CN107712724A - 一种银杏粉脱毒的方法 - Google Patents
一种银杏粉脱毒的方法 Download PDFInfo
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- CN107712724A CN107712724A CN201711118551.8A CN201711118551A CN107712724A CN 107712724 A CN107712724 A CN 107712724A CN 201711118551 A CN201711118551 A CN 201711118551A CN 107712724 A CN107712724 A CN 107712724A
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Abstract
本发明公开了一种银杏粉脱毒的方法,属于脱毒处理领域。本发明利用高压常温、处理液对银杏粉进行预处理,同时从米糠中提取一种能够去除有毒物质的菌液,并与百合、山楂核、燕麦进行混合,由于百合、山楂核、燕麦都具备一定保健功效,一方面提高了银杏粉的营养价值,另一方面并通过上层滤网滴入,从而避免菌液直接接触银杏粉,同时也能为银杏粉提供活性物质,此外,再经微波提取,收集营养成分的同时进行去毒,以及通过活性炭吸附剂对毒素和杂质进行吸附的四重脱毒工序,从而确保银杏粉的安全。本发明解决了目前银杏粉大多需要进行脱毒来降低毒性成分,但大多方法脱毒方法时间较长,毒素去除不彻底,且容易造成活性物质损失较多的问题。
Description
技术领域
本发明属于脱毒处理领域,具体涉及一种银杏粉脱毒的方法。
背景技术
银杏粉,是由银杏研磨的粉状物质,有降低血清胆固醇、促进脑和四肢血液循环的能力。中老年人常遭受腿、脚、手供血不足所引起的痛苦。研究表明,银杏能保动脉、静脉和毛细血管免受伤害,并增强它们的强度和弹性。银杏俗称白果,是我国特有的珍稀果品。银杏果由肉质外种皮、硬质中种皮、膜质内种皮、种仁组成。银杏在我国有着悠久的食用药用历史,我国人民在长期对银杏的使用中积累了丰富宝贵的历史经验。据《本草纲目》记载:银杏具有“熟食温肺、益气、定喘嗽、缩小便、止白浊;生食降痰、消毒杀虫等药用功能。中医素以银杏种仁治疗支气管哮喘、慢性气管炎、肺结核、淋浊、小儿遗尿以及疮、癣等疾病。经常食用白果,可以滋阴养颜抗衰老,扩张微血管,促进血液循 环,使人肌肤、面部红润,精神焕发,延年益寿,是老幼皆宜的保健食品和款待国宾上客的特 制佳肴。种仁中的黄酮甙、苦内脂对脑血栓、老年性痴呆、高血压、高血脂、冠心病、动脉硬化、脑功能减退等疾病还具有特殊的预防和治疗效果。近年来,银杏的保健和药用价值得到广泛认可,存在较大的需求市场。
随着银杏作为地方特色产业在各地推广种植,我国银杏产量激增,银杏资源占世界的90%左右,市场开发潜力巨大,前景广阔。目前银杏粉也是多种多样,但因银杏中含氢氰酸、银杏酚酸、白果酚酸等毒性成分,严重影响该类产品的市场前景,去除毒素制备安全可食用的银杏产品成为了行业的关键点,银杏叶提取物标准规定银杏酸含量不超过10ppm。银杏中主要活性成分为银杏黄酮类和银杏内酯类物质。目前去除毒素的方法主要有:采用简单加热方法,温度高时间长容易破坏活性物质;采用甲醇、石油醚等有机溶剂多次萃取银杏叶,溶剂成本消耗较高;采用发酵法降低银杏叶和银杏果的毒性成分,发酵时间较长且毒素去除不彻底;采用强碱法脱酸,活性物质损失较多。因此,发明出一种高效去毒的方法迫在眉睫。
发明内容
本发明所要解决的技术问题:针对目前银杏粉大多需要进行脱毒来降低毒性成分,但大多方法脱毒方法时间较长,毒素去除不彻底,且容易造成活性物质损失较多的问题,提供一种银杏粉脱毒的方法。
