CN104824119A - 栗单宁提取物作为抗氧化剂、抗微生物添加剂的用途和降低亚硝胺和真菌毒素的用途 - Google Patents
栗单宁提取物作为抗氧化剂、抗微生物添加剂的用途和降低亚硝胺和真菌毒素的用途 Download PDFInfo
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- CN104824119A CN104824119A CN201510183812.9A CN201510183812A CN104824119A CN 104824119 A CN104824119 A CN 104824119A CN 201510183812 A CN201510183812 A CN 201510183812A CN 104824119 A CN104824119 A CN 104824119A
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Abstract
描述了单独和/或与其他多酚混合的栗单宁提取物(欧洲栗)和/或其级分作为抗氧化剂、抗微生物添加剂的用途和在食品原料和人类和动物食品中降低亚硝胺和真菌毒素的用途。
Description
本发明涉及栗单宁提取物(ches tnut tannin extract)作为抗氧化剂、抗微生物添加剂的用途和用于降低亚硝胺和真菌毒素的用途;更特别地,本发明涉及单独或与其他多酚混合的栗单宁提取物(欧洲栗)和/或其级分作为抗氧化剂、抗微生物添加剂的用途和在食品原料和人类食品和动物食品、微生物负荷(microbial charge)中降低亚硝胺和真菌毒素的用途和在食品原料和人类和动物食品中降低形成亚硝胺和真菌毒素的风险的用途。
本发明源自记载在2012年8月8日提交的专利申请号为MI2012A001419、名称为“针对降低亚硝胺含量的烟草生产方法”(“Method for the productionof tobacco aimed at reducing the content of nitrosamines”)中得到的先前研究的继续。
氧化链反应引起食品原料和终端食物产品中的氧化变质。活性氧(ROS)通过酶的、化学的、光化学的、和辐照的方式形成,并且通过相同ROS之间的分解和反应形成。羟基自由基和氧是最有活性的ROS。它们与食品物质的反应产生不期望的和致癌的挥发性化合物,其破坏必需的营养元素和改变蛋白质、脂类和碳水化合物的功能性质。例如,脂类的氧化产生低分子量和挥发性物质,例如醛类、醇类和酮类。可能会发生蛋白质的交联或裂解,同样就碳水化合物、具有羧基的低分子量化合物产物而言也会发生交联或裂解。维生素容易被ROS氧化,特别是β-胡萝卜素以及维生素E、维生素B2、维生素D和维生素C[ChoeE.,Min D.B,Chemistry and Reactions of Reactive Oxygen Species in Foods,Crit.Rev.In Food Sci.Nutr.46(1),1-22,2006]。这导致感官和营养质量上的损失,和产生异常气味和香味。
总的微生物负荷,特别是大肠杆菌L(Escherichia coli L.),沙雷菌属(Serratia sp.),沙门菌属,总和粪大肠菌群,酵母菌和霉菌,是引起食物特别是肉类和乳制品的微生物污染的主要因素[Jay J.M.,Modern foodmicrobiology,ISBN 0-412-07691-8,1996;Zhno G.,Giuffreda A.,GiufrèN.,Greco V.,Panebianco A.,Accertamenti su Serratia marcescensisolata da carne suina cotta e refrigerata,Industrie Alimentari 49(508):15-31,2010;Yang X.,Badoni M.,Youssef M.K.,Gill C.O.,Enhanced controlof microbiological contaminat ion of product at a large beef packing plant,J.Food Prot.75(1):144-9,2012;D’Elia G.,Bacci C.,Bassi L.,BoniE.Alpigiani I.,Brindani F.,Le carni separate meccanicamente:aspettiproduttivi microbiologici e di etichettatura,Industrie Alimentari51(521):30-36,2012]。
