CN107459547B - 蛋黄中多种生物活性物质的联产分离方法 - Google Patents
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Abstract
本发明公开一种蛋黄中多种生物活性物质的联产分离方法,该方法步骤如下:(1)胆固醇的提取,(2)卵磷脂的提取;(3)卵黄高磷蛋白的提取。本发明先利用超临界CO2萃取法提取出蛋黄油,之后用醇提法萃取卵磷脂,一方面提高了胆固醇的提取率,一方面提高了蛋黄卵磷脂的纯度。本发明进行卵磷脂提取时,以无毒的食用乙醇为溶媒,提高了产品食用安全性,采用酶解辅助提取,有利于脂质与蛋白之间的共价键断裂,加快了蛋黄卵磷脂的提取率,缩短了生产周期。采用超滤技术对卵黄高磷蛋白进行脱盐,进一步提高了产品纯度,且工艺简单,操作方便,适合于大规模工业化生产。
Description
技术领域
本发明属于食品加工和提取技术领域,特别涉及蛋黄中多种生物活性物质的联产分离方法。
背景技术
我国是禽蛋生产第一大国,以鸡蛋为主,约占84%,其余为鸭蛋及鹅蛋。目前消费主要以鲜蛋为主,用于深加工的只有0.7%~1%。蛋黄作为禽蛋中营养价值最高的一部分,其总量约占禽蛋总重的28%~29%,含有大量的功能性成分,均具有明显的生物活性。脂质是蛋黄干物质中含量最为丰富的成分,约占干物质的三分之二,主要有甘油三酸酯、卵磷脂、胆固醇等。除了脂质以外,蛋黄中另一种主要成分就是蛋黄蛋白质,大部分以脂蛋白的形式存在,包括65%的低密度脂蛋白、16%的高密度脂蛋白、10%的卵黄球蛋白和4%的卵黄高磷蛋白,其中低密度脂蛋白极易被蛋白酶分解而释放出蛋黄卵磷脂。这些活性物质具有独特的理化性质和生理功能,被广泛应用于食品、保健品、化妆品、医药和饲料工业等领域。
目前对蛋黄的研究主要集中于某一种生物活性成分的提取及精制方面,但由于现有技术水平落后,生产装置规模小,导致分离所得的单一活性成分产量少、纯度也不高,远远不能满足现有市场的需求,同时,对两种或两种以上高附加值天然产物的联产技术的研究更是鲜有报道,并没有实现真正意义上的综合利用。本发明期望实现胆固醇、卵磷脂、卵黄高磷蛋白等成分的联产,建立一套产率高、纯度高、效益高、可控性好的蛋品精深加工技术,在有效提高蛋品综合利用水平的同时促进禽蛋养殖业的持续稳定发展及保健产品的开发。
发明内容
本发明针对现有蛋黄生物活性物质提取及联产方面存在的缺陷及不足,提供一种胆固醇、卵磷脂及卵黄高磷蛋白的联产方法,用于提高蛋黄中生物活性物质提取的产量及纯度,同时增加禽蛋精深加工的综合利用率,真正实现禽蛋的高值化利用。
本发明采用的技术方案为:一种蛋黄中多种生物活性物质的联产分离方法,该方法步骤如下:
(1)胆固醇的提取:
步骤(1)挑选新鲜禽蛋,洗净、破壳后分离出蛋黄,将蛋黄在45~50℃下干燥至水分含量<5%,粉碎获得蛋黄粉,采用超临界CO2萃取法提取蛋黄油,剩余物料备用;
步骤(2)蛋黄油中加入体积比为1:2~6的质量浓度8%β-环糊精-壳聚糖水溶液,在20~45℃下搅拌20~45min,离心后采用热乙醇浸提法从胆固醇-β-环糊精-壳聚糖复合物中分离出胆固醇;
(2)卵磷脂的提取:以步骤(1)的剩余物料为原料,加入3~12倍体积的80~90%食用乙醇,搅拌均匀后加入0.1~0.4%的提取蛋白酶,在40~50℃下酶解1~2h,离心后获得上层液及蛋黄蛋白,上层液中主要成分是蛋黄卵磷脂,将其在40~48℃下真空浓缩至稠状,得到纯度为87.2%的卵磷脂粗品,使用0℃的乙酸乙酯洗涤卵磷脂粗品后在40~50℃下干燥,蛋黄卵磷脂的纯度提高到90%以上;
