CN116058494A - 一种补气固发的益生菌发酵药食同源饮料及其制备方法 - Google Patents
一种补气固发的益生菌发酵药食同源饮料及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种具有补气固发的益生菌发酵药食同源饮料及其制备方法,所述药食同源的比例为:栀子3份,百合6份,黄芪1份,党参1份;益生菌发酵的菌种为:鼠李糖乳杆菌、植物乳杆菌、格式乳杆菌、罗伊氏乳杆菌、长双歧杆菌、发酵乳杆菌、乳酸乳球菌、嗜酸乳杆菌、保加利亚乳杆菌中的一种或多种。益生菌菌种经甘蔗糖蜜培养基活化后接种至灭菌药食同源基质中,液态发酵24小时后滤渣即获得一种补气固发的益生菌发酵药食同源饮料。该饮料通过急毒试验验证无任何毒副作用,通过脱发小鼠诱导重新生发,说明本发明物安全可食用,且具有诱导重新生发,改善发质的作用。
Description
技术领域
本发明涉及一种食品,具体涉及一种补气固发的乳酸菌发酵药食同源饮料及其制备方法,属于功能饮料制备和应用领域。
背景技术
脱发是困扰中年人的生活难题之一,人群庞大,市场上缓解脱发的产品种类繁多,以米诺地尔类OTC产品为龙头,其次为药食同源洗发露,种类繁多,市场竞争激烈。这部分兴起的产品大多为外用产品,使用后效果反复,易反弹,不能从根本上解决脱发问题,最新有研究指出肠道菌群与脱发有着密切关系,如Cell Reports指出小鼠肠道内菌群合成生物素可有效改善发质,防止头发断裂,促进脱发小鼠重新生长出毛发。所以调整肠道菌群平衡,提高体内生物素含量能够在治疗脱发的过程中发挥作用。
中医学认为脱发的原因有两点:一是血热风燥说,《儒门事亲》说:“年少早白落,此血热太过也。世俗只知发者血之余,血衰故耳。岂知血热而发反不茂肝者,木也。火多水少,木反不荣;火至于顶,炎上之甚也。热病汗后,发多脱落,岂有寒耶。”血为水谷精微所化,以奉养周身。血热生风,风热随气上窜于巅顶,毛根得不到阴血的滋养,头发则会突然脱落或焦黄或早白等。二是脾胃湿热说,《临证指南》说:“湿从内生者,必其人膏粱酒鳢过度。”说明恣食肥甘,容易伤胃损脾,湿热内蕴,发根循经上蒸巅顶,侵蚀发根白浆,导致头发粘腻,头发稀少或者均匀性脱发。
百合性甘、微寒,归肺、心经;有润肺养阴、失眠多梦、热病后的余热未清、虚烦不安等功效。栀子性苦、寒,归心、肝、肺、胃、三焦经;有泻火除烦、凉血止血解毒、清利湿热、肝经湿热郁火等功效。党参性甘、平,归脾、肺经;有补中益气、养血补津等功效。黄芪性甘、微温,归脾、肺经;有补气健脾升阳、益卫固表止汗、利尿,托疮生肌等功效。生物素有促进汗腺、神经组织、骨髓、男性性腺、皮肤及毛发的正常运作和生长、减轻湿疹、皮炎症状、预防白发及脱发、有助于治疗秃顶等功效。
发明内容
本发明的目的是提供一种乳酸菌发酵药食同源饮料,以改善现在人群气亏导致严重脱发的技术问题。
本发明主要是通过以下技术方案解决上述技术问题的:一种益生菌发酵药食同源饮料,包括乳酸菌菌种、栀子、百合、黄芪、党参,所述益生菌菌种为高产生物素乳酸菌,栀子等药食同源为中医治疗脱发的成分。
一种补气固发的乳酸菌发酵药食同源组合物的饮料及其制备方法,包括以下步骤:
步骤A:发酵种子液制备
将高产生物素菌株分别按2%分别接种于甘蔗糖蜜培养基中,置于37℃静置培养24小时,获得发酵种子液。
步骤B:发酵基质制备
将药食同源组合物转移至发酵罐中,加入纯化水后使用纯蒸汽进行115℃,30分钟杀菌,冷却至常温后获得发酵基质。
步骤C:将步骤1制备发酵种子液按比例接种至步骤2制备发酵基质中,恒温搅拌发酵,发酵产物为一种补气固发益生菌发酵药食同源组合物饮料。
本发明具有以下优点:
1. 本发明口味独特、食用健康、有效预防脱发;
2. 发酵菌种能够高产生物素,药食同源组合物具有祛火生血的功能;
3. 发酵后活菌含量高达108cfu/mL。
附图说明
附图用来提供对本发明的进一步理解,并且构成说明书的一部分,与本发明的实施一起用于解释本发明,并不构成对本发明的限制。