N-亚硝胺类相应地通过亚硝酸盐或亚硝酸与仲胺和叔胺在酸环境和合适的条件下的反应形成[Gray J.I.e Dugan L.R.Jr.,J.Food Sci 40(5),981-984,1975]。亚硝酸盐和硝酸盐被慎重地添加至咸肉中以稳定其颜色并提供对保妥适(Botox)风险的防护;此外,硝酸盐包含在很多蔬菜中,在此硝酸盐可被还原为亚硝酸盐。胺及其前体,例如蛋白质,氨基酸,磷脂和季铵化合物也存在于很多蔬菜和动物来源的食物产品中。亚硝胺长期以来都被认为是致癌化合物(IARC Monographs 1974,vol.4),并且其浓度的降低是用于人类和动物的食物产品卫生和安全的主要目的。被认为是主要的亚硝胺类物质有,作为纯粹举例并且不排除其他物质,衍生自亚硝酸盐或亚硝酸与氨基醇类,二乙醇胺,三乙醇胺,N,N-二乙醇胺,N,N-二甲基乙醇胺,1-甲氧基-2-丙胺,2-氨基-2-甲基-1-丙醇,3-甲氧基丙胺,吗啉的反应的亚硝胺;本发明并不仅限于这些,并且考虑相似的所有存在于食物产品中的亚硝胺,不管是新鲜的还是通过各种机理最近形成的,在其保存期间,在用于人类和动物的营养的最终产品生产之前和之后并且直到消费为止。
真菌毒素,是具有毒性活性的真菌代谢物,能够相应地导致急性和/或慢性中毒的发生,也称为真菌中毒。它们在动物饲料中的存在会导致降低生长和繁殖的问题,并且在更严重的情况下,会导致饲养动物的急性中毒。此外使用受污染的食物产品,不仅在动物摄取它们上有负面影响,而且导致毒素或其衍生物进入牛奶,鸡蛋和肉类[Miele S.,Salera E.,Sbrana M.,2004.LeMicotossine:linee guida per il controllo agronomico e identificazioneanalitica.Seminario Regione Toscana-Accademia dei Georgofili,26October 2004,atti]。在攻击谷类的真菌毒素中,镰刀菌属(Fusarium)确实被最频繁地检测出来。最主要的镰刀菌毒素(fusariotoxins)是脱氧瓜萎镰菌醇(DON),玉米赤霉烯酮(ZEA),瓜萎镰菌醇(NIV)(所有三种物质通过黄色镰刀菌(F.culmorum)和禾谷镰刀菌(F.graminearum)产生),伏马菌素类(通过轮枝镰刀菌(F.verticillioides)(念珠形)和串珠镰刀菌(F.proliferatum)产生),毒素T2和HT-2(通过拟枝孢镰刀菌(F.sporotrichioides)产生),和蛇形菌素(DAS)(通过梨孢镰刀菌(F.poae)和木贼镰刀菌(F.equiseti)产生)。
自从开始关注这些毒素的临床反应后,人们怀疑伏马菌素B1能够导致食道癌。马和猪被证明对伏马菌素类特别敏感。在马身上,这些毒素导致神经问题(毒性脑膜炎),其中在猪身上,它们导致胸膜肺炎(肺水肿)。在牛身上,伏马菌素类导致肝炎和免疫系统的降低(免疫抑制)。单端孢霉烯类(DON,DAS,NIV,T2)通常导致动物健康方面的渐进性衰退,伴随着产量的降低并且因此的重大经济损失。它们同时带来的综合征可以被辨识为催吐,以呕吐和出血为特征(Viljoen J.H.,Mycotoxins in grain and grain products in South Africaand proposals for their regulations,Univ.of Pretoria etd,PhD论文,2003)。
黄曲霉毒素类(AF)是黄曲霉(Aspergillus flavus)(AF B1和B2的生产者)和寄生曲霉(Aspergillus parasiticus)(AF B1,B2,G1和G2的生产者)的菌株产生的一组真菌毒素。在大多数情况下,AFB1是以最高量存在的毒素并且由于其高的急性和慢性毒性和对动物的致癌活性,研究人员对其特别感兴趣,此外其对人类还有潜在的影响。在所有研究过的动物种类中,AFs导致肝癌,并且同时也导致肾癌;特别地,AFB1已知具有最强的口服致肝癌活性。其摄取能够通过受污染的蔬菜食物产品或动物产品发生,例如牛奶。实际上奶牛,将AFB1(1=对人类来说致癌)转换成AFM1,其IARC具有2B的分级,其意味着“对人类来说可能致癌”。
最后,赭曲霉毒素类在我们的周围通过青霉菌(Penicillium spp.)和赭曲霉菌(Aspergillus ochraceus)产生。赭曲霉毒素类对人类极其有害;特别地,赭曲霉毒素A(OTA)能够在肾和免疫系统方面导致严重的问题,并且在过去被认为会导致肿瘤和婴儿畸形。这种物质可能存在于有机体中并且导致长期损害。其能够通过受污染的蔬菜食物产品或衍生自谷类的产品被摄取,例如用大麦芽生产的啤酒的情况[EFSA,Statement on recent scientific informationon the toxicity of Ochratoxin A,EFSA J.8(6):1626,2010]。
