CN101773238B一种青蒿素的应用----制备方法以及作为杀菌剂和保鲜剂在果蔬保鲜利用和采后病害防治中的应用|
Entry posted by Roger in Artemisinin Patent｜青蒿素专利
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The invention discloses Artemisia apiacea extract and a preparation method and application thereof. The Artemisia apiacea extract is prepared by waste Artemisia apiacea residue discarded during the production of Artemisinin or directly by Artemisia apiacea plant, and has inhibiting effect on pathogenic bacteria of microbe diseases of picked fruits and vegetables and especially has obvious bacteria inhibition in vitro of picked fruit germs such as orange green mould, banana and mango anthracnose, Peronophythora litchi, grape gray mould and the like. Moreover, under living body conditions, the Artemisia apiacea extract can prevent and control diseases of the picked fruits such as oranges, litchi, bananas, grapes and the like, has good effect on the harmless preservation of fruits and vegetables, and can be used as botanical bacteriostat or fruit and vegetable preservative.
本发明公开了一种青蒿提取物和制备方法及应用。所述青蒿提取物是利用生产青蒿素过程废弃的青蒿废渣或者直接由青蒿植株制备得到，其对多种果蔬采后微生物病害的病原菌具有抑制作用，尤其是对柑桔青绿霉菌、香蕉和芒果炭疽病、荔枝霜疫霉病、葡萄灰霉病等水果的采后病菌具有明显的离体抑菌效果，而且在活体条件下，能防治柑桔、荔枝、香蕉、葡萄等水果的采后病害，对果蔬的无害保鲜具有良好的作用，可作为植物源抑菌剂或果蔬保鲜剂使用。
一种青蒿提取物的应用
技术领域
本发明属于果蔬保鲜技术和农药技术领域，具体涉及一种植物提取物的制备和应用，特别是指一种青蒿提取物、其制备方法及其作为杀菌剂和保鲜剂在果蔬保鲜和采后病害防治中的应用。
背景技术
果蔬采后贮藏过程中因微生物病害引起的腐烂所造成的损失是十分巨大的，据资料报道，发达国家的果蔬在采收后因腐烂而造成的损失在15～24％左右，发展中国家由于缺乏冷藏设备及卫生条件较差，果蔬采后病害导致的损失可高达20～50％。
目前国内外应用于采后果蔬保鲜的化学防腐剂主要有施保功、扑海因(抑菌脲)、特克多、多菌灵、甲基托布津、仲丁胺等。然而，长期使用化学杀菌剂极易导致病菌产生抗药性，并导致这些杀菌剂的使用剂量不得不不断加大，从而也引发了食品安全、环境污染和有害生物抗药性等一系列严重问题。人们越来越担心食物链中化学杀菌剂残留对人类健康的影响，要求降低果蔬产品化学杀菌剂的使用量，许多生产上常用的杀菌剂已被许多国家明令禁止在果蔬保鲜中使用。因此，迫切要求研究者寻找新的保鲜药剂用于水果蔬菜的采后保鲜及控制采后果蔬的微生物病害。为了解决上述问题，人们加大了生物农药的开发力度，植物源农药作为生物农药的重要组成部分而倍受重视，国内外均有植物源农药的研发报道。
青蒿为菊科植物黄花蒿(Artemisia annua L.)的全草，别名香蒿、苦蒿、黄蒿。为一年生草本，高达1.5m，全株黄绿色，有臭气。茎直立，具纵条纹， 上部分枝。基部及下部叶在花期枯萎，中部叶卵形，长4.5～7cm，2～3回羽状深裂，小裂片线形，宽约0.3mm，先端尖锐，表面颜色深绿，背面颜色较浅，无毛或略具细微软毛，有柄；上部叶渐小，无柄。头状花序多数，球形，直径约2mm，具细软短梗，排成圆锥状；总苞片2～3层；花管状，雌花长约0.5mm，两性花长约1mm，黄色；花药先端尖尾状，基部圆钝；柱头2裂，裂片先端呈画笔状。瘦果椭圆形，长约0.6mm。花期8～10月，果期10～11月。多生于山坡、林缘、荒地，产于全国各地。青蒿含有抗炎、免疫抑制等生物活性物质，在我国被用于中草药，此外，还可用于提取香料；更重要的是能从青蒿中分离出青蒿素。青蒿素(Artemisinin)是我国药学工作者1971年从青蒿叶中提取分离到的一种具有过氧桥的倍半萜内酯类化合物，是我国自主开发的、国际公认的抗疟疾的特效药，是目前所知的所有抗疟疾药中起效最快，疗效最好，毒性最低的药物，对脑型疟疾和抗氯喹恶性疟疾疗效更为突出，更可贵的是目前青蒿素类药尚未见有关抗药性的报道。
