Capsicum annuum, ext.

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

publication

Title:

A 13-week subchronic toxicity study of paprika color in F344 rats

Author:

Kanki K

Year:

2003

Bibliographic source:

Food and Chemical Toxicology 41 (2003) 1337–1343

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Capsicum annuum, ext.

EC Number:

283-403-6

EC Name:

Capsicum annuum, ext.

Cas Number:

84625-29-6

Molecular formula:

Not available since an UVBC substance.

IUPAC Name:

Capsicum annuum, ext.

Test material form:

liquid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: The stability of the test item contained in diet was assessed by San-Ei gen F.F.I., Inc. with test preparations of 5, 1, 0.2, and 0.04% paprika color (Lot No. 000927) in Oriental MF powder diet stored for 7, 14, 28, 56, and 84 days under the conditions of room lighting (approx 600 lx) at room temperature or in the dark at 5ºC. The color value of the chemical remained over 80% in the diet preparations for 28 days with the lighted condition and over 90% for 84 days in the dark. Therefore, diets were newly prepared every 2 weeks and preserved in the dark condition at 4ºC prior to use.

Test animals

Species:

rat

Strain:

Fischer 344/DuCrj

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan (Konagawa, Japan)
- Age at study initiation: 5 weeks
- Weight at study initiation: Males: 100 g; females: 80 g
- Housing: The animals were housed in a room with a barrier system, in plastic cages (five rats/cage) on soft chip bedding (Sankyo Labo-service, Tokyo, Japan). Throughout the experiment, chips were renewed every 3 days.
- Diet (e.g. ad libitum): CRF-1 powder diet (Oriental Yeast Co., Ltd., Tokyo).
- Water (e.g. ad libitum): tap water ad libitum.
- Acclimation period: 1 week.

ENVIRONMENTAL CONDITIONS
- Temperature (ºC): 24 ± 1ºC
- Humidity (%): 55 ± 5 %
- Air changes (per hour): 18 / hour
- Photoperiod (hours dark / hours light): 12 hours dark / 12 hours light

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): every 2 weeks
- Mixing appropriate amounts with (Type of food): CRF-1 powder diet (Oriental Yeast Co., Ltd., Tokyo).
- Storage temperature of food: 4ºC

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 other: % in diet

Doses / concentrations 2

Dose / conc.:

0.62 other: % in diet

Remarks:

Equivalent to 385.40 (males) and 419.00 (females) mg/kg bw/day.

Doses / concentrations 3

Dose / conc.:

1.25 other: % in diet

Remarks:

Equivalent to 749.90 (males) and 803.60 (females) mg/kg bw/day.

Doses / concentrations 4

Dose / conc.:

2.5 other: % in diet

Remarks:

Equivalent to 1506.00 (males) and 1641.10 (females) mg/kg bw/day.

Doses / concentrations 5

Dose / conc.:

5 other: % in diet

Remarks:

Equivalent to 2948.40 (males) and 3197.40 (females) mg/kg bw/day.

No. of animals per sex per dose:

10

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale:
- Fasting period before blood sampling for clinical biochemistry: 16 h

Positive control:

No

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: once a week

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: prior to necropsy
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all
- Parameters checked: white blood cell count (WBC), red blood cell count (RBC), hemoglobin concentration (Hb), hematocrit (Ht), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and platelets (PLT).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: prior to necropsy
- Animals fasted: Yes
- How many animals: all
- Parameters checked: total protein (TP), albumin/globulin ratio (A/G), albumin (Alb), total bilirubin (T.Bil), triglyceride (TG), total cholesterol (T.Cho), blood urea nitrogen (BUN), creatinine (CRN), aspartate aminotransferase (AsT), alanine aminotransferase (AlT), g-glutamyl transaminase (g-GT), alkaline phosphatase (ALP), calcium (Ca), inorganic phosphate (P), sodium (Na), potassium (K) and chloride (Cl).

