CN114514973B - 一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法 - Google Patents
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法 Download PDFInfo
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Abstract
本发明公开了一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，属于发酵饮料领域，步骤包括称量、煎煮、混菌发酵、离心、调配灌装、灭菌。本发明产品解酒护肝功效强，复合饮料可显著提高有效成分如葛根素、大豆苷元、栀子苷、黄酮、多酚、二氢杨梅素的含量，从而促进人体对酒精的分解吸收，增强发酵葛根复合饮料的醒酒能力，还能起到滋养脾胃肝肾的功效。
Description
技术领域
本发明涉及发酵饮料技术领域，具体为一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法。
背景技术
酒和酒文化在我国占据着重要地位，饮酒已经深入我们生活中，但饮酒量与不良健康结局的发生风险呈“J型曲线”。全球因饮酒而造成死亡的人数超过273万人，占全球死亡总数的5.2％，其中因饮酒而导致肝硬化和癌症死亡的人数分别占有47.7％和4.2％。酒精性肝病(ALD)是导致人体患慢性肝病的主要原因，ALD包括一系列肝病，主要是从轻度脂肪变性开始，发展为脂肪性肝炎、纤维化和肝硬化。然而，随着时代的发展与进步，人们越来越重视自身健康问题，如何从日常饮食中防止疾病的发生成为人们迫切关注的话题。目前，有众多的具有解酒护肝功效的药品，但长期服用会具有一定的毒性。同样，与解酒功效相关保健品也愈发增多，但由于保健品的价格昂贵，消费者只能“量力而行”。因此，开发一种具有解酒护肝功效且价格亲民的食品为现实所需。
葛根为豆科葛属，《中国药典2010》记载葛根有通经活络，解酒毒之功能。枳椇子作为中医传统解酒毒的中草药，含有大量的二氢杨梅素、有机酸、葡萄糖、有机酸等，具有解酒毒、促进尿液排毒的功效。何会等发现枳椇子葛根中剂量复方能够降低醉酒小鼠血清中的乙醇、乙醛含量，降低醉酒率，能明显加快志愿者酒后的乙醇代谢速度。研究表明，葛花含有的皂角苷、异黄酮类等活性成分能有效地降低酒精在胃肠的吸收，而且可加速乙醛在人和大鼠体内的排除。栀子来源于茜草科植物栀子的干燥成熟果实，有泻火除烦、清热利湿、凉血解毒的功效。
虽已有葛根、葛花、枳椇子单独使用，或与其他药材联合使用进行解酒护肝作用，但在解酒方面尚未见到葛根、葛花、枳椇子、枇杷叶、栀子、铁皮石斛叶六味药材配伍。并且较多解酒饮料的研发均为多种中药材简单配伍，这会使药效不能很好地发挥作用。益生菌和中草药在许多方面有相得益彰的协同作用，利用益生菌发酵中药不仅可以节约中药资源，而且还可以促进其有效成分溶出，降低中药的毒副作用。而现在益生菌发酵主要采用的是单菌种发酵，复合菌种发酵的研究相对较少。本研究旨在通过多种传统中药材的混合制成水煎剂，经混菌发酵，得到一款具有解酒护肝功效的饮品，并探究其解酒的机制，为今后消费者提供酒前或者酒后饮品，从而降低由酒精导致的肝脏疾病机率。
发明内容
为解决上述问题，本发明的目的是提供一种具有解酒护肝功效的葛根复合发酵饮料。
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，包括以下步骤：
S1、称量：葛根30-50份、葛花10-20份、枳椇子5-15份、枇杷叶20-40份、栀子10-20份、铁皮石斛叶5-10份；
S2、煎煮：先将药食同源原料于冷水中浸泡30-60min，料水比1:50-1:200，浸泡完成后煎煮30-180min；
S3、混菌发酵：将煎煮好的原液和药材渣一起进行混菌发酵，混菌中乳酸芽孢杆菌：植物乳杆菌＝8:1-1:1，将复合菌种于生理盐水中活化后按1％-5％接种，白砂糖添加量为2％-6％，发酵温度为28℃-36℃，发酵天数为5-14天；
