CN113892652A - 一种油溶性益生菌微胶囊复合冻干粉及其制备方法 - Google Patents
一种油溶性益生菌微胶囊复合冻干粉及其制备方法 Download PDFInfo
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- CN113892652A CN113892652A CN202111190820.8A CN202111190820A CN113892652A CN 113892652 A CN113892652 A CN 113892652A CN 202111190820 A CN202111190820 A CN 202111190820A CN 113892652 A CN113892652 A CN 113892652A
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Abstract
本发明提供一种油溶性益生菌微胶囊复合冻干粉及其制备方法,本发明通过微胶囊技术将活性益生菌包裹在具有囊壁结构的微胶囊中,一方面,本发明的微胶囊囊壁结构具有很好的机械性能,能够很好地将活性益生菌成功包裹到囊壁结构中,克服了活菌易死亡,菌数不稳定,导致益生菌产品保质期短,质量不稳定的技术问题,另一方面,本发明的油溶性益生菌进入人体胃部后,囊壁结构能够保证在胃液中不分解,而在肠液中能快速分解释放益生菌,使活性益生菌能够快速有效被吸收而发生作用。
Description
技术领域
本发明涉及营养制品技术领域,尤其涉及一种油溶性益生菌微胶囊复合冻干粉及其制备方法。
背景技术
随着人们生活水平的提高,对健康的要求越来越高,益生菌普遍的被接受并广泛使用。益生菌是一类对宿主有益的活性微生物,是定植于人体肠道、生殖系统内,能产生确切健康功效从而改善宿主微生态平衡、发挥有益作用的活性有益微生物的总称。益生菌的概念在1989年被提出,2002年科学家确定益生菌的概念和疗效。之后短短十年世界人类卫生领域发生了巨大的变化,益生菌益生元的研究和产品开发成为了潮流。该领域的研发、生产,目前在乳酸菌、双歧杆菌的研究和应用方面尤其突出。这方面的产品有传统的酸奶,以及新开发的各种酸奶,乳酸饮料,各种单复合菌保健品、食品,还有用于治疗肠道疾病的益生菌制剂。因为益生菌的保存条件苛刻,现有产品普遍存在着活菌易死亡,菌数不稳定,导致益生菌产品保质期短,质量不稳定的技术问题。
发明内容
本发明的目的是为了解决上述现有技术的不足,而提出一种油溶性益生菌微胶囊复合冻干粉及其制备方法。
为了实现上述目的,本发明提供一种油溶性益生菌微胶囊复合冻干粉,其特征在于:包含油溶性益生菌微胶囊、乳清浓缩蛋白粉、低聚异麦芽糖、菊粉和低聚木糖,所述油溶性益生菌微胶囊包含壁材和囊芯物质,所述囊芯物质为油溶性益生菌,所述壁材为乙酸丙酯京尼平与明胶海藻酸钠交联后形成的聚合物,所述聚合物的化学结构式如式(Ⅰ)所示:
式(Ⅰ)中:R1至R6选自十八种不同氨基酸甘氨酸、丙氨酸、丝氨酸、天门冬氨酸、谷氨酸氨基、脯氨酸、精氨酸、组氨酸、酪氨酸、胱氨酸、亮氨酸、苏氨酸、蛋氨酸、缬氨酸、苯丙氨酸、色氨酸、谷氨酸、赖氨酸的残基中的一种。
优选地,所述式(Ⅰ)中:
