Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication report which meets basic scientific principles.

Data source

Reference

Reference Type:

publication

Title:

Acute Oral Toxicity of Sucrose

Author:

Boyd, E.M. et al.

Year:

1965

Bibliographic source:

Toxicology and Applied Pharmacology 7:606-618

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Wistar rats were exposed to high dosages of sucrose ranging from 5000 - 80 000 mg/kg bw in a single application via gavage to calculate the LD50 value. Clinical signs and pathological findings were correlated to the time point of death of nonsurviving animals after gavage. The reversibility of treatment-related effects was established in surviving animals 2 and 4 weeks after substance administration.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sucrose

EC Number:

200-334-9

EC Name:

Sucrose

Cas Number:

57-50-1

Molecular formula:

C12H22O11

IUPAC Name:

beta-D-fructofuranosyl alpha-D-glucopyranoside

Details on test material:

- Name of test material (as cited in study report): sucrose
- Analytical purity: no data

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Department of Pharmacology, Queen´s University, Kingston, Ontario, Canada
- Age at study initiation: adult
- Weight at study initiation: 300-500 g (males), 250 - 300 g (females)
- Fasting period before study: Animals were fasted 16 h prior to administration (only food starvation, water was provided ad libitum).
- Housing: individual in metabolism cages
- Diet: Purina Fox Chow Checkers, Ralston Purina Company, St. Louis, Missouri, ad libitum
- Water: ad libitum

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: distilled water

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 60 mL/kg bw

MAXIMUM DOSE VOLUME APPLIED: 60 mL/kg bw

Doses:

5000 - 80 000 mg/kg bw (divided in 19 dosages), not further specified

No. of animals per sex per dose:

males: 48 (controls), 16 - 20 (mid-dose groups), 4 - 8 (the lowest 4 and highest 3 dose groups)
females: 30 (not further specified)

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days and 1 month
- Frequency of observations and weighing: animals were observed 6 days a week for 2 weeks followed by casual observations up to 1 month
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, gross and histopathology, other: food and water intake, colonic temperature, determination of water levels in the organs, urinalyses.
To determine the above mentioned parameter, the following organs were examined: adrenal glands, brain, gastrontestinal tract including cardiac and pyloric stomach, small bowel, caecum and the colon, heart, kidneys, liver, lungs, muscle of the abdominal wall, skin, spleen, testes and the thymus gland in addition to the residual caracsses.

Statistics:

Statistical methods used were those described by Croxton (1953, Elementary Statistics with Applications in Medicine. Prentice-Hall, New York).

Results and discussion

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50

Effect level:

29 700 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

male

Dose descriptor:

LD50

Effect level:

35 400 mg/kg bw

Based on:

test mat.

Mortality:

In the lower dose groups ranging from 5 000 - 24 000 mg/kg bw, no deaths occured whereas all animals died from the high-dose groups (60 000, 70 000 and 80 000 mg/kg bw). The frequency distribution of deaths versus time of animals exposed to up to 50 000 mg/kg bw showed a peak at 4 h, a valley at 9 h and a broad plateau at 12 - 48 h. In detail, 76 deaths occured at 9 h or less, all except 1 animal given 35 000 mg/kg bw and more. 43 deaths occured beyond 9 h with 23 of them in animals given less than 35 000 mg/kg bw. 38 deaths occured within 10 - 48 h and 5 deaths within 2 - 11 days. In addition, 2 control animals died on the 5th day possibly due to the toxic effects of distilled water.

Clinical signs:

In general, dehydration of test animals was observed following administration. Further, initial clinical signs including hypokinesia, prostration, abdominal bloating and diarrhea were observed in test and control animals with controls showing the symptoms to a lesser extent and period of time. In detail, test animals showed stronger effects during the first 6 h which persisted to 24 - 48 h. At that time, controls were already free of clinical signs. In addition, cyanosis was observed only in test animals. Further, gastroenteritis appeared after 1 - 3 h and disappeared at 1 - 4 days.
Premortal signs were tonic-clonic convulsions followed by stupor and respiratory failure observed in animals that died within 9 h after gavage.

