CN108835490A - 一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法 - Google Patents
一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法 Download PDFInfo
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Abstract
本发明公开了一种稳定的大黄鱼卵分离蛋白‑β‑胡萝卜素乳液的制备方法,包括如下步骤:S1、制备蛋白冻干粉:用氯化钠溶液对大黄鱼卵进行盐溶蛋白提取,离心得到上清液,冻干备用;S2、制备油相:将β‑胡萝卜素通过超声加热溶解于玉米油中,配制成油相待用;S3、制备水相:用缓冲液配制一定浓度的蛋白溶液,配制成水相待用;S4、混合:将油相与水相按一定比例混合,高速分散;S5、均质:高压均质后得到稳定的大黄鱼卵分离蛋白‑β‑胡萝卜素乳液。本发明利用大黄鱼卵分离蛋白作为乳化剂,包埋β‑胡萝卜素得到的大黄鱼卵分离蛋白‑β‑胡萝卜素产物乳液,营养价值大,稳定性高。
Description
技术领域
本发明涉及乳液制备技术领域,具体地说,涉及一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法。
背景技术
脂溶性营养物质是指一类能够溶解在脂肪中,进而被人体摄入、消化、吸收的物质。常见的脂溶性营养物质包括维生素类、类胡萝卜素类、黄酮类、多酚类、多不饱和脂肪酸类等。为了提高脂溶性营养物质在人体内的生物利用率,常见的方法是利用水包油乳液体系对其进行包埋。
将脂溶性营养物质作为芯材进行传递时,可以有多种保护其不被降解的方法,常用蛋白作为乳化剂加以稳定。蛋白在油水界面上通过降低界面张力以及增大空间位阻起到稳定作用。目前多见以植物蛋白作为乳化剂,对其构建的乳液系统稳定性的研究,对于水产来源蛋白构筑的乳液系统的研究较少。
大黄鱼(Pseudosciaena crocea),又名大王鱼、石头鱼等,广泛分布于我国东、南海及黄海南部,是我国传统“四大海产”之一。大黄鱼鱼卵是大黄鱼加工的主要剩余物。经实测,鱼卵大约占鱼体总质量的15%~25%,其营养价值极高,除富含多种长链不饱和脂肪酸和磷脂,还含有丰富的蛋白质,如茴鱼卵、史氏鲟鱼卵和鳟鱼卵等鲜鱼卵中的蛋白质含量均超过30%,但都得不到良好的利用。目前,除少部分鱼类的鱼卵,如鲑鱼、鲟鱼和鳟鱼等鱼卵被加工成美味且价格昂贵的鱼籽酱,少量的鱼卵被加工成饲料外,其余均被当作废弃物丢弃,造成了极大的资源浪费和环境污染。在我国,大黄鱼卵的开发利用及相关研究目前鲜有报道。因此,如何高效合理的利用大黄鱼卵得到了越来越多的关注。
β-胡萝卜素是自然界中分布最广、也是目前学者研究最多的类胡萝卜素,在食品工业和食品科学研究中非常重要。β-胡萝卜素具有多种生物学功能,其分子中有两个β-紫罗酮环,具有100%的维生素A前体活性。此外,其潜在的猝灭单线态氧和使自由基失活的活性在预防特定的癌症、心血管疾病、黄斑退化症、白内障和提高免疫反应中也发挥了较大作用。因此,β-胡萝卜素是一种对人体健康非常重要的生物活性物质。
β-胡萝卜素通常会在光、热、酶、金属离子、与金属结合的蛋白质和微生物的作用下加速降解,引起一系列的氧化反应,导致生物活性降低,颜色退化,产生不良风味和潜在的有毒物质,生物利用率下降等。因此,利用水包油乳液传递β-胡萝卜素是一种成本较低,且可以提高其水溶性、稳定性及生物利用率的方法,在食品工业具有较好的应用前景。
发明内容
本发明针对鱼卵来源蛋白乳液体系,提供一种稳定的大黄鱼卵分离蛋白-β-胡萝卜素乳液。
为了达到上述目的,本发明提供了一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,包括以下步骤:
S1、制备蛋白冻干粉:用氯化钠溶液对大黄鱼卵进行蛋白提取,离心,取上清液(即蛋白提取液)冻干,得蛋白冻干粉备用;
S2、制备油相:将β-胡萝卜素超声加热,使其溶解于玉米油中,配制成油相待用;
S3、制备水相:将步骤S1所得蛋白冻干粉用磷酸盐缓冲液(PBS)复溶,配制成水相待用;
S4、混合:将步骤S2配制的油相与步骤S3配制的水相混合,高速分散;
S5、均质:将步骤S4所得产物均质,获得产物乳液。
优选方式下,步骤S1所述制备蛋白冻干粉具体为:用浓度0.1~1mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵和氯化钠溶液的料液比为(1:10)~(1:20)g/mL,5000~10000rpm/min离心10~30min,取上清液(即蛋白提取液),-25~-50℃条件下真空冷冻干燥60~72h,得蛋白冻干粉备用;
优选方式下,步骤S2所述制备油相具体为:将β-胡萝卜素在10~30kHz、50~55℃条件下超声加热10~20min,溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.05%~0.1%的油相待用;
