CN106259312B - 一种植物精油-低聚壳聚糖复合抗植物真菌微乳剂 - Google Patents
一种植物精油-低聚壳聚糖复合抗植物真菌微乳剂 Download PDFInfo
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Abstract
本发明公开了一种植物精油‑低聚壳聚糖复合抗植物真菌微乳剂。该抗植物真菌微乳剂按重量百分比计由以下原料经乳化、复配而成:植物精油5‑30%、乳化剂15‑30%、助溶剂5‑10%、低聚壳聚糖0.5‑2%、石榴皮提取物0.5‑1%,余量为水。本发明以抗菌活性较强的艾油、茶树油及低聚壳聚糖和抗氧化活性优越的石榴皮提取物为原料,通过乳化、复配得到植物精油‑低聚壳聚糖复合抗植物真菌微乳剂,主要用于防治苹果、樱桃等果树的轮纹病、褐腐病、炭疽病、灰霉病等,及苹果、樱桃等水果的保鲜。
Description
技术领域
本发明涉及一种植物精油-低聚壳聚糖复合抗植物真菌微乳剂,属于农药技术领域。
背景技术
随着化学农药及化学食品添加剂引发的食品安全问题和对环境危害的不断加剧,以及消费者对有机食品的日益认可,生物源农药和添加剂的开发使用越来越受到广大科研工作者的重视。
植物精油属于植物体内的次生代谢物质,由于具有较强的抑菌活性和低毒、环境友好、气味芳香等特点,近年来被广泛开发用于水果、食品保鲜及果蔬植物抗菌等。
其中,艾油是由菊科植物艾提取的挥发油,为淡黄色或黄绿色澄明液体,具有平喘、镇咳、祛痰、消炎、抗过敏等药效。近年来研究发现,它对小麦赤霉病菌、苹果轮纹病菌、苹果斑点落叶病菌、桃褐腐菌、苹果炭疽病菌、辣椒炭疽病菌、柑橘炭疽病菌和番茄早疫病菌等十三种植物病原菌均有抑制作用。茶树油是由桃金娘科白千层属灌木新鲜枝叶蒸馏得到的芳香精油,具有广谱的抗菌、抗真菌能力和抗氧化能力。中国卫生部于2003年已将茶树油列入食品香料名单。最新研究发现,茶树油对番茄枯萎灰霉菌、灰葡萄胞霉及引起采摘后水果腐烂的炭疽病菌有明显的抑制作用。
总之,植物精油在抗菌方面具有广谱、高效、起效快的优势,但由于植物精油本身易挥发、不溶于水,必须制备成合适的剂型后使用以保持其持效性。
基于以上现有技术,由“北京市教育委员会市属高校创新能力提升计划项目”资助完成本发明。
发明内容
本发明的目的是提供一种植物精油-低聚壳聚糖复合抗植物真菌微乳剂,主要用于防治苹果、樱桃等果树的轮纹病、褐腐病、炭疽病、灰霉病等,及苹果、樱桃等水果的保鲜。
为实现上述目的,本发明采用以下技术方案:
一种植物精油-低聚壳聚糖复合抗植物真菌微乳剂,按重量百分比计由以下原料经乳化、复配而成:植物精油5-30%、乳化剂15-30%、助溶剂5-10%、低聚壳聚糖0.5-2%、石榴皮提取物0.5-1%,余量为水。
所述植物精油为艾油和茶树油中的一种或两者的混合物。所述乳化剂为吐温-80、OP-10、Turmul5030、AEO-5中的一种或多种。所述助溶剂为聚乙二醇、聚乙烯醇、丙三醇中的一种。所述低聚壳聚糖为相对分子质量低于5kDa的低聚壳聚糖。所述石榴皮提取物需要经过微滤膜纯化处理。
所述植物精油-低聚壳聚糖复合抗植物真菌微乳剂的制备方法为:首先按质量配比准备植物精油、乳化剂和助溶剂,在室温下混合搅拌均匀;在搅拌的条件下继续向上述体系中加入一定质量百分比浓度的低聚壳聚糖水溶液和一定质量百分比浓度、经过微滤纯化处理的石榴皮提取物水溶液;再进行高速剪切、搅拌至澄清状,即获得微乳剂。
本发明的优点在于:
1、本发明的抗植物真菌微乳剂不仅具备微乳剂分散度极高、对靶体渗透性强、附着力好等优势,还直接以抗菌效果优异的艾油和茶树油作为油相,不使用任何有机溶剂,与常见乳油农药相比可大大减轻有机溶剂对环境造成的压力。
2、本发明利用抗菌活性较强的艾油、茶树油和低聚壳聚糖的抑菌协同性,利用抗氧化活性优越的石榴皮提取物的抗氧化特性,从抗菌和提高抗逆性两个方面缓解果树的病害;从抗菌和抗氧化两个方面来增强果蔬的保鲜效果。此外,分子量相对较大的低聚壳聚糖和乳化剂的吸附作用可减缓植物精油的挥发。同时,具有较强抑菌活性低聚壳聚糖的存在也保证了制剂的抗菌持效性。
