safety

Oral toxicity experiment using mouse

We have had Kippogen searched at the Japanese Health Food Center based on the OECD Guidelines for Testing of Chemicals 401(1981)

◆ Adjusting the test liquid
By using a muller, we have grinded the sample and used the suspended substance which went through the opening of 150um. By using distilled water, we have adjusted the test liquid to 200mg/ml.
◆ Tested animals
We have bought both ICR male and female mouse of the age of 4 weeks. After breeding them for a week and checking that they are in normal condition, we had them tested. With the room temperature of 23℃±2℃, 12 hours of illumination per day, 5 mice were bred. Food and water was given any time they wanted.
◆ Method of Experiment
We have tested 10 mice both male and female for tested group and controlled group. Before testing we had them fasted for 4 hours. After measuring their weight, the tested group that had been administered 5,000mg/kg of dosage as specimen administration amount using gastroptosis sonde with oral toxity. As for the controlled group, distilled water was given, 0.8ml for male, 0.7ml for female each. The observation period was 2 weeks, making observation once a day, administering as many times per possible. We had them weighed on the 7th and 14th day and compared both groups by 5% significance level of t-test. We had all the animals autopsied after the observation period.
◆ Test Results
    1) Death report
    No death was reported during the test period.
    2) Daily report
    No abnormal situation was reported during the test period.
    3) Weight change(Table 1 – and2)
    After administration there was no change of weight increase between both male and female groups.
    4) Autopsy report
    There was no abnormal situation reported in the organs of both male and female after autopsy.
◆ Study
Based on the guidelines of OECD, Guidelines for Testing of Chemicals 401 (1981), we have enforced an acute toxicity experiment to the mouse. According to the guideline, if a death case is seen at 5,000mg/kg dosage, a detailed test to exam the amount of LD50 is instructed.
However, in this experiment death case was not reported, neither was there any abnormal situation reported after autopsy. Therefore, it can be considered that the LD 50 amount in single oral administration both male and female was over 5,000mg/kg.