【作者】 宋兵；

【导师】 韩崇选；

【作者基本信息】 西北农林科技大学 ， 森林保护， 2009， 硕士

【摘要】 本论文主要对苦参中的氧化苦参碱、槐定碱、苦参碱三种生物碱作为活性成分开展植物源灭鼠药剂研制进行了系统的研究。通过适口性试验、三者间相互作用关系测定、对小鼠的亚急性毒性试验以及应用于鼠药研发的微囊制作初试,得出以下结果:1.小鼠对添加了不同含量的氧化苦参碱、槐定碱的饵料摄食系数较之CK组都有降低,拒食现象明显;对不同含量的苦参碱饵料表现为较好的适口性,当含量为0.1%时,摄食系数甚至高于CK组,对小鼠的适口性有促进作用。三者对小鼠体重的影响显著,即随着浓度的增大,体重降低越明显。2.从三种苦参碱和溴敌隆混合后的毒性结果来看,其-20<c.f <20范围内,表现为相加作用。表明了3种苦参生物碱与溴敌隆混合后对小鼠的毒性增加不显著。而摄食系数较之前期的适口性测定结果有明显的提高。3.混合后的三种苦参生物碱毒性增加,苦参碱+槐定碱的增效作用最强,协同毒力指数(c.f)值达到59.1,氧化苦参碱+苦参碱混合给药,c.f值为37.3,表现为增效作用;氧化苦参碱+槐定碱混合给药,c.f值为13.6,表现为相加作用。4.亚急性毒性试验结果表明:血常规检测结果确定三种苦参生物碱处理组小鼠的白细胞计数(WBC)、血小板总数(PLT)、中粒细胞(GR)较之对照组显著增高;血液生化指标检测结果表明槐定碱和氧化苦参碱各处理组小鼠的尿素氮(BUN)、肌酐(Cr)水平均高于对照组(p<0.01),苦参碱各处理组小鼠的丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基酸转移酶(AST)水平较之对照组升高(p<0.01);脏器系数的统计结果表明:槐定碱、氧化苦参碱各处理组小鼠的肾脏系数、心脏系数以及肝脏系数均显著地高于对照组(p<0.01),苦参碱处理组小鼠的肾脏系数、心脏系数及肺脏系数显著地高于对照组(p<0.01、p<0.05、p<0.05)。由此说明以上三种苦参生物碱对肝脏、肾脏有影响,初步确定他们作用的靶器官是肝脏和肾脏。5.采用正交试验设计确定最佳的制备苦参生物碱微囊的方法,结果表明当氧化苦参碱微囊囊心囊材比为1.00:0.75,转速为600rpm,温度为70℃时含量较高;当槐定碱微囊的囊心囊材比为0.75:0.75,转速为600rpm,温度为50℃时生物碱含量较高;囊心囊材比为1.00:0.75,转速为600rpm,温度为70℃时制得的苦参碱微囊所含的生物碱含量高。本实验利用改良后的工艺制备苦参生物碱微囊,所得微囊是颗粒状,流动性好。6.通过紫外分光光度计测定,三种苦参生物碱分别以414nm、411nm、413nm为检测波长,吸光度A为纵坐标,含量B为横坐标,得出苦参碱的线性回归方程为:A=0.0104B+0.0084 R~2=0.9994 ( n=7 );氧化苦参碱的线性回归方程为:A=0.0103B-0.0191 R~2=0.999 ( n=7 );槐定碱的回归方程为: A=0.0111B-0.0164 R~2=0.9991(n=7),且三者在0—100μg范围内线性关系良好。通过加样回收率试验表明:三种苦参生物碱微囊的加样回收率较高,苦参碱的为91.59%;槐定碱微囊的为97.72%;氧化苦参碱微囊的为103.65%。所制备的微囊在24h内有较好的稳定性。苦参碱微囊样品中生物碱含量为15.0%;氧化苦参碱微囊样品中生物碱含量为16.7%;槐定碱微囊样品中生物碱含量为18.5%。7.投食试验结果表明,添加了苦参碱微囊的饵料对小鼠的适口性有较大的提高,很好地改善了小鼠拒食的现象。试鼠体重没有明显的下降,呈平稳状况。由小鼠的死亡数量来看,死亡率为25%,其毒性有待进一步提高。更多还原

