Methyl anthranilate

Endpoint summary

Description of key information

Short-term toxicity to fish:

A study was conducted to determine the toxic effects of test chemical on fish. The test organisms used in this study were Atlantic salmon (Salmo safar L.) and Rainbow trout (Oncorhynus mykiss Richardson). Nominal concentrations prepared for Atlantic salmon and Rainbow trout were 1, 6, 13, 25, and 50 mg/L and 0, 1, 5, 10, 15, 20, 25 and 50mg/L,. Fishes were maintained in holding tanks for two weeks prior to testing on a 12-h daylight photoperiod. Dilution water which was derived from well water was run through a series of filters. Temperature was monitored and recorded every 30 min. After the exposure of test chemical, both the species showed loss of equilibrium at 50 mg/L within 25–30 s, eventually immobility was observed on the bottom of the vessel. Hence, after 96h of exposure, the LC50 value for Atlantic salmon and Rainbow trout was determined to be 32.35 mg/L and 22.91 mg/L, respectively.

However, for Atlantic salmon LC50 value at 24h, 48h and 72h was 34.28 mg/L, 33.31 mg/L and 32.35 mg/L, respectively with NOEL at 6 mg/L. Whereas for Rainbow trout LC50 value at 24h, 48h and 72h was 23.47 mg/L, 23.19 mg/L and 23.19 mg/L with NOEL at 5 mg/L.

Since the test chemical is readily biodegradable the test chemical is considered to be nontoxic to fish and cannot be classified according to CLP regulation.

Long-term toxicity to fish:

Based on the prediction done by EPI suite, ECOSAR version 1.11, on the basis of similarity of structure to chemicals for which the aquatic toxicity has been previously measured by structure-activity relationships (SARs) program, the LC 50 value for long term toxicity to aquatic invertebrates was predicted.

On the basis of this program, the LC 50 value for long term toxicity to fish was predicted to be 7.22 mg/l for test chemical in 28 days. Based on this value it can be concluded that the substance is non- toxic to aquatic environment and cannot be classified as per the criteria mentioned in CLP regulation.

 

Short-term toxicity to aquatic invertebrates:

An experimental study was conducted to determine the short term toxicity of test chemical to aquatic invertebrates. The test organism used in this study was Daphnia magna less than 24h old. The experiment was conducted in the static system using freshwater. After the exposure of test chemical for 48 h, the effect concentration EC50 and NOEL was determined to be 1 8.2 mg/L and 5.5mg/L, respectively. Since the test chemical is readily biodegradable the test chemical is considered to be nontoxic to Fish and cannot be classified according to CLP regulation.

Long-term toxicity to aquatic invertebrates:

Based on the prediction done by EPI suite, ECOSAR version 1.11, on the basis of similarity of structure to chemicals for which the aquatic toxicity has been previously measured by structure-activity relationships (SARs) program, the EC 50 value for long term toxicity to aquatic invertebrates was predicted.

On the basis of this program, the EC 50 value for long term toxicity to aquatic invertebrates was predicted to be 4.36 mg/l for test chemical in 21 days. Based on this value it can be concluded that the substance was considered to be nontoxic to aquatic environment and cannot be classified as per the criteria mentioned in CLP regulation.

 

Toxicity to aquatic algae and cyanobacteria:

An experiment was conducted to determine the short term toxicity of test chemical to aquatic alagae. The test organism used in this study were Oscillatoria perornata and Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum. Test chemical solution was prepared by using Methanol was used as a solvent. The experiment was conducted in static system and test chemical was exposed to algae for 5 days. The 96-well microplates were used which were placed in a growth chamber at 28-30°C. Hence, after 5 days of exposure the LOEC value of test chemical was determined to be 151.16 mg/L on the basis of this the EC50 value was determined >151.16 mg/L. By considering this EC50 value it is concluded that test chemical is nontoxic to aquatic algae and cannot be classified according to CLP regulation.

 

Toxicity to microorganisms:

A study was conducted to determine the minimum inhibition concentration on microorganisms for 24h. Organisms used for the study were Staphylococcus aureus, Escherichia coli, Candida albicans and diphtheroid, which were obtained from Monmouth Medical Center, Long Branch, New Jersey and the Department of Dermatology, University of Pennsylvania, respectively. Nominal concentrations used were 0.1, 1.0, 10, 100, and 1000 mg/L. MIC was determined by Liquid Culture method in which TGY medium was used without the agar, while the pH was 7.0. Thus, The Minimum Inhibitory Concentration of the test chemical for 24 h on Staphylococcus aureus, Escherichia coli and Candida albicans, diphtheroid was determined to be >1000mg/L and 1000 mg/L, respectively.

