CN114601170A - 一种溶解血红素的新方法 - Google Patents
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Abstract
本发明公开了一种溶解血红素的新方法,属于食品加工领域。本发明方法包括以下步骤:(1)制备大豆分离蛋白浓度为1.0~2.0%、pH为11.0~12.0的大豆分离蛋白溶液;(2)将血红素或血红素衍生化合物加入到大豆分离蛋白溶液中,调节pH至6.5~7.5,得到大豆分离蛋白‑血红素复合纳米颗粒溶液;(3)将大豆分离蛋白‑血红素复合纳米颗粒溶液冻干或喷雾干燥,得到可溶性血红素。本发明制备的可溶性血红素,几乎完全溶解于水,乳化能力强,可以作为补铁剂广泛应用于各类食品或医药中。
Description
技术领域
本发明属于食品加工领域,具体涉及一种溶解血红素的新方法。
背景技术
缺铁性贫血是全世界发病率最高的营养缺乏病之一,严重威胁着人类的健康,目前,缺铁性贫血的防治主要通过补铁剂实现,包括硫酸亚铁、氯化亚铁、葡萄糖酸亚铁、乳酸亚铁、琥珀酸亚铁、富马酸亚铁等,虽然这些补铁剂的铁含量高,但由于体内利用率低、毒副作用大,并且有特殊的金属铁锈味,难以长期食用。血红素铁是一种生物态铁,可以直接被肠粘膜细胞吸收,不产生任何消化道刺激症状,生物利用率高,是理想的补铁剂。
血红素(Heme)是生物体内的卟啉铁配合物,机体中的铁大约70%以血红素的形式存在,在生物体内发挥极其重要的生理功能。血红素在食品、医药工业中有非常重要的应用价值。在食品工业中,可添加到糖果、饼干、面条、面包、米粉、方便汤料奶制品、酱油等食品中,在肉制品中添加还可以起到发色剂的作用。在医药领域中,可用于治疗贫血病、辅助治疗铅中毒、制备胆红素等。血红素不溶于水,溶解于酸性丙酮和碱性水溶液,因此在很多食品中的应用受到限制。一些研究采用环糊精包埋血红素在一定程度上解决其难溶于水的问题,但包埋率比较低。
发明内容
本发明的目的在于克服现有技术中血红素难溶于水的问题,提供一种溶解血红素的新方法。本发明的目的还在于提供一种可溶性血红素。
本发明的目的通过下述技术方案实现:
一种溶解血红素的方法,包括以下步骤:
(1)制备大豆分离蛋白(SPI)浓度为1.0~2.0%(w/v)、pH为11.0~12.0的SPI溶液。
(2)将血红素或血红素衍生化合物(如氯化血红素)(统一简写为He)加入到SPI溶液中,完全溶解后,调节pH至6.5~7.5,得到SPI-He复合纳米颗粒溶液。
(3)将SPI-He复合纳米颗粒溶液冻干或喷雾干燥,得到水溶性He。
步骤(1)中,所述的SPI溶液通过包括以下步骤的方法制备:将SPI直接分散到pH11.0~12.0的氢氧化钠溶液中;或将SPI先分散在水中,再用氢氧化钠溶液调节pH至11.0~12.0。
步骤(2)中,所述的He为源自猪血的血红素或血红素衍生化合物。
步骤(2)中,He的加入量最高达到1.0%(w/v)。
步骤(2)中,优选使用盐酸溶液调节pH,如使用1.0mol/L的盐酸溶液。
步骤(3)中,SPI-He复合纳米颗粒溶液经透析处理除盐和离子后,再冻干或喷雾干燥。
一种可溶性He,通过上述方法得到。本发明的可溶性He,即SPI和He的纳米复合物易溶于水,乳化能力强,可用于生产He铁强化型豆奶或其它生物铁强化型液态饮品、固态饮品、代餐粉等各类食品。这种水溶性的He产品可以作为补铁剂广泛应用于各类食品或医药中。
本发明利用SPI在碱性条件下溶解,从而使He在相同体系下充分溶解,然后在缓慢调节pH至中性的过程中,SPI的疏水性结构逐渐暴露,与此同时He分子与SPI的疏水区域发生结合,从而同时掩蔽了SPI和He的疏水结构,而亲水结构暴露在水中,因此同时促进了SPI和He在中性水溶液中的溶解。本发明拓宽了He在食品中的应用形式和应用范围。
本发明有以下优点:
(1)产品中He易溶于水;
(2)产品中SPI也易溶于水(原料SPI为碱溶性);
(3)产品可用作补铁功能食品的优质原料;
(4)产品可用作生产液态食品的原料;
(5)产品中He的稳定性明显提高;
(6)产品中He含量高;
(7)产品具有良好的乳化性和乳化稳定性,可用于生产乳化类食品。
附图说明
