3,6-bis(diethylamino)-9-[2-(methoxycarbonyl)phenyl]xanthylium tetrachlorozincate

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary source.

Data source

Reference

Reference Type:

secondary source

Title:

Repeated dose oral toxicity study of the test chemical

Author:

Biodynamics Incorporation

Year:

1992

Bibliographic source:

NTRL

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

A Three-month Dose Range-Finding Study of test chemical in Mice was conducted to evalute its toxic nature.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthylium chloride

EC Number:

201-383-9

EC Name:

9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthylium chloride

Cas Number:

81-88-9

Molecular formula:

C28H31N2O3.Cl

IUPAC Name:

9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthenium chloride

Constituent 2

Reference substance name:

Rhodamine B

IUPAC Name:

Rhodamine B

Details on test material:

- Name of test material: D&C Red 19
- Molecular formula: C28H31N2O3Cl
- Molecular weight: 479.017 g/mol
- Substance type: Organic
- Physical state: No data
- Impurities: No data

Test animals

Species:

mouse

Strain:

other: Charles River CD-1, COBS (ICR derived)

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Charles River Greeding Laboratories Wilmington, Massachusetts
- Age at study initiation: Weanlings (4-5 weeks)
- Weight at study initiation:
Mean Range
Male; 25 (16-31)
Female:22 (10-29)

- Housing: Individually in elevated stainless steel wire mesh cages.
Food-Standard laboratory diet (Puri na
Laboratory ) ad libitum. Fresh food presented twice weekly.
Water-Ad libitum

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: Diet

Details on oral exposure:

Appropriate amounts of the test chemical were mixed fresh weekly with standard 1aboratory diet. Diet analysis: Four ounce samples of the control and each dose level were collected weeks 1. 4. 8 and 13. and analysis was conducted by the Environmental and Analytical Chemistry Division of Bio/dynamics. Inc. Storage temperature: Room temperature

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Approximately one ounce of pure (compound was collected weeks 1, 4, 8 and 13; analysis conducted by the Environmental and Analytical Chemistry Division of Bio/dynamics, Inc.

Duration of treatment / exposure:

90-91 days

Frequency of treatment:

(Daily) Continuously in the diet

Doses / concentrations

Remarks:

Doses / Concentrations:
0.001. 0.01, 0.1, 1.0 and 5.0%=1.66, 16.66, 166.66, 1666.66, 8333.33mg/kg bw.
Basis:

No. of animals per sex per dose:

120 mice (60 males; 60 females).

Control animals:

yes, concurrent vehicle

Details on study design:

No data

Positive control:

No data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
Twice pretest, weekly during treatment, and terminally (after fasting) including organ weights of brain kidneys liver, spleen

Sacrifice and pathology:

GROSS PATHOLOGY: Yes Twice daily
HISTOPATHOLOGY: Yes

Other examinations:

No data

Statistics:

Statistically significant differences from control are indicated in mean tables and appendices.

Results and discussion

Results of examinations

Clinical signs:

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

All males and females receiving 5.0: pure color died spontaneously during the first or second week of test substance administration. In addition, two males receiving 1.0: pure color died spontaneously during the course of the study. Five animals one from each of groups I through V, died accidentally on day 76 or 77 due to starvation.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Mean body weights for males and females receiving 1.0% pure color were generally lower than those of the control animals from week 1 through termination all differences were statistically significant and greater than 10%.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Elevations, as compared to control, were noted in the abso1ute and relative (to body weight) mean liver weights for both sexes receiving 0.1 and 1.0% pure color all weights were significantly (p≤0.01) different. As compared to control, absolute mean spleen and kidney weights were, reduced and relative mean brain weights were elevated for both sexes receiving 1.0% pure color, .however these differences were considered a reflection of the lower mean
body weights for this group.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

The mucosa of the gastrointestinal tract and the bila were tinted red in many animals at dose levels of 0.1 and 1.0%. None of the other changes observed in animals at the terminal sacrifice, nor in animals dying spontaneously or accidentally were considered related to the administration of D&C Red '19 at any dose level.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not specified

Description (incidence and severity):

Animals at the 0.1% level had a mild diffuse periportal hepatocellular hypertrophy with an increase in nuclear and cytoplasmic mass and narrowing of sinusoidal spaces considered to be test substance-related. Other changes were considered to be spontaneous in origin and not related to D&C Red #19 administration.

Other effects:

not specified

Effect levels

Dose descriptor:

NOAEL

Effect level:

166.6 mg/kg diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Hepato-toxicity and variation in body weights

Remarks on result:

other: No toxic effects were observed .

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The No observed adverse effect level (NOAEL) was considered to be 166.66 mg/kg for test chemical

Executive summary:

The study was conducted to determine the Maximum tolerated dose of test chemical in Charles river CD-1 mice .When administered Orally via  the diet at dose levels of 0.001, 0.01, 0.1, 1.0 and 5.0% (1.66, 16.66, 166.66, 1666.66, 8333.33mg/kg bw/day)pure color for a period of 90-91 days. Animals were observed for mortality,clinical sign, food consumoption,organ weight,gross and histipathology.All animals receiving 5.0% color died by the second week of test substance administration, and two males receiving 1.0% died during the course of the study. With the exception of five animals which died accidentally, all other animals survived the duration of the study. Based on the results obtained with respect to hepatotoxicity and body weight changes along with the histopathological data, the No observed adverse effect level (NOAEL) was considered to be 166.66 mg/kg for test chemical as no significant effects were observed at this dose .