CN112244291A - 一种蛹虫草口服液及其制备方法和应用 - Google Patents
一种蛹虫草口服液及其制备方法和应用 Download PDFInfo
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- CN112244291A CN112244291A CN202011107910.1A CN202011107910A CN112244291A CN 112244291 A CN112244291 A CN 112244291A CN 202011107910 A CN202011107910 A CN 202011107910A CN 112244291 A CN112244291 A CN 112244291A
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Abstract
本发明公开的是一种蛹虫草口服液及其制备方法和应用,口服液含蛹虫草菌丝体提取液,蛹虫草菌丝体提取液通过以下方法获得:配制培养基、菌丝体菌液培养、蛹虫草菌丝体菌液提取液,蛹虫草口服液在制备增强机体免疫力,助睡眠的保健食品或药品中的应用,本发明组方简单,活性组分含量高,口服液中虫草素、腺苷含量高,保留了蛹虫草滤液中的活性成分,零污染,能耗低,节省成本;不添加任何防腐剂,蛹虫草中含有虫草素成分,同时还能起到很好的防腐作用,原料来源多。
Description
技术领域
本发明属于保健品领域,具体涉及一种高虫草素、腺苷含量的蛹虫草口服液及其制备方法和应用。
技术背景
蛹虫草,属真菌门(Fungi)、子囊菌纲(Ascomycota)、肉座菌目(Hypocreales)、麦角菌科(Clavicipitaceae)、虫草属(Cordyceps)。因含有虫草素、虫草酸、腺苷及多糖等活性成分而具有抗肿癌,抗炎,抗病毒,增强机体免疫力等功效,是一种具有滋补作用的药食两用真菌,有很高的药用价值和营养价值,被越来越多的消费者所接受,需求量也逐年攀升。
虫草素(cordycepin)又名3'一脱氧腺苷,是Cuningham于1951年分离得到的一种腺苷类活性物质,分子式为C10H13N503。虫草素是蛹虫草中主要的有效成份之一,具有抗菌抗病毒、抗肿瘤、以及抗疲劳、抗衰老、抗炎提高人体免疫能力,抑制单胺氧化酶活性,具有较高的药用价值和广泛的药理作用。
腺苷(Adenosine)是核苷的一种,由核糖(呋喃核糖)与腺嘌呤的一部分组成,中间由β-N9-配糖键(β-N9-glycosidic bond)连结。腺苷在生物化学上扮演重要角色,包括以腺苷三磷酸(ATP)或腺苷双磷酸(ADP)形式转移能量,或是以环状腺苷单磷酸(cAMP)进行信号传递等。此外腺苷也是一种抑制性神经传导物(inhibitory neurotransmitter),可能会促进睡眠。在冬虫夏草中腺苷含量高于虫草素,腺苷也是冬虫夏草的质量标准(《中国药典》)。
虫草菌丝体中含有多种活性成分,如虫草素、核苷、虫草多糖、胆甾醇、真菌甾醇、麦角甾醇、腺嘌呤、尿嘧啶、蛋白质、多种氨基酸、硬脂酸、软脂酸、D-甘露醇、维生素、有机酸和微量元素,既可有效保护虫草这一珍贵资源,又不受气候、地理环境和虫草寄生条件严格的限制,适合于工业化大规模生产。然而利用虫草菌丝体制备食品、保健品、药物的产品多只注重虫草素、虫草酸等活性成分的提要,而忽略了作为冬虫夏草质量标准的腺苷含量。
发明内容
为解决上述技术问题,本发明的目的在于提供一种蛹虫草口服液及其制备方法和应用,该口服液虫草素、腺苷含量高,组分简单,不添加防腐剂,可以满足人们对蛹虫草相关产品日益增加的需求。
为实现上述目的,本发明采用了如下技术方案:
一种蛹虫草口服液,其特征在于该口服液含蛹虫草菌丝体提取液。
所述蛹虫草菌丝体提取液通过以下方法获得:
A:配制培养基
1)取适量桑叶粉、玉米渣中加入水,文火煮10-60min,冷却、沉淀后用纱布过滤,取滤液,即桑叶玉米提取液;
2)取桑叶玉米提取液,加入葡萄糖、蛋白胨、磷酸二氢钾、磷酸二氢钠和硫酸镁,搅拌溶解,加入水定容至所需体积,分装,121摄氏度、1.1Mpa下高压灭菌20-30mi冷却待用;
B:菌丝体菌液培养
