o-anisic acid

Endpoint summary

Description of key information

Short term toxicity to fish:

According to aquatic toxicity module i.eACD/LabsI-Lab 2.0; Algorithm Version: v5.0.0.184 the lethal concentration (LC50) of the test substance 2-methoxybenzoic acid was predicted to be 360 mg/l on fish species Fathead minnow (Pimephales promelas). The reliability Index (RI) of the LC50 calculation for 2-methylanthraquinone was 0.57(As per model substance have moderate and high reliability if RI > 0.5). Thus reported RII value indicate it has moderate reliability. The LC50 value indicates that the 2-methoxybenzoic acid considered as non-toxic to fish and thus can be considered to be not classified as per the CLP criteria.

Short term toxicity to aquatic invertebrates:

Short term toxicity to aquatic invertebrates (ABITEC, Determination of the inhibition of the mobility of daphnids.2017) was performed in Daphnia magna following the OECD guideline 202. The test was performed for 48 hrs at different concentrations as control 0, 30, 45, 67.5, 100 and 150 mg/L at 95 % confidence limit of 74.5-97.0.Reference substance used was K2Cr207. 5 pieces of daphnids after breeding were used in 4 numbers of replicates of test concentration. The test was performed in temperature 20°C±1°C with taking 25ml per replicate in 50ml glass vessel for 48 hrs. Calculations of EC50 were calculated using nonlinear regression by the software Prism4.0 (Graph Pad Software, Inc., San Diego CA).

After experiment, the EC50 value for short term toxicity to aquatic invertebrates using OECD guideline 202 was determined to be 85.1 mg/l. Based on the value the chemical was considered as classified but as it was readily biodegradable in nature, hence can be considered as not classified as per the CLP regulations.

Toxicity to aquatic algae and cyanobacteria:

Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus using OECD Guideline 201. The stock solution (100 mg/L) was prepared by dissolving white powder in OECD growth medium. The solution was kept in ultrasonic bath for 20 mins. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Conc. of test chemical used for the study was 0, 35, 46, 59, 77 and 100 mg/L, respectively. The test was performed under static conditions in a static fresh water system at a temperature of 23±2°C. Initial cell density of test organism used was 5x10(3) cells/ml. Determination of cell counting involve the use of microscope with counting chamber Cyrus I or electronic particle counter. ErC50 was calculated using non-linear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance 2-methoxybenzoic acid (2-MBA) in Desmodesmus subspicatus was determined to be 78.8 mg/L on the basis of effects on growth rate in a 72 hour study.Based on the value the2-methoxybenzoic acid (2-MBA)was considered as toxic to aquatic organism but as it was readily biodegradable in nature, hence can be considered as not classified as per the CLP regulations.

Additional information

Short term toxicity to fish:

Short term toxicity to fish was summarized with 2 predicted data are as follows:

According to aquatic toxicity module i.eACD/LabsI-Lab 2.0; Algorithm Version: v5.0.0.184 the lethal concentration (LC50) of the test substance 2-methoxybenzoic acid was predicted to be 360 mg/l on fish species Fathead minnow (Pimephales promelas). The reliability Index (RI) of the LC50 calculation for 2-methylanthraquinone was 0.57(As per model substance have moderate and high reliability if RI > 0.5). Thus reported RII value indicate it has moderate reliability. The LC50 value indicates that the 2-methoxybenzoic acid considered as non-toxic to fish and thus can be considered to be not classified as per the CLP criteria.

Using the EPI Suite ECOSAR version 1.11, the short term toxicity on fish was predicted for test substance  2-methoxybenzoic acid (CAS: 579-75-9) on the basis of effects observed in a static freshwater system. The lethal concentration LC50 value for the substance was estimated to be 2783.679 mg/l for fish for 96 hr duration. Based on this value it can be concluded that the 2-methoxybenzoic acid can be considered to be non-toxic to aquatic environment and can be considered as not classified as per the CLP classification criteria.

Based on the predicted data from ACD labs and ECOSAR version 1.11, the toxicity to fish was predicted and based on the values, 2-methoxybenzoic acid (CAS: 579-75-9) was considered to be non-toxic to fish and can be considered to be not classified as per the CLP regulations.

