CN110584104B - 一种改善大学生亚健康状态的专用营养补充剂制备方法 - Google Patents
一种改善大学生亚健康状态的专用营养补充剂制备方法 Download PDFInfo
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Abstract
本发明属于轻工食品加工的技术领域,具体的涉及一种改善大学生亚健康状态的专用营养补充剂制备方法。该种改善大学生亚健康状态的专用营养补充剂制备方法,包括以下步骤:(1)原料的破壁处理;(2)水溶功能性多糖溶液的制备;(3)水溶功能性多糖溶液的发酵处理;(4)复合凝聚体系的制备;(5)微胶囊湿囊的制备;(6)专用营养补充剂的制备。通过该制备方法制得的营养补充剂在改善易疲劳症状、保证营养方面具有一定的功效,有助于大学生强健体魄、活跃思维、提高记忆力,使其更有效率地学习和工作。
Description
技术领域
本发明属于轻工食品加工的技术领域,具体的涉及一种改善大学生亚健康状态的专用营养补充剂制备方法。
背景技术
按照世界卫生组织(WHO)界定,人体状态分为健康、疾病以及介于二者之间的低质状态,即“亚健康”。“亚健康”状态是心理、生理、社会三方面因素导致的机体神经系统、内分泌系统、免疫系统等整体协调失衡、功能紊乱,虽未引起器质性的疾病,但机体功能上表现出免疫力降低,抗病能力下降,失眠健忘,易患流行病等症状,甚至出现猝死现象。据WHO全球性调查结果显示,目前世界上75%的人处于“亚健康”状态,真正的健康群体不足5%,“亚健康”的全球性、普遍性已对人类的健康及生活质量构成了严重的威胁,成为当今全球性的重大社会问题。
众多数据表明,当前大学生出现“亚健康”状态与因日常饮食不规律所导致的营养缺乏、不均衡等问题密切相关。营养素是保证人类健康的物质基础,营养素的均衡搭配是人体健康的重要保障,种种迹象表明合理饮食在降低疾病风险、辅助治疗特殊群体方面功效显著。因此合理的营养补充有益于人体所需营养素间的均衡搭配,是改善“亚健康”状态最行之有效的办法之一。
发明内容
本发明的目的在于针对目前大学生群体所普遍存在的“亚健康”症状而提供一种改善大学生亚健康状态的专用营养补充剂制备方法,通过该制备方法制得的营养补充剂在改善易疲劳症状、保证营养方面具有一定的功效,有助于大学生强健体魄、活跃思维、提高记忆力,使其更有效率地学习和工作。
本发明的技术方案为:一种改善大学生亚健康状态的专用营养补充剂制备方法,包括以下步骤:
(1)原料的破壁处理:将作为原料的中药材进行破壁处理,制得中药粉体;
(2)水溶功能性多糖溶液的制备:首先将步骤(1)所得中药粉体溶解后加入复合酶进行酶解,得到酶解液;然后将酶解液离心取上清液,即得水溶功能性多糖溶液;
(3)水溶功能性多糖溶液的发酵处理:在步骤(2)所得的水溶功能性多糖溶液中加入活化后的复合乳酸菌进行发酵处理,得到发酵液;
(4)复合凝聚体系的制备:将步骤(3)所得的发酵液与脂溶性活性成分和乳化剂匀浆制得复合凝聚体系;
(5)微胶囊湿囊的制备:首先将超微处理后的蛋白与步骤(4)所得复合凝聚体系进行均质处理;然后静置,制备得到均一的微胶囊分散液,离心除去上清液得到微胶囊湿囊;
(6)专用营养补充剂的制备:对步骤(5)所得的微胶囊湿囊进行冷冻干燥,制得专用营养补充剂。
所述步骤(1)中破壁处理采用气流超微粉碎,其中气流超微粉碎的气流压力为2~3MPa,转速为2500~4000r/min;粉碎后过400~600目筛。
所述步骤(1)中的中药材包括枸杞、淫羊藿、红景天和丹参;按照质量比枸杞:淫羊藿:红景天:丹参为35~45:4~10:20~30:25~30。
