【作者】 张一贞；

【导师】 韩崇选； 张宏利；

【作者基本信息】 西北农林科技大学 ， 森林保护学， 2007， 硕士

【摘要】 本研究通过对不同生长期皂荚(Gleditsia sinensis Lam)的总皂苷含量及对小鼠的适口性,乙醇与丙酮提取液对小鼠的毒性,提取分离所得的总皂苷及精制皂苷对小鼠的毒性测定等,系统研究了皂荚活性物质的提取工艺及其对小鼠的毒杀活性,为把皂荚开发为一种无公害植物源灭鼠剂提供了理论依据与具体线索,研究结果如下:(1)测定了不同生育期的皂荚总皂苷含量通过分光光度法测得8、9、10月份皂荚中总皂苷含量分别为5.75%、22.19%、11.37%。(2)测定了皂荚的乙醇与丙酮提取液对小鼠的毒性大小采用灌胃法测定两中提取液对小鼠的毒性,结果证明,乙醇提取液对小鼠的0致死的最大量(Dn)和100%致死的最小量(Dm)分别为1.5 g·kg-1、4.0 g·kg-1,致死中量LD50=2.21 g·kg-1;丙酮提取液体的Dn=2.0 g·kg-1,Dm=6.0 g·kg-1,LD50=3.52 g·kg-1。通过对致死中量测定所得结果分析,乙醇提取液的毒力大于丙酮提取液的毒力,且两者毒力均较强。(3)测定了不同生育期皂荚干样对小鼠的毒性与适口性把8、9、10月分三个不同生育期的皂荚粉碎后拌入饵料,在饵料中皂荚含量均为15%,三个生育期皂荚对试鼠的毒杀率分别为70%、80%和90%;表明皂荚可导致试鼠中毒,且中毒死亡率较高。饵料中加入含量为15%的8、9、10月份三个不同生育期的皂荚时,小鼠对饵料的摄取量分别为0.27、0.29和0.48。由此可知,饵料中加入8、9月份皂荚皂荚样品后,对小鼠的适口性一般;饵料中加入10月份皂荚样品好,对小鼠有较好的适口性。(4)皂荚活性物质的提取分离实验通过70%乙醇提取,石油醚、乙酸乙酯、正丁醇萃取等方法得到总皂苷。把总皂苷进行硅胶柱层析分离,分别得到精制皂苷D1=2.07g,D2=0.62g,D3=10.28g。E=5.51g。对样品D3与E做高压液相色谱分析,结果显示,D3由12个化合物组成,其中最大含量的化合物为46.3%,E由15个化合物组成,最大含量的化合物为17.84%。(5)对总皂苷与精制皂苷进行杀鼠活性实验结果总皂苷对小鼠的100%致死最小剂量为2.0 g·kg-1,0致死最大剂量为0.8g·kg-1 ,致死中量LD50=1.3798 g·kg-1;其对试鼠的毒杀作用具有性别差异,对雄鼠的毒杀率大于雌鼠;在高剂量和低计量下表现出亚急性灭鼠剂特征,在中剂量下表现出急性灭鼠剂特征;且对雌鼠的急性毒性随着剂量的变化差异不是很大,而对雄鼠的急性毒性随着剂量变化产生极大差别。精制皂苷D3对小鼠的100%致死最小剂量为1.4g·kg-1,0致死最大剂量为0.6g·kg-1,致死中量LD50=0.8700 g·kg-1;其对试鼠的毒杀作用也具有性别差异,对雄鼠的毒杀率大于雌鼠;且给药剂量的变化对雌雄鼠的极性毒性都有所影响。雌鼠的剂量和平均致死时间的关系大致成正弦状,当剂量增大时,平均致死时间先变长,达到一定值后开始开始缩短;而对雄鼠的急性毒性也大致成正弦状,与雌鼠不同的是,雄鼠的平均致死时间变化具有两个拐点。且最长和最短平均致死时间的差值达到14倍。更多还原

【Abstract】 The extract technique of active material in Gleditsia sinensis Lam and poisonous function of the material on mice were studied to provide the theory to develop the Gleditsia sinensis Lam as a non-pollutant botanical rodenticide. In this study, the total quantity of saponin in different growth stage of Gleditsia sinensis Lam and palatability in mice were determined; determination of toxicity of the extracts by ethanol and acetone, toxicity of total and purifier saponin to mice were studied. Results are as below:(1)Determination of total saponin of Gleditsia sinensis Lam in different growth stage.The total saponin in Gleditsia sinensis Lam in different growth stage, August, September and October determined by colorimetric assay is 15.75%、22.19%、11.37%。(2)Determination of the toxicity of ethanol extract and acetone extract to mice in Gleditsia sinensis LamThe toxicity of both extracts was assayed by gastric lavage. The results showed that the Dn and Dm of ethanol extract were separately 1.5 g·kg-1and 4.0 g·kg-1, LD50=2.21 g·kg-1. While those of acetone extract were Dn=2.0 g·kg-1, Dm=6.0 g·kg-1, and LD50=3.52 g·kg-1. The toxicity of ethanol extract is stronger than the acetone extract after analyzing the results of LD50.(3)Determination of toxicity and palatability of dried samples of Gleditsia sinensis Lam to miceThe Gleditsia sinensis Lam in different growth stage, August, September and October, were mixed after pulverizing in bait. When the quantity of Gleditsia sinensis Lam in bait were all 15%, the killing rates of three kinds of Gleditsia sinensis Lam were separately 70%, 80% and 90%, which indicate Gleditsia sinensis Lam poisons mice and the killing rate is higher.At the same conditions, the bait quantities that mice ate were separately 0.27, 0.29and 0.48. Thus, to mix Gleditsia sinensis Lam sample from August and September in bait comes to a common palatability to mice, while mixing the sample from October, the palatability to mice was better.(4)Extraction and separation active material in Gleditsia sinensis Lam Total saponin was obtained by extraction with 70% ethanol, petroleun ether, ethyl acetate and n-Butanol, then was separated by silica-gel column chromatography to get purifier saponin (D1=2.07g, D2=0.62g, D3=10.28g; E=5.51g).(5)Rat-killing activity experiment with total saponin and purifier saponinThe Dn and Dm of total saponin were separately 2.0 g·kg-1 and 0.8g·kg-1, which LD50 was 1.3798 g·kg-1. The poisonous function depended on the sex of mice; the killing rate to male is greater than female. Subacute rodenticide is showed in high and low dose, while acute was found in moderate dose. The change of acute poison to female was not great with dose, while there was great difference to the change of acute poison to male with dose.The Dn and Dm of purifier saponin D3 is 1.4g·kg-1 and 0.6g·kg-1, which LD50 was 0.8700 g·kg-1. The poisonous function depended also on sex; the killing rate to male is greater than female. The change of dose effected acute poison to both sexes. The relationship between Dose and mean death time to female was like sine curve; the mean death time was increased, and then decreased with the dose increased. To female, the relationship was about the same except that there were two peaks with dose increased, and the Dm is 14-fold higher than the Dn. Thus, the effect of the acute poison of purifier saponin D3 to male was greater.更多还原