1-dodecylpyridinium chloride

Endpoint summary

Description of key information

Short term toxicity to fish

Short term toxicity to fish was conducted for 96 hrs for assessing the effect of test chemical. The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.71 g and average length of 1.75 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.6 mg/l, pH 7.5, water temperature 24.2°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. The test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous one hour of stirring for achieving test concentrations of 0.78 mg/L,1.56 mg/L,3.125 mg/L,6.25 mg/L,12.5 mg/L, respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 24.2°C, pH 7.3, hardness of water 154.2 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control. Test fishes were moving slowly as compared to control. On the basis of effect of test chemical on mortality of the test organism, the median lethal concentration (LC50 (96 h)) was determined to be >6.25 to <12.5 mg/l. Thus, based on the LC50 value, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates

This study was designed (as per OECD 202, adopted in 2004) to assess the acute toxicity of test chemical following exposure of daphnids up to 48h by static method. The test daphnids were acclimatized 48 hours prior to the test chemical exposure. After exposure on day 0, daphnids were observed for immobilization at 24 and 48 h. M7 medium was used as control, and the same was used for test chemical formulation . 100 mL glass beakers having a solution volume of 30 mL were used in the test. . Main study (using a spacing factor of 2) was conducted using 0 (control), 0.125, 0.25, 0.5, 1 and 2 mg/L concentrations. 2 replicates/concentration having 5 daphnids/replicate was used for the main study. UV-Visible spectrophotometer method was used for active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 h and after 48 hours of exposure, which were found in acceptable range. Hence the results were based on nominal concentration since the deviation in the initial measured concentration didn’t exceed 20%. Environmental parameters such as pH (7.3 -7.6), temperature (20-21°C), dissolve oxygen (7.4 - 8.1 mg/L), hardness (225 mg CaCO3/L), light intensity of 1086 lux, photoperiod (16 h light- 8 h dark) was maintained in acceptable range throughout the test. Thus, fulfilling the validity criteria. Feed was not provided during the test. Normal behavioural response and no immobilization (0% mortality) were observed up to 48 h followed by control groups but 40%, 60%, 100%, 100% and 100% immobilisation were observed in the test concentrations of 0.125, 0.25, 0.5, 1 and 2 mg/L, respectively. The 48-h EC50 of test chemical to daphnid, Daphnia magna are 0.16 mg/l. The 24-h EC50 of reference item (Potassium dichromate) to daphnid, Daphnia magna(found to be in acceptable range) is 1.33 mg/L. Hence, the results of the test with reference item establish the acceptability of the test system response, test procedures followed, and results obtained with test item. Thus, test chemical was considered as toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be classified in 'aquatic acute/chronic category 1' as per the CLP classification criteria.

Long term toxicity to aquatic invertebrates

Long term toxicity test was performed on M.Mercenaria (Clam) larvae.10,000 to 15,000 larvae reared to the straight hinge larval stage. The cultures were fed a mixture of flagellated algal food and continued exposed to the test chemical for 12days. Effects on cultures based on survival and growth was determined.

Based on the results it was concluded that the endpoint for long term toxicity to aquatic invertebrates to test material was found to be LOEC at 0.25 mg/l and 0.050 mg/l concentrations based on survival and growth of M.Mercenaria (Clam) larvae.This LOEC value suggest that the test chemical was toxic to aquatic invertebrates in chronic exposure.