背景技术
因此消费者倾向于要求更简单的并且“绿色”标签的食物产品。因此,在此方面,人们对从食物产品中排除合成防腐剂例如TBHQ(叔丁基-羟基-醌)、BHT(丁基-羟基-甲苯)、BHA(丁基-羟基-茴香醚)和EDTA(乙二胺四乙酸)越来越感兴趣。代替这些合成防腐剂,对天然物质存在增长的需求,这些天然物质特别是唇形科提取物,例如迷迭香,鼠尾草,牛至,百里香,薄荷和它们的混合物,或者迷迭香酸和鼠尾草酸[Sandusky C.L.,Reynhout G.S.,JonesT.S.,Method of extending color life of modified atmosphere packagedfresh red meat using Labiatae plant extracts,US Pat.7,037,543/2000]。
已知食物-单宁化合物很少结合使用,仅仅是使用浓缩的单宁。MauroSaviola Group的名称为“单宁和非单宁材料的天然提取物在改进土壤肥力和提供培育起始效果的用途和单宁和非单宁植物组分”(“Use of natural extractsof tannin and non-tannin materials for improving soil fertility andproviding a starter effect on cultivations,and a tannin and non-tanninphytocomposition”,)的专利申请EP2345628(A1)已经描述了获自栗属(Castaneaspp.)的提取物。
因此即使为了其他目的,在农业生产中,也存在已经使用的产品,例如为了调整土壤和灌溉水的pH,和作为生长促进剂和抗各种害虫,基于SaviolaGroup申请的专利申请EP1464635A1(2003年08月26日)和BP2345628A1(2011年01月14日)。
本专利申请为上述提到的专利申请的继续和发展,以达到所有效果。
发明详述
本发明克服了现有技术中的问题,出乎意料地发现一种处理用于人类和动物的食物原料和食物产品的方法,其基于具有抗氧化剂、抗微生物和抗真菌特性的天然栗单宁植物提取物(欧洲栗)及其标准化级分的应用,下面进行更加详细的描述。
本发明是先前研究成果的继续,即已经提到的,在2012年8月8日提交的专利申请号为MI2012A001419、名称为“针对降低亚硝胺含量的烟草生产方法”的专利申请。
已知栗单宁提取物以可水解的单宁的存在为特征,其主要由酚化合物组成,例如没食子酸和鞣花酸,其部分或全部被D-葡萄糖分子酯化。该水性提取物的抗氧化剂活性已经被证实并且可以用FRAP检测法来测量(在三吡啶基三嗪复合物中还原成蓝色亚铁形式,在593nm吸光度增加)。
出乎意料地发现了本发明的目的即栗单宁提取物在用于人类和动物的食物原料(例如油、油菜籽、谷类)或食物产品中的使用抑制了这些材料的氧化降解现象并且同时降低微生物污染和亚硝胺和真菌毒素的形成,并且当使用合适的浓度,还与其他多酚混合时,没有改变味道和香味。
因此,本发明的一个目的涉及单独或与其他多酚混合的栗单宁提取物和/或其级分作为抗氧化剂、抗微生物添加剂的用途和在食品原料和人类和动物食品中降低亚硝胺和真菌毒素的用途。
当栗单宁提取物和/或其级分与其他多酚混合使用时,所述多酚优选选自绿茶提取物,朝鲜蓟提取物,橄榄树叶,橄榄酱提取物,葡萄籽提取物,迷迭香提取物,甜菜根提取物。所述多酚对栗单宁提取物的抗氧化剂和抗微生物效果具有协同作用。当添加至人类或动物食物原料和食物产品中时,这些提取物进一步降低亚硝胺和真菌毒素的含量。
本发明进一步的目的还涉及一种处理用于人类和动物的食物原料和食物产品的方法,其包括添加单独和/或与其他多酚混合的栗单宁提取物和/或其级分特别地用于降低谷类中的真菌毒素的含量,降低豆类中的真菌感染,用于烘焙产品的稳定,肉类的防腐和动物饲料的稳定及其生物和功能性能的增加。
为了本发明的目的,但并不限于其他应用的可能性,栗单宁提取物和/或其级分以液体的形式使用,其中相对于提取物或其级分的总重量,单宁的含量为1-40wt%,优选13-25wt%,或以粉末的形式使用,其中相对于提取物或其级分的总重量,单宁的含量为30-90wt%。
下面描述所述栗单宁提取物和/或其级分的获得方式。
术语栗单宁提取物的含义为液体和固体形式的下述定义的级分1-9的任意重量比的任意混合物。