(3)卵黄高磷蛋白的提取:在(2)所述的蛋黄蛋白中加入体积比为1:2~10的质量浓度10%NaCl溶液,调节pH为3.0~6.0,搅拌0.5~2h,离心后获得滤液,将滤液经过截留分子量为10kDa的超滤膜装置进行脱盐,脱去多余的盐分后冷冻干燥获得卵黄高磷蛋白。
本发明的方法进一步设置为,所述步骤(1)中蛋黄油萃取条件为萃取釜温度为30℃-46℃,压力为18MPa-38MPa,分离釜温度为40℃-50℃,压力为4MPa-8MPa,连续萃取40~120min。收集蛋黄油。
本发明的方法进一步设置为,所述β-环糊精与壳聚糖水溶液采用质量浓度8%的β-环糊精-壳聚糖水溶液包埋胆固醇,其中β-环糊精与壳聚糖按1~4:1的体积比例混合,包埋温度为20~45℃。
本发明的方法进一步设置为,所述步骤(1)中胆固醇的分离方法具体为:将胆固醇-β-环糊精复合物沉淀以1:2~10的比例加入浓度为95~100%的乙醇,在50~70℃搅拌0.5~2h,之后将热乙醇提取液在0~4℃下静置5~10h,析出胆固醇,经过滤、干燥获得精制胆固醇;
本发明的方法进一步设置为,所述步骤(2)中的提取蛋白酶由酸性蛋白酶:木瓜蛋白酶=3:1制得。
本发明的优点在于:
(1)本发明采用超临界CO2萃取法、微胶囊技术、酶解辅助有机溶剂萃取法、盐析分离技术,同时获得了高纯度、高品质的胆固醇、卵磷脂及卵黄高磷蛋白3个产品,提高了资源的利用率,实现了蛋黄的高值化全利用。
(2)本发明先利用超临界CO2萃取法提取出蛋黄油,之后用醇提法萃取卵磷脂,一方面提高了胆固醇的提取率,一方面提高了蛋黄卵磷脂的纯度。
(3)本发明进行卵磷脂提取时,以无毒的食用乙醇为溶媒,提高了产品食用安全性,采用酶解辅助提取,有利于脂质与蛋白之间的共价键断裂,加快了蛋黄卵磷脂的提取率,缩短了生产周期。
(4)β-环糊精与壳聚糖的混合物对胆固醇的吸附能力要显著高于单独使用β-环糊精,将两者混合后从蛋黄油中分离出胆固醇,显著提高了胆固醇的回收率。
(5)采用超滤技术对卵黄高磷蛋白进行脱盐,进一步提高了产品纯度,且工艺简单,操作方便,适合于大规模工业化生产。
附图说明
图1本发明的流程图。
具体实施方式
下面结合具体实施例,进一步阐述本发明。
在不背离本发明精神和实质的情况下,对本发明的方法、步骤或条件所做的修改或替换,均属于本发明范围,若未特别指明,实施例中所用的技术手段为本领域技术人员所熟悉的常规手段。
实施例1
一种蛋黄中多种生物活性物质的联产分离方法,该方法步骤如下:
(1)胆固醇的提取:
a挑选新鲜禽蛋,洗净、破壳后分离出蛋黄,将蛋黄在45~50℃下干燥至水分含量<5%,粉碎获得蛋黄粉,采用超临界CO2萃取法提取蛋黄油,剩余物料备用;蛋黄油萃取条件为萃取釜温度为30℃-46℃,压力为18MPa-38MPa,分离釜温度为40℃-50℃,压力为4MPa-8MPa,连续萃取40~120min。收集蛋黄油。胆固醇提取方法中使用超临界CO2萃取法可以提取出将近99%的胆固醇及甘油三酯,提高了胆固醇的提取率,同时也大大减少了后续蛋黄卵磷脂提取的杂质,有效提高了其纯度。
b蛋黄油中加入体积比为1:2~6的质量浓度8%β-环糊精-壳聚糖水溶液,在20~45℃下搅拌20~45min,离心后采用热乙醇浸提法从胆固醇-β-环糊精-壳聚糖复合物中分离出胆固醇;所述β-环糊精与壳聚糖水溶液采用8%的β-环糊精-壳聚糖水溶液包埋胆固醇,其中β-环糊精与壳聚糖按1~4:1的体积比例混合,包埋温度为20~45℃。所述步骤(1)中胆固醇的分离方法具体为:将胆固醇-β-环糊精复合物沉淀以1:2~10的比例加入浓度为95~100%的乙醇,在50~70℃搅拌0.5~2h,之后将热乙醇提取液在0~4℃下静置5~10h,析出胆固醇,经过滤、干燥获得精制胆固醇.同时,利用β-环糊精及壳聚糖对胆固醇的特殊吸附能力,将两者混合后从蛋黄油中分离出胆固醇,显著提高了胆固醇的回收率。