R6选自十八种不同氨基酸甘氨酸、丙氨酸、丝氨酸、天门冬氨酸、谷氨酸氨基、脯氨酸、精氨酸、组氨酸、酪氨酸、胱氨酸、亮氨酸、苏氨酸、蛋氨酸、缬氨酸、苯丙氨酸、色氨酸、谷氨酸、赖氨酸的残基中的一种;
R1、R5为丝氨酸、苏氨酸或酪氨酸的残基;
R2、R4为天门冬氨酸或谷氨酸氨基的残基;
R3为赖氨酸或者精氨酸的残基。
优选地,所述油溶性益生菌包含有:不饱和脂肪酸、糖类、多元醇、脂肪醇、碱性氨基酸、共轭多稀酸、益生菌。
优选地,所述油溶性益生菌包含有:核桃油或调和油、益生菌菌粉、益生元、防粘抗氧化剂。
优选地,所述油溶性益生菌包含有长双歧杆菌、短双歧杆菌、青春双歧杆菌、嗜热链球菌、保加利亚乳杆菌、乳酸乳杆菌中的一种或几种。
本发明还提供一种油溶性益生菌微胶囊复合冻干粉的制备方法,具体包括如下步骤:
a.将海藻酸钠溶解于水中,得到海藻酸钠水溶液;
b.将明胶溶于水中,得到明胶水溶液;
c.将脂肪酸蔗糖酯和大豆磷脂溶于油溶性益生菌溶液中,得到油性混合溶液;
d.将步骤c的油性混合溶液加入到步骤b的明胶水溶液中,充分乳化;
e.在步骤d乳化后的体系中加入海藻酸钠溶液,水浴加热并搅拌均匀;
f.关闭加热冷却至室温,向步骤e的反应体系加入氯化钙溶液,在25-30℃下反应完全;
g.调节体系pH值至8-9,向体系中加入京尼平水溶液,室温下反应完全;
h.在碱性条件下,加热并向反应体系中加入乙酸丙酯,反应完全后得到油溶性益生菌微胶囊;
i.将油溶性益生菌微胶囊与乳清浓缩蛋白粉、低聚异麦芽糖、菊粉、低聚木糖混合,经过真空冷冻干燥,得到益生菌微胶囊复合冻干粉。
优选地,其特征在于,所述步骤d中,乳化温度为37℃,乳化时间为60min,搅拌速率为600rpm。
优选地,其特征在于:所述步骤e和步骤f之间还包括:控制步骤e的水浴温度为37℃,调节步骤e的反应体系的pH值至3-4,搅拌速率调节为800-900rpm,继续搅拌60min;关闭加热,使体系温度降至室温,再使用氨水溶液调节体系pH至6-7,搅拌均匀。
优选地,其特征在于:所述步骤f中,加入的氯化钙溶液浓度为0.015-0.040mol/L,反应时间为60min;所述步骤g中还包括加入质量分数为0.5%的京尼平水溶液后先25-30℃反应3小时,然后升温至37℃反应10-12小时;所述步骤h中向反应体系中加入乙酸丙酯后,还包括在反应开始后向反应体系中加入醋酸作为催化剂,温度为40℃,反应完全后得到油溶性益生菌微胶囊;所述步骤i中的真空冷冻干燥预具体条件为:预冷冻温度为-40℃,时间2h;干燥升华阶段,干燥温度为15℃,真空度为0.09-2MPa,干燥时间为25-30h。
优选地,所述步骤i中,按照重量份数计,油溶性益生菌微胶囊50-60份、乳清浓缩蛋白粉15-25份、低聚异麦芽糖1-6份、菊粉1-6份、低聚木糖1-6份。
优选地,按照重量份数计,各组分的配比为:2.0-3.5%明胶溶液60-70份,2.0-3.5%海藻酸钠溶液60-70份,油性益生菌混合物20-30份,脂肪酸蔗糖酯0.2-0.4份,大豆磷脂0.15-0.25份,氯化钙6-10份,0.5%京尼平8-12份,乙酸丙酯为2-5份。
进一步优选地,按照重量份数计,3.0%明胶溶液选择66份,3.0%海藻酸钠溶液选择66份,油性益生菌混合物选择25份,脂肪酸蔗糖酯0.3份,大豆磷脂选择0.2份,氯化钙选择8份,京尼平选择10份,乙酸丙酯选择5份。
