4-amino-5-hydroxynaphthalene-1,7-disulphonic acid

Endpoint summary

Additional information

Short term fish toxicity:

By applying the weight of evidence approch to Short term fish toxicity study for Target -7-Amino-1-hydroxy-3-naphthalenesulfonic acid(Cas no.130-23-4)Summarized as fallowed:

Study from SSS QSAR prediction model, 2016 for target chemical 4-amino-5-hydroxynaphthalene-1,7-disulphonic acid estimates the short term toxicity to fish -Oryzias latipes at 96 hours LC50 which is to be 294.099151611 mg/l in semi static condition and with mortality effects.

Above target prediction supported by read across chemical (Cas no.87-02) from peer reviewed study(GREIM H. et al;Chemosphere; Vol. 28, No. 12, pp. 2203.2236, 1994)The investigation was performed to determine the toxicity of sulphonic acids to fish the lethal concentration for test substance 7-Amino-4 hydroxy- 2 – naphthalenesulphonic acid in short term fish toxicity (LC0) was determined to be 1000 mg/l. At this dose

concentration mortality was not observed.

This LC0 value indicate that the test substance 7-Amino-4 hydroxy- 2 – naphthalenesulphonic acid is non toxic to fish upto dose concentration 1000 mg/l.

 

Another study of read across substance (Cas no. 90-51-7) fromStudy report of Vernon C. Applegate,;Special Scientific Report- Fisheries No. 207;1957 as theshort term toxicity test to rainbow trout, bluegill and sea lamprey was conducted for 24 hrs.

Test organisms Sea lamprey (Petromyzon marinus),Rainbow trout (Salmo gairdnerii) and Bluegill (Lepomis macrochius) were used for the study.The study was performed under static conditions for 24 hrs. The test chemical conc. used for the study was 5 mg/l.

 Larval lampreys were collected by means of an electric shocker in the Ocqueoc River, Presque Isle County, Michigan, and were held in running water in aquaria and small “races” under conditions which simulated their natural stream habitat.

  Test fishes (Rainbow trout and Bluegill) were obtained fromthe stocks of local state and Federal fish hatcheriesand were held in large raceways. These specimens were maintained in the best possible physical condition until used in the laboratory.

 

The aggregate test animals available, usually six in number, were placed together in a 10-literglass battery jar containing 5 liters of water. The jars were provided with aeration through standard stone air-breakersand were maintained at a constant temperature by immersion in specially constructed constant temperature troughs. Water temperature was maintained within the limits of ± 1.0 °F.

 

Twelve of these test jars (each containing a substance being assayed) were included with one control jar in each trough. Fish and larvae in the control jar were exposed only to the water and physical conditions of the typical test container.

Test condition contains dissolved oxygen from 8.6 to 13.7 ppm and free CO₂from 5.0 to 9.0 ppm

Observations of each test were measured approx. six times, at various intervals, during the 24 hr test period. At each observation, the condition of every test specimen was determined and recorded. Chronological histories were thus obtained of any symptoms of illness and the occurrence of death.

 No effects were observed when the test organisms sea lamprey, bluegill and rainbow trout were exposed to the test chemical for 24 hrs. Thus, the NOEC value was found to be 5 mg/l.

 

In other experimental data for same read across Data bank of Environmental Properties of Chemicals (EnviChem);2014 performshort term toxicity to fish the 48 hrs LC50 value to test fishes was found to be 335 mg/l.This LC50 value indicate that the test substance 2-Amino-8-naphthol-6-sulfonic acid is non toxic to fish as per the CLP criteria.(i.e value >100 mg/l)

 

 

Thus,Overall weight of evidence studies from prediction model for target chemical (Cas no.130-23-4) and peer reviewed journals of read across substances (Cas no.87-02-5 and 90-51-7) indicate that the chemical 7-Amino-1-hydroxy-3-naphthalenesulfonic acid is non toxic to fish as per CLP criteria (i.e LC50 >100mg/l)and thus not consider for further classification.

