(S)-7-[[2-O-6-deoxy-α-L-mannopyranosyl)-β-D-glucopyranosyl]oxy]-2,3-dihydro-5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4H-1-benzopyran-4-one

Endpoint summary

Description of key information

Oral route: Weight of evidence:

- Read-across from analogue substance. Source: Method similar to OECD 420. Based on read-across approach, the oral LD50 in rats for the target substance was determined to be >= 4984 mg/kg bw.

- Read-across from analogue substance. Source: Method similar to OECD 420. Based on read-across approach, the oral LD50 in rats for the target substance was determined to be >= 4984 mg/kg bw.

- Read-across from analogue substance. Source: Method similar to OECD 474. Based on read-across approach, the oral LD50 in rats for the target substance was determined to be >= 4984 mg/kg bw.

Dermal route:

- Key study. Method according to OECD 402 (limit test), GLP study. No mortality or systemic clinical signs related to the administration of the test item were observed throughout the study. The test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Inhalation route:

- Data waiving (other justification): According to Regulation (EC) No. 1907/2006, Annex VIII, 8.5.3, column 2, the acute toxicity by inhalation route is not required, as both oral and dermal studies are available.

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:

From April 19th to May 3rd, 1973.

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 420 (Acute Oral Toxicity - Fixed Dose Method)

Principles of method if other than guideline:

- Procedure found in Section 191.1 (f)(1) of the Federal Hazardous Substance Act; NPI Standard Test No.6.

GLP compliance:

no

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

TEST MATERIAL
- Lot No. 6-RS-7, received April 5, 1973.
- Purity (HPLC): 98.5%
- Impurities: Arsenic < 3ppm, Heavy metals < 15ppm.
- Loss on drying: 10.0%
- Residue on ignition: 1.0%
- Name of test material (as cited in study report): neohesperidin dihydrochalcone (Neo-DHC)
- Molecular formula (if other than submission substance): C28H36O15
- Molecular weight (if other than submission substance): 612.5764
- Smiles notation (if other than submission s.): CC1C(O)C(O)C(O)C(OC2C(O)C(O)C(CO)OC2Oc2cc(O)c(C(=O)CCc3ccc(OC)c(O)c3)c(O)c2)O1
- InChl (if other than submission substance): InChI=1/C28H36O15/c1-11-21(34)23(36)25(38)27(40-11)43-26-24(37)22(35)19(10-29)42-28(26)41-13-8-16(32)20(17(33)9-13)14(30)5-3-12-4-6-18(39-2)15(31)7-12/h4,6-9,11,19,21-29,31-38H,3,5,10H2,1-2H3

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 234g (females) and 290g (males).
- Fasting period before study: overnight
- Diet: commercial laboratory animal feed ad libitum.
- Water: bottled spring water ad libitum.
- Acclimation period: 6d.

IN-LIFE DATES: From: April 19, 1973 To: May 3, 1973.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test material was used in a 12.5% gravimetric suspension in water.
- Amount of vehicle (if gavage):

MAXIMUM DOSE VOLUME APPLIED:

DOSAGE PREPARATION: the powdered neohesperidin dihydrochalcone was mixed with 8cc of distilled water and allowed to stand 15min prior to being administered to animals. Females received 1.2g and males 1.5g test item each.

Doses:

5000 mg/kg bw.

No. of animals per sex per dose:

5 per sex per dose

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for signs of pharmacologic activity and drug toxicity at 1, 3, 6 and 24h post-dosage. Observations were made daily thereafter to a total of 14d.
- Necropsy of survivors performed: yes. Animals sacrificed at the end of the 14-day observation period were subjected to complete gross necropsy.
- Other examinations performed: clinical signs, body weight, histopathology.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

>= 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

All animals were alive at the completion of the test.

Clinical signs:

other: No effects observed.

Gross pathology:

No gross changes observed.