Additional information

Short-term toxicity to fish:

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

The first study was reviewed from the authoritative databases and peer reviewed journal in this study an experiment was conducted to determine the toxic effects of test chemical on fish. The test organisms used in this study were Atlantic salmon (Salmo safar L.) and Rainbow trout (Oncorhynus mykiss Richardson). Nominal concentrations prepared for Atlantic salmon and Rainbow trout were 1, 6, 13, 25, and 50 mg/L and 0, 1, 5, 10, 15, 20, 25 and 50mg/L,. Fishes were maintained in holding tanks for two weeks prior to testing on a 12-h daylight photoperiod. Dilution water which was derived from well water was run through a series of filters. Temperature was monitored and recorded every 30 min. After the exposure of test chemical, both the species showed loss of equilibrium at 50 mg/L within 25–30 s, eventually immobility was observed on the bottom of the vessel. Hence, after 96h of exposure, the LC50 value for Atlantic salmon and Rainbow trout was determined to be 32.35 mg/L and 22.91 mg/L, respectively. However, for Atlantic salmon LC50 value at 24h, 48h and 72h was 34.28 mg/L, 33.31 mg/L and 32.35 mg/L, respectively with NOEL at 6 mg/L. Whereas for Rainbow trout LC50 value at 24h, 48h and 72h was 23.47 mg/L, 23.19 mg/L and 23.19 mg/L with NOEL at 5 mg/L.

Next study was also reviewed from journal and authoritative databases in this astudy was conducted to determine the toxic effects of test chemical on fish. The test organisms used in this study were Bluegill (Lepomis macrochirus Raf.) and Channel catfish (Ictulurus punctatus Raf.). Nominal concentrations prepared for both the species were 5, 10, 20, 40, 50 and 100 mg/L. Fishes were maintained in holding tanks for two weeks prior to testing on a 12-h daylight photoperiod. Dilution water which was derived from well water was run through a series of filters. Temperature was monitored and recorded every 30 min. After the exposure of test chemical, both the species indicated abnormal behaviour, eventually mortality was observed. The LC 50 value was determined by using probit analysis method and after 96 h of exposure of test chemical to fish , the LC50 value for Bluegill and Catfish was determined to be 9.12 mg/L (95% C.I. = 7.98 - 10.51 mg/L) and 16.23 mg/L (95% C.I = 11.57 - 22.47 mg/L), respectively. However, for Bluegill LC50 value at 24h, 48h and 72h was 19.80 mg/L, 9.12 mg/L and 9.12 mg/L, respectively with NOEL at 7 mg/L. Whereas for Catfish LC50 value at 24h, 48h and 72h was 20.08 mg/L, 17.35 mg/L and 16.94 mg/L with NOEL at 7 mg/L.

In addition to above next study was reviewed from authoritative database in this a study was conducted to determine the short-term toxicity of test chemical to fish. The test organisms used in this study was Oncorhynchus mykiss (previous name: Salmo gairdneri). The test chemical was exposed to Rainbow trout for 96h. The experiment was conducted in flow through system using freshwater. After the exposure of test chemical, the LC50 and NOEL was determined to be 25.4 mg/L and 7.36 mg/L, respectively.

Another study was also reviewed from authoritative database in this astudy was conducted to determine the short-term toxicity of test chemical to fish. The test organisms used in this study was Lepomis macrochirus. The test chemical was exposed to Lepomis macrochirus for 96h. The experiment was conducted in flow through system using freshwater. After the exposure of test chemical, the LC50 and NOEL was determined to be 42.56 mg/L and 33.6 mg/L, respectively.

 

Next study was reviewed from journal in this a study was conducted to determine the short-term toxicity of test chemical to fish. The test organisms used in this study was Ictalurus punctatus (common name: Channel catfish). Fishes of length 9.9 - 21cm were stocked in the holding pen pond. Test vessel used for the study was 6,664-L holding tank which was 3 m in diameter. Test chemical was applied to the surface at one of the following application rates: 0, 2.2, 20, 40, 100, or 200 kg/ha. The formulation (test chemical) was seen as an oily sheen covering the water's surface. The analytical monitoring of test chemical was done and for that water samples were taken at depths of 0.03 and 0.35 m from the surface to quantify the amount of test chemical. During the 12-h exposure period Ictalurus punctatus behaved normally and kept mostly to the bottom of the tank at a depth of 0.9 m. After the exposure of test chemical for 12 h the LD50 value of test chemical on test organism was determined to be 20 mg/L.

