CN113875987A - 一种具有抗衰老功能的组方食品 - Google Patents
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Abstract
本发明公开了一种抗衰老增强免疫力的组方食品,涉及生物医药及食品领域,将副干酪乳杆菌、长双歧杆菌、约氏乳杆菌和植物乳杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中增殖发酵获得,其中灵芝浓度为16.6~66.4 mg/mL,当归浓度为15.5~60.0 mg/mL;添加益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌=2:1:1:1,副干酪乳杆菌在添加灵芝和当归的MRS肉汤培养基中初始CFU/mL=1~2*106,进行增殖并发酵至发酵液中总益生菌CFU/mL=4~5*109。利用益生菌发酵药食同源原料并制备成发酵液制剂,其抗衰老以及清除自由基的效果要优于药食同源原料或益生菌培养基溶液,产品的总抗氧化能力比较显著,具有优异的抗衰老并提高***免疫力的功效。
Description
技术领域
本发明涉及生物医药及食品领域,具体地,涉及一种具有抗衰老功能的组方食品。
背景技术
益生菌是指能够改变宿主某一部位菌群组成的且对宿主有益的活性微生物, 益生菌在人体健康中扮演非常重要的角色。目前, 益生菌已经被广泛的应用于医药、食品、护肤品等功能健康产品的制备。
药食同源是指即是食品也是药品的物质。中医学自古以来就有“药食同源”(又称为“医食同源”)理论。这一理论认为: 许多食物既是食物也是药物, 食物和药物一样同样能够防治疾病。截至2021年, 国家卫健委已经公布了近200种的药食同源物质名单。
衰老(Aging or Senescence)指随着年龄增加、机体逐渐出现的退行性变化。在生命代谢活动的过程中,会产生不同程度的自由基,它们可以单独存在,大多具有极强的氧化能力。有研究表明,自由基过多产生就会对人体组织造成伤害,从而导致机体加速衰老和发生各种疾病。
我们在前期研究中发现,将药食同源原料利用益生菌发酵后,制备得到的药食同源益生菌发酵液制剂,具有优异的抗衰老以及清除自由基的效果,其抗衰老以及清除自由基的效果要优于1)药食同源原料 、2) 益生菌培养基溶液以及3)清除自由基物质维生素C。目前,现有技术中未见利用益生菌组合发酵药食同源原料并获得发酵液制剂用于抗衰老产品的相关报道, 因此具有极佳的应用前景。
发明内容
有鉴于此,本发明提供了一种基于药食同源益生菌发酵液制剂的抗衰老功能组方食品及其制备方法,解决上述技术问题。
本发明采用的技术手段如下: 一种抗衰老增强免疫力的组方食品,其特征在于:将副干酪乳杆菌、长双歧杆菌、约氏乳杆菌和植物乳杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中增殖发酵获得,其中灵芝浓度为16.6~66.4 mg/mL,当归浓度为15.5~60.0mg/mL;添加益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌= 2:1:1:1, 副干酪乳杆菌在添加灵芝和当归的MRS肉汤培养基中初始CFU/mL=1~2*106,进行增殖并发酵至发酵液中总益生菌CFU/mL=4~5*109。
进一步地,培养发酵结束后将药食同源益生菌的发酵液离心, 取上清液,再将上清液高温喷雾干燥后制得粉末,包装入小袋,制得抗衰老功能的复合型食品。
进一步地,灵芝和当归水提液分别是用药食同源的灵芝和当归超细粉经过熬煮制成的。药食同源的灵芝和当归超细粉的制备方法是: 将食用级别的灵芝、当归分别在温度50~60℃、压力-0.008~-0.01MPa的条件下,保持5~8小时进行干燥,再经冷却后进行粉碎,使粉碎粒度为800~1200目,即分别得到超微灵芝粉、超微当归粉。
进一步地,含有灵芝和当归水提液的MRS肉汤培养基是将灵芝和当归水提液加入MRS肉汤培养基后,32℃下培养24h。
进一步地,先将副干酪乳杆菌和长双歧杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中,37℃培养12h后,再加入约氏乳杆菌和植物乳杆菌,再进行增殖发酵。
本发明是一种基于药食同源益生菌发酵液制剂的抗衰老功能组方食品及其制备方法,与现有技术和产品相比, 本发明具有如下的优点:
(1) 本发明将药食同源原料利用益生菌发酵后,制备得到的药食同源的益生菌发酵液制剂,对衰老机制有显著的调控作用, 其抗衰老以及清除自由基的效果要优于药食同源原料或益生菌培养基溶液。