Body weight:

Animals exposed to more than 20 000 mg sucrose/kg bw revealed a significantly decreased body weight ( p < 0.001) which was due to anorexia and diarrhoe. Moreover, food intake was depressed during the first 24 h following administration in all surviving rats given dosages above 20 000 mg/kg bw whereas the water intake rose in rats given dosages up to 20 000 mg/kg bw and varied thereafter erratically with a second peak in animals exposed to approx. 45 000 mg/kg bw.

Gross pathology:

In parallel to the generalized dehydration and loss of body weight, the following findings were detected in nonsurviving animals: severe but temporary gastroenteritis, a temporary mild hepatitis, marked nephritis and encephalities, focal necrosis of the heart, moderate stressor reactions in the adrenal and thymus gland, inhibition of spermatogenesis and arteriolitis. In contrast, the parenchyma of the lungs, pancreas, salivary glands, spleen and the skin appeared normal.
Organs of surviving animals appeared grossly normal.

Other findings:

Organ weights: the autopsy of 16 nonsurviving test animals (dose groups are not further specified) showed a significant loss of wet weight in the carcasses (- 13.9%), the liver (- 42.6%) and testes (- 7.2%). Moreover, water levels decreased significantly in the carcass (- 28.9%) and the following organs: pyloric stomach (- 39.6%), small bawel (- 25.7%), caecum (- 43.5%), colon (- 27.2%), lungs (- 22.5%), muscle of the abdominal wall (- 19.6%), skin (- 24.2%) and testes (- 15.8%). In contrast, a significant increase in wet weight was observed for the small bowel of the gastrointestinal tract (+ 16.4%). No significant increases in the water levels were observed.
In surviving test animals, significantly increased organ weights were determined 2 weeks after substance administration for the adrenal gland (+ 35.5 %), stomach (cardiac stomach: + 29.4%, pyloric stomach: + 25.9%), small bowel (+ 52.7%), caecum (+ 66.3%) and striated muscle (+ 27.8%) while the weights of the heart (- 14.6%), kidney (- 12.8%), liver (- 14.4%) and spleen (-20.5%) were significantly decreased. 1 months after substance administration, a significant decrease in liver wet weight was determined (- 26.1%). Significant low water levels were found in the brain (- 3.7%), the polyoric stomach (- 7.2%) and the spleen (- 10.8%).

Histopathology of nonsurviving animals revealed the following findings (the given time reflects the time frame between substance administration and death):
gastrointestinal tract: Gastroenteritis appeared at 1 - 3 h and disappeared at 1 - 4 days and was confined mostly to the stomach, the small bowel and the cardiac stomach with capillary, arteriolar and venous congestion of the lamina propria and submucosa. Areas of lysis of the surface layers of stratified squamous epithelium was observed in the cardiac stomach. Along with a similar vascular reaction in the pyloric stomach, a marked lysis of the surface of columnar epithelium and moucous neck cells, somewhat less lysis of the parietal cells was observed. Further, shrinkage of the zymogens with swelling of the lamina propria at the base of the gastric glands was detected. The villi of the small bowel were congested, the lamin propria edematous and the columnar epithelium completely denuded in many areas with very little heamorrhage in the lumen. An inflammatory reaction in the caecum was confined to capillary congestion and edema of the lamina propria and in the colon to a mild capillary congestion. Animals that died at 24 h did not show alterations in the latter 2 structures while the upper part of the alimentary canal was largely normal. Animals that died 4 days after administration showed occasional mild congestion in the submucosa and some edema of the arteriolar walls.

liver: The marked loss of weight was correlated to a shrinkage of hepatic cells: the sinusoids were distended and filled with red blood cells. The Kupffer cells were shrunken and often lysed away from the lining of the sinusoids. This effect was mostly evident at 3 h. By 24 h, the liver appeared microscopically normal.