这些产品可以添加:1)到种子和原料(例如,并不限制其他应用的可能,谷类和proteoleaginose种子)中,以在播种阶段,在田地里生长期间和在收获和储存阶段中避免真菌种类的污染现象和接下来的降解;2)到用于食品工业的原料(例如,并不限制其他应用的可能,油和蛋白质饼)中,用来降低氧化现象和接下来的腐败以及微生物污染;3)到用于动物和人类的处于生产或配方阶段的食物中,以保持其质量和营养特性并降低健康风险。
在种子的情况下,处理方法已经发展为包括如下实现的两种应用:第一,在种子上(例如,并不限制其他应用的可能,小麦籽粒);第二,在玉米穗上。第一种应用以使用相对于要处理的种子的总重量的0.1-10wt%且优选相对于要处理的种子的总重量的0.3-4wt%浓度的粉末形式的单宁提取物来实现,其中所述粉末形式的单宁提取物的单宁含量为30-90wt%。
第二种应用以使用相对于进行玉米穗喷雾处理(通常约100kg/ha)所使用的水或液体工具(liquid means)的总重量的0.5-5wt%且优选1-1.5%浓度的液体形式的单宁提取物或其一种级分来实现,其中所述液体形式的单宁提取物或其一种级分的单宁含量为13-25wt%。特别地,使用下述实施例1中描述的级分4。
在食品原料的情况下,例如加入单宁含量为13wt%的栗提取物至谷粒和proteoleaginose种子中,相对于要处理的产品的总重量,提取物的浓度为0.01-5wt%,并且优选0.1-0.5wt%。该产品在收获阶段中或储存低点的谷物再混合阶段中喷洒在谷粒和种子上。
在饲料产品的情况下,制备宠物食品的配方颗粒,向其中添加37-40wt%单宁的液体栗提取物,以增加它们的保存期和降低微生物污染的风险,以及接下来的动物饮食问题和腹泻的风险。提取物在挤出之前加入混合物,相对于要处理的产品的总重量其用量为0.01-5wt%,优选0.3-0.7wt%。
在终端食物产品的情况下,同时使用单独或与其他多酚混合的栗单宁提取物和其级分,其中所述多酚特别是绿茶提取物,羟基酪醇,朝鲜蓟提取物,橄榄树叶,橄榄酱提取物,葡萄籽提取物,迷迭香提取物,甜菜根提取物(Betavulgaris L.var.cruenta L.Salisb.)。所检查的终端产品为鲜肉,咸肉(例如香肠)和烘焙产品,同时与选择的多酚混合的单宁提取物和/或其级分的剂量相对于要处理的产品的总重量为0.0001-3wt%,优选0.001-1.5wt%。
特别地,本专利申请的目的即栗单宁提取物及其级分通过栗木植物的热水提取和接下来冷分级来获得。更特别地,冷提取和分级方法包括如下阶段:
a)借助浸提的栗木的热水提取阶段;
b)来自步骤a)的浸提液的预过滤阶段,使用珍珠岩过滤器以除去粗颗粒(锯末、土壤、沙子等),然后使用过滤器芯保留具有大于25微米尺寸的颗粒;得到的包含4-5wt%单宁的液体级分被称为“过滤后的单宁汤(broth)”的单宁(以下用级分1表示过滤后的单宁汤)。
c)第一次浓缩步骤,通过来自步骤b)的液体的纳滤进行;阶段c)结束后得到的产品为:“第一浓缩步骤渗透物”(级分2)和“第一浓缩步骤浓缩物”(级分3),后者具有约13wt%的单宁含量;
d)阶段c)得到的“第一浓缩步骤浓缩物”产品通过下述依次实施的步骤来沉降和净化:
-在10-12℃冷却24-48小时,使仍然存在的待沉降的粗颗粒沉降;
-机械离心分离以获得没有悬浮颗粒的液体,其被称为“冷却后的第一浓缩步骤浓缩物”(级分4);级分4的单宁含量保特在约13wt%单宁;
沉降和净化(通过离心分离)步骤的残余材料被称作“净化沉降物”(级分9);
e)借助纳滤进行最终浓缩,结束后获得“第二浓缩步骤渗透物”(级分5)和“第二浓缩步骤浓缩物”(级分6),后者具有约40wt%的单宁含量;
反渗透步骤,以最优化来自步骤c)和e)的渗透物的单宁的回收工艺:将该两种渗透物(级分2和级分5)合并并且通过反渗透进行处理;在该阶段结束后,得到“反渗透渗透物”(级分7)和“反渗透浓缩物”(级分8),后者具有约6-7wt%的单宁含量。通过浸提的提取阶段a)在110-120℃的温度下实施,栗木碎片装入逆流、串联操作的一连串的高压釜中。假设有一组8个高压釜,干净的水在110-120℃的温度下进入高压釜组,与几乎用尽的木头(即已经经历了6次用水溶液提取步骤的木头)相遇,并且该富含单宁的溶液离开所述第一高压釜,进入临近的高压釜(第二高压釜),在所述第二高压釜中其溶解以较高浓度存在于木头中的单宁,所述木头已经依次经历五次水溶液提取。离开所述第二高压釜的进一步富含单宁的溶液,溶解在临近高压釜(第三高压釜)中的木头中的单宁,所述第三高压釜包含的木头已经经历了四次水溶液提取。如此等等,直到特别富含单宁的溶液离开倒数第二个高压釜(第六个高压釜)与新木头相遇,即进入一连串高压釜中的新鲜破碎的木头,然后从所述高压釜(第七高压釜)中排出,形成未过滤的单宁汤,其送入后续处理步骤。
所述来自浸提提取的溶液的特征在于4-5wt%的单宁百分比,和约60-65%的提取产率。
特别地,从上面描述的工艺中得到的级分如下:
