2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4)

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Endpoint:

toxicity to microorganisms, other

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data of read across chemical

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

WoE report is prepared based on toxicity to microorganisms study:1st, 2nd study

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4)- Molecular formula: C26H25N5O19S6.4Na- Molecular weight: 995.85 g/mole- Smiles : [Na][Na][Na][Na]Nc1ccc2c(cc(S(O)(=O)=O)c(N=Nc3ccc(S(=O)(=O)CCOS(O)(=O)=O)cc3)c2O)c1N=Nc1ccc(S(=O)(=O)CCOS(O)(=O)=O) cc1S(O)(=O)=O- Substance type: Organic- Physical state: Solid powder

Analytical monitoring:

not specified

Details on sampling:

1. Details on samplingConcentrations: final concentration 1,000 ppmSampling method: No data availableSample storage conditions before analysis: No data availableStudy 2:Concentrations: The test chemical conc. used for the study was 1000 mg/l (0.1%) and 10000 mg/l (1%)

Vehicle:

no

Test organisms (species):

other: 1. E. coli, Bacillus substilis, Aspergillus ochraceus and Penicillium ochrochloron MTCC 517. 2nd Paramaecium caudatum

Details on inoculum:

1. Laboratory culture: Aspergillus ochraceus NCIM 1146 was obtained from National Chemical Laboratory, Pune, India. E. coli MTCC 452, B. subtilis MTCC 6910 and Penicillium ochrochloron MTCC 517 were obtained from Microbial Type Culture Collection and Gene Bank (MTCC), Institute of Microbial Technology, Chandigarh, India.Method of cultivation: It was regularly maintained and preserved at 4°C on nutrient agar slants contained in (g/l); bacteriological peptone 10.0, beef extract 10.0 and NaCl 5.02nd study:Method of cultivation: Food dye (test chemical) of 0.1% conc. was put in a hollow slide glass, and an equal volume of 0.04 M phosphate buffer, pH 7.0, was added. After 5 to 10 Paramecium caudatum were added, their survival times were measured microscopically. 30 to 40 test organism for each conc. were tested and the mean survival time and the death rate was calculated.

Test type:

static

Water media type:

not specified

Total exposure duration:

20 min

Remarks on exposure duration:

no data

Post exposure observation period:

After 20 mins, the mean survival time and the death rate was calculated.

Nominal and measured concentrations:

2. nominal concentrations

Details on test conditions:

1. TEST SYSTEMTest vessel: discs of diameter 10mm2. Test vessel: Hollow slide glassNo. of organisms per vessel: 30 to 40 test organism for each test conc. was taken for the study.EFFECT PARAMETERS MEASURED (with observation intervals if applicable): After 20 mins, the mean survival time and the death rate was calculated.TEST CONCENTRATIONSTest concentrations: 10000 mg/l (1%) and 1000 mg/l (0.1%)

Reference substance (positive control):

not specified

Key result

Dose descriptor:

IC0

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: Aspergillus ochraceus, Penicillium ochrochloron MTCC 517

Dose descriptor:

other: MIC

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: E. coli

Remarks:

1st study

Study 1:

Table 1: Microbial toxicity of test chemical and its metabolites obtained after degradation

 

Micro-organisms	Diameter of the Zone of Inhibition (mm)
	Test chemical	Metabolites
E. coli	15.0	NI
Bacillus substilis	14.0	NI
Aspergillus ochraceus	NI	NI
Penicillium ochrochloron MTCC 517	NI	NI

NI: No inhibition

The diameter of the discs used was 10mm

2. The mean survival time (in sec) of test organism Paramecium caudatum was determined to be 695 seconds.

 

Table 1:EFFECT OF FOOD DYES ON THE MEAN SURVIVAL TIME AND DEATH RATE OF Paramecium caudatum

 

Test chemical	Dye concentration
	1.0%		0.1%
	Mean survival time (sec)	Death rate (%)	Mean survival time (sec)	Death rate (%)
Test chemical	695	77.4	-*	3.3

-* indicates no deaths were observed for 20 minutes

Validity criteria fulfilled:

not specified

Conclusions:

1. This investigation was aimed at identifying the effects of the test chemical and its degradation products on microbial growth. Test chemical and its degradation products were not toxic to Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 at 1,000 ppm concentration. 2. The death rate of the test organism at 10000mg/l was 77.4%. Therefore the Effective concentration causing more than 50% death of Paramecium caudatum is reported as 10000 mg/l

Executive summary:

Various studies available for the test chemical were reviewed to determine the toxic nature of 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS no 607724 -37 -8) on the growth of microorganisms. The studies are as mentioned below:

In the first study, this investigation was aimed at identifying the effects of the test chemical and its degradation products on microbial growth. Test chemical was purchased from Hi-media Laboratories Pvt. Ltd., Mumbai, India. Aspergillus ochraceus NCIM 1146 was obtained from National Chemical Laboratory, Pune, India. E. coli MTCC 452, B. subtilis MTCC 6910 and Penicillium ochrochloron MTCC 517 were obtained from Microbial Type Culture Collection and Gene Bank (MTCC), Institute of Microbial Technology, Chandigarh, India. It was regularly maintained and preserved at 4°C on nutrient agar slants contained in (g/l); bacteriological peptone 10.0, beef extract 10.0 and NaCl 5.0 Microbial toxicity of control and metabolites obtained after its decolorization (final concentration 1,000 ppm) was carried out in relation to E. coli, Bacillus substilis, Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 and zone of inhibition (diameter in mm) was recorded. The diameter of the discs used was 10mm. Test chemical and its degradation products were not toxic to Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 at 1,000 ppm concentration.

