【作者】 唐国文；

【导师】 杨长举；

【作者基本信息】 华中农业大学 ， 动物学， 2007， 博士

【摘要】 本研究以豆科植物TFG为材料，以主要的储粮害虫谷蠹、玉米象、杂拟谷盗、锯谷盗为试验对象，对粗提物和分离物的触杀、驱避活性进行了跟踪测定，系统研究了溶剂和提取方法对粗提物触杀、驱避活性的影响，比较了提取率和生物活性的差异，客观评价了与提取效率及杀虫活性相关的因子在提取分离过程中的作用，并确立了一种用于提取TFG生物活性物质的最优方案，分离出其中所含的活性成分，鉴定出其化学结构，最后进一步研究了活性单体的作用。主要研究结果如下：1．以无水乙醇、氯仿、乙酸乙酯、丙酮、石油醚为溶剂采用温浸回流法对TFG种子中的生物活性物质进行提取。用乙醇作溶剂时，对TFG的提取率最高，为11.48％，TFG的石油醚提取率最低，只有4.91％。TFG的氯仿提取率、乙酸乙酯提取率和丙酮提取率分别为7.24％、7.54％和5.29％。生物测定结果表明：TFG的5种溶剂粗提物对玉米象成虫都表现出一定的驱避作用。氯仿温浸粗提物对玉米象的驱避效果最好，在剂量为100μg／cm²的驱避级别达到Ⅱ级，在处理剂量为5000μg／cm²时，驱避级别达到Ⅲ级，其次是丙酮粗提物，在处理剂量为100μg／cm²时对玉米象的驱避级别达到Ⅱ级。TFG的5种溶剂粗提物对杂拟谷盗的驱避作用均优于对玉米象的作用。在各粗提物剂量为1.6μg／cm²时，7天平均驱避率均超过80％，驱避等级为Ⅴ级，丙酮粗提物对杂拟谷盗的驱避效果最好，其次是氯仿粗提物，乙酸乙酯粗提物对杂拟谷盗的驱避效果最差。多重比较结果显示：TFG各粗提物对玉米象与锯谷盗的驱避率之间差异不显著；各粗提物对杂拟谷盗与玉米象的驱避率之间差异显著：各粗提物对锯谷盗与谷蠢的驱避率之间差异显著；TFG粗提物对4种仓虫成虫的驱避活性大小顺序为：杂拟谷盗＞玉米象＞锯谷盗＞谷蠹。TFG的5种溶剂粗提物对玉米象成虫都有较好的触杀作用，玉米象成虫在接触药膜3天后，转而用干净无虫的小麦粒饲养1周，几乎均已死亡。TFG乙醇粗提物的上层物质对玉米象的触杀活性最强，处理玉米象24h和48h后的致死中量(简称LD₅₀分别为0.362mg／cm²和0.328mg／cm²；乙酸乙酯粗提物和丙酮粗提物对玉米象的触杀活性也较强，丙酮粗提物处理玉米象24h后的LD₅₀为0.380mg／cm²，乙酸乙酯处理玉米象48h后的LD₅₀为0.334mg／cm²；触杀活性较低的是石油醚粗提物，处理玉米象24h和48h后得到的LD₅₀分别比乙醇粗提物的LD₅₀高2倍多。TFG的5种溶剂粗提物对杂拟谷盗成虫的触杀作用较弱，杂拟谷盗成虫在接触药膜3天后，转入试管内饲养1周，死虫的数量仅有小幅度上升。TFG乙醇粗提物上层物质的触杀效果最好，处理杂拟谷盗成虫24h、48h、72h和10d后的LD₅₀分别为0.482mg／cm²、0.450mg／cm²、0.400mg／cm²和0.386mg／cm²，对杂拟谷盗触杀活性最低的是石油醚粗提物。TFG的5种溶剂粗提物处理谷蠹24h后，丙酮粗提物触杀活性表现为最强，它对谷蠹成虫的LD₅₀为0.240mg／cm²，触杀效果略高于乙醇粗提物和氯仿粗提物；经各粗提物处理后，谷蠹的死亡率从24h到48h的上升幅度较大。从TFG的5种溶剂粗提物对锯谷盗触杀试验结果可知，随着时间的延长，锯谷盗死亡率的变化幅度因粗提物以及施用的药剂剂量的不同而不同，TFG乙醇粗提物上层物质和氯仿粗提物的触杀效果较好，触杀活性最低的是石油醚粗提物。2．以TFG种子干粉为原料，以无水乙醇为提取溶剂，采用冷浸法、温浸法、索氏法、冷浸-索氏法、超临界CO₂萃取法（SCFE）对TFG粗提物的提取方法进行了比较研究，SCFE法的提取率为11.35％，高于冷浸法（4.65％）和冷浸-索氏法（8.14％），略低于索氏法（12.99％），和温浸法（11.48％）的提取率基本持平。5种方法得到的粗提物对杂拟谷盗都表现出良好的驱避性，不同粗提物的有效驱避剂量差异显著，其中，超临界CO₂萃取物试验所用剂量最小，由初试结果将最高剂量拟定为0.16mg／cm²，其平均的驱避率为87.9％，驱避级别为Ⅴ级，第1天的驱避效果很显著，驱避率为96.6％，到第4天仍有明显的驱避效果；温浸法粗提物效果也较好，当粗提物剂量为0.96mg／cm²～1.6mg／cm²，处理后每天的驱避率都超过了80％，而平均驱避率更是超过了90％，驱避级别均在Ⅴ级。冷浸法、索氏法、冷浸-索氏法得到的粗提物的驱避效果相对较差，因而测试剂量最大，最高剂量设为16mg／cm²，是超临界CO₂萃取物试验测试剂量的100倍。5种方法得到的各粗提物对玉米象都有较好的触杀效果，很小的剂量即可导致玉米象大量死亡。通过对各个时间段所得到的致死中量进行比较，发现超临界CO₂萃取物对玉米象的触杀效果最好，处理玉米象24h、48h、72h后的致死中量分别为0.252mg／cm²、0.232mg／cm²、0.166mg／cm²，与其它方法相比，剂量均为最低。5种方法得到的各粗提物对杂拟谷盗的致死中量从小到大依次为温浸法、SCFE法、冷浸法、冷浸．索氏提取、索氏提取法。SCFE法得到的粗提物对杂拟谷盗的触杀活性除了在前三天的观察中略低于温浸法粗提物，在转入饲料中饲喂一周后，其对杂拟谷盗的LD₅₀与温浸法相当，在处理剂量为0.91mg／cm²时，杂拟谷盗72h后的死亡率达到了90％，在剂量高于0.52mg／cm²时，10d后的死亡率均在80％以上。5种方法得到的各粗提物对谷蠹的致死活性依次为温浸法＞SCFE技术＞冷浸法＞索氏提取法＞冷浸-索氏提取法。