与现有技术相比,本发明的有益效果是:本发明通过微胶囊技术将活性益生菌包裹在具有囊壁结构的微胶囊中,一方面,本发明的微胶囊囊壁结构具有很好的机械性能,能够很好地将活性益生菌成功包裹到囊壁结构中,克服了活菌易死亡,菌数不稳定,导致益生菌产品保质期短,质量不稳定的技术问题,另一方面,本发明的油溶性益生菌进入人体胃部后,囊壁结构能够保证在胃液中不分解,而在肠液中能快速分解释放益生菌,使活性益生菌能够快速有效被吸收而发生作用。另外,一方面,乳清浓缩蛋白粉作为益生菌微胶囊复合载体,其营养价值较高,可提供身体构造新组织所需的氨基酸,可改善肠胃功能。另一方面,益生菌冻干粉中包含的低聚异麦芽糖、菊粉和低聚木糖属于益生元,其不被宿主消化吸收,能够选择性地促进肠道益生菌的代谢和增殖,从而改善人体健康。
附图说明
图1为实施例1微胶囊TG和DTG曲线图;
图2为实施例1制备的油溶性益生菌在模拟胃液中透过率与时间的关系图。
图3为实施例1制备的微油溶性益生菌在模拟肠液中透过率与时间的关系图。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地说明。
实施例1油溶性益生菌的制备
本实施例对油溶性益生菌的详细制备方法说明如下:
a.称取2.0g的海藻酸钠于烧杯,加入66mL去离子水,室温下搅拌溶解;
b.称取2.0g的明胶于三口烧瓶,加入66mL去离子水,于45℃的水浴温度下溶解,得到3.0%明胶溶液;
c.取25mL的活性益生菌溶液,加入脂肪酸蔗糖酯和大豆磷脂,得到油相混合溶液;
d.将上述油性混合溶液加入到明胶溶液中进行乳化,乳化温度37℃,搅拌速度600rpm,乳化时间60min;
e.向步骤d的体系中加入海藻酸钠溶液,随着体系中海藻酸钠溶液增多,粘度增大,提高搅拌速度至800rpm;加完海藻酸钠溶液后,37℃水浴温度下搅拌60min;
此时,优选采用醋酸调节体系PH至3-4,搅拌30min;在体系pH调节为3-4的介质中,明胶的等电点为5,由于低于明胶的等电点,-NH3 +的含量大于-COO-,因此明胶分子量上带有大量的正电荷,海藻酸钠分子链上则带有大量负电荷的羧基,带正电的明胶与带负电的海藻酸钠发生静电吸引,微胶囊囊壁上形成聚电解质膜,使微胶囊稳定性提高;
明胶与海藻酸钠的反应式如下所示:
f.关闭加热,使体系温度降至室温,体系粘度增大,约60min;温度降至室温,使明胶因温度降低为发生凝胶反应,从而使微胶囊囊壁强度增大,稳定性提高;
此时再使用5%氨水溶液调节体系PH至6-7,继续搅拌30min;向体系缓慢滴加8mL,0.02mol/L CaCl2溶液,25-30℃反应60min;氯化钙与海藻酸钠上的Na+发生置换反应,钙离子通过与海藻酸钠上的羟基、氨基以及糖苷键产生配位作用,形成具有蛋壳结构的海藻酸钙,海藻酸钠与氯化钙反应式如下所示:
使用5%氨水溶液调节体系pH至8-9,搅拌30min;使明胶上的部分质子化氨基转为游离氨基,便于下一步与交联剂反应;
g.向体系中缓慢滴加10mL,0.5%京尼平溶液,在室温下反应3h,然后升温至37℃反应10-12小时,得到包油型微胶囊;其交联机理为水溶液中的-OH–亲核攻击京尼平,使含氧六元环生成一个醛基和一个烯醇结构,由于烯醇结构不稳定,转化为醛基,最终京尼平形成含双醛结构化合物,开环后的京尼平分子上的醛基可以与另一分子京尼平五元环上的羟基反应,发生分子间聚合,聚合后的京尼平与明胶上的氨基进行Schiff碱反应,形成长链京尼平交联桥的网状结构聚合物;具体反应式如下所示:
h.在碱性条件下,温度40℃,向反应体系中加入适量乙酸丙酯,反应20min,使乙酸丙酯水解,再向反应体系中加入醋酸,调节pH为2-3,水解生成的丙醇则与明胶中游离羧基反应,从而对囊壁进行改二次交联,提高囊壁强度,提高稳定性,得到油溶性益生菌;乙酸丙酯改性的微胶囊囊壁材料结构式如式(Ⅰ)所示:
i.将上述油溶性益生菌转移至广口瓶中,密封保存。
j.将油溶性益生菌微胶囊与乳清浓缩蛋白粉、低聚异麦芽糖、菊粉、低聚木糖混合,经过真空冷冻干燥,得到益生菌微胶囊复合冻干粉。
作为具体实施方式,本实施例中所述活性益生菌的配方如下:
配方1
不饱和脂肪酸(选自油酸/亚油酸/亚麻酸/花生四烯酸,其中的一种或多种)、糖类(选自蔗糖/海藻糖/乳糖/葡萄糖/麦芽糊糖/可溶性淀粉,其中的一种或多种)、多元醇(选自乙二醇/甘油/甘露醇/木糖醇/山梨醇/叔丁醇,其中的一种或多种)、脂肪醇(选自辛醇/癸醇/月桂醇/油醇,其中的一种或多种)、碱性氨基酸(选自L-精氨酸/赖氨酸/组氨酸,其中的一种或多种)、共轭多稀酸(选自桐油酸/共轭亚麻酸/共轭十八碳四烯酸,其中的一种或多种)、益生菌(选自链球菌属益生菌/乳杆菌属益生菌/双歧杆菌属益生菌,其中的一种或多种)。
配方2
核桃油或调和油(选自亚麻籽油/花生油/火麻油/菜籽油/大豆油/棉籽油/芝麻油/葵花籽油/棕榈油/油菜籽油/玉米油/橄榄油/茶油/芥花籽油/小麻油/胡麻油/牡丹籽油/棕榈油/椰子油/蘑菇油/DHA藻油,其中的一种或几种)、益生菌菌粉(选自嗜酸乳杆菌/动物双歧杆菌/乳双歧杆菌/鼠李糖糖乳杆菌/罗伊氏乳杆菌/发酵乳杆菌/短双岐杆菌,其中的一种或几种)、益生元(选自异麦芽低聚糖/低聚果糖/低聚半乳糖/低聚木糖/大豆低聚糖/菊粉植物中草药提取物/蛋白质水解物/多元醇,其中的一种或几种)、防粘抗氧化剂(选自中链甘油三酯/饱和辛酸甘油三酯/饱和癸酸甘油三酯/饱和辛酸甘油三酯与饱/癸酸甘油三酯复配物/L-抗坏血酸棕榈酸酯/维生素E/二氧化硅,其中的一种或几种)。
实施例2热重分析(TG)
明胶、海藻酸钠、实施例1制备的油溶性益生菌各称取6mg样品,采用TG热分析仪进行分析,以升温速率为10℃将温度从室温升温至600℃,气体氛围为氮气。得到明胶、海藻酸钠和实施例1油溶性益生菌的热分解温度表如表1所示,得到实施例1微胶囊TG和DTG曲线图如图1所示。
表1明胶、海藻酸钠和实施例1微胶囊的热分解温度
由表1可知经过的实施例1油溶性益生菌微胶囊的热稳定性低于明胶,但高于海藻酸钠,其原因为氯化钙与海藻酸钠反应生成海藻酸钙,而海藻酸钙在较低温度会分解形成CaCO3,CaCO3会进一步氧化形成CaO和Ca(OH)2,因此在比明胶低的温度开始分解。
图1中曲线1为TG曲线,曲线2为DTG曲线)由图1可见,实施例1油溶性益生菌的热分解存在三个阶段,第一阶段出现在75-100℃左右,在该阶段中,随着温度逐渐升高,实施例1油溶性益生菌中残存的水分逐渐蒸发,造成该阶段的失重。第二阶段出现在240-260℃附近,在这个阶段内随着温度的升高实施例1油溶性益生菌发生显著的失重,是油溶性益生菌中微胶囊囊壁材料热降解过程的主要过程,由DTG曲线发现在该阶段内出现分峰,其原因为海藻酸盐裂解为较稳定的中间产物,并伴随着部分糖苷键的断裂,相邻羟基以水分子的形式脱去以及明胶上肽键的断裂。第三个失重阶段发生在300-360℃之间,该阶段为中间产物继续分解,脱羧放出二氧化碳并部分炭化。