附图1为脱发小鼠服用本发明物后诱导重新生发的机制。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。
本案例的实施物料,检测方法,均符合国家相关标准要求。
实施例1:筛选高产生物素菌株
恒康生物微生物菌种保藏中心(HKMCC)现保存81株乳酸菌收录于中国可用于食品的菌种名单。从此名单中初步筛选抗逆性强的12株乳酸菌进一步进行高产生物素菌株的筛选工作,具体如下:
步骤A:强抗逆性菌株筛选
(1)溶液的配制
pH1.5人工胃液:参考中国药典的人工胃液配方:取稀盐酸16.4mL,加水约800mL及胃蛋白酶10g,搅匀后加水定容至1000mL,0.22um水系滤器过滤除菌即可。(稀盐酸为234mL浓盐酸加水稀释至1000mL)其余pH的人工胃液直接在pH1.5的人工胃液下调节pH即可。
3g/L的牛胆盐:在液体培养基中加入牛胆盐,使之浓度达到3g/L,高压灭菌。
(2)抗逆性检测
选取菌种活化24小时,取菌液5000r/min离心20min后加入等体积人工胃液和3g/L胆盐,置于37℃处理2h,对未处理的菌液及处理2h后的菌液进行活菌计数计算存活率(如附表1所示)。
步骤B:高产生物素菌株筛选
(1)待测菌液制备
从超低温冰箱中将步骤A筛选的菌种取出,待常温解冻后按2%比例接种于MRS肉汤培养基,37±1℃培养19~24小时后获得一级种子液。从一级种子液中取菌液按2%比例重新接种培养后获得二级种子液。由于菌种在冰箱保存时间不一致可能导致活性差异,故使用二级种子液按2%比例接种至MRS肉汤培养24小时获得待测菌液以减小菌种活性造成的差异。
(2)无菌发酵液的制备
将待检测菌液置于离心机以10000 rpm离心10分钟后取上清液,在无菌操作台中用0.22μm水系滤头过滤离心上清液,滤液为无菌发酵液。
(3)生物素含量检测
具体检测步骤参照GB 5009.259-2016 《食品安全国家标准 食品中生物素的测定》进行修改制定。
表1 乳酸菌抗逆性及产生物素含量
实施例2:益生菌发酵药食同源组合物
步骤A:发酵种子液制备
将高产生物素菌种分别按2%分别接种于甘蔗糖蜜培养基中,置于37℃静置培养24小时,获得发酵种子液。
作为优选,所述益生菌菌种包括鼠李糖乳杆菌、植物乳杆菌、格式乳杆菌、罗伊氏乳杆菌、长双歧杆菌、发酵乳杆菌、乳酸乳球菌、嗜酸乳杆菌、保加利亚乳杆菌中的一种或多种。更优选地,所述益生菌菌种包括植物乳杆菌、罗伊氏乳杆菌、发酵乳杆菌。
作为优选,所述甘蔗糖蜜培养基使用纯化水稀释至糖度为(2~10)Brix,使用115℃高压蒸汽灭菌30分钟。更优选地,所述稀释至糖度为(4.9~5.5)Brix。
步骤B:发酵基质制备
将药食同源组合物转移至发酵罐中,加入纯化水后使用纯蒸汽进行115℃,30分钟杀菌,冷却至常温后获得发酵基质。
作为优选,所述药食同源组合物为干燥粉碎后过50~200目筛的粉剂,栀子:百合:黄芪:党参的混合比例为(1~6):(1~6):(1~6):(1~6)。更优选地,所述干燥方式为冷冻干燥,粉碎后过100目筛,栀子:百合:黄芪:党参的混合比例为3:6:1:1。
作为优选,所述发酵基质为1质量份药食同源组合物加入(10~30)质量份纯化水混合。更优选为1质量份药食同源组合物加入19质量份纯化水混合。
步骤C:将步骤A制备发酵种子液按比例接种至步骤B制备发酵基质中,恒温搅拌发酵,发酵产物为一种补气固发益生菌发酵药食同源组合物饮料。
作为优选,所述发酵种子液的接种比例为1‰~20‰。
作为优选,所述恒温搅拌发酵为控温37℃,转速30rpm,发酵时间24小时。
实验例1:益生菌发酵药食同源组合物急毒试验
本实验主要针对本发明物的急毒效果通过小鼠灌胃试验进行测定;通过对雌、雄小鼠给以10000mg/kg·bw,连续观察14天,记录体重变化和死亡情况判断急毒效果。