温浸法和SCFE法得到的TFG粗提物对谷蠹有良好的触杀活性，处理谷蠹24h后的LD₅₀分别为0.255mg／cm²和0.256mg／cm²，48h后的LD₅₀分别为0.144mg／cm²和0.156mg／cm²，72h后的LD₅₀分别为0.112mg／cm²和0.142mg／cm²，10d后的LD₅₀分别为0.100mg／cm²和0.130mg／cm²。3．以3种溶剂乙醇、乙酸乙酯、氯仿作为夹带剂，对TFG种子干粉进行SCFE，TFG种子粗提物萃取得率由高到低的顺序依次为：乙醇＞乙酸乙酯＞氯仿＞无夹带剂，分别为14.17％、13.31％、10.74％和10.18％。以乙醇作为夹带剂时，所得到的超临界CO₂萃取物对玉米象的触杀活性最强，对玉米象处理24h，48h，72h后的LD₅₀分别为231.764μg／cm²，194.43μg／cm²，141.06μg／cm²；乙酸乙酯作夹带剂时，得到的萃取物对玉米象的触杀效果也较好，对玉米象处理24h，48h，72h后的LD₅₀分别为228.67μg／cm²，180.09μg／cm²和152.61μg／cm²，触杀效果较弱的是氯仿作为超临界CO₂萃取的夹带剂时得到的萃取物，效果最差的是不选用夹带剂，用单一的超临界CO₂作为溶剂而萃取得到的萃取物，处理玉米象72h后的LD₅₀最大，为218.74μg／cm²。不同夹带剂萃取得到的各萃取物对杂拟谷盗成虫的触杀活性的大小依次为氯仿＞乙醇＞无夹带剂＞乙酸乙酯，处理杂拟谷盗10d后的LD₅₀分别为278.57μg／cm²、303.89μg／cm²、341.74μg／cm²和429.45μg／cm²。用乙醇作为夹带剂时得到的超临界CO₂萃取物对谷蠹的触杀效果最好，24h时的LD₅₀为115.14μg／cm²，10d时的LD₅₀为78.25μg／cm²；用单一的超临界CO₂作为溶剂时的萃取物对谷蠹的触杀生物活性最差，24h时的LD₅₀为174．71μg／cm²，10d时的LD₅₀为135.22μg／cm²。4．以萃取率作为目标函数，通过考察分析萃取压力、萃取温度、萃取时间、夹带剂体积和夹带剂浓度对萃取率的影响趋势，确定超临界CO₂萃取过程中各影响因素的正交试验条件为：操作压力的正交试验区间为20MPa～30MPa；萃取温度的正交试验区间为45℃～55℃；正交试验的操作时间为1.5h；正交试验夹带剂用量的区间为20mL～40mL；正交试验夹带剂浓度的区间为60％～95％。选择TFG萃取物的触杀活性作为主要的评判指标，同时以TFG萃取物萃取得率作为补充评判指标，以正交试验对TFG超临界CO₂进行工艺优化，试验结果表明：最佳萃取工艺为萃取压力25Mpa，萃取温度55℃，央带剂为95％乙醇，固液比为200g：30mL。在优化工艺下得到的萃取得率为16.96％，与正交试验方案中的最高值即17.88％基本持平。优化后的SCFE技术工艺得到的萃取物对谷蠹的触杀活性在处理后24h和10d的LD₅₀分别为102.24μg／cm²和65.10μg／cm²，相对毒力分别为1.44和2.26。5．用石油醚、氯仿、乙酸乙酯、正丁醇和水依次对TFG超临界CO₂萃取物（简称TFG-1）进行萃取，萃取率分别为61.32％、9.22％、9.16％、8.39％和17.08％。生物测定结果表明：TFG-1的活性成分主要存在于石油醚萃取物中，对谷蠹成虫表现出很强的触杀作用，TFG-1石油醚萃取物对谷蠹处理24h、48h、72h和10d后的LD₅₀分别为175.05μg／cm²、133.65μg／cm²、113.37μg／cm²和79.34μg／cm²。氯仿萃取物对谷蠹的触杀活性较强，乙酸乙酯萃取物具有一定的活性，而正丁醇萃取物和水提取物未见杀虫活性。以不同极性的溶剂系统作为展开剂，将TFG-1石油醚萃取物进行薄层层析（TLC），在紫外光（254nm）下进行观察，未见明显谱带。说明TFG-1石油醚萃取物没有紫外吸收现象。当以石油醚：乙酸乙酯=10：1的混合溶剂作为展开剂时展层效果最好，在碘显色时有5个谱带，在磷钼酸显色时有10个谱带，其Rf值在0.1-0.9之间。将TFG-1石油醚萃取物进行柱层析，第1次柱层析后，经TLC检测，归类合并，共得到7个流分，第1个组分（Ⅰ）的得率最高，为75.13％，是TFG-1石油醚萃取物的主要成分。当载药量均为519.95μg／cm²时，只有流分Ⅰ的杀虫效果最明显，对谷蠹的致死率在24h时即达到100％，流分Ⅵ其次，对谷蠹的致死率在24h时为80.76％，另外流分Ⅴ也有较好的杀虫效果，对谷蠹的致死率在24h时为46.67％，而流分Ⅱ、Ⅳ和Ⅶ的杀虫效果较差，流分Ⅲ几乎没有杀虫效果。流分Ⅰ柱层析分离得到5个样品，分别为样品a、样品b、样品c、样品d、样品e，其中样品e为流分Ⅰ的主要成分。流分Ⅵ柱层析分离后得到4个样品，分别为样品f、样品g、样品h、样品i，其中样品i为流分Ⅵ的主要成分。生物测试结果显示从流分Ⅰ中得到的5个样品均有杀虫活性，样品e对谷蠹成虫的触杀活性很强，当处理剂量为259.98μg／cm²时，谷蠹经样品e处理后，其24h时的死亡率达77.78％。从流分Ⅵ中得到的4个样品同样均有杀虫活性，样品i的触杀活性较强，谷蠹经剂量为259.98μg／cm²的样品i处理，其24h后的死亡率为41.11％。样品e经多次柱层析纯化得到一单体化合物e5222（简称化合物Ⅰ）。样品i经柱层析后，在丙酮中重结晶，得到一结晶状单体化合物i3111（简称化合物Ⅱ）。经理化常数测定和红外、核磁共振、质谱等波谱数据分析鉴定，化合物Ⅰ和化合物Ⅱ分别是1，2-二亚油酰-3-硬脂酰-甘油三酯（1，2-dilinoleic acid-3-stearic acid-triglyceride）和4a，14a-二甲基-胆甾醇（4a，14a-dimethyl-cholesterol），这2个化合物均首次从TFG中发现。