所述步骤(2)将中药粉体溶解于水中,中药粉体与水的质量比为3~5:5~7;所述复合酶的添加量为中药粉体加入量的2~8%;所述酶解的酶解时间为8~10h,酶解温度为35~50℃,pH值控制在3.5~5;所述离心的离心转速为2000~4000r/min,离心时间为30~50min。
所述步骤(2)中复合酶由果胶酶、纤维素酶和淀粉酶按照质量比3~5:6~9:2~4复合而成。
所述步骤(3)中复合乳酸菌的添加量为水溶功能性多糖溶液总质量的2%~5%;发酵温度为30~39℃,发酵时间为8h~12h。
所述步骤(3)中复合乳酸菌为嗜酸乳杆菌、保加利亚乳杆菌、婴儿双歧杆菌、德氏乳杆菌和鼠李糖乳杆菌中的至少一种。
所述步骤(4)中匀浆采用高速匀浆机,匀浆转速为8000~15000r/min,匀浆时间为20~30min;所述脂溶性活性成分由维生素A、维生素D3、维生素E、维生素K2、人体必需矿物质、高活性氨基酸和ω-3-不饱和脂肪酸有机复配而成,且维生素A的添加量为发酵液质量的3%~5%,维生素D3的添加量为发酵液质量的1%~3%,维生素E的添加量为发酵液质量的5%~8%,维生素K2的添加量为发酵液质量的2%~4%,人体必需矿物质的添加量为发酵液质量的6%~10%,高活性氨基酸的添加量为发酵液质量的8%~15%,ω-3-不饱和脂肪酸的添加量为发酵液质量的6%~12%;其中人体必需矿物质由葡萄糖酸镁、葡萄糖酸锌和葡萄糖酸铁按质量比1~2:1~4:1~3组成;高活性氨基酸由精氨酸、谷氨酸、甘氨酸和γ-氨基丁酸按质量比1~3:3~5:2~4:1~3组成;ω-3-不饱和脂肪酸由DHA和EPA按质量比2~4:3~7组成;所述乳化剂为聚氧乙烯氧丙烯油酸酯、聚氧乙烯单月桂酸酯、失水山梨醇月桂酸酯、二乙二醇脂肪酸酯、聚氧乙烯蓖麻油和聚氧乙烯植物油中的至少一种,其中乳化剂的添加量为发酵液与脂溶性活性成分混合总量的0.5%~3.5%,HLB值控制在9~12。
所述步骤(5)中超微处理选用气流式粉碎机,粉碎压力为0.7~1.5MPa,转速为1500~2000r/min,粉碎后过200~600目筛分离、细化;所述蛋白与复合凝聚体系的质量比为3~6:4~7;所述均质处理采用高压均质,均质压力为20~45MPa,均质时间为15~20min;所述静置的时间为8~12h;所述蛋白为大豆分离蛋白、乳清分离蛋白、β-乳球蛋白及β-伴大豆球蛋白中的至少一种。
所述步骤(6)中预冻温度为-50~-30℃,物料厚度为5~15mm,冷冻干燥时间为20~35h。
本发明的有益效果为:本发明针对大学生“亚健康”症状,巧妙运用中医“寓医于食”相关理论,通过药性配伍、阴阳相济的原则优选富含大学生“亚健康”所缺营养素的“药食同源”类物质为基础原料,结合物理破壁;生物酶解,利用绿色、高效复合酶解工艺获取高活性多糖;乳酸菌生物转化,引入具有改善肠道菌群结构的乳酸菌,提高营养剂的消化吸收率;复合凝聚体系构建、基于蛋白与还原性糖间的美拉德反应制备微胶囊及冷冻干燥技术获取高活性、高稳定型营养补充剂;最终得到全营养、高功效的大学生亚健康专用营养补充剂。该营养补充剂可有效实现活性成分的快速释放、大学生肠道菌群结构的改善及人体易吸收活性小分子的高效转化等,进而达到大学生“亚健康”群体所需营养素及时补充的目的,而且产品方便易携带,特色鲜明、针对性强,对维持我国大学生群体的营养均衡、提高免疫力具有显著功效,意义重大。
附图说明
图1为丹酚酸B标准样品液相色谱图。
图2为丹酚酸B标准曲线图。
图3为实施例1、对比例1和对比例2分别制备所得的营养补充剂释放丹酚酸B的液相色谱对比图。
图4为实施例1-3制备所得的营养补充剂与现有市售产品关于肠液中ω-3-不饱和脂肪酸累积释放率的对比图。
具体实施方式
下面通过实施例对本发明进行详细描述。
实施例1
