CN113230303A - 九龙藤提取物及其制备方法以及在食源性致病菌抗菌剂中的应用 - Google Patents
九龙藤提取物及其制备方法以及在食源性致病菌抗菌剂中的应用 Download PDFInfo
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Abstract
本发明提供了一种九龙藤提取物,该提取物中,以黄苓苷计,总黄酮含量为45‑55%。九龙藤提取物的制备方法,包括以下步骤:(1)称取干燥的九龙藤研磨后过筛,得到九龙藤粉末;(2)将步骤(1)得到的九龙藤粉末,用有机溶媒结合超声辅助提取得到九龙藤提取物溶液;(3)将步骤(2)得到的九龙藤提取物溶液,经分离和浓缩后得到九龙藤提取物浓缩液;(4)将步骤(3)得到九龙藤提取物浓缩液冷冻干燥后得到九龙藤提取物粉末。本发明的九龙藤提取物可用于预防由常见的食源性致病菌引起的食源性疾病,该提取物对多种食源性致病菌的抑菌效果明显,可作为天然的具有广泛抗菌用途的抗菌剂。因此,它可以作为一种广谱抑菌剂应用于食品和动物饲料行业中。
Description
技术领域
本发明涉及食品安全技术领域,具体地,涉及九龙藤提取物及其制备方法以及在食源性致病菌抗菌剂中的应用。
背景技术
食以安为本,食品安全与民生紧密相关,微生物引起的食品污染是我国食品安全的主要问题,而细菌污染则是微生物污染的主体。根据WHO的统计,全球由食源性致病菌引起的患者中,70%是由不同致病性微生物污染食物和饮水造成。近年来,全球食源性疾病的病发率逐年升高,造成的危害日益加重,已成为威胁食品安全的最大问题。常见的食源性致病菌主要包括大肠埃希氏菌,沙门氏菌,志贺氏菌,单增李斯特菌,副溶血性弧菌,溶血性链球菌和金黄色葡萄球菌。在食品加工和储存过程中,这些食源性致病菌会诱发食品腐败,可引起食物中毒或引起食源性疾病。传统的抗菌剂已不能满足人们的生活需求。目前市场上的抗菌剂在实际使用过程中存在抑菌效果不理想、抑菌持久性差、抑菌剂损失严重等问题,急需研发绿色无污染、成本低、抑菌效果好并且抑菌效果持久的产品。针对于无毒的新型抗菌剂的迫切需求,越来越多的目光投入到具有天然活性的抗菌物质研究中。从天然物质中筛选出具有广谱抗氧化、抗菌活性的天然食品添加剂用于降低或清除食品中的有害微生物,保证食品质量,延长食品保质期,具有重要意义。
目前我国对于食源性疾病的防治主要以抗生素等化学药物为主,但由于长期滥用现已造成环境污染和病菌抗药等一系列问题。与化学药物相比,中草药具有“三效”(高效、速效、长效)、“三小”(剂量小、毒性小、副作用小)、不易产生抗药性、残留少等特点,因此植物源杀菌药物可用作化学药物的有效替代品。《中药大词典》中收录了我国传统中草药有4773种,其药效和安全性都较为明确,具有抗菌防腐作用的植物有2000多种,并且大量研究发现,丁香、迷迭香、肉桂、黄芩、绿茶、五味子、花椒、牛至等中草药及香辛料的提取物均具有不同程度的抑菌作用。我国植物资源丰富,不少植物提取物中含有抑制病原微生物、抗菌、抗炎、抗癌等生物活性成分,且无毒、无害、无残留,食用安全,其中从人们食用的中草药和香辛料中提取天然抗菌剂被认为是食品添加剂研究领域最有前景的发展方向。
九龙藤(Bauhinia championii)别名过岗龙、过江龙等,是豆科苏木亚科羊蹄甲属有花植物。分布于越南、印度尼西亚、印度以及中国的广东、湖南、湖北、江西、福建、浙江、贵州、广西等地,生长于海拔200-1000米的地区,一般生于丘陵灌丛、山地疏林以及密林中,尚未人工引种栽培。福建福州永泰县青云山地区九龙藤资源丰富。九龙藤性平、苦涩,无毒,主要含有黄酮类、多糖、挥发油等,具有抗炎镇痛、消除自由基、抗菌、抗肿瘤等功效,在畲医中作为一种治疗痹症的特效药,常用于治疗风湿疼痛。九龙藤在我国植物资源丰富,其药材基源为根或茎,九龙藤根含黄酮,龙藤甙,没食子酸等成分鞣质含量高达20.76%。尽管九龙藤是一种重要的中药材,在医学领域有着广泛的用途,但是目前尚未有研究发现其对食源性致病菌的抗菌性能。
发明内容
