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Administrative data

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:
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 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:
No effects observed.
Body weight:
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:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Remarks:
(SPF Caw)
Sex:
male/female
Details on test animals 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:
No systemic clinical signs related to the administration of the test item were observed.
Body weight:
The body weight evolution of the animals remained normal throughout the study.
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.