接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h;
C:蛹虫草菌丝体菌液提取液
收集菌液,煮沸10min,冷却,沉淀后取上清,即获得蛹虫草菌丝体提取液。
将所述的蛹虫草菌丝体提取液过滤,巴氏灭菌,分装获得。
作为优选所述桑叶粉和玉米渣是将桑叶和玉米烘干至含水量10%以下后粉碎获得。
作为优选配制培养基各组分的浓度为:葡萄糖5-20g/L、桑叶玉米提取液20-200(桑叶玉米提取液对应的桑叶粉和玉米渣的总重量)g/L、蛋白胨5-20g/L、磷酸二氢钾1.2-1.5g/L、磷酸二氢钠0.8-1.2g/L、硫酸镁1g/L。
作为优选配制培养基各组分的浓度为:葡萄糖5-15g/L、桑叶玉米提取液30-150(桑叶玉米提取液对应的桑叶粉和玉米渣的总重量)g/L、蛋白胨5-20g/L、磷酸二氢钾1.5g/L、磷酸二氢钠1g/L、硫酸镁1g/L。
作为优选所述的水为蒸馏水或去离子水。
作为优选过滤孔径为0.1-0.45微米,巴氏灭菌10min。
一种蛹虫草口服液在制备增强机体免疫力,助睡眠的保健食品或药品中的应用。
有益效果:
1.本发明组方简单,活性组分含量高,口服液中虫草素最高可达0.25mg/10ml,腺苷含量最高可达0.29mg/10ml,
2.本发明很好地保留蛹虫草滤液中的活性成分,不经过复杂的提取过程,整个生产过程几乎没有废液排放,做到零污染,同时大大降低能耗,节省成本;
3.本发明不添加任何防腐剂,蛹虫草中含有虫草素成分,同时还能起到很好的防腐作用。
4.我国是农业和蚕业大国,培养菌丝体的培养基用到的玉米和桑叶产量高,容易获得,成本低廉。
5.本发明生产过程中产生的桑叶玉米残渣还以及菌丝体残渣均可以应用于食物、动物饲料以及食品添加剂。组分可以充分利用,无浪费。
附图说明
图1是本发明实施例5中活性物质检测HPLC图谱。
图中:1号峰为腺苷,2号峰为虫草素。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。本发明所用的试剂皆为食品级及以上。
实施例1
一种蛹虫草液体培养基及其制备方法,包括如下步骤:
1、培养基配制
分别将桑叶玉米烘干至含水分10%以下,粉碎备用;
称取10g/L桑叶粉、20g/L玉米渣,加入水中搅拌均匀,文火煮10-60min(优选30min),冷却后用8-16层纱布过滤(优选8层),收集滤渣,取滤液,将滤液补水至初始加入水的体积,混匀备用。
称(量)取培养基各组分,培养基的组成成分为:葡萄糖15g/L、磷酸二氢钾1.2g/L、磷酸二氢钠1.2g/L、桑叶玉米提取液30(桑叶玉米提取液对应的桑叶粉玉米渣的总重量)g/L、蛋白胨20g/L、硫酸镁1g/L。混匀各组分,121℃灭菌20min。
2、蛹虫草菌丝体的培养
在超净台中或无菌状态下接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,密度控制在3-5个菌球/mL,按体积比1:100接种,每种培养基做三个重复。在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h(优选96h)。
3、蛹虫草口服液的制备
收集菌液,煮沸10min,冷却,菌液3000rpm离心3-20min(优选5min),取上清,即获得蛹虫草菌丝体提取液。
将蛹虫草菌丝体提取液经孔径为0.1-0.45(优选0.22)微米滤膜过滤,滤液巴氏灭菌10min。分装获得蛹虫草口服液。
4、活性组分的测定
采用HPLC的方法测定获得的蛹虫草口服液的虫草素和腺苷含量,结果如表1所示。
实施例2
一种蛹虫草液体培养基及其制备方法,包括如下步骤:
1、培养基配制
分别将桑叶玉米烘干至含水分10%以下,粉碎备用;
称取10g/L桑叶粉、50g/L玉米渣,加入水中搅拌均匀,文火煮10-60min(优选30min),冷却后用8-16层纱布过滤(优选8层),收集滤渣,取滤液,将滤液补水至初始加入水的体积,混匀备用。
称(量)取培养基各组分,培养基的组成成分为:葡萄糖10g/L、磷酸二氢钾1.5g/L、磷酸二氢钠1g/L、桑叶玉米提取液60(桑叶玉米提取液对应的桑叶粉玉米渣的总重量)g/L、蛋白胨15g/L、硫酸镁1g/L。混匀各组分,121℃灭菌20min。
2、蛹虫草菌丝体的培养