所述改善大学生亚健康状态的专用营养补充剂制备方法,具体步骤如下:
(1)原料的破壁处理:对作为原料的中药材在气流压力为2MPa,转速为2500r/min条件下进行气流超微粉碎,粉碎后过400目筛制得中药粉体;其中中药材包括枸杞、淫羊藿、红景天和丹参;按照质量比枸杞:淫羊藿:红景天:丹参为45:5:20:30;
(2)水溶功能性多糖溶液的制备:首先取200g步骤(1)所得中药粉体溶解于水中,中药粉体与水的质量比为4:6;溶解后加入由果胶酶、纤维素酶和淀粉酶按照质量比3:8:2复合而成的复合酶进行酶解,酶解时间为8h,酶解温度为35℃,pH值控制在3.5~5,复合酶的添加量为中药粉体加入量的4%,得到酶解液;然后将酶解液在离心转速为2000r/min条件下离心30min,取上清液,即得水溶功能性多糖溶液;
(3)水溶功能性多糖溶液的发酵处理:在步骤(2)所得的水溶功能性多糖溶液中加入活化后的复合乳酸菌进行发酵处理,发酵温度为35℃,发酵时间为8h,得到发酵液;复合乳酸菌的添加量为水溶功能性多糖溶液总质量的3%;复合乳酸菌为保加利亚乳杆菌和婴儿双歧杆菌;
(4)复合凝聚体系的制备:将步骤(3)所得的发酵液与脂溶性活性成分和乳化剂加入至高速匀浆机,在匀浆转速为8000r/min条件下匀浆20min制得复合凝聚体系;所述脂溶性活性成分由维生素A、维生素D3、维生素E、维生素K2、人体必需矿物质、高活性氨基酸和ω-3-不饱和脂肪酸有机复配而成,且维生素A的添加量为发酵液质量的3%,维生素D3的添加量为发酵液质量的2%,维生素E的添加量为发酵液质量的6%,维生素K2的添加量为发酵液质量的4%,人体必需矿物质的添加量为发酵液质量的7%,高活性氨基酸的添加量为发酵液质量的10%,ω-3-不饱和脂肪酸的添加量为发酵液质量的6%;其中人体必需矿物质由葡萄糖酸镁、葡萄糖酸锌和葡萄糖酸铁按质量比1:1:1组成;高活性氨基酸由精氨酸、谷氨酸、甘氨酸和γ-氨基丁酸按质量比1:3:2:1组成;ω-3-不饱和脂肪酸由DHA和EPA按质量比2:3组成;所述乳化剂由二乙二醇脂肪酸酯和聚氧乙烯植物油按质量比1:2混合而成,其中乳化剂的添加量为发酵液与脂溶性活性成分混合总量的1%,HLB值控制在9;
(5)微胶囊湿囊的制备:首先选用气流式粉碎机在粉碎压力为0.8MPa,转速为1500r/min条件下将蛋白进行超微粉碎处理,粉碎后过200目筛分离、细化,再与步骤(4)所得复合凝聚体系进行高压均质处理,均质压力为20MPa,均质时间为15min;然后室温下静置8h,制备得到均一的微胶囊分散液,离心除去上清液得到微胶囊湿囊;所述蛋白与复合凝聚体系的质量比为3:7;所述蛋白由乳清分离蛋白、β-乳球蛋白及β-伴大豆球蛋白按质量比1.5:1:1混合而成;
(6)专用营养补充剂的制备:对步骤(5)所得的微胶囊湿囊进行冷冻干燥,预冻温度为-37℃,物料厚度为15mm,冷冻干燥时间为20h,制得专用营养补充剂。
实施例2
所述改善大学生亚健康状态的专用营养补充剂制备方法,具体步骤如下:
(1)原料的破壁处理:对作为原料的中药材在气流压力为3MPa,转速为4000r/min条件下进行气流超微粉碎,粉碎后过600目筛制得中药粉体;其中中药材包括枸杞、淫羊藿、红景天和丹参;按照质量比枸杞:淫羊藿:红景天:丹参为40:10:25:25;
(2)水溶功能性多糖溶液的制备:首先取200g步骤(1)所得中药粉体溶解于水中,中药粉体与水的质量比为3:7;溶解后加入由果胶酶、纤维素酶和淀粉酶按照质量比4:6:3复合而成的复合酶进行酶解,酶解时间为10h,酶解温度为40℃,pH值控制在3.5~5,复合酶的添加量为中药粉体加入量的8%,得到酶解液;然后将酶解液在离心转速为4000r/min条件下离心50min,取上清液,即得水溶功能性多糖溶液;