中国是青蒿素的主要产地，占全球总产量的90％。据估计，中国青蒿的种植面积约有7000～8000公顷。考虑到野生青蒿的收获量，估计国内每年可收获青蒿鲜草2万吨左右，青蒿素的理论年产量可达100吨左右，2006年我国青蒿素产量约为78吨。青蒿素提取率约为0.6％左右，因此青蒿原料在提炼青蒿素后会留下大量废渣。以年生产30吨青蒿素计，需要青蒿药材达5000吨以上，留下的废渣以千吨计，均被废弃。
发明内容
本发明的一个目的是利用现有青蒿加工提取过程产生的大量废渣，提供一种从青蒿废渣中提取的提取物，变废为宝。
本发明的另一个目的是提供所述青蒿提取物的提取方法。
本发明还有一个目的是提供所述青蒿提取物的新应用，所述青蒿提取物也包括直接采用青蒿进行提取得到的提取物，而不仅仅包括采用青蒿废渣制备得到的提取物。
本发明的目的通过以下技术方案予以实现：
提供一种青蒿提取物，方案一是以青蒿废渣浸膏为原料，往浸膏中加入溶剂，浸提，过滤得到的滤液浓缩或干燥得到所述青蒿提取物。
所述青蒿废渣浸膏是将青蒿全株或切段后晒干、阴干或烘干，粉碎后采用体积比浓度为60～70％的乙醇浸提过夜，浸提液用于提取青蒿素，剩余部分即为废渣，浸提液在精提青蒿素过程中也得到部分废渣，将收集的各阶段产生的废渣浓缩成废渣浸膏即可，也可以混合各阶段收集到的废渣统一浓缩成浸膏，浓缩成浸膏的方法为现有常规方法；或者从青蒿素加工工厂直接购买被废弃的青蒿废渣浸膏。
所述溶剂可采用本领域常用的溶剂，优选水、乙醇、乙酸乙酯、石油醚或丙酮中任何一种或几种溶剂任意比例的混合物。
本发明所述青蒿提取物的提取方法具体包括以下步骤：
(1)往青蒿废渣浸膏中加入硅胶，搅拌均匀，磨粉过筛得青蒿废渣浸膏粉；
(2)往步骤(1)所得青蒿废渣浸膏粉中加入溶剂浸提后过滤；
(3)将步骤(2)所得滤液经旋转浓缩或干燥得青蒿废渣提取物。
作为优选，步骤(3)得到的滤液经旋转浓缩后得青蒿废渣提取液，提取液再用溶剂进行液-液或液-固分部提取，提取液及其分部提取液经浓缩或干燥 得到青蒿废渣提纯后的提取物。液-液或液-固分部提取所采用的溶剂优选水、乙醇、乙酸乙酯、石油醚或丙酮中任何一种或几种溶剂任意比例的混合物，溶剂的加入量为(重量体积比)为1g∶0.5～2ml；提取24小时。
步骤(1)所述硅胶的加入量按照1000g青蒿废渣浸膏加入500～2000g硅胶的比例确定；所述过筛为过30～100目筛。
步骤(2)所述溶剂为水、乙醇、乙酸乙酯、石油醚或丙酮中任何一种或几种溶剂任意比例的混合物，加入的量按照废渣浸膏粉∶溶剂的重量体积比为1g∶0.5～2ml的比例确定。
步骤(2)所述浸提的时间为5～24小时，优选24小时。
步骤(3)所述旋转浓缩或干燥均为常规方法。
方案二，提供一种青蒿提取物，将青蒿全株或切段后晒干(阴干或烘干)粉碎后采用水或其他常规溶剂浸提得到。
本发明方案一和方案二所述青蒿提取物都具有新的应用价值，可以应用作为杀菌剂或果蔬保鲜剂。所述青蒿提取物包括采用青蒿废渣或青蒿植株提取得到的提取液、浓缩提取液或干燥得到的固体提取物。青蒿提取物的抗菌保鲜应用是由本发明首次提出的技术方案，并不仅仅限于青蒿废渣的再利用。青蒿整株植物也可提取抗菌保鲜物质，因为保鲜剂的经济价值远远低于青蒿素，青蒿主要用于提取青蒿素，不会仅用于提取保鲜剂。所以本发明不仅提供了青蒿提取物的新应用，更重要的是解决了青蒿素生产过程中产生的大量废渣的再利用。
本发明经过反复试验证明，从青蒿或青蒿废渣中提取得到的提取物对多种果蔬采后微生物病害的病原菌具有抑制作用，尤其是对柑桔青绿霉菌、香蕉和 芒果炭疽病、荔枝霜疫霉病、葡萄灰霉病等水果的采后病菌具有明显的离体抑菌效果，而且在活体条件下，能防治柑桔、荔枝、香蕉等水果的采后病害，对果蔬的无害保鲜具有良好的作用。因此，从青蒿废渣中得到的提取物可作为植物源抑菌剂或果蔬保鲜剂使用。
本发明的有益效果是：
本发明提供了一种新的植物源杀菌剂或果蔬保鲜剂，为抗菌和蔬果保鲜技术领域提供新的技术成果。
本发明采用适当的方法处理废弃的青蒿废渣后再进行溶剂提取制备得到有益的抗菌保鲜剂，变废为宝；同时解决了长期以来青蒿素加工企业处置废渣的难题，为推广青蒿素的生产加工应用扫平障碍。
本发明方法简单易行，原料易得且成本较低，具有优良的推广价值。
具体实施方式
下面结合具体实施例进一步详细说明本发明。
实施例1 青蒿提取物的制备
青蒿废渣浸膏来自广东省某青蒿素生产厂家，其他试剂均为市购常规产品。1000g青蒿浸膏加入1000g硅胶，搅拌均匀，磨成粉状，过60目筛，快速分离，得青蒿废渣浸膏粉；用1800mL石油醚(PE)浸提青蒿废渣浸膏粉24小时，过滤或离心得到PE部分的滤液732mL，经旋转浓缩后得PE部分提取物157.65g，所得青蒿浸膏提取物低温(1～7℃)贮存备用。
实施例2 青蒿提取物的制备
青蒿废渣浸膏来自广东省某青蒿素生产厂家，其他试剂均为市购常规产品。1000g青蒿浸膏加入1000g硅胶，搅拌均匀，磨成粉状，过60目筛。快 速分离得青蒿废渣浸膏粉；用1800mL的乙酸乙酯(EA)浸提青蒿废渣浸膏粉24小时，过滤或离心得到EA部分的滤液1468mL，经旋转浓缩后得EA部分提取物143.58g，低温贮存备用。
实施例3 青蒿提取物的制备
青蒿废渣浸膏来自广东省某青蒿素生产厂家，其他试剂均为市购常规产品。1000g青蒿浸膏加入1000g硅胶，搅拌均匀，磨成粉状，过60目筛。快速分离得青蒿废渣浸膏粉；用1800mL的PE∶EA＝1∶1(体积比)的混合液浸提青蒿废渣浸膏粉24小时，过滤或离心得到混合液部分的滤液1392mL，经旋转浓缩后得混合液部分提取物223.4g，低温贮存备用。
实施例4 青蒿提取物的制备