Interpretation of results:

GHS criteria not met

Remarks:

EU criteria

Conclusions:

Based on read-across approach, the target substance was found to be non toxic, LD50 ≥ 5000mg/kg.

Executive summary:

The acute oral toxicity of neohesperidin dihydrochalcone on rats was studied by a method similar to OECD TG 420. A limit test was performed by administering a single dose of 5g/kg bw test item to 10 (5M/5F) Wistar rats, and observing the effects for 14 days (clinical signs, body weights). No effects were observed in any animal. Therefore, the test item was found to be non toxic, with an LD50 > 5000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

4 984 mg/kg bw

Quality of whole database:

The two studies used for read-across have a Klimisch score of 2.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From November 15th to November 29th, 2016.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Remarks:

(SPF Caw)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle – France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: the males were 7 weeks old and the females 8 weeks old.
- Weight at study initiation: the mean weight of male rats was 267.6 g, and the mean weight of female rats 222.0 g
- Housing: during the treatment, animals were kept in individual cages. They were solid-bottomed clear polycarbonate cages with a stainless steel mesh lid, containing dust free weed shavings (changed at least 2 times per week).
- Diet (e.g. ad libitum): foodstuff (ENVIGO 2016) ad libitum.
- Water (e.g. ad libitum): tap-water from public distribution system ad libitum.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

semiocclusive

Vehicle:

DMSO

Details on dermal exposure:

TEST SITE
- % coverage: at least 10%
- Type of wrap if used: non occlusive porous gauze dressing (50 mm x 50 mm non woven swab of 4-layer patch from MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic microporeTM adhesive tape from 3M)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): rinsing with distilled water.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): 0.2 g/mL
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- Amount(s) applied (volume or weight with unit): 10 ml/kg bw.

Duration of exposure:

24 h

Doses:

2000 mg/kg bw.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

yes

Remarks:

historical control.

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Systematic examinations were carried out daily to identify any behavioural or toxic effects on the major physiological functions during 14 days following the administration of the test item. The animals were weighed on day 0 (just before administering the test item) then on day 2, day 7, and day 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology (only if organs presenting abnormalities).

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No systemic clinical signs related to the administration of the test item were observed.

Gross pathology:

The macroscopic examinations of the animals at the end of the study did not reveal treatment-related changes.

Other findings:

- Other observations: Erythema was noted in all animals (10/10) at 24 hours post dose. This reaction was totally reversible at day 3. Dryness of the skin was noted in females at day 2 and in all animals at day 3. The skin recovered a normal aspect on day 4. A yellow coloration was noted in all animals at 24 hours post-dose.

Table 1. Body weight evolution

Males	D0	D2	D2-D0	D7	D7-D0	D14	D14-D0
Rm 0689	266	268	2	317	51	401	135
Rm 0690	271	273	2	317	46	381	110
Rm 0691	271	271	0	327	56	379	108
Rm 0692	258	265	7	315	57	384	126
Rm 0693	272	276	4	333	61	379	107
Mean	267.6	270.6	3.0	321.8	54.2	384.8	117.2
SD	5.9	4.3	2.6	7.8	5.8	9.3	12.6
Females
Rm 0694	217	213	-4	242	25	259	42
Rm 0695	229	224	-5	258	29	274	45
Rm 0696	226	222	-4	253	27	270	44
Rm 0697	222	221	-1	247	25	279	57
Rm 0698	216	213	-3	239	23	283	67
Mean	222.0	218.6	-3.4	247.8	25.8	273.0	51.0
SD	5.6	5.2	1.5	7.8	2.3	9.2	10.7

 

Table 2. Necropsy data sheet.