(3)水溶功能性多糖溶液的发酵处理:在步骤(2)所得的水溶功能性多糖溶液中加入活化后的复合乳酸菌进行发酵处理,发酵温度为37℃,发酵时间为12h,得到发酵液;复合乳酸菌的添加量为水溶功能性多糖溶液总质量的5%;复合乳酸菌为婴儿双歧杆菌、德氏乳杆菌和鼠李糖乳杆菌;
(4)复合凝聚体系的制备:将步骤(3)所得的发酵液与脂溶性活性成分和乳化剂加入至高速匀浆机,在匀浆转速为15000r/min条件下匀浆30min制得复合凝聚体系;所述脂溶性活性成分由维生素A、维生素D3、维生素E、维生素K2、人体必需矿物质、高活性氨基酸和ω-3-不饱和脂肪酸有机复配而成,且维生素A的添加量为发酵液质量的5%,维生素D3的添加量为发酵液质量的3%,维生素E的添加量为发酵液质量的8%,维生素K2的添加量为发酵液质量的4%,人体必需矿物质的添加量为发酵液质量的10%,高活性氨基酸的添加量为发酵液质量的15%,ω-3-不饱和脂肪酸的添加量为发酵液质量的12%;其中人体必需矿物质由葡萄糖酸镁、葡萄糖酸锌和葡萄糖酸铁按质量比2:4:3组成;高活性氨基酸由精氨酸、谷氨酸、甘氨酸和γ-氨基丁酸按质量比3:5:4:3组成;ω-3-不饱和脂肪酸由DHA和EPA按质量比4:7组成;所述乳化剂由二乙二醇脂肪酸酯、聚氧乙烯蓖麻油和聚氧乙烯植物油按质量比1:0.5:1.5组成,其中乳化剂的添加量为发酵液与脂溶性活性成分混合总量的2%,HLB值控制在12。
(5)微胶囊湿囊的制备:首先选用气流式粉碎机在粉碎压力为1.5MPa,转速为2000r/min条件下将蛋白进行超微粉碎处理,粉碎后过600目筛分离、细化,再与步骤(4)所得复合凝聚体系进行高压均质处理,均质压力为45MPa,均质时间为20min;然后室温下静置12h,制备得到均一的微胶囊分散液,离心除去上清液得到微胶囊湿囊;所述蛋白与复合凝聚体系的质量比为5:5;所述蛋白由大豆分离蛋白、乳清分离蛋白、β-乳球蛋白及β-伴大豆球蛋白按质量比1:1.5:1:0.5混合而成;
(6)专用营养补充剂的制备:对步骤(5)所得的微胶囊湿囊进行冷冻干燥,预冻温度为-50℃,物料厚度为5mm,冷冻干燥时间为35h,制得专用营养补充剂。
实施例3
所述改善大学生亚健康状态的专用营养补充剂制备方法,具体步骤如下:
(1)原料的破壁处理:对作为原料的中药材在气流压力为2.5MPa,转速为3000r/min条件下进行气流超微粉碎,粉碎后过600目筛制得中药粉体;其中中药材包括枸杞、淫羊藿、红景天和丹参;按照质量比枸杞:淫羊藿:红景天:丹参为43:8:22:27;
(2)水溶功能性多糖溶液的制备:首先取200g步骤(1)所得中药粉体溶解于水中,中药粉体与水的质量比为5:5;溶解后加入由果胶酶、纤维素酶和淀粉酶按照质量比5:8:4复合而成的复合酶进行酶解,酶解时间为9h,酶解温度为50℃,pH值控制在3.5~5,复合酶的添加量为中药粉体加入量的6%,得到酶解液;然后将酶解液在离心转速为4000r/min条件下离心35min,取上清液,即得水溶功能性多糖溶液;
(3)水溶功能性多糖溶液的发酵处理:在步骤(2)所得的水溶功能性多糖溶液中加入活化后的复合乳酸菌进行发酵处理,发酵温度为37℃,发酵时间为10h,得到发酵液;复合乳酸菌的添加量为水溶功能性多糖溶液总质量的3%;复合乳酸菌为嗜酸乳杆菌、保加利亚乳杆菌和鼠李糖乳杆菌;
