Hesperidin

Endpoint summary

Description of key information

LD50 (rat, oral): > 2000 mg/kg bw
LC50 (rat, inhalative dust): > 5.04 g/m3

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to OECD protocol without significant deviations

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Female Wistar (RccHan™:WIST) strain rats were supplied by Harlan Laboratories UK Ltd., Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non-pregnant. After an acclimatisation period of at least five days the animals were selected at random and given a number unique within the study by indelible ink-marking on the tail and a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The bodyweight variation did not exceed ±20% of the bodyweight of the initially dosed animal.
The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

For the purpose of the study the test item was freshly prepared, as required, as a solution in dimethyl sulphoxide. Dimethyl sulphoxide was used because the test item did not dissolve in distilled water or arachis oil BP. The test item was formulated within two hours of being applied to the test system.

Doses:

Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose with one animal. In the absence of mortality at a dose level of 2000 mg/kg, an additional group of 4 animals was treated with the same dose. A total of five animals were therefore treated at a dose level of 2000 mg/kg in the study.

Control animals:

no

Details on study design:

All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing. Clinical observations were made ½, 1, 2, and 4 hours after dosing and subsequently once daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14. At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Statistics:

no statistics was applied due to lack of effects

Preliminary study:

One animal was exposed to 2000 mg/kg body weight showing no mortality.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was seen in the study

Clinical signs:

other: Hunched posture was noted during the day of dosing in four animals. No signs of systemic toxicity were noted in one animal.

Gross pathology:

No abnormalities were noted at necropsy.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral median lethal dose (LD 50 ) of the test item hesperidin in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Executive summary:

The acute oral median lethal dose (LD 50 ) of the test item hesperidin in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight. No mortality was observed in this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

6.4.1989 - 26.4.1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well documented guidline study prior to GLP

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

GLP compliance:

no

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

inhalation: dust

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

A 550 L inhalation chamber was used and test substance concentration, oxygen content and temperature were controlled. Compressed air was used and filtered prior to use for suspension of dust into the chamber. Concentration during exposure was measured as 5.05 ±0.83 g/m3 and median diameter of particles was 8.6 µm. Oxygen concentration was 21% vol and temperature was 25.0-25.3 °C

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5.04 g/m3 was used as limit concentration for exposure, measured as 5.05 ±0.83 g/m3

No. of animals per sex per dose:

10 rats were used (5 males and 5 females)

Control animals:

yes

Details on study design:

Exposure time to test atmosphere was 4 hours and 15 minutes. Thereafter, animals were observed for 14 days and body weight was recorded daily prior to necropsy.

Statistics:

no data

Preliminary study:

No mortality was observed, neither in the group of animals exposed to 5.04 g/m3 diosmin nor in the control group. Weight gain of animals during 14 day observation period was normal and did not show abnormalities.

Sex:

male

Dose descriptor:

LC0

Effect level:

> 5.04 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Sex:

female

Dose descriptor:

LC0

Effect level:

> 5.04 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Sex:

male/female

Dose descriptor:

LC0

Effect level:

> 5.04 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.04 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

no mortality was observed throughout the experiment

Clinical signs:

other: clinical signs during exposure and observation period were normal (equivalent in test group and control group)

Body weight:

body weigth gain was normal for all animals used in the test.

Gross pathology:

no macroscopic abnormalities were noted following autopsy of the test animals following the experiment

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In a limit test for acute inhalative toxicity to rats using whole body dust exposure no mortality was observed at 5.04 g/m3 and no clinical signs were noted. Following the experiment no macroscopical abnormalities were noted during autopsy. Thus the substance is considered practically nontoxic by inhalative exposire to rats.

Executive summary:

The tested Flanoides 90% diosmine contains 90% diosmin, a structurally very similar substance to hesperidin and 10% hesperidin.

During the investigation for acute inhalative toxicity to rats following dust exposure no mortality and no clinical signs were noted in any of the animals following exposure to the limit concentration of 5.04 g/m3, exposed to for 4 hours. In addition autopsy did not reveal any abnormalities in the test group in any organs. Thus the tested material is considered practically nontoxic and it can be reasonably assumed that neither diosmin nor hesperidin poses any risk by inhalative exposure to rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral toxicity:

The acute oral median lethal dose (LD 50 ) of the test item hesperidin in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight. No mortality was observed in this study.

Acute inhalative toxicity:

The tested Flanoides 90% diosmine contains 90% diosmin, a structurally very similar substance to hesperidin, and 10% hesperidin (see Read across justification in section 13).

During the investigation for acute inhalative toxicity to rats following dust exposure no mortality and no clinical signs were noted in any of the animals following exposure to the limit concentration of 5.04 g/m3, exposed to 4 hours. In addition autopsy did not reveal any abnormalities in the test group in any organs. Thus the tested material is considered practically nontoxic and it can be reasonably assumed that neither diosmin nor hesperidin poses any risk by inhalative exposure to rats.

Acute dermal toxicity:

No acute toxicity studies using dermal application are available.Although one study of a structurally very similar substance was identified showing no effects at 2000 mg/kg bw upon dermal application, the owner of this study did not grant access to this study and thus we are not allowed to use this result in this dossier.

Justification for selection of acute toxicity – oral endpoint
GLP compliant guideline study without deviations. The LC50 found was > 2000 mg/kg bw.

Justification for selection of acute toxicity – inhalation endpoint
GLP compliant guideline study without deviations. The LC0 found for diosmin (suurogate, containing 10% hesperidin) was found to be > 5.04 mg/L air.

Justification for classification or non-classification

The substance does not fulfill any of the criteria for classification according to CLP (Regulation (EC) No 1272/2008) respectively DSD (Directive 67/548/EEC). This applies to all routes of exposure.