优选方式下,步骤S3所述制备水相具体为:将步骤S1所得蛋白冻干粉用pH7~9,5~15mmol/L的PBS复溶,配制成蛋白浓度为2~5mg/mL的水相待用;
优选方式下,步骤S4所述混合具体为:将步骤S2配制的油相与步骤S3配制的水相按重量比(2~5):100的比例混合,10000~15000r/min高速分散2~3min;
优选方式下,步骤S5所述均质具体为:将步骤S4制备的产物10000~12000psi高压均质3~7次,得产物乳液。
本发明的有益效果是:
本发明涉及一种稳定的大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,包括步骤:S1、用氯化钠溶液对大黄鱼卵进行盐溶蛋白提取,离心得到上清液,冻干备用。S2、将β-胡萝卜素通过超声加热溶解于玉米油中,配制成油相待用;S3、用缓冲液配制一定浓度的蛋白溶液,配制成水相待用;S4、将油相与水相按一定比例混合,高速分散;S5、高压均质后得到稳定的大黄鱼卵分离蛋白-β-胡萝卜素乳液。本发明利用大黄鱼卵分离蛋白作为乳化剂,包埋β-胡萝卜素得到一种稳定且具有一定营养价值的大黄鱼卵分离蛋白-β-胡萝卜素乳液。
本发明采用鱼卵蛋白作为乳化剂,在提升乳液体系稳定性的同时也提高了产品的营养价值,制备方法简单易行,制作出的乳液具有较好的稳定性,可用于进一步制备保健品或富含β-胡萝卜素的功能性饮品,作为一种人体补充β-胡萝卜素的新方式,具有一定的市场价值。
附图说明
图1是本发明实施例1~实施例4制备的产物乳液在4℃存放1天的粒径分布图;
图2是本发明实施例1~实施例4制备的产物乳液在常温存放1天的粒径分布图;
图3是本发明实施例1~实施例4制备的产物乳液在4℃存放4天的粒径分布图;
图4是本发明实施例1~实施例4制备的产物乳液在常温存放4天的粒径分布图;
图5是本发明实施例1~实施例4制备的产物乳液在4℃存放7天的粒径分布图;
图6是本发明实施例1~实施例4制备的产物乳液在常温存放7天的粒径分布图;
图7是本发明实施例1~实施例4制备的产物乳液在4℃存放14天的粒径分布图;
图8是本发明实施例1~实施例4制备的产物乳液在常温存放14天的粒径分布图;
图9是本发明实施例1~实施例4制备的产物乳液在4℃存放1天、4天、7天及14天的平均粒径;
图10是本发明实施例1~实施例4制备的产物乳液在常温存放1天、4天、7天及14天的平均粒径;
图11是本发明实施例1~实施例4制备的产物乳液在4℃存放1天、4天、7天及14天的ζ-电位图;
图12是本发明实施例1~实施例4制备的产物乳液在常温存放1天、4天、7天及14天的ζ-电位图;
图13是本发明实施例1制备的产物乳液在4℃存放0天、1天、4天、7天及14天的β-胡萝卜素保留率,对照为无大黄鱼卵蛋白包埋的β-胡萝卜素油相溶液;
图14是本发明实施例1制备的产物乳液在常温存放0天、1天、4天、7天及14天的β-胡萝卜素保留率,对照为无大黄鱼卵蛋白包埋的β-胡萝卜素油相溶液;
具体实施方式
本发明提供一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,取大黄鱼卵分离蛋白和β-胡萝卜素,制备步骤如下:
S1、用浓度0.1~1mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,料液比(1:10)~(1:20)g/mL,5000~10000rpm/min离心10~30min,取上清液(即蛋白提取液),-25~-50℃条件下真空冷冻干燥60~72h,得蛋白冻干粉备用;
S2、将β-胡萝卜素10~30kHz、50~55℃超声加热10~20min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.05%~0.1%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH7~9,5~15mmol/L的PBS复溶,配制成蛋白浓度为2~5mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量(2~5):100的比例混合,10000~15000r/min高速分散2~3min;
S5、10000~12000psi高压均质3~7次,得产物乳液。
实施例1
一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,取大黄鱼卵分离蛋白和β-胡萝卜素,其制备步骤如下:
S1、用浓度0.6mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵和氯化钠溶液的料液比1:20g/mL,5000rpm/min离心30min,取上清液(即蛋白提取液),-25℃条件下真空冷冻干燥60h,得蛋白冻干粉备用;
