Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics

Type of information:

other: Expert assessment

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

A written assessment of toxicokinetic behaviour is considered appropriate for the substance. The substance displays only minor toxicological effects in any of the studies proposed, and is deemed to be be not harmful for health effects. As such, it is deemed inappropriate in terms of animal welfare to conduct a toxicokinetic assessment when no harmful effects are predicted based on known toxicology. A written assessment has therefore been prepared to address this endpoint.

Materials and methods

Objective of study:

other: Assessment of toxicokinetic behaviour

Principles of method if other than guideline:

Written assessment based on toxicological profile.

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

EC Number:

413-550-5

EC Name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Cas Number:

142068-96-0

Molecular formula:

Hill formula: C53H28Cl2N14Na8O26S8

IUPAC Name:

octasodium 5-[(4-chloro-6-{[4-({[4-chloro-6-({7-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-8-hydroxy-3,6-disulfonatonaphthalen-1-yl}amino)-1,3,5-triazin-2-yl]amino}methyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-3-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate

Details on test material:

Not applicable

Test animals

Species:

other: Not applicable

Administration / exposure

Details on exposure:

Not applicable

Duration and frequency of treatment / exposure:

Not applicable

Doses / concentrations

Remarks:

Doses / Concentrations:
Not applicable

No. of animals per sex per dose / concentration:

Not applicable

Positive control reference chemical:

Not applicable

Details on study design:

Not applicable

Details on dosing and sampling:

Not applicable

Statistics:

Not applicable

Results and discussion

Metabolite characterisation studies

Metabolites identified:

not measured

Details on metabolites:

Not applicable

Any other information on results incl. tables

The following information about the toxicokinetics were based on examination of the toxicological test package and knowledge known historically about reactive dyes. Independent experimental studies on the toxicokinetics was not done.  Toxicokinetic parameters such as uptake, distribution, metabolism and excretion form the essential toxicological profile of a substance. The assessment of the toxicokinetic properties of Substance H 112339 given below is based on the results obtained for the following toxicological endpoints;

Acute oral toxicity

Acute dermal toxicity

Skin irritation

Skin sensitisation

Ames Test – salmonella& E.coli

In Vitro Cytogenetic Assay in Human Lymphocytes

In vitro gene mutation test in CHO cells (HPRT Locus Assay)

Subacute (28-day) oral toxicity

Sub-chronic (90-day) oral toxicity study

 

All studies were carried out according to the principles of Good Laboratory Practice and met the requirements of the OECD and EU-Guideline for the Testing of Chemicals. Simultaneously reference to physico-chemical data such as solubility in solvents, log Pow and hydrolytic stability is included

 

General

 

The degree of purity of the substance is 70-80 %.Unknown byproducts of the dyestuff synthesis include water, salts and sulphonic acid organic derivatives, which are given as impurities. 

 

Substance H 112339 is red powdered or granular solid at room temperature conditions. A vapour pressure determination performed and gave a value of <10-5 Pa at 25⁰C. Therefore a significant inhalation exposure to vapours is not expected. The low n-octanol/water partition coefficient (log Pow -5.7 at 20 ° C) as well as the relatively high molecular weight of the substance (>1600) a systemic bioavailability after dermal exposure is not anticipated.

 

Toxicological Profile:

 

Substance H 112339 was tested for acute oral toxicity in male and female Wistar rats. After application of 2000 mg/kg body weight by gavage, neither nor any clinical symptoms occurred. However, red staining of urine clearly indicates absorption and systemic availability of substance H 112339. Urinary excretion of Substance H 112339 being visualized by red staining of urine took about 4 days.

 

Based on the results of this study the median lethal dose (LD50) of Substance H 112339 in the rat is greater than 2 000 mg/kg body weight. Dermal treatment with 2 000 mg/kg body weight caused slight irritation and staining but caused no mortality or symptoms of toxicological relevance. Substance H 112339 is however irritating to the skin (but was reversible apart from staining) and is not a skin sensitiser in the Buehler test. According to the results of the acute dermal toxicity study as well as the skin irritation and sensitisation data and supported by the pronounced hydrophilic properties, Substance H 112339 most probably has no significant dermal absorptive potential.