Toxicity to aquatic algae and cyanobacteria

Various studies for the target chemical4-Amino-5-hydroxynaphthalene-1,7-disulphonic acid including the predicted data and its read across substance were reviewed to summarize the following information:

 

The effect of test item disodium 4-amino-5-hydroxynaphthalene-1,7-disulphonic acid, CAS No. 130-23-4 was studied [UERL Study Report, Sustainability Support Services (Europe) AB (Report no. 130-23-4/01/2016/AT), 2016]on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 6.25mg/l, 12.5mg/l, 25mg/l, 50mg/l,100mg/l and 200mg/l. The test concentrations were prepared using stock solution of the test item using mineral media. 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 the test item. EC50 calculated graphically through probit analysis was observed to be >200mg/l.

 

 

72 hrs aquatic toxicity study (Danish (Q)SAR Database, 2016) was conducted to assess toxic effects of the test compound4-Amino-5-hydroxynaphthalene-1,7-disulphonic acid(CAS no 130-23-4) and the results were predicted. The study was based on the effects of the test compound on Pseudokirchneriella spp. in a static fresh water system. The predicted data suggests the effective concentration (EC50) for the test compound4-Amino-5-hydroxynaphthalene-1,7-disulphonic acid(CAS no 130-23-4) was estimated to be 39583.34 mg/L on the basis of growth rate.

 

 

96 hrs aquatic toxicity study (EPI suite, 2016) was conducted to assess toxic effects of the test compound4-Amino-5-hydroxynaphthalene-1,7-disulphonic acid(CAS no 130-23-4) and the results were predicted. The study was based on the effects of the test compound on green algae in a static fresh water system. The predicted data suggests the effective concentration (EC50) for the test compound4-Amino-5-hydroxynaphthalene-1,7-disulphonic acid(CAS no 130-23-4) was estimated to be 111000 mg/L on the basis of growth rate.

 

 

Short term toxicityto Chlorella pyrenoidosa(green algae) study was carried out for 72 hrs (JU-CHANG HUANG and EARNEST F. GLOYNA, 1968)

Emerson strain of bacteria free, experimentally reproducible cultures ofChlorella pyrenoidosawas used as a test organism. The procedure involve the use of test tubes in both the screening and final tests. These test tubes contained 15 ml of inorganic culture medium, a predetermined amount of test chemical and 5 ml of algal culture. The tubes were incubated for 72 hrs and chlorophyll content of the algal suspensions was measured every 24 hrs.For chlorophyll measurement, the chlorophyll pigment was extracted with hot methanol in two separate extractions. An algal suspension, 2.5 ml, was removed from the test tube, centrifuged, washed with distilled water, and recentrifuged in preparation of chlorophyll analysis. After discarding the supernatant, the deposited cell material was coagulated by placing the cells in a boiling water bath for about 40 sec. About 2.5 ml of methanol were used in each extraction. Finally, the chlorophyll solution was diluted to a total volume of 10 ml with an acetone-water mixture (80 per cent by volume).A Beckman Spectrophotometer, Model DB, was used to measure the chlorophyll content. For this a wavelength of 652 m/z was used because different proportions of chlorophyll a and b least affect the results at this wavelength.Control tubes containing no test chemical was also used in the experiment.Knop's solution, including the Hutner-EDTA microelement addition, was used as the culture medium. pH of culture medium was adjusted to 7.0 using KOH before use. The test organism was maintained under steady-state conditions, provided a chlorophyll content of 38 mg/l. Environmental control was rigidly maintained. The temp. of water bath was 25 ± 1°C.The test apparatus consisted of a constant-temperature water bath, a light source containing four 200W fluorescent lamps with attached aluminum reflectors, a gas manifold to supply an air-CO2 mixture to each test tube, and a rack to hold the test tubes. A stream of 5 % CO2 in air gas mixture was supplied to culture medium in order to provide the inorganic carbon source and also to keep the algal ceils in suspension.

 

Based on destruction of chlorophyll of test organism by test chemicalAmino-1-phenol-4-sulfonic acid, the LOEC value was found to be1500 mg/l and as no toxic effect at 1000 mg/l was observed, the NOEC value was found to be 1000 mg/l.

 

 

 

Short term toxicityto Chlorella pyrenoidosa(green algae) study was carried out for 72 hrs (JU-CHANG HUANG and EARNEST F. GLOYNA, 1967).