图1为大豆分离蛋白(SPI)和氯化血红素(He)形成的复合物纳米颗粒形态和平均粒径。样品均为pH7.0的SPI-He复合物纳米颗粒溶液透析后样品,其中SPI初始浓度均为1.0%(w/v),氯化血红素初始浓度依次为0、0.1、0.25、0.5、1.0%(w/v),图1a、b、c、d、e为血红素初始浓度不同的各样品在液氮中先进行玻璃化处理,然后冻干,通过场发射扫描电子显微镜放大50000倍观察纳米颗粒形态。图f为上述透析过的SPI-He复合物溶液中纳米颗粒的平均粒径和多分散性指数。
图2为SPI-He复合纳米颗粒的溶解性。图2A各管样品中SPI初始浓度均为1.0%,pH值均为7.0,He初始浓度从左到右依次为:0、0.1、0.25、0.5、1.0%(w/v),均通过离心除去不溶性沉淀所得上清液,倒置观察,He含量越高,溶液颜色越深。图2B中样品与图2A中样品一致。图2C为图2A中的各上清液冻干后,以冻干粉初始浓度为0.3%(w/v)复溶于pH7.0的10mmol/L磷酸氢钠-磷酸二氢钠缓冲溶液中,然后10000rpm离心20min,取上清液进行紫外-可见光扫描,最大吸收波长393nm处的吸光值越大表明血红素含量越高,其中样品0.25%He为不含SPI的对照,操作与其它各样品相同。
图3为SPI-He复合物乳液室温贮藏过程中的稳定性。各乳液中pH7.0的SPI-He复合纳米颗粒溶液的体积分数为90%,油相为中链脂肪酸,体积分数为10%。制备SPI-He复合纳米颗粒溶液的SPI浓度均为1.0%(w/v),He浓度依次为0、0.01%、0.02%、0.03%(w/v)。
具体实施方式
本发明提供了一种溶解血红素的方法,具备包括以下步骤:
(1)将SPI加入至去离子水中,室温搅拌,使蛋白充分分散于水中,得到1.0~2.0%的蛋白分散液,用氢氧化钠溶液调节蛋白分散液pH至11.0~12.0,搅拌使蛋白充分溶解,得到SPI碱性水溶液。
或:将SPI分散于pH 11.0~12.0的氢氧化钠溶液中,搅拌使蛋白充分溶解,制备得到1.0~2.0%的SPI碱性水溶液。
(2)将源自猪血的氯化血红素固体颗粒慢慢加入至SPI碱性水溶液中,搅拌溶解,得到SPI、He混合碱性水溶液。
(3)立即用1.0mol/L盐酸溶液慢慢调节pH值至6.5~7.5,得到SPI-He复合纳米颗粒溶液。
(4)将SPI-He复合纳米颗粒溶液透析,除去盐和离子,最后冻干或喷雾干燥,制得水溶性He。
下面结合具体实施例进一步说明本发明的优点和效果。
实施例1
(1)将SPI加入至去离子水中,室温搅拌,使蛋白充分分散于水中,得到1.0%的SPI分散液。
(2)用1.0mol/L的氢氧化钠溶液调节SPI分散液的pH值至11.0~12.0,连续搅拌使蛋白充分溶解,得到SPI碱性水溶液。
(3)将SPI碱性水溶液分成5份,慢慢加入源自猪血的氯化血红素固体颗粒分别至浓度为0、0.1%、0.25%、0.5%、1.0%(w/v),边加边搅拌使血红素完全溶解,得到SPI、He混合碱性水溶液。
(4)立即用1.0mol/L盐酸溶液慢慢调节各溶液pH值至7.0,得到SPI-He复合纳米颗粒溶液。
(5)将各SPI-He复合纳米颗粒溶液透析,除去盐和离子,最后冻干。
(6)利用场发射扫描电子显微镜(Hitachi SU8010,日本东京)观察冻干粉复合颗粒的微观形貌,如图1a、b、c、d、e。如图1a所示,不含血红素的SPI有明显的颗粒聚集现象,而含有血红素的图1b、c、d、e形成了分散性较好的纳米颗粒,随着血红素浓度增加,纳米颗粒的粒度有逐渐增大的趋势。
(7)对透析后的SPI-He复合纳米颗粒溶液,利用纳米粒度仪Zetasizer Nano Zs+MPT-2(Malven Instruments Ltd.,Worcestershire,UK)测定溶液中颗粒的粒径,如图1f所示。可以看出,在含有He的SPI体系,SPI和He发生了结合,形成了纳米颗粒,并且随着He浓度增加,纳米颗粒的粒径有逐渐增大的趋势。
实施例2
(1)将SPI加入至去离子水中,室温搅拌,使蛋白充分分散于水中,得到1.0%的SPI分散液。
(2)用1.0mol/L的氢氧化钠溶液调节SPI分散液的pH值至11.0~12.0,连续搅拌使蛋白充分溶解,得到SPI碱性水溶液。
(3)将SPI碱性水溶液分成5份,慢慢加入源自猪血的氯化血红素固体颗粒分别至浓度为0、0.1%、0.25%、0.5%、1.0%(w/v),边加边搅拌使血红素完全溶解,得到SPI、He混合碱性水溶液。
(4)立即用1.0mol/L盐酸溶液慢慢调节各溶液pH值至7.0,得到SPI-He复合纳米颗粒溶液,10000rpm离心20min,取上清液,倒置观察,如图2A所示,颜色越深表明血红素含量越高。