1、过滤后的单宁汤;
2、第一浓缩步骤渗透物;
3、第一浓缩步骤浓缩物(约13wt%的单宁);
4、冷却后的第一浓缩步骤浓缩物(约13wt%的单宁);
5、第二浓缩步骤渗透物;
6、第二浓缩步骤浓缩物(约40wt%的单宁);
7、反渗透渗透物;
8、反渗透浓缩物;
9、净化沉降物;
用于根据本发明的用途的优选级分是如下级分:
4、冷却后的第一浓缩步骤浓缩物(约13wt%的单宁);
6、第二浓缩步骤浓缩物(约40wt%的单宁);
所述级分单独使用和/或与彼此结合使用和/或与其他级分结合使用和/或与其他多酚混合使用。
所述级分可以以上述公开的方法直接获得的浓缩液体的形式使用,或者可以以雾化后粉末的形式使用。
该雾化阶段通过将液相进料至喷雾干燥器中来实施,以此获得粉末形式的产品。
使用上述方法获得的根据本发明的栗单宁提取物的应用特别有效,与已知的单宁提取物相比,该栗单宁提取物的特征在于高得多的抗氧化剂作用和食物产品中存在的细菌总数、总和粪大肠菌群、酵母菌和霉菌的大幅下降。
下列实施例仅仅是为了描述本发明的目的,不能认为是对本发明的保护范围的限定,本发明的保护范围如权利要求所限定。
实施例1
可用级分的抗氧化剂和抗自由基性能
分析的样品如上面所述获得,其来自栗木植物的热水提取和冷级分。
特别地,所检测的下述级分为:
1、过滤后的单宁汤;
2、第一浓缩步骤渗透物;第一浓缩步骤浓缩物(约13wt%的单宁);
3、冷却后的第一浓缩步骤浓缩物(约13wt%的单宁);
4、第二浓缩步骤渗透物;
5、第二浓缩步骤浓缩物(约40wt%的单宁);
6、反渗透渗透物;
7、反渗透浓缩物;
8、净化沉降物。
这些级分的抗氧化剂和抗自由基活性显示在表1中,后者使用稳定自由基DPPH方法在体外测量(Campo M.,Pinelli P.,Romani A.,HPLC-DAD-MSCharacterization and Antioxidant Activity of Sweet Ches tnut(Castaneasativa Mill.)Fractions,XXVI International Conference of Polyphenols,Polyphenols Communication,2012,Vol.I,Pag.135-136;FOOD CHFMISTRY,2014年提交;Campo M.,Pinelli P.,Romani A.,HPLC-DAD-ESI-MSCharacterization and Antioxidant Activity of Hydrolyzable Tannins fromSweet Chestnut(Castanea sativa Mill.)Fractions.FOOD CHEMISTRY,2014年提交)。
表1:抗氧化剂和抗自由基活性,其表达为没食子酸(GAE)的当量和DPPH的初始活性降低至50%时所需要的抗氧化剂的量(EC50)
根据实施例1的加工级分 | GAE(g/100g提取物) | EC50(μM) |
1 | 2.4 | 0.70 |
2 | 0.6 | 2.00 |
3 | 9.1 | 0.44 |
4 | 7.9 | 1.00 |
5 | 2.4 | 1.43 |
6 | 31.0 | 2.56 |
7 | 0.1 | 2.40 |
8 | 7.4 | 1.51 |
9 | 0.5 | 0.55 |
实施例2
为了控制拟茎点霉的大豆种子的处理[f.asc.Diaporthe phaseolorum var.sojae e var.culivora(大豆茎溃疡剂)]
大豆种子被拟茎点霉/茎腐病污染使得同样的种子不能再作为合格的种子应用,除非使用化学单宁物质进行处理。在大豆种子上进行了总共10个应用测试,以减缓高污染。出乎意料地发现以相对于要处理的种子的总重量的0.30wt%的浓度使用13wt%单宁的栗提取物(级分4),通过在装袋之前的处理阶段喷洒该产品至种子上,可以减少种菌的存在量至检测限之下。
使用根据本发明的配方处理种子达到控制真菌孢子的存在和活力的目的,使用由Zhang A.W.et al.[Molecular detection of Diaporthe phaseolorumand Phomopsis longicolla from soybean seeds.Phytopathology 89:796-804,1999]和Schena L.et al.[Real-time quantitative PCR:a new technology todetect and study phytopathogenic.European Journal of Plant Pathology110:893-908,2004]所描述的方法。
实施例3
