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Carcinogenicity

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Description of key information

The Carcinogenic Low Observed Adverse Effect Level (LOAEL) of the test compound D&C Yellow No. 11 is found to be 500 ppm (25 mg/Kg bw) in male and female F344/N rats.

Key value for chemical safety assessment

Carcinogenicity: via oral route

Link to relevant study records
Reference
Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data available from study report.
Qualifier:
according to
Guideline:
other: Data is from Journal with permission
Principles of method if other than guideline:
The toxicity of intake of 0,500, 1,700, or 5,000 ppm D & C Yellow in rat was assessed over a 105 (Males) to 106 weeks (Females) (2 Year) experiment.
Data is from NATIONAL TOXICOLOGY PROGRAM (NTP) report no.463
GLP compliance:
not specified
Species:
rat
Strain:
other: F344/N
Sex:
male/female
Details on test animals and environmental conditions:
Details on test animal and environmental conditions:
TEST ANIMALS
- Source: Bred in-house
- Age at study initiation: 28 days at weaning
- Weight at study initiation: No data available
- Fasting period before study: No data available
- Housing: 5/cage in solid bottom polycarbonate cages with Sani chips bedding
- Diet (e.g. ad libitum): NIH-07 open formula mash (Zeigler Brothers, Inc., Gardners, PA), available ad libitum
- Water (e.g. ad libitum): Tap water (Birmingham municipal supply) via automatic watering (Edstrom Industries, Inc., Waterford, WI), available ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0-25.6Oc
- Humidity (%):23.3% to 81.2%
- Air changes (per hr): minimum of 10 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours/day
Route of administration:
oral: feed
Type of inhalation exposure (if applicable):
not specified
Vehicle:
not specified
Details on exposure:
Rate of preparation of diet (frequency):diet was prepared every two weeks
- Mixing appropriate amounts with: The dose formulations were prepared every 2 weeks by mixing D&C Yellow No. 11 with feed . A D&C Yellow No. 11/feed premix was made by hand and then blended with feed in a Patterson-Kelly twin-shell blender for 15 minutes with the intensifier bar on for the first 5 minutes.
- Storage temperature of food: During the studies, dose formulations were stored in double-thickness plastic bags in rigid plastic containers at room temperature protected from light for up to 3 weeks.
VEHICLE
- Concentration in vehicle: 0, 500, 1,700, or 5,000 ppm
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Yes, Periodic analyses of the dose formulations of D&C Yellow No. 11 were conducted at the study laboratory using visible spectrometry. Dose formulations were analyzed approximately every 8 to 10 weeks
Duration of treatment / exposure:
Duration of exposure:
Males: 105 weeks
Females: 106 weeks

Duration of study: 2 years
Frequency of treatment:
Daily
Post exposure period:
No data available
Remarks:
Doses / Concentrations:
0,500, 1,700, or 5,000 ppm; Males: 0, 25, 85 and 250 mg/Kg bw; Females: 0, 25, 100 and 280 mg/Kg bw
Basis:
nominal in diet
No. of animals per sex per dose:
60/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
Details on study design
- Dose selection rationale: The results of the 13-week rat study were used to select doses for the current F344/N rat study
-Rationale for animal assignment:
The results of the 13-week rat study were used to select doses of 500, 1,700, and 5,000 ppm for the current F344/N rat study.

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice weekly
- Cage side observations checked in table were included: Yes

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: recorded initially, weekly for 13 weeks, monthly thereafter, and at the end of the studies

DERMAL IRRITATION : No

BODY WEIGHT: Yes
- Time schedule for examinations: recorded initially, weekly for 13 weeks, monthly thereafter, and at the end of the studies.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes / No data available

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

WATER CONSUMPTION AND COMPOUND INTAKE: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood was collected from the retroorbital sinus of all 12-month interim evaluation rats.
- Anaesthetic used for blood collection: CO2/O2 mixture & blood was drawn from the retroorbital sinus.
Animals fasted: No data
- How many animals: 10 male and 10 female rats.
- Parameters checked in table were examined- hematocrit; hemoglobin; erythrocyte, reticulocyte, and nucleated erythrocyte counts; mean cell volume; mean cell hemoglobin; mean cell hemoglobin concentration; platelet counts; and total leukocyte counts and differentials