青蒿废渣浸膏来自广东省某青蒿素生产厂家，其他试剂均为市购常规产品。1000g青蒿浸膏加入2000g硅胶，搅拌均匀，磨成粉状，过30目筛。快速分离得青蒿废渣浸膏粉；用1800mL的PE∶EA＝1∶1(体积比)的混合液浸提青蒿废渣浸膏粉24小时，过滤或离心得到混合液部分的滤液1314mL，经旋转浓缩后得混合液部分提取物201.9g，低温贮存备用。
实施例5 青蒿提取物的制备
青蒿废渣浸膏来自广东省某青蒿素生产厂家，其他试剂均为市购常规产品。1000g青蒿浸膏加入1500g硅胶，搅拌均匀，磨成粉状，过80目筛，快速分离，得青蒿废渣浸膏粉；用1800mL石油醚(PE)浸提青蒿废渣浸膏粉24小时，过滤或离心得到PE部分的滤液685mL，经旋转浓缩后得PE部分提取物135.8g，所得青蒿浸膏提取物低温(1～7℃)贮存备用。
实施例6 青蒿提取物的制备
将青蒿全株或切段的青蒿晒干(阴干或烘干)，粉碎，过40～60目筛。按粉碎物∶水＝1∶5(重量/体积，g/ml)的比例，提取24小时，过滤或离心得到提取液，再经常规真空浓缩得浓缩液或干燥的固体提取物，低温贮存备用。
实施例7 青蒿提取物的制备
将青蒿全株或切段的青蒿晒干(阴干或烘干)，粉碎，过40～60目筛。按粉碎物∶60％的乙醇＝1∶1(重量/体积，g/ml)的比例，提取24小时，过滤或离心得到提取液，再经常规真空浓缩得浓缩液或干燥的固体提取物，低温贮存备用。
实施例8 青蒿废渣不同溶剂或方法提取物对水果采后病原菌的抑制作用试验
采用改进的生长速率法，称取本发明不同溶剂的青蒿提取物与NYDA培养基混合均匀，制成浓度为500μg青蒿提取物/mL培养基、1000μg青蒿提取物/mL培养基和2000μg青蒿提取物/mL培养基的带药培养基，将带药培养基倒入直径为9cm的培养皿中，冷却后，在培养皿中央接种5mm沙糖桔指状青霉菌(Penicillium digitatum)或香蕉炭疽病菌(Collectotrichun musae)菌片(所述菌种来自华南农业大学植物病理系实验室，也可以采用常规方法分离得到)，以纯NYDA培养基平板作为空白对照。置于28℃恒温箱中培养，测量记录菌落大小(用十字交叉法测定菌落直径)，计算抑菌率。
抑菌率(％)＝(对照菌落直径-处理菌落直径)/对照菌落直径×100
实验结果见表1和表2。从表1中可以发现，本发明青蒿不同溶剂提取物对沙糖桔指状青霉的生长均有一定的抑制作用，以乙酸乙酯提取物的抑菌效果最强。从表2中可以发现，与抑制沙糖桔指状青霉菌效果相比，本发明青蒿提取物对香蕉炭疽病菌的抑菌效果更好，也以乙酸乙酯提取物的抑菌效 果最强。
表1 青蒿废渣不同溶剂提取物对采后沙糖桔指状青霉病菌抑菌效果
表2 青蒿废渣不同溶剂提取物对采后香蕉炭疽病菌抑菌效果
实施例9 本发明青蒿提取物对沙糖桔采后绿霉病的抑制效果试验
将本发明青蒿提取物加水配成1000μg/mL溶液，沙糖桔在所述溶液中浸泡5min后晾干，用灭菌铁钉在果实近果蒂部区域对称打2个孔(深3mm，直径2mm)，10μl 1.5×104CFU/mL指状青霉孢子菌悬液。待自然晾干后放到塑料筐内于20℃，85％～95％RH条件下贮藏22天，统计病情指数和防治效果。试验设空白对照，并以水果保鲜方面常用的施保功(Sporgon)作为商品杀菌剂对照。实验结果见表3。从表3可以看出，本发明青蒿提取物对沙糖桔绿霉病(由指状青霉菌引起)均有明显效果，但比商品杀菌剂施保功效果略差。
表3 本发明青蒿提取物对沙糖桔指状青霉的活体抑制效果
(20℃、90％RH下培养22天)
实施例10 本发明青蒿提取物对荔枝采后病害的控制效果试验
荔枝采后病害主要是霜疫霉病。霜疫霉病是目前荔枝果实上最严重的病害，在荔枝采后贮运期间造成严重损失，使荔枝保鲜与销售企业蒙受重大损失。目前主要依靠化学杀菌剂如瑞毒霉、乙磷铝、特克多、施保功等控制。将本发明青蒿废渣的乙酸乙酯提取物加水配成1000μg/mL溶液，采收后的荔枝果实(品种：淮枝)在所述溶液中浸泡5min，待自然晾干后放到塑料筐内于20℃，85％～95％RH条件下贮藏7天，统计病情指数和防治效果。试验设空白对照，并以荔枝果实保鲜最常用的施保功(Sporgon)作为商品杀菌剂对照。试验结果见表4。从表4可以看出，本发明青蒿废渣的乙酸乙酯提取物对荔枝果实采后病害(主要由由霜疫霉菌引起)有明显效果，但比商品杀菌剂施保功效果略差。
表4 本发明青蒿提取物对荔枝采后病害的控制效果
(20℃、90％RH下培养22天)
实施例11 本发明青蒿提取物对葡萄灰霉病抑制效果试验
葡萄采后病害主要是灰霉病，90％以上的葡萄果实腐烂是由于该病的爆发引起在葡萄采后贮运期间造成严重损失。目前主要依靠二氧化硫熏蒸或二氧化硫释放纸进行控制，但二氧化硫处理容易导致葡萄表面漂白，特别是果蒂部分漂白严重；另外，也容易导致二氧化硫残留超标。将本发明青蒿废渣的乙酸乙酯提取物加水配成1000μg/mL溶液，采收后的葡萄果实(品种：巨峰)在所述溶液中浸泡5min，待自然晾干后放到塑料筐内于20℃，85％～95％RH条件下贮藏20天，统计病情指数和防治效果。试验设空白对照。试验结果见表5。从表5可以看出，本发明青蒿废渣的乙酸乙酯提取物对巨峰葡萄果实采后病害(主要由由灰霉菌引起)有明显效果。
表4 本发明青蒿提取物对葡萄采后病害的控制效果
(20℃、90％RH下培养20天)
1.一种青蒿提取物的应用，其特征在于应用于制备防治柑桔青绿霉菌、香蕉炭疽病或荔枝霜疫霉病的制剂，所述青蒿提取物是采用青蒿废渣浸膏为原料，加入溶剂浸提，过滤得到的滤液再经浓缩或干燥得到，所述溶剂为水、乙醇、乙酸乙酯、石油醚或丙酮中任何一种或几种溶剂任意比例的混合物；所述青蒿废渣浸膏是将青蒿全株或切段后晒干、阴干或烘干，粉碎后采用体积比浓度为60-70％的乙醇浸提过夜，浸提液用于提取青蒿素，剩余部分即为废渣，浸提液在精提青蒿素过程中也得到废渣，将收集的各阶段产生的废渣浓缩得到废渣浸膏。
