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Carcinogenicity

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Description of key information

Key study. Read-across from analogue substance. Method similar to OECD 451. Study on a structural analogue to neohesperidin, investigating carcinogenicity at elevated doses such as 5% in diet, not showing any carcinogenic effects. Based on the read across approach, the target substance is not considered to be carcinogenic.

Key value for chemical safety assessment

Carcinogenicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 451 (Carcinogenicity Studies)
Deviations:
yes
Remarks:
only 2 dose levels and a control.
GLP compliance:
not specified
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Hamari Pharmaceutical Industries, Ltd (Osaka, Japan), Lot No. T24VG-177
- Purity: > 95.5%

OTHER SPECIFICS:
- Name of test material (as cited in study report): methyl hesperidin
- Molecular formula (if other than submission substance): C29H36O15
- Molecular weight (if other than submission substance): 624.5871
- Smiles notation (if other than submission s.): CC1C(O)C(O)C(O)C(OCC2C(O)C(O)C(O)C(Oc3cc4c(c(O)c3)C(=O)CC(c3ccc(OC)c(OC)c3)O4)O2)O1
- InChl (if other than submission substance): InChI=1S/C29H36O15/c1-11-22(32)24(34)26(36)28(41-11)40-10-20-23(33)25(35)27(37)29(44-20)42-13-7-14(30)21-15(31)9-17(43-19(21)8-13)12-4-5-16(38-2)18(6-12)39-3/h4-8,11,17,20,22-30,32-37H,9-10H2,1-3H3/t11-,17-,20+,22-,23+,24+,25-,26+,27+,28+,29+/m0/s1
- Structural formula attached as image file (if other than submission substance): see Fig. in attachments.
Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan Inc., Kanagawa, Japan.
- Age at study initiation: 5 wk old
- Housing: The mice were housed five per cage, under constant conditions, in plastic cages with stainless-steel grid tops and solid bases on white woodchip bedding (Beta Chip Bedding, Northeastern Products Co., NY, USA).
- Diet: Oriental MF powdered basal diet (Oriental Yeast Co., Tokyo, Japan).
- Water: ad libitum.
- Acclimation period: 1 wk

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 1
- Humidity (%): 55 ± 2
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle
Route of administration:
oral: feed
Vehicle:
corn oil
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): not specified.
- Mixing appropriate amounts with (Type of food): Oriental MF powdered basal diet (Oriental Yeast Co., Tokyo, Japan).

VEHICLE
- Justification for use and choice of vehicle (if other than water): To prevent loss of the compound as dust during diet preparation or feeding, 2% corn oil was added prior to its incorporation. Control diets also contained 2% corn oil.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
96 wk
Frequency of treatment:
once a day
Post exposure period:
8 wk
Dose / conc.:
1.25 other: % in diet
Dose / conc.:
5 other: % in diet
No. of animals per sex per dose:
50/sex/dose
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: The two dose levels used were chosen based on criteria used in previous studies: sub-chronic toxicity studies were carried out with test substance prior to the carcinogenicity experiment and the 5% level was chosen as the maximum tolerable dose for life-long treatment.
- Rationale for animal assignment (if not random): The mice were assigned to control or treatment groups according to their initial body weights using a computer-assisted classifying procedure designed to provide homogeneity of variance and equality of initial group body weight.
- To eliminate the possibility of interference by reversible changes due to the compound treatment, a recovery period of 8 wk was set.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: All mice were observed at least once a day for clinical signs.

BODY WEIGHT: Yes
- Time schedule for examinations: body weights were recorded weekly during the first 13 wk and biweekly 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: Yes
- Food and water consumption were measured over 2-day periods before each weighing and average daily and total intake of test substance were also calculated.

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

WATER CONSUMPTION AND COMPOUND INTAKE (not a drinking water study): Yes
- Time schedule for examinations: measured over 2-day periods.

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the study the mice were fasted overnight. Blood samples were taken from 10 males and 10 females of each group.
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- Parameters checked: Haematology parameters evaluated were haemoglobin concentration and haematocrit, and erythrocyte, leucocyte and platelet counts. The blood cell indices of mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration were calculated. Differential leucocyte counts and estimation of the percentage of nucleated red blood cells, anisocytosis and polychromasia were performed for each smear.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the study the mice were fasted overnight. Blood samples were taken from 10 males and 10 females of each group.
- Animals fasted: Yes
- Parameters checked: Clinical chemistry parameters determined included glutamic-pyruvic transaminase, glutamic-oxaloacetic transaminase, alkaline phosphatase, total cholesterol, total protein, albumin/globulin ratio, and urea nitrogen. These parameters were measured by autoanalyser.