针对现有技术中的缺陷,本发明的目的是提供一种九龙藤提取物及其制备方法以及在食源性致病菌抗菌剂中的应用。
本发明的目的是通过以下方案实现的:
本发明的第一方面提供一种九龙藤提取物,该提取物中,以黄苓苷计,总黄酮含量为45-55%。
本发明的第二方面提供一种九龙藤提取物的制备方法,包括以下步骤:
(1)称取干燥的九龙藤研磨后过筛,得到九龙藤粉末;九龙藤植物包括广东、广西、福建、中国台湾、浙江等地九龙藤,材料优选为九龙藤的根部和茎部;研磨后优选过20目筛;
(2)将步骤(1)得到的九龙藤粉末,用有机溶媒结合超声辅助提取得到九龙藤提取物溶液;
(3)将步骤(2)得到的九龙藤提取物溶液,经分离和浓缩后得到九龙藤提取物浓缩液;
(4)将步骤(3)得到九龙藤提取物浓缩液置于-80℃冰箱冷冻过夜后,经真空冷冻干燥机干燥后得到九龙藤提取物粉末呈深褐色,易溶于有机溶剂和水。优选地,所述真空冷冻干燥的条件为:-50℃,4小时;-35℃,10小时;-10℃,10小时;25℃,24小时。
优选地,步骤(2)中,所述有机溶媒为80%(v/v)乙醇,九龙藤提取物粉末与乙醇的添加比例为1:10-1:20,超声提取的参数为40℃,480W下进行一小时。
优选地,步骤(3)中,分离方法为:900-1000g离心10-15分钟,收集上层清液。
优选地,浓缩方法为使用旋转蒸发仪蒸发浓缩上层清液,浓缩比例为原体积的四分之一到五分之一。
本发明提供的九龙藤提取物经真空冷冻干燥后,为深褐色粉末,易溶于水,乙醇,DMSO等溶剂。因此,本发明提供的九龙藤提取物本身就是一个具有很好的成药性能的植物药原料。
本发明的第三方面提供九龙藤提取物在食源性致病菌抗菌剂中的应用。
优选地,所述九龙藤提取物能抑制和杀灭革兰氏阴性菌和革兰氏阳性菌。
优选地,所述食源性致病菌包括金黄色葡萄球菌(Staphylococcus aureus),单核细胞增生李斯特菌(Listeria monocytogenes),大肠杆菌(Escherichia coli),肠炎沙门氏菌(Salmonella enterica Enteritidis)。
优选地,所述九龙藤提取物对金黄色葡萄球菌和李斯特菌的最小抑菌浓度分别为0.39mg/mL和1.56mg/mL,所述九龙藤提取物对大肠杆菌和肠炎沙门氏菌的最小抑菌浓度分别为6.25mg/mL和12.5mg/mL。
优选地,所述应用包括,将所述九龙藤提取物作为广谱抑菌剂应用于食品和动物饲料中。
与现有技术相比,本发明具有如下的有益效果:
本发明通过使用超声波辅助提取方法,提高了九龙藤中抗菌化合物的提取效率和得率,实现在短时间内获得大量的九龙藤提取物。提取出的九龙藤抗菌成分,对造成食物中毒及食源性疾病的细菌(金黄色葡萄球菌,单增李斯特菌,大肠杆菌,肠炎沙门氏菌)具有优越的广谱抗菌活性,而且在低浓度下也能很好地发挥作用,为开发出抗菌性更强,抗菌范围更广的天然抗菌剂奠定了基础。
附图说明
通过阅读参照以下附图对非限制性实施例所作的详细描述,本发明的其它特征、目的和优点将会变得更明显:
图1为不同浓度的九龙腾提取物对于四种常见食源性致病菌的抗菌活性效果图;
图2为不同浓度的九龙腾提取物对于四种常见食源性致病菌的抑菌圈直径柱状对比图。
具体实施方式
下面结合具体实施例对本发明进行详细说明。以下实施例将有助于本领域的技术人员进一步理解本发明,但不以任何形式限制本发明。应当指出的是,对本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变化和改进。这些都属于本发明的保护范围。
本发明在研究中草药提取物的抗菌活性,以获得抗菌能力和抗菌范围更优的抗菌剂时,发现国内植物资源丰富的中草药九龙藤对造成食物中毒及食源性疾病的细菌(金黄色葡萄球菌,单增李斯特菌,大肠杆菌,肠炎沙门氏菌)具有优越的广谱抗菌活性,而且在低浓度下也能很好地发挥作用,可用于预防由上述常见致病菌引起的食源性疾病。九龙藤提取物对多种食源性致病菌的抑菌效果明显,可作为天然的具有广泛抗菌用途的抗菌剂。因此,它可以作为一种广谱抑菌剂应用于食品和动物饲料行业中。