2.根据权利要求1所述青蒿提取物的应用，其特征在于所述青蒿提取物的制备方法包括以下步骤：
(1)向青蒿废渣浸膏中加入硅胶，搅拌均匀，磨粉过筛得青蒿废渣浸膏粉；
(2)向步骤(1)所得青蒿废渣浸膏粉中加入溶剂浸提后过滤得滤液，所述溶剂为水、乙醇、乙酸乙酯、石油醚或丙酮中任何一种或几种溶剂任意比例的混合物；
(3)将步骤(2)所得滤液经旋转浓缩或干燥得青蒿废渣提取物。
3.根据权利要求2所述青蒿提取物的应用，其特征在于步骤(1)所述硅胶的加入量按照1000g青蒿废渣浸膏加入500～2000g硅胶的比例确定；所述过筛为过30～100目筛。
4.根据权利要求2所述青蒿提取物的应用，其特征在于步骤(2)所述溶剂加入的量按照废渣浸膏粉：溶剂的重量体积比为1g∶0.5～2ml的比例确定。
5.根据权利要求2所述青蒿提取物的应用，其特征在于步骤(2)所述浸提的时间为24小时。
The invention discloses Artemisia apiacea extract and a preparation method and application thereof. The Artemisia apiacea extract is prepared by waste Artemisia apiacea residue discarded during the production of Artemisinin or directly by Artemisia apiacea plant, and has inhibiting effect on pathogenic bacteria of microbe diseases of picked fruits and vegetables and especially has obvious bacteria inhibition in vitro of picked fruit germs such as orange green mould, banana and mango anthracnose, Peronophythora litchi, grape gray mould and the like. Moreover, under living body conditions, the Artemisia apiacea extract can prevent and control diseases of the picked fruits such as oranges, litchi, bananas, grapes and the like, has good effect on the harmless preservation of fruits and vegetables, and can be used as botanical bacteriostat or fruit and vegetable preservative.
A kind of application of Artemisinin
Technical field
The invention belongs to postharvest technology of fruits and vegetables and technical field of pesticide; Be specifically related to a kind of preparation and application of plant extracts, be meant a kind of Artemisinin, its preparation method especially and as bactericide and the application of antistaling agent in preserving fruit and vegetable utilizing and postharvest disease control.
Background technology
The loss of rotting to be caused that causes because of microorganism disease in the postharvest fruit and vegetable storage is very huge; According to document announcement; The fruits and vegetables of developed country are lost in about 15～24% because of what rot to cause in the back of gathering; Developing country is because shortage refrigerating equipment and sanitary condition are relatively poor, and the loss that postharvest disease of fruits and vegetables causes can be up to 20～50%.
