2,20:3,19-Dimethano-2,3,4a,5a,6a,7a,8a,9a,10a,11a, 12a,13a,14a,15a,16a,17a,19 ,20,21a,22a,23a,24a,25a,26a,27a,28a,29a,30a,31a,32a,33a,34a-dotriacontaazabispentaleno [1''',6''':5'',6'',7''] cycloocta[1'',2'',3'':3'',4'']pentaleno[1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-gh:1',2',3'-g'h']cycloocta[1,2,3-cd:5,6,7-c'd']dipentalene-1,4,6,8,10,12,14,16,18,21,23,25,27,29,31,33-hexadecone, hexadecahydro-, stereoisomer,2,18:3,17-Dimethano-2,3, ,4a,5a,6a,7a,8a,9a,10a,11a,12a,13a,14a,15a,17,18,19a, 20a,21a,22a,23a,24a, 25a,26a,27a,28a,29a,30a-octacosaazabispentaleno[1''',6''':5'',6'',7'']cycloocta [1'',2'',3'':3'',4''] pentaleno[1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-cd:1',2',3'-gh]pentalene-1,4,6,8,10,12,14,16,19,21,23,25,27,29-tetradecone, tetradecahydro-, stereoisomer, 1H,4H,12H,15H-2,14:3,13-Dimethano-5H,6H,7H,8H,9H,10H,11H,16H,17H,18H,19H,20H,21H,22H-2,3,4a,5a,6a,7a,8a,9a,10a,11a,13,14,15a,16a,17a,18a,19a,20a,21a,22a-eicosaazabispentaleno [1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-cd:1',2',3'-gh]pentalene-1,4,6,8,10,12,15,17,19,21-decone, decahydro-, stereoisomer

Endpoint summary

Description of key information

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

LC50 (rat, inhalation, 4h) > 5.35 mg/L

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Route of administration:

oral: gavage

Vehicle:

other: Methyl cellulose 1%

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

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

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:

LD0

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

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020/21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 433 (Acute Inhalation Toxicity: Fixed Concentration Procedure)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed concentration procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

>= 2.7 - <= 2.8 µm

Geometric standard deviation (GSD):

>= 2.33 - <= 2.58

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

targeted: 5 mg/l, achieved: 5.35 mg/l

No. of animals per sex per dose:

1 male and 1 female for the sighting study, 5 male animals for the main study

Control animals:

no

Preliminary study:

In 1 male and 1 female absence of mortality or toxicity at a target exposure level of 5 mg/L

Key result

Sex:

male

Dose descriptor:

LC0

Effect level:

5.35 mg/L air

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

salivation

Body weight:

On the day following exposure (Day 2), body weight loss was evident in 4/5 males exposed to 5.35 mg/L. The group mean body weight was comparable to Day 1 values by the next weighing occasion (Day 7) and continued to increase for the remainder of the observation period.
The changes in body weight gain on Day 2 are considered to be a consequence of the exposure (duration and removal of food and water), and therefore not test item-related.

Gross pathology:

The macroscopic examination after a single exposure followed by a fourteen day observation period for males exposed to 5.35 mg/L revealed no abnormalities.

Salivation was seen in 2/5 males on return to home cage, resolving by the end of the working day check.

Interpretation of results:

GHS criteria not met

Conclusions:

The LC50 (4-hour) is in excess of 5.35 mg/L for male rats.

Executive summary:

There were no test item-related deaths, macropathology findings or effects on body weight in an acute nose-only inhalation study in rats in accordance with OECD guideline 433. Under the conditions of this study the LC50 (4-hour) is in excess of 5.35 mg/L for male rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC0

Value:

5.35 mg/L air

Physical form:

inhalation: aerosol

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 and inhalation toxicity according to the CLP Regulation (EC) No 1272/2008.