(4)复合凝聚体系的制备:将步骤(3)所得的发酵液与脂溶性活性成分和乳化剂加入至高速匀浆机,在匀浆转速为12000r/min条件下匀浆30min制得复合凝聚体系;所述脂溶性活性成分由维生素A、维生素D3、维生素E、维生素K2、人体必需矿物质、高活性氨基酸和ω-3-不饱和脂肪酸有机复配而成,且维生素A的添加量为发酵液质量的3%,维生素D3的添加量为发酵液质量的2%,维生素E的添加量为发酵液质量的7%,维生素K2的添加量为发酵液质量的3%,人体必需矿物质的添加量为发酵液质量的9%,高活性氨基酸的添加量为发酵液质量的12%,ω-3-不饱和脂肪酸的添加量为发酵液质量的10%;其中人体必需矿物质由葡萄糖酸镁、葡萄糖酸锌和葡萄糖酸铁按质量比1:3:2组成;高活性氨基酸由精氨酸、谷氨酸、甘氨酸和γ-氨基丁酸按质量比2:3:4:2组成;ω-3-不饱和脂肪酸由DHA和EPA按质量比2:4组成;所述乳化剂由聚氧乙烯单月桂酸酯和失水山梨醇月桂酸酯按质量比2:1组成,其中乳化剂的添加量为发酵液与脂溶性活性成分混合总量的3.5%,HLB值控制在10。
(5)微胶囊湿囊的制备:首先选用气流式粉碎机在粉碎压力为1.2MPa,转速为2000r/min条件下将蛋白进行超微粉碎处理,粉碎后过400目筛分离、细化,再与步骤(4)所得复合凝聚体系进行高压均质处理,均质压力为30MPa,均质时间为15min;然后室温下静置10h,制备得到均一的微胶囊分散液,离心除去上清液,得到微胶囊湿囊;所述蛋白与复合凝聚体系的质量比为4:6;所述蛋白由大豆分离蛋白、乳清分离蛋白和β-伴大豆球蛋白按质量比3:1.5:1混合而成;
(6)专用营养补充剂的制备:对步骤(5)所得的微胶囊湿囊进行冷冻干燥,预冻温度为-40℃,物料厚度为10mm,冷冻干燥时间为30h,制得专用营养补充剂。
对比例1
在气流超微粉碎处理后的中药材水溶液中(枸杞:淫羊藿:红景天:丹参质量比=45:5:20:30,固液比为4:6),加入果胶酶、纤维素酶及淀粉酶所组成的混合酶(三者间质量比为为3:8:2,占中药粉末质量4%),在水浴搅拌的条件下进行复合酶解(温度控制在35±5℃),酶解8h后离心取上清液,加入脂溶性活性成分(添加量与实施例1中相同)并在高速匀浆过程中滴入1%的HLB值=9的乳化剂,以8000r/min的转速匀浆处理20min后得到复合凝聚体系。在高压均质机内将超微粉碎处理后的乳清分离蛋白、β-乳球蛋白及β-伴大豆球蛋白与所得到的复合凝聚体系(固液比为3:7)进行均质、静置处理后得到微胶囊湿囊,并于冷冻干燥机内冷冻处理获得大学生亚健康专用营养补充剂。该对比例并未进行复合乳酸菌发酵处理。
对比例2
在常规粉碎处理后的中药材中加入1.5倍质量的水溶液(枸杞:淫羊藿:红景天:丹参质量比=45:5:20:30),并在搅拌的条件下加入果胶酶、纤维素酶及淀粉酶所组成的混合酶(三者间质量比为为3:8:2,占中药粉末质量4%),于35±5℃反应8h后离心取上清。在已灭菌处理后上清液中接入保加利亚乳杆菌及婴儿双歧杆菌的混合菌液(菌液添加量为上清液总质量的3%)并于35℃密闭发酵8h,发酵完成后离心取上清。将上清液与脂溶性活性成分(添加量与实施例1中相同)充分混合后加入1%的HLB值=9的乳化剂,以8000r/min的转速高速匀浆处理20min后得到复合凝聚体系。将超微粉碎处理后的乳清分离蛋白、β-乳球蛋白及β-伴大豆球蛋白的混合粉(乳清分离蛋白:β-乳球蛋白:β-伴大豆球蛋白质量比为1.5:1:1)倒入复合凝聚体系内(固液比为3:7),通过高压均质机进行均质处理得到分散性良好的复合凝聚微胶囊分散液(均质压力为20MPa,均质时间为15min),并通过室温静置处理得到微胶囊湿囊。将湿囊置于已预冻处理过(预冻温度为-37℃)的真空冷冻干燥机内冷冻处理20h后得到大学生亚健康专用营养补充剂。该对比例并未对中药材进行气流超微粉碎。
试验例1
1、营养成分释放率实验:
(i)标准曲线的绘制
