CN105963251B - 一种桑黄口服液的制备方法及专用培养基 - Google Patents
一种桑黄口服液的制备方法及专用培养基 Download PDFInfo
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Abstract
本发明公开了一种桑黄口服液的制备方法及专用培养基,从根本上解决了用生物发酵技术填补野生桑黄不足的现状,还提高了口服液中桑黄主要成分的含量。培养基以天然植物为主,天然绿色无化学残留,微量元素多,营养全面;葡萄糖与蔗糖、白糖、红糖之一的二糖共同添加,让成分全面持久;3、用植物油来替代传统的化学消泡剂,既有营养,又无化学残留;4、口服液在生产和加工灌装过程中,全程不采用高温、干燥、萃取、破碎等传统或现代的物理或化学方法,不仅有效地保留了口服液中所有的活性成分,而且使活性成分不被损伤或被破坏;从制备摇瓶种至产品灌装都不添加任何防腐剂、增稠剂、稳定剂、着色剂等添加剂,该口服液有效成分含量高。
Description
技术领域
本发明涉及一种口服液的制备方法,具体是一种桑黄口服液的制备方法及专用培养基。
背景技术
桑黄是一种名贵珍稀的药用真菌,因寄生于桑树上,初长成时呈褐黄色而得名桑黄,桑黄分布在东亚地区,包括我国、日本、韩国、朝鲜、俄罗斯等。
桑黄的药用在我国从汉朝就已经开始,《本草纲目》介绍桑黄“利五脏,宣肠胃气,排毒气”。在日本、韩国,桑黄的知名度如同我国的桑黄。
桑黄的成分特别复杂,含有多糖、黑色素、过氧化酶、麦角甾醇、芳樟醇、三萜酸、脂肪酸类、芳香酸、原儿茶酸、丁香酸、咖啡酸、柚皮素、香豆素等成分。
桑黄有提高免疫力、诱导癌细胞自行凋亡、抑制癌细胞的增殖及转移、减轻化疗和放疗副作用的功能,有护肝、调整和降低血糖浓度、降血脂、防止动脉硬化、防止心脑血管病的发生的作用,有预防和治疗类风湿性关节炎、抑制尿酸、对痛风有良好效果的功效,对过敏及久治不愈的湿疹也有疗效。有关研究表明,桑黄及其提取物对人体无毒无害,即使长期大剂量服用亦无任何毒副作用。
目前已有一些关于桑黄口服液的专利,如福建农林大学在2008年申请的“一种桑黄保健口服液的制备方法”(CN100438882C)。以桑黄菌为出发菌株,进行液体发酵,获得桑黄发酵醪,然后将发酵醪中的菌丝用热水浸提法获得桑黄多糖的提取液,并与发酵液一起制成浓缩液,最后将浓缩液调配成适口性良好的保健口服液。后来有人研究了复方桑黄口服液及制备方法。但多数口服液的制备过程中采用化学合成培养基,而且在发酵过程中采用化学消泡剂处理发酵产生的泡沫,另外口服液中常会添加防腐剂、增稠剂、稳定剂、着色剂等添加剂。口服液生产灌装过程中,会采用高温、干燥、萃取、破碎等传统或现代的物理化学方法,会破坏活性成分。
发明内容
本发明的目的是提供一种桑黄口服液的制备方法及专用培养基,从根本上解决了用生物发酵技术填补野生桑黄不足的现状,还提高了口服液中桑黄主要成分的含量,为食品工业和制药行业,为人民的健康提供了新的保障和途径。
本发明的技术方案是这样实现的:一种桑黄口服液的制备方法,步骤如下:
1)制备摇瓶种:将培养基装入三角烧瓶内,装液量25-35%,封口后在121-123℃的温度下灭菌35分钟,待培养基温度降到20-28℃时,在无菌条件下将斜面菌种接入三角瓶内上摇床培养,当菌液微稠,菌液色泽淡黄色或橙褐色,菌丝球清晰可见,周边毛刺明显,镜检无杂菌时,备用;
