杜香精油杀虫活性物质基础、构效关系及其变化规律研究

Ledum palustre is an important perfume plant in northeast China, and widely distributed in Daxing’anling area. At least since the eighteenth century it has been used in ethnomedicine for the treatment of various ailments, such as rheumatism, cough, cold and insect bites, as well as a repellent. The essential oil of wild L. palustre with the rich polyphenolic fraction possesses analgesic, anti-inflammatory, antimicrobial, antiviral, antifungal and insecticidal potential, demonstrated by in vivo and in vitro studies. The components and contents of essential oils in L. palustre were highly correlated with their growth environment. The most present research on L. palustre essential oil is still focused on the analysis of the essential oil composition. The insecticidal active compounds of essential oil and their variability regularity from wild L. palustre were not very clear as yet. Our objective is to investigate the insecticidal active compounds of essential oil and their diurnal and mensal variation from wild L. palustre in Daxing’anling area. The main flavor compounds in L. palustre leaves, twigs and stems were isolated by sub-critical fluid extraction technology and simultaneous distillation and extraction (SDE), and then were qualitatively and quantitatively analyzed by gas chromatography-mass spectrometry (GC-MS). In order to further study the insecticidal components in essential oil of L. palustre, bioactivity-directed fractionation and purification processes were employed to identify the insecticidal components by silica gel column chromatography. The chemical structures of these insecticidal components were identified by IR, MS, UV, and NMR. The best formula of complex natural insecticide was screened by mixed compounds from L. palustre essential oil. The insecticidal activity and structure-activity relationship of L. palustre essential oil were studied. The diurnal and mensal variation of these insecticidal components in essential oil of L. palustre was studied using GC-MS and HS-SPME-GC-MS analysis. This work will provide the fundamental theories for development and utilization of resource of L. palustre in Daxing’anling area. For developing new efficient green plant source agent of insecticide provided theoretical basis.

杜香是我国大兴安岭地区蓄积量丰富的野生植物资源，杜香精油对仓储害虫具有熏蒸活性，但其杀虫活性物质基础及其变化规律目前尚不清楚。本项目拟以大兴安岭地区杜香为材料，通过亚临界萃取和同时蒸馏萃取技术对杜香精油组分进行提取分离；采用生物活性追踪法、硅胶柱层析技术，从杜香精油中分离、纯化有效熏蒸杀虫物质；通过现代波谱学技术对分离得到的杀虫活性物质进行解析，阐明其化学结构；通过对杜香精油不同单体成分的复配模拟，建立杜香精油组分熏蒸杀虫效果的量化模型，研究复配精油组分对不同种类仓储害虫的熏蒸杀虫效果，筛选出最佳配方；探索研究杜香精油组分的化学结构、浓度对熏蒸杀虫效果影响的内在规律；利用SPME-GC-MS技术对昼夜不同时间及不同月份的杜香精油中有效杀虫物质及其含量进行测定，以期探明昼夜变化及不同月份杀虫有效成分的变化规律。本研究将为合理、高效开发利用杜香资源，研发新的环境友好熏蒸剂提供理论基础。

杜香是我国大兴安岭地区蓄积量丰富的野生植物资源，杜香含有丰富的挥发性芳香油，是极具利用潜力的资源植物。杜香精油含量和组分是衡量杜香品质的最重要指标。国外有学者对生长在欧洲不同地域的杜香精油含量和组分进行过研究，发现杜香精油含量和成分因亚种、变种、品种、栽培地的环境条件、部位及其发育阶段而异。精油成分的相对含量不同，直接影响精油芳香气味，进而影响精油的商品价值。目前，国内外对杜香精油的研究大部分还停留在对杜香精油成分的分析上，但对杜香精油杀虫等有效物质基础及其构效关系尚不清除，对杀虫有效成分在杜香生长发育过程中的变化规律尚不明确。本项目拟对我国大兴安岭地区杜香精油杀虫活性物质基础、构效关系及其含量变化规律进行系统的研究。本项目分别完成了利用水蒸气蒸馏法、溶剂提取法及亚临界萃取技术对杜香枝叶中挥发性成分的提取试验，确定了杜香枝叶活性成分的最佳提取工艺；对其所得活性成分进行了分析鉴定；杜香枝叶提取物经GC-MS测定，其中叶挥发油含有40多种组分，枝挥发油含有近30种组分，主要以α-蒎烯、β-蒎烯、α-侧柏烯、β-侧柏烯、桧稀、苧烯、对伞花烃、桃金娘烯醛、莳萝醛为主；完成了不同采集时间的杜香枝叶经水蒸气蒸馏提取得到的精油对四种储粮害虫的熏蒸活性和触杀活性；结果显示杜香精油熏蒸处理48 h后，赤拟谷盗、谷蠹、玉米象和烟草甲四种储粮害虫具有良好的杀虫效果；本项目已申请国家发明专利2项，发表研究论文5篇（其中SCI收录3篇），申请软件著作权1项。本项目研究内容尚未见其他相关报道，属于申请者原创性研究工作。本项目的研究为研发新的环境友好熏蒸剂提供理论基础，对我国杜香资源的开发利用和产业化生产具有重要的意义。