6．化合物Ⅰ对谷蠹成虫有较强的触杀活性，剂量为129.99μg／cm²的药剂处理试虫24h、48h、72h和10天后，谷蠹的平均死亡率分别为32.22％、40.00％、56.67％和70.00％，死亡率较高。化合物Ⅰ对玉米象有明显的触杀效果，用剂量为129.99μg／cm²的药剂处理试虫24h、48h、72h和10天后，玉米象的平均死亡率分别为86.67％、92.22％、95.56％和100.00％，死亡率很高。化合物Ⅱ对谷蠹和玉米象的触杀效果相对较弱，当药膜剂量为259.98μg／cm²时，对谷蠹处理10天后有少量成虫死亡，平均死亡率为4.44％，同样的剂量处理玉米象10天后的平均死亡率为97.78％。将化合物Ⅰ与化合物Ⅱ按质量比为1：1制成混剂，对谷蠹和玉米象的生测结果表明：处理谷蠹成虫10天后，得到的共毒系数为60.57，共毒系数数值小于80，两单体的活性在处理谷蠹成虫10天后表现为拮抗作用，混剂处理玉米象10天后，得到的共毒系数为30.23，共毒系数数值小于80，两单体对玉米象的活性同样表现为拮抗作用。更多还原

【Abstract】 Aim to study the bioactivity against the main stored pests including Sitophilus zeamais Motschulsk （S. zeamais）, Rhyzopertha dominica Fabricius （R dominica）, Tribolium castaneum （Herbst） （T.castaneum） and Oryzaephilus surinamensis （Linnaeus）（O. surinamensis）, several solvents and methods were selected to extract the bioactive substance efficiently from Legumiosae plant TFG seeds. From the year of 2003 to 2006, a series of studies concerned plant TFG have been conducted including its contact toxicity, repellency and its phytochemistry, the research results are outlined as follows:1. The powderded seeds of TFG was extracted with alcohol, chloroform, ethyl acetate, acetone and petroleum ether at relative high temperature by dipping the extract flask in warm water, the extracts rate of TFG seeds in alcohol was 11.48%, which is the highest rate among these solvents, while petroleum ether had the lowest extract rate which was only 4.91%. The extracts rate of TFG in chloroform, ethyl acetate and acetone was 7.24%, 7.54% and 5.29%.The five extracts obtained from different solvents all had a certain repellency against S. zeamais, the chloroform extract had the most effective repellency among those extracts, and its repellency level against S. zeamais could reach toⅡwhen dosage was at 1000μg/cm², the second was acetone extracts.All the extracts had a significant repellency to T.castaneum, average repellency of different extracts all exceeded 80%, and repellency level wereⅤ, acetone extracts had the most effective repellency to T.castaneum. the second one was chloroform extracts, the one had the most poor repellency was ethyl acetate. The difference of repellency rates between S. zeamais and O. surinamensis did not caused obviously by TFG extracts, while the repellency rate between S. zeamais and T.castaneum was notable, and the repellency rate between R. dominica