URINALYSIS: Yes
- Time schedule for collection of urine: During wk 104 of the experiment fresh urine samples were obtained from 10 mice/sex/group.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked: Urinalysis included measurements of pH, protein, glucose, ketone, bilirubin, occult blood, urobilinogen and specific gravity. Specific gravity was determined with a clinical refractometer. The other measurements were made with Multistix III.

NEUROBEHAVIOURAL EXAMINATION: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- Gross and complete histology examinations were performed on all mice that died, were killed on becoming moribund, or survived until the terminal killing. During autopsy, weights were recorded for the following organs: liver, kidneys, brain, heart and testes or ovaries. Relative organ weights (organ-to-body-weight ratios) were calculated.

HISTOPATHOLOGY: Yes
- Microscopic examinations were performed after routine processing and staining with haematoxylin and eosin. The above tissue plus the following were collected for histopathological examination: salivary glands, trachea, lungs, thymus, lymph nodes, spleen, stomach, gall bladder, small intestine (duodenum, jejunum, ileum), large intestine (caecum, colon, rectum), pancreas, urinary bladder, pituitary, thyroids adrenals, prostate, seminal vesicles, uterus, mammary gland, skeletal muscle, eyes, Harderian glands, spinal cord, sciatic nerve and gross lesions.
Statistics:
All measurements were expressed as mean + SD and were analysed where appropriate by the F- and t-tests or the Welch method. The significance of differences in the incidences of non-neoplastic lesions between the different groups was evaluated by the chi-square test or by Fisher's exact probability test. All references to statistical significance in this experiment represent two-tailed P values. The analyses of differences in the survival periods between the treated and control groups were analysed by the Generalized Wilcoxon and Cox-Mantel tests.
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment-related clinical signs were observed in any of the mice throughout the study.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
The growth of male mice given 5% test substance showed a slight decrease from the 16th to the 80th week, but this difference was not observed thereafter or at the end of the study. A significant decrease, with a slight dose dependency, in the body-weight gains of female mice given test substance at levels of 1.25 and 5%, was observed from wk 24 to 96. No obvious recovery was evident during the observation period over the last 8 wk after cessation of treatment. However, since survival periods of compound-treated male and female groups were not significantly different from those of controls, body-weight changes were not associated with any effects on the survival rate.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
The average daily food consumption of the females was slightly greater than that of the males. This sex difference was reflected in both values of average daily and calculated total test article intake.
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
No biologically significant differences in urinalysis, haematology or clinical chemistry values were noted between control and treatment groups.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No biologically significant differences in urinalysis, haematology or clinical chemistry values were noted between control and treatment groups.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No biologically significant differences in urinalysis, haematology or clinical chemistry values were noted between control and treatment groups.
Behaviour (functional findings):
not specified
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
A significant decrease in absolute organ weight was observed for the kidneys of females given 5,0% (P <0.01). However, evaluation of the relative organ weight revealed increases in the brain, heart and kidneys of 1.25 and 5.0% females, and decreases were evident in the brain and testes of males given 1.25% and in the kidneys of males given 1.25 and 5%. Although these changes were statistically significant, the differences in each value between the control and experimental groups were small and no dose dependency was apparent.
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Of the observed non-neoplastic findings, thyroid cysts, cyst formation and atrophy in the ovary and cystic endometrial hyperplasia of the uterus were frequent but with no treatment-related effects. Testicular atrophy and prostatitis were also observed equally in all groups. The incidences of spontaneously occurring lymphocytic accumulation in the kidneys and urinary bladder were relatively high in the female groups, but were not clearly dose related. Chronic nephropathy was observed in the male groups, at highest levels in the controls. Thus, none of the above-mentioned findings, which included age-related degenerative alterations, were considered to be due to test substance treatment.
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
In males, predominant lesions were liver hyperplastic nodules (hepatocellular adenoma or neoplastic nodule) and hepatocellular carcinomas and adenoma and adenocarcinoma in the lung. In females, the most common tumour was malignant lymphoma/leukaemia, which infiltrated into many organs and tissues. Other minor neoplastic lesions included adenomas in the pituitary of female mice, endometrial stromal sarcomas in the uterus and adenomas in the Harderian glands of both sexes. However, these lesions occurred at comparable incidences in both control and treated mice with no apparent dose-response relationship. Thus, the observed tumours should be regarded as being of spontaneous origin and not attributable to test- treatment.
Other effects:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
5 other: % in diet
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no significant effects observed.