(2)卵磷脂的提取:以步骤(1)的剩余物料为原料,加入3~12倍体积的80~90%食用乙醇,搅拌均匀后加入0.1~0.4%的提取蛋白酶,在40~50℃下酶解1~2h,离心后获得上层液及蛋黄蛋白,上层液中主要成分是蛋黄卵磷脂,将其在40~48℃下真空浓缩至稠状,得到纯度为87.2%的卵磷脂粗品,使用0℃的乙酸乙酯洗涤卵磷脂粗品后在40~50℃下干燥,蛋黄卵磷脂的纯度提高到90%以上;所述步骤(2)中的提取蛋白酶由酸性蛋白酶:木瓜蛋白酶=3:1制得。利用酶解与有机溶剂萃取法同步进行来提取脱油蛋黄渣中的卵磷脂,加快了脂质与蛋白之间的共价键的断裂及蛋黄卵磷脂的溶出速率,提高了产量,缩短了生产周期。利用乙酸乙酯作为蛋黄卵磷脂的洗涤剂,有效提高了卵磷脂的纯度。
(3)卵黄高磷蛋白的提取:在步骤(2)所述的蛋黄蛋白中加入体积比为1:2~10的质量浓度10%NaCl溶液,调节pH为3.0~6.0,搅拌0.5~2h,离心后获得滤液,将滤液经过截留分子量为10kDa的超滤膜装置进行脱盐,脱去多余的盐分后冷冻干燥获得卵黄高磷蛋白。
以上仅是本发明的优选实施方式,本发明的保护范围并不仅局限于实施例,凡属于本发明思路下的技术方案均属于本发明的保护范围。应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理前提下的若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (1)
1.蛋黄中多种生物活性物质的联产分离方法,其特征在于,所述多种生物活性物质包括胆固醇、卵磷脂、卵黄高磷蛋白,所述方法步骤如下:
步骤(1)胆固醇的提取:
a. 挑选新鲜禽蛋,洗净、破壳后分离出蛋黄,将蛋黄在45~50℃下干燥至水分含量<5%,粉碎获得蛋黄粉,采用超临界CO2萃取法提取蛋黄油,剩余物料备用;
b. 蛋黄油中加入体积比为1:2~6的8%β-环糊精-壳聚糖水溶液,在20~45℃下搅拌20~45min,离心后采用热乙醇浸提法从胆固醇-β-环糊精-壳聚糖复合物中分离出胆固醇;
采用超临界CO2萃取法提取出99%的胆固醇及甘油三酯;
步骤(2)卵磷脂的提取:以步骤(1)的所述剩余物料为原料,加入3~12倍体积的80~90%食用乙醇,搅拌均匀后加入0.1~0.4%的提取蛋白酶,在40~50℃下酶解1~2h,离心后获得上层液及蛋黄蛋白,上层液中主要成分是蛋黄卵磷脂,将其在40~48℃下真空浓缩至稠状,得到纯度为87.2%的卵磷脂粗品,使用0℃的乙酸乙酯洗涤卵磷脂粗品后在40~50℃下干燥,蛋黄卵磷脂的纯度提高到90%以上;
(3)卵黄高磷蛋白的提取:在步骤(2)所述的蛋黄蛋白中加入体积比为1:2~10的10%NaCl溶液,调节pH为3.0~6.0,搅拌0.5~2h,离心后获得滤液,将滤液经过截留分子量为10kDa的超滤膜装置进行脱盐,脱去多余的盐分后冷冻干燥获得卵黄高磷蛋白;
所述步骤(1)中蛋黄油萃取条件为萃取釜温度为30℃~46℃,压力为18MPa~38MPa,分离釜温度为40℃~50℃,压力为4MPa~8MPa,连续萃取40~120min,收集蛋黄油;
所述β-环糊精与壳聚糖水溶液采用8%的β-环糊精-壳聚糖水溶液包埋胆固醇,其中β-环糊精与壳聚糖按1~4:1的体积比例混合,包埋温度为20~45℃;
所述步骤(2)中的提取蛋白酶由酸性蛋白酶:木瓜蛋白酶=3:1制得;所述步骤(1)中胆固醇的分离方法具体为:将胆固醇-β-环糊精复合物沉淀以1:2~10的比例加入浓度为95~100%的乙醇,在50~70℃搅拌0.5~2h,之后将热乙醇提取液在0~4℃下静置5~10h,析出胆固醇,经过滤、干燥获得精制胆固醇。
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