and O. surinamensis was also notable. The order of repellency against stored pests by TFG solvent extracts was T.castaneum＞S. zeamais＞O. surinamensis＞R. dominica.All the five extracts possessed strong contact activity to S. zeamais, the adults, treated by extracts for three days and then cultured by clean wheat for one week, nearly all died. The alcohol extracts had the most effective contact toxicity to S. zeamais, its LD₅₀ （effective dosage causing 50% mortality rate）to S. zeamais was 0.362mg/cm² and 0.328mg/cm², respectively. The second was ethyl acetate extracts and its LD₅₀ to S. zeamais within 48 hours was 0.334mg/cm², acetone extracts also had excellent contact toxicity to S. zeamais and its LD₅₀ within 24 hours to S. zeamais was 0.380mg/cm².The petroleum ether extract had the lowest contact toxicity to S. zeamais among the five TFG extracts. The LD₅₀ of petroleum ether extract to S. zeamais was 0.772 mg/cm² and 0.680 mg/cm² by treatment within 24 hours and 48 hours, respectively.All the TFG five extracts had poor contact toxicity to T. castaneum adults. The mortality of T. castaneum changed very little after treated for three days and then cultured for one week. TFG alcohol extracts showed the most effective activity to R. dominica among these extracts, its LD₅₀ to R. dominica adults was 0.482 mg/cm², 0.450mg/cm²,0.400mg/cm² and 0.386mg/cm² within 24 hours, 48 hours, 72 hours and 10 days; the TFG petroleum ether extracts had the hightest LD₅₀ to T. castaneum.TFG actone extract had strong toxicity to R. dominica with the LD₅₀ of 0.240mg/cm² by treatment within 24 hours, it showed more efficient than alcohol extract and chloroform extract, the mortality of R. dominica adults increased rapidly from 24 hours to 48 hours after treatment. The mortality of O. surinamensis changed differently according to the different TFG extracts and treatment dosage, alcohol extracts and chloroform extracts had more active contact toxicity than other extracts.2. Since alcohol was the optimum solvent to extract bioactive substance according to the results of the first experiment, it was selected as extract solvent for different extracting method including method of dipping powered TFG in alcohol at room temperature （TFGR）, method of dipping powdered TFG in alcohol at relative high temperature （TFGH）, method of Soxhlet extraction （TFGS）, method of TFGR and TFGS （first, dipping