CN105767167A - 一种复合植物提取物及其制备方法与在抑制赭曲霉生长中的应用 - Google Patents
一种复合植物提取物及其制备方法与在抑制赭曲霉生长中的应用 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B9/00—Preservation of edible seeds, e.g. cereals
- A23B9/16—Preserving with chemicals
- A23B9/24—Preserving with chemicals in the form of liquids or solids
- A23B9/26—Organic compounds; Microorganisms; Enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/42—Preservation of non-alcoholic beverages
- A23L2/44—Preservation of non-alcoholic beverages by adding preservatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3481—Organic compounds containing oxygen
- A23L3/349—Organic compounds containing oxygen with singly-bound oxygen
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3481—Organic compounds containing oxygen
- A23L3/3499—Organic compounds containing oxygen with doubly-bound oxygen
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
- C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
本发明公开了一种复合植物提取物及其制备方法与在抑制赭曲霉生长中的应用。一种复合植物提取物,它由肉桂醛、柠檬醛和丁香酚组成。本发明复合植物提取物应用在抑制赭曲霉生长中。本发明复合植物提取物所用原料在我国来源广,生产工艺比较简易、成本低,对人畜安全性好,对环境无污染,并且有花的清香,有效期可达一年以上。本发明中的复合植物提取物为食品级生物制剂,不仅挥发性好,用量少,易排除,在限量范围内使用对人体无害。
Description
技术领域
本发明涉及一种复合植物提取物及其制备方法与在抑制赭曲霉生长中的应用。
背景技术
植物提取物是一类植物源次生代谢物质,分子量较小、可随水蒸气蒸出、具有一定气味的挥发性油状物质。植物提取物具有抑菌、杀虫、抗氧化等活性,在医药、农药、饲料添加剂、食品防腐保鲜等方面有广泛应用。植物提取物具有来源于天然、对人体相对安全、与环境友好、生物活性多样等特点,越来越引起人们的关注。抗菌活性较高的植物提取物主要是醇、醛、酮以及酚类化合物,如百里香酚、肉桂醛、丁香酚、香紫苏醇、柠檬醛、香茅醛、香茅醇、大蒜素、姜黄素等。
赭曲霉(Aspergillusochraceus)是属于散囊菌目发菌科曲霉属的一种真菌,主要污染大米、小麦、燕麦、咖啡及其饮料等,赭曲霉污染可导致上述食品霉变变质,食品营养品质和加工品质大幅度下降,造成巨大的经济损失;更为严重的是赭曲霉在生长过程中可产生具有强毒性和致癌性的赭曲霉毒素,赭曲霉毒素有7种结构类似的化合物,其中赭曲霉毒素A(OTA)毒性最强,分布最为广泛。在已发现的真菌毒素中,赭曲霉毒素A(OTA)的危害性仅次于黄曲霉毒素,毒理学研究表明,OTA主要损害肝脏和肾脏,具有很强的肝脏毒性和肾脏毒性,大量的毒素也可能引起肠粘膜炎症甚至坏死;OTA被认为是目前致癌力最强的天然物质之一,被IARC定为二级致癌物。
