oligomeric Reaction products from Hydrogenation and Hydrolysation of Starch

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Dec 2004 - Sept 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

473-690-8

EC Name:

-

Cas Number:

738602-93-2

Molecular formula:

not applicable for UVCB

IUPAC Name:

(2R,3R,4S,5S,6R)-2-{[(2R,3R,4R,5S,6R)-5-{[(2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}oxan-2-yl]oxy}-3,4-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-6-(hydroxymethyl)oxane-3,4,5-triol; (2S,3R,5R)-4-{[(2R,3R,4R,5S,6R)-5-{[(2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}oxan-2-yl]oxy}-3,4-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}hexane-1,2,3,5,6-pentol

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: 4G08
- Purity, including information on contaminants, isomers, etc.: 54.2 % Main component, 9.4 % other glucosides 36.4 % other Polyols

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature away from direct sunlight, seal tightly and store in exsiccator
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: stable under storage conditions
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: assumed stable
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: assumed stable
- Reactivity of the test material with the incubation material used (e.g. plastic ware): assumed stable

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

recognized by the international guidelines as the recommended test system

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC LtD, CH-4414 Füllinsdorf, Switzerland
- Females (if applicable) nulliparous and non-pregnant: not stated
- Age at study initiation: 6 weeks
- Weight at study initiation: Males 138.7 - 159.3 g
Females 115.8 - 129.2 g
- Fasting period before study: no
- Housing: in groups of 5
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

DETAILS OF FOOD AND WATER QUALITY:
Community tap-water was used
standard rat maintenance diet was used

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +- 3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

Administration by gavage is a common and accepted route of exposure for studies of this type.

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The dose formulations were prepared daily. MG-60 was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume). the mixtures were prepared using a magnetic stirrer and stored at room temperature

VEHICLE
- Justification for use and choice of vehicle (if other than water): not relevant
- Concentration in vehicle: not stated
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): not required
- Purity: not relevant

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration, homogenity and stability (after 2 hours) of the dose formulations were determined in samples taken after experimental start. Concentration and homogeneity of the dose formulations were determined in samples taken during week 3 of the treatment. The analyses were perfromed by RCC Ltd according to a HPLC method supplied by the sponsor.

Duration of treatment / exposure:

28 d

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

50 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

200 mg/kg bw/day (actual dose received)

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based upon the results of a non-GLP 5-day dose range finding study in which MG-60 was administered by gavage to 2 rats per group and sex. The treatment was well tolerated and did not produce clinical signs or early mortality. There were no relevant treatment-related effects on food intake, body weight development, macroscopic appearance and organ weights.
- Rationale for animal assignment (if not random): random
- Fasting period before blood sampling for clinical biochemistry: 18 hours
- Rationale for selecting satellite groups: no satellite group
- Post-exposure recovery period in satellite groups: not relevant

Positive control:

none

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality, daily for clinical signs

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
not relevant

OPHTHALMOSCOPIC EXAMINATION: No
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 4 weeks
- Anaesthetic used for blood collection: Yes (identity) isoflurane
- Animals fasted: Yes
- How many animals: all animals


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 4 weeks
- Animals fasted: Yes
- How many animals: all animals

PLASMA/SERUM HORMONES/LIPIDS: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: during week 4
- Dose groups that were examined: all
- Battery of functions tested: grip strength / locomotor activity

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Optional endpoint(s):

Optional endpoints: No

Statistics:

The following statistical methods were used to analyze the grip strength, locomotor activity, body weight, organ weights and ratios, as well as:
the Dunnett-test based on a pooled variance estimate were applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
Fisher's exact-test was applied to the macroscopic findings.
The following statistical methods were used for statistical analysis of clinical laboratory data:
Quantitative data were analyzed b a one-way analysis of variance (ANOVA) when the variances are considered homogeneous according to Bartlett. Alternatively, if the variances are considered to be heterogenous (p<= 0.05), a non-parametric Krukal-Wallis test was used. Treated groups were compared to the control groups using Dunnett's test if the ANOVA was significant at the 5 % level and by Dunn's test in the case of a significant Kruskal-Wallis test (p<= 0.05).

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

one control male presented soft feces during week 3. All the other animals presented no clinical signs dring weeks 1-3

Mortality:

no mortality observed

Description (incidence):

all animals survived until their scheduled necropsy

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No test item-related changes of toxicological relevance were noted in body weight and body weight gain of males and females

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

The mean food consumption and the relative food consumption of the test item-treated males and females compared favorably with their respective controls during the treatment period

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No changes of toxicological relevance were noted in the hematology parameters after the treatment period with the test item both in males and females.