实施例1.九龙腾提取物的制备
将干燥的九龙藤粉碎后过20-40目筛,取4g九龙腾粉末加入40-80ml的80%(V/V)乙醇在40℃和480W下用超声辅助提取方法提取1-2小时,重复提取两次以提高九龙藤中抗菌化合物的提取效率和得率。九龙腾提取物溶液在室温(900-1000g,10-15分钟) 下离心,收集上层清液。获得的上层清液使用旋转蒸发仪在40℃下蒸发浓缩至原体积的四分之一。浓缩后的提取物置于-80℃冰箱冷冻过夜后,用真空冷冻干燥机干燥即可得到九龙藤提取物粉末,真空冷冻干燥的条件为:-50℃,4小时;-35℃,10小时;-10℃, 10小时;25℃,24小时。该提取物中总黄酮含量为45-55%(以黄苓苷计)。对比实验室前期只采用乙醇提取的方法,九龙藤提取物经乙醇辅助超声波提取后其中总黄酮的含量由原来的40%左右提升到了45%-55%,得率显著提升。
实施例2.九龙腾提取物的抗菌实验
将实施例得到的九龙藤提取物粉末以不同浓度(25mg/mL,50mg/mL,100mg/mL 和200mg/mL)溶于二甲基亚砜(DMSO)中,以进行抗菌活性研究。本发明选用四种常见的食源性致病菌:金黄色葡萄球菌(S.aureus ATCC 25923),单核细胞增生李斯特菌(L.monocytogenes ATCC 7644),大肠杆菌(E.coli ATCC 252922)和血清型肠炎沙门氏菌(S.enterica Enteritidis ATCC1307637)。首先将胰蛋白胨大豆肉汤(TSB) 琼脂板上生长的单个菌落接种到TSB培养基中,并在37℃和250rpm条件下振荡过夜培养。将细菌悬浮液调为1×106CFU/mL的浓度用于以下抗菌实验。本发明所用菌株,培养基及化学试剂皆为购买获得。
1、抑菌圈DIZ的测量。
为了测试九龙藤提取物对食源性致病菌的广谱抗菌作用,本发明设置了四个不同浓度的九龙腾提取物(25mg/mL,50mg/mL,100mg/mL和200mg/mL)处理上述四种食源性致病菌。根据临床实验室标准协会(CLSI)所述的琼脂扩散法评估九龙藤提取物的抗菌活性。
用无菌LB培养基将上述四种食源性致病菌稀释至约1×106CFU/mL,然后通过无菌玻璃珠(直径6mm)将100uL的每种细菌悬浮液均匀地分散在LB琼脂平板的表面上。将牛津杯(内径为6mm,外径为7.8mm,高度为10mm)轻轻放在琼脂表面上,然后向牛津杯中滴加60微升不同浓度的九龙藤提取物(25mg/mL,50mg/mL,100mg/mL和 200mg/mL),37℃培养16~18小时,测量抑菌圈的直径,实验重复两次,每次三个平行。使用氨苄青霉素(10ul/ml)作为阳性对照,DMSO(60uL/cup)作为阴性对照。九龙藤提取物的抗菌作用,可以用抑菌圈DIZ来评估,可以分为很强(DIZ≥20mm),强 (15mm≤DIZ<20mm),中等(10mm≤DIZ<15mm)以及弱(DIZ<10mm)。DIZ 值小于或等于8.0mm的被认为没有抗菌活性。
如图1和图2所示,结果表明:九龙藤提取物对上述四种常见食源性致病菌呈现出明显的抗菌作用。低浓度的九龙藤提取物(25mg/mL)对金黄色葡萄球菌(S.aureus) 即有很强的抗菌活性(对应DIZ 17.6mm);50mg/mL,100mg/mL和200mg/mL的九龙藤提取物对单增李斯特菌(L.monocytogenes)表现出优越的抗菌能力(对应DIZ 16.2 mm,16.8mm和18.7mm);100mg/mL和200mg/mL的九龙藤提取物对大肠杆菌(E.coli) 有很强的抗菌活性(对应DIZ14.9mm和15.2mm);同时100mg/mL和200mg/mL的九龙藤提取物对肠炎沙门氏菌(S.enterica)也有明显抑制的作用(对应DIZ 14.8mm 和15.8mm)。
2、确定最小抑菌浓度MIC和最小杀菌剂浓度MBC
MIC及MBC测定参照临床实验室标准化协会(CLSI)发布的肉汤微量稀释法的标准方案进行。通过连续96稀释微孔板法和水解酪蛋白(MH)琼脂计数法测定九龙藤提取物的MIC和MBC。