Table 1. Food and methyl hesperidin intake in long-term feeding study of methyl hesperidin in B6C3F1 mice.

Sex

Dose

(%)

Average food intake

(g/mouse/day)

Average test

item intake

(g/kg bw/d)

Total test

item intake

(g/kg bw/d)

Male

0

5.1

0

0

1.25

4.9

1.7

1130

5.0

5.0

7.5

5015

Female

0

5.4

0

0

1.25

5.4

2.2

1475

5.0

5.2

8.6

5802

Mice were fed diets containing 0 (control), 1.25 or 5.0% methyl hesperidin for 96 wk, after which they received control diet for 8 wk prior to authopsy.

 

Table 2. Mean absolute organ weights in mice treated with methyl hesperidin (mean ± SD).

Sex

Dose

(%)

No.

Examined

Final body weight

Mean absolute organ weight

Brain

Heart

Liver

Spleen

Kidneys

Ovaries/testes

Male

0

30

33.72 ± 4.06

0.48 ± 0.02

0.23 ± 0.04

0.23 ± 0.04

0.15 ± 0.10

1.82 ± 0.08

0.21 ± 0.02

1.25

34

35.88 ± 3.31*

0.49 ± 0.02

0.23 ± 0.06

0.23 ± 0.06

0.25 ± 0.42

1.79 ± 0.06

0.20 ± 0.02

5.0

35

34.74 ± 3.08

0.49 ±0.02

0.22 ± 0.03

0.22 ± 0.03

0.22 ± 0.29

1.70 ±0.06

0.21 ± 0.04

Female

0

36

43.19 ± 5.71

0.52 ± 0.02

0.18 ± 0.02

0.18 ± 0.02

0.33 ± 0.31

0.50 ± 0.05

0.12 ± 0.41

1.25

39

38.25 ± 5.56**

0.51 ± 0.02

0.18 ± 0.03

0.18 ± 0.03

0.32 ± 0.28

0.50 ± 0.09

0.29 ± 1.05

5.0

38

37.95 ± 5.69**

0.52 ± 0.03

0.17 ± 0.02

0.17 ± 0.02

0.32 ±0.32

0.47 ± 0.04**

0.05 ± 0.13

 

Table 3. Mean relative organ weights (% of body weight) in mice treated with methyl hesperidin (mean ± SD)

Sex

Dose

(%)

No.

Examined

Mean relative organ weight

Brain

Heart

Liver

Spleen

Kidneys

Ovaries/testes

Male

0

30

1.45 + 0.17

0.68 + 0.16

6.22 ±3.47

0.45 ±0.33

1.82 + 0.18

0.63 ± 0.08

1.25

34

1-37 ± 0.11*

0.64 ±0.15

5.78 + 2.77

0.68 + 1.14

1.79 ± 0.16**

0.57 ±0.08**

5.0

35

1.41 ±0.12

0.64 ± 0.08

6.02 ± 3.07

0.64 ± 0.85

1.70 ± 0.18**

0.59 ±0.11

Female

0

36

1.22 ± 0.17

0.42 + 0.07

4.23 ± 1.19

0.79 + 0.79

1.18 + 0.16

0.30+ 1.08

1.25

39

1.37 + 0.20**

0.49 ±0.10**

4.41 ±0.91

0.87 + 0.85

1.35 + 0.36*

0.82 + 2.98

5.0

38

1.41 +0.23**

0.46 ±0.07*

4.34 ± 1.47

0.87 + 0.89

1.27 + 0.18*

0.15 + 0.38

 

Table 4. Incidences (%) of non-neoplastic lesions in mice treated with methyl hesperidin.