powered TFG in alcohol at room temperature to get some extracts, then extracting remainder with Soxhlet extraction apparatus to obtain more extracts, TFGR-TFGS）, method of Supercritical CO₂ Fluid Extraction （TFG-SCFE）, no modifier was used in TFG-SCFE. The results showed that the extracts rate of TFGR, TFGH, TFGS, TFGR-TFGS, TFG-SCFE was 4.65%, 11.48%, 12.99%, 8.14%, 11.35%, respectively.All the extracts obtained from those five methods were good at repelling T.castaneum, the repellency rates varied obviously when the extract method changed. TFG-SCFE extracts could repel T.castaneum at very little dosage. Treatment with TFG-SCFE extracts at dosage 0.16 mg/cm², the average repellency rate within 4 days was 87.9%; treatment with TFGH extracts at dosage about 0.96 mg/cm² to 1.60 mg/cm², the average repellency rates of everyday all exceeded 80%, and the average repellency rates within 4 days was high to more than 90%, TFGR extracts, TFGS extracts and TFGR- TFGS extracts all had relative poor repellency to T.castaneum, the highest dosage for testing was 16 mg/cm², which was 100 times to the testing dosage of TFG-SCFE extracts.The LD₅₀ values of extracts obtained from those five methods to T.castaneum increased gradually from TFGH, TFG-SCFE, TFGR, TFGR-TFGS and TFGS. TFG-SCFE extracts had similar contact toxicity as TFGH extracts, treatment with it at dosage 0.91 mg/cm² on T.castaneum, the mortality achieved to 90% within 72 hours, and the mortality all exceeded 80% when dosage above 0.52 mg/cm².The toxicity order to R. dominica was TFGH＞TFG-SCFE＞TFGR＞TFGS＞TFGR-TFGS. TFGH extracts and TFG-SCFE extracts showed effective contact toxicity to R. dominica, the LD₅₀ of TFGH to R. dominica within 24 hours, 48 hours,72 hours and 10 days was 0.255 mg/cm², 0.144 mg/cm², 0.112mg/cm² and 0.100mg/cm², respectively. The LD₅₀ of TFG-SCFE to R. dominica within 24 hours, 48 hours,72 hours and 10 days was 0.256 mg/cm², 0.156 mg/cm², 0.142mg/cm² and 0.130mg/cm², respectively.3. Alcohol, ethyl acetate, chloroform was used as modifier to extract bioactive substance from TFG seeds powder, and the extract rates of TFG-SCFE with alcohol （TFG-SCFE-A）, TFG-SCFE with ethyl acetate （TFG-SCFE-E）, TFG-SCFE with chloroform（TFG-SCFE-C）, and TFG-SCFE without modifier was 14.17%,13.31%,10.74% and 10.18%. Among those four extracts, TFG-SCFE-A extracts was the best pesticide to kill 5. zeamais. The