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  • Title: [Toxicological assessment on safety of water and 70% ethanolic extracts of nearly ripe fruit of Evodia rutaecarpa].
    Author: Yang XW.
    Journal: Zhongguo Zhong Yao Za Zhi; 2008 Jun; 33(11):1317-21. PubMed ID: 18831217.
    Abstract:
    OBJECTIVE: To study the acute toxicity and mutagenic risk of the water extracts (ERWE) and 70% ethanol extracts (EREE) from the nearly ripe fruit of Evodia rutaecarpa, and provide experimental basis for safety evaluation of ones. METHOD: The ERWE and EREE were prepared from the nearly ripe fruit of E. rutaecarpa by reflux extraction with H2O and 70% ethanol aqueous solution for three times, respectively. According to the terms from "technical standards for test & toxicological assessment of health food" issued by Healthy Ministry of PRC, acute toxicity, and Ames, mouse marrow cell micronucleus and mouse sperm aberration test were performed. Acute toxicity test of ERWE and EREE in mice was studied by the method of Horn to give the median lethal dose (LD50). Forty healthy Kunming strain male and female mice were used and their body weights ranged from 17-22 g. All of them were distributed randomly to 4 different dose groups which each had 10 mice. The ERWE or EREE was administered at the doses of 1.00, 2.15, 4.64 and 10.00 g x kg(-1), respectively, via intragastrical route. The number of animals poisoned and died in each group were noted daily for 7 consecutive days. The Ames test was carried out using the Salmonella typhimurium strain TA97, TA98, TA100 and TA102. In the sperm abnormalities test, 25 healthy adult male Kunming strain mice with a body weights ranged from 25-35 g were distributed randomly to 5 different groups (1 positive control, 1 negative control and 3 treated groups) which each had 5 mice. A single dose of 60 g x kg(-1) of cyclophosphamide was intragastrically administered to mice in a positive control group, and the mice in the negative control group were administered with the same volume of distilled water. In the treated groups, the ERWE or EREE was intragastrically administered at the doses of 1.25, 250 and 5.00 g x kg(-1), respectively, via the same route with the positive control group. The administration was carried out once daily for 5 consecutive days. The sperm suspension was prepared from caudal epididymis of male mice at 35th day after treatment with different doses of the extract. The suspension was stained with Eosin-Y and air-dried smears were prepared. One thousand sperms per animal were analysed for abnormal shapes and the rates of sperm aberration was calculated. In the mouse bone marrow micronucleus assay, 50 healthy adult male and female Kunming mice, weighing 25 to 30 g, were randomly assigned to five groups (1 positive control, 1 negative control and 3 treated groups) which each had 10 mice, five males and five females. The mice were intragastrically administered twice at intervals of 24 h with the ERWE or EREE at doses of 1.25, 2.50 and 5.00 g x kg(-1) in the positive control group. A single dose of 60 g x kg(-1) of cyclophosphamide in a positive control group and the same volume of distilled water in a negative control groups were intragastrically administered, respectively. Mouse bone marrow was obtained from 10 animals for each group at 6 h after the last dose administration. Smears were stained with Giemsa and analysed for the presence of mouse bone marrow micronucleus from 1 000 cells. RESULT: The oral acute toxicity study in mice revealed that the LD50 of the both ERWE and EREE was more than 10.0 g x kg(-1). The mice with both the poisoned sign or died had not been observed after intragastrical administration of ERWE or EREE at the doses of 1.00, 2.15, 4.64 and 10.00 g x kg(-1). The results of genotoxicity tests were all negative, including Ames, mouse marrow cell micronucleus and mouse sperm aberration test. In the all assay in vivo, the mice showed a normally progressive increase in body weight from the start to the end of the experiment. CONCLUSION: The oral LD50 of the ERWE and EREE in mice was more than 10.0 g x kg(-1) belonging to non-toxicity on the acute toxicity rating criteria. The both ERWE and EREE showed no genotoxicity in the experimental condition.
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