实验对象为健康成年SPF级ICR小鼠20只,雌雄各半,由南京医科大学医药实验动物中心提供,生产许可证号SCXK(苏)2016-0002,合格证编号:NYD-L-2020110905。实验动物在温度为20~26℃、相对湿度为40%~70%的本中心屏障系统中饲养,实验动物使用许可证号:SYXK(苏)2020-0006。实验鼠维持饲料(Co60辐照)由江苏省协同医药生物工程有限责任公司提供,生产许可证:苏饲证(2019)01008。
采用限量法,设定剂量为10000 mg/kg•bw,每组10只动物,雌雄各半。动物检疫3d,试验时雌性体重为19.2~21.0g,雄性体重为19.9~22.3g,试验前动物禁食4h,自由饮水。样品比重1.008,以原液按10 mL/kg•bw,1次灌胃给样,实际剂量为10080 mg/kg•bw。全部动物连续观察14天,记录中毒表现及死亡情况。
灌胃给予ICR小鼠10080 mg/kg•bw本发明物。雌、雄性动物14d试验观察期间,饮食和活动正常,生长良好,未见中毒表现及异常体征,无动物死亡(见表1)。所以,本发明物的雌、雄性小鼠急性经口毒性试验LD50均为大于10080 mg/kg•bw,属实际无毒(结果如表2所示)。
表2 小鼠急性经口毒性试验结果
实验例2:益生菌发酵药食同源组合物促进小鼠重新生发
最近有研究表明,脱发的发生与肠道微生物菌群失调存在关联,而生物素是一种高度依赖细菌代谢产生的营养素,生物素的缺乏会导致头发断裂和严重的皮肤疾病。本发明物旨在通过补充高产生物素的益生菌以缓解由于肠道菌群失调引起的脱发同时,补充生物素改善发质。研究主要通过以下方案验证:
选取健康成年SPF级雌雄小鼠各20只,按照性别随机分成两组,每组10只雌性小鼠,10只雄性小鼠。使用万古霉素分别处理4周后,观察到所有小鼠背部毛发脱落,从各组随机取血液和粪便样品进行生物素含量测定。在保证正常供水给食的基础上停用万古霉素,A组小鼠使用本发明物灌胃处理,灌胃量为20mL/d·kg,B组小鼠使用生理盐水替换进行灌胃处理,灌胃量为20mL/d·kg。持续给药8周后,按0~10分对小鼠毛发进行视觉评分,将小鼠背部毛发刮下来后称重,取血液和粪便样品测定生物素含量(结果如表3所示)。
附表3 益生菌发酵药食同源饮料对小鼠生理指标影响
恢复饮食后,小鼠逐渐恢复生物素水平,给予本发明物后,小鼠体内生物素含量显著提高,重新生长毛发。同时发现本发明物对于雄性小鼠的重新生发效果明显优于雌性,可能针对男性的诱导重新生发效果优于女性。
Claims (7)
1.将高产生物素菌种分别按2%分别接种于甘蔗糖蜜培养基中,置于37℃静置培养24小时,获得发酵种子液。
2.权利要求1所述高产生物素菌种包括鼠李糖乳杆菌、植物乳杆菌、格式乳杆菌、罗伊氏乳杆菌、长双歧杆菌、发酵乳杆菌、乳酸乳球菌、嗜酸乳杆菌、保加利亚乳杆菌中的一种或多种。更优选地,所述益生菌菌种包括植物乳杆菌、罗伊氏乳杆菌、发酵乳杆菌。
3.权利要求1所述甘蔗糖蜜培养基使用纯化水稀释至糖度为(2~10)Brix,使用115℃高压蒸汽灭菌30分钟。更优选地,所述稀释至糖度为(4.9~5.5)Brix。
4.将药食同源组合物转移至发酵罐中,加入纯化水后使用纯蒸汽进行115℃,30分钟杀菌,冷却至常温后获得发酵基质。
5.权利要求4所述药食同源组合物为干燥粉碎后过50~200目筛的粉剂,栀子:百合:黄芪:党参的混合比例为(1~6):(1~6):(1~6):(1~6)。更优选地,所述干燥方式为冷冻干燥,粉碎后过100目筛,栀子:百合:黄芪:党参的混合比例为3:6:1:1。
6.权利要求4所述发酵基质为1质量份药食同源组合物加入(10~30)质量份纯化水混合。更优选为1质量份药食同源组合物加入19质量份纯化水混合。
7.将权利要求1制备发酵种子液按比例接种至权利要求4制备发酵基质中,恒温搅拌发酵。所述发酵种子液的接种比例为1‰~20‰,所述恒温搅拌发酵为控温37℃,转速30 rpm,发酵时间24小时。
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