实施例3耐模拟胃酸试验
模拟胃液的配制:盐酸16.4mL,胃蛋白酶10g,加去离子水搅匀后定容至1000mL,pH=1.2。
以实施例1制备的油溶性益生菌在胃液中的溶解情况。称取0.05g实施例1制备的油溶性益生菌于烧杯中,加入30mL模拟胃液,于37℃的水浴条件的条件下进行模拟实验,每30min取样一次,每次取1.2mL溶液,加入2.5mL去离子水得到待测溶液。由于实施例1制备的油溶性益生菌在波长为600nm处存在京尼平与明胶上的氨基反应形成的交联键的特征吸收峰,通过测定其在波长600nm处的透过率,可分析交联膜在模拟胃液中的溶解情况。得到实施例1制备的油溶性益生菌在模拟胃液中透过率与时间的关系图如图2所示。
由图2可见,在模拟胃液中,溶液在波长600nm处的透过率随时间延长下降,并在时间为280min后,透过率变化基本平稳,其透过率为93%左右。油溶性益生菌经过模拟胃液处理390min后,透过率变化5%左右,说明油溶性益生菌在模拟胃液中的稳定性较好,具有较好的抵抗胃酸侵蚀的能力,可较好保护囊芯物质,使囊芯物质达到肠液中仍有较高的活性。
实施例4耐模拟肠液试验
模拟肠液的制备:称取磷酸二氢钾6.8g,加去离子水500mL溶解,用0.4%的氢氧化钠溶液调节pH值至6.8,另取胰蛋白酶10g加去离子水适量使其溶解,将两液混合后,加去离子水定容至1000mL。
以实施例1制备的油溶性益生菌在模拟肠液中的溶解情况。称取0.05g实施例1制备的油溶性益生菌于烧杯中,加入30mL模拟肠液,于37℃水浴条件下进行模拟实验,每3min取样一次,每次取1.2mL溶液,加入2.5mL去离子水得到待测溶液。通过测定其在波长600nm处的透过率,可分析油溶性益生菌在模拟肠液的溶解情况。得到实施例1制备的油溶性益生菌在模拟肠液中透过率与时间的关系图如图3所示。
由下图3可见,当实施例1制备的油溶性益生菌加入模拟肠液中后,溶液的透过率随着时间的延长呈快速下降趋势,说明油溶性益生菌经模拟肠液处理迅速溶解,当时间达到9min后,透过率变化趋于平稳,透过率在83%左右,并且烧杯中观察到油溶性益生菌已溶解完全,形成均一深蓝色溶液,说明油溶性益生菌溶解完全。由此说明,油溶性益生菌的微胶囊在肠液中会发生溶解破裂,稳定性较差,有利于囊芯物质的释放。
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揭露的技术范围内,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 (11)
2.根据权利要求1所述油溶性益生菌微胶囊复合冻干粉,其特征在于:所述式(Ⅰ)中:
R6选自十八种不同氨基酸甘氨酸、丙氨酸、丝氨酸、天门冬氨酸、谷氨酸氨基、脯氨酸、精氨酸、组氨酸、酪氨酸、胱氨酸、亮氨酸、苏氨酸、蛋氨酸、缬氨酸、苯丙氨酸、色氨酸、谷氨酸、赖氨酸的残基中的一种;
R1、R5为丝氨酸、苏氨酸或酪氨酸的残基;
R2、R4为天门冬氨酸或谷氨酸氨基的残基;
R3为赖氨酸或者精氨酸的残基。
3.根据权利要求1所述油溶性益生菌微胶囊复合冻干粉,其特征在于:所述油溶性益生菌包含有:不饱和脂肪酸、糖类、多元醇、脂肪醇、碱性氨基酸、共轭多稀酸、益生菌。