2)发酵罐培养:将培养基装入发酵罐中,装液量为发酵罐总容量的55-75%,当罐温升至121-123℃、罐压0.11-0.13Mpa时,灭菌50min,然后通冷却水降温,待罐内温度降到30℃时停止,调整罐压0.05Mpa,进入培养状态;
将培养好的摇瓶种菌种接入罐内进行培养,接种量5-15%,罐温20-28℃,罐压0.04-0.05MPa,气体流量1.5m3/h,罐内培养36-48h时,取样检验;
继续培养5-7d,当菌球清晰可见,周边毛刺明显,菌液呈淡黄色、pH5-6左右、残糖量降至0.3%左右时,培养结束,进行分装。所述分装是在无菌无尘车间内,采用冷链专用灌装设备,将发酵液灌至灭过菌的瓶内。
进一步地,将步骤1)的摇瓶种接入种子罐放大培养,再扩接到发酵罐培养。
进一步地,所述的糖为葡萄糖与二糖的混合,其各占50%质量百分比。所述的二糖为蔗糖、白糖或者红糖。
进一步地,视产品需要或者工艺要求,分装前也可过滤除去菌球。
进一步地,步骤1)摇瓶培养基配方:以重量百分比计为:桑木0.5-1.5%、麸皮0.5-1.5%、玉米1.5-2.5%、黄豆1.5-2.5%、燕麦0.5-1.5%、糖1.5-2.5%,酵母粉1-2%,蜂花粉0.1-0.2%,磷酸二氢钾0.2%,余量为水。
进一步地,步骤1)和步骤2中培养基的pH值为5.5-7.5。
进一步地,所述步骤2)培养基的配方为:原料按重量百分比为:桑木0.5-1.5%,麸皮0.5-1.5%,玉米1.5-2.5%,黄豆1.5-2.5%,燕麦0.5-1.5%,糖1.5-2.5%,酵母粉1-2%,蜂花粉0.1-0.2%,磷酸二氢钾0.2%,植物油0.3-0.6%,余量为水。
进一步地,步骤1)中摇床转速180-240r/min,摇床培养是20-28℃下振荡培养7-10天。
进一步地,步骤1)和步骤2)中培养基的制备方法为:桑木锯末用筛滤去杂物煮汁过滤,树皮、树枝、树叶、麸皮直接煮汁过滤,玉米、黄豆、燕麦浸透无硬心后打浆过滤去渣,将过滤后的汁液一次加入其它辅料,按比例补足水,装入三角烧瓶内。
进一步地,步骤2)中,当罐温升至80℃时,先打开排气阀,排出冷空气,此时排气阀处于微开状态。
本发明的有益效果为:1、改变了以往发酵的化学合成培养基,而是以天然植物为主,天然绿色无化学残留,微量元素多,营养全面;2、传统配方中只添加葡萄糖这样的单糖,现改为葡萄糖与蔗糖、白糖、红糖之一的二糖共同添加,让成分全面持久;3、传统发酵中用消泡剂来处理发酵产生的泡沫,该技术用植物油来替代,既有营养,又无化学残留;4、口服液在生产和加工灌装过程中,全程不采用高温、干燥、萃取、破碎等传统或现代的物理或化学方法,不仅有效地保留了口服液中所有的活性成分,而且使活性成分不被损伤或被破坏;5、口服液中改变以往添加防腐剂、增稠剂、稳定剂、着色剂等添加剂的方法,从制备摇瓶种至产品灌装都不添加任何不符合国家要求的化学添加剂;6、该种方法生产的口服液经权威检测部门检测有效成分含量高;7、采用液体发酵的方法,缩短了生产周期,产量高,成分好。
具体实施方式
下面结合具体实施对本发明作进一步解释说明。
实施例1
1、摇瓶种配方:原料按重量的百分比为:桑木(锯末、树皮、树叶、树枝)0.5%,麸皮0.5%,玉米2.5%,黄豆2.5%,燕麦0.5%,糖2.5%(其中,糖为葡萄糖和蔗糖,各占50%),酵母粉1%,蜂花粉0.2%,磷酸二氢钾0.2%,余量为水,pH值为5.5。