通过连续96稀释微孔板法和Mueller-Hinton(MH)琼脂计数法测定九龙藤提取物的MIC和MBC。
将100uL水解酪蛋白肉汤培养基添加到96孔培养板中,然后将100mg/mL的九龙藤提取物连续两次稀释,最终浓度范围为0.39mg/mL至50mg/mL。依次向各个孔中添加不同浓度的九龙藤提取物20uL。之后将100uL的标准细菌悬浮液(1X 106CFU/mL) 添加到每个测试孔中,以使每个孔中的最终体积为220uL,将孔板在37℃下孵育24小时。孵育后,将30uL新鲜制备的刃天青(质量浓度为0.015%)添加到所有测试孔中,并进一步孵育2小时,以使存活的微生物将蓝色刃天青染料代谢为粉红色刃天青。另设阳性对照(100微升106CFU/mL的菌液加水解酪蛋白肉汤培养基)、阴性对照(只含水解酪蛋白肉汤培养基)。生长48小时后以在小孔内完全抑制细菌生长的最低药物浓度为MIC值(小孔内颜色依然为蓝色)。在最低抑菌浓度及以上浓度的孔中取肉汤10微升,接种于水解酪蛋白平板上,将没有菌落生长的最低浓度定义为MBC。氨苄青霉素用作阳性对照(使用浓度分别为25ug/mL,12.5ug/mL,6.25ug/mL,3.125ug/mL,1.56ug/mL, 0.78ug/mL,0.39ug/mL,0.2ug/mL)实验重复三次,每次三个平行。。
表1九龙藤提取物对四种食源性致病菌的MIC和MBC值
结果表明:九龙藤提取物对四种食源性沙门氏菌的MIC和MBC值也证明九龙藤提取物具有高效广谱的抗菌活性。针对革兰氏阳性细菌,如金黄色葡萄球菌(S.aureus) 和李斯特菌(L.monocytogenes),九龙藤提取物的MIC值很低,分别为0.39mg/mL和 1.56mg/mL,表现出良好的抗菌活性。针对革兰氏阴性细菌,如大肠杆菌(E.coli) 和肠炎链球菌(S.enterica Enteritidis),九龙藤提取物的MIC值分别为6.25mg/mL 和12.5mg/mL。
对比例1
参见专利文献CN111265446A公开的一种鱼腥草提取物,该提取物经过分离纯化后其中的总黄酮含量达到80%以上,100mg/ml的鱼腥草提取物对大肠杆菌的抑菌直径达10mm;对金黄色葡萄球菌的抑菌直径达15mm。本发明公开的九龙藤提取物在未经分离纯化的情况下,100mg/ml九龙藤提取物对大肠杆菌的抑菌圈直径可达14.9mm;对金黄色葡萄球菌的抑菌直径可达20mm,其抑菌效果明显优于该鱼腥草提取物。
对比例2
参见专利文献CN106889173A公开的一种菱角壳黄酮粗提取物和纯化物,其对金黄色葡萄球菌的最低抑菌浓度分别为12.5mg/ml和3.125mg/ml,本发明中的九龙藤提取物对金黄色葡萄球菌的最低抑菌浓度显著低于菱角壳黄酮粗提取物和纯化物,证明其具有优越的抑菌活性。
由于革兰氏阳性菌和革兰氏阴性菌的细胞壁结构不同,中草药提取物往往对革兰氏阳性菌具有明显的抑菌效果,很多文献的研究也是集中于此,对于中草药提取物抑制革兰氏阴性菌生长的研究很少。本发明中的九龙藤提取物不仅对革兰氏阴性菌具有显著的抑菌活性,对革兰氏阴性菌的抑菌活性也很明显,证明本发明中的九龙藤提取物具有广谱高效的抑菌活性。
综上所述,本发明通过超声波辅助方法高效快速的提取九龙藤中的抗菌化合物,并通过体外抑菌试验发现低浓度的九龙藤提取物就表现出强大的抗菌作用。这说明九龙藤是开发广谱高效抗菌剂的一个新来源,可用于由食源性致病菌引起的食物中毒和食源性疾病,具有极大的推广价值。
以上对本发明的具体实施例进行了描述。需要理解的是,本发明并不局限于上述特定实施方式,本领域技术人员可以在权利要求的范围内做出各种变化或修改,这并不影响本发明的实质内容。在不冲突的情况下,本申请的实施例和实施例中的特征可以任意相互组合。
Claims (10)
1.一种九龙藤提取物,其特征在于,该提取物中,以黄苓苷计,总黄酮含量为45-55%。
2.一种九龙藤提取物的制备方法,其特征在于,包括以下步骤:
(1)称取干燥的九龙藤研磨后过筛,得到九龙藤粉末;
