2011) in this an experiment was conducted to determine the toxicity of test chemical to aquatic algae. In this the test organisms used was marine microalgae Ulva lactuca. Before initiation of experiment the thalli were maintained for 3 days in Perspex aquaria containing 5 l of aerated, filtered , seawater (salinity = 33.8) in a temperature-controlled culture room set at 15° (representative of local, summer seawater temperatures) under a irradiance of on a 12:12 h light: dark photoperiod. Approximately 24 h prior to commencing the experiments, discs of 16 mm diameter were cut from the central portion of ca. 30 individual thalli using a plastic cork borer and left to recover from any resulting trauma. From the pooled discs, two or three were randomly selected and transferred to individual glass beakers containing 100 ml of treatment solutions.

The nominal concentration selected for the experiment were 0, 5, 10, 15, 20, 25, 30, 35 and 40 mg/l. Data were analysed by either Repeat Measure Analysis of Variance (ANOVA) using SPSS (v.15.0) or by one-way or two-way ANOVA using Minitab (v. 15). The Effect concentration at which 50% effect on biomass of marine microalgae was observed was 30 mg/l after 48 h.

 

In another study an experiment was conducted to determine the toxicity of test chemical to aquatic algae according to the OECD Guideline 201 (Alga, Growth Inhibition Test). In this experiment test organism used was Scenedesmus subspicatus. The nominal concentration sof test chemical used were 0 (control), 0.32, 1.0, 3.2, 10, 32, and 100 mg/l. The experiment was conduceted in ststic condition for 72 h at temperature 23 ± 2 °C. Four replicate test flasks were run at each treatment level. Tests were run under continuous lighting of 35 – 70 μE/m2*s. Endpoints were determined for algal growth rate and algal cell density. The EC0, EC10, and EC50 were calculated for rate and cell density. The effect concentrations of biomass and growth rate on green algae after 72 h was observed to be 30 mg/l and 48 mg/l respectively.

 

Next study was reviewed from secondary source in this an experiment was conducted to determine the toxicity of test chemical to aquatic algae according to the DIN 38412, Part 9. In this experiment test organism used was Scenedesmus subspicatus -Strain: SAG 8681.The nominal concentration of test chemical used in this were 0 (control), 0.01, 0.03, 0.1, 0.3, 1.0, 3.0 and 10 mg/L. Each experimental group was replicated three times. Test vessels were 300-ml Erlenmeyer flasks holding 100 ml of test solution. At the beginning of the test, flasks were inoculated with 1.0 ml of algal cell inoculum to achieve a concentration in each flask of 1 × 104 cells/ml. Flasks were placed under continuous lighting of 2000 lux and continuously shaken at 120 rpm by means of an orbital shaker. At 24, 48, 72 and 96 hours, a sample from each flask was taken and the density of algal cells in the sample (cells/ml) was measured using an electronic particle counter (Coulter-Counter). Cell densities were converted to growth rates and growth rates were used in the calculation of EC values.

The EC50 Value of test chemical was determined to be 1.84 mg/L after exposure of test chemical for 96 h.

 

By considering results of all the studies mentioned above the EC50 value was determined to be in range 1.84 mg/L to 48 mg/l. Since the test chemical is readily biodegradable in nature test chemical is considered to be non toxic and cannot be classified as per CLP regulation.

Toxicity to microorganism:

Different studies have been conducted to determine the toxicity of test chemical to microorganism and their results are summarized below.                                                                                   

 

The first study was conducted to determine the toxicity of test chemical on microorganism, in this the microorganism used was Pseudomonas putida. The efficiency in cell multiplication and oxygen consumption inhibition of test organism was observed. The effect concentration at which no effect was observed was determined to be >3000 mg/l after exposure of test chemical for 16 h.

 

In another study the experiment was conducted to determine the toxicity of test chemical to microorganisms. The test organisms used were Pseudomonas aeruginosa, Staphylococcus epidermis, Bacillus albicans and Aspergillus niger. The test chemical was exposed to these microorganisms for 24 h. The concentrations at which no effects (EC0) were observed was determined to be>100 mg/L onPseudomonas aeruginosa. The concentration of test chemical at which 10% inhibitions (EC10) occurs was determined to be 2.5mg/L on Staphylococcus epidermis and 20 mg/L on Bacillus albicans and Aspergillus niger.

 

By considering results of both the studies mentioned above the EC0 value was determined to be in range >100 mg/L to 3000 mg/L.