将所得到的大学生亚健康专用营养补充剂置于10倍质量的水中,在37.3±0.5℃的水浴锅中进行静置处理,10h后离心取上清液,利用液相色谱对上清液中的丹酚酸B活性成分含量进行检测,检测条件为以十八烷基硅烷键合硅胶为填充剂,以乙腈-0.1%磷酸溶液(22:78)为流动相,柱温为20℃,检验波长为286nm,流速为1.2mL/min。
根据高效液相色谱分析的结果,通过计算峰面积的大小,测定营养补充剂中丹酚酸B的含量。由图1中丹酚酸B的标准样品的液相谱图可以看出,丹酚酸B的出峰位置位于27.5min左右且峰型符合正态分布。同时,以丹酚酸B的进样量为横坐标,以峰面积为纵坐标进行标准曲线的绘制,由图2标准曲线可以知道丹酚酸B的标准曲线线性回归方程为y=106803x-41496,R2=0.9983,线性关系良好可用于后面营养补充剂中活性成分丹酚酸B释放率的测定。
(ii)大学生亚健康专用营养补充剂中活性成分释放率
由图3可以看出,实施例1丹酚酸B的峰面积远大于对比例1和对比例2,表明大学生亚健康专用营养补充剂制备过程中所选用的超微破碎处理、复合酶解及益生菌发酵对于营养补充剂中活性成分的释放具有积极的促进作用,且三种工艺的有机结合远比两种工艺随机结合所获得的活性成分的释放率高。
2、营养补充剂对小鼠肠道菌群的改善效果
(i)实验动物准备及样品采集
本发明选用200只雄性健康小鼠(体重25±5g),室内温度控制在24-25℃,保持良好通风,湿度控制在50-70%之间。适应饲养4天后,随机分为六组每组10只,将营养补充剂分散于蒸馏水中,分别配制浓度为15%、10%、5%、15%、15%的高剂量组、中剂量组、低剂量组、对比例1高剂量组和对比例2高剂量组,正常对照组每日灌胃相同体积的生理盐水。每日每组每只按0.15mL/10g进行灌胃,1次1天,连续灌胃5天后无菌采集小鼠新鲜粪便0.1-0.4g置于已称重灭菌过的小试管中进行称重,称重完成后加入10倍于粪便质量的无菌水并进行充分震荡。
(ii)肠道菌群数量测定
将稀释了9倍后的粪便水0.1mL分别接种于Lbs琼脂、BBL琼脂、EMB及胆汁七叶苷叠氮钠琼脂上进行肠道菌群中乳杆菌、双歧杆菌、肠杆菌及肠球菌的菌群数量测定,并按照各种菌对需养及厌氧条件的需求进行培养,培养结束后,观察并计数各类菌的菌落数。结果见表1。
由表1可以看出,小鼠在经高、中、低剂量营养补充剂灌胃处理后其肠道菌群数量中四种菌的数量与对照组间无显著性差异,但灌胃前后四种菌的数量均有所增加,益生菌的增加量相比于有害菌增加幅度较大,表明营养补充剂的摄入对改善肠道菌群结构具有积极作用。对比例高剂量组相比于高剂量组而言,改善效果不甚突出,但对于未经过发酵处理的对比例1来说,其有益菌增加幅度是所有分组中增加最少的,这也表明了营养补充剂制备工艺中的步骤3对于营养补充剂改善肠道菌群起着至关重要的作用。
3、营养补充剂对亚健康状态小鼠改善效果:
(ⅰ)实验动物准备
本发明选用150只4-5周龄的雌性小鼠(体重约为25±5g)为实验对象,实验开始前进行适应性游泳训练:将小鼠置于25±3℃的恒温水槽中进行训练,一周训练三次,每次5分钟,连续训练三周后进行体重检测并进行负重力竭游泳检测(负重量为自身体重的5%)并记录每次游泳时间。根据体重及游泳时间从中筛选出70只差异较小的进行后续实验。
(ii)实验动物分组及模型构建
将筛选出的70只小鼠禁食12h后,随机分为空白对照组、模型对照组及营养剂高、中、低剂量组,按照每组10只的量进行单笼喂养。造模期间,空白组正常饲养,其余小组在正常笼(6h)及水深为0.8cm的水笼(18h)中交替饲养,并连续7天进行力竭实验。造模过程中除正常饮食外,在小鼠进入水笼前1小时进行灌胃处理。空白组及模型对照组每次灌胃0.6mL纯净水,高、中、低剂量组、对比例1高剂量组及对比例2高剂量组分别灌胃浓度为4%,2%、1%、4%及4%的营养剂溶液0.6mL。7天后进行负重力竭游泳测试,并对体重和力竭游泳时间进行记录。