Observations	Males Rm0689 to Rm0693	Females Rf0694 to Rf0698
General appearance	Normal	Normal
Oesophagus	NTR	NTR
Stomach	NTR	NTR
Duodenum	NTR	NTR
Jejunum	NTR	NTR
Ileon	NTR	NTR
Caecum	NTR	NTR
Colon	NTR	NTR
Rectum	NTR	NTR
Spleen	NTR	NTR
Liver	NTR	NTR
Thymus	NTR	NTR
Trachea	NTR	NTR
Lungs	NTR	NTR
Heart	NTR	NTR
Kidneys	NTR	NTR
Urinary bladder	NTR	NTR
Ovaries	-	NTR
Uterus	-	NTR
Testicles	NTR	-
Treatment area (skin)	NTR	NTR
Adrenals	NTR	NTR
Pancreas	NTR	NTR
Particulars	None	none

Interpretation of results:

GHS criteria not met

Remarks:

EU criteria.

Conclusions:

The LD50 of the test item is higher than 2000 mg/kg body weight by dermal route in rats.

Executive summary:

The acute dermal toxicity of the test item was studied according to OECD 402 (GLP study). A limit test was performed by dermal administration of the test item at a dose of 2000 mg/kg bw to 5 male and 5 female Wistar (SPF:Caw) rats. After 24 h of exposure, the area was rinsed with water, and the animals were observed daily for 14 days. Observations included clinical signs, mortality, body weights, gross pathology and, if any abnormality was observed, histopathology of the affected tissues. No mortality or systemic clinical signs related to the administration of the test item were observed throughout the study. Erythema was noted in all animals (10/10) at 24 hours post dose. This reaction was totally reversible at day 3. Based on the results of the study, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study has a Klimisch score of 1.

Additional information

Oral route: Weight of evidence:

- Read-across from analogue substance. The acute oral toxicity of neohesperidin dihydrochalcone on rats was studied by a method similar to OECD TG 420. A limit test was performed by administering a single dose of 5g/kg bw test item to 10 (5M/5F) Wistar rats, and observing the effects for 14 days (clinical signs, body weights). No effects were observed in any animal. Therefore, the analogue substance was found to be non toxic, with an LD50 > 5000 mg/kg bw. Based on read-across approach, the oral LD50 in rats for the target substance was determined to be >= 4984 mg/kg bw.

- Read-across from analogue substance. The study of the acute oral toxicity of neohesperidin dihydrochalcone on rats was performed by a method similar to OECD TG 420. A limit test was performed by administering a single dose of 5g/kg bw test item to 10 (5M/5F) Wistar rats, and observing the effects for 14 days (clinical signs, body weights). One male died on day 9, but no gross changes were observed, and no effects were noted on any other animal. Therefore, the analogue substance was found to be non toxic, with an LD50 > 5000 mg/kg bw. Based on read-across approach, the oral LD50 in rats for the target substance was determined to be >= 4984 mg/kg bw.

- Read-across from analogue substance. During an in vivo micronucleus assay on the analogue substance neohesperidin dihydrochalcone, male and female Swiss-Webster mice were treated with the test substance over the range of 200-5000mg/kg, and observed for 48h. Under test conditions, no toxicity signs were observed on mice treated with the test substance. Therefore, the analogue substance is considered to be non-toxic, with an LD50 > 5000 mg/kg bw. Based on the read-across approach, the target substance is non toxic, with an LD50 > 4984 mg/kg bw.

Dermal route:

- Key study. The acute dermal toxicity of the test item was studied according to OECD 402 (GLP study). A limit test was performed by dermal administration of the test item at a dose of 2000 mg/kg bw to 5 male and 5 female Wistar (SPF:Caw) rats. After 24 h of exposure, the area was rinsed with water, and the animals were observed daily for 14 days. Observations included clinical signs, mortality, body weights, gross pathology and, if any abnormality was observed, histopathology of the affected tissues. No mortality or systemic clinical signs related to the administration of the test item were observed throughout the study. Erythema was noted in all animals (10/10) at 24 hours post dose. This reaction was totally reversible at day 3. Based on the results of the study, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Justification for classification or non-classification

Based on available data, the substance is not classified for acute toxicity according to CLP Regulation (EC) No. 1272/2008.