为了降低伏马菌素类和脱氧瓜萎镰菌醇(DON)的含量,使用栗单宁提取物处理玉米的种子和穗
两年预备测试之后(其为仅用粉末形式的单宁提取物处理硬粒小麦种子或仅用单宁提取物溶液或其级分处理玉米穗),开发了一种方法,其由连续起作用的两种协同应用组成。第一种处理由小麦籽粒的单宁处理组成,在马上播种前进行,使用粉末形式的栗单宁提取物,其包含75wt%的单宁,以0.5-1.5kg/100kg籽粒的浓度实施。第二种处理通过应用来表示,通过叶子,在实施例1的级分4的阶段50Zadoks级别,以在水中的1-1.5wt%的浓度实施,以1001/ha的剂量分布。接下来对收集的籽粒样品的分析显示:相对于没有处理的空白样品,根据新方法的处理显著地降低了玉米穗的真菌腐烂的程度并将谷粒的来自伏马菌素和DON的污染显著降低到法律强制限制以下(例如,在硬粒小麦的情况下,1750μg/kg的DON)。与空白样品相比,处理过的样品在收获时在籽粒中测量的总量和内生的真菌负荷更低,因此证明了通过介入自然产品,来自伏马菌素类和DON的污染降低,具有低的环境影响。
实施例4
基于单宁提取物及其级分与基质的混合物和基于作为食品成分/香料使用的多酚的提取物的配方
下述配方被配制并以各种测试方法进行测试:
1、具有15-90wt%单宁含量的栗单宁提取物粉末;
2、具有3-40wt%单宁含量的栗单宁提取物液体;
3、固体菜蓟属(Cynara)TAN 90/10:90wt%的固体菜蓟属提取物(喷雾干燥的朝鲜蓟26.94mg/g总多酚)和10wt%的具有75wt%单宁含量的栗单宁提取物粉末;
4、固体菜蓟属TAN 80/10/10:80wt%的固体菜蓟属提取物(喷雾干燥的朝鲜蓟26.94mg/g总多酚),10wt%的具有75wt%单宁含量的栗单宁提取物粉末和10wt%的浓缩自具有95%滴定度的葡萄籽粉末的单宁提取物;
5、液体菜蓟属TAN 85/15:85wt%的液体菜蓟属提取物(在蒸发器中浓缩至糊状,提取自干朝鲜蓟叶子)和15wt%的具有40wt%单宁含量(级分6)的栗单宁提取物液体;
6、液体菜蓟属TAN 80/15/5:80wt%的液体菜蓟属提取物(在蒸发器中浓缩至糊状,提取自干朝鲜蓟叶子),15wt%的具有40wt%单宁含量(级分6)的栗单宁提取物液体和5wt%的浓缩自具有95%滴定度的葡萄籽粉末的单宁提取物;
7、浓缩的TAN-OL糊50/50:50wt%的具有30wt%单宁含量的栗单宁提取物液体和50wt%的在橄榄苦苷中具有3%滴定度的浓缩的橄榄树叶糊;
8、浓缩的TAN-OL-RED糊30/30/40:30wt%的具有30wt%单宁含量的栗单宁提取物液体,30wt%的在橄榄苦苷中具有3%滴定度的浓缩的橄榄树叶糊和40wt%的甜菜根提取物;
9、浓缩的TAN-OHTy-RED糊20/40/40:20wt%的具有30wt%单宁含量的栗单宁提取物液体,40wt%的获自具有表达为羟基酪醇的3%滴定度的橄榄糊的浓缩溶液和40wt%的甜菜根提取物;
10、浓缩的TAN-OHTy-RED糊30/70:30wt%的具有30wt%单宁含量的栗单宁提取物液体和70wt%的获自具有表达为羟基酪醇的3%滴定度的橄榄糊的浓缩溶液;
11、浓缩的ROSMA-TAN糊70/30:70wt%的在迷迭香酸和鼠尾草酚中具有3%滴定度的迷迭香提取物和30wt%的具有30wt%单宁含量的栗单宁提取物液体;
12、Green-TAN-OL粉末20/40/40:20wt%的在EGCG中4-95%的绿茶提取物,40wt%的具有40-75wt%单宁含量的粉末形式的栗单宁提取物,和40wt%的在橄榄苦苷中具有3-17%滴定度的粉末形式的橄榄油提取物。
这些混合物适用于烘焙产品,乳制品和鲜肉和咸肉,具体细节如下述实施例描述。
实施例5
用于稳定烘焙产品的栗单宁提取物的应用
根据本发明的单独或与获自橄榄糊的羟基酪醇混合的栗单宁提取物具有抗氧化剂和抗微生物性质,用于稳定甜面包干、甜饼干和填充杏仁的饼干。使用TAN-OHTy配方,即前述实施例4中的配方10,与单独使用羟基酪醇相比较。
在甜面包干的测试中,TAN-OHTy以等于750ppm的浓度添加进生面团中。在该TAN-OHTy配方中栗单宁提取物具有30wt%的单宁含量,TAN-OHTy的浓度使得最终的单宁浓度达到300-5,000ppm,优选500-750ppm,在任何情况下能够用于本发明的目的。羟基酪醇以500ppm的浓度添加。评价感官特性并且对所有类型产品进行腐败测试,在时间零点和在烘箱中在52℃下加速老化后进行分析。
该加速老化方案与欧洲药典(EDQM参考标准)中描述的相一致,该方案模仿10天至一个月的稳定性。经过验证的内部食品方案确立了一个月的稳定性,相当于在52℃下7天的加速老化。
在时间0,70和110天时,对甜面包干的稳定性测试的结果显示在表2中。