kidney:Already 1 h after administration of a lethal dose of sucrose, alterations in the kidney were observed including shrunken glomeruli, increased subcapsular space, shrunken and distorted proximal tubular cells, prominent granules at the base of the brush border. At 3 h, the lumen of the proximal convoluted tubular cells were swollen and obliterated. Further, the loop cells were edematous and the distal tubular cells were shrunken with casts in the lumen. Vacuolar degeneration of the proximal tubular cells appeared at 24 h and bits of tubular debris was detected refluxed into the glomerular space beneath Bowman´s capsule. In addition, at this time, the squamous epithelium lining in the descending limb of the loop of Henle was swollen and some necrosis of the cuboidal epithelium lining the ascending limb was evident. An inflammatory reaction was less marked in the distal convoluted and collecting tubular cells were shrunken with casts in the lumen. A vacuolar degeneration was observed in animals that died at 4 days.

coronary arteries: The coronary arteries were dilated and the arterial wall was swollen and pale stained 3 h after administration in nonsurviving animals. In parallel, a marked capillary vasodilatation of the myocardium with minute areas of capillary heamorrhage was present. 1 - 4 days after substance administration, minute areas of focal necrosis with leucocyte infiltration were found scattered throughout the heart muscle.

adrenal glands: 1 - 3 h after gavage, cortical sinusoids of the adrenal glands were either congested or collapsed. Subsequently, they appeared normal but enlarged.

thymus gland: The microscopic appearance of the thymus gland varied. However, common findings were mild capillary and venous congestion in parallel to some loss of thymocytes.

testes: 1 h after gavage, a shrinkage of the seminiferous tubules was evident. When death was delayed to 4 days, spermatogenic cells were found separated from the tubular wall. The sperm was shriveled and wrinkled. Further, leucocyte infiltration in the interstitial tissue was observed.

arterioles: In the lung, a periarteriolar edema was observed 1 h after the lethal dose was applied. At 3 h, the intima and media of the splenic arterioles were edematous and pale stained. In the heart and the mesentery, the edema extended to the tunica externa. In the blood stream, large numbers of leucocytes were recorded.

minor changes in nonsurvivors: Phagocytosis was active in macrophages in the thymus and the spleen. The M disks of striated muscle were prominent in animals dying at 1 h. Later the muscle fibers were swollen in some animals and shrunken in the others. Further, the subcutaneous tissues were shrunken. Microscopical analyses of animals that died 4 days after gavage indicated a reactive hypertrophy of the gastrointestinal mucosa which appearently counted for the increase in weight of the alimentary organs at day 14.

-Microscopical analyses of surviving animals: 1 months after substance administration, livers of surviving test animals were significantly smaller (wet weight: - 26.1% of controls). Further, the water content of the thymus gland showed a significant decrease around 28% compared to controls.


- Other observations:
Colonic temperature over the first 24 h after substance administration: An increased colonic temperature was determined in test animals exposed to up to 30 000 mg/kg bw sucrose. Test animals of higher dose groups showed erratically decreased colonic temperatures with the mean over all dosages above 30 000 mg/kg bw beeing significantly reduced (p = 0.005).

Urinalyses: A dose-dependent increase in the excretion of glucose and the volume of urine was observed in animals with a peak near the LD50 value (urine volume at approx. 30 000 mg/kg bw; glucose content at approx. 25 000 - 35 000 mg/kg bw). In test animals exposed to higher dosages, no dose-dependent trend was obvious in these parameters. Moreover, the daily urinary output of protein revealed no treatment-related effect. In contrast, the urinary pH was decreased in test animals in a dose-dependent manner.

Any other information on results incl. tables

REVERSIBILITY OF EFFECTS

The analyses indicated that the above mentioned clinical signs observed in nonsurviving animals were reversible in animals which survived the study. Clinical signs like hypokinesia, prostration, cyanosis, abdominal bloating, diarrhea, anorexia, glycosuria and weight loss disappeared by the 3rd day while polydispia and polyuria, the elevated body temperature and alkalinuria disappeared by the 6th day.

Potential target organs: brain and meninges, gastrointestinal tract, liver (seems to be capable to recover rapidly within 24 h in regard to histopathological findings)

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified
DSD: not classified