Methyltrioctylammonium chloride

Endpoint summary

Description of key information

Short term toxicity to fish:

Study was conducted for the evaluation of acute effect of test chemicalMethyltrioctylammonium chloride on the mortality rate of zebrafish under static system for 144 hrs. Toxicant solutions were prepared by pipetting into wide mouthed Erlen-Meyer flasks appropriate amounts (0-5.6 ml) of toxicant stock solutions made up in acetone. After evaporation of the acetone just to dryness at room temperature, 1000 ml of premade aerated SRW2 (Standard reference water) was added to each flask. A solvent control (maximal amount of acetone evaporated just to dryness + 1000 ml diluent) and an ordinary control (1000 ml diluent) were included in the entire test procedure. All solutions were allowed to stand over night at the test temperature (25°C).

5 Juvenile zebrafish were used. Effect measured in the interval of 3, 6, 24, 48, 72, 96, 120 and 144 hr. Effect on the mortality were measured and LC50 noted by probit analysis. As the zebrafish exposed with the test chemical Methyltrioctylammonium chloride for 144 hrs, mortality were observed. The effect were measured at 24, 48, 96 and 144 hrs. The LC50 after 96 and 144 hrs observed at 0.137 mg/l. whereas after 24 and 48 hrs LC50 ranges from 0.108 MG/L to 0.131 MG/L. Thus based on the LC50 value, chemical consider to be toxic and can be consider to be classified as aquatic acute 1 / chronic 2 category as per the CLP classification criteria.

Short term toxicity to invertebrates:

Aim of this study was to assess the short term toxicity of Methyltrioctylammonium chloride to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs. The stock solution 20 mg/l was prepared by dissolving colourless dense liquid in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample in reconstituted water. 0, 0, 0.006, 0.012, 0.025, 0.050, 0.100, 0.200 mg/l concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance Methyltrioctylammonium chloride, in Daphnia magna was determined to be 0.031 mg/L on the basis of mobility inhibition effects in a 48 hour study. Thus EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic acute 1 / chronic 1 category as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of chemical test chemical Methyltrioctylammonium chloride when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD guideline 201. The stock solution 100 mg/l was prepared by dissolving colourless dense liquid in OECD growth medium. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Various concentrations 0, 0.05, 0.10, 0.20, 0.40, 0.80 mg/l were used. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. Effect on the growth of algae was determine after an exposure period of 72 hrs. The median effective concentration (EC50) for the test substance Methyltrioctylammonium chloride, in algae was determined to be 0.56 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Thus based on the EC50 value, it indicates that the substance is likely to be hazardous to aquatic algae and can be consider to be classified as aquatic acute1 / chronic 2 category as per the CLP classification criteria.

Toxicity to microorganism:

Bioassay for the inhibition of growth of Cellulomonas sp. was performed for the test material Methyltrioctylammonium chloride.Test organism were inoculated in a synthetic medium prepaired by adding glucose 6 mM; (NH4)2HP04 4 mM; K2HPO4 6 mM; KH2P04 10 mM; KCI 7 mM; EDTA 1 mM; vitamins and trace elements. Inoculums were prepared from 100 cm³cultures grown in 250 cm³Erlenmeyer flasks on a rotary shaker.

Initial cell concentration was recorded to be 10⁷cells cm^-3.The experiments were performed in 100cm³Erlenmeyer flasks equipped with a side tube, making direct measurements possible. The flasks contained 50cm³medium. Incubation was made on a rotary shaker, 120 rev min^-1. Growth was measured turbid metrically at 610 nm.The exponential growth rate was determined from the linear part of the growth curves by means of linear regression analysis.

 

The Effect concentration (EC50) of test material Methyltrioctylammonium chloride on Cellulomonas sp. was observed to be in the range 0.2 – 0.4 mg/l for the exponential growth rate and the effect concentration (EC50) on biomass was observed to be in the range 0.1 – 0.2 mg/l.Based on the above effect concentration test material can be considered toxic to microorganisms.

Additional information

Short term toxicity to fish:

Summarized result from peer reviewed journals for the toxicity of test chemical on the mortality and embryo development of fishes was studied and mention as follows:

In the first study from peer reviewed journal, Study was conducted for the evaluation of acute effect of test chemicalMethyltrioctylammonium chloride on the mortality rate of zebrafish under static system for 144 hrs. Toxicant solutions were prepared by pipetting into wide mouthed Erlen-Meyer flasks appropriate amounts (0-5.6 ml) of toxicant stock solutions made up in acetone. After evaporation of the acetone just to dryness at room temperature, 1000 ml of premade aerated SRW2 (Standard reference water) was added to each flask. A solvent control (maximal amount of acetone evaporated just to dryness + 1000 ml diluent) and an ordinary control (1000 ml diluent) were included in the entire test procedure. All solutions were allowed to stand over night at the test temperature (25°C). 5 Juvenile zebrafish were used. Effect measured in the interval of 3, 6, 24, 48, 72, 96, 120 and 144 hr. Effect on the mortality were measured and LC50 noted by probit analysis. As the zebrafish exposed with the test chemical Methyltrioctylammonium chloride for 144 hrs, mortality were observed. The effect were measured at 24, 48, 96 and 144 hrs. The LC50 after 96 and 144 hrs observed at 0.137 mg/l. whereas after 24 and 48 hrs LC50 ranges from 0.108 MG/L to 0.131 MG/L. Thus based on the LC50 value, chemical consider to be toxic and can be consider to be classified as aquatic acute 1 / chronic 2 category as per the CLP classification criteria.

First study was supported by the second from peer reviewed journal 1981. Embryo-larval toxicity by the test chemical Methyltrioctylammonium chloride to zebrafish was studied under the flow-through system. Chemical exposed for 10 days with the zebrafish. Stock solutions of test chemical were made up with acetone. Appropriate volumes of stock solution were then pipetted into the dishes and evaporated to dryness at room temperature. Control dishes were prepared in each test series by evaporating the greatest volume of acetone used in the corresponding series. The dilution water (50 ml) was added to the dishes. Test animal not fed during the test. But before test Broodstock fish were fed once with dry food (Tetramin) and once with live water fleas each day, except for the weekends when only dry food was given. After each breeding the fishes were returned to the stock aquaria, in order to keep the sexes separated from each other. The eggs were collected approx 5 hrs after fertilization, which corresponds to development stage 13-14. They were rinsed from fecal matters and viable eggs were transferred to Petri-dishes containing toxicant in dilution water. In each petri-dish 10 – 15 viable eggs/dish added. Eggs were considered dead as they turned opaque. Hatched larvae were considered dead when no response to mechanical stimulation occurred. The survival time recorded for each egg or larva is identical to the exposure time when each individual was found dead in connection with the countings. After the exposure of test chemical Methyltrioctylammonium chloride for 10 days with test organism zebrafish effect on the embryos were observed. The LC50 was determine at 0.028 MG/L which was calculated by probit analysis. Thus based on the LC50 value, chemical consider to be toxic and can be consider to be classified as aquatic acute 1/ chronic 1 category as per the CLP classification criteria.

Short term acute toxicity of fish rainbow trout, Salmo guirdneri was evaluated for test material Methyltrioctylammonium chloride in static condition for 120 h. Local tap water was used for the test . At each concentration of test material ten fry were exposed in a volume of 2 dm-3 for at least 120 h. Median lethal concentration (LC50) was calculated by graphic estimation on log-probit paper. The LC50 value were noted at 24,48,96 and 120 h. The medial lethal effect concentration (LC50) was observed to be 0.050 mg/l when observed for 120 h .

On the basis of above effect concentration it can be concluded the test material is highly toxic to fish and hence, can be classified as aquatic chronic 1 as per CLP criteria.

Thus based on the above studies, chemical consider to be toxic and classified as aquatic acute 1/ chronic 1 as per the CLP criteria.

Short term toxicity to invertebrates:

Summarized result from experimental reports and peer reviewed journals for the toxicity of test chemical on the mortality of invertebrates were studied and mention as follows:

In the first experimental study from experimental report 2018, Aim of this study was to assess the short term toxicity of Methyltrioctylammonium chloride to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs. The stock solution 20 mg/l was prepared by dissolving colourless dense liquid in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample in reconstituted water. 0, 0, 0.006, 0.012, 0.025, 0.050, 0.100, 0.200 mg/l concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance Methyltrioctylammonium chloride, in Daphnia magna was determined to be 0.031 mg/L on the basis of mobility inhibition effects in a 48 hour study. Thus EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic acute 1 / chronic 1 category as per the CLP classification criteria.

Above experimental study was supported by the study from peer reviewed journal. Aim of the study was to determine the effect of test chemical on the growth and abnormalities of daphnia magna in the presence and absence of acetone. Stock solutions were made either in acetone or in water. For stock solutions in water appropriate amounts of Aliquat 336 and 5 mL deionized water were sonicated in a 10-mL vial for 30 s, diluted by Standard. Reference Water and allowed to equilibrate for one h. Final test concentrations, which were selected by progressive bisectioning on logaritmic scale, were prepared by diluting appropriate volumes of stock solutions in SRW up to 190 mL, and left for equilibration for 2 h. The final I0 mL SRW was then added together with the animals. Less than 66 hrs old daphnia magna were used in the study. 9-14 animals per concentration was added and performed in duplicates or triplicates. The animals were classified as dead when their second antenna did not move within 15 s after irrigation by a water stream from the handling pipet. Dead and live animals were recorded after 3, 24, 48, 72, and 96 h. After the exposure of test chemical with the water flea daphnia magna, the LC50 was determine to be ranges from 0.0316 – 0.0097 mg/l in the absence of acetone. In the presence of acetone the LC50 was ranges from 0.0129 – 0.0157 mg/l.