②检测指标
(i)行为学指标及体重变化
行为学方面,空白小组精神状态良好,皮毛有光泽且行动灵敏;模型对照组小鼠精神欠佳,易烦躁,皮毛无光泽,活动较少;高、中、低剂量组、对比例1高剂量组和对比例2高剂量组相比于模型对照组而言皮毛有光泽,反应能力及活动频率均有所提高,但后两者相对于前三者而言具有一定的差异性。
组别 | 数量/只 | 体重/g |
空白组 | 10 | 27.15±1.43 |
模型对照组 | 10 | 22.65±1.57 |
高剂量组 | 10 | 25.47±1.28 |
中剂量组 | 10 | 25.03±1.41 |
低剂量组 | 10 | 24.86±1.25 |
对比例1高剂量组 | 10 | 24.33±1.36 |
对比例2高剂量组 | 10 | 24.48±1.22 |
由表2可知,与空白组相比,造模各组体重均出现不同程度的下降,但相对于模型对照组而言,高、中、低剂量组体重变化具有显著性差异,表明营养剂的补充对维持小鼠体重具有较好的效果,对比例高剂量组虽然比模型对照组具有改善作用,但相比于高、中、低剂量组来说改善效果不大,该结果表明本发明中所描述的营养补充剂的功效与制备工艺具有很大的关系,工艺的改变将不利于亚健康状态的改善,效果差强人意。
(ii)力竭游泳时间比较
组别 | 数量/只 | 力竭游泳时间/s |
空白组 | 10 | 386±87.89 |
模型对照组 | 10 | 172±65.43 |
高剂量组 | 10 | 273±73.32 |
中剂量组 | 10 | 254±70.65 |
低剂量组 | 10 | 247±68.52 |
对比例1高剂量组 | 10 | 238±69.54 |
对比例2高剂量组 | 10 | 242±62.83 |
通过表3可以看出,与空白组相比,造模组力竭游泳时间大幅下降,差异性显著,其中模型对照组力竭游泳时间最短,小鼠疲劳程度较大,高、中、低剂量组相较于模型对照组具有显著改善,表明营养剂对改善疲劳具有良好功效,而对比例与高、中、低剂量间的差异性再次说明了该营养补充剂制备工艺与功效之间具有直接关系,不可任意更改。
表4实施例1-3及对比例1-2与市售产品的指标对比表
通过表4中的数据可以看出,本发明的产品与市售产品相比,其包埋率、PDI分布及肠液中活性成分的累积释放率均优于市售的ω-3-不饱和脂肪包埋微胶囊,且产率较高,表明该类方法所生产的营养补充剂效果显著,具有很好的市场推广价值。实施例1与对比例1、2之间的对比可以发现三者之间的差距并不显著,产生这种现象的原因可以归结于ω-3-不饱和脂肪酸属于脂溶性活性成分,其添加与超微破碎、复合酶解及益生菌发酵之间无直接关系,这也进一步表明超微破碎、复合酶解和益生菌发酵直接影响着亚健康营养补充剂的活性成分释放及肠道菌群结构改善,是本发明的关键核心技术。
由图4可知,市售产品中的ω-3不饱和脂肪酸在肠道呈现快速释放趋势,180min后释放率高达75%,而实施例1,2,3在肠道中的释放率有所延缓(以实施例2为最佳),表明本技术所制备的微胶囊相比市售产品而言具有有效物质缓慢释放的功效,产品时效增加,具有一定的市场竞争力。
Claims (5)
1.一种改善大学生亚健康状态的专用营养补充剂制备方法,包括以下步骤:
(1)原料的破壁处理:将作为原料的中药材进行破壁处理,制得中药粉体;其中所述中药材包括枸杞、淫羊藿、红景天和丹参;按照质量比枸杞:淫羊藿:红景天:丹参为35~45:4~10:20~30:25~30;
(2)水溶功能性多糖溶液的制备:首先将步骤(1)所得中药粉体溶解后加入复合酶进行酶解,得到酶解液;然后将酶解液离心取上清液,即得水溶功能性多糖溶液;