3、本发明中,艾油、茶树油、低聚壳聚糖和石榴皮提取物经过乳化、复配,克服了艾油和茶树油不溶于水、而艾油、茶树油、低聚壳聚糖和石榴皮提取物不能互溶的缺点,形成可与水任意互溶的植物精油-低聚壳聚糖微乳剂,降低了实际使用的难度。
具体实施方式
以下通过具体实施例对本发明做进一步说明。
石榴皮中含有多酚、黄酮及生物碱等次生代谢产物,具有显著的抗菌和抗氧化生物活性。研究表明,石榴皮提取物稀释100倍后对番茄灰霉病菌的防治效果达到了46.4%;其清除自由基及抑制脂质过氧化的能力强于没食子酸和绿茶多酚。可通过提高抗氧化能力和清除水果内氧自由基来有效防止水果衰老和品质下降。
壳聚糖是虾、蟹及昆虫外壳中的甲壳素脱乙酰基后的产物,不仅具有良好的生物相溶性、成膜性、保湿性和可生物降解性,还可提高农产品的品质。研究表明,相对分子量低于5000的低聚壳聚糖及其衍生物表现出较强的抗菌活性,壳聚糖用于农药载体和水果保鲜剂的开发多有报道。
实施例1
各组分配比约为:茶树油5份,OP-10 30份,聚乙二醇4000 5份,相对分子质量4kDa的低聚壳聚糖2份,石榴皮水提物0.5份,水57.5份。
具体实施步骤为:向洁净干燥的容器中按次序加入300g OP-10,50g聚乙二醇4000,50g茶树油,添加完毕后,用机械搅拌器搅拌均匀。在搅拌条件下,向上述体系中继续加入质量百分比含量为7.0%、相对分子质量为4kDa的低聚壳聚糖290mL和经过微滤纯化处理、质量百分比含量为1.7%的石榴皮提取物水溶液290mL。混合均匀后将上述配制的溶液倒入乳化机内,在3000r/p条件下乳化3min,所得体系为澄清状微乳液。
实施例2
各组分配比约为:艾油2.5份,茶树油2.5份,吐温80 25份,丙三醇10份,相对分子质量2kDa的低聚壳聚糖0.5份,石榴皮水提物1份,水58.5份。
具体实施步骤为:向洁净干燥的容器中按次序加入250g吐温80,100g丙三醇,25g艾油和25g茶树油,添加完毕后,用机械搅拌器搅拌均匀。在搅拌条件下,向上述体系中继续加入质量百分比含量为1.7%、相对分子质量为2kDa的低聚壳聚糖溶液300mL和经过微滤纯化处理、质量百分比含量为3.5%的石榴皮提取物水溶液300mL。混合均匀后将上述配制的溶液倒入乳化机内,在3000r/p条件下乳化3min,所得体系为澄清状微乳液。
实施例3
各组分配比约为:茶树油5份,Turmul5030 15份,聚乙烯醇5份,相对分子质量2kDa的低聚壳聚糖2份,石榴皮水提物1.0份,水77份。
具体实施步骤为:向洁净干燥的容器中按次序加入150g Turmul5030,50g聚乙烯醇,50g茶树油,添加完毕后,用机械搅拌器搅拌均匀。在搅拌条件下,向上述体系中继续加入质量百分比含量为5.6%、相对分子质量为2kDa的低聚壳聚糖水溶液340mL和经过微滤纯化处理、质量百分比含量为2.8%的石榴皮提取物水溶液330mL。混合均匀后将上述配制的溶液倒入乳化机内,在3000r/p条件下乳化3min,所得体系为澄清状微乳液。
实施例4
各组分配比约为:艾油1份,茶树油4份,AEO-5 20份,丙二醇10份,相对分子质量4kDa的低聚壳聚糖1.5份,石榴皮水提物0.5份,水63份。
具体实施步骤为:向洁净干燥的容器中按次序加入200gAEO-5,100g丙二醇,10g艾油和40g茶树油,添加完毕后,用机械搅拌器搅拌均匀。在搅拌条件下,向上述体系中继续加入质量百分比含量为5.0%、相对分子质量为4kDa的低聚壳聚糖水溶液320mL和经过微滤纯化处理、质量百分比含量为1.6%的石榴皮提取物水溶液320mL。混合均匀后将上述配制的溶液倒入乳化机内,在3000r/min条件下乳化3min,所得体系为澄清状微乳液。
实施例1-4中所制备植物精油-低聚壳聚糖复合微乳剂性能测定
(1)粒径分布测定
利用激光粒度仪对实施例1-4中所制备植物精油-低聚壳聚糖复合微乳剂进行粒径测定,结果显示:实施例1-4中所制备的植物精油-低聚壳聚糖复合微乳剂粒径均在0.001-0.1um之间,符合微乳剂的粒径要求。
(2)稳定性测定
参照农药乳液稳定性测定方法(GB-T1603-2001),用标准硬水分别稀释实施例1-4中所制备的植物精油-低聚壳聚糖复合微乳剂,水浴(30±2℃)条件下放置1h,观察四种样品的表观现象。