LD₅₀ of TFG-SCFE-A to 5. zeamais within 24 hours, 48 hours, 72 hours was 231.76μg/cm², 194.43 mg/cm², 141.06μg/cm² respectively. The second was TFG-SCFE-E extract, its LD₅₀ to S. zeamais within 24 hours, 48 hours and 72 hours was 228.67μg/cm²,180.09μg/cm² and 152.61μg/cm², the third was TFG-SCFE-C, and the last one was TFG-SCFE, the LD₅₀ of TFG-SCFE to S. zeamais within 72 hours was the largest one, and the value was 218.74μg/cm².The contact toxicity order of the SCFE extracts with different modifier against T. castaneum was TFG-SCFE-C＞TFG-SCFE-A＞TFG-SCFE＞TFG-SCFE-E, the LD₅₀ of them to T.castaneum was 278.57μg/cm²,303.89μg/cm²,314.74μg/cm² and 429.45μg/cm².Among these four extracts, TFG-SCFE-A extracts possessed strong pesticidal effect to R. dominica, the LD₅₀ of it to R. dominica within 24 hours and 10 days was 115.14μg/cm² and 78.25μg/cm², respectively; TFG-SCFE extracts was the one that showed poor contact toxicity, the LD₅₀ of it to R. dominica within 24 hours and 10 days was 174.71μg/cm² and 135.22μg/cm².4. Used extract rate as standard values, extract pressure （EP）, extract temperature （ET）, extract duration （ED）, modifier volume （MV）, modifier consistence （MC） were investigated to settle down optima range, the results showed range for EP was 20MPa～30MPa, range for ET was 45℃～55℃, range for ED was 1h～2h, range for MV was 20mL～40mL and range for MC was 60%～95%.Contact toxicity and extract rate were used as standard values to optimize extraction conditions, which were achieved at 55 ^ΩC under 25 Mpa of pressure. The extraction efficiency for 200 grams of dry sample reached 16.96% with 30 mL of 95% alcohol. Extracts loaded on filter paper showed dose and time dependent toxicities to adult R. dominica with a LD₅₀ value of 65.10μg/cm² after 3d treatment and 7d culturation.5. TFG-1 extract was first separated into five parts by partition in liquid extraction method, petroleum ether, chloroform, ethyl acetate, n-butanol and water were used, extract rate of them was 61.32%,9.22%,9.16%,8.39% and 17.08%. the petroleum ether fraction was identified to be the most active part as result of contact toxicity; the second, chloroform fraction, and the third, ethyl acetate, treatment with TFG-1 n-butanol extracts and TFG-1 water extracts to R. dominica showed no contact toxicity. and the LD₅₀ of TFG-1 petroleum ether extract to R. dominica within 24 hours, 48 