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; necropsy performed on all animals. Weights of the brain, heart, lungs, thymus, liver, spleen, adrenals, kidneys, and testes were measured.
HISTOPATHOLOGY: Yes; perfomed on all animals of both sexes from the control and the highest dose groups for brain, heart, lungs, thymus, liver, spleen, adrenals, kidneys, testes, cranium with nasal cavity, pituitary, eyeballs, Harderian glands, spinal cord, salivary glands, stomach, small and large intestine, pancreas, urinary bladder, skin, mammary gland, mesenteric lymph nodes, trachea, esophagus, thyroid gland, tongue, skeletal muscle, ischiatic nerve, epididymis, seminal vesicles, prostate gland, uterus, ovary and vagina. The right testis of five animals from the control and the highest dose groups were fixed in Bouin’s solution.

Statistics:

The data for body and relative organ weights, hematology and serum biochemistry were analyzed statistically. The Bartlett’s test was applied to test homogeneity of variance between groups. If no significant heterogeneity was detected, one-way analysis of variance was applied. If significant heterogeneity of variance wasdetected, a Kruskal–Wallis’s test was conducted. If parameters were found to be significant between groups, a Bonferroni/Dunn’s multiple comparison was conducted.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

No remarkable changes in general appearance were observed.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no differences between control and chemical-treated groups, with very little inter-group variation throughout the administration period in either sex.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

There was no suppression of food intake during the experiment, and average intakes per day were approximately 14 g in male rats and 9 g in female rats.
Total intakes of paprika color (g/kg bw) over 13 weeks were 265.4 g in 5%, 135.5 g in 2.5%, 67.5 g in 1.25% and 34.7 g in 0.62% of males, 287.7 g in 5%, 147.7 g in 2.5%, 72.3 g in 1.25% and 37.7 g in 0.62% of females. Thus, clear dose dependency of chemical intake was observed for both sexes.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not specified

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Significant WBC increase was observed in 0.62 and 2.5% males. On white blood cell differential counts, eosinophilic leukocytes were significantly decreased in 2.5% males. MCH and MCHC were significantly decreased in 2.5% females.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Significant increase of T. Cho in a dose related manner was observed in 1.25, 2.5, and 5% males. CRN was significantly decreased in 1.25% males and Ca was also significantly decreased in 2.5 and 5% males. In contrast, significant increases of Na and Cl were observed in 5% males. In females, significant increases were observed in T. Cho in the 2.5 and 5% groups, Ca in all chemical-treated groups and AlT in the 0.62 and 5% groups, respectively.
The preparation used in this study mainly consists of fatty acids such as capsanthin and b-carotene at the level of 95.9%, which might have caused the dose-dependent increase of serum T. Cho.

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

There were no significant differences among the experimental groups in either sex.

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

The histopathological examination showed slight inflammatory cell infiltration in the cardiac muscle of control and 5% males without inter-group differences. Although vacuolation of liver cells was frequently observed in control and 5% males, there was no clear
difference in the severity of lesions between groups. No toxicological changes were found in the testis.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

2 948.4 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: Little or no significant toxicity was found up to 5% of test item in the diet.

2

Key result

Dose descriptor:

NOAEL

Effect level:

3 197.4 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: Little or no significant toxicity was found up to 5% of test item in the diet.

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

The NOAEL of the test substance was concluded to be 5% in the diet (0.67 g/rat/day or 2948.4 mg/kg bw/day for male rats and 0.43 g/rat/day or 3197.4 mg/kg bw/day for female rats).

Executive summary:

In a 13-week subchronic toxicity study performed similarly to OECD Guideline 408, F344 rats divided in 5 groups each consisting of 10 males and 10 females were fed powder diet containing paprika color at dose levels of 0 (basal diet), 0.62, 1.25, 2.5 and 5% (maximum). Clinical signs and general appearance were observed once a day and body weights and food intakes were measured once a week. An autopsy was performed at the end of the experiment and histopathological examinations were carried out on the 5% and basal diet (control) groups for both sexes. During the experiment, there were no remarkable changes in general appearance and no deaths occurred in any experimental group. Although serum total cholesterol was dose-dependently increased in both sexes, no related histopathological changes were observed in the liver. Slight inflammatory cell infiltration in the myocardium and vacuolation of hepatocytes were noted in both control and paprika color treated animals, but there were no clear differences between groups. In conclusion, paprika color even at 5% in the diet (0.67 g/rat/day or 2948.4 mg/kg bw/day for male rats and 0.43 g/rat/day or 3197.4 mg/kg bw/day for female rats) did not cause remarkable adverse effects in F344 rats.