S4、离心:发酵完毕后，将发酵液4000rpm/min离心15min，取上清备用；
S5、调配灌装:向上清液加入山梨酸钾、异维生素C钠、乙基麦芽酚、蜂蜜等进行调配，无氧灌装，得灌装饮料；
S6、灭菌:将灌装饮料放入水浴式灭菌锅内灭菌，于121℃恒温条件下灭菌15-40min，冷却至室温后，得终产品。
优选的，步骤S1中，葛根30份、葛花10份、枳椇子8份、枇杷叶20份、栀子10份、铁皮石斛叶5份。
优选的，步骤S2中，浸泡时间为30min，煎煮时间为60min，料水比1:100。
优选的，步骤S3中，乳酸芽孢杆菌：植物乳杆菌＝4:1。
优选的，步骤S3中，复合菌种于生理盐水中活化后按2％接种，白砂糖添加量为3％，发酵温度为29℃，发酵天数为8天。
本发明的有益效果：
①由对照案例的实验结果表明，添加了铁皮石斛叶和枇杷叶的解酒饮料的醒酒护肝功效比不含这两种中药材的解酒饮料要好，说明添加了这两味药材可起到协同增效的作用，使解酒护肝功效增强。
②益生菌发酵葛根复合饮料可显著提高有效成分如葛根素、大豆苷元、栀子苷、黄酮、多酚、二氢杨梅素的含量，从而促进人体对酒精的分解吸收，增强发酵葛根复合饮料的醒酒能力。相关研究表明葛根素、大豆苷元可降低乙醇的吸收、提高乙醛脱氢酶(ALDH)和乙醇脱氢酶(ADH)的活性，促进乙醇在体内的代谢，从而达到醒酒的目的。而黄酮和多酚具有较强的清除自由基的能力，可减轻肝脏损伤。
③乳酸菌复合发酵过程中可产生乳酸等风味成分赋予葛根发酵复合饮料特殊的发酵食品香气，可改善中药材煎煮后的苦涩味，增加葛根复合饮料的口感，同时还能起到滋养脾胃肝肾的功效。并且药食同源中药材经过微生物的发酵作用，不仅能使有效物质得以溶出，还会产生许多有益于身体平衡的功能因子，将发酵手段应用于饮料制备，开发具有一定功能的以药食两用中药为原料的新型发酵饮料，对于饮料市场结构和产品升级具有重要意义。
④葛根复合饮料采用的是复合菌种发酵。不同的微生物具有不同的酶系，采用多菌种混合发酵，可以使***的酶系多样化和互补，对于成分复杂的中药生物转化效率更高，多菌种混合发酵还可使植物组织内的功效成分及营养成分得到充分的分解、溶出和利用，提高中药材的综合利用率和生物转化率，从而提高葛根发酵复合饮料的醒酒能力。
⑤调配时，还加入了蜂蜜，蜂蜜中含有丰富的果糖。蜂蜜进入人体时，能快速的被吸收，增加人体的解毒功能，能加速身体里酒精的消解，排出体外，达到解酒的效果。
附图说明
图1：各对比例和实施例对醉酒大鼠血醇浓度影响；
图2：各对比例和实施例中大鼠血清中AST和ALT酶活力；
图3：各对比例和实施例中大鼠血清中CAT和AKP活力；
图4：各对比例和实施例中大鼠血清中Alb含量；
图5：各对比例和实施例的大鼠血清中ADH和ALDH活性；
图6：各对比例和实施例的大鼠肝切片效果图。
其中：
图2中：与对照组比较**P＜0.01，***P＜0.001；与模型组比较##P＜0.01，###P＜0.001；
图3中：与对照组比较**P＜0.01，***P＜0.001；与模型组比较##P＜0.01，###P＜0.001；
图4中：与对照组比较**P＜0.01，***P＜0.001；与模型组比较##P＜0.01，###P＜0.001；
图5中：与对照组比较**P＜0.01，***P＜0.001；与模型组比较##P＜0.01，###P＜0.001。
具体实施方式
下面将结合本发明实施例中的附图表，对本发明的技术方案进行进一步详细的描述，但本发明并不局限于下述实施例。基于本发明中的实施例，在本领域中的普通技术人员没有做出创造性劳动的前提下所取得的所有其他实施例，都属于本发明的保护范围。
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，包括以下步骤：
S1、称量：葛根30份、葛花10份、枳椇子8份、枇杷叶20份、栀子10份、铁皮石斛叶5份；