Significant increases in neutrophiles concentrations were measured in males treated with 50 mg/kg/day (p<0.01) and 1000 mg/kg/day (p<0.01). Significant increases in the mean corpuscular Volume (MCV) and mean corpuscular hemoglobin (MCH) were noted in females treated with 200 mg/kg/day (p<0.05 and p<0.01, respectively). Although MCH Values were slightly above the range of the historical control data, this findling was neither dose dependent nur supported by similar changes in related parameters and, therefore,
considered to be an incidental Change without toxicological relevance. Neutrophiles and MCV Values remained within the range of the historical control data and were therefore considered to be incidental.
Both control and test item-treated females presented low levels of high fluorescence reticulocyte maturity Index (H-reti), which were all below the range of the historical control data. Females treated with 1000 mg/kg/day had significantly decreased H-Reti (p<0.05) but due to extreme control Values, this findling was not considered to be test item related.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

No changes of toxicological relevance were noted in the Clinical biochemistry parameters
after the treatment period with the test item both in males and females.
A dose dependent decrease in the total bilirubin (Bili-tot) concentrations was measured in
test item-treated males (p<0.05 at 50 mg/kg/day, p<0.05 at 200 mg/kg/day and p<0.01 at
1000 mg/kg/day) but, as the Values remained weithin the range of the historical control data
and were only observed in the individual gender, they were considered to be test item related
but of no toxicological relevance.
A significant increase in the triglycerides concentration was noted in males treated with 50
mg/kg/day but as the Value remained weithin the range of the historical control data and was
not observed in the high dose group, this finding was considered to be incidental.

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

No clinical signs of toxicological relevance were observed during week 4
Locomotor Activity
Significant increases in the total and Single runs locomotor activity (0-10 and 10-20 minutes time intervals) were measured in test item-treated males when compared to Controls.
Although these findlings were test item related, hey were not considered to be toxicologically relevant.
The locomotor activity of the test item-treated female rats was not affected.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Significant decreases in spleen-to-body weight (p<0.05) and spleen-to-brain weight (p<0.05) ratios were noted in males treated with 200 mg/kg/day after four weeks treatment.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

All findings recorded were considered to be within the range of normal background lesions that may be seen in ats of this strain adn age. They consisted of discoloured foci in the thymus (one control male) and pelvic dilation in kidneys (one male treated with 50 mg/kg/day).
Although these findlings were test item related, hey were not considered to be toxicologically relevant.
The females showed no macroscopic findings.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

All findings were within the range of sponaneous background lesions, which may be recorded in rats of this strain and age.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

No Histopathological findings: neoplastic reported

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

clinical biochemistry

clinical signs

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Target system / organ toxicity

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Based on the resultat of this Study, the no-observed-adverse-effect-level (NOAEL) could be
established at 1000 mg/kg body weight/day of MG-60.

Executive summary:

In this subacute toxicity Study, MG-60 was administered daily by oral gavage to SPF-bred
Wistar rats of both Sexes at dose levels of 50, 200 and 1000 mg/kg body weight/day for a
period of 28 days. A control group was treated similarly with the vehicle, bidistilled Vater,
only.
The groups comprised 5 animals per Sex, which were sacrificed after 28 days of treatment.
Clinical signs, outside cage Observation, food consumption and body weights were recorded
periodically during acclimatization and the treatment periods. Functional obsewational
batterie, locomotor activity and grip strength were performed during weck 4.
At the end of the dosing period, blood samples were withdrawn for hematology and plasma
Chemistry analyses. All animals were killed, necropsied and examined post mortem.
Histological examinations were performed on Organs and tíssues for all control and high
dose animals, and all gross lesions for all animals.

 

Oral administration of MG-60 to Wistar rats at doses of 50, 200 and 1000 mg/kg/day, for
28 days resulted in no mortalities, no Clinical signs of toxicological relevance during daily er
weekly observations (weeks 1-3) er during functional obsewational batterie (weck 4), no
effects on grip strength, food consumption er body weights and no changes of toxicological
relevance both in hematology and Clinical biochemistry parameters.
Test item-related findlings were generally restricted to increased total locomotor activity (in
mid- and high-dose groups) and Single runs locomotor activity (during the first 20 minutes of
measurement) in test item-treated males (at any dose level) and in decreased Spleen-to body
weight and Spleen-to brain weight ratlos in males treated with 200 mg/kg/day.
Based on the resultat of this Study, the no-observed-adverse-effect-level (NOAEL) could be
established at 1000 mg/kg body weight/day of MG-60.