Sodium salicylate

Endpoint summary

Description of key information

Short-term toxicity to fish

Study was conducted to access the effect of test chemical on the growth of zebra fishDanio rerio. The experiment was performed according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring.

Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fish. A static procedure was used to determine the lethal concentration of test chemical for fish. The zebra fish were exposed to the concentration of 100mg/ L, and test was conducted for the period of 96 hrs. the exposed fish were observed for any visual behavioural abnormality and mortality.

After the 96 hours of exposure to test chemical at nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

Long-term toxicity to fish

Based on the prediction done using theEPI Suite ECOSAR version 1.11, the long term toxicity on fish was predicted for test chemical. On the basis of effects observed in a static freshwater system, the effect concentration NOEC value for the substance is estimated to be 6.867 mg/l for species for 28 days duration. Based on this value, and the test chemical is readily biodegradable it can be concluded that the test substance can be considered as non-toxic  to fish at environmentally relevant concentrations and can be considered as "not classified"  as per the CLP classification criteria.

Short-term toxicity to aquatic invertebrates

Short term toxicty test was carried out according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) with Daphnia magna as a test organisms.

The effective concentration (EC10) value of test chemical in Daphnia magna in a 24 hr study on mortality effect was determined to be 304 mg/L. Thus,considering the CLP Criteria for aquatic classification of the substance , it is concluded that test substance does not exhibit short term toxicity to aquatic invertebrate (Daphnia magna).

 

Long-term toxicity to aquatic invertebrates

Based on the prediction done using the EPI Suite ECOSAR version 1.11, the long term toxicity on daphnia was predicted for test chemical. On the basis of effects observed in a static freshwater system, the effect concentration NOEC value for the substance is estimated to be 4.126 mg/l for daphnia species for 21 days duration. Based on this value, and the test chemical is readily biodegradable it can be concluded that the test substance is considered as non-toxic  to daphnia magna at environmentally relevant concentrations and can be considered as "not classified"  as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria

Experimental study performed for the target chemical suggests thatThis study was designed to assess the effect of the test substance on the growth of green algaChlorella vulgaris. The study was conducted in accordance with “OECD guideline for testing of chemicals No. 201 – Alga growth inhibition test”.

The effect of test substance was studied on the growth of fresh water green algaChlorella vulgaris.The study was conducted following OECD guideline 201- Alga growth inhibition test. The test concentration chosen for the study were 6.25 mg/l, 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l and 200 mg/l were prepared using stock solution of the test substance using de-ionized water. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of test substance. EC50 calculated from probit analysis was observed to be 48.29 mg/L and EC50 graphically was observed to be 63.10 mg/L. As the test chemical is readily biodegradable, it cannot be classfied as per CLP classification category

Toxicity to microorganisms

The toxicity of test chemical to microorganisms determined to be in range of 200 to > 1000 mg/l.

Additional information

Short-term toxicity to fish

Following studies of target chemical includes experimental data to conclude the toxicity extent of test chemical towards fish is summarized as follows:

Study was conducted to access the effect of test chemical on the growth of zebra fishDanio rerio. The experiment was performed according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring.

Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fish. A static procedure was used to determine the lethal concentration of test chemical for fish. The zebra fish were exposed to the concentration of 100mg/ L, and test was conducted for the period of 96 hrs. the exposed fish were observed for any visual behavioural abnormality and mortality.

After the 96 hours of exposure to test chemical at nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

The above study is supported by another experimental study from the handbook Acute Tox. of Organic Chemicals to Fathead Minnows (Pimephales promelas) 1985, suggests that the Acute Toxic studies of the test chemical was studied on the fresh water fish Pimephales promelas. The test was performed for the period of 96 hrs, based on the mortality of the fishes, the median lethal concentration was determined. Stocks were prepared in 18.9 L stainless steel carbonation vessels, by mixing test substance in the Lake Superior water. The median lethal concentration of test chemical (LC50) was determined to be 1370 mg/L. Most of the effected fishes at concnentration greater than 1370 have lost their schooling behaviour and were swimming at the surface.

Further it is supported by similar experimental study from the handbook Acute Tox. of Organic Chemicals to Fathead Minnows(Pimephales promelas) 1985. Test chemical was evaluated for the determination of median lethal concentration (LC50) using fish as a animal model, for the tenure of 96 h. Test chemical was exposed to the nominal concentrations, ranging from 0 to 3080 with the geometric mean of 1.53. The experiment was conducted in the flow through condition.

The basis for the determination of the LC50 concentration was mortality. Most of the fishes lost their schooling behaviour, equilibrium and were hyperactive prior to their death. The median lethal concentration of the fish was determined as 2160 mg/L.

Thus based on the effect concentrations which is in the range 100 mg/L to 2160 mg/lgive the conclusion that test substance is likely to be not toxic to aquatic environment at environmentally relevant concentrations and applying weight of evidence approach it can be considered to be “not classified” as per the CLP classification criteria.

Long-term toxicity to fish

Based on the prediction done using theEPI Suite ECOSAR version 1.11, the short term toxicity of test chemical on green algae was predicted. On the basis of effects observed in a static freshwater system, the effect concentration NOEC value for the substance is estimated to be 6.867 mg/l for fish species for 28 days duration. Based on this value, and the test chemical is readily biodegradable it can be concluded that the test substance can be considered as non-toxic  to fish at environmentally relevant concentrations and can be considered as "not classified"  as per the CLP classification criteria.