(2) 本发明的抗衰老组方食品对氮基自由基(DPPH)的清除能力、对羟基自由基的清除能力, 对提高总抗氧化能力(T-AOC)水平有显著作用,具有优异的抗衰老并提高***免疫力的功效。
(3) 本发明的抗衰老组方食品生物安全性很高。
(4) 本发明的抗衰老组方食品的口感很好、因此本产品的接受度很好。
具体实施方式
为便于理解本发明,本发明列举实施例如下。本领域技术人员应该明了,所述实施例仅仅是帮助理解本发明,不应视为对本发明的具体限制。
清除氮基自由基(DPPH)能力的测试
DPPH自由基一种稳定的氮中心自由基, 是测量样本抗氧化能力的重要指标之一,被广泛应用于抗衰老类食品、保健品及药品的研究中。根据氮基自由基测定试剂盒(北京索莱宝生物科技有限公司)说明书步骤进行操作。称取DPPH粉末制得0.2mmol/L的DPPH无水乙醇混合溶液。准确吸取200μL无水乙醇溶液和200μL样品待测溶液, 震荡混匀; 200μL蒸馏水和200μL DPPH无水乙醇混合液, 震荡混匀。于酶标仪中测得待测溶液的吸光度A517nm。
清除率(%)=[1-(As-Ab)/Ac]×100% (1)
式(1)中: As为样品溶液和DPPH无水乙醇混合液各200μL的OD值; Ab为无水乙醇和样品溶液各200μL的OD值; Ac为DPPH无水乙醇混合液和蒸馏水各200μL的OD值。
清除羟基自由基能力的测试
羟自由基(·OH)导致体内代谢紊乱引起疾病并导致衰老。羟自由基清除能力是样品抗氧化能力的重要指标之一, 在抗衰老类保健品和药品研究中得到广泛应用。根据羟基自由基测定试剂盒(北京索莱宝生物科技有限公司)说明书步骤进行操作, 利用酶标仪测得其吸光度A550nm。蒸馏水作空白对照, 维生素C作阳性对照药物。
清除率(%)=(Ac-As)/(Ac-Ab)×100% (2)
式中:Ac为对照管的OD值;As为测定管的OD值;Ab为空白管的OD值。
总抗氧化能力的测定
总抗氧化能力(T-AOC assay)测定试剂盒是一种采用ABTS作为显色剂,可以对各种溶液的总抗氧化能力进行快速检测的试剂盒。根据总抗氧能力测定试剂盒(北京索莱宝生物科技有限公司)说明书步骤进行操作,于酶标仪下测得各个样品的吸收度A405nm,实验中蒸馏水为空白对照, 维生素C作阳性对照。
总抗氧化能力=A0-Ax (3)
公式(3)中:A0为测定管的OD值; Ax为对照管的OD值。所有数据均为3次平行测定的平均值,表示为平均值±标准差(Mean±SD)。采用Prism8.0软件对试验结果进行统计分析,Duncan多重比较进行单因素方差分析,P<0 .05代表差异显著。
实施例1
一种抗衰老增强免疫力的组方食品,制备方法是:
将食用级别的灵芝、当归分别在温度50~60℃、压力-0.008~-0.01MPa的条件下,保持5~8小时进行干燥,再经冷却后进行粉碎,使粉碎粒度为800~1200目,分别得到超微灵芝粉、超微当归粉。
将上述药食同源的灵芝和当归超细粉分别熬煮制成灵芝和当归水提液。
将灵芝和当归水提液加入MRS肉汤培养基中,其中灵芝浓度为16.6 mg/mL,当归浓度为60.0 mg/mL,32℃下培养24h,然后将副干酪乳杆菌和长双歧杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中,MRS肉汤培养基中副干酪乳杆菌的初始CFU/mL=2*106,37℃培养12h后,再分别加入氏乳杆菌和植物乳杆菌,添加益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌= 2:1:1:1,再进行增殖发酵至发酵液中总益生菌CFU/mL=4.0*109。
实施例2
参考实施例1,将灵芝和当归水提液加入MRS肉汤培养基中,其中灵芝浓度为30mg/mL,当归浓度为30 mg/mL,32℃下培养24h,然后将副干酪乳杆菌和长双歧杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中,MRS肉汤培养基中副干酪乳杆菌的初始CFU/mL=1.5*106,37℃培养12h后,再分别加入氏乳杆菌和植物乳杆菌,添加益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌= 2:1:1:1,再进行增殖发酵至发酵液中总CFU/mL=5*109。
培养发酵结束后将药食同源益生菌的发酵液离心, 取上清液,再将上清液高温喷雾干燥后制得粉末,包装入小袋,制得抗衰老功能的复合型食品。
实施例3