Similarly in the second study the death of Paramecium caudatum (PC), a unicellular animal, can be observed more readily and in far less time than that of small animals. Hence a bioassay was conducted to study the toxic effect of test chemical. Paramecium Caudatum was maintained at 22°C on 0.15 % dried lettuce infusion and fed with Aerobacter aerogenes. Chemical was tested in 0.1% and 1% concentration. The test concentrations were put in a hollow slide glass, and an equal volume of 0.04 M phosphate buffer, pH 7.0, was added. After 5 to 10 test organisms were added, their survival times were measured microscopically. Thirty to forty test organisms for each concentration were tested by the same method, and the mean survival time and the death rate were calculated. The survival time was defined as the time required until death was observed for each concentration. Death was assumed to have occurred when there was no movement. The death rate was defined as the percentage of deaths observed during 20 minutes. The mean survival time (in sec) of test organismParamecium caudatumwas determined to be 695 seconds.  The death rate of the test organism at 10000mg/l was 77.4%. Therefore the Effective concentration causing more than 50% death of Paramecium caudatum was reported as 10000 mg/l.

Thus based on the above studies, 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]diazenyl]-, sodium salt (1:4) (CAS no 607724 -37 -8) consider to be nontoxic and not classified as per the CLP classification criteria.

Description of key information

1. This investigation was aimed at identifying the effects of the test chemical and its degradation products on microbial growth. Test chemical and its degradation products were not toxic to Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 at 1,000 ppm concentration.

2. The death rate of the test organism at 10000mg/l was 77.4%. Therefore the Effective concentration causing more than 50% death of Paramecium caudatum is reported as 10000 mg/l

Key value for chemical safety assessment

EC10 or NOEC for microorganisms:

1 000 mg/L

Additional information

Various studies available for the test chemical were reviewed to determine the toxic nature of 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS no 607724 -37 -8) on the growth of microorganisms. The studies are as mentioned below:

In the first study, this investigation was aimed at identifying the effects of the test chemical and its degradation products on microbial growth. Test chemical was purchased from Hi-media Laboratories Pvt. Ltd., Mumbai, India. Aspergillus ochraceus NCIM 1146 was obtained from National Chemical Laboratory, Pune, India. E. coli MTCC 452, B. subtilis MTCC 6910 and Penicillium ochrochloron MTCC 517 were obtained from Microbial Type Culture Collection and Gene Bank (MTCC), Institute of Microbial Technology, Chandigarh, India. It was regularly maintained and preserved at 4°C on nutrient agar slants contained in (g/l); bacteriological peptone 10.0, beef extract 10.0 and NaCl 5.0 Microbial toxicity of control and metabolites obtained after its decolorization (final concentration 1,000 ppm) was carried out in relation to E. coli, Bacillus substilis, Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 and zone of inhibition (diameter in mm) was recorded. The diameter of the discs used was 10mm. Test chemical and its degradation products were not toxic to Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 at 1,000 ppm concentration.

Similarly in the second study the death of Paramecium caudatum (PC), a unicellular animal, can be observed more readily and in far less time than that of small animals. Hence a bioassay was conducted to study the toxic effect of test chemical. Paramecium Caudatum was maintained at 22°C on 0.15 % dried lettuce infusion and fed with Aerobacter aerogenes. Chemical was tested in 0.1% and 1% concentration. The test concentrations were put in a hollow slide glass, and an equal volume of 0.04 M phosphate buffer, pH 7.0, was added. After 5 to 10 test organisms were added, their survival times were measured microscopically. Thirty to forty test organisms for each concentration were tested by the same method, and the mean survival time and the death rate were calculated. The survival time was defined as the time required until death was observed for each concentration. Death was assumed to have occurred when there was no movement. The death rate was defined as the percentage of deaths observed during 20 minutes. The mean survival time (in sec) of test organismParamecium caudatumwas determined to be 695 seconds.  The death rate of the test organism at 10000mg/l was 77.4%. Therefore the Effective concentration causing more than 50% death of Paramecium caudatum was reported as 10000 mg/l.

Thus based on the above studies, 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]diazenyl]-, sodium salt (1:4) (CAS no 607724 -37 -8) consider to be nontoxic and not classified as per the CLP classification criteria.