Site and type of lesion

Incidence (%)

Males

Females

* Dietary level (%):

0

1.25

5.0

0

1.25

5.0

No. of mice

50

50

50

50

50

50

Thyroids

 

- Cyst formation

11 (22)

5(10)

9(18)

7(14)

5(10)

3(6)

Kidneys

 

- Lymphocytic accumulation

0

2(4)

1(2)

5(10)

2(4)

5(10)

- Chronic nephropathy

6(12)

1(2)

3(6)

0

1(2)

1(2)

Urinary bladder

 

- Lymphocytic accumulation

0

1(2)

1(2)

8(16)

9(18)

6(12)

Testes

 

- Atrophy

7(14)

5(10)

5(10)

Prostate

 

- Inflammation

5(10)

3(6)

5(10)

Ovaries

 

- Cyst formation

19(38)

22 (44)

11 (22)

- Atrophy

39 (78)

38 (76)

36 (72)

Uterus

 

- Cystic endometrial hyperplasia

46 (92)

48 (96)

45 (90)

Brain

 

- Calcification

5(10)

5(10)

3(6)

2(4)

1(2)

2(4)

Only lesions with an incidence of 6% or greater are listed.

  

Table 5. Incidences (%) of tumours in mice treated with methyl hesperidin. 

Site and type of tumour*

Incidence (%)

Males

Females

* Dietary level (%):

0

1.25

5.0

0

1.25

5.0

No. of mice

50

50

50

50

50

50

All sites

 

Malignant lymphoma/leukaemia

6(12)

13(26)

9(18)

25(50)

17(34)

19(38)

Spleen

 

Haemangioma

2(4)

 0

 0

1(2)

0

0

Haemangiosarcoma

1(2)

1(2)

 0

2(4)

0

1(2)

Bone marrow

 

Haemangioma

 0

 0

1(2)

0

0

0

Pituitary

 

Adenoma

 0

 0

 0

1(2)

6(12)

6(12)

Carcinoma

 0

 0

 0

2(4)

1(2)

0

Thyroids

 

Follicular adenoma

 0

 0

 0

1(2)

0

0

Follicular carcinoma

 0

 0

 0

0

1(2)

0

Adrenals

 

Cortical adenoma

 0

 0

1(2)

0

0

0

Pheochromocytoma

1(2)

1(2)

1(2)

1(2)

0

0

Lungs

 

Adenoma

6(12)

2(4)

3(6)

3(6)

2(4)

1(2)

Adenocarcinoma

5(10)

3(6)

1(2)

0

0

0

Forestomach

 

Papilloma

3(6)

 0

 0

1(2)

2(4)

1(2)

Squamous cell carcinoma

 0

1(2)

 0

0

0

0

Small intestine

 

Adenocarcinoma

1(2)

 0

 0

0

0

0

Liver

 

Hyperplastic nodule

17(34)

16(32)

17(34)

3(6)

1(2)

3(6)

Haemangioma

3(6)

2(4)

2(4)

2(4)

1(2)

0

Haemangiosarcoma

 0

 0

 0

1(2)

0

1(2)

Hepatocellular carcinoma

10(20)

14(28)

12(24)

1(2)

1(2)

2(4)

Kidneys

 

Adenoma

 0

 0

1(2)

0

0

1(2)

Mammary gland

 

Adenocarcinoma

 0

 0

 0

1(2)

0

0

Ovary

 

Cystadenoma

2(4)

1(2)

0

Teratoma

0

1(2)

1(2)

Granulosa-theca cell tumour

1(2)

1(2)

0

Uterus

 

Endometrial stromal polyp

2(4)

1(2)

0

Endometrial stromal sarcoma

2(4)

3(6)

5(10)

Leiomyoma

0

0

1(2)

Leiomyosarcoma

0

2(4)

0

Adenocarcinoma

1(2)

1(2)

1(2)

Bone

 

Osteosarcoma

 0

 0

 0

0

1(2)

0

Skin/subcutis

 

Papilloma

 0

1(2)

 0

0

0

0

Fibroma

 0

1(2)

 0

0

0

0

Fibrosarcoma

2(4)

 0

2(4)

1(2)

2(4)

0

Malignant fibrous histiocytoma

2(4)

 0

1(2)

0

1(2)

0

Harderian gland

 

Adenoma

1(2)

3(6)

4(8)

5(10)

4(8)

1(2)

Brain

 

Astrocytoma

0

 0

1(2)

 0

0

0

 

Conclusions:
The test item was found to be non-carcinogenic.
Executive summary:

A long-term carcinogenicity study of the test item was carried out in B6C3F1 mice (50 per sex per group) receiving dietary concentrations of 0, 1.25 or 5% for 96 wk, and then maintained on a basal diet for an additional 8 wk. Growth retardation during the experiment with final changes in organ weights was observed in females given the 1.25% dose of methyl hesperidin and in both sexes receiving the 5.0% treatment. However, no biologically significant effects were evident with respect to mortality or clinical signs. Furthermore, treatment with the test item did not result in any changes in haematology, clinical chemistry or urinalysis data. On the histological examination, no significant alterations of non-neoplastic and neoplastic lesions incidence was observed in treated mice. Thus, the results demonstrated that test substance lacked any carcinogenicity for B6C3F1 mice in the 96-wk feeding regimen used in this study. Therefore, the test item was found to be non carcinogenic.

Endpoint:
carcinogenicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See "attached justification".
Reason / purpose:
read-across source
Specific details on test material used for the study:
TEST MATERIAL
- Name of test material: neohesperidin
- IUPAC name: 5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4-oxo-3,4-dihydro-2H-chromen-7-yl 2-O-(6-deoxyhexopyranosyl)hexopyranoside
- Molecular formula: C28H34O15
- Molecular weight: 610.5606
- Smiles notation: COc1ccc(cc1O)C2CC(=O)c3c(O)cc(OC4OC(CO)C(O)C(O)C4OC5OC(C)C(O)C(O)C5O)cc3O2
- InChl: InChI= 1/C28H34O15/c1-10-21(33)23(35)25(37)27(39-10)43-26-24(36)22(34)19(9-29)42-28(26)40-12-6-14(31)20-15(32)8-17(41-18(20)7-12)11-3-4-16(38-2)13(30)5-11/h3-7,10,17,19,21-31,33-37H,8-9H2,1-2H3
Sex:
male/female
Analytical verification of doses or concentrations:
not specified
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not specified
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Other effects:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
4.88 other: % in diet
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: read-across from analogue substance.
Conclusions:
Based on the available data for the read-across approach, it is concluded that target substance has no carcinogenicity for B6C3F1 mice.
Executive summary:

A long-term carcinogenicity study of the analogue substance methyl hesperidin was carried out in B6C3F1 mice (50 per sex per group) receiving dietary concentrations of 0, 1.25 or 5% for 96 wk, and then maintained on a basal diet for an additional 8 wk. Growth retardation during the experiment with final changes in organ weights was observed in females given the 1.25% dose of methyl hesperidin and in both sexes receiving the 5.0% treatment. However, no biologically significant effects were evident with respect to mortality or clinical signs. Furthermore, treatment with the test item did not result in any changes in haematology, clinical chemistry or urinalysis data. On the histological examination, no significant alterations of non-neoplastic and neoplastic lesions incidence was observed in treated mice. Thus, the results demonstrated that test substance lacked any carcinogenicity for B6C3F1 mice in the 96-wk feeding regimen used in this study. As methyl hesperidin is structurally very similar to neohesperidin and they share direct metabolites such as hesperetin dihydrochalcone (see reporting format attached), the results can be considered representative for neohesperidin too. Based on the available information, the target substance is considered to be non-carcinogenic.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Study duration:
chronic
Species:
mouse
Quality of whole database:
The study has a Klimisch score of 2.

Carcinogenicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Carcinogenicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

A long-term carcinogenicity study was performed on the analogue substance methyl hesperidin in B6C3F1 mice. 50 mice per sex per group received dietary concentrations of 0, 1.25 or 5% of the test item for 96 wk, and were maintained on basal diet for an additional 8 wk. Growth retardation during the experiment with final changes in organ weights were observed in females given the 1.25% dose of methyl hesperidin and in both sexes receiving the 5.0% treatment. However, no biologically significant effects were evident with respect to mortality or clinical signs. Furthermore, treatment with methyl hesperidin did not result in any changes in haematology, clinical chemistry and urinalysis data. On histological examination, no significant alteration of non-neoplastic and neoplastic lesion incidence was observed in treated mice. Thus, the results demonstrated that methyl hesperidin lacked any carcinogenicity for B6C3F1 mice in the 96-wk feeding regimen used in this study. Based on the read-across approach, the target substance is considered to be non-carcinogenic.

Justification for classification or non-classification

Based on the available data, it is concluded that the substance is not classified for carcinogenicity in accordance with CLP Regulation (EC) No.1272/2008.