Hesperidin

Endpoint summary

Description of key information

A NOAEL of 750 mg/kg bw/d was seen in a feeding study on rats with 200 days of exposure time. No substance related effects were observed in this study. This correspond with the information that hesperidin, being present in citrus fruits at concentration up to 12%, is part of daily diet of mankind.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1939

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The test is not a guideline method nor GLP compliant, but fulfills basically scientific principles.

Qualifier:

no guideline followed

Principles of method if other than guideline:

As the study had been performed in 1939, prior to any guideline being available, no guideline could be followed. However, basic scientific principles were followed incl. negative control (no treatment) and two different concentrations of test substance application.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

other: Albino

Sex:

male

Details on test animals or test system and environmental conditions:

The rats averaging 50 grams body weight were used at the start of the feeding experiment.

Route of administration:

oral: feed

Vehicle:

not specified

Details on oral exposure:

Hesperidin was mixed with the stock diet in concentrations up to one percent by weight. The animals had free access to food and water at all times.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

200 days

Frequency of treatment:

continuous

Dose / conc.:

0.062 other: percent by body weight

Dose / conc.:

1 other: percent by body weight

No. of animals per sex per dose:

six to eight rats per dose level, sex not specified

Control animals:

yes, concurrent no treatment

Details on study design:

Male albino rats averaging 50 grams body weight at the start of the feeding experiment were used, six to eight rats for each dosage level. The substance was mixed with the stock diet in concentrations up to one percent by weight.The animals had free access to food and water at all times.

Positive control:

no data

Observations and examinations performed and frequency:

Observations were made on growth curves, weights of important viscera, and macroscopic and microscopic examination of these tissues.

Sacrifice and pathology:

Histological examination of paraffin sections stained with hematoxylin-eosin

Statistics:

no data

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

During the last 50 days respiratory infection was observed throughout the laboratory colony, not related the test article affecting weight gain in this last period of experiment.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Growth curves of negative controls and dose groups fully parallel.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Description (incidence and severity):

only blood sugar values were monitored

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

During the last 50 days respiratory infection was observed throughout the laboratory colony, not related the test article.

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

The weights of the hearts, spleens, livers, adrenals, kidneys and testes were all within normal range and no macroscopical abnormalities were noted. Histopathological examinations showed no significant morphological changes.

Histopathological findings: neoplastic:

not specified

Details on results:

At the end of 200 days all the rats were killed with ether and autopsied. The weights of the hearts, spleens, livers, adrenals. kidneys, and testes were all within the normal range. All organs appeared normal range. All organs appeared normal macroscopically except the lungs of rats having respiratory infection. Histological examination of paraffin sections stained with hematoxylin-eosin showed no significant morphological changes in the livers, hearts, kidneys, spleens, adrenals. and testes of rats receiving hesperidin.

Dose descriptor:

NOAEL

Effect level:

1 other: percent by weight in food

Based on:

test mat.

Sex:

male

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

No effects were observed; 1% in diet applied to rats were converted to ca. 750 mg/kg/day calculated for rats with a mean body weight of ca. 250 g and ca. 20 g/day of food consumption.

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

No effects were observed; 1% in diet applied to rats were converted to ca. 750 mg/kg/day calculated for rats with a mean body weight of ca. 250 g and ca. 20 g/day of food consumption.

Critical effects observed:

not specified

Conclusions:

There was no difference between control rats and those receiving hesperidin in the test, giving a negative result. Thus, the NOAEL can be set to 1% (ca. 750 mg) by weight per day in food in this study.

Executive summary:

The continued feeding to healthy albino rats of hesperidin in a standard diet for a period of 200 days, in concentrations as high as one percent of the diet by weight, gave no evidences of cumulative injury as judged by growth curves, weights of important viscera, and macroscopic and microscopic examination of these tissues. There was no difference between control rats and those receiving hesperidin in the test, giving a negative result. Thus, the NOAEL can be set to 1% (ca. 750 mg) by weight per day in food in this study. This value is consistent with another NOAEL value on Neohesperidin dihydrochalcone (a surrogate to hesperidin) in the study of B.A.R. Lina, et al. (1990) and taking into account that rats of 50 g weight were used at the start of the experiment.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

750 mg/kg bw/day

Study duration:

subchronic

Species:

rat

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

Additional information

The study performed by Wilson et al. in 1939 investigated the effect of hesperidin at doses up to 1% by weight in food over a period of 200 days. Despite an lung inflammatory effect observed as of day 150, that occured within the entire laboratory colony and therefore is not treatment related, no adverse effects were noted. Therefore, 1% hesperidin in food by weight is considered as NOAEL which can be converted to 750 mg/kg bw/d (assuming an average weight of rats of ca. 250 g and ca. 20g food consumption per day). This result is consistent with the NOAEL value reported by B.A.R. Lina, et al. (1990) for neohesperidin dihydrochalcone from a subchronic oral feeding study. This finding is additionally supported by sub-chronic studies on Methyl hesperidin by Kawabe et al. Both studies do not show adverse effects at doses up to 1% by weight in diet. Only at 5% by weight in food slight effects were noted.

The suitability for methyl hesperidin and neohesperidin dihydrochalcone is discussed in the read across justification in section 13 and both substance are structurally very similar to hesperidin and do share common metabolistic pathways.

In 2002 Ohtsuki et al. reported an investigation to study the curing effects of hesperidin and glucosyl hesperidin (a solubilized form of hesperidin) on spontaneously hypersensitive rats. At a dose of 30 mg/kg bw/d, administered over a period of 25 weeks, effects on blood pressure reduction were noted to these hypersensitive rats, indicating a positive health effect - no other effects were described. However, for this study only an abstract was available for assessment and the reliability of this study could not be assessed (Klimisch 4).

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Although a very old study it was performed with high dose (1% in food over 200 days of exposure) and is well documented.

Justification for classification or non-classification

A 200 days feeding study on rats revealed no signs of systemic toxicity or negative effects in a dose relevant for classification. Hence, it can be concluded that the substance does not meet the criteria for classification and labeling for repeated dose toxicity (STOT), as set out in Regulation (EC) NO. 1272/2008 and in Directive 67/548/EEC respectively, as indicated by study results on hesperidin. Supporting studies on Methyl hesperidin and Neohesperidin dihydrochalcone (both structurally very similar sharing common metabolistic pathways) support this finding.