由于赭曲霉毒素A(OTA)的毒性,国内外都公布了食品和饲料中OTA的限量标准,以规范生产,控制OTA的污染,最大限度地保障人和动物的食用安全,表1列举了欧盟、中国及台湾地区赭曲霉毒素的限量标准。
表1欧盟、中国及中国台湾对食品中OTA先行的限量标准
因此,筛选和开发能够高效抑制赭曲霉生长安全、无毒的天然活性物质,用于防控食品受到赭曲霉及赭曲霉毒素的污染,提升食品安全水平,具有十分重要的经济价值和社会意义。
发明内容
本发明的目的是提供一种复合植物提取物及其制备方法与在抑制赭曲霉生长中的应用。
本发明提供的一种复合植物提取物,它由肉桂醛、柠檬醛和丁香酚组成。
上述的复合植物提取物中,所述肉桂醛、所述柠檬醛和所述丁香酚的体积比可为2~6:2~6:1,具体可为2:2:1、2~4:2~4:1或2~5:2~5:1;
所述复合植物提取物的pH值可为6.5~7.0。
本发明中,采用食品安全级的碱性物质进行调节pH值,具体可为三乙醇胺或小苏打;
对pH值的调节是由于具体所采用的三种植物提取物占份额最多的是肉桂醛、柠檬醛占了复合植物提取物的80%,分子上的醛基有部份氢以H+解离,使复合植物提取物呈弱酸性,因此需用食品级的碱性物质调节pH至6.5-7.0,防止复合植物提取物酸碱基团对粮油原料及产品外表的侵蚀。
本发明还提供了上述的复合植物提取物的制备方法,包括如下步骤:将肉桂醛、柠檬醛和丁香酚按照用量由少到多的顺序依次加入混合,即得到所述复合植物提取物。
本发明中,所述肉桂醛、所述柠檬醛和所述丁香酚按照用量由少到多的顺序依次加入,是为了混合更加均匀;如果其中两种原料的量一样,顺序无先后。
上述的方法中,所述方法中还包括混合之后调节pH值的步骤。
本发明还进一步提供了上述的复合植物提取物在抑制赭曲霉生长中的应用。
上述的应用中,所述赭曲霉为赭曲霉(Aspergillusochraceus)CGMCCNo.3.4412;
所述复合植物提取物抑制如下产品中所述赭曲霉生长:谷物、食品和/或饮料;
所述复合植物提取物占所述产品的浓度可为100~500μL/L,具体可为150μL/L、200μL/L、250μL/L、150~250μL/L、150~200μL/L、200~250μL/L。
上述的应用中,采用熏蒸的方法处理所述产品;
所述谷物和食品的水分含量均可为0~15%,具体可为12%、15%、12~15%或5~15%。
上述的应用中,所述熏蒸处理的温度可为-10~37℃,具体可为28℃、-10~28℃、28~37℃或10~35℃;
所述熏蒸处理的时间可为3~45d,具体可为14d、3~14d、14~45d或10~25d;
所述熏蒸处理在密闭系统中进行。
上述的应用中,所述谷物为小麦、大麦、大米、玉米、黑麦和燕麦中的至少一种;
所述食品为咖啡豆、花生、核桃、开心果、榛子、腰果、杏仁、葵花籽、鲜葡萄和葡萄干中的至少一种;
所述饮料为葡萄汁、葡萄酒、咖啡饮品、麦芽饮料、酒精饮料、燕麦饮料和大米饮料中的至少一种。
本发明更进一步提供了一种赭曲霉生长抑制剂,该抑制剂的活性成分所述复合植物提取物。
本发明具有以下优点:
1、本发明复合植物提取物各组分肉桂醛、柠檬醛、丁香酚的原料资源在我国来源广,生产工艺比较简易、成本低,对人畜安全性好,对环境无污染,并且有花的清香,有效期可达一年以上。
2、磷化氢类化学抑菌剂,虽然挥发性良好但难除尽,并且易燃,严重威胁操作人员的人身安全和身体健康;而本发明中的复合植物提取物为食品级生物制剂,不仅挥发性好,用量少,易排除,在限量范围内使用对人体无害。
3、本发明所用的熏蒸剂为复合植物提取物,与单种植物提取物相比,效果更为显著且不易产生抗药性。
附图说明