为解决上述技术问题,本发明采用如下所述的技术方案是:
一种银杏粉脱毒的方法,其特征在于,该制备方法包括如下步骤:
(1)将银杏粉放入高压容器中处理,在常温下升压至8~10MPa,进行搅拌30~40min,1~2min内降压至常压,得处理后的银杏粉,将处理后的银杏粉、处理液进行混合,浸泡1~2h,得浸泡后的银杏粉,备用;
(2)将米糠、液体培养基在温度23~28℃下振荡培养5~6d,得培养液,将培养液用接种环划线到PDA培养基上,在温度28~30℃的温度下培养1~2d,用灭菌的接种环挑取PDA培养基的单菌落接种于种子培养基中摇床振荡扩大培养2~3d,挑取菌径最大的菌落,并接种于种子发酵培养基在温度28~30℃摇床中振荡培养30~36h,得培养液a,将培养液a用四层纱布过滤,得滤液,将滤液进行离心10~20min,得上清液;
(3)按重量份数计,取10~15份上清液、7~8份百合、3~5份山楂核、3~4份燕麦进行混合,并进行搅拌10~15min,得混合物,先将步骤(1)中备用的浸泡后的银杏粉放入发酵罐的底部,在浸泡后得银杏粉上面放一层滤网,再将混合物倒入发酵罐中,在温度28~32℃下发酵10~15min,去除滤网,过滤,得发酵物;
(4)将发酵物、水进行混合,浸泡1~2h,再进行微波提取,得提取液,向提取液中加入发酵物质量10~20%的活性炭,30~40min后过滤,得滤液a,将滤液a进行减压浓缩,浓缩至滤液体积80~90%,再进行冷冻干燥,得干燥物,将干燥物进行粉碎,即完成银杏粉脱毒方法。
所述步骤(1)中处理后的银杏粉、处理液的质量比1:5~6。
所述步骤(1)中处理液是按质量比1:1:40~50,将柠檬酸溶液、盐酸溶液、水进行混合,得混合物,即得处理液。
所述步骤(2)中米糠、液体培养基的质量比1:7~8。
所述步骤(2)中液体培养基是按重量份数计,取40~50份水、15~20份葡萄糖、5~8份蛋白胨、2~4份酵母浸粉、1~2份磷酸氢二钾、0.5~1份硫酸镁进行混合,高温灭菌,即得液体培养基。
所述步骤(2)中PDA培养基是按重量份数计,取70~80份水、20~30份马铃薯 、2~3份葡萄糖 、1~2份琼脂进行混合,高温灭菌,即得PDA培养基。
所述步骤(2)中种子培养基是按重量份数计,取50~60份水、15~20份琼脂、10~15份酵母膏、8~10份葡萄糖、5~7份蛋白胨、2~4份氯化钠进行混合,高温灭菌,即得种子培养基。
所述步骤(2)中种子发酵培养基是按重量份数计,取70~80份水、50~60份柠檬酸氢二铵、30~40份可溶性淀粉、30~40份蔗糖、8~10份磷酸氢二铵、0.5~1份氯化钙、0.5~1份硫酸铁、0.5~1份硫酸镁进行混合,高温灭菌,即得种子培养基。
所述步骤(3)中步骤(1)中备用的浸泡后的银杏粉、混合物的质量比1:8~10。
所述步骤(4)中发酵物、水的质量比1:3~4。
本发明与其他方法相比,有益技术效果是:本发明先利用高压常温、处理液对银杏粉进行预处理,由于处理液是包括柠檬酸溶液、盐酸溶液,利用微酸性环境以及高压状态下将银杏粉中的有毒物质的溶出和分解,从而为后续发酵提供有利条件,且柠檬酸可作为护色剂,从而保护银杏粉的色泽和营养成分,同时从米糠中提取一种能够去除有毒物质的菌液,并与百合、山楂核、燕麦进行混合,由于百合、山楂核、燕麦都具备一定保健功效,一方面提高了银杏粉的营养价值,另一方面并通过上层滤网滴入,从而避免菌液直接接触银杏粉,同时也能为银杏粉提供活性物质,此外,再经微波提取,收集营养成分的同时进行去毒,以及通过活性炭吸附剂对毒素和杂质进行吸附的四重脱毒工序,从而确保银杏粉的安全,本发明银杏粉最后通过冷冻干燥,从而有效保留了银杏粉有效成分的活性,可以使银杏粉长期保存备用。
