Capsicum annuum, ext.

Endpoint summary

Description of key information

Key study: Test method similar to OECD TG 408. In a 13-week subchronic study, paprika color showed little or no significant toxicity even with 5% supplementation in the diet. Thus, the NOAEL 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).

Key study: Test method similar to OECD TG 452. In a 52-week chronic study, the NOEL was estimated to be 2.5% in the diet (1253 mg/kg bw/day) and the NOAEL was determined to be 5% in the diet (2388 mg/kg bw/day) for male rats, and for females, the NOEL was concluded to be 5% in the diet (2826 mg/kg bw/day).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

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

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

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.

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

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

Dose / conc.:

0 other: % in diet

Dose / conc.:

0.62 other: % in diet

Remarks:

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

Dose / conc.:

1.25 other: % in diet

Remarks:

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

Dose / conc.:

2.5 other: % in diet

Remarks:

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

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

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.

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

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.

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.

Key result

Critical effects observed:

no

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.

Reference 2

Endpoint:

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

Qualifier:

according to guideline

Guideline:

other: Guidelines for Designation of Food Additives, and for Revision of Standard for Use of Food Additives

Version / remarks:

MHLW (Ministry of Health, Labor and Welfare of Japan), 1996b. Guidelines for Designation of Food Additives, and for Revision of Standard for Use of Food Additives. Article No. 29 of the Life and Sanitation Bureau.

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

GLP compliance:

not specified

Limit test:

no

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.

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 (Kanagawa, Japan)
- Age at study initiation: 6 weeks
- Weight at study initiation: Males: 120 g; females: 100 g
- Housing: The animals were housed in a room with a barrier system, in plastic cages (three or four rats/cage) on soft chip bedding (Sankyo Labo-service, Tokyo, Japan). Throughout the experiment, chips were renewed every 3 or 4 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: 2 weeks.

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

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:

52 weeks

Frequency of treatment:

Daily

Dose / conc.:

0 other: % in diet

Dose / conc.:

0.62 other: % in diet

Remarks:

Equivalent to 320 (males) and 360 (females) mg/kg bw/day.

Dose / conc.:

1.25 other: % in diet

Remarks:

Equivalent to 667 (males) and 731 (females) mg/kg bw/day.

Dose / conc.:

2.5 other: % in diet

Remarks:

Equivalent to 1253 (males) and 1401 (females) mg/kg bw/day.

Dose / conc.:

5 other: % in diet

Remarks:

Equivalent to 2388 (males) and 2826 (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: overninght

Positive control:

No

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: every week until week 5 and every 5 weeks thereafter.

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: red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLT) and white blood cell count (WBC).

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), alkaline phosphatase (ALP), gamma-glutamyl aminotransferase (gamma-GTP), calcium (Ca), inorganic phosphorus (P), sodium (Na), chloride (Cl) and potassium (K).

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; necropsy performed on all animals. Weights of the brain, lungs, heart, spleen, liver, 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, lungs, heart, spleen, liver, adrenals, kidneys, testes, cranium with nasal cavity, pituitary, eyeballs, Harderian glands, spinal cord, salivary glands, stomach, small and large intestine, caecum, pancreas, urinary bladder, skin, mammary gland, lymph nodes, sternum, trachea, esophagus, thyroid gland, tongue, femoral muscle and bone, trigeminal and ischiatic nerve, epididymis, seminal vesicles, prostate gland, coagulating gland, uterus, ovary and vagina.

Statistics:

The data for body weights, food consumption, hematology, serum biochemistry and organ weights were analyzed statistically. The Bartlett’s test was applied to test homogeneity of variance between groups. When the data were homogeneous, one-way analysis of variance (ANOVA) was applied. In the heterogeneous cases, the Kruskal–Wallis test was applied. When statistically significant differences were indicated, the Dunnett’s multiple test was employed for comparison between control and treated groups. Final survival rates and incidences of tumors and histopathological findings were compared with the Fisher’s exact probability test.

Clinical signs:

no effects observed

Description (incidence and severity):

No remarkable changes in general appearance were observed.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

One 1.25% male died due to accident, but no other deaths were observed in either sex throughout the treatment period.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

During the treatment period, significant (p < 0.05) suppression of body weight gain was observed in week 19 and 24 in 0.62% males and in week 2 in 5% females, and significant increase was observed from weeks 2 to 4 in 1.25% males and in week 3 in 2.5% males. In the final body weights, no treatment-dependent variation was observed in either sex.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No test substance-related change was found.
Intake of paprika color for 0.62%, 1.25%, 2.5% and 5% groups were estimated to be 320, 677, 1253 and 2388 mg/kg bw/day for males, and 360, 731, 1401 and 2826 mg/kg bw/day for females, respectively. Thus, test substance showed a good correlation with the expected doses.

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):

No dose-related changes in hematological parameters were found, though a significant increase of MCV was observed in 1.25% and 5% females.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

A significant increase of ALP was observed in 0.62% and 5% females and all treatment groups other than 5% in males. Na and Cl were increased in 5% males, and TP and Ca in 2.5% and 5% females were decreased with significance, but no dose-related alteration was evident.

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Significant increase in absolute weights of the kidneys in 1.25% males, and significant decrease in relative weights of the spleen in 5% males were noted. These effects were considered of little toxicological significance because of no dose-dependent manner.

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Several non-neoplastic lesions, such as focal necrosis and bile duct hyperplasia in the male liver and cystic dilatation in the female pituitary, were noted in control and 5% groups, but there were no significant differences in their incidences between the groups in either sex, except for an increased incidence of vacuolation of hepatocytes in 5% males.

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

As neoplastic lesions, a pheochromocytoma in an adrenal of a control male and an adenoma in the anterior pituitary in 5% male were observed.

Other effects:

no effects observed

Key result

Dose descriptor:

NOEL

Effect level:

1 253 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

histopathology: non-neoplastic

Key result

Dose descriptor:

NOAEL

Effect level:

2 388 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: neoplastic

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.

Key result

Dose descriptor:

NOEL

Effect level:

2 826 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: neoplastic

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.

Key result

Critical effects observed:

no

Conclusions:

The NOAEL of the test substance was concluded to be 5% in the diet (2388 mg/kg bw/day for male rats and 2826 mg/kg bw/day for female rats).

Executive summary:

In a chronic toxicity study performed similarly to OECD Guideline 452, 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) for 52 weeks. Clinical signs and general appearance were observed once a day and body weights and food consumption were measured every week until week 5 and every 5 weeks thereafter. 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. Treatment with paprika color caused a significant increase in incidence of hepatocellular vacuolation in 5% males, but no toxicological effects were found with reference to survival rates, body weights, hematological or serum biochemical parameters and organ weights at any dose level in either sex. In conclusion, based on slight histopathological changes observed in 5% male livers, the NOEL was estimated to be 2.5% in the diet (1253 mg/kg bw/day) and the NOAEL was determined to be 5% in the diet (2388 mg/kg bw/day) for male rats, and for females, the NOEL was concluded to be 5% in the diet (2826 mg/kg bw/day).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

2 388 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

2 key studies are available (subchronic and cronic) with a Klimisch score of 2.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the available data, the substance is not classified for specific target organ toxicity by repeated exposure (STOT-RE) according to CLP Regulation (EC) no 1272/2008.