2、方法:桑木锯末用筛滤去杂物煮汁过滤,树皮、树枝、树叶、麸皮直接煮汁过滤,玉米、黄豆、燕麦浸透无硬心后打浆过滤去渣,将过滤后的汁液一次加入其它辅料,按比例补足水,装入三角烧瓶内,装液量30%左右,将棉塞(把棉花用纱布包好做成的)或硅胶塞塞入瓶口,用牛皮纸包好后进行高压灭菌,排净冷空气后升温至121℃,计时35min,待表针归零时打开排气阀开启锅盖。当瓶中培养基温度降到25℃时,在无菌条件下用接种铲将斜面菌种接入三角瓶内,接完菌的三角瓶移入培养室上摇床培养,摇床转速200r/min,25℃下振荡培养8天,当菌液微稠,菌液色泽淡黄色或橙褐色,菌丝球清晰可见,周边毛刺明显,镜检无杂菌时,备用。
二、发酵罐培养
1、培养基:原料按重量的百分比为:桑木0.5%,麸皮1.5%,玉米2.5%,黄豆1.5%,燕麦0.5%打浆过滤,糖(其中糖为葡萄糖和蔗糖,各占50%)1.5%,酵母粉1%,蜂花粉0.1%,磷酸二氢钾0.2%,植物油0.6%,余量为水,pH值为5.5。制备方法同摇瓶培养基。
2、液体发酵培养方法和条件:将原料按比例称重,处理方法同摇瓶种,搅拌均匀,上料灭菌。装液量为发酵罐总容积的60%。当罐温升至80℃时,打开排气阀,排出冷空气,此时排气阀处于微开状态,罐温升至121-123℃、罐压0.12Mpa时,计时开始,灭菌50min,计时结束通冷却水降温,罐内温度降到30℃时停止降温,调整罐压0.05Mpa,进入培养状态。将培养好的摇瓶种或种子罐菌种接入罐内进行培养,接种量10%,罐温28℃,罐压0.04Mpa,气体流量1.5m3/h,罐内培养36h时,取样检验。继续培养6天,当菌球清晰可见,周边毛刺明显,菌液呈淡黄色、pH5-6左右、残糖量降至0.3%左右时,培养结束,经检验合格即可分装(分装时视产品需要或工艺要求保留菌球或滤去菌球)。
三、分装、旋盖、灯检、贴标、装箱
在国家规定的标准净化无菌无尘车间内,采用冷链专用灌装设备,将发酵液灌至30-150mL的灭过菌的瓶内,旋盖后进行灯检、贴标、装箱。
实施例2
1、摇瓶种配方:原料按重量的百分比为:桑木(锯末、树皮、树叶、树枝)1.0%,麸皮1.0%,玉米1.5%,黄豆2%,燕麦1.5%,糖1.5%(其中,糖为葡萄糖和红糖,各占50%),酵母粉2%,蜂花粉0.1%,磷酸二氢钾0.2%,余量为水,pH值为6.5。
2、方法:桑木锯末用筛滤去杂物煮汁过滤,树皮、树枝、树叶、麸皮直接煮汁过滤,玉米、黄豆、燕麦浸透无硬心后打浆过滤去渣,将过滤后的汁液一次加入其它辅料,按比例补足水,装入三角烧瓶内,装液量30%左右,将棉塞(把棉花用纱布包好做成的)或硅胶塞塞入瓶口,用牛皮纸包好后进行高压灭菌,排净冷空气后升温至123℃,计时35min,待表针归零时打开排气阀开启锅盖。当瓶中培养基温度降到28℃时,在无菌条件下用接种铲将斜面菌种接入三角瓶内,接完菌的三角瓶移入培养室上摇床培养,摇床转速200r/min,28℃下振荡培养7天,当菌液微稠,菌液色泽淡黄色或橙褐色,菌丝球清晰可见,周边毛刺明显,镜检无杂菌时,备用。
二、发酵罐培养
1、培养基:原料按重量的百分比为:桑木1.5%,麸皮1 %,玉米2%,黄豆2%,燕麦1%打浆过滤,糖(其中糖为葡萄糖和蔗糖,各占50%)2%,酵母粉1%,蜂花粉0.15%,磷酸二氢钾0.2%,植物油0.4%,余量为水,pH值为6.5。制备方法同摇瓶培养基。