 

Emerson strain of bacteria free, experimentally reproducible cultures ofChlorella pyrenoidosawas used as a test organism. An Emerson strain of bacteria-free, experimentally reproducible cultures ofChlorella pyrenoidosawas obtained from Dr.Jack E. Myers, Professor of Zoology and Botany and Director, Laboratory of Algal Physiology, The University of Texas.Five conc. levels of test chemical were used for the tests (100, 500, 1000, 1500 and 2000 mg/l, respectively).A test tubes was used in the screening and final tests. These test tubes contained 15 ml of inorganic culture medium, a predetermined amount of test chemical and 5 ml of algal culture. The tubes were incubated for 72 hrs and chlorophyll content of the algal suspensions was measured every 24 hrs.Control tubes containing no test chemical in the culture medium was also used in the experiment.Environmental control was rigidly maintained. The temp. of water bath was 25 ± 1°C.The special toxicity test device consisted of an aquarium complete with a light source, a gas manifold and an apparatus for holding the test tubes. In this test device, algal cultures were kept under a rigid environmental control. A stream of 5 % CO2 in air gas mixture was supplied to culture medium in order to provide the inorganic carbon source and also to keep the algal ceils in suspension.Screening tests were performed to establish the threshold toxic concentration as well as completely algicidic one. Knop's solution, including the Hutner-EDTA microelement addition, was used as the culture medium. pH of culture medium was adjusted to 7.0 using KOH before use.Kimble Product of plain culture tubes, Series 45060, was used as the test tube. These test tubes have the size of 19 x 150 mm with a volume of 25 mI.The initial algal cell density was1.0 gm/l dry weight or equivalent to 3.8 cm/ml packed cell volume and chlorophyll content was 38 mg/l.The chlorophyll pigment was extracted with hot methanol from a 2.5ml algal suspension which was pipetted out of the test tube. The algal cells were centrifuged out of the culture medium, washed with distilled water and then centrifuged again. The supernatant was discarded and the remaining packed cell material was coagulated in a boiling water bath for a period of about 30 to 45 seconds depending on the amount of cells. Thereafter, the chlorophyll was extracted twice by using hot methanol.The extracted chlorophyll solution was then dissolved in 80 percent by volume acetone-water mixture to make a final total volume of 10 ml. The chlorophyll content was then analyzed under the spectrophotometer. Content of all extracted chlorophylls which were dissolved in 10mlof 80 percent acetone-water mixture could be determined by measuring the optical density (O.D.) at a wavelength of 652 mn. This wavelength was optimum because various proportions of chlorophylls a and b would least affect the result.A Beckman Spectrophotometer, Model DB, was used for all chlorophyll analysis. This spectrophotometer is a double beam instrument for making transmittance and absorbance measurements in the 205 to 770mn wavelength range.

 

Based on the decrease in chlorophyll content of test organism by test chemicalAmino-1-phenol-4-sulfonic acid, the LOEC value was found to be2000 mg/l and as no toxic effect at 1000 mg/l was observed, the NOEC value was found to be 1000 mg/l.

 

 

On the basis of the above results of various studies for target chemical and its read across substance, it can be concluded that the substance can be considered as non- hazardous to aquatic organisms.

 

Toxicity to microorganisms study for Target -7-Amino-1-hydroxy-3-naphthalenesulfonic acid(Cas no.130-23-4)Summarized as fallowed:

 

From SSS QSAR prediction model, 2016 for target chemical:

The toxicity to microorganism was estimated by the QSAR Toolbox version 3.3 prediction,in freshwater and semi static condition the 48 hours IGC50(Inhibition growth concentration) was estimated to be 823.090393066 mg/l for test substance 4-amino-5-hydroxynaphthalene-1,7-disulphonic acid with growth inhibition effects.

This predicted IGC50 value indicate that the test substance 4-amino-5-hydroxynaphthalene-1,7-disulphonic acid is non hazardous to aquatic microorganisms.

Other peer reviewed summary (GREIM H.,et al;Chemosphere. Vol. 28, No. 12, pp. 2203.2236, 1994) of read across chemical 7-Amino-4 hydroxy- 2 – naphthalenesulphonic (Cas no.87-02-5) as followed:

The investigation was performed to determine the toxicity of sulphonic acids to bacteria.