Toxicity to aquatic algae and cyanobacteria

A freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae Pseudokirchneriella subcapitata. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. OECD medium composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test solution was prepared by dissolving 250 mg of test chemical in 250 ml of OECD medium to get the final concentration of 100 mg/L. To have a better growth and visibility of cells, the initial cell count of the culture was kept 10,000 cells/mL. Further, exposure concentrations of 0, 3, 4.8, 7.68, 12.29 and 19.66 mg/L mg/l (factor of 1.6), respectively was from the stock test concentrations. Test vessel were placed in orbital shaking incubator for 72 hrs at a temperature of 21 to 24 ± 2°C and with a continuous uniform illumination of 6000 -10000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. The cultures were counted and observed daily with the help of a microscope. Potassium dichromate (K2Cr2O7) was used as a reference substance. The 72 hr EC50 value of the reference substance was determined to be 1.57 mg/l. The biomass in the control vessel have increased exponentially by a factor of 16, the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% (i.e., reported to be 0.838%) and the mean coefficient of variation of specific growth rate was not exceeded 35% (i.e., reported to be 4.33%), respectively thus fulfilling the validity criterion. As the concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. On the basis of growth rate of the test organism, the 72 hr median effect concentration (ErC50) value was determined to be 5.94 mg/l (nominal conc.). Thus, test chemical was considered as toxic to aquatic algae at environmental relevant concentrations and hence, considered to be classified in 'aquatic chronic category 2' as per the CLP classification criteria.

Toxicity to microorganisms

The antibacterial effect of cationic surfactant against the pure culture of phosphate (P)-accumulating bacteriumAcinetobacter juniiwas investigated. The bacteria were pregrown on nutrient agar for 20 h at 30±0.1°C to obtain log phase cell culture. The biomass was then resuspended in 10 mL of 0.3% NaCl and 1 mL of mixed suspension was distributed in series of bottles containing different concentration of test material per 100 mL of simulative wastewater. The concentrations used were 10e-4, 10e-5, 10e-6, 10e-7 mol/L. The bottles were sealed with a sterile gum cap and thereafter purged with filtered air during a 24-hour incubation in a temperature-controlled water bath shaker at 30±0.1°C and 70 rpm. The broth bottle syringe method of YTT (Yeast toxicity test) was used to compare the toxicity obtained againstA. juniiwith the standardized toxicity test. Yeast used wasS. cerevisiae.The control contained no test chemical. The bottles were sealed with a sterile gum cap and thereafter purged with filtered air during a 24-hour incubation in a temperature-controlled water bath shaker at 30±0.1°C and 70 rpm.The EC50 values of the test material (D for growth inhibition of phosphate (P)-accumulating bacteriumAcinetobacter juniiwas 1.4±0.5 x 10-6 mol/ L (0.3974 mg/L). Thus based on the above data, it can be considered that the substance test material is toxic to microorganisms.

Additional information

Short term toxicity to fish

Experimental key and supporting study of the test chemical were reviewed for short term toxicity to fish end point which are summarized as below:

 

In an experimental study from study report, short term toxicity to fish was conducted for 96 hrs for assessing the effect of test chemical. The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.71 g and average length of 1.75 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.6 mg/l, pH 7.5, water temperature 24.2°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. The test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous one hour of stirring for achieving test concentrations of 0.78 mg/L,1.56 mg/L,3.125 mg/L,6.25 mg/L,12.5 mg/L, respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 24.2°C, pH 7.3, hardness of water 154.2 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control. Test fishes were moving slowly as compared to control. On the basis of effect of test chemical on mortality of the test organism, the median lethal concentration (LC50 (96 h)) was determined to be >6.25 to <12.5 mg/l. Thus, based on the LC50 value, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

 

Another short term Toxicity test according to Polish Standards (PN-90/C-04610/03) was conducted for test material. Lebistes reticulatus (Guppy) of length 8-12 mm was used as a test organism. Test fishes were taken before the occurrence of sex dimorphism. Minimum 5 different concentrations (conc. spacing of 1.8) were taken for the study. Three parallel tests were performed in 500 ml container with seven organisms for a period of 96 hrs. For estimation of LC50, log-probit method was used. The 96 hr lethal concentrations (LC50) value was determined to be 11.5 mg/l. Thus, based on the LC50 value, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

 

On the basis of the above results, it can be concluded that the test chemical was considered to be toxic to aquatic fishes and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates

This study was designed (as per OECD 202, adopted in 2004) to assess the acute toxicity of test chemical following exposure of daphnids up to 48h by static method. The test daphnids were acclimatized 48 hours prior to the test chemical exposure. After exposure on day 0, daphnids were observed for immobilization at 24 and 48 h. M7 medium was used as control, and the same was used for test chemical formulation . 100 mL glass beakers having a solution volume of 30 mL were used in the test. . Main study (using a spacing factor of 2) was conducted using 0 (control), 0.125, 0.25, 0.5, 1 and 2 mg/L concentrations. 2 replicates/concentration having 5 daphnids/replicate was used for the main study. UV-Visible spectrophotometer method was used for active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 h and after 48 hours of exposure, which were found in acceptable range. Hence the results were based on nominal concentration since the deviation in the initial measured concentration didn’t exceed 20%. Environmental parameters such as pH (7.3 -7.6), temperature (20-21°C), dissolve oxygen (7.4 - 8.1 mg/L), hardness (225 mg CaCO3/L), light intensity of 1086 lux, photoperiod (16 h light- 8 h dark) was maintained in acceptable range throughout the test. Thus, fulfilling the validity criteria. Feed was not provided during the test. Normal behavioural response and no immobilization (0% mortality) were observed up to 48 h followed by control groups but 40%, 60%, 100%, 100% and 100% immobilisation were observed in the test concentrations of 0.125, 0.25, 0.5, 1 and 2 mg/L, respectively. The 48-h EC50 of test chemical to daphnid, Daphnia magna are 0.16 mg/l. The 24-h EC50 of reference item (Potassium dichromate) to daphnid, Daphnia magna(found to be in acceptable range) is 1.33 mg/L. Hence, the results of the test with reference item establish the acceptability of the test system response, test procedures followed, and results obtained with test item. Thus, test chemical was considered as toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be classified in 'aquatic acute/chronic category 1' as per the CLP classification criteria.

Long term toxicity to aquatic invertebrates

In a peer reviewed journal study for 21 days was described for two aquatic invertebrates, M.Mercenaria (Clam) and C. Virginica(Oysters) larvae

Long term toxicity test was performed on M.Mercenaria (Clam) larvae.10,000 to 15,000 larvae reared to the straight hinge larval stage. The cultures were fed a mixture of flagellated algal food and continued exposed to the test chemical for 12days. Effects on cultures based on survival and growth was determined.

Based on the results it was concluded that the endpoint for long term toxicity to aquatic invertebrates to test material was found to be LOEC at 0.25 mg/l and 0.050 mg/l concentrations based on survival and growth of M.Mercenaria (Clam) larvae.This LOEC value suggest that the test chemical was toxic to aquatic invertebrates in chronic exposure.

Long term toxicity test was performed on C. Virginica(Oysters) larvae.10,000 to 15,000 larvae reared to the straight hinge larval stage. The cultures were fed a mixture of flagellated algal food and continued exposed to the test chemical for 14 days. Effects on cultures based on survival and growth was determined.

Based on the results it was concluded that the endpoint for long term toxicity to aquatic invertebrates to test material was found to be LOEC at 0.25 mg/l and 0.050 mg/l concentrations based on survival and growth of C. Virginica (Oysters) larvae.This LOEC value suggest that the test chemical was toxic to aquatic invertebrates in chronic exposure. Based on the above effect concentration we can consider the test material to be classiffied as aquatic chronic 3