Short term toxicity to aquatic invertebrates:

Toxicity to aquatic invertebrates for 2-methoxybenzoic acid (579-75-9) was summaries with target data and by using functional read across studies are summaries as follows:

Short term toxicity to aquatic invertebrates (ABITEC, Determination of the inhibition of the mobility of daphnids.2017) was performed in Daphnia magna following the OECD guideline 202. The test was performed for 48 hrs at different concentrations as control 0, 30, 45, 67.5, 100 and 150 mg/L at 95 % confidence limit of 74.5-97.0.Reference substance used was K2Cr207. 5 pieces of daphnids after breeding were used in 4 numbers of replicates of test concentration. The test was performed in temperature 20°C±1°C with taking 25ml per replicate in 50ml glass vessel for 48 hrs. Calculations of EC50 were calculated using nonlinear regression by the software Prism4.0 (Graph Pad Software, Inc., San Diego CA).

After experiment, the EC50 value for short term toxicity to aquatic invertebrates using OECD guideline 202 was determined to be 85.1 mg/l. Based on the value the chemical was considered as classified but as it was readily biodegradable in nature, hence can be considered as not classified as per the CLP regulations.

Daphnia tests were conducted by Michael Cleuvers, (2004) on read across 2-acetoxybenzoic acid (50-78-2) following the European Guideline (Commission of the European Communities, 1992) using the water flea. Daphnids were bred in ADaM, a culture medium imitating natural freshwater. Test was performed in 20 degree C with photoperiods of 16 h light/8 h dark. Young daphnids were used in 50 ml of culture volume, each treatment was subdivided in four replicates, each containing 5 daphnia and observed for 24 and 48 hrs. After the experiment the EC50 value for short term toxicity to aquatic invertebrates for 2-acetoxybenzoic acid (50-78-2) was determined to be 88.1 mg/l. Based on the values, 2-acetoxybenzoic acid was considered to toxic to aquatic invertebrates but as it is readily biodegradable in nature hence, 2-acetoxybenzoic acid (50-78-2) was considered to be not classified as per the CLP regulations.

Another study for read across 4-methoxybenzoic acid (100-09-4) from Chemosphere 59 (2005) 255–261, short term toxicity to aquatic invertebrates was performed in Daphnia magna for 48 hrs. The young daphnids were obtained from continuous cultures in 1 liter glass beakers at 21 ± 0.3 degree C, in dechlorinated and conditioned tap water, the medium was renewed in three weeks and daphnids were fed daily with the green alga Selenastrum capricornutum. The pH of test solution was adjusted to 7.45 ± 0.05 with HCL and NaOH. The concentrations of the resultant solutions were checked before and after (48 h) exposure using a UV–visible spectrophotometer. As the experiment starts neonate were placed in 50 ml of beaker with 40 ml of test solution. All experiment was made in four replicates and performed at 21 ± 0.3 degree C under 16 h light: 8 h dark photoperiod. Immobility was observed after 24 and 48 hrs and EC 50 value was calculated. After the experiment the EC 50 value of 4-methoxybenzoic acid (100-09-4) for Daphnia magna was determined to be 943 mg/l in 48 hrs. Based on the CLP regulation the 4-methoxybenzoic acid is considered to be not toxic to aquatic invertebrates and can be concluded to be not classified as per the CLP regulations.

From the same reference, another read across Benzoic acid (65-85-0) was also studied and the EC 50 value of Benzoic acid (65-85-0) for Daphnia magna was determined to be 860 mg/l in 48 hrs.

Based on the results of target and result of read across chemical, it was considered that 2-methoxybenzoic acid (579-75-9) was likely to be non-toxic to aquatic invertebrates and can be considered to be not classified as per the CLP regulations.