4.根据权利要求1所述油溶性益生菌微胶囊复合冻干粉,其特征在于:所述油溶性益生菌包含有:核桃油或调和油、益生菌菌粉、益生元、防粘抗氧化剂。
5.根据权利要求1所述油溶性益生菌微胶囊复合冻干粉,其特征在于:所述油溶性益生菌包含有长双歧杆菌、短双歧杆菌、青春双歧杆菌、嗜热链球菌、保加利亚乳杆菌、乳酸乳杆菌中的一种或几种。
6.一种油溶性益生菌微胶囊复合冻干粉的制备方法,具体包括如下步骤:
a.将海藻酸钠溶解于水中,得到海藻酸钠水溶液;
b.将明胶溶于水中,得到明胶水溶液;
c.将脂肪酸蔗糖酯和大豆磷脂溶于油溶性益生菌溶液中,得到油性混合溶液;
d.将步骤c的油性混合溶液加入到步骤b的明胶水溶液中,充分乳化;
e.在步骤d乳化后的体系中加入海藻酸钠溶液,水浴加热并搅拌均匀;
f.关闭加热冷却至室温,向步骤e的反应体系加入氯化钙溶液,在25-30℃下反应完全;
g.调节体系pH值至8-9,向体系中加入京尼平水溶液,室温下反应完全;
h.在碱性条件下,加热并向反应体系中加入乙酸丙酯,反应完全后得到油溶性益生菌微胶囊;
i.将油溶性益生菌微胶囊与乳清浓缩蛋白粉、低聚异麦芽糖、菊粉、低聚木糖混合,经过真空冷冻干燥,得到益生菌微胶囊复合冻干粉。
7.根据权利要求6所述一种油溶性益生菌微胶囊复合冻干粉的制备方法,其特征在于,所述步骤d中,乳化温度为37℃,乳化时间为60min,搅拌速率为600rpm。
8.根据权利要求6所述的一种油溶性益生菌微胶囊复合冻干粉的制备方法,其特征在于:所述步骤e和步骤f之间还包括:控制步骤e的水浴温度为37℃,调节步骤e的反应体系的pH值至3-4,搅拌速率调节为800-900rpm,继续搅拌60min;关闭加热,使体系温度降至室温,再使用氨水溶液调节体系pH至6-7,搅拌均匀。
9.根据权利要求6所述的一种油溶性益生菌微胶囊复合冻干粉的制备方法,其特征在于:所述步骤f中,加入的氯化钙溶液浓度为0.015-0.040mol/L,反应时间为60min;所述步骤g中还包括加入质量分数为0.5%的京尼平水溶液后先25-30℃反应3小时,然后升温至37℃反应10-12小时;所述步骤h中向反应体系中加入乙酸丙酯后,还包括在反应开始后向反应体系中加入醋酸作为催化剂,温度为40℃,反应完全后得到油溶性益生菌微胶囊;所述步骤i中的真空冷冻干燥预具体条件为:预冷冻温度为-40℃,时间2h;干燥升华阶段,干燥温度为15℃,真空度为0.09-2MPa,干燥时间为25-30h。
10.根据权利要求6所述的一种油溶性益生菌微胶囊复合冻干粉的制备方法,其特征在于,所述步骤i中,按照重量份数计,油溶性益生菌微胶囊50-60份、乳清浓缩蛋白粉15-25份、低聚异麦芽糖1-6份、菊粉1-6份、低聚木糖1-6份。
11.根据权利要求6所述乙酸丙酯改性的包油型微胶囊的制备方法,其特征在于,按照重量份数计,各组分的配比为:2.0-3.5%明胶溶液60-70份,2.0-3.5%海藻酸钠溶液60-70份,油性益生菌混合物20-30份,脂肪酸蔗糖酯0.2-0.4份,大豆磷脂0.15-0.25份,氯化钙6-10份,0.5%京尼平8-12份,乙酸丙酯为2-5份。
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