具体实施方式
处理液是按质量比1:1:40~50,将柠檬酸溶液、盐酸溶液、水进行混合,得混合物,即得处理液。
液体培养基是按重量份数计,取40~50份水、15~20份葡萄糖、5~8份蛋白胨、2~4份酵母浸粉、1~2份磷酸氢二钾、0.5~1份硫酸镁进行混合,高温灭菌,即得液体培养基。
PDA培养基是按重量份数计,取70~80份水、20~30份马铃薯 、2~3份葡萄糖 、 1~2份琼脂进行混合,高温灭菌,即得PDA培养基。
种子培养基是按重量份数计,取50~60份水、15~20份琼脂、10~15份酵母膏、8~10份葡萄糖、5~7份蛋白胨、2~4份氯化钠进行混合,高温灭菌,即得种子培养基。
种子发酵培养基是按重量份数计,取70~80份水、50~60份柠檬酸氢二铵、30~40份可溶性淀粉、30~40份蔗糖、8~10份磷酸氢二铵、0.5~1份氯化钙、0.5~1份硫酸铁、0.5~1份硫酸镁进行混合,高温灭菌,即得种子培养基。
一种银杏粉脱毒的方法,该制备方法包括如下步骤:
(1)将银杏粉放入高压容器中处理,在常温下升压至8~10MPa,并以300~400r/min进行搅拌30~40min,1~2min内降压至常压,得处理后的银杏粉,按质量比1:3~4,将处理后的银杏粉、处理液进行混合,浸泡1~2h,得浸泡后的银杏粉,备用;
(2)按质量比1:3~4,将米糠、液体培养基在温度23~28℃下振荡培养5~6d,得培养液,将培养液用接种环划线到PDA培养基上,在温度28~30℃的温度下培养1~2d,用灭菌的接种环挑取PDA培养基的单菌落接种于种子培养基中,以100~120r/min摇床振荡扩大培养2~3d,挑取菌径最大的菌落,并接种于种子发酵培养基在温度28~30℃摇床中100~120r/min振荡培养30~36h,得培养液a,将培养液a用四层纱布过滤,得滤液,将滤液以1500~2000r/min下离心10~20min,得上清液;
(3)按重量份数计,取10~15份上清液、7~8份百合、3~5份山楂核、3~4份燕麦进行混合,并以100~120r/min进行搅拌10~15min,得混合物,按质量比1:8~10,先将步骤(1)中备用的浸泡后的银杏粉放入发酵罐的底部,在浸泡后得银杏粉上面放一层滤网,再将混合物倒入发酵罐中,在温度28~32℃下发酵10~15min,去除滤网,过滤,得发酵物;
(4)按质量比1:3~4,将发酵物、水进行混合,浸泡1~2h,再在微波频率为800~900MHz进行微波提取,得提取液,向提取液中加入发酵物质量10~20%的活性炭,30~40min后过滤,得滤液a,将滤液a在压力0.01~0.03MPa下减压浓缩,浓缩至滤液体积80~90%,再进行冷冻干燥,得干燥物,将干燥物进行粉碎,即完成银杏粉脱毒的方法。
实例1
处理液是按质量比1:1:40,将柠檬酸溶液、盐酸溶液、水进行混合,得混合物,即得处理液。
液体培养基是按重量份数计,取40份水、15份葡萄糖、5份蛋白胨、2份酵母浸粉、1份磷酸氢二钾、0.5份硫酸镁进行混合,高温灭菌,即得液体培养基。
PDA培养基是按重量份数计,取70份水、20份马铃薯 、2份葡萄糖 、1份琼脂进行混合,高温灭菌,即得PDA培养基。
种子培养基是按重量份数计,取50份水、15份琼脂、10份酵母膏、8份葡萄糖、5份蛋白胨、2份氯化钠进行混合,高温灭菌,即得种子培养基。
种子发酵培养基是按重量份数计,取70份水、50份柠檬酸氢二铵、30份可溶性淀粉、30份蔗糖、8份磷酸氢二铵、0.5份氯化钙、0.5份硫酸铁、0.5份硫酸镁进行混合,高温灭菌,即得种子培养基。
一种银杏粉脱毒的方法,该制备方法包括如下步骤:
(1)将银杏粉放入高压容器中处理,在常温下升压至8MPa,并以300r/min进行搅拌30min,1min内降压至常压,得处理后的银杏粉,按质量比1:3,将处理后的银杏粉、处理液进行混合,浸泡1h,得浸泡后的银杏粉,备用;
(2)按质量比1:3,将米糠、液体培养基在温度23℃下振荡培养5d,得培养液,将培养液用接种环划线到PDA培养基上,在温度28℃的温度下培养1d,用灭菌的接种环挑取PDA培养基的单菌落接种于种子培养基中,以100r/min摇床振荡扩大培养2d,挑取菌径最大的菌落,并接种于种子发酵培养基在温度28℃摇床中100r/min振荡培养30h,得培养液a,将培养液a用四层纱布过滤,得滤液,将滤液以1500r/min下离心10min,得上清液;
(3)按重量份数计,取10份上清液、7份百合、3份山楂核、3份燕麦进行混合,并以100r/min进行搅拌10min,得混合物,按质量比1:8,先将步骤(1)中备用的浸泡后的银杏粉放入发酵罐的底部,在浸泡后得银杏粉上面放一层滤网,再将混合物倒入发酵罐中,在温度28℃下发酵10min,去除滤网,过滤,得发酵物;
(4)按质量比1:3,将发酵物、水进行混合,浸泡1h,再在微波频率为800MHz进行微波提取,得提取液,向提取液中加入发酵物质量10%的活性炭,30min后过滤,得滤液a,将滤液a在压力0.01MPa下减压浓缩,浓缩至滤液体积80%,再进行冷冻干燥,得干燥物,将干燥物进行粉碎,即完成银杏粉脱毒的方法。
实例2
处理液是按质量比1:1:50,将柠檬酸溶液、盐酸溶液、水进行混合,得混合物,即得处理液。
液体培养基是按重量份数计,取50份水、20份葡萄糖、8份蛋白胨、4份酵母浸粉、2份磷酸氢二钾、1份硫酸镁进行混合,高温灭菌,即得液体培养基。
PDA培养基是按重量份数计,取80份水、30份马铃薯 、3份葡萄糖 、2份琼脂进行混合,高温灭菌,即得PDA培养基。
种子培养基是按重量份数计,取60份水、20份琼脂、15份酵母膏、10份葡萄糖、7份蛋白胨、4份氯化钠进行混合,高温灭菌,即得种子培养基。
种子发酵培养基是按重量份数计,取80份水、60份柠檬酸氢二铵、40份可溶性淀粉、40份蔗糖、10份磷酸氢二铵、1份氯化钙、1份硫酸铁、1份硫酸镁进行混合,高温灭菌,即得种子培养基。
一种银杏粉脱毒的方法,该制备方法包括如下步骤:
(1)将银杏粉放入高压容器中处理,在常温下升压至10MPa,并以400r/min进行搅拌40min,2min内降压至常压,得处理后的银杏粉,按质量比1:4,将处理后的银杏粉、处理液进行混合,浸泡2h,得浸泡后的银杏粉,备用;
(2)按质量比1:4,将米糠、液体培养基在温度28℃下振荡培养6d,得培养液,将培养液用接种环划线到PDA培养基上,在温度30℃的温度下培养2d,用灭菌的接种环挑取PDA培养基的单菌落接种于种子培养基中,以120r/min摇床振荡扩大培养3d,挑取菌径最大的菌落,并接种于种子发酵培养基在温度30℃摇床中120r/min振荡培养36h,得培养液a,将培养液a用四层纱布过滤,得滤液,将滤液以2000r/min下离心20min,得上清液;
(3)按重量份数计,取15份上清液、8份百合、5份山楂核、4份燕麦进行混合,并以120r/min进行搅拌15min,得混合物,按质量比1:10,先将步骤(1)中备用的浸泡后的银杏粉放入发酵罐的底部,在浸泡后得银杏粉上面放一层滤网,再将混合物倒入发酵罐中,在温度32℃下发酵15min,去除滤网,过滤,得发酵物;
(4)按质量比1:4,将发酵物、水进行混合,浸泡2h,再在微波频率为900MHz进行微波提取,得提取液,向提取液中加入发酵物质量20%的活性炭,40min后过滤,得滤液a,将滤液a在压力0.03MPa下减压浓缩,浓缩至滤液体积90%,再进行冷冻干燥,得干燥物,将干燥物进行粉碎,即完成银杏粉脱毒的方法。