The last study was conducted to determine the short-term toxicity of test chemical on Cyprinus carpio for 68 h. The test fishes were captured with an alternating-current electric boat shocker in the New York State Barge Canal. The test chemical was exposed to fish by oral route and it was force feed in the form of gelatin capsules. Nominal concentrations used were 184, 191 and 204 mg/Kg. After the exposure of test chemical effect were observed. If a fish acted or looked other than normal it was considered to be sick. If no movement occurred it was recorded as dead. However, mortality was not observed at the concentration range from 184 - 204 mg/Kg. Thus, from the above observation, it can be concluded that the NOEC value of the test chemical on Cyprinus carpio after 68 h was determined to be in the range 184-204 mg/Kg.

By considering results of all the studies mentioned above the LC 50 value of test chemical was determined to be in range from 9.12 mg/L to 42.56 mg/L. Since the test chemical is readily biodegradable the test chemical is considered to be nontoxic to Fish and cannot be classified according to CLP regulation.

Long-term toxicity to fish:

Predicted study for target chemical and experimental study for its structurally similar read across chemical have been reviewed to determine the long term toxicity of test chemical to fish and their results are summarized below.

The first study was based on the prediction done by EPI suite, ECOSAR version 1.11, on the basis of similarity of structure to chemicals for which the aquatic toxicity has been previously measured by structure-activity relationships (SARs) program, the LC 50 value for long term toxicity to aquatic invertebrates was predicted. On the basis of this program, the LC 50 value for long term toxicity to fish was predicted to be 7.22 mg/l for test chemical in 28 days. Based on this value it can be concluded that the substance is non- toxic to aquatic environment and cannot be classified as per the criteria mentioned in CLP regulation.

Next study was reviewed from Arch. Environ. Contain. Toxicol. in this an experiment was conducted to determine the long term toxicity of test chemical to fish by using 28-day Larval Test. In this test organism used was Oryzias latipes (Japanese Medaka) and it was fed Brine shrimp twice a day during the experiment. The Stock solutions of test chemical were prepared by dissolving test chemical in Lake Superior water, using a high speed stirrer. Stock solutions were transferred to a glass stock bottle inside the vented diluter enclosure using Teflon tubing and air pressure. During each test, a predetermined volume (ml/min) of stock solution was continuously pumped from the stock bottle into the mixing cell of the diluter system. Sampling was done twice weekly and alternately from each complete set of replicate tanks (6 tanks). The analytical monitoring was done by using A Hewlett Packard 5730 A gas chromatograph (GC) equipped with a flame ionization detector (FID) linked to an HP 3350 lab automation system. The experiment was conducted in Continuous-flow mini-diluter exposure systems in Glass aquaria of size 18.5 × 14.0 × 13.0 cm deep. The number of organisms per vessel were 20 and photoperiod of 16 h was given to test organisms. The measured concentration of test chemical were determined to be 0.27 ± 0.07, 0.47 ± 0.16, 0.92 ± 0.33, 1.92 ± 0.45 and 4.85 ± 0.82 mg/Land it control tank it was determined to be <0.25 mg/L. The statistical method used to calculate maximum acceptable test chemical(toxicant) concentrations (MATCs) was one-way analysis of variance with an F-test of significance (p = 0.05) Growth data and normalized hatch and survival data and to subsequent analysis with Dunnett's one-sided comparison (p = 0.05) was used. After exposure of test chemical for 28 days to Oryzias latipes the MATC for test lies between 0.92 and 1.92 mg/L and chronic value was determined to be 1.33 mg/L.