Similarly in the third study from peer reviewed journal, the study was conducted to access the level of toxic effect of test chemical Methyltrioctylammonium chloride on the mobility of daphnia magna after the exposure period of 48 hrs. Toxicant solutions (200 ml) were prepared by addition of pre-made Stock solutions of the toxicants to SRW (Standard reference water). Stock solutions were made by dissolving weighed amounts of the toxicants in acetone. stock solution was made for each concentration in order to produce desired nominal concentrations on addition of 50 µl stock solution to 200ml water (toxicant dilution factor 4000 x : final acetone conch 250 µl /I). For the stock culture the < 24 hrs old water fleas were kept in a 501 glass tank filled with 401 aged, aerated tap water, the composition of which has been reported previously. Twice a week 1/4 of the volume was changed and approx. 2/3 of the neonates were removed in order to avoid crowding and stagnation of the culture. 300 ml glass beakers filled with 200 ml of stock solution having headspace of 100 ml were used. Mortality was recorded after 3, 24, 48, 72 and 96 hr. Other reactions of the animals like swimming at the surface and general activity (swimming or resting) were also recorded. After the exposure of test chemical Methyltrioctyl -ammonium chloride with daphnia magna for 96 hrs, the LC50 was determine to be 0.031 mg/l on the basis of mortality of daphnia magna. After 24 hrs the LC50 was determine to be 0.088 MG/L. After 48 and 72 hrs the LC50 was determine at 0.052 MG/L and 0.039 MG/L. based on the LC50 value, chemical consider to be toxic and can be consider to be classified as aquatic acute 1 / chronic 1 as per the CLP classification criteria.

Thus based on the overall studies chemical Methyltrioctylammonium chloride (CAS No. 5137 -55 -3), consider to be toxic and can be consider to be classified as aquatic acute 1 / chronic 1 as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

Based on the various experimental data for the target chemical study have been reviewed to determine toxic nature of Methyltrioctylammonium chloride

(CAS N O. 5137-55-3) on the growth and other activity of algae. The studies are as mentioned below:

The first key study is from experimental report 2018. Aim of this study was to evaluate the nature of chemical test chemical Methyltrioctylammonium chloride when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD guideline 201. The stock solution 100 mg/l was prepared by dissolving colourless dense liquid in OECD growth medium. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Various concentrations 0, 0.05, 0.10, 0.20, 0.40, 0.80 mg/l were used. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. Effect on the growth of algae was determine after an exposure period of 72 hrs. The median effective concentration (EC50) for the test substance Methyltrioctylammonium chloride, in algae was determined to be 0.56 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Thus based on the EC50 value, it indicates that the substance is likely to be hazardous to aquatic algae and can be consider to be classified as aquatic acute1 / chronic 2 category as per the CLP classification criteria.

First study was supported by the second study from experimental data from peer reviewed journal. Aim of this study was to determine the long term effect of test chemical Methyltrioctylammonium chloride on thirteen freshwater algal species grown in 250-µl liquid cultures on plastic microtitration plates. Test conducted for 14 days. Stock solutions of the chemicals in sterile distilled water were freshly prepared under aseptic conditions prior to each experiment. pH was adjusted to 6.5-7.5 using HCl or NaOH. Whenever a cosolvent (ethanol) was used, the final concentration never exceeded 0.2%. 13 algal species were used on which effect were measured was Chlamydomonas dysosmos, Chlorella emersonii, Kirchneriella contorta, Monoraphidium pusillum, Scenedesmus obtusiusculus, Selenastrum capricornutum, Klebsormidium marinum, Raphidonema longiseta, Bumilleriopsis filiformis, Monodus subterraneus, Tribonema aequale, Oscillatoriales LPP species, Synechococcus leopoliensis. Axenic strains of 13 algal species were kept on agar slants. Precultures were grown in an inorganic medium, Z8 modified by the addition of Si (0.16 mM) and vitamins (thiamine, 200 µg/liter; biotin, 1 µg/liter; B12, 1 µg/liter). Cultures were continuously illuminated by Cool White fluorescent tubes (General Electric F96 PG 17CWX Power Groove de Luxe) at an irradiance of 10 ± 1 W/m2 at 400-700 nm. The temperature was 20 ± 1°C. Log phase cells were used as inoculum for the growth experiments. Prior to inoculation of the test cultures the precultures were tested for bacterial contamination by streaking onto agar plates. 250 µl Microtitration plate filled with 100 µl and 150 µl of algal cells were use. The resulting geometric concentration series (factor 0.5) covered 4.2 orders of magnitude (14 concentrations on two plates). Based on the complete algal growth inhibition by the test chemical Methyltrioctylammonium chloride in the exposure period of 14 days, the EC100 was observed at 0.5 mg/l and toxicity values ranges from 0.25 – 1 mg/l. Based on the EC100, chemical consider to be toxic and can be consider to be classified as aquatic chronic 2 as per the CLP classification criteria.