2、液体发酵培养方法和条件:将原料按比例称重,处理方法同摇瓶种,搅拌均匀,上料灭菌。装液量为发酵罐总容积的70%。当罐温升至80℃时,打开排气阀,排出冷空气,此时排气阀处于微开状态,罐温升至123℃、罐压0.11Mpa时,计时开始,灭菌50min,计时结束通冷却水降温,罐内温度降到30℃时停止降温,调整罐压0.05Mpa,进入培养状态。将培养好的摇瓶种或种子罐菌种接入罐内进行培养,接种量10%,罐温28℃,罐压0.04Mpa,气体流量1.5m3/h,罐内培养36h时,取样检验。继续培养7天,当菌球清晰可见,周边毛刺明显,菌液呈淡黄色、pH5-6左右、残糖量降至0.3%左右时,培养结束,经检验合格即可分装(分装时视产品需要或工艺要求保留菌球或滤去菌球)。
三、分装、旋盖、灯检、贴标、装箱
在国家规定的标准净化无菌无尘车间内,采用冷链专用灌装设备,将发酵液灌至30-150mL的灭过菌的瓶内,旋盖后进行灯检、贴标、装箱。
实施例3
1、摇瓶种配方:原料按重量的百分比为:桑木(锯末、树皮、树叶、树枝)1.5%,麸皮1.5%,玉米2.5%,黄豆1.5%,燕麦1.5%,糖2.5%(其中,糖为葡萄糖和白糖,各占50%),酵母粉1%,蜂花粉0.2%,磷酸二氢钾0.2%,余量为水,pH值为7。
2、方法:桑木锯末用筛滤去杂物煮汁过滤,树皮、树枝、树叶、麸皮直接煮汁过滤,玉米、黄豆、燕麦浸透无硬心后打浆过滤去渣,将过滤后的汁液一次加入其它辅料,按比例补足水,装入三角烧瓶内,装液量30%左右,将棉塞(把棉花用纱布包好做成的)或硅胶塞塞入瓶口,用牛皮纸包好后进行高压灭菌,排净冷空气后升温至121℃,计时35min,待表针归零时打开排气阀开启锅盖。当瓶中培养基温度降到20℃时,在无菌条件下用接种铲将斜面菌种接入三角瓶内,接完菌的三角瓶移入培养室上摇床培养,摇床转速200r/min,20℃下振荡培养10天,当菌液微稠,菌液色泽淡黄色或橙褐色,菌丝球清晰可见,周边毛刺明显,镜检无杂菌时,备用。
二、发酵罐培养
1、培养基:原料按重量的百分比为:桑木0.5%,麸皮1.5%,玉米2.5%,黄豆1.5%,燕麦1.5%打浆过滤,糖(其中糖为葡萄糖和蔗糖,各占50%)2.5%,酵母粉1%,蜂花粉0.1%,磷酸二氢钾0.2%,植物油0.6%,余量为水,pH值为5.5。制备方法同摇瓶培养基。
2、液体发酵培养方法和条件:将原料按比例称重,处理方法同摇瓶种,搅拌均匀,上料灭菌。装液量为发酵罐总容积的55%。当罐温升至80℃时,打开排气阀,排出冷空气,此时排气阀处于微开状态,罐温升至121-123℃、罐压0.13Mpa时,计时开始,灭菌50min,计时结束通冷却水降温,罐内温度降到30℃时停止降温,调整罐压0.05Mpa,进入培养状态。将培养好的摇瓶种或种子罐菌种接入罐内进行培养,接种量10%,罐温28℃,罐压0.05Mpa,气体流量1.5m3/h,罐内培养36h时,取样检验。继续培养5天,当菌球清晰可见,周边毛刺明显,菌液呈淡黄色、pH5-6左右、残糖量降至0.3%左右时,培养结束,经检验合格即可分装(分装时视产品需要或工艺要求保留菌球或滤去菌球)。
三、分装、旋盖、灯检、贴标、装箱
在国家规定的标准净化无菌无尘车间内,采用冷链专用灌装设备,将发酵液灌至30-150mL的灭过菌的瓶内,旋盖后进行灯检、贴标、装箱。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明披露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (8)