S2、煎煮：先将药食同源原料于冷水中浸泡30min(料液比1：100)，浸泡完成后煎煮60min；
S3、混菌发酵：取煎煮好的原液和药材渣一起进行混菌发酵(乳酸芽孢杆菌：植物乳杆菌＝4:1)，将复合菌种于生理盐水中活化后按2％接种，白砂糖添加量为3％，发酵温度为29℃，发酵天数为8天；
S4、离心:发酵完毕后，将发酵液4000rpm/min离心15min，取上清备用；
S5、调配灌装:加入山梨酸钾、异维生素C钠、乙基麦芽酚、蜂蜜等进行调配，无氧灌装，得灌装饮料；
S6、灭菌:将灌装饮料放入水浴式灭菌锅内灭菌，于121℃恒温条件下灭菌15min，冷却至室温后，得终产品。
实施例2：
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，包括以下步骤：
S1、称量：葛根30份、葛花10份、枳椇子8份、枇杷叶20份、栀子10份、铁皮石斛叶5份；
S2、煎煮：先将药食同源原料于冷水中浸泡30min(料液比1：200)，浸泡完成后煎煮90min；
S3、混菌发酵：取煎煮好的原液和药材渣一起进行混菌发酵(乳酸芽孢杆菌：植物乳杆菌＝2:1)，将复合菌种于生理盐水中活化后按2％接种，白砂糖添加量为3％，发酵温度为29℃，发酵天数为8天；
S4、离心:发酵完毕后，将发酵液4000rpm/min离心15min，取上清备用；
S5、调配灌装:加入山梨酸钾、异维生素C钠、乙基麦芽酚、蜂蜜等进行调配，无氧灌装，得灌装饮料；
S6、灭菌:将灌装饮料放入水浴式灭菌锅内灭菌，于121℃恒温条件下灭菌15min，冷却至室温后，得终产品。
实施例3：
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，包括以下步骤：
S1、称量：葛根30份、葛花10份、枳椇子8份、枇杷叶20份、栀子10份、铁皮石斛叶5份；
S2、煎煮：先将药食同源原料于冷水中浸泡30min(料液比1：50)，浸泡完成后煎煮120min；
S3、混菌发酵：取煎煮好的原液和药材渣一起进行混菌发酵(乳酸芽孢杆菌：植物乳杆菌＝1:1)，将复合菌种于生理盐水中活化后按2％接种，白砂糖添加量为3％，发酵温度为29℃，发酵天数为8天；
S4、离心:发酵完毕后，将发酵液4000rpm/min离心15min，取上清备用；
S5、调配灌装:加入山梨酸钾、异维生素C钠、乙基麦芽酚、蜂蜜等进行调配，无氧灌装，得灌装饮料；
S6、灭菌:将灌装饮料放入水浴式灭菌锅内灭菌，于121℃恒温条件下灭菌15min，冷却至室温后，得终产品。
对比例1：
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，包括以下步骤：
步骤S1的原料无枇杷叶和铁皮石斛叶，且无步骤S3，其余与实施例1相同。
对比例2：
一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，包括以下步骤：
除无步骤S3以外，其余与实施例1相同。
对比例3：为市售的一款解酒产品。
实验例1：葛根复合发酵饮料的感官评定
表1各对比例和实施例感官评价的评判标准
表2各对比例和实施例感官评定结果
经对各对比例和实施例进行感官评定，实施例1的感官评分最高，甜度、酸度、香味、口感均为最佳。
实验例2：发酵前后葛根饮料化学成分的变化
表3各对比例和实施例的化学成分变化
实验例3：葛根复合发酵饮料的降血醇作用
1.1实验动物
SPF级SD雄性大鼠64只，180-200g，随机分为8组，每组8只，分别为模型组；阳性对照组；对比案例1、2、3；实施案例1、2、3。动物由广东省医学实验动物中心提供，许可证号：SCXK(粤)2018-0002。
1.2实验方法
建立急性酒精性肝损伤模型模型。适应性喂养一周后，各组大鼠灌胃相应的样品，2次/天，间隔4h，末次给药30min后，按照大鼠体重灌胃给予43度红星二锅头白酒6mL/kg，正常组给予生理盐水。酒精灌胃后的1h，2h，4h三个时间段收集大鼠的血液，分离血清测血醇的浓度。