Similarly in the third study the toxicity of test material [4-[[4-(diethylamino)phenyl]phenylmethylene]-2,5-cyclohexadien-1-ylidene]diethylammonium acetate on the growth of Chlorella emersonii (strain 21 1/8h) was evalauted . An artificial medium was used at a 25 % dilution for the growth medium. Air was passed through the cultures at a rate of 10 dm³/ h. The cultures were continuously illuminated with cool white fluorescent tubes, giving a light intensity at the front surface of the culture tubes of 10 W m^{-2 .}Growth was followed by measuring absorbance at 540 nm and growth curves were constructed by plotting the logarithm of the absorbance against time. The inoculum was adjusted to give an initial cell concentration corresponding to a chlorophyll concentration of 0.16 µg cm^-3.The exponential growth rate was determined from the linear part of the growth curves by means of linear regression analysis. The effect of test material was observed for exponential growth curve as well as biomass. The effect concentration (EC50) for Exponential growth rate was observed to be in the range 0.5 - 1.0 mg/l and for biomass EC50 was observed to be 0.1 - 0.5 mg/l. Thus, Based on the above effect concentration it was considered that the test material is toxic to aquatic algae and cyanobacteria and can be classified as aquatic acute 1 as per CLP criteria.

Thus based on the overall studies from experimental report and peer reviewed journal, chemical consider to be toxic and can be consider to be classified as aquatic acute 1/ chronic 2 as per the CLP classification criteria.

Toxicity to microorganism:

1.Bioassay for the inhibition of growth of Cellulomonas sp. was performed for the test material Methyltrioctylammonium chloride.Test organism were inoculated in a synthetic medium prepaired by adding glucose 6 mM; (NH4)2HP04 4 mM; K2HPO4 6 mM; KH2P04 10 mM; KCI 7 mM; EDTA 1 mM; vitamins and trace elements. Inoculums were prepared from 100 cm³cultures grown in 250 cm³Erlenmeyer flasks on a rotary shaker.

Initial cell concentration was recorded to be 10⁷cells cm^-3.The experiments were performed in 100cm³Erlenmeyer flasks equipped with a side tube, making direct measurements possible. The flasks contained 50cm³medium. Incubation was made on a rotary shaker, 120 rev min^-1. Growth was measured turbid metrically at 610 nm.The exponential growth rate was determined from the linear part of the growth curves by means of linear regression analysis.

 

The Effect concentration (EC50) of test material Methyltrioctylammonium chloride on Cellulomonas sp. was observed to be in the range 0.2 – 0.4 mg/l for the exponential growth rate and the effect concentration (EC50) on biomass was observed to be in the range 0.1 – 0.2 mg/l.Based on the above effect concentration test material can be considered toxic to microorganisms.

2.Bioassay for the inhibition of growth of Sporocytophaga myxococcoides was performed for the test material Methyltrioctylammonium chloride.Test organism were inoculated in a synthetic medium prepaired by adding glucose 6 mM; (NH4)2HP04 4 mM; K2HPO4 6 mM; KH2P04 10 mM; KCI 7 mM; EDTA 1 mM; vitamins and trace elements. Inoculums were prepared from 100 cm³cultures grown in 250 cm³Erlenmeyer flasks on a rotary shaker.

Initial cell concentration was recorded to be 10⁷cells cm^-3.The experiments were performed in 100cm³Erlenmeyer flasks equipped with a side tube, making direct measurements possible. The flasks contained 50cm³medium. Incubation was made on a rotary shaker, 120 rev min^-1. Growth was measured turbid metrically at 610 nm.The exponential growth rate was determined from the linear part of the growth curves by means of linear regression analysis.

The Effect concentration (EC50) of test material Methyltrioctylammonium chloride onSporocytophaga myxococcoides was observed to be in the range 0.8 - 0.16 mg/l for the exponential growth rate and the effect concentration (EC50) on biomass was observed to be in the range 0.4 – 0.8 mg/l.Based on the above effect concentration test material can be considered toxic to microorganisms.