Last study was reviewed from Aquatic Toxicology (1990) in this an experiment was conducted to determine the long term toxicity of test chemical to fish determined by using early life stage (ELS) in which fish are exposed during embryogenesis and larval development. The test organism used in this study was Danio rerio (previous name: Brachydanio rerio).Fertilized eggs of zebra fish (Brachydanio rerio) in the blastula stage were obtained from a stock culture at the TNO laboratory of 50-100 eggs (<6 h after spawning) 40 eggs per concentration were used Upon completion of hatching (4-5 days), the fry were transferred into two vessels per concentration of test chemical. The test solution was prepared by using vehicle DMSO (dimethyl suiphoxide) but the DMSO concentrations were kept below 100 µl/L the ratio between the concentrations was 1.8.DMSO were verified in solvent control experiments. The experiment was conducted in semi-static condition using reconstituted water which was prepared from groundwater obtained from a locality near Linschoten, to which several salts were added at 24 + 2°C temperature. The Photoperiod given was of 12 h. The test solutions were renewed 3 times a week. The analytical monitoring of test chemical was done before and after renewal of the test solutions by using Waters 710B HPLC equipped with a Waters 6000A pump and a Kratos Spectro324 flow UV detector at 220/240 nm and a Guard 30-40/zm precolumn (Chrompack) and a Vydac 201 TPB 5 pm 100 mm × 3 mm column (Chrompack) were used. After 28 days of exposure of test chemical to fish the Differences in mean survival in the experimental concentrations were tested against blank control by means of a χ2 test and the lethal concentration at which 50% mortality (LC50) was observed and NOEC value was determined to be 39 mg/L( 95% confidence (30-51) )and 1.8 mg/L respectively.

By considering results of all the study mentioned above the LC50 value of test chemical was determined to be in range of 1.33 to 39 mg/L after exposure of test chemical for 28 days to fish. By considering range EC50 value it is concluded that test chemical non-toxic to aquatic organisms and cannot be classified as per CLP regulation.

 

Short-term toxicity to aquatic invertebrates:

Various experimental studies have been reviewed from different sources for target chemical and its structurally similar read across chemical to determine the short term toxicity of test chemical to aquatic invertebrates and their results are summarized below.

The first study was reviewed from authoritative database in this study an experimental study was conducted to determine the short term toxicity of test chemical to aquatic invertebrates. The test organism used in this study was Daphnia magna less than 24h old. The experiment was conducted in the static system using freshwater. After the exposure of test chemical for 48 h, the effect concentration EC50 and NOEL was determined to be 18.2 mg/L and 5.5mg/L, respectively. Since the test chemical is readily biodegradable the test chemical is considered to be nontoxic to aquatic invertebrates and cannot be classified according to CLP regulation.

Next study was reviewed from the study report in this 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). The test organism used in this study was Daphnia magna less than 24h old. The stock solution of test chemical was prepared by dissolving test chemical in acetone at concentration of 100 mg/l. Test solutions of required concentrations were prepared by mixing the stock solution of the test chemical in reconstituted water. The nominal concentration used in the study were 10, 20, 30, 45, 100 mg/l. The experiment was conducted in static system and test chemical was exposed to Daphnia magna. 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 chemical, in Daphnia magna was determined to be 43.2 mg/L on the basis of mobility inhibition effects in a 48 hour study.

 

 

Last study was reviewed from secondary source in this study an experiment was conducted to determine the short term toxicity of test chemical to aquatic invertebrates. The test organism used in this study was Daphnia magna. The experiment was conducted in the static system using freshwater. After the exposure of test chemical for 24 h, the effect concentration EC50 was determined to be 50 mg/L. Since the test chemical is readily biodegradable the test chemical is considered to be nontoxic to aquatic invertebrates and cannot be classified according to CLP regulation.

By considering results of all the studies mentioned above the EC 50 value of test chemical was determined to be in range from 18.2 mg/L to 50 mg/L. Since the test chemical is readily biodegradable the test chemical is considered to be nontoxic to aquatic invertebrates and cannot be classified according to CLP regulation.

 

Long-term toxicity to aquatic invertebrates:

Based on the prediction done by EPI suite, ECOSAR version 1.11, on the basis of similarity of structure to chemicals for which the aquatic toxicity has been previously measured by structure-activity relationships (SARs) program, the EC 50 value for long term toxicity to aquatic invertebrates was predicted.

On the basis of this program, the EC 50 value for long term toxicity to aquatic invertebrates was predicted to be 4.36mg/l for test chemical in 21 days. Based on this value it can be concluded that the substance was considered to be non-toxic to aquatic environment and can be classified ''not classified'' as per the criteria mentioned in CLP regulation.

 

Toxicity to aquatic algae and cyanobacteria:

Various experimental studies have been reviewed from different sources for target chemical and its structurally similar read across chemical to determine the short term toxicity of test chemical to aquatic algae and their results are summarized below.