在超净台中或无菌状态下接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,密度控制在3-5个菌球/mL,按体积比1:100接种,每种培养基做三个重复。在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h(优选96h)。
3、蛹虫草口服液的制备
收集菌液,煮沸10min,冷却,菌液3000rpm离心3-20min(优选5min),取上清,即获得蛹虫草菌丝体提取液。
将蛹虫草菌丝体提取液经孔径为0.1-0.45(优选0.22)微米滤膜过滤,滤液巴氏灭菌10min。分装获得蛹虫草口服液。
4、活性组分的测定
采用HPLC的方法测定获得的蛹虫草口服液的虫草素和腺苷含量,结果如表1所示。
实施例3
一种蛹虫草液体培养基及其制备方法,包括如下步骤:
1、培养基配制
分别将桑叶玉米烘干至含水分10%以下,粉碎备用;
称取15g/L桑叶粉、30g/L玉米渣,加入水中搅拌均匀,文火煮10-60min(优选30min),冷却后用8-16层纱布过滤(优选8层),收集滤渣,取滤液,将滤液补水至初始加入水的体积,混匀备用。
称(量)取培养基各组分,培养基的组成成分为:葡萄糖15g/L、磷酸二氢钾1.5g/L、磷酸二氢钠1g/L、桑叶玉米提取液45(桑叶玉米提取液对应的桑叶粉玉米渣的总重量)g/L、蛋白胨10g/L、硫酸镁1g/L。混匀各组分,121℃灭菌20min。
2、蛹虫草菌丝体的培养
在超净台中或无菌状态下接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,密度控制在3-5个菌球/mL,按体积比1:100接种,每种培养基做三个重复。在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h(优选96h)。
3、蛹虫草口服液的制备
收集菌液,煮沸10min,冷却,菌液3000rpm离心3-20min(优选5min),取上清,即获得蛹虫草菌丝体提取液。
将蛹虫草菌丝体提取液经孔径为0.1-0.45(优选0.22)微米滤膜过滤,滤液巴氏灭菌10min。分装获得蛹虫草口服液。
4、活性组分的测定
采用HPLC的方法测定获得的蛹虫草口服液的虫草素和腺苷含量,结果如表1所示
实施例4
一种蛹虫草液体培养基及其制备方法,包括如下步骤:
1、培养基配制
分别将桑叶玉米烘干至含水分10%以下,粉碎备用;
称取25g/L桑叶粉、50g/L玉米渣,加入水中搅拌均匀,文火煮10-60min(优选30min),冷却后用8-16层纱布过滤(优选8层),收集滤渣,取滤液,将滤液补水至初始加入水的体积,混匀备用。
称(量)取培养基各组分,培养基的组成成分为:葡萄糖15g/L、磷酸二氢钾1.5g/L、磷酸二氢钠1g/L、桑叶玉米提取液75(桑叶玉米提取液对应的桑叶粉玉米渣的总重量)g/L、蛋白胨15g/L、硫酸镁1g/L。混匀各组分,121℃灭菌20min。
2、蛹虫草菌丝体的培养
在超净台中或无菌状态下接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,密度控制在3-5个菌球/mL,按体积比1:100接种,每种培养基做三个重复。在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h(优选96h)。
3、蛹虫草口服液的制备
收集菌液,煮沸10min,冷却,菌液3000rpm离心3-20min(优选5min),取上清,即获得蛹虫草菌丝体提取液。
将蛹虫草菌丝体提取液经孔径为0.1-0.45(优选0.22)微米滤膜过滤,滤液巴氏灭菌10min。分装获得蛹虫草口服液。
4、活性组分的测定
采用HPLC的方法测定获得的蛹虫草口服液的虫草素和腺苷含量,结果如表1所示
实施例5
一种蛹虫草液体培养基及其制备方法,包括如下步骤:
1、培养基配制
分别将桑叶玉米烘干至含水分10%以下,粉碎备用;
称取50g/L桑叶粉、100g/L玉米渣,加入水中搅拌均匀,文火煮10-60min(优选30min),冷却后用8-16层纱布过滤(优选8层),收集滤渣,取滤液,将滤液补水至初始加入水的体积,混匀备用。