(2)将步骤(1)得到的九龙藤粉末,用有机溶媒结合超声辅助提取得到九龙藤提取物溶液;
(3)将步骤(2)得到的九龙藤提取物溶液,经分离和浓缩后得到九龙藤提取物浓缩液;
(4)将步骤(3)得到九龙藤提取物浓缩液冷冻干燥后得到九龙藤提取物粉末。
3.根据权利要求2所述的九龙藤提取物的制备方法,其特征在于,步骤(2)中,所述有机溶媒为80%(v/v)乙醇,添加比例为1:10-1:20。
4.根据权利要求2所述的九龙藤提取物的制备方法,其特征在于,步骤(3)中,分离方法为:900-1000g离心10-15分钟,收集上层清液。
5.根据权利要求2所述的九龙藤提取物的制备方法,其特征在于,浓缩方法为使用旋转蒸发仪蒸发浓缩上层清液,浓缩比例为4:1-5:1。
6.如权利要求1所述的九龙藤提取物在食源性致病菌抗菌剂中的应用。
7.根据权利要求6所述的九龙藤提取物在食源性致病菌抗菌剂中的应用,其特征在于,所述九龙藤提取物能抑制和杀灭革兰氏阴性菌和革兰氏阳性菌。
8.根据权利要求6或7所述的九龙藤提取物在食源性致病菌抗菌剂中的应用,其特征在于,所述食源性致病菌包括金黄色葡萄球菌,单核细胞增生李斯特菌,大肠杆菌,肠炎沙门氏菌。
9.根据权利要求8所述的九龙藤提取物在食源性致病菌抗菌剂中的应用,其特征在于,所述九龙藤提取物对金黄色葡萄球菌和李斯特菌的最小抑菌浓度分别为0.39mg/mL和1.56mg/mL,所述九龙藤提取物对大肠杆菌和肠炎沙门氏菌的最小抑菌浓度分别为6.25mg/mL和12.5mg/mL。
10.根据权利要求6所述的九龙藤提取物在食源性致病菌抗菌剂中的应用,其特征在于,用所述九龙藤提取物作为广谱抑菌剂应用于食品和动物饲料中。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2316465A1 (en) * | 2009-10-15 | 2011-05-04 | Centre National De La Recherche Scientifique | Use of extract of leeches' saliva as anti-bacterial agent for the manufacture of different compositons |
CN107573221A (zh) * | 2017-08-23 | 2018-01-12 | 长江大学 | 一种龙须藤提取物及其制备方法与应用 |
CN112294885A (zh) * | 2020-10-08 | 2021-02-02 | 湖南金凤凰中草药有限责任公司 | 一种快速散结消肿,化瘀生机,止痒止痛的抑菌液中药及配制方法 |
-
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2316465A1 (en) * | 2009-10-15 | 2011-05-04 | Centre National De La Recherche Scientifique | Use of extract of leeches' saliva as anti-bacterial agent for the manufacture of different compositons |
CN107573221A (zh) * | 2017-08-23 | 2018-01-12 | 长江大学 | 一种龙须藤提取物及其制备方法与应用 |
CN112294885A (zh) * | 2020-10-08 | 2021-02-02 | 湖南金凤凰中草药有限责任公司 | 一种快速散结消肿,化瘀生机,止痒止痛的抑菌液中药及配制方法 |
Non-Patent Citations (2)
Title |
---|
张玉琴,等: "龙须藤研究进展", 《亚太传统医药》 * |
黄琼,等: "超声波辅助提取龙须藤总黄酮及其抗氧化活性的研究", 《食品科技》 * |
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