从这些结果中可以看出,在加速老化方案的条件下,TAN-OHTy配方明显改善耐腐败的所有定性参数。对保持在室温下的样品进行类似的分析,在8个月和1年的防腐后的结果显示两种用天然提取物处理的样品没有变味并且显示出更低的过氧化物和总酸度值。相反,在18个月的室温下的稳定性分析后,空白样品显示出令人不愉快的腐败气味,然而在处理过的样品中不存在该气味。
这通过Kreis测试[Narasimhan S.,Vasanth Kumar A.K.,Ravi R.,ChandN.,Optimization of Kreis Test for edible oils,J.Food Lipids6(2):107-115,1999]得到确认,其结果非常明显地确认没有天然提取物的产品在该阶段存在氢过氧化物的分解,其中形成了可挥发产品类型的氧化。根据本发明配制的天然成分/香味证明确保了抗氧化剂保护,也作为自由基清除剂,干涉和减缓氢过氧化物的形成。
应指出经受加速老化测试(时间0)的样品的游离酸度不能被认为是有用的参数,因为它不是脂类级分氧化的指示,这是由于对产品经历的高温的依赖性。同样对于过氧化物的数量,相对于在室温下的对照加速的储藏寿命研究显示出第一老化方案也不是可靠的指示,因为可氧化的底物证明是更丰富的,这是由于甘油三酸酯的水解(与特级初榨橄榄油添加进产品中相关联),其发生在高温下,在所有的样品中过氧化物的数量增加,甚至通过天然抗氧化剂的存在也不能准确的限制该过程。
在此情况下,也证明了氧化和腐败过程在用TAN-OHTy处理的样品中被更多地限制。
在饼干、杏仁饼干和烤杏仁饼干中进行了相似的实验,其目的在于抑制氧化和腐败过程。特别地,使用了实施例4的配方3,4,5,6和12,在本实施例描述的测试中使用相同的剂量。
上面提到的所有烘焙产品都经历了感官测试,以正确调整使用的单宁产生的苦和涩的味道。填充饼干和杏仁饼干被证明比甜面包干更容易适应。
表2-“甜面包干产品”经受加速老化后的稳定性的结果:(1)空白样品。(2)羟基酪醇,(3)根据本发明的配方:TAN-OHTy。
实施例6
栗单宁提取物用于稳定基于肉的产品的应用
具有抗氧化剂和抗微生物性能的栗单宁提取物用于稳定猪肉,牛肉汉堡和香肠,使用前述实施例4中描述的各种配方。
在第一测试中,基于碎牛肉的产品用下述配方和剂量处理:
1)未处理的空白样品(对照);
2)标准添加剂1:100g抗坏血酸钠、100g抗坏血酸+10g胭脂红的混合物;210mg的该混合物添加至100g新鲜切碎的牛肉中;
3)固体菜蓟属TAN 80/10/10(实施例4的配方4),以等于300mg的用量添加至100g新鲜切碎的牛肉中;
4)TAN-OL-RED(实施例4的配方8),以等于300mg的用量添加至100g新鲜切碎的牛肉中;
5)TAN-OHTy-RED(实施例4的配方9),以等于300mg的用量添加至100g新鲜切碎的牛肉中。
对照涉及:pH和细菌总数(表3)并通过视觉检验和感官测试完成。
对细菌总数的对照(参见表3)显示了在4天之后,使用配方固体菜蓟属TAN 80/10/10和TAN-OHTy-RED处理的样品与使用标准添加剂1处理的样品相比显示出改进的表现。感官分析显示配方固体菜蓟属TAN 80/10/10具有好的表现和TAN-OHTy-RED的应用使得明亮的红色得到了显著的保护,其中涉及甜菜根提取物的所有红色颜料在样品的测试条件下均是稳定的。
第二测试对内脏杂碎,碎牛肉和猪肉香肠进行,涉及以内脏杂碎重量的1.2wt%,碎成犬牛肉(adult beef)的0.4wt%和香肠的0.2wt%的浓度,使用与实施例4相同的配方(wt%表示各自相对于内脏杂碎,碎牛肉和猪肉香肠的总重量)。在这种情况下,除细菌总数外,也评估了以MPN/100g计量的总大肠菌群、粪大肠菌群,以UFC/g计量的酵母菌和酵母菌+霉菌。
内脏杂碎和碎成犬牛肉处理后,酵母菌和霉菌证明被抑制,其中由于酵母菌和霉菌不存在香肠中,所以在香肠中没有对它们进行评估。
在内脏杂碎中,初始的总大肠菌群显示出等于1800UFC/g的值;在使用固体菜蓟属TAN 80/10/10的测试中,4天后其降低至1200UFC/g,随后在8天之后,升高至等于4400的值,而在使用TAN-OL-RED和TAN-OHTy-RED的测试中,在最初的4天后,该值降低至600UFC/g,在8天之后各自升高至1500和1600UFC/g。
在碎成犬牛肉中所测量相同的参数初始显示等于20的值,在菜蓟属TAN80/10/10的测试中,8天之后达到1600UFC/g,而在TAN-OHTy-RED的测试中,4天后达到90的值,其保持恒定直到第8天。
在使用固体菜蓟属TAN 80/10/10的香肠测试中,总大肠菌群在最初的4天里保持20和50UFC/g的值,在8天后所有样品均降低至20的值。
