Registration Dossier

Administrative data

Description of key information

Acute Oral Toxicity: LD50 >5000 mg/kg bw female Wistar rats (read-across from analogue substance).

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:
09 September 2014 to 24 September 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
EXPERIMENTAL ANIMALS
Species and strain: CRL: (WI) rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633, Sulzfeld
Hygienic level at arrival: SPF
Hygienic level during
the study: Standard housing conditions
Number of animals: 3 animals
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, ~8 weeks old
Date of receipt: 28 August 2014
Body weight at treatment: 210 – 223 g
Acclimation period: At least 12 days

Husbandry
Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 522/11
Housing: group caging†
Cage type: Type II polypropylene/polycarbonate
Bedding: Lignocel Bedding for Laboratory Animals was available to animals during the study.
A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal
rodent activities. The temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply
Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is
considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
The batch of feed employed in the study was as follows: 190 1786, expiry date: January 2015
The supplier provided an analytical certificate for the batch used. Copy of the certificate will be archived with the raw data. Water quality control
analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public
Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at
CiToxLAB Hungary Ltd.

Animal Identification
Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File, for each animal allocated to the treatment groups. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.
Route of administration:
oral: gavage
Vehicle:
other: Methyl cellulose 1%
Details on oral exposure:
A single oral gavage administration was followed by a fourteen-day observation period. On the day before treatment, the animals were fasted. The
food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral
gavage in the morning. The food was returned 3 hours after the treatment.
Doses:
The starting dose level (5000 mg/kg bw) was selected at the request of the Sponsor.
Initially, one female was treated at a dose level of 5000 mg/kg bw of CUCURBIT[8]URIL. The test item did not cause mortality; therefore further two
animals were treated at the same dose level. The test item did not cause mortality in these animals, so no further testing was required according to
OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.
No. of animals per sex per dose:
3 female rats were exposed to a single dose of 5000mg/kg bw
Control animals:
no
Details on study design:
Clinical Observations
Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual
observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system,
somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement
The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.

NECROPSY
Macroscopic examination was performed on all animals. The surviving animals were sacrificed by exsanguination under pentobarbital anaesthesia
(Euthasol® 40 %; details are presented in 3.1.3.). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

EVALUATION OF THE RESULTS
The method used was not intended to allow the calculation of a precise LD50 value.
The test item was ranked into categories of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423. Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.
Statistics:
None specified
Preliminary study:
Not applicable
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
CUCURBIT[8]URIL did not cause mortality at the dose level of 5000 mg/kg bw.
Clinical signs:
Treatment with CUCURBIT[8]URIL at a dose level 5000 mg/kg bw did not cause any test item related effects on the animals.
Body weight:
Body weight gains of CUCURBIT[8]URIL treated animals during the study showed no indication of a treatment-related effect.
Gross pathology:
There was no evidence of the macroscopic observations at a dose level of 5000 mg/kg bw.
Other findings:
None specified
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the test item CUCURBIT[8]URIL was found to be above 5000 mg/kg bw in female
CRL: (WI) rats. According to the GHS criteria, CUCURBIT[8]URIL can be ranked as “Unclassified” for acute oral exposure.
Executive summary:

The objective of the study was to assess the toxicity of test item CUCURBIT[8]URIL when administered as a single oral gavage dose to rats. The results of the study allow the test item to be ranked according to most classification systems currently used. Study performed in accordance with the study plan, OECD 423 (17th December 2001), Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris, EPA Health Effects Test Guidelines (OCSPP 8770.1100), United States, EPA 712-C-98-190 (1998) and the Principles of Good Laboratory Practice (Hungarian GLP Regulations: 42/2014. (VIII. 19.) EMMI decree of the Ministry of Human Capacities which corresponds to the OECD GLD, ENV/MC.CHEM (98)17).

Under the conditions of this study, the acute oral LD50 value of the test item CUCURBIT[8]URIL was found to be above 5000 mg/kg bw in female CRL: (WI) rats. According to the GHS criteria, CUCURBIT[8]URIL can be ranked as “Unclassified” for acute oral exposure.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
A GLP study was performed with an analogue substance according to OECD Guideline 423, EU Method B1. Tris and US EPA Procedure OCSPP 870.1100

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The single-dose oral toxicity of Cucurbit[8]uril hydrochloride hydrate was performed according to the acute toxic class method in CRL: (WI) rats. Initially, one female was treated at a dose level of 5000 mg/kg bw by gavage after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was administered formulated in Methyl cellulose 1% at a concentration of 500 mg/mL at a dosing volume of 10 mL/kg bw. The test item did not cause mortality and two further animals were treated at the same dose level. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and Day 14 before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

The test substance did not cause mortality in these animals, and did not cause any test substance related effects on the animals. There were no changes in body weight gains or any macroscopic findings. Therefore, no further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris. Under the conditions of this study, the acute oral LD50 value of Cucurbit[8]uril hydrochloride hydrate was found to be above 5000 mg/kg bw in female CRL: (WI) rats.


Justification for classification or non-classification

Based on the available data, the test substance does not require classification for acute oral toxicity according to the CLP Regulation (EC) No 1272/2008.