S2、将β-胡萝卜素10kHz、50℃超声加热20min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.1%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH 8,10mmol/L的PBS复溶,配制成蛋白浓度为2mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量2:100的比例混合,10000r/min高速分散2min;
S5、12000psi高压均质5次,得产物乳液。
实施例2
S1、用浓度0.6mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,料液比1:20g/mL,5000rpm/min离心30min,取上清液(即蛋白提取液),-50℃条件下真空冷冻干燥72h,得蛋白冻干粉备用;
S2、将β-胡萝卜素10kHz、50℃超声加热20min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.1%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH 8,10mmol/L的PBS复溶,配制成蛋白浓度为2mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量3:100的比例混合,10000r/min高速分散2min;
S5、12000psi高压均质5次,得产物乳液。
实施例3
S1、用浓度0.6mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵和氯化钠溶液的料液比为1:20g/mL,5000rpm/min离心30min,取上清液(即蛋白提取液),-30℃条件下真空冷冻干燥65h,得蛋白冻干粉备用;
S2、将β-胡萝卜素10kHz、50℃超声加热20min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.1%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH 8,10mmol/L的PBS复溶,配制成蛋白浓度为2mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量4:100的比例混合,10000r/min高速分散2min;
S5、12000psi高压均质5次,得产物乳液。
实施例4
S1、用浓度0.6mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵和氯化钠溶液的料液比为1:20g/mL,5000rpm/min离心30min,取上清液(即蛋白提取液),-40℃条件下真空冷冻干燥70h,得蛋白冻干粉备用;
S2、将β-胡萝卜素10kHz、50℃超声加热20min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.1%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH 8,10mmol/L的PBS复溶,配制成蛋白浓度为2mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量5:100的比例混合,10000r/min高速分散2min;
S5、12000psi高压均质5次,得产物乳液。
实施例5
S1、用浓度0.1mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵和氯化钠溶液的料液比为1:15g/mL,10000rpm/min离心10min,取上清液(即蛋白提取液),-45℃条件下真空冷冻干燥70h,得蛋白冻干粉备用;
S2、将β-胡萝卜素30kHz、55℃超声加热10min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.05%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH 7,5mmol/L的PBS复溶,配制成蛋白浓度为5mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量5:100的比例混合,15000r/min高速分散3min;
S5、12000psi高压均质3次,得产物乳液。
实施例6
S1、用浓度1mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵和氯化钠溶液的料液比为1:10g/mL,8000rpm/min离心20min,取上清液(即蛋白提取液),-45℃条件下真空冷冻干燥70h,得蛋白冻干粉备用;
S2、将β-胡萝卜素20kHz、53℃超声加热15min,使其溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.08%的油相待用;
S3、将步骤S1所得蛋白冻干粉用pH 9,15mmol/L的PBS复溶,配制成蛋白浓度为3mg/mL的水相待用;