(3)水溶功能性多糖溶液的发酵处理:在步骤(2)所得的水溶功能性多糖溶液中加入活化后的复合乳酸菌进行发酵处理,得到发酵液;
(4)复合凝聚体系的制备:将步骤(3)所得的发酵液与脂溶性活性成分和乳化剂匀浆制得复合凝聚体系;
(5)微胶囊湿囊的制备:首先将超微处理后的蛋白与步骤(4)所得复合凝聚体系进行均质处理;然后静置,制备得到均一的微胶囊分散液,离心除去上清液得到微胶囊湿囊;
(6)专用营养补充剂的制备:对步骤(5)所得的微胶囊湿囊进行冷冻干燥,冷冻干燥的预冻温度为-50~-30℃,物料厚度为5~15mm,冷冻干燥时间为20~35h;制得专用营养补充剂;
所述步骤(2)中复合酶由果胶酶、纤维素酶和淀粉酶按照质量比3~5:6~9:2~4复合而成;
所述步骤(3)中复合乳酸菌为嗜酸乳杆菌、保加利亚乳杆菌、婴儿双歧杆菌、德氏乳杆菌和鼠李糖乳杆菌中的至少一种;
所述步骤(4)中脂溶性活性成分由维生素A、维生素D3、维生素E、维生素K2、人体必需矿物质、高活性氨基酸和ω-3-不饱和脂肪酸有机复配而成;其中人体必需矿物质由葡萄糖酸镁、葡萄糖酸锌和葡萄糖酸铁按质量比1~2:1~4:1~3组成;高活性氨基酸由精氨酸、谷氨酸、甘氨酸和γ-氨基丁酸按质量比1~3:3~5:2~4:1~3组成;所述乳化剂的添加量为发酵液与脂溶性活性成分混合总量的0.5%~3.5%,HLB值控制在9~12;
所述步骤(5)中超微处理选用气流式粉碎机,粉碎压力为0.7~1.5MPa,转速为1500~2000r/min,粉碎后过200~600目筛分离、细化;所述蛋白与复合凝聚体系的质量比为3~6:4~7;所述均质处理采用高压均质,均质压力为20~45MPa,均质时间为15~20min;所述静置的时间为8~12h;所述蛋白为大豆分离蛋白、乳清分离蛋白、β-乳球蛋白及β-伴大豆球蛋白中的至少一种。
2.根据权利要求1所述改善大学生亚健康状态的专用营养补充剂制备方法,其特征在于,所述步骤(1)中破壁处理采用气流超微粉碎,其中气流超微粉碎的气流压力为2~3MPa,转速为2500~4000r/min;粉碎后过400~600目筛。
3.根据权利要求1所述改善大学生亚健康状态的专用营养补充剂制备方法,其特征在于,所述步骤(2)将中药粉体溶解于水中,中药粉体与水的质量比为3~5:5~7;所述复合酶的添加量为中药粉体加入量的2~8%;所述酶解的酶解时间为8~10h,酶解温度为35~50℃,pH值控制在3 .5~5;所述离心的离心转速为2000~4000r/min,离心时间为30~50min。
4.根据权利要求1所述改善大学生亚健康状态的专用营养补充剂制备方法,其特征在于,所述步骤(3)中复合乳酸菌的添加量为水溶功能性多糖溶液总质量的2%~5%;发酵温度为30~39℃,发酵时间为8h~12h。
5.根据权利要求1所述改善大学生亚健康状态的专用营养补充剂制备方法,其特征在于,所述步骤(4)中匀浆采用高速匀浆机,匀浆转速为8000~5000r/min,匀浆时间为20~30min;所述维生素A的添加量为发酵液质量的3%~5%,维生素D3的添加量为发酵液质量的1%~3%,维生素E的添加量为发酵液质量的5%~8%,维生素K2的添加量为发酵液质量的2%~4%,人体必需矿物质的添加量为发酵液质量的6%~10%,高活性氨基酸的添加量为发酵液质量的8%~15%,ω-3-不饱和脂肪酸的添加量为发酵液质量的6%~12%;ω-3-不饱和脂肪酸由DHA和EPA按质量比2~4:3~7组成;所述乳化剂为聚氧乙烯氧丙烯油酸酯、聚氧乙烯单月桂酸酯、失水山梨醇月桂酸酯、二乙二醇脂肪酸酯、聚氧乙烯蓖麻油和聚氧乙烯植物油中的至少一种。
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