图1为不同浓度复合植物提取物对赭曲霉生长的影响。
图2为复合植物提取物熏蒸对小麦(水分含量12%)中赭曲霉生长的抑制效果。
图3为复合植物提取物熏蒸对小麦(水分含量15%)中赭曲霉生长的抑制效果。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明,均为常规方法。
下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
下述实施例中,使用的赭曲霉(Aspergillusochraceus)购于CGMCC,该菌株在CGMCC的编号为3.4412;在本文的发明内容和权利要求中均表示成CGMCCNo.3.4412。
实施例1、复合植物提取物熏蒸抑菌效果测定
复合植物提取按照如下方法制备:
(1)植物提取物混合液的配制:按照体积比为4:4:2的比例取肉桂醛:柠檬醛:丁香酚;按照用量由少到多的比例依次混合三种植物提取物,边加边搅拌,混合均匀获得复合植物提取物;
(2)上述复合植物提取物所用原料植物提取物纯度均较高,无需除杂;
(3)复合植物提取物的pH调节:用三乙醇胺调节步骤(1)的复合植物提取物混合液的pH至6.5-7.0,即得到复合植物提取物。
PDA培养基:去皮马铃薯200g煮沸30min后取滤液,加入葡萄糖20g、琼脂16g,用蒸馏水定容至1000mL,加去离子水补足1L,115℃高压灭菌30分钟,降温到55℃左右倒平板,每平板20mL。
MEA培养基:酵母提取物(1g/L)、麦芽提取物(1g/L)、蛋白胨(1g/L)、KH2PO4(1g/L)、MgSO4·7H2O(0.3g/L)、KCl(0.3g/L)、ZnSO4·7H2O(0.05g/L)、CuSO4·5H2O(0.01g/L)、果糖(20g/L)、琼脂(15g/L),加去离子水补足1L,121℃高压灭菌20分钟,降温到55℃左右倒平板,每平板20mL。
菌种活化:把赭曲霉接种于PDA培养基上,28℃下生长14天,4℃储藏。
赭曲霉孢子液制备:用无菌棉签挑取平板培养基上的赭曲霉孢子溶于0.01%吐温20中,用无菌水稀释初始孢子液至孢子浓度为1×105cfu/mL。
把赭曲霉菌悬液10μL(1×105cfu/mL)接种于MEA培养基中央至完全吸收。将3片直径为10mm的灭菌滤纸片对称放置于培养皿盖的边缘,复合植物提取物分别1、2、3、4、5μL于不同培养皿中,使之均匀吸附于滤纸片上,空白不添加复合植物提取物,使复合植物提取物的浓度范围20-100μL/L,每个做3个平行,培养皿用封口膜密封后放入自封袋中,置于培养箱中28℃培养,培养20天后把没有赭曲霉生长的平板中取出复合植物提取物后再培养7天,观察有无菌丝的生长。每2天测量一次菌落直径,采用十字交叉法测定两次,结果取平均值。结果表明,所有熏蒸浓度的复合植物提取物均能显著抑制赭曲霉的生长,抑制效果随着复合植物提取物浓度的升高而增强,当复合植物提取物熏蒸浓度达到60μL/L时即能完全抑制赭曲霉的生长。取出复合植物提取物纸片并在通风橱内通风散味后,在放到培养箱培养7天,仍未见菌丝生长,表明赭曲霉孢子已被完全杀死。
实施例2、复合植物提取物熏蒸抑制赭曲霉在小麦(水分含量为12%)上的生长复合植物提取物同实施例1。
菌种活化和赭曲霉孢子液制备方法同实施例1。
小麦样品经1%次氯酸钠溶液清洗消毒,无菌水冲洗,无菌滤纸吸干表面水分。根据与预调目标水分差值加入定量无菌水,放于4℃静置24h使水分含量平衡,获得实验所需水分含量(12%)的小麦。将小麦以200g为单位分装到1L锥形瓶中,加入1mL孢子悬液(浓度2×106个/mL),轻轻摇动瓶子使孢子均匀覆盖到小麦上。紧接着滴加肉桂醛于无菌滤纸上,放入锥形瓶内,使肉桂醛添加浓度分别为0、50、100、150和200μL/L,密封后恒温恒湿箱28℃培养14d,测定污染率。每组做3个重复。结果表明,与对照相比,所有实验浓度复合植物提取物对小麦中赭曲霉污染率均有显著的降低效果,并且随着复合植物提取物浓度的升高而增强,当复合植物提取物浓度达到150μL/L时小麦污染率降为0,表明150μL/L的复合植物提取物能够完全抑制赭曲霉对小麦的侵染。
