Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was performed between 25 September 2012 and 06 November 2012.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.
Test type:
up-and-down procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Sponsor's identification: Sodium Glucoheptonate
Description: dark brown liquid
Lot number: 921000100
Purity: 49.5%
Date received: 09 February 2012
Expiry date: 13 January 2013
Storage conditions: room temperature in the dark

The integrity of supplied data relating to the identity, purity and stability of the test item is the responsibility of the Sponsor.





Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals 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 animals 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 and identified by a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The bodyweights fell within an interval of ±20% of the mean weight of any previously dosed animals.

The animals were individually housed 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 U.K. 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°C, 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.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Remarks:
no vehicle was used at the top two concentrations however for the purpose of the 354 mg/kg dose level (equivalent to 175 mg active ingredient), the test item was freshly prepared, as required, as a solution at the appropriate concentration in water.
Details on oral exposure:
For the purpose of the 1112 and 4040 mg/kg dose levels (equivalent to 550 and 2000 mg active ingredient/kg bodyweight, respectively), the test item was used as supplied. The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level. For the purpose of the 354 mg/kg dose level (equivalent to 175 mg active ingredient/kg bodyweight), the test item was freshly prepared, as required, as a solution at the appropriate concentration in distilled water.

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 the fasted bodyweight at the time of dosing. Treatment of animals was sequential.

Sufficient time (at least 48 hours) was allowed between each individual animal to confirm the outcome of the previously dosed animals.
Doses:
354 mg/kg equivalent to 175 mg/kg of active ingredient
1112 mg/kg equivalent to 550 mg/kg of active ingredient
4040 mg/kg equivalent to 2000 mg/kg of active ingredient
No. of animals per sex per dose:
Initially 1 female was tested at a dose level of 354 mg/kg
A second animal was tested at a dose level of 1112 mg/kg
Then 3 females were tested at a dose level of 4040 mg/kg
Control animals:
no
Details on study design:
No information was available regarding the toxicity of the test item therefore the default values for LD50 and sigma were entered into AOT425 Statistical Program. The statistical program gave a recommended dose progression of 2000, 550, 175, 55.0, 17.5, 5.5 and 1.75 mg/kg.

The first animal was dosed at 354 mg/kg (equivalent to 175 mg active ingredient/kg bodyweight). Further animals were then treated as follows based on the short-term results of the previously treated animal:

The test was complete after the fifth animal had been dosed as the following stopping criterion was met:
three consecutive animals survived at the maximum dose level (4040 mg/kg, equivalent to 2000 mg active ingredient/kg bodyweight)

- Duration of observation period following administration: 14 days (or other?)
- Frequency of observations and weighing: The animals were observed for deaths or overt signs of toxicity 0.5, 1, 2 and 4 hours after dosing and subsequently, once daily for fourteen days. Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment.
- Necropsy of survivors performed: At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
- Other examinations performed: clinical signs, body weight,

Statistics:
No information was available regarding the toxicity of the test item therefore the default values for LD50 and sigma were entered into AOT425 Statistical Program. The statistical program gave a recommended dose progression of 2000, 550, 175, 55.0, 17.5, 5.5 and 1.75 mg/kg.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 4 040 mg/kg bw
Based on:
test mat.
Remarks on result:
other: (equivalent to 2000 mg active ingredient/kg bodyweight).
Mortality:
There were no deaths.
Clinical signs:
Hunched posture was noted in the animal treated at a dose level of 1112 mg/kg (equivalent to 550 mg active ingredient/kg bodyweight). No other signs of systemic toxicity were noted.
Body weight:
Animals showed expected gains in bodyweight over the study period, except for one animal treated at a dose level of 4040 mg/kg (equivalent to 2000 mg active ingredient/kg bodyweight) which showed expected gain in bodyweight during the first week but no gain in bodyweight during the second week.
Gross pathology:
No abnormalities were noted at necropsy.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be greater than 4040 mg/kg bodyweight (equivalent to 2000 mg active ingredient/kg bodyweight).
Executive summary:

Introduction:

The study was performed to assess the acute oral toxicity of the test item following a single oral administration in the Wistar strain rat. The method was designed to be compatible with the following:

• OECD Guidelines for the Testing of Chemicals No. 425 "Acute Oral Toxicity Up- and-Down-Procedure (UDP)" (adopted 03 October 2008)

Method:

A total of five female animals were dosed individually in sequence with sufficient time (at least 48 hours) between each animal, at dose levels of 354, 1112 or 4040 mg/kg bodyweight (equivalent to 175, 550 and 2000 mg active ingredient/kg bodyweight, respectively).

The test item was administered orally undiluted at dose levels of 1112 and 4040 mg/kg bodyweight (equivalent to 550 and 2000 mg active ingredient/kg bodyweight, respectively) and as a solution in distilled water at a dose level of 354 mg/kg bodyweight (equivalent to 175 mg active ingredient/kg bodyweight). Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality: There were no deaths.

Clinical Observations: Hunched posture was noted in the animal treated at a dose level of 1112 mg/kg (equivalent to 550 mg active ingredient/kg bodyweight). No other signs of systemic toxicity were noted.

Bodyweight:

Animals showed expected gains in bodyweight over the study period, except for one animal treated at a dose level of 4040 mg/kg (equivalent to 2000 mg active ingredient/kg bodyweight) which showed expected gain in bodyweight during the first week but no gain in bodyweight during the second week.

Necropsy: No abnormalities were noted at necropsy.

Conclusion:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be greater than 4040 mg/kg bodyweight (equivalent to 2000 mg active ingredient/kg bodyweight).