1.一种桑黄口服液的制备方法,其特征在于,步骤如下:
1)制备摇瓶种:
将培养基装入三角烧瓶内,装液量25-35%,封口后在121-123℃的温度下灭菌35分钟,待培养基温度降到20-28℃时,在无菌条件下将斜面菌种接入三角瓶内上摇床培养,当菌液微稠,菌液色泽淡黄色或橙褐色,菌丝球清晰可见,周边毛刺明显,镜检无杂菌时,备用;其中,摇瓶培养基配方:以重量百分比计为:桑木0.5-1.5%、麸皮0.5-1.5%、玉米1.5-2.5%、黄豆1.5-2.5%、燕麦0.5-1.5%、糖1.5-2.5%,酵母粉1-2%,蜂花粉0.1-0.2%,磷酸二氢钾0.2%,余量为水;
2)发酵罐培养:
将培养基装入发酵罐中,装液量为发酵罐总容量的55-75%,当罐温升至121-123℃、罐压0.11-0.13Mpa时,灭菌50min,然后通冷却水降温,待罐内温度降到30℃时停止,调整罐压至0.05MPa,进入培养状态;其中培养基的配方为:原料按重量百分比为:桑木0.5-1.5%,麸皮0.5-1.5%,玉米1.5-2.5%,黄豆1.5-2.5%,燕麦0.5-1.5%,糖1.5-2.5%,酵母粉1-2%,蜂花粉0.1-0.2%,磷酸二氢钾0.2%,植物油0.3-0.6%,余量为水;
将培养好的摇瓶种菌种接入罐内进行培养,接种量5-15%,罐温20-28℃,罐压0.04-0.05MPa,气体流量1.5m3/h,罐内培养36-48h时,取样检验;
继续培养5-7天,当菌球清晰可见,周边毛刺明显,菌液呈淡黄色、pH5-6、残糖量降至0.3%时,培养结束,进行分装。
2.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,将步骤1)的摇瓶种接入种子罐放大培养,再扩接到发酵罐培养。
3.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,分装前过滤除去菌球。
4.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,所述的糖为葡萄糖与二糖的混合,其各占50%质量百分比,所述的二糖为蔗糖、白糖或者红糖。
5.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,培养基的pH值为5.5-7.5。
6.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,步骤1)中摇床转速180-240r/min,摇床培养是20-28℃下振荡培养7-10天。
7.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,步骤1)和步骤2)中培养基的制备方法为:桑木锯末用筛滤去杂物煮汁过滤,树皮、树枝、树叶、麸皮直接煮汁过滤,玉米、黄豆、燕麦浸透无硬心后打浆过滤去渣,将过滤后的汁液一次加入其它辅料,按比例补足水,装入三角烧瓶内。
8.根据权利要求1所述的桑黄口服液的制备方法,其特征在于,步骤2)中,当罐温升至80℃时,先打开排气阀,排出冷空气,此时排气阀处于微开状态。
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