CLINICAL CHEMISTRY: None
- Time schedule for collection of blood:
Blood was collected from the retroorbital sinus of all 12-month interim evaluation rats.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Organ weight: At the 12-month interim evaluation, the liver and right kidney were weighed

Sacrifice and pathology:
GROSS PATHOLOGY: Yes, At necropsy, all organs and tissues were examined for grossly visible lesions

HISTOPATHOLOGY: Yes
In addition to gross lesions and tissue masses, the tissues examined included: adrenal gland, brain, clitoral gland, esophagus, femur with marrow and epiphysis, heart and aorta, large intestine (cecum, colon, and rectum), small intestine (duodenum, jejunum, and ileum), kidneys, liver, lungs and mainstem bronchi, lymph nodes (mandibular and mesenteric), mammary gland, nose, ovaries, pancreas, parathyroid gland, pituitary gland, preputial gland, prostate gland, salivary gland, skin, spleen, stomach (forestomach and glandular stomach), testes, thymus, thyroid gland, trachea, urinary bladder, and uterus.
Other examinations:
Necropsy performed on all animals. Organs weighed at the 12-month interim evaluation were the liver and right kidney.
Statistics:
Survival Analyses: The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1958) and is presented in the form of graphs.

Statistical analyses for possible dose-related effects on survival used Cox’ s (1972) method for testing two groups for equality and Tarone’s (1975) life table test to identify dose-related trends.


Calculation of Incidence: The Kaplan-Meier estimate of the neoplasm incidence that would have been observed at the end of the study in the absence of mortality from all other competing risks.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
All animals were observed twice daily. Clinical findings and body weights were recorded at least once a week for the first 13 weeks and every 4 week s thereafter.
Mortality:
mortality observed, treatment-related
Description (incidence):
All animals were observed twice daily. Clinical findings and body weights were recorded at least once a week for the first 13 weeks and every 4 week s thereafter.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Mean body weights of 1,700 and 5,000 ppm males and females were generally lower than those of the controls throughout the study. Final mean body weights of males were 95% (500 ppm), 93% (1,700 ppm), and 85% (5,000 ppm) that of the controls, and those of
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Feed consumption by exposed groups was similar to that by the controls
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
not specified
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
not specified
Details on results:
Body weight and weight gain:
Mean body weights of 1,700 and 5,000 ppm males and females were generally lower than those of the controls throughout the study.
Final mean body weights of males were 95% (500 ppm), 93% (1,700 ppm), and 85% (5,000 ppm) that of the controls, and those of females were 99%, 95%, and 94% that of the controls.

Haematology:
There was evidence of minimal anemia in exposed males; this anemia was characterized by decreased hematocrit values, hemoglobin concentrations, and erythrocyte counts. There were no differences in the mean cell volume or mean cell hemoglobin concentration in exposed rats, to indicate that erythrocytes were normocytic and normochromic. There were no increases in reticulocyte counts to indicate a bone marrow response to the anemia. Therefore, the minimal anemia was characterized as normocytic , normochromic, and nonresponsive. Normocytic , normochromic, nonresponsive anemias have been related to selective suppression of erythropoiesis in a variety of disorders and may be due to decreased erythropoietin elaboration, bone marrow suppression, or defective iron metabolism.
There were no biologically or statistically significant differences in hematology parameters between control and exposed females.