Based on different test procedures, The EC0 for 7-Amino-4 hydroxy- 2 – naphthalenesulphonic Acid was determined to be 1000 mg/l on the basis of inhibition effect.

Based on the no effect concentration value the test chemical 7-Amino-4 hydroxy- 2 - naphthalenesulphonic Acid can be considered non- toxic to bacteria (i.e microorganisms).

 

Another result from experimental study- Journal of Scientific & Industrial Research, Vol. 70, Pg. no. 525-532, July 2011. Of read across substance (Cas no.90-51-7):

Toxicity to micro-organisms study was conducted on gram positive bacteria (Staphylococcus aureus,Bacillus subtilis), gram negative bacteria (Escherichia coliandPseudomonas aeruginosa) and fungiCandida albicans.

 

The antibacterial assay was performed using Kirby – Bauer disk diffusion method. MIC was determined for all the different test organisms.

 

Test chemical was dissolved in DMF (< 500 ppm conc.). The test substance conc. used for the study was 20, 50, 100, 200, 300, 500, and 1000 mg/l, respectively.

 

The melting point of test chemical was determined in PMP-DM scientific melting point apparatus. Purity of test chemical was determined by thin layer chromatography (TLC) using silica gel-G coated Al-plates (0.5 mm thickness).

 

Infrared spectra was recorded on Shimadzu FT - IR 8400S model using KBr pellets.¹H NMR spectra was acquired on a Varian 400 MHz model spectrophotometer using DMSO as a solvent and TMS as internal reference (chemical shifts in ‘, ppm).

 

Elemental analysis of C, H, and N were carried out on Carlo Erba 1108 instrument.

 

For antibacterial and antifungal assay, ciprofloxacin and flucanazole was used as a standard dyes.

The melting point of test chemical was found to be > 300°C.

  The MIC value for 5 different organism’s are-

Pseudomonas aeruginosa– 1000 mg/l,Staphylococcus aureus– 300 mg/l ,Escherichia coli– 300 mg/l ,Bacillus subtilis– 100 mg/land Candida albicans– 300 mg/l

All above MIC values for different species i.e gram positive bacteria (Staphylococcus aureus,Bacillus subtilis), gram negative bacteria (Escherichia coliandPseudomonas aeruginosa) and fungiCandida albicansindicate that the test substance is non toxic to microorganisms.

Overall weight of evidence studies from prediction model for target chemical (Cas no.130-23-4) and peer reviewed journals of read across substances (Cas no.87-02-5 and 90-51-7) indicate that the chemical 7-Amino-1-hydroxy-3-naphthalenesulfonic acid is non toxic to micro-organism and thus not consider for further classification.

Short term toxicity to aquatic invertebrates:

Various predicted studies for the target chemical 4-amino-5-hydroxynaphthalene-1, 7-disulphonic acid were reviewed to summarize the following information: 

48 hrs aquatic toxicity study (EPI suite, 2016) was conducted to assess toxic effects of the test compound 4-amino-5-hydroxynaphthalene-1, 7-disulphonic acid (CAS no 130-23-4) and the results were predicted. The study was based on the effects of the test compound on Daphnia magna in a static fresh water system. The predicted data suggests the effective concentration (EC50) for the test compound 4-amino-5-hydroxynaphthalene-1, 7-disulphonic acid was estimated to be778000 mg/lon the basis of mobility.

 

48 hrs aquatic toxicity study (SSS QSAR prediction model, 2016) was conducted to assess toxic effects of the test compound 4-amino-5-hydroxynaphthalene-1, 7-disulphonic acid (CAS no 130-23-4) and the results were predicted. The study was based on the effects of the test compound on the Daphnia magna in a static fresh water system. The predicted data suggests the effective concentration (EC50) for the test compound 4-amino-5-hydroxynaphthalene-1, 7-disulphonic acid (CAS no 130-23-4) was estimated to be 133.610977173 mg/L on the basis of mobility. 

  

On the basis of results of various predictions for toxicity to aquatic algae and cyanobacteria, it can be considered that the substance is not likely to be toxic to aquatic invertebrates at environmentally relevant concentrations and can be classified as non- hazardous as per the criteria of CLP regulation.