Domestic and international application mainly contains sporgon, iprodione (antibacterial urea), Te Keduo, carbendazim, thiophanate methyl, sec-butylamine etc. in the chemical preservative of adopting the back preserving fruit and vegetable utilizing at present.Yet, use chemical bactericide very easily to cause germ to develop immunity to drugs for a long time, and cause the using dosage of these bactericide to have to continue to increase, thereby also caused food security, a series of serious problems such as environmental pollution and harmful organism resistance to the action of a drug.People more and more is worried the residual influence to human health of chemical bactericide in the food chain, requires to reduce the use amount of fruit and vegetable food chemical bactericide, and bactericide commonly used has been prohibited in preserving fruit and vegetable utilizing by many countries and used in many productions.Therefore, an urgent demand researcher seeks the microorganism disease that the back fruits and vegetables are adopted in post-harvest fresh-keeping and control that new storage chemical is used for fruit and vegetable.In order to address the above problem, people have strengthened the exploitation dynamics of biological pesticide, and botanical pesticide is extremely paid attention to as the important component part of biological pesticide, and the research and development report of botanical pesticide is all arranged both at home and abroad.
Sweet wormwood is the herb of feverfew artemisia annua (Artemisia annua L.), another name abrotanum, Artemisia santonica, yellow wormwood artemisia.Be annual herb, up to 1.5m, the complete stool yellow green has foul smell.Stem is upright, tool vertical stripe, top branch.Base portion and bottom leaf are withered at the florescence, and middle leaf is avette, long 4.5～7cm, and 2～3 times pinniform drastic cracks, lobelet is linear, wide about 0.3mm, tip is sharp-pointed, and surface color is dark green, and back side lighter color does not have hair or omits the trickle fur of tool, and handle is arranged; Upper leaf is gradually little, stockless.Capitulum is most, sphere, and the about 2mm of diameter, the soft short stalk of tool is lined up coniform; 2～3 layers of phyllarys; The floral tube shape, female flower is about 0.5mm, and hermaphrodite flower is about 1mm, yellow; Flower pesticide tip fine stern shape, the base portion circle is blunt; Column cap 2 splits, and the sliver tip is penicillate.Achene is oval, is about 0.6mm.8～October of florescence, really 10～November of phase.Be born in hillside, border, wasteland more, originate in all parts of the country.Sweet wormwood contains bioactivators such as anti-inflammatory, immunosupress, is used to Chinese herbal medicine in China, in addition, also can be used for extracting spices; The more important thing is and from sweet wormwood, to isolate qinghaosu.Qinghaosu (Artemisinin) is that China pharmacy worker extracted a kind of sesquiterpene lactone compounds with peroxide bridge that is separated in 1971 from artemisia leaf; Be China's independent development, internationally recognized antimalarial specific drug; Be that onset is the fastest in all Anti-Malarials known today, curative effect is best, the medicine that toxicity is minimum; More outstanding to encephalic malaria and anti-chloroquine pernicious malaria curative effect, more valuable is that present arteannuin medicine is not seen relevant drug-fast report as yet.
China is the main place of production of qinghaosu, accounts for 90% of global total output.According to estimates, the cultivated area of Chinese wormwood has 7000～8000 hectares approximately.Consider the harvest yield of wild sweet wormwood, estimate can gather in the crops domestic every year about 20,000 tons of the bright grass of sweet wormwood, the theoretical annual production of qinghaosu can reach about 100 tons, and China's qinghaosu output was about 78 tons in 2006.The qinghaosu recovery rate is about about 0.6%, so the sweet wormwood raw material can stay a large amount of waste residues after refining qinghaosu.Year to produce 30 tons of qinghaosus, need the sweet wormwood medicinal material to reach more than 5000 tons, the waste residue that stays all goes out of use in kiloton.
Summary of the invention
An object of the present invention is to utilize a large amount of waste residues of existing sweet wormwood processing leaching process generation, a kind of extract that from the sweet wormwood waste residue, extracts is provided, turn waste into wealth.
Another object of the present invention provides the method for distilling of said Artemisinin.
A further object of the invention provides the new application of said Artemisinin, and said Artemisinin comprises that also direct employing sweet wormwood extracts the extract that obtains, and not only comprises the extract that adopts the sweet wormwood waste residue to prepare.
The object of the invention is achieved through following technical scheme:
A kind of Artemisinin is provided, and scheme one is to be raw material with sweet wormwood waste residue medicinal extract, in medicinal extract, adds solvent, and lixiviate filters that the filtrating that obtains concentrates or drying obtains said Artemisinin.
Said sweet wormwood waste residue medicinal extract is with drying, dry in the shade or dry after sweet wormwood complete stool or the segment; Pulverizing the back, to adopt volume by volume concentration be that 60～70% ethanol lixiviate is spent the night, and leaching liquor is used to extract qinghaosu, and remainder is waste residue; Leaching liquor is carried in essence and is also obtained the part waste residue in the qinghaosu process; The waste residue that each stage of collecting is produced is condensed into waste residue medicinal extract and get final product, also can mix the waste residue unification that each stage collects and be condensed into medicinal extract, and the method that is condensed into medicinal extract is for having conventional method now; Perhaps directly buy the sweet wormwood waste residue medicinal extract that goes out of use from qinghaosu processing factory.
Said solvent can adopt this area solvent commonly used, the mixture of any or several kinds of solvent arbitrary proportions in preferred water, ethanol, ethyl acetate, benzinum or the acetone.