The first experimental study was reviewed from journal Bull. Environ. Contam. Toxicol. (2001)in this an experiment was conducted to determine the short term toxicity of test chemical to aquatic alagae. The test organism used in this study were Oscillatoria perornata and Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum. Test chemical solution was prepared by using Methanol was used as a solvent. The experiment was conducted in static system and test chemical was exposed to algae for 5 days. The 96-well microplates were used which were placed in a growth chamber at 28-30°C. Hence, after 5 days of exposure the LOEC value of test chemical was determined to be 151.16 mg/L on the basis of this the EC50 value was determined to be >151.16 mg/L. By considering this EC50 value it is concluded that test chemical is nontoxic to aquatic algae and cannot be classified according to CLP regulation.

 

In next study an experiment was conducted to determine the toxicity of test chemical to aquatic algae according to OECD Guideline 201 (Alga, Growth Inhibition Test). The test organism used in this study was Desmodesmus subspicatus (previous name: Scenedesmus subspicatus).The stock solution was prepared by dissolving test chemical in acetone of 200 mg/L. Test solutions of required concentrations were prepared by mixing the stock solution of the test chemical with OECD growth medium inoculum culture. The nominal concentrations of test chemical used in the study were 3.8, 4.7, 9.0, 11.3, 90.1 mg/l. 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 chemical, in algae was determined to be 111.7 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Based on the EC50 value, which indicates that the substance is likely to be non-hazardous to aquatic algae and cannot be classified as per the CLP classification criteria.

 

 

In last study an experiment was conducted to determine the toxicity of test chemical to aquatic algae according to OECD Guideline 201 (Alga, Growth Inhibition Test). The test organism used in this study was Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). The experiment was conducted in static system using freshwater. Based on the effect on growth rate of the test organism and after exposure of test chemical to aquatic algae the EC50 value was determined to be 110 mg/l. By considering EC50 value it is concluded that the test chemical is non-toxic to aquatic organisms and cannot be classified as hazardous as per the CLP criteria.

By considering results of all the studies mentioned above the EC 50 value of test chemical was determined to be in range from 110 mg/L to >151.16 mg/L. By considering this range EC50 value it is concluded that the test chemical is non-toxic to aquatic organisms and cannot be classified as hazardous as per the CLP criteria.

 

Toxicity to microorganisms:

Based on the experimental data for the target as well as read-across analogues which are extracted by using mechanistic approach and functionally and structurally similar to the target chemical toxicity of test chemical was determined on the basis of growth inhibition of microorganisms. The studies are summarized as below:

 

A study was conducted to determine the minimum inhibition concentration on microorganisms for 24h. Organisms used for the study were Staphylococcus aureus, Escherichia coli, Candida albicans and diphtheroid, which were obtained from Monmouth Medical Center, Long Branch, New Jersey and the Department of Dermatology, University of Pennsylvania, respectively. Nominal concentrations used were 0.1, 1.0, 10, 100, and 1000 mg/L. MIC was determined by Liquid Culture method in which TGY medium was used without the agar, while the pH was 7.0. Thus, The Minimum Inhibitory Concentration of the test chemical for 24 h on Staphylococcus aureus, Escherichia coli and Candida albicans, diphtheroid was determined to be >1000mg/L and 1000mg/L, respectively. 

 

A study was conducted to determine the toxicity of test chemical on microorganisms for 24h. Organisms used for the study were Staphylococcus aureus, Escherichia coli, Candida albicans and diphtheroid, which were obtained from Monmouth Medical Center, Long Branch, New Jersey and the Department of Dermatology, University of Pennsylvania, respectively. Petri Plate-Paper Disc method was used, wherein, TGY agar was used. The Paper discs of were soaked with 20µl of the 10% test solutions. Thus, the zone diameter of the test chemical for 24 h on Escherichia coli, Staphylococcus aureus, Candida albicans and diphtheroid by Petri Plate-Paper Disc method was determined to be 14mm, 12mm, 16mm and 0mm respectively.

 

A study was conducted to determine the toxicity of test chemical on Pseudomonas putida for 16h. Study was performed according to ISO 10712 (Water quality – Pseudomonas putida growth inhibition test (Pseudomonas cell multiplication inhibition test. After the exposure of test chemical for 16 h, the EC50 and EC10 value were determined to be 380mg/L and 140mg/L, respectively