1.3实验结果
1.3.1葛根复合发酵饮料的降血醇作用
由图1可知，与模型组相比，实施案例1、实施案例2、实施案例3能够明显抑制醉酒大鼠血液乙醇浓度的升高，并且实施案例1的效果最优，其能够更快将血醇浓度降低至较低水平。说明实施案例1可以起到很好的降血醇作用。
实验例4：各对比例和实施例大鼠的解酒机制研究
1.1实验动物
SPF级SD雄性大鼠90只，180-200g，按照体重随机分为9组，每组10只，分别为正常组；模型组；阳性对照组；对照案例1、2、3；实施案例1、2、3。大鼠由广东省医学实验动物中心提供，许可证号：SCXK(粤)2018-0002。
1.2实验方法
样品组的大鼠按照1mL/100g灌胃相应的样品，1天一次，持续7d，末次灌胃样品4h后，继续灌胃12mL/kg的43度红星二锅头白酒。禁食不禁水8h后处死大鼠。采集血液和肝脏。血液用生化管收集后冷冻离心，解剖取肝脏，用0.9％生理盐水冲洗，用滤纸吸干多余水分后称肝脏重量。肝脏用10％中性***固定，石蜡包埋，切成5μm的切片，苏木精伊红(H&E)染色观察肝组织病理学特征。
动物血液在4℃下离心，以4000r/min转速下离心10min。分离上清得到的淡黄色液体，即为血清，于-80℃条件下储存，待测。
1.3实验结果
1.3.1各对比例和实施例对大鼠体重和肝脏的影响
肝脏遭受酒精损伤时，会呈现细胞肿大情况，同时在脂肪代谢过程中，还易出现代谢紊乱现象，使脂肪堆积在肝细胞中，造成肝脏整体质量和体积的增加，肝脏指数异常升高。由表4可知，除正常组外，模型组的肝脏指数大于其他组别，说明模型组大鼠的肝脏受到了一定损伤。正常组高的原因可能与其正常生长导致体重差异造成的，对照例和实施案例组的肝脏指数相较于模型组均显著降低，且实施案例1降低水平最显著(P＜0.01)。表明实施案例1对酒精作用下的肝脏有一定的保护效果。
表4各对比例和实施例大鼠体重和肝重的变化
注：与对照组比较**p＜0.01；与模型组比较##p＜0.01。
1.3.2各对比例和实施例对大鼠血清AST、ALT活力影响
由酒精介导的氧化应激引发的肝功能异常，导致血清中天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)等肝酶的释放、脂质过氧化、抗氧化功能丧失。因此，AST、ALT酶活性的大小可以用于衡量肝功能的受损程度。如图2所示，酒精建模后，模型组血清中AST活力相较于对照组极显著升高(P＜0.001)。对照例和实施例大鼠血清中ALT、AST活力相较于模型组均有不同程度的降低(P＜0.05、P＜0.01或P＜0.001)，并且实施案例1降低最显著。由此可见，实施案例1能有效减轻因酒精作用导致的ALT、AST水平异常升高，对酒精性肝损伤有一定的保护效果。
1.3.3各对比例和实施例对大鼠血清氧化应激酶影响
越来越多的证据表明，氧化能力在酒精性肝损伤中扮演着重要的作用。氧化***最重要的是酶，如SOD，CAT，GSH-Px等，这些酶能够保护机体免受活性氧的破坏，并且急性酒精中毒能够导致大鼠肝脏中AKP活力升高和CAT酶活力降低，因此，评价体内酶活力大小能够反映机体抗氧化能力的强弱以及葛根发酵饮料是否增强其机体内的抗氧化能力。如图3所示，与正常组相比，模型组中AKP活力显著升高，CAT活力显著下降，且达到极显著水平，说明急性酒精中毒模型成功。而实施案例1能够降低由酒精导致的AKP活力升高，其降低效果达到极显著水平，并且能在一定程度上能够提高CAT的活力。说明实施案例1可以降低肝脏中AKP的氧化应激能力和提高CAT的活力来达到保肝的效果，提示着葛根发酵复合饮料可以缓解氧化应激损伤。
1.3.4各对比例和实施例对大鼠血清白蛋白的影响