The method for distilling of Artemisinin according to the invention specifically may further comprise the steps:
(1) in sweet wormwood waste residue medicinal extract, add silica gel, stir, abrasive dust sieve sweet wormwood waste residue extract powder;
(2) filter after adding the solvent lixiviate in step (1) the gained sweet wormwood waste residue extract powder;
(3) step (2) gained filtrating is concentrated or the dry sweet wormwood waste residue extract that gets through rotation.
As preferably, the filtrating that step (3) obtains through rotate after concentrating sweet wormwood waste residue extract, extract carries out liquid-liquid with solvent again or liquid-solid branch extracts, extract and branch's extract thereof are through concentrating or dry extract after obtaining the purification of sweet wormwood waste residue.Liquid-liquid or liquid-solid branch extract the mixture of any or several kinds of solvent arbitrary proportions in solvent preferred water, ethanol, ethyl acetate, benzinum or the acetone that is adopted, and the addition of solvent be that (w/v) is 1g: 0.5～2ml; Extracted 24 hours.
The addition of the said silica gel of step (1) is confirmed according to the ratio that 1000g sweet wormwood waste residue medicinal extract adds 500～2000g silica gel; Said sieving to crossing 30～100 mesh sieves.
The said solvent of step (2) is the mixture of any or several kinds of solvent arbitrary proportions in water, ethanol, ethyl acetate, benzinum or the acetone, and the amount of adding is according to the waste residue extract powder: the w/v of solvent is that the ratio of 1g: 0.5～2ml is confirmed.
The time of the said lixiviate of step (2) is 5～24 hours, preferred 24 hours.
The said rotation of step (3) concentrates or drying is conventional method.
Scheme two provides a kind of Artemisinin, pulverizes the back and adopts water or other conventional solvent lixiviates to obtain drying (dry in the shade or dry) after sweet wormwood complete stool or the segment.
The present invention program one has new using value with scheme two said Artemisinins, can use as bactericide or fruit and vegetable fresh-keeping agent.Said Artemisinin comprises extract, concentrated extracting solution or the dry solid extract that obtains that adopts sweet wormwood waste residue or the extraction of sweet wormwood plant to obtain.It is the technical scheme that is proposed first by the present invention that the anti-bacterial refreshing of Artemisinin is used, and is not restricted to the utilization again of sweet wormwood waste residue.The whole strain plant of sweet wormwood also can be extracted the anti-bacterial refreshing material, because the economic worth of antistaling agent is well below qinghaosu, sweet wormwood is mainly used in the extraction qinghaosu, can only not be used to extract antistaling agent.So the present invention not only provides the new application of Artemisinin, the more important thing is the utilization again that has solved a large amount of waste residues that produce in the qinghaosu production process.
The present invention proves through repetition test; It is inhibited to the pathogen of multiple postharvest fruit and vegetable microorganism disease from sweet wormwood or sweet wormwood waste residue, to extract the extract that obtains; Especially the back germ of adopting to fruit such as citrus penicillium, banana and mango anthracnose, peronophythora litchi, grape grey moulds has the fungistatic effect that significantly exsomatizes; And under condition of living body, can prevent and treat the postharvest disease of fruit such as citrus, lichee, banana, fruits and vegetables harmless fresh-keeping had good effect.Therefore, the extract that from the sweet wormwood waste residue, obtains can be used as plant-based bacteriostat or fruit and vegetable fresh-keeping agent uses.
The invention has the beneficial effects as follows:
The invention provides a kind of new botanical fungicide or fruit and vegetable fresh-keeping agent, with vegetables and fruits preservation technique field new technological achievement is provided for antibiotic.
Carry out solvent extraction again behind the sweet wormwood waste residue that the present invention adopts the appropriate method processing to discard and prepare useful antimicrobial antistaling agent, turn waste into wealth; Solved the difficult problem of the waste residue of qinghaosu processing enterprise disposal for a long time simultaneously, used for the production and processing of promoting qinghaosu and put down obstacle.
The inventive method is simple, raw material be easy to get and cost lower, have good promotional value.
The specific embodiment
Below in conjunction with specific embodiment further explain the present invention.
The preparation of embodiment 1 Artemisinin
Sweet wormwood waste residue medicinal extract is from Guangdong Province's qinghaosu manufacturer, and other reagent are commercial conventional products.1000g sweet wormwood medicinal extract adds 1000g silica gel, stirs, and pulverize is crossed 60 mesh sieves, separates fast, gets sweet wormwood waste residue extract powder; With 1800mL benzinum (PE) lixiviate sweet wormwood waste residue extract powder 24 hours, filter or the centrifugal filtrating 732mL that obtains the PE part, after rotation concentrates PE extracting section thing 157.65g, gained sweet wormwood medicinal extract extract low temperature (1～7 ℃) storage is subsequent use.
The preparation of embodiment 2 Artemisinins
Sweet wormwood waste residue medicinal extract is from Guangdong Province's qinghaosu manufacturer, and other reagent are commercial conventional products.1000g sweet wormwood medicinal extract adds 1000g silica gel, stirs, and pulverize is crossed 60 mesh sieves.Separate fast sweet wormwood waste residue extract powder; With ethyl acetate (EA) the lixiviate sweet wormwood waste residue extract powder of 1800mL 24 hours, filter or the centrifugal filtrating 1468mL that obtains the EA part,, rotation gets EA extracting section thing 143.58g after concentrating, and low tempertaure storage is subsequent use.
