Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 12 September 2006 to 7 November 2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Objective of study:

bioaccessibility (or bioavailability)

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD guideline 474 (in vivo micronucleus)

Deviations:

no

Principles of method if other than guideline:

Rats were dosed once by oral gavage with 2000 mg/kg of test item. Blood was sampled at 15 minutes, 30 minutes and 1 hours (orbital sinus smear). The serum was isolated by centrifugation. It was theneafter centrifugated ain order to eliminate fibrine. A High Performance Liquid Chromatography : Ultra Violet (HPLC/UV) approach was used to quantified the amount of test item in serum (using a calbiration curve)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

EC Number:

224-618-7

EC Name:

Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

Cas Number:

4430-18-6

Molecular formula:

C21H15NO6S.Na

IUPAC Name:

sodium 2-[(4-hydroxy-9,10-dioxo-9,10-dihydroanthracen-1-yl)amino]-5-methylbenzenesulfonate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: supplied by l'Oreal, batch No. 10130
- Expiration date of the lot/batch: July 2007
- Purity test date: 28 July 2006

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:stored in glass flask at +4 deg Celsius, away from light, under inert gas
- Stability under test conditions: no information
- Solubility and stability of the test substance in the solvent/vehicle: solubility <1g/100 mL

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
All the concentrations and dose-levels were expressed as active item taking into account the purity of 59.8%. Before preparation, the vehicle was degassed by sonication for at least 15 minutes and then saturated with nitrogen gas, and kept under nitrogen atmosphere for 15 minutes. The test item dosage form was prepared extemporaneously under nitrogen atmosphere (under magnetic stirring) and protected from light. They were kept under nitrogen gas and protected from light before being used within 30 minutes/1 hour after preparation. The test item was ground to a fine powder using a mortar and pestle, dissolved in the vehicle in order to achieve the concentration of 100 mg/mL (expressed as active item) and then homogenized using a magnetic stirrer. Using a treatment volume of 20 mL/kg, the target dose-level was 2000 mg/kg.

Radiolabelling:

no

Test animals

Species:

rat

Strain:

Swiss

Remarks:

Swiss Ico : OF1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories (France)
- Age at study initiation: 6 weeks old
- Weight at study initiation: 30±2g for males ; 24±1g for females
- Assigned to test groups randomly: yes
- Fasting period before study: not specified
- Housing: housed by groups in polycarbonate cages. Each cage contained autoclaved
sawdust (SICSA,France)
- Diet (e.g. ad libitum): free access to A04 C pelleted maintenance diet (SSNIF, Germany)
- Water (e.g. ad libitum):Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum
- Acclimation period: at least 5 days before the day of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±1 deg Celsius
- Humidity (%): 30 to 70%
- Air changes (per hr): at least 12 cycles/hour of filtered non recycled fresh air
- Photoperiod (hrs dark / hrs light): 12h/12h (07h-19h)

IN-LIFE DATES: From: 5 September 2006 To: 21 September 2006

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

All the concentrations and dose-levels were expressed as active item taking into account the purity of 59.8%. Before preparation, the vehicle was degassed by sonication for at least 15 minutes and then saturated with nitrogen gas, and kept under nitrogen atmosphere for 15 minutes. The test item dosage form was prepared extemporaneously under nitrogen atmosphere (under magnetic stirring) and protected from light. They were kept under nitrogen gas and protected from light before being used within 30 minutes/1 hour after preparation. The test item was ground to a fine powder using a mortar and pestle, dissolved in the vehicle in order to achieve the concentration of 100 mg/mL (expressed as active item) and then homogenized using a magnetic stirrer. Using a treatment volume of 20 mL/kg, the target dose-level was 2000 mg/kg.

Duration and frequency of treatment / exposure:

Single treatment, 1 hours

Doses / concentrations

Dose / conc.:

2 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose / concentration:

3 animals per sex per sampling time group

Control animals:

no

Details on study design:

CRITERIA FOR DOSE SELECTION:
On the basis of a previous bone marrow mouse oral micronucleus test (CIT/Study No. 11042 MAS), which was conducted with the same compound up to the maximum dose of 2000 mg/kg (i.e. 1086 mg/kg when expressed as active item) and which was negative, a limit test at 2000 mg/kg (expressed as active item) was performed .

PRINCIPLE :
The serum samples were deproteinized by addition of methanol. The clear supernatant (containing the test item) was analyzed by reversed phase chromatography (HPLC) with detection at 560 nm.

Details on dosing and sampling:

TOXICOKINETIC / PHARMACOKINETIC STUDY (Absorption, distribution, excretion)
- Tissues and body fluids sampled (delete / add / specify): serum
- Time and frequency of sampling: 15 minutes, 30 minutes and 1 hours after treatment
- Method type(s) for identification HPLC-UV
- Limits of detection and quantification: 1µg/mL

Results and discussion

Main ADME results

Type:

other: Systemic exposure

Results:

These results clearly demonstrated the systemic exposure of treated animals to the test item.

Any other information on results incl. tables

Table 1 : Summary of the results	15 min µg/mL	30 min µg/mL	1 hour µg/mL
Males	8.61 ± 3.04	9.16 ± 0.63	7.89 ± 2.55
Females	25.7 ± 7.7	19.9 ± 4.1	20.4 ± 8.3

 

Applicant's summary and conclusion

Conclusions:

Under experimental conditions of this study, the Acid Violet 43 after oral gavage at 2000 mg/kg was found in serum of treated rats at each time point samples. The study showed a systemic exposure of rats after oral gavage.

Executive summary:

This GLP compliant study was performed to assess a potential systemic exposure in rats which were treated orally with the registered substance Acid Violet 43. This study was performed on satellite animals used for in vivo micronucleus assay.

3 rats per sex and per time point group were dosed orally by gavage. At 15 minutes, 30 minutes and 1 hour, blood was sampled in orbital sinus. Serum was isolated and deproteinsed. The amount of the test item was measured by a HPLC-UV method and calculated by interpolation with a calibration curve (made with known concentration curve) by linear regression analysis with the serum blood samples.

Under experimental conditions of this study, the Acid Violet 43 after oral gavage at 2000 mg/kg was found in serum of treated rats at each time point samples. The study showed a systemic exposure to the test item of rats after oral gavage. No clastogenic effects were reported.