实施例3、复合植物提取物熏蒸抑制赭曲霉在小麦(水分含量为15%)上的生长
复合植物提取物同实施例1。
菌种活化和赭曲霉孢子液制备方法同实施例1。
小麦样品经1%次氯酸钠溶液清洗消毒,无菌水冲洗,无菌滤纸吸干表面水分。根据与预调目标水分差值加入定量无菌水,放于4℃静置24h使水分含量平衡,获得实验所需水分含量(15%)的小麦。将小麦以200g为单位分装到1L锥形瓶中,加入1mL孢子悬液(浓度2×106个/mL),轻轻摇动瓶子使孢子均匀覆盖到小麦上。紧接着滴加复合植物提取物于无菌滤纸上,放入锥形瓶内,使复合植物提取物添加浓度分别为0、100、150、200和250μL/L,密封后恒温恒湿箱28℃培养14d,测定污染率。每组做3个重复。结果表明,与对照相比,所有实验浓度复合植物提取物对小麦中赭曲霉污染率均有显著的降低效果,并且随着复合植物提取物浓度的升高而增强,当复合植物提取物浓度达到200μL/L时小麦污染率降为0,表明200μL/L的复合植物提取物能够完全抑制赭曲霉对小麦的侵染;水分含量15%小麦的赭曲霉污染率明显高于12%的小麦。
Claims (10)
1.一种复合植物提取物,其特征在于:它由肉桂醛、柠檬醛和丁香酚组成。
2.根据权利要求1所述的复合植物提取物,其特征在于:所述肉桂醛、所述柠檬醛和所述丁香酚的体积比为2~6:2~6:1;
所述复合植物提取物的pH值为6.5~7.0。
3.权利要求1或2所述的复合植物提取物的制备方法,包括如下步骤:将肉桂醛、柠檬醛和丁香酚按照用量由少到多的顺序依次加入混合,即得到所述复合植物提取物。
4.根据权利要求3所述的方法,其特征在于:所述方法中还包括混合之后调节pH值的步骤。
5.权利要求1或2所述的复合植物提取物在抑制赭曲霉生长中的应用。
6.根据权利要求5所述的应用,其特征在于:所述赭曲霉为赭曲霉(Aspergillusochraceus)CGMCCNo.3.4412;
所述复合植物提取物抑制如下产品中所述赭曲霉生长:谷物、食品和/或饮料;
所述复合植物提取物占所述产品的浓度为100~500μL/L。
7.根据权利要求6所述的应用,其特征在于:采用熏蒸的方法处理所述产品;
所述谷物和食品的水分含量均为0~15%。
8.根据权利要求7所述的应用,其特征在于:所述熏蒸处理的温度为-10~37℃;
所述熏蒸处理的时间为3~45d;
所述熏蒸处理在密闭系统中进行。
9.根据权利要求6-8中任一项所述的应用,其特征在于:所述谷物为小麦、大麦、大米、玉米、黑麦和燕麦中的至少一种;
所述食品为咖啡豆、花生、核桃、开心果、榛子、腰果、杏仁、葵花籽、鲜葡萄和葡萄干中的至少一种;
所述饮料为葡萄汁、葡萄酒、咖啡饮品、麦芽饮料、酒精饮料、燕麦饮料和大米饮料中的至少一种。
10.一种赭曲霉生长抑制剂,其特征在于:该抑制剂的活性成分为权利要求1或2所述的复合植物提取物。
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CN107711834A (zh) * | 2017-10-23 | 2018-02-23 | 天津农学院 | 一种含有丁香酚、肉桂醛和柠檬醛天然复配杀菌剂 |
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CN108029762B (zh) * | 2017-11-29 | 2021-12-03 | 中国农业科学院农产品加工研究所 | 复合植物提取物及其在粮食储藏防霉中的应用 |
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