实例3
处理液是按质量比1:1:45,将柠檬酸溶液、盐酸溶液、水进行混合,得混合物,即得处理液。
液体培养基是按重量份数计,取45份水、17.5份葡萄糖、6.5份蛋白胨、3份酵母浸粉、1.5份磷酸氢二钾、0.75份硫酸镁进行混合,高温灭菌,即得液体培养基。
PDA培养基是按重量份数计,取75份水、25份马铃薯 、2.5份葡萄糖 、1.5份琼脂进行混合,高温灭菌,即得PDA培养基。
种子培养基是按重量份数计,取55份水、17.5份琼脂、12.5份酵母膏、9份葡萄糖、6份蛋白胨、3份氯化钠进行混合,高温灭菌,即得种子培养基。
种子发酵培养基是按重量份数计,取75份水、55份柠檬酸氢二铵、35份可溶性淀粉、35份蔗糖、9份磷酸氢二铵、0.75份氯化钙、0.75份硫酸铁、0.75份硫酸镁进行混合,高温灭菌,即得种子培养基。
一种银杏粉脱毒的方法,该制备方法包括如下步骤:
(1)将银杏粉放入高压容器中处理,在常温下升压至9MPa,并以350r/min进行搅拌35min,1.5min内降压至常压,得处理后的银杏粉,按质量比1:3.5,将处理后的银杏粉、处理液进行混合,浸泡1.5h,得浸泡后的银杏粉,备用;
(2)按质量比1:3.5,将米糠、液体培养基在温度26℃下振荡培养5.5d,得培养液,将培养液用接种环划线到PDA培养基上,在温度29℃的温度下培养1.5d,用灭菌的接种环挑取PDA培养基的单菌落接种于种子培养基中,以110r/min摇床振荡扩大培养2.5d,挑取菌径最大的菌落,并接种于种子发酵培养基在温度29℃摇床中110r/min振荡培养33h,得培养液a,将培养液a用四层纱布过滤,得滤液,将滤液以1750r/min下离心15min,得上清液;
(3)按重量份数计,取12.5份上清液、7.5份百合、4份山楂核、3.5份燕麦进行混合,并以110r/min进行搅拌12.5min,得混合物,按质量比1:9,先将步骤(1)中备用的浸泡后的银杏粉放入发酵罐的底部,在浸泡后得银杏粉上面放一层滤网,再将混合物倒入发酵罐中,在温度30℃下发酵12.5min,去除滤网,过滤,得发酵物;
(4)按质量比1:3.5,将发酵物、水进行混合,浸泡1.5h,再在微波频率为850MHz进行微波提取,得提取液,向提取液中加入发酵物质量15%的活性炭,35min后过滤,得滤液a,将滤液a在压力0.02MPa下减压浓缩,浓缩至滤液体积85%,再进行冷冻干燥,得干燥物,将干燥物进行粉碎,即完成银杏粉脱毒的方法。
对照例:传统银杏粉脱毒方法。
分别使用本发明的脱毒方法与对照例的脱毒方法对银杏粉进行脱毒处理,采用高效薄层色谱方法测试其中物质含量以此计算毒素的去除率与活性物质的损失率,测试结果见表1。
测试项目 | 实例1 | 实例2 | 实例3 | 对照例 |
脱毒时间h | 4 | 3 | 3 | 12~15 |
总毒素去除率% | 98 | 99 | 98 | 61~66 |
总活性物质损失率% | 7 | 5 | 6 | 43~52 |
安全性测试:将小白鼠分别以0.1mg/10g、0.2mg/10g、0.4mg/10g经过本发明脱毒后的银杏粉灌胃,每天灌胃一次,连续观察3周,观察灌胃后小白鼠情况。结果:21天内未见小白鼠有中枢兴奋或抑制症状,未见肌张力改变或肌震颤症状,小白鼠活动自如步态正常,未出现呼吸异常,进食,二便正常,观察期间小白鼠无一例死亡。
综合上述,本发明的银杏粉脱毒方法脱毒效果好,毒素基本无残留,值得推广。
Claims (10)
1.一种银杏粉脱毒的方法,其特征在于,该制备方法包括如下步骤:
(1)将银杏粉放入高压容器中处理,在常温下升压至8~10MPa,进行搅拌30~40min,1~2min内降压至常压,得处理后的银杏粉,将处理后的银杏粉、处理液进行混合,浸泡1~2h,得浸泡后的银杏粉,备用;