参照农药热贮稳定性测定方法(GB-T 19136-2003),分别将实施例1-4中所制备的植物精油-低聚壳聚糖复合微乳剂,密封,在54±2℃条件下放置14d,观察四种样品的表观现象。
参照农药低温稳定性测定方法(GB-T 19137-2003),分别将实施例1-4中所制备的植物精油-低聚壳聚糖复合微乳剂在0±1℃条件下放置7d,温度恢复至室温,观察四种样品的表观现象。
结果发现,实施例1-4中所制备的复合微乳剂经过以上三组实验,均没有分层、浮油和结晶或沉淀等现象产生。说明所制备的四种微乳剂稳定性合格。
(3)对苹果轮纹病菌、桃褐腐病菌的抑制活性测定
分别以茶树油乳剂、艾油乳剂以及相对分子量为4kDa低聚壳聚糖水溶液作为对照组,采用稀释法,研究实施例1-4中所制备植物精油-低聚壳聚糖复合微乳剂及茶树油乳剂、艾油乳剂和低聚壳聚糖水溶液对苹果轮纹病菌和桃褐腐病菌的抑制现象。
其中茶树油乳剂各组分配比为:5份茶树油、20份OP-10、8份丙三醇,67份水;艾油乳剂各组分配比为:5份艾油、22份吐温-80、10份丙三醇,63份水;相对分子量为4kDa低聚壳聚糖水溶液各组分配比为:2份低聚壳聚糖,98份水。
对苹果轮纹病菌抑制活性研究中,每皿培养基装量为100mL,微乳剂加入量为2mL,置入的苹果轮纹病菌的菌饼直径为5mm。
对桃褐腐病菌抑制活性研究中,每皿培养基装量为100mL,微乳剂加入量为1mL,置入的桃褐腐病菌的菌饼直径为5mm。
实施例1-4中所制备的复合微乳剂及茶树油乳剂、艾油乳剂和低聚壳聚糖水溶液稀释50倍后对苹果轮纹病菌的抑制率分别达到67.14%,81.12%,62.42%,70.71%,43.38%,50.22%和28.59%;稀释100倍后对桃褐腐病菌的抑制率分别达到63.17%,85.71%,62.38%,83.95%,40.21%,52.39%和22.43%。
(4)对苹果保鲜试验
取表面无病虫害、无损伤的新鲜苹果140个,单果质量为150±10g,直径为6±10Cm。随机分成7组,每组20个苹果。将苹果完全浸入稀释100倍的实施例1-4制备微乳剂中1-2min,捞出自然风干后,放入聚乙烯塑料袋中,不封口。对照组分别采用上述茶树油乳剂、艾油乳剂和低聚壳聚糖水溶液处理,处理方法相同。将处理后苹果在10-15℃、相对湿度为50-80%的环境条件下贮存90天,测定苹果的腐烂率和Vc下降率。结果见表1。
表1 实施例1-4中所制备的复合微乳剂及对照组对苹果的保鲜效果
Vc含量测定方法使用钼蓝法;
(5)对樱桃保鲜试验
取新采摘、表面无损伤樱桃果实700颗,随机分7组,每组100颗。将樱桃完全浸入稀释200倍的实施例制备微乳剂中1-2min,捞出自然风干后,放入聚乙烯塑料袋中,不封口。对照组分别采用上述茶树油乳剂、艾油乳剂和低聚壳聚糖水溶液处理,处理方法相同。将处理后的樱桃在常温(20-25℃)条件下贮藏7d。结果发现,与清水处理对比,实施例制备微乳剂可将樱桃腐烂率降低55%以上。同时还能保持樱桃品质,提高其商品性。实施例1-4制备的微乳剂及对照组对樱桃的保鲜效果见表2。
表2 实施例1-4制备的微乳剂及对照组对樱桃的保鲜效果
Claims (4)
1.一种植物精油-低聚壳聚糖复合抗植物真菌微乳剂,其特征在于:按重量百分比计由以下原料经乳化、复配而成:植物精油5-30%、乳化剂15-30%、助溶剂5-10%、低聚壳聚糖0.5-2%、石榴皮提取物0.5-1%,余量为水;所述植物精油为艾油和茶树油的混合物;所述低聚壳聚糖为相对分子质量低于5kDa的低聚壳聚糖。
2.根据权利要求1所述的植物精油-低聚壳聚糖复合抗植物真菌微乳剂,其特征在于:所述乳化剂为吐温-80、OP-10、Turmul5030、AEO-5中的一种或多种。
3.根据权利要求1所述的植物精油-低聚壳聚糖复合抗植物真菌微乳剂,其特征在于:所述助溶剂为聚乙二醇、聚乙烯醇、丙三醇中的一种。
4.根据权利要求1所述的植物精油-低聚壳聚糖复合抗植物真菌微乳剂,其特征在于:所述石榴皮提取物需要经过微滤膜纯化处理。
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