(5)测定(4)中各上清液在393nm处的吸光值(A393),并根据标准曲线对溶液中的血红素浓度进行定量,通过下列公式计算出各样品中血红素的溶解百分比。其中C0为各溶液中初始血红素浓度,C为溶液中溶解的血红素浓度。结果如图2B所示,当添加的血红素初始浓度为0.5%或以下时,80%以上的血红素可以与SPI发生结合,从而溶解在复合体系中。当血红素初始浓度为1.0%(w/v)时,未与SPI结合的血红素分子明显增多,因此溶解百分比降低,约为60%。
(5)再将(4)中各SPI-He复合纳米颗粒上清液溶液透析,除去盐和离子,最后冻干。
(6)将(5)得到的冻干粉以初始浓度0.3%(w/v)复溶于pH7.0的10mmol/L磷酸氢钠-磷酸二氢钠缓冲溶液中,然后10000rpm离心20min,取上清液利用紫外-可见分光光度计(UV-2600,Shimadzu,Kyoto,Japan)扫描300~600nm范围内的吸收光谱,如图2C所示,300~450nm范围内的吸收峰越强表明血红素含量越高,其中样品0.25%He为不含SPI的对照,无吸收峰,表明血红素基本不溶于水。
(7)图2试验结果表明,当SPI初始浓度为1.0%,氯化血红素的溶解度可达0.6%(w/v),而同法操作下无SPI的氯化血红素复溶时几乎不溶于水。
实施例3
(1)将SPI加入至去离子水中,室温搅拌,使蛋白充分分散于水中,得到1.0%的SPI分散液。
(2)用1.0mol/L的氢氧化钠溶液调节SPI分散液的pH值至11.0~12.0,连续搅拌使蛋白充分溶解,离心取上清液,得到SPI碱性水溶液。
(3)将SPI碱性水溶液分成4份,慢慢加入源自猪血的氯化血红素固体颗粒分别至浓度为0、0.01、0.02、0.03%(w/v),边加边搅拌使血红素完全溶解,得到SPI、He混合碱性水溶液。
(4)立即用1.0mol/L盐酸溶液慢慢调节各溶液pH值至7.0,10000rpm离心20min取上清液,得到SPI-He复合纳米颗粒溶液。
(5)按照体积分数为10%(v/v)的比例在SPI-He复合纳米颗粒溶液中复合物溶液加入中链脂肪酸(MCT),加入叠氮化钠至浓度为0.04%(w/v),以抑制贮藏期间微生物的生长繁殖。用高速均质乳化机(FA25-D,Fluko,中国上海)13000rpm高速剪切3min,然后立即用高压均质机(AH-1500,Boyn Instrument Co.,Ltd.,中国杭州)经过3次循环制备SPI-Heme乳液。
(6)将SPI-Heme乳液于室温贮藏,每隔7天观察乳析情况并拍照,如图3(左)所示,同时取样用激光共聚焦显微镜(CLSM)显微观察油滴分布和聚集情况,如图3(右)所示。图3显示,同样条件下,SPI-He纳米复合物乳液的稳定性普遍高于SPI乳液。
应理解,本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (8)
1.一种溶解血红素的方法,其特征在于:包括以下步骤:
(1)制备大豆分离蛋白浓度为1.0~2.0%、pH为11.0~12.0的大豆分离蛋白溶液;
(2)将血红素或血红素衍生化合物加入到大豆分离蛋白溶液中,完全溶解后,调节pH至6.5~7.5,得到大豆分离蛋白-血红素复合纳米颗粒溶液;
(3)将大豆分离蛋白-血红素复合纳米颗粒溶液冻干或喷雾干燥,得到可溶性血红素。
2.根据权利要求1所述的溶解血红素的方法,其特征在于:步骤(1)中,所述的大豆分离蛋白溶液通过包括以下步骤的方法制备:将大豆分离蛋白直接分散到pH 11.0~12.0的氢氧化钠溶液中,充分溶解;或将大豆分离蛋白先分散在水中,再用氢氧化钠溶液调节pH至11.0~12.0。
3.根据权利要求1所述的溶解血红素的方法,其特征在于:步骤(2)中,所述的血红素或血红素衍生化合物为源自猪血的血红素或血红素衍生化合物。
4.根据权利要求1所述的溶解血红素的方法,其特征在于:步骤(2)中,血红素的加入量为0.1%~1.0%。
5.根据权利要求1所述的溶解血红素的方法,其特征在于:步骤(2)中,使用盐酸溶液调节pH。
6.根据权利要求1所述的溶解血红素的方法,其特征在于:步骤(3)中,大豆分离蛋白-血红素复合纳米颗粒溶液经透析处理除盐和离子后,再冻干或喷雾干燥。
7.一种可溶性血红素,其特征在于:通过权利要求1-6任一项所述的方法得到。
8.权利要求7所述的可溶性血红素在食品或医药中的应用。
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