(2)将米糠、液体培养基在温度23~28℃下振荡培养5~6d,得培养液,将培养液用接种环划线到PDA培养基上,在温度28~30℃的温度下培养1~2d,用灭菌的接种环挑取PDA培养基的单菌落接种于种子培养基中摇床振荡扩大培养2~3d,挑取菌径最大的菌落,并接种于种子发酵培养基在温度28~30℃摇床中振荡培养30~36h,得培养液a,将培养液a用四层纱布过滤,得滤液,将滤液进行离心10~20min,得上清液;
(3)按重量份数计,取10~15份上清液、7~8份百合、3~5份山楂核、3~4份燕麦进行混合,并进行搅拌10~15min,得混合物,先将步骤(1)中备用的浸泡后的银杏粉放入发酵罐的底部,在浸泡后得银杏粉上面放一层滤网,再将混合物倒入发酵罐中,在温度28~32℃下发酵10~15min,去除滤网,过滤,得发酵物;
(4)将发酵物、水进行混合,浸泡1~2h,再进行微波提取,得提取液,向提取液中加入发酵物质量10~20%的活性炭,30~40min后过滤,得滤液a,将滤液a进行减压浓缩,浓缩至滤液体积80~90%,再进行冷冻干燥,得干燥物,将干燥物进行粉碎,即完成银杏粉脱毒方法。
2.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(1)中处理后的银杏粉、处理液的质量比1:5~6。
3.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(1)中处理液是按质量比1:1:40~50,将柠檬酸溶液、盐酸溶液、水进行混合,得混合物,即得处理液。
4.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(2)中米糠、液体培养基的质量比1:7~8。
5.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(2)中液体培养基是按重量份数计,取40~50份水、15~20份葡萄糖、5~8份蛋白胨、2~4份酵母浸粉、1~2份磷酸氢二钾、0.5~1份硫酸镁进行混合,高温灭菌,即得液体培养基。
6.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(2)中PDA培养基是按重量份数计,取70~80份水、20~30份马铃薯 、2~3份葡萄糖 、 1~2份琼脂进行混合,高温灭菌,即得PDA培养基。
7.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(2)中种子培养基是按重量份数计,取50~60份水、15~20份琼脂、10~15份酵母膏、8~10份葡萄糖、5~7份蛋白胨、2~4份氯化钠进行混合,高温灭菌,即得种子培养基。
8.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(2)中种子发酵培养基是按重量份数计,取70~80份水、50~60份柠檬酸氢二铵、30~40份可溶性淀粉、30~40份蔗糖、8~10份磷酸氢二铵、0.5~1份氯化钙、0.5~1份硫酸铁、0.5~1份硫酸镁进行混合,高温灭菌,即得种子培养基。
9.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(3)中步骤(1)中备用的浸泡后的银杏粉、混合物的质量比1:8~10。
10.根据权利要求1所述银杏粉脱毒的方法,其特征在于,所述步骤(4)中发酵物、水的质量比1:3~4。
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