Histopathology: non-neoplastic:
In Males-
Liver: clear cell focus (9/50, 15/51, 15/51, 18/54);
mixed cell focus (1/50, 10/51, 9/51, 10/54); bile duct pigmentation (0/50, 38/51, 51/51, 54/54) hepatocyte cytologic alterations (0/50, 20/51, 44/51,42/54); hepatocyte pigmentation (0/50, 22/51, 45/51, 51/54); Kupffer cell pigmentation (7/50, 15/51, 23/51, 26/54)
Kidney: renal tubule hyperplasia evaluation – 0/50, 0/51, 4/51, 3/54; extended evaluation – 0/50, 2/51, 9/51, 2/54; standard and extended evaluations combined – 0/50, 2/51, 13/51, 4/54); renal tubule pigmentation (18/50, 43/51, 4/54); renal tubule pigmentation (18/50, 43/51, 47/51, 54/54); transitional epithelial hyperplasia (11/50, 23/51, 29/51, 34/54); severity of nephropathy (2.3, 2.8, 3.2, 3.0)
In Females-
Liver: clear cell focus (10/50, 18/51, 29/50 30/51); bile duct pigmentation (0/50, 46/51, 49/50, 50/51); hepatocyte cytologic alterations pigmentation (0/50, 34/51, 44/50, 50/51) Kupffer cell pigmentation (9/50, 11/51, 16/50, 32/51).
Kidney: renal tubule pigmentation (10/50, 48/51,
50/50, 51/51); transitional epithelial hyperplasia
(2/50, 6/51, 10/50, 3/51); severity of nephropathy
(1.4, 1.7, 1.8, 2.1)

Histopathology: neoplastic:
In males-
Liver: Hepatocellular adenoma (1/50, 2/51, 1/51,7/54)
Kidney: renal tubule adenoma (standard
standard evaluation – 0/50, 0/51, 0/51, 2/54; extended evaluation – 0/50, 2/51, 4/51, 2/54; standard and extended evaluations combined – 0/50, 2/51, 4/51, 4/54); renal tubule adenoma or carcinoma (standard and extended evaluations combined – 0/50, 2/51, 5/51, 4/54) Oral cavity: squamous cell papilloma (1/50, 1/51, 2/51, 4/54); squamous cell carcinoma (0/50, 1/51, 1/51, 1/54); squamous cell papilloma or squamous cell carcinoma (1/50, 2/51, 3/51, 5/54)
In females
Liver: hepatocellular adenoma or carcinoma (0/50, 2/51, 5/50, 5/51).Complete histopathology was performed on all rats
Dose descriptor:
LOAEL
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Neoplastic and non neoplastic effects observed
Remarks on result:
other: Effect type: carcinogenicity (migrated information)

Clinical sign and mortality

 

0 ppm

500 ppm

1700 ppm

5000ppm

Male

 

Total animals

60

60

60

60

12 month interim evaluation

10

9

9

6

Moribund

29

29

41

49

Natural death

2

2

1

3

Females

 

Total animals

60

60

60

60

12 month interim evaluation

10

9

10

9

Moribund

25

23

12

25

Natural death

3

2

1

3

 

Clinical Signs: Chemical-related clinical findings included yellow discoloration of the entire body in all exposed males and females from day 1 and head swelling and edema in 1,700 and 5,000 ppm males. One 1,700 ppm male and five 5,000 ppm males were killed moribund between weeks 49 and 81; these deaths were attributed to extensive edema.

Organ Weights:

Yes

* Significantly different (P 0.05) from the control group by Williams’ or Dunnett’s test ** P 0.01.

a Organ weights (absolute weights) and body weights are given in grams; organweight to body sweight ratios (relative weights) are given as mg organ weight/g body weight (mean ± standard error).

Male

 

0

ppm

500

ppm

1,700

ppm

5000ppm

Necropsy body wt

471 ±10

461 ± 10

446 ± 6

440 ± 13

R. Kidney

 

Absolute

1.655 ± 0.042

1.680 ± 0.046

1.589 ± 0.045

1.620 ± 0.065

Relative

3.52 ± 0.05

3.65 ± 0.06

3.56 ± 0.08

3.68 ± 0.07

Liver

 

Absolute

16.916 ± 0.599

19.417 ± 0.748**

19.599 ± 0.457**

20.480 ± 0.750**

Relative

35.89 ± 0.75

42.03 ± 0.94**

43.95 ± 1.12**

46.49 ± 0.75**

Female

 