The preparation of embodiment 3 Artemisinins
Sweet wormwood waste residue medicinal extract is from Guangdong Province's qinghaosu manufacturer, and other reagent are commercial conventional products.1000g sweet wormwood medicinal extract adds 1000g silica gel, stirs, and pulverize is crossed 60 mesh sieves.Separate fast sweet wormwood waste residue extract powder; PE with 1800mL: EA=1: the mixed liquor lixiviate sweet wormwood waste residue extract powder of 1 (volume ratio) 24 hours, filter or the centrifugal filtrating 1392mL that obtains the mixed liquor part,, rotation gets mixed liquor extracting section thing 223.4g after concentrating, and low tempertaure storage is subsequent use.
The preparation of embodiment 4 Artemisinins
Sweet wormwood waste residue medicinal extract is from Guangdong Province's qinghaosu manufacturer, and other reagent are commercial conventional products.1000g sweet wormwood medicinal extract adds 2000g silica gel, stirs, and pulverize is crossed 30 mesh sieves.Separate fast sweet wormwood waste residue extract powder; PE with 1800mL: EA=1: the mixed liquor lixiviate sweet wormwood waste residue extract powder of 1 (volume ratio) 24 hours, filter or the centrifugal filtrating 1314mL that obtains the mixed liquor part,, rotation gets mixed liquor extracting section thing 201.9g after concentrating, and low tempertaure storage is subsequent use.
The preparation of embodiment 5 Artemisinins
Sweet wormwood waste residue medicinal extract is from Guangdong Province's qinghaosu manufacturer, and other reagent are commercial conventional products.1000g sweet wormwood medicinal extract adds 1500g silica gel, stirs, and pulverize is crossed 80 mesh sieves, separates fast, gets sweet wormwood waste residue extract powder; With 1800mL benzinum (PE) lixiviate sweet wormwood waste residue extract powder 24 hours, filter or the centrifugal filtrating 685mL that obtains the PE part, after rotation concentrates PE extracting section thing 135.8g, gained sweet wormwood medicinal extract extract low temperature (1～7 ℃) storage is subsequent use.
The preparation of embodiment 6 Artemisinins
The sweet wormwood of sweet wormwood complete stool or segment is dried (dry in the shade or dry), pulverize, cross 40～60 mesh sieves.Press crushed material: water=1: 5 (weight per volume, ratio g/ml) were extracted 24 hours, filter or the centrifugal extract that obtains, again through conventional vacuum concentrate concentrate or dry solid extract, low tempertaure storage is subsequent use.
The preparation of embodiment 7 Artemisinins
The sweet wormwood of sweet wormwood complete stool or segment is dried (dry in the shade or dry), pulverize, cross 40～60 mesh sieves.Press crushed material: 60% ethanol=1: 1 (weight per volume, ratio g/ml) were extracted 24 hours, filter or the centrifugal extract that obtains, again through conventional vacuum concentrate concentrate or dry solid extract, low tempertaure storage is subsequent use.
Embodiment 8 sweet wormwood waste residue different solvents or method extract are adopted the inhibitory action test of back pathogen to fruit
Adopt improved growth rate method; The Artemisinin and the NYDA culture medium that take by weighing different solvents of the present invention mix; Processing concentration is the band medicine culture medium of 500 μ g Artemisinin/mL culture mediums, 1000 μ g Artemisinin/mL culture mediums and 2000 μ g Artemisinin/mL culture mediums; Band medicine culture medium is poured in the culture dish that diameter is 9cm; After the cooling; At culture dish central authorities inoculation 5mm granulated sugar tangerine penicillium digitatum (Penicillium digitatum) or Glorosprium musarum Cookeet Mass (Collectotrichun musae) bacterium sheet (said bacterial classification also can adopt conventional method to separate and obtain from department of plant pathology of Agricultural University Of South China laboratory), with pure NYDA culture medium flat plate as blank.Place 28 ℃ of insulating boxs to cultivate, surveying record bacterium colony size (measuring colony diameter with the right-angled intersection method) is calculated bacteriostasis rate.
Bacteriostasis rate (%)=(contrast colony diameter-processing colony diameter)/contrast colony diameter * 100
Experimental result is seen table 1 and table 2.Can find that from table 1 sweet wormwood different solvents extract of the present invention all has certain inhibitory action to the growth of granulated sugar tangerine Penicillium digitatum, and is the strongest with the fungistatic effect of ethyl acetate extract.Can find that from table 2 with inhibition granulated sugar tangerine penicillium digitatum comparison, Artemisinin of the present invention is better to the fungistatic effect of Glorosprium musarum Cookeet Mass, also the fungistatic effect with ethyl acetate extract is the strongest.
Table 1 sweet wormwood waste residue different solvents extract is to adopting back granulated sugar tangerine Penicillium digitatum germ fungistatic effect
Table 2 sweet wormwood waste residue different solvents extract is to adopting back Glorosprium musarum Cookeet Mass fungistatic effect
Embodiment 9 Artemisinins of the present invention are adopted the inhibition effect test of back green mould to the granulated sugar tangerine
Artemisinin of the present invention is added water be made into 1000 μ g/mL solution; The granulated sugar tangerine is dried after in said solution, soaking 5min; Make a call to 2 holes (dark 3mm, diameter 2mm), 10 μ l, 1.5 * 104CFU/mL Penicillium digitatum pityrosporion ovale suspension with the sterilization iron nail in the nearly base of fruit of fruit portion zone symmetry.It is interior in 20 ℃ to treat to be put into after nature dries plastic crate, and storage is 22 days under 85%～95%RH condition, statistics disease index and prevention effect.Blank is established in test, and contrasts as the commodity bactericide with fruit freshness preserving aspect sporgon (Sporgon) commonly used.Experimental result is seen table 3.Can find out that from table 3 Artemisinin of the present invention all has positive effect to granulated sugar tangerine green mould (being caused by penicillium digitatum), but slightly poorer than commodity bactericide sporgon effect.