一般正常人的白蛋白(Alb)在60-80g/L之间，假如超出这个范围，说明肝脏有一定的受损。因此，血清中白蛋白含量在一定程度上能够反映大鼠由急性酒精性中毒导致的肝炎情况。如图4所示，与正常组相比，模型组Alb含量显著上升(P＜0.001)，说明急性酒精中毒对血清中白蛋白有显著影响。与模型组相比，对照例和实施例大鼠的Alb含量均有不同程度的下降，并且实施案例1大鼠的Alb含量下降最显著(P＜0.001)，预示着该发酵复合饮料能够减少急性酒精中毒导致的肝炎的发生。
1.3.5各对比例和实施例对大鼠血清中ADH、ALDH活力的影响
对于饮酒大鼠，部分酒精被血管吸收并通过血液中的乙醇脱氢酶氧化，再被乙醛脱氢酶氧化而代谢，酒精的代谢非常依赖于乙醇脱氢酶和乙醛脱氢酶的活性，因此评价ADH和ALDH对酒精性肝损伤有积极意义。如图5所示，与正常组比较，模型组大鼠血清中ADH，ALDH活性降低，而给予灌胃各对照例和实施例样品可增加饮酒大鼠血清中ADH，ALDH活性。其中实施案例中的案例1显著增强ADH，ALDH的活性。由此可得该发明可通过增强大鼠血清中ADH和ALDH的活性来快速代谢酒精，使酒精含量降低，达到护肝的作用。
1.3.6各对比例和实施例对大鼠肝脏细胞的影响
如图6所示，空白对照组大鼠肝细胞结构正常，形态规整，细胞核呈圆形，整体细胞染色均匀，间隔清晰，未见变性坏死细胞和炎细胞浸润现象。模型组大鼠肝组织出现严重病变，组织结构紊乱，中央静脉显著扩大且严重变形，肝细胞肿胀甚至发生崩解，出现成片坏死现象，细胞核碎裂、溶解严重，胞浆稀松，细胞间间隙模糊，可见小泡性脂变，且炎症细胞浸润现象严重。各对照案例和实施案例的大鼠肝细胞组织损伤得到显著改善，细胞核破裂现象减少、肝细胞肿胀崩解程度减轻，炎性浸润和空泡变性等情况都有所减轻，表明葛根复合发酵饮料可有效改善急性酒精性肝损伤。
对于本领域技术人员而言，显然本发明不限于上述示范性实施例的细节，而且在不背离本发明的精神或基本特征的情况下，能够以其他的具体形式实现本发明。因此，无论从哪一点来看，均应将实施例看作是示范性的，而且是非限制性的，本发明的范围由所附权利要求而不是上述说明限定，因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。
此外，应当理解，虽然本说明书按照实施方式加以描述，但并非每个实施方式仅包含一个独立的技术方案，说明书的这种叙述方式仅仅是为清楚起见，本领域技术人员应当将说明书作为一个整体，各实施例中的技术方案也可以经适当组合，形成本领域技术人员可以理解的其他实施方式。
Claims (4)
1.一种具有醒酒护肝功效的葛根复合发酵饮料的制备方法，其特征在于，包括以下步骤：
S1、称量：葛根30份、葛花10份、枳椇子8份、枇杷叶20份、栀子10份、铁皮石斛叶5份；
S2、煎煮：先将药食同源原料于冷水中浸泡30-60min，料水比1:50-1:200，浸泡完成后煎煮30-180min；
S3、混菌发酵：将煎煮好的原液和药材渣一起进行混菌发酵，混菌中乳酸芽孢杆菌：植物乳杆菌＝8:1-1:1，将复合菌种于生理盐水中活化后按1％-5％接种，白砂糖添加量为2％-6％，发酵温度为28℃-36℃，发酵天数为5-14天；
S4、离心：发酵完毕后，将发酵液4000rpm离心15min，取上清备用；
S5、调配灌装：向上清液加入山梨酸钾、异维生素C钠、乙基麦芽酚、蜂蜜进行调配，无氧灌装，得灌装饮料；
S6、灭菌：将灌装饮料放入水浴式灭菌锅内灭菌，于121℃恒温条件下灭菌15-40min，冷却至室温后，得终产品。
2.根据权利要求1所述的具有醒酒护肝功效的葛根复合发酵饮料的制备方法，其特征在于，步骤S2中，浸泡时间为30min，煎煮时间为60min，料水比1:100。
3.根据权利要求1所述的具有醒酒护肝功效的葛根复合发酵饮料的制备方法，其特征在于，步骤S3中，乳酸芽孢杆菌：植物乳杆菌＝4:1。
4.根据权利要求1所述的具有醒酒护肝功效的葛根复合发酵饮料的制备方法，其特征在于，步骤S3中，复合菌种于生理盐水中活化后按2％接种，白砂糖添加量为3％，发酵温度为29℃，发酵天数为8天。
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