hours, 72 hours and 10 days was 175.05μg/cm²,133.65μg/cm²,113.37μg/cm² and 79.34μg/cm², respectively.Separation and purification of the petroleum ether part was performed through silica gel column for several times. Seven parts were got by TLC testing, the separation rate of partⅠwas the highest one, and the value was 75.13%, which mean partⅠwas the main ingredients of TFG-1 petroleum ether extract. Treatment with extracts of partⅠ, partⅥand partⅤat dosage 519.95μg/cm² on R. dominica, the mortality rate of adults was 100%, 80.76% and 46.67% within 24 hours, partⅡ, partⅣand partⅦhad little contact toxicity to R. dominica, partⅢshowed no toxicity to R. dominica by those testing treatment dosages. Five samples were obtained from partⅠby column chromatography, that was sample a, sample b, sample c, sample d and sample e, and results of separation rates showed sample e was the main components of partⅠ. Four samples were got from partⅥ,and sample i was the main component of partⅥ. Treatment with sample e and sample i at dosage 259.98μg/cm² to R. dominica, the mortality of adults was 77.78% and 41.11%, respectively.One monomer compound （compoundⅠ）was isolated from the active sample e, the molecular structure of the compound was definitely identified as 1,2-dilinoleic acid-3-stearic acid-triglyceride by the spectrum analysis including MS, ¹H-NMR, ¹³C-NMR and IR. One crystal （compoundⅡ） was isolated from sampleⅠ, and was named as 4a, 14a-dimethyl-cholesterol.6. It was found that compoundⅠpossessed strong contact toxicity to R. dominica and S. zeamais, treatment with dosage 129.99μg/cm², the average mortality of R. dominica and S. zeamais within 10 days was 70.00% and 100.00%, respectively, compoundⅡalso showed contact toxicity to R. dominica and S. zeamais, treatment with dosage 259.98μg/cm², the average mortality of R. dominica and S. zeamais within 10 days was 4.44% and 97.78%. After having mixed compoundⅠand compoundⅡat the weight rate 1:1, the mixture was tested its contact toxicity to R. dominica and S. zeamais. The results showed treatments of mixed compound had pesticidal effects on R. dominica and S. zeamais, whose LD₅₀ were 217.60μg/cm² and 45.44μg/cm² within 10 days; the co-toxicity coefficient of mixed medicament to R. dominica and S. zeamais were 60.57 and 30.23, the values were much smaller than 80, so their medicamental avail relaxational effect.更多还原