 

Studies on genotoxicity (Ames-Test (ICJ CTL, 1991), in vitro cytogenetics (ICf CTL, 1992), in vitro HPRT-test (BASF AG, 2000)) were negative either with and without metabolic activation. Therefore, there is no indication of reactivity of Substance H112339 and its metabolites under the test conditions.

 

Most probably DNA reactive species are not formed in liver although liver enzymes lead to enhanced bacterial toxicity which suggests biotransformation activation of the substance.

 

In a 28-day study in rats with doses of 0, 50, 250 and 1000 mg/kg body weight given by gavage, clinical, clinico-chemical and histo-pathological effects observed in the 250 mg/kg and 1000 mg/kg groups also demonstrate absorption and systemic availability of Substance H 112339 (ICI Central Toxicology Laboratory, 1992) . In the 1000 mg/kg group (and in part also in the 250 mg/kg group), staining of tissues was observed at terminal and recovery kill. Since staining of tissues after two weeks of recovery is still present, excretion of Substance H 112339 being distributed in tissues appears to be rather slow. Staining of urine also indicates that renal excretion is a relevant pathway for the elimination of Substance H 112339.

 

In the 90-day study in rats, in which doses of 50. 250 and1000 mg/kg body weight were administered by gavage, clinical, clinico-chemical and (histo-)pathological effects were observed at all dose levels in females and at doses ≥ 250 mg/kg body weight with the kidneys being the main target organ (BASF AG, 2000).  The results of this 90-day study confirm the statements on the toxicokinetic behaviour of Substance H 112339 in that:

 

a) Substance H 112339 is absorbed from the gastrointestinal tract and is systemically available in all tissues and organs

b) Substance H 112339 and its metabolites is - at least in part - excreted by the kidneys

 

In a 1-generation reproduction study in the rat, no effects on reproduction were noted, indicating that prolonged exposure is not expected to cause reproductive effects. 

 

Evaluation :

 

Based on all available data, Substance H 112339 does not exhibit a conspicuous toxicokinetic behaviour. Substance H 112339 has a very low acute toxicity potential. The results from all studies with dermal exposure indicates that Substance H 112339 has insignificant or no dermal absorptive potential. Additionally, taking the results of the subacute and subchronic oral toxicity study into account, H112339 is absorbed from the gastrointestinal tract and is systemically available in all tissues and organs.  

 

As such, a significant bioaccumulation potential in mammals can most probably be excluded, particularly given the low solubility in fat (0.009mg/100g fat), despite the observations in the studies. This conclusion is further endorsed by the effects of a bioaccumulation study in fish which clearly indicated that the substance is not bioaccumulated in this species (BCF < 3.3) Toxicity was further characterized by hypoplastic anemia with corroborating histopathological findings in spleen and bone marrow. Furthermore, as a consequence of severe systemic toxicity, also liver, ovaries, adrenal cortex, and serous coats were affected. By contrast, the discoloration of urine, feces, and/or skin was related to the tinctorial properties of the test substance (dye-stuff), but does not represent a toxicologically relevant finding.

 

The slight changes in kidney pathology and/or urinalysis of individual low dose females can be interpreted as remnants of transient tubular degeneration. The slight decrease in hematocrit in females of this dose level is assessed as being a mild adverse effect, only due to the lack of a histopathological correlate or other concomitant changes in red blood cells.

 

 From the mutagenicity assays it appears that Substance H 112339 is not metabolised toward genotoxic structures.

   

Summary:

The results of basic toxicity testing give no reason to anticipate unusual characteristics with regard to the toxicokinetics of Substance H 112339 . The data indicate that there is little or no dermal absorption. Although substance specific toxicity can be attributed to the material in prolonged exposure studies, significant bioaccumulation of Substance H 112339 can most probably be excluded due to the marked hydrophilic properties and lack of solubility in fat. Based on the negative mutagenicity assays, a metabolisation towards genotoxic sub-structures can be ruled out.

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): other: no bioaccumulation potential based on review of study results

Executive summary:

The results of basic toxicity testing give no reason to anticipate unusual characteristics with regard to the toxicokinetics of Substance H 112339 . The data indicate that there is little or no dermal absorption. Although substance specific toxicity can be attributed to the material in prolonged exposure studies, significant bioaccumulation of Substance H 112339 can most probably be excluded due to the marked hydrophilic properties and lack of solubility in fat. Based on the negative mutagenicity assays, a metabolisation towards genotoxic sub-structures can be ruled out.