Table 3 Artemisinin of the present invention suppresses effect to the live body of granulated sugar tangerine Penicillium digitatum
(20 ℃, 90%RH were cultivated 22 days down)
Embodiment 10 Artemisinins of the present invention are to the control effect test of lichee postharvest disease
The lichee postharvest disease mainly is a peronospora tabacina.Peronospora tabacina is severe diseases on the present litchi fruits, during lichee is adopted the back storing, causes heavy losses, and lichee fresh-keeping and sale enterprise are sustained a great loss.Controls such as main at present dependence chemical bactericide such as metalaxyl, aliette, Te Keduo, sporgon.The ethyl acetate extract of sweet wormwood waste residue of the present invention is added water be made into 1000 μ g/mL solution; Litchi fruits after gathering (kind: the Huaihe River branch) in said solution, soak 5min; Treat to be put into after nature dries in the plastic crate in 20 ℃; Storage is 7 days under 85%～95%RH condition, statistics disease index and prevention effect.Blank is established in test, and contrasts as the commodity bactericide with the fresh-keeping the most frequently used sporgon of litchi fruits (Sporgon).Result of the test is seen table 4.Can find out that from table 4 ethyl acetate extract of sweet wormwood waste residue of the present invention has positive effect to litchi fruits postharvest disease (mainly by being caused by white phytophthora), but slightly poorer than commodity bactericide sporgon effect.
Table 4 Artemisinin of the present invention is to the control effect of lichee postharvest disease
(20 ℃, 90%RH were cultivated 22 days down)
Embodiment 11 Artemisinins of the present invention suppress effect test to grape grey mould
The grape postharvest disease mainly is a gray mold, and it is to cause heavy losses during the storing because the outburst of this disease causes after the grape harvest that the grape fruit more than 90% rots.Main at present dependence sulfur dioxide is fumigated or sulfur dioxide release paper is controlled, but sulfur dioxide treatment causes the grape superficial bleaching easily, and particularly the base of fruit partial bleaching is serious; In addition, also cause sulfur dioxide residue to exceed standard easily.The ethyl acetate extract of sweet wormwood waste residue of the present invention is added water be made into 1000 μ g/mL solution; Grape fruit after gathering (kind: huge peak) in said solution, soak 5min; Treat to be put into after nature dries in the plastic crate in 20 ℃; Storage is 20 days under 85%～95%RH condition, statistics disease index and prevention effect.Blank is established in test.Result of the test is seen table 5.Can find out that from table 5 ethyl acetate extract of sweet wormwood waste residue of the present invention has positive effect to kyoto grape postharvest diseases of fruit (mainly by being caused by botrytis cinerea).
Table 4 Artemisinin of the present invention is to the control effect of grape postharvest disease
(20 ℃, 90%RH were cultivated 20 days down)
1. the application of an Artemisinin; It is characterized in that being applied to prepare the preparation of control citrus penicillium, banana anthracnose or peronophythora litchi; Said Artemisinin is that employing sweet wormwood waste residue medicinal extract is raw material; Add the solvent lixiviate, filter the filtrating that obtains again through concentrating or drying obtains, said solvent is the mixture of any or several kinds of solvent arbitrary proportions in water, ethanol, ethyl acetate, benzinum or the acetone; Said sweet wormwood waste residue medicinal extract is with drying, dry in the shade or dry after sweet wormwood complete stool or the segment; Employing volume by volume concentration in pulverizing back is that the ethanol lixiviate of 60-70% is spent the night; Leaching liquor is used to extract qinghaosu; Remainder is waste residue, and leaching liquor is carried in essence and also obtained waste residue in the qinghaosu process, with the concentrated waste residue medicinal extract that obtains of the waste residue of each stage generation of collecting.
2. according to the application of the said Artemisinin of claim 1, it is characterized in that the preparation method of said Artemisinin may further comprise the steps:
(1) in sweet wormwood waste residue medicinal extract, add silica gel, stir, abrasive dust sieve sweet wormwood waste residue extract powder;
(2) in step (1) gained sweet wormwood waste residue extract powder, add to cross after the solvent lixiviate and filter filtrating, said solvent is the mixture of any or several kinds of solvent arbitrary proportions in water, ethanol, ethyl acetate, benzinum or the acetone;
(3) step (2) gained filtrating is concentrated or the dry sweet wormwood waste residue extract that gets through rotation.
3. according to the application of the said Artemisinin of claim 2, it is characterized in that the addition of the said silica gel of step (1) is confirmed according to the ratio that 1000g sweet wormwood waste residue medicinal extract adds 500～2000g silica gel; Said sieving to crossing 30～100 mesh sieves.
4. according to the application of the said Artemisinin of claim 2, it is characterized in that amount that the said solvent of step (2) adds is according to the waste residue extract powder: the w/v of solvent is that the ratio of 1g: 0.5～2ml is confirmed.
5. according to the application of the said Artemisinin of claim 2, it is characterized in that the time of the said lixiviate of step (2) is 24 hours.
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