Toxicity to aquatic algae and cyanobacteria:

Toxicity to algae for 2-methoxybenzoic acid (579-75-9) was summarised with data for target and by using functional read across studies was summaries as follows:

Freshwater algal growth inhibition test (Abitec , Freshwater algal growth inhibition test, report number 185/17,2017) was carried out on Desmodesmus subspicatus using OECD Guideline 201. The stock solution (100 mg/L) was prepared by dissolving white powder in OECD growth medium. The solution was kept in ultrasonic bath for 20 mins. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Concentration of test chemical used for the study were 0, 35, 46, 59, 77 and 100 mg/L, respectively. The test was performed under static conditions in a static fresh water system at a temperature of 23±2°C. Initial cell density of test organism used was 5x10(3) cells/ml. Determination of cell counting involve the use of microscope with counting chamber Cyrus I or electronic particle counter. ErC50 was calculated using non-linear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance 2-methoxybenzoic acid (2-MBA) in Desmodesmus subspicatus was determined to be 78.8 mg/L on the basis of effects on growth rate in a 72 hour study.Based on the value the 2-methoxybenzoic acid (2-MBA)was considered as toxic to aquatic organism but as it was readily biodegradable in nature, hence can be considered as not classified as per the CLP regulations.

Study for read across Benzoic acid from Environmental Pollution (Series A), Vol. 29, Pg. no. 71-80, 1982 was carried out using cyanobacteria (blue-green algae) Anabaena inaequalis, A. cylindrica,and A.variabilis, and the green algae Chlorella pyrenoidosa and Scenedesmus quadricauda as test organisms. The study was based on the effects of the test compound Benzoic acid on algae in a static fresh water system. Stock solutions of test chemical were prepared in pesticide grade acetone.The proper level of acetone (0.1 %v/v) to use in each bioassay system was determined using the solvent-pesticide interaction analysis technique.Chemical was tested at a minimum of five concentrations ranging from 0 to 100 mg/l (nominal concentration). Test organisms used for the study include (blue-green algae) Anabaena inaequalis, A. cylindrica, and A.variabilis, and the green algae Chlorella pyrenoidosa and Scenedesmus quadricauda, respectively.

Cultures of Anabaena inaequalis and A. cylindrica were maintained in a liquid nitrogen-free medium at a temperature of 20°C and a light intensity of 7000 lux on a 12 h light-dark cycle whereas the other cultures such as A.variabilis, Chlorella pyrenoidosa and Scenedesmus quadricauda were maintained under the same conditions except the growth medium was supplemented with 1.5 g NaNO3/l. Photosynthesis was assayed by following the uptake of 14CO2 from NaH14CO3. Plastic tissue culture flasks with a total internal volume of 74 ml were employed as assay chambers. Each contained 9.9ml of cell suspension (containing 6-5 × 104cyanobacterial or 1.0 × 105green algal cells/ml), 0.1 ml of radioisotope (to give a final activity of 0.1µCi/ml) and 0.01 ml of test chemical. The flasks were incubated for 3 h and photosynthetic activity was assayed. Per cent inhibition values were calculated relative to photosynthetic activity in control systems (solvent only) and EC50 values determined by probit or regression analysis, where applicable. Analyses for significant differences (p = 0.05) were performed using Dunnett's testand Duncan's multiple range test.

Based on the effect of Benzoic acid on the photosynthesis of the test organisms, the EC50 value was determined to be 5, 60, 55, 60 and 75 mg/l for Anabaena inaequalis, A. cylindrical, A.variabilis, Chlorella pyrenoidosa and Scenedesmus quadricauda respectively. Thus, based on this value, it can be concluded that the test chemical Benzoic acid can be considered as toxic to aquatic organisms. Since the chemical is readily biodegradable in nature, Benzoic acid can be considered as non-toxic to aquatic organisms and thus cannot be classified as hazardous as per the CLP criteria.

Short term toxicity to Pseudokirchneriella subcapitata (green algae) study was carried out on same read across benzoic acid for 48 hrs from Ecotox database, 2016 .The study was based on the effects of the test compound on Pseudokirchneriella subcapitata in a static fresh water system. On the basis of population of the test organism Pseudokirchneriella subcapitata (green algae), the 48 hr EC50 value was determined to be 207.5 mg/l.

Based on the results of target and result of read across chemical, it was considered that 2-methoxybenzoic acid (579-75-9) was likely to be non-toxic to aquatic algae and can be considered to be not classified as per the CLP regulations.