0

ppm

500

ppm

1,700

ppm

5000

ppm

Necropsy body wt

267 ± 8

262 ± 5

257 ± 2

252 ± 6

R.Kidney

 

 

 

 

 

Absolute

0.963 ± 0.021

0.912 ± 0.020

0.920 ± 0.027

0.930 ± 0.022

Relative

3.62 ± 0.08

3.48 ± 0.05

3.58 ± 0.09

3.69 ± 0.04

Liver

 

Absolute

8.853 ± 0.204

9.656 ± 0.202*

10.566 ± 0.204**

10.780 ± 0.245**

Relative

33.21 ± 0.58

36.87 ± 0.44**

41.06 ± 0.56**

42.82 ± 0.50**

Conclusions:
The Carcinogenic Low Observed Adverse Effect Level (LOAEL) of the test compound D&C Yellow No. 11 is found to be 500 ppm (25 mg/Kg bw) in male and female F344/N rats.
Executive summary:

A 2 year toxicity study was conducted on groups of 60 male/female F344/N rats for the test compoundD&C Yellow No. 11. The chemical was administered oral through feed at dose range of 0, 500, 1700 and 5000 ppm for 105 (males) and 106 (females) weeks. 6 to 10 rats per group were evaluated at 12 months. These exposure concentrations resulted in average daily doses of approximately 25, 85 or 250 mg D&C Yellow No. 11/kg body weight to males and 25, 100 or 280 mg/kg bw to females.

Observations were made for body weights, feed consumption, clinical findings, hematology and pathological findings.

Some evidence of carcinogenic activity in male F344/N rats was observed based on increased incidences of hepatocellular adenoma, renal tubule neoplasms, and squamous cell neoplasms of the oral cavity. There was some evidence of carcinogenic activity in female F344/N rats based on increased incidences of hepatocellular neoplasms. Incidences o f uncommon squamous cell carcinoma of the oral cavity in females may have been related to chemica l treatment.

 

Exposure of rats to D&C Yellow No. 11 in feed for 2 years resulted in increased incidences of nonneoplastic liver lesions including clear cell foci, increase d basophilia and granularity in the cytoplasm of hepatocytes, and bile duct, hepatocyte, and Kupffer cell pigmentation in males and females and mixed cell foci in males. In the kidney, there were increased incidences of renal tubule pigmentation and transitional epithelial hyperplasia in males and females and renal tubule hyperplasia in males. The severity of nephropathy was increased in exposed males and females.

The Carcinogenic Low Observed Adverse Effect Level of the test compound D&C Yellow No. 11 is found to be 500 ppm (25 mg/Kg bw) in male and female F344/N rats.

According to the Publication, the test material is positive for carcinogenicity.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
250 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data from K2 publication.

Carcinogenicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Carcinogenicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The studies reviewed for Carcinogenicity from reliable sources having Klimish rating 2. 

The summary of the results are presented below

 

Sr. No.

Species

End point

Effect type

Dose Level

Effect level

1

Rat

LOAEL

Carcinogenicity

25 mg/kg bw/day (nominal)

Neoplastic and non neoplastic effects observed

2

hamster, Syrian

LOAEL

Carcinogenicity

0.1 µg/ml

A test chemical is considered positive in the SHE cell transformation assay if it causes a statistically significant (p<0.05) increase in MT (relative to the solvent control) in at least two dose groups, or if it causes a statistically significant increase in MT in one dose with a statistically significant (p<0.05)

positive dose-response trend test. If either the 24-hr or the 7-day exposure is positive, the overall SHE cell transformation assay call for a test chemical is positive

 

From above mentioned details, the target chemical D&C Yellow No. 11, it can be concluded that the substance is likely to be carcinogenic.

Justification for selection of carcinogenicity via oral route endpoint:

There was some evidence of carcinogenic activity in female F344/N rats based on increased incidences of hepatocellular neoplasms. Incidences of uncommon squamous cell carcinoma of the oral cavity in females may have been related to chemical treatment.

Justification for classification or non-classification

The avaialable study indicates that the substance D&C Yellow No. 11 is likely to be carcinogenic in nature.