在香肠样品和碎成犬牛肉样品中均没有粪大肠菌群的存在,而未处理的内脏杂碎对照样品中发现在时间零点时有27,800MPN/100g(MPN/100g相当于最大可能数值/100g),并且第4天时达到110,000MPN/100g。
另一方面,在内脏杂碎的测试中该值为100,000MPN/100g,其中固体菜蓟属TAN 80/10/10在4天后被添加;在含有TAN-OL-RED和TAN-OHTy-RED的内脏杂碎的样品测试中,在4天后该值各自达到4000和5000MPN/100g,然后在8天后升高至12,000和14,000MPN/100g。
表3:碎牛肉的第一测试的结果。对照=未处理样品;样品A=用配方菜蓟属TAN 80/10/10处理的样品(具有可水解和浓缩的单宁);样品B=用配方TAN-OL-RED处理的样品,样品C=用TAN-OHTy-RED处理的样品。在制备后分析4天(细菌总数=TBC,UFC/g,参考ISO 2003/4833;pH参考CE 2005/2073mod.reg.CE 2007/1441)
实施例7
使用单宁提取物稳定动物饲料的样品
制备宠物食物(狗),配方基于谷类(56wt%),肉和肉产品(22wt%),油和脂肪(3wt%),完整大豆种子和亚麻籽(2wt%),蔬菜(1.5wt%),水,总计100。
将单独和/或与源自绿茶提取物,朝鲜蓟提取物,橄榄树叶,橄榄酱,葡萄籽提取物的抗氧化剂级分混合的0.3-0.7wt%的栗单宁提取物和其混合物添加进该对照配方中。
更具体地,在该实例中,在以0.7wt%的量添加TAN-OL-RED(实施例4的配方8)后测试该配方。
对照样品和用本发明的配方处理后的样品由饲养员评估这些化合物的适口性和处理后的动物的排泄物的质量。对于所有处理后的样品,总抗氧化剂和抗自由基能力通过在体外测试的稳定自由基DPPH方法来评估[对于在体外测试的稳定自由基,参见:Heimler D.,Vignolini P.,Dini M.G.,Vincieri F.F.,Romani A.“Antiradical activity and polyphenol composition ofBrassicaceae edible varieties”Food Chemistry,2006,99,464-469]。
使用本发明的配方处理的样品总是在营养质量和动物部分的反应性上具有更好的表现。
Claims (10)
1.单独或与其他多酚混合的栗单宁提取物和/或其级分作为抗氧化剂、抗微生物添加剂的用途和在食品原料和人类和动物食品中降低亚硝胺和真菌毒素浓度的用途。
2.根据权利要求1的用途,其中该栗单宁提取物及其级分在水中提取并且通过物理方法浓缩,不使用化学添加剂。
3.根据权利要求2的用途,其中所述物理方法优选由反渗透和纳滤表示。
4.根据前述权利要求的一项或多项的用途,其中将该单独或与其他多酚混合的栗单宁提取物及其级分以液体或固体状态应用至食品原料中或添加进食品的制备中。
5.根据前述权利要求的一项或多项的用途,其中该栗单宁提取物及其级分以与其他多酚混合的形式使用,所述多酚选自绿茶提取物,朝鲜蓟提取物,橄榄树叶,橄榄酱提取物,葡萄籽提取物,迷迭香提取物,甜菜根提取物。
6.根据前述权利要求的一项或多项的用途,其中该栗单宁提取物及其级分以液体的形式使用,其中相对于提取物或其级分的总重量,单宁的含量为1-40wt%,优选13-25wt%,或以粉末的形式使用,其中相对于提取物或其级分的总重量,单宁的含量为30-90wt%。
7.根据前述权利要求的一项或多项的用途,其中该栗单宁提取物和/或其级分,可能与其他多酚混合,通过种子处理的第一处理和谷穗的喷雾或撒粉的第二处理来应用,优选地第一处理相对于要处理的种子的总重量以0.1-10wt%浓度的粉末形式的单宁提取物来进行,其中所述粉末形式的单宁提取物的单宁含量为30-90wt%,并且第二处理相对于进行谷穗喷雾处理所使用的水或液体载体的总重量以0.5-5wt%浓度的液体形式的栗单宁提取物和/或其一种级分来进行,其中所述液体形式的栗单宁提取物和/或其一种级分的单宁含量为13-25wt%。
8.根据前述权利要求1-6中一项或多项的用途,其中该栗单宁提取物及其级分与甜菜根提取物一起被用作用于肉类防腐的添加剂。
9.根据前述权利要求1-6中一项或多项的用途,其中该栗单宁提取物及其级分,可能与其他多酚混合,被用作动物饲料添加剂,所述多酚来自绿茶提取物,朝鲜蓟提取物,橄榄树叶,橄榄酱提取物,葡萄籽提取物,迷迭香提取物,甜菜根提取物。
10.处理食品原料和人类和动物食品的方法,其包括根据前述权利要求1-9中一项或多项的单独和/或与其他多酚混合的栗单宁提取物和/或其级分的用途。
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