参考实施例1,参考实施例1,将灵芝和当归水提液加入MRS肉汤培养基中,其中灵芝浓度为66.4 mg/mL,当归浓度为15.5 mg/mL,32℃下培养24h,然后将副干酪乳杆菌和长双歧杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中,MRS肉汤培养基中副干酪乳杆菌的初始CFU/mL=1*106,37℃培养12h后,再分别加入氏乳杆菌和植物乳杆菌,添加益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌= 2:1:1:1,再进行增殖发酵至发酵液中总CFU/mL=4.5*109。
对比例1:药食同源(灵芝、当归)的水提物溶液
药食同源的水提物溶液的制备步骤如下: 将灵芝细粉和当归细粉以1:1的质量配比混合, 并与30%的酒精溶液配置成10mg/ml的浑浊液。将溶液加热至沸腾, 在萃取装置中回流萃取4小时后,滤掉灵芝细粉和当归细粉的残渣, 制备成水提物溶液。
对比例2:益生菌的培养基溶液
将副干酪乳杆菌、长双歧杆菌、约氏乳杆菌、植物乳杆菌在MRS肉汤培养基中混合培养,MRS肉汤培养基中副干酪乳杆菌的初始CFU/mL=2*106,益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌= 2:1:1:1,每12 h在MRS琼脂培养基上进行活菌计数,第12h~36h为稳定期并调整混合菌液总CFU/ mL为4.0*109。
对比例3,维生素C
实施例与对比例的抗氧化活性测试结果如下:
从以上实验结果可见,利用益生菌组合发酵药食同源并制备成发酵液制剂,其抗衰老以及清除自由基的效果要优于药食同源原料、优于益生菌培养基溶液,并优于传统的清除自由基物质维生素C。表明利用益生菌组合发酵药食同源并制备成发酵液制剂后, 其清除自由基的能力得到了很大的增强, 非常适合用于抗衰老组方产品的制备。
实施例4
取上述实施例2的抗衰老食品进行临床试吃实验。在中国医科大学附属盛京医院体检科和保健科选取年龄在35~55岁的易衰老、免疫力低下的志愿者20例, 取得试吃实验同意书,分为两个组,每组10例。第一组口服上述实施例的抗衰老食品,每次口服5g(1汤匙、早中晚各一次),连服30日,观察并记录结果; 第二组为对照组,口服普通螺旋藻片剂、葡萄籽精华胶囊、汤臣倍健蛋白粉或含有三七总皂苷的保健食品中的任意一种, 每次口服5g(每次一片或一个胶囊、早中晚各一次), 连服30日,观察并记录结果。
其中,对所述“活化”以及“发酵培养”,本领域技术人员根据发明内容及现有技术应当容易理解和确认,本发明中一般选用。
申请人声明,本发明通过上述实施例来说明本发明的详细工艺设备和工艺流程但本发明并不局限于上述详细工艺设备和工艺流程,即不意味着本发明必须依赖上述详细工艺设备和工艺流程才能实施。所属技术领域的技术人员应该明了,对本发明的任何改进,对本发明产品各原料的等效替换及辅助成分的添加、具体方式的选择等,均落在本发明的保护范围和公开范围之内。
Claims (6)
1.一种抗衰老增强免疫力的组方食品,其特征在于:将副干酪乳杆菌、长双歧杆菌、约氏乳杆菌和植物乳杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中增殖发酵获得,其中灵芝浓度为16.6~66.4 mg/mL,当归浓度为15.5~60.0 mg/mL;添加益生菌菌量比为副干酪乳杆菌:长双歧杆菌:约氏乳杆菌:植物乳杆菌= 2:1:1:1, 副干酪乳杆菌在添加灵芝和当归的MRS肉汤培养基中初始CFU/mL=1~2*106,进行增殖并发酵至发酵液中总益生菌CFU/mL=4~5*109。
2.根据权利要求1所述的一种抗衰老增强免疫力的组方食品,其特征在于:培养发酵结束后将药食同源益生菌的发酵液离心, 取上清液,再将上清液高温喷雾干燥后制得粉末,包装入小袋,制得抗衰老功能的复合型食品。
3.根据权利要求1所述的一种抗衰老增强免疫力的组方食品,其特征在于:灵芝和当归水提液分别是用药食同源的灵芝和当归超细粉经过熬煮制成的。
4.根据权利要求3所述的一种抗衰老增强免疫力的组方食品,其特征在于:药食同源的灵芝和当归超细粉的制备方法是: 将食用级别的灵芝、当归分别在温度50~60℃、压力-0.008~-0.01MPa的条件下,保持5~8小时进行干燥,再经冷却后进行粉碎,使粉碎粒度为800~1200目,即分别得到超微灵芝粉、超微当归粉。
5.根据权利要求1所述的一种抗衰老增强免疫力的组方食品,其特征在于:含有灵芝和当归水提液的MRS肉汤培养基是将灵芝和当归水提液加入MRS肉汤培养基后,32℃下培养24h。
6.根据权利要求1所述的一种抗衰老增强免疫力的组方食品,其特征在于:先将副干酪乳杆菌和长双歧杆菌添加到含有灵芝和当归水提液的MRS肉汤培养基中,37℃培养12h后,再加入约氏乳杆菌和植物乳杆菌,再进行增殖发酵。
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