称(量)取培养基各组分,培养基的组成成分为:葡萄糖5g/L、磷酸二氢钾1.5g/L、磷酸二氢钠1g/L、桑叶玉米提取液150(桑叶玉米提取液对应的桑叶粉玉米渣的总重量)g/L、蛋白胨10g/L、硫酸镁1g/L。混匀各组分,121℃灭菌20min。
2、蛹虫草菌丝体的培养
在超净台中或无菌状态下接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,密度控制在3-5个菌球/mL,按体积比1:100接种,每种培养基做三个重复。在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h(优选96h)。
3、蛹虫草口服液的制备
收集菌液,煮沸10min,冷却,菌液3000rpm离心3-20min(优选5min),取上清,即获得蛹虫草菌丝体提取液。
将蛹虫草菌丝体提取液经孔径为0.1-0.45(优选0.22)微米滤膜过滤,滤液巴氏灭菌10min。分装,每瓶10ml,获得蛹虫草口服液。
4、活性组分的测定
采用HPLC的方法测定获得的蛹虫草口服液的虫草素和腺苷含量,结果如表1所示。
表1各实施例获得的蛹虫草口服液中腺苷和虫草素的含量(mg/10ml)
实施例 | 腺苷 | 虫草素 |
1 | 0.201 | 0.195 |
2 | 0.222 | 0.217 |
3 | 0.208 | 0.201 |
4 | 0.258 | 0.251 |
5 | 0.293 | 0.245 |
通过从实验结果可以总结出,本发明提供蛹虫草口服液,在未经任何浓缩和添加的前提下,富含虫草素和腺苷两种活性物质。本发明提供的口服液还可添加其它调味剂来增加口感。经试验表明,本发明提供的口服液具有提高免疫力、助睡眠的功效,可开发为保健食品和药物。
本发明提供的在制备桑叶玉米提取物过程中所得的残渣以及蛹虫草菌丝体菌液离心后的沉淀还可用于食品、保健食品、动物饲料和添加剂的制备。
以上所述实施例仅是为充分说明本发明而所举的较佳的实施例,本发明的保护范围不限于此。本技术领域的技术人员在本发明基础上所作的等同替代或变换,均在本发明的保护范围之内。
Claims (9)
1.一种蛹虫草口服液,其特征在于该口服液含蛹虫草菌丝体提取液。
2.如权利要求1所述的一种蛹虫草口服液,其特征在于:所述蛹虫草菌丝体提取液通过以下方法获得:
A:配制培养基
1)取适量桑叶粉、玉米渣中加入水,文火煮10-60min,冷却、沉淀后用纱布过滤,取滤液,即桑叶玉米提取液;
2)取桑叶玉米提取液,加入葡萄糖、蛋白胨、磷酸二氢钾、磷酸二氢钠和硫酸镁,搅拌溶解,加入水定容至所需体积,分装,121摄氏度、1.1Mpa下高压灭菌20-30mi冷却待用;
B:菌丝体菌液培养
接种活虫草菌体(菌种为Cordyceps militaris(L)Link),液体菌种,在温度22℃条件下避光静置培养24h,然后22℃、170rpm继续避光培养60-120h;
C:蛹虫草菌丝体菌液提取液
收集菌液,煮沸10min,冷却,沉淀后取上清,即获得蛹虫草菌丝体提取液。
3.如权利要求2所述的一种蛹虫草口服液,其特征在于:将获得的蛹虫草菌丝体提取液过滤,巴氏灭菌,分装获得。
4.如权利要求2所述的一种蛹虫草口服液,其特征在于:所述桑叶粉和玉米渣是将桑叶和玉米烘干至含水量10%以下后粉碎获得。
5.如权利要求2所述的一种蛹虫草口服液,其特征在于:配制培养基各组分的浓度为:葡萄糖5-20g/L、桑叶玉米提取液20-200g/L;蛋白胨5-20g/L、磷酸二氢钾1.2-1.5g/L、磷酸二氢钠0.8-1.2g/L、硫酸镁1g/L。
6.如权利要求2所述的一种蛹虫草口服液,其特征在于:配制培养基各组分的浓度为:葡萄糖5-15g/L、桑叶玉米提取液30-150g/L、蛋白胨5-20g/L、磷酸二氢钾1.5g/L、磷酸二氢钠1g/L、硫酸镁1g/L。
7.如权利要求2所述的一种蛹虫草口服液,其特征在于:所述水为蒸馏水或去离子水。
8.如权利要求3所述的一种蛹虫草口服液,其特征在于:过滤孔径为0.1-0.45微米,巴氏灭菌10min。
9.权利要求1-8任一项所述的一种蛹虫草口服液在制备增强机体免疫力,助睡眠的保健食品或药品中的应用。
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