S4、将步骤S2配制的油相与步骤S3配制的水相按重量5:100的比例混合,12000r/min高速分散2.5min;
S5、10000psi高压均质7次,得产物乳液。
对本发明实施例制备的产物乳液的粒径和ζ-电位进行测定:取适量乳液(4℃和常温分别存放1天、4天、7天及14天)稀释100倍后,采用激光粒度仪进行平均粒径,粒径分布以及ζ-电位的检测。因为实施例1在常温存放14天后体系最稳定,故对其进行β-胡萝卜素保留率检测,取适量本发明实施例制备的乳液及未经大黄鱼卵蛋白包埋的质量浓度0.05%~0.1%的β-胡萝卜素油相溶液(4℃和常温分别存放0天、1天、4天、7天及14天)提取β-胡萝卜素后,采用酶标仪测定其在450nm下的吸光度,分析其保留率,数据中不同字母代表显著性差异(p<0.05)。具体分析及理论依据包括:
(1)乳液平均粒径越小,体系越稳定。
(2)ζ-电位绝对值越高,体系越稳定。
(3)样品中β-胡萝卜素含量与其在450nm下的吸光度成线性关系。
检测结果如图1~图14所示,数据图中不同字母代表显著性差异(p<0.05),通过检测数据对比可知,本发明制备的产物乳液平均粒径小,ζ-电位绝对值高;经过14天相同条件下的等温储藏后,β-胡萝卜素保留率如下:常温条件,乳液样品85.58%±2.02%,对照76.94%±3.06%,具有显著性差异;4℃条件,乳液样品89.68%±2.06%,对照83.58%±3.16%,具有显著性差异。综上所述,乳液中的β-胡萝卜素保留率与对照相比具有显著的提高(所述对照为未经大黄鱼卵蛋白包埋的质量浓度为0.05%~0.1%的β-胡萝卜素油相溶液),即本发明所制得乳液具有较高的稳定性。
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 (6)
1.一种大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,包括以下步骤:
S1、制备蛋白冻干粉:用氯化钠溶液对大黄鱼卵进行蛋白提取,离心,取上清液,冻干得蛋白冻干粉备用;
S2、制备油相:将β-胡萝卜素溶解于玉米油中,配制成油相待用;
S3、制备水相:将步骤S1所得蛋白冻干粉用磷酸盐缓冲液配制成水相待用;
S4、混合:将步骤S2配制的油相与步骤S3配制的水相混合,高速分散;
S5、均质:将步骤S4所得产物均质。
2.根据权利要求1所述大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,其特征在于,步骤S1所述制备蛋白冻干粉具体为:用浓度0.1~1mol/L的氯化钠溶液对大黄鱼卵进行蛋白提取,大黄鱼卵与氯化钠溶液料液比(1:10)~(1:20)g/mL,5000~10000rpm/min离心10~30min,取上清液,-25~-50℃条件下真空冷冻干燥60~72h,得蛋白冻干粉备用。
3.根据权利要求1所述大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,其特征在于,步骤S2所述制备油相具体为:将β-胡萝卜素在10~30kHz、50~55℃条件下超声加热10~20min,使β-胡萝卜素溶解于玉米油中,配制成β-胡萝卜素质量浓度为0.05%~0.1%的油相待用。
4.根据权利要求1所述大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,其特征在于,步骤S3所述制备水相具体为:将步骤S1所得蛋白冻干粉用pH 7~9,5~15mmol/L的磷酸盐缓冲液复溶,配制成蛋白浓度为2~5mg/mL的水相待用。
5.根据权利要求1所述大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,其特征在于,步骤S4所述混合具体为:将步骤S2配制的油相与步骤S3配制的水相按重量比(2~5):100的比例混合,10000~15000r/min高速分散2~3min。
6.根据权利要求1所述大黄鱼卵分离蛋白-β-胡萝卜素乳液的制备方法,其特征在于,步骤S5所述均质具体为:10000~12000psi高压均质3~7次。
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CN111838395A (zh) * | 2020-08-13 | 2020-10-30 | 湖北欣和生物科技有限公司 | 一种鱼肉分离蛋白基的乳液冷凝胶的简易制备方法 |
CN115428935A (zh) * | 2022-06-30 | 2022-12-06 | 大连工业大学 | 一种新型大黄鱼卵分离蛋白/结冷胶复合凝胶的制备方法 |
CN115428935B (zh) * | 2022-06-30 | 2024-03-19 | 大连工业大学 | 一种大黄鱼卵分离蛋白/结冷胶复合凝胶的制备方法 |
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