Toxicity to aquatic algae and cyanobacteria

A freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae Pseudokirchneriella subcapitata. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. OECD medium composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test solution was prepared by dissolving 250 mg of test chemical in 250 ml of OECD medium to get the final concentration of 100 mg/L. To have a better growth and visibility of cells, the initial cell count of the culture was kept 10,000 cells/mL. Further, exposure concentrations of 0, 3, 4.8, 7.68, 12.29 and 19.66 mg/L mg/l (factor of 1.6), respectively was from the stock test concentrations. Test vessel were placed in orbital shaking incubator for 72 hrs at a temperature of 21 to 24 ± 2°C and with a continuous uniform illumination of 6000 -10000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. The cultures were counted and observed daily with the help of a microscope. Potassium dichromate (K2Cr2O7) was used as a reference substance. The 72 hr EC50 value of the reference substance was determined to be 1.57 mg/l. The biomass in the control vessel have increased exponentially by a factor of 16, the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% (i.e., reported to be 0.838%) and the mean coefficient of variation of specific growth rate was not exceeded 35% (i.e., reported to be 4.33%), respectively thus fulfilling the validity criterion. As the concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. On the basis of growth rate of the test organism, the 72 hr median effect concentration (ErC50) value was determined to be 5.94 mg/l (nominal conc.). Thus, test chemical was considered as toxic to aquatic algae at environmental relevant concentrations and hence, considered to be classified in 'aquatic chronic category 2' as per the CLP classification criteria.

Toxicity to microorganisms

Experimental study of the test chemical and supporting study for its read across chemical were reviewed for toxicity to microorganisms end point which are summarized as below:

The antibacterial effect of cationic surfactant against the pure culture of phosphate (P)-accumulating bacterium Acinetobacter junii was investigated. The bacteria were pregrown on nutrient agar for 20 h at 30±0.1°C to obtain log phase cell culture. The biomass was then resuspended in 10 mL of 0.3% NaCl and 1 mL of mixed suspension was distributed in series of bottles containing different concentration of test material per 100 mL of simulative wastewater. The concentrations used were 10e-4, 10e-5, 10e-6, 10e-7 mol/L. The bottles were sealed with a sterile gum cap and thereafter purged with filtered air during a 24-hour incubation in a temperature-controlled water bath shaker at 30±0.1°C and 70 rpm. The broth bottle syringe method of YTT (Yeast toxicity test) was used to compare the toxicity obtained against A. junii with the standardized toxicity test. Yeast used was S. cerevisiae. The control contained no test chemical. The bottles were sealed with a sterile gum cap and thereafter purged with filtered air during a 24-hour incubation in a temperature-controlled water bath shaker at 30±0.1°C and 70 rpm. The EC50 values of the test material (D for growth inhibition of phosphate (P)-accumulating bacterium Acinetobacter junii was 1.4±0.5 x 10-6 mol/ L (0.3974 mg/L). Thus based on the above data, it can be considered that the substance test material is toxic to microorganisms.

 

The above data was further supported by data from peer reviewed journal for read across substance, the antibacterial effect of cationic surfactant against the pure culture of phosphate (P)-accumulating bacterium Acinetobacter juniiwas investigated. The bacteria were pregrown on nutrient agar for 20 h at 30±0.1°C to obtain log phase cell culture. The biomass was then resuspended in 10 mL of 0.3% NaCl and 1 mL of mixed suspension was distributed in series of bottles containing different concentration of Test material per 100 mL of simulative wastewater. The concentrations used were 10-4, 10-5, 10-6, 10-7mol L-1. The bottles were sealed with a sterile gum cap and thereafter purged with filtered air during a 24-hour incubation in a temperature-controlled water bath shaker at 30±0.1°C and 70 rpm. The broth bottle syringe method of YTT (Yeast toxicity test) was used to compare the toxicity obtained against A. Junii with the standardized toxicity test. Yeast used was S. cerevisiae. The control contained no test chemical. The bottles were sealed with a sterile gum cap and thereafter purged with filtered air during a 24-hour incubation in a temperature-controlled water bath shaker at 30±0.1°C and 70 rpm. The estimated EC50 values of the test material for growth inhibition of phosphate (P)-accumulating bacterium Acinetobacter junii was 4.9±1.3 x 10-7 mol L-1. Thus based on the above data, it can be considered that the test substance is toxic to microorganisms.

On the basis of the available information of aquatic toxicity studies, it can be concluded that the test chemical was considered to be toxic to aquatic organismsat environmental relevant concentrationsand hence, considered to be classified in ‘aquatic acute/chronic category 1’ as per the CLP classification criteria.