Short-term toxicity to aquatic invertebrates

Following experimental studies for the test chemical was summarized to conclude the toxicity of test chemical towards aquatic invertebrate is as follows:

The first experimental study was reviewed from Sewage works journal (1946) in this study the Short term toxicity test was carried out according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) with Daphnia magna as test organisms. The effective concentration (EC10) value of test chemical in Daphnia magna in a 24 hr study on mortality effect was determined to be 304 mg/L. Thus, considering the CLP Criteria for aquatic classification of the substance, it is concluded that test substance does not exhibit short term toxicity to aquatic invertebrate (Daphnia magna).

The next study Daphnia sp., Acute Immobilization experiment was performed according to OECD Guideline 202, for this the stock solution was prepared by dissolving 100 mg of the test substance in 100 ml of ADaM’s media (prepared in MilliQ water). A 25 mL of glass beaker was used to perform the experiment which was filled with 20mL of media including the test chemical. The test chemical was exposed to daphnia at a limit concentration of 100 mg/L for a period of 48 hours in a static condition. The median effect Concentration (EC50) of test chemical on Daphnia magna was determined as >100 mg/L on the basis of mobility of the Daphnia magna. Based on the EC50 value it can be consider that the test chemical is nontoxic and can be classified under the category of “not classified chemical” as per the CLP classification criteria.

To support the above studies another study was reviewed from Ecotoxicology and Environmental Safety journal in this study the short term toxicity test was carried out for 48 hr using daphnia magna as a test organism. Lethal Threshold Concentration value of test chemical in Daphnia magna in a 48 hr study on intoxication effect was determined to be 1450 mg/L. Thus, considering the CLP Criteria for aquatic classification of the substance, it is concluded that test substance does not exhibit short term toxicity to aquatic invertebrate (Daphnia magna)

Thus based on the overall studies the test chemical was concluded to be nontoxic and not classified as per the CLP classification criteria.

 

Long-term toxicity to aquatic invertebrates

Based on the prediction done using the EPI Suite ECOSAR version 1.11, the long term toxicity on daphnia was predicted for test chemical. On the basis of effects observed in a static freshwater system, the effect concentration NOEC value for the substance is estimated to be 4.126 mg/l for daphnia species for 21 days duration. Based on this value, and the test chemical is readily biodegradable it can be concluded that the test substance is considered as non-toxic  to daphnia magna at environmentally relevant concentrations and can be considered as "not classified"  as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria

Various experimental of test chemical and its read across chemicals have been reviewed to determine the toxicity of target to aquatic algae and their results are summarized below.

 

The first Experimental study performed for the target chemical and it was designed to assess the effect of the test substance on the growth of green algaChlorella vulgaris. The study was conducted in accordance with “OECD guideline for testing of chemicals No. 201 – Alga growth inhibition test”.

The effect of test substance was studied on the growth of fresh water green algaChlorella vulgaris.The study was conducted following OECD guideline 201- Alga growth inhibition test. The test concentration chosen for the study were 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l and 200 mg/l were prepared using stock solution of the test substance using de-ionized water. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of test substance. EC50 calculated from probit analysis was observed to be 75.25 mg/L. As the test chemical is readily biodegradable, it cannot be classfied as per CLP classification category.

 

Next experimental study was reviewed from handbook in this the green algae were exposed to the test concentration for a period of 14 days to observe average specific growth rate. On the basis of growth rate EC50 value was determined to be 60 mg/L for 3 hrs and > 10 mg/L for 14 days.

 

 Last experimental study was also reviewed from handbook and this study was designed to assess the effect of the test substance on the growth of green algae Haematococcus pluvialis. The green algae were exposed to the test concentration for a period of 4 hr to observe average specific growth rate. On the basis of growth rate EC10 value was determined to be165 mg/L .

 

By considering results of all the studies mentioned above it is concluded that EC50 value of test chemical is in range from 60 mg/L to 165 mg/L. On the basis of this range value and ready biodegradability test chemical is considered to be non toxic and cannot be classified as per CLP regulation.

Toxicity to microorganisms

In different studies, the given test chemical and structually and functionally similar read across chemicals has been investigated for toxicity to microorganisms to a greater or lesser extent. The studies are summarized as below –

 

In the first key study evaluation of toxicity of test chemical on Staphylococcus aureus toxicity to micro-organisms test was carried for 24 h under static condition.An overnight culture of each organism S. aureus was prepared. The 0.1 ml of organism was taken into 9.9 ml of sterile distilled water (SDW) to give 10 ml of 1:100 (10 ) dilution. The stock was maintained on nutrient agar slant and sub-cultured in nutrient broth for incubation at 37 °C prior to each antimicrobial testing. Inoculation of the test organisms on nutrient agar-prepared plates was achieved by flaming a wire loop on a spirit lamp, cooling the wire loop (air cooling) and fetching the test organisms.The discs were prepared using a Grade No. 1 Whatman filter paper. One hundred discs were obtained by punching and putting in vial bottles and sterilizing in an oven at 150 °C for 15 min. Thereafter the cups (9 mm diameter) were aseptically bored into the solid nutrient agar using a sterile cork-borer. The test solution of Methyl salicylate was introduced .The plates were left at room temperature for 2 h, allowed to diffuse into the medium, turned upside-down and thereafter incubated at 37 °C for 24 h in an incubator. The minimum inhibition concentration (MIC) of test chemical on Staphylococcus aureus was observed to be 200 mg/l.

First study was supported by the second study from peer reviewed journal. To evaluate antibacterial activity of  test chemical on Activated sludge toxicity to micro-organisms test was carried out according to the OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test. The test was carried for 3 h under static condition. Test chemicalwas used as a test material to evaluate toxicity to microorganisms as per OECD guideline 209. Activated sludge was used as the test culture at pH 7.5 for 3 h The EC50 value on the basis of respiration inhibition test was observed to be >1000 mg/l.

Thus based on the above effects chemical toxicity value ranges from 200 to > 1000 mg/l.