【Abstract】 The toxicity of three Sophora flavescens Ait alkaloids (Oxymatrine, Sophoridine, Matrine) as active components in botanical rodenticide had been systematically researched. Tests of palatability, interactions between three alkaloids and subacute toxicities to mice, as well as preliminary experiments on microcapsules used in rodenticide, came to the following conclusions:1.Feed coefficient of mice in which groups baits were added different content of Oxymatrineor Sophoridine was lower than that of the CK group, and the antifeedant was obvious. Bait added Matrine had better peformances on palatability, especially when the content is 0.1%, the feed coefficient is even higher than the CK group. The body weights of mice decreases phenomenally as the content of alkaloids increases.2.A c.f value ranging from -20 to 20 calculated with the mixture-toxicity of Bromadiolone and either of three Sophora flavescens Ait alkaloids showed an additive effect on toxicity. The mixture-toxicity increases unobviously, however the feed coefficients increase phenomenally.3.Mixtures of either two of three Sophora flavescens Ait alkaloids have higher toxicities. Mixtures of Matrine and Sophoridine have the greatest synergized action on toxicity with a c.f value equal to 59.1. Mixtures of Oxymatrine and Matrine have synergized action on toxicity with a c.f value equal to 37.3. Mixtures of Oxymatrine and Sophoridine have additive effect on toxicity with a c.f value equal to 13.6.4.Results of subacute toxicity tests showed: In blood routine the WBC,PLT,GR of mice in Sophora flavescens Ait alkaloid-added groups were higher than CK groups. Blood biochemical index indicates that BUN, Cr level of Sophoridine group and Oxymatrine group were both higher than CK group(p<0.01), while ALT, AST level of Matrine group were higher than CK group. The coefficients of kidney, heart and liver in Sophoridine and Oxymatrine groups were significantly higher than that of CK groups(p<0.01), while the coefficients of kidney, heart and lungs in Matrine group were significantly higher than that of CK group(p<0.01、p<0.05、p<0.05). According to these above,the target organs on which Sophora flavescens Ait alkaloids acted were kidney and liver.5.Orthogonal experiments were designed to determine the optimal microcapsule preparation method of Sophora flavescens Ait alkaloids. Results showed that content of Oxymatrine in microcapsules were at higher level under a rotation speed of 600rpm and a temperature of 70℃when the microcapsule core-wall ratio was 1.00:0.75. The content of Sophoridine were at higher level under a rotation speed of 600rpm and a temperature of 50℃when the core-wall ratio was 0.75:0.75. The content of Matrine were at higher level under a rotation of 600rpm and a temperature of 70℃. Microcapsules made by refined technology in this experiment was granulated and of good flow ability.6.Measured by UV spectrophotometer, detection wavelength of three alkaloids were 414nm,411nm and 413nm respectively. Absorbance (A) as the longitudinal coordinates and content (B) as the abscissa, a linear regression equation for Matrine was calculated to be: A=0.0104B+0.0084 R~2=0.9994(n=7); for Oxymatrine the linear regression equation was: A = 0.0103B-0.0191 R~2 = 0.999 (n = 7); for Sophoridine the linear regression equation was: A = 0.0111B-0.0164 R~2 = 0.9991 (n = 7). These three have a good linear relationship in the alkaloid content range of 0-100μg. Recovery of Sophora flavescens Ait alkaloids from microcapsules showed that: the three alkaloids microcapsules have higher recovery rate,91.59% for Matrine, 97.72% for Sophoridine, and 103.65% for Oxymatrine. Microcapsules had a better stability in 24h. The alkaloid content of Matrine microcapsule sample was 15.0%, while Oxymatrine microcapsule sample was 16.7% and Sophoridine microcapsule sample was 18.5%.7.Baits for mice added Sophora flavescens Ait alkaloids microcapsules had a better palatability and reduced the antifeedant rate. There was no obvious decrease in mouse weights but a stable state. The death rate was 25%, which indicated that the toxicity of microcapsule needed a further improvement.更多还原