CN111990651A - 一种含多糖的鱼粉含片及其制备方法 - Google Patents
一种含多糖的鱼粉含片及其制备方法 Download PDFInfo
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- CN111990651A CN111990651A CN202010864131.XA CN202010864131A CN111990651A CN 111990651 A CN111990651 A CN 111990651A CN 202010864131 A CN202010864131 A CN 202010864131A CN 111990651 A CN111990651 A CN 111990651A
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- fish
- buccal tablet
- mass
- fish meal
- snakehead
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Abstract
本发明公开了一种含多糖的鱼粉含片及其制备方法,属于食品加工技术领域。本发明提供了一种鱼粉含片,此鱼粉含片中添加有岩藻多糖和/或海参多糖,并且,由于岩藻多糖和/或海参多糖是在压片前与干燥颗粒混合,在此鱼粉含片的制备过程中,岩藻多糖和/或海参多糖的活性不会消失,因此,此鱼粉含片可预防和治疗新型冠状病毒感染。
Description
技术领域
本发明涉及一种含多糖的鱼粉含片及其制备方法,属于食品加工技术领域。
背景技术
新型冠状病毒(2019-nCoV),正式分类命名为严重急性呼吸综合征冠状病毒2(Severe acute respiratory syndrome coronavirus 2,SARS-CoV-2),简称新冠病毒(novel coronavirus),于2019年底开始出现,在全球迅速传播扩散,已造成上百万人感染。人感染了冠状病毒后常见体征有呼吸道症状、发热、咳嗽、气促和呼吸困难等。在较严重病例中,感染可导致肺炎、严重急性呼吸综合征、肾衰竭,甚至死亡。因此,急需研发可预防和治疗新型冠状病毒感染的产品。
发明内容
[技术问题]
本发明要解决的技术问题是提供一种可预防和治疗新型冠状病毒感染的产品。
[技术方案]
为解决本发明的技术问题,本发明提供了一种制备鱼粉含片的方法,所述方法为在制备鱼粉含片的过程中添加活性物质;所述活性物质为岩藻多糖和/或海参多糖。
在本发明的一种实施方式中,所述方法为取黑鱼整鱼,将黑鱼宰杀,得到鱼身;将鱼身浸泡于酵母溶液中去腥后清洗干净,得到去腥鱼身;采集去腥鱼身上的肉,得到鱼骨和鱼肉;在鱼肉中添加水后,将鱼肉搅碎成糜状,得到鱼肉浆;将鱼肉浆、蛋白酶和水混合后进行酶解,得到酶解产物;将酶解产物过滤,得到酶解液;将酶解液第一次干燥,得到黑鱼蛋白粉;将鱼骨蒸煮后第二次干燥、粉碎,得到黑鱼骨粉;将黑鱼蛋白粉、黑鱼骨粉和填充剂、矫味剂、香精、粘合剂、湿润剂混合,得到软材;将软材造粒,得干燥颗粒;将干燥颗粒和活性物质混合后(用压片机)进行压片,得到鱼粉含片。
在本发明的一种实施方式中,所述宰杀包括去鳞、放血、去鳃、去内脏、去皮、洗净。
在本发明的一种实施方式中,所述酵母溶液由活性干酵母粉与水混合而得。
在本发明的一种实施方式中,所述酵母溶液中,活性干酵母粉的浓度为5~15g/L。
在本发明的一种实施方式中,所述浸泡的时间为0.5~1.5h。
在本发明的一种实施方式中,所述水在鱼肉中的添加量占鱼肉总质量的20~30%。
在本发明的一种实施方式中,所述蛋白酶为木瓜蛋白酶和风味蛋白酶。
在本发明的一种实施方式中,所述木瓜蛋白酶和风味蛋白酶的质量比为1:2~4。
在本发明的一种实施方式中,所述将鱼肉浆、蛋白酶和水混合为先将蛋白酶添加至鱼肉浆中,得到酶解体系,然后将水添加至酶解体系中至酶解体系的固液比为1:2~4。
在本发明的一种实施方式中,所述蛋白酶在鱼肉浆中的添加量占鱼肉浆总质量的0.5~1.5%。
在本发明的一种实施方式中,所述酶解的温度为50℃、pH为6.5、时间为4h。
在本发明的一种实施方式中,所述第一次干燥为喷雾干燥。
在本发明的一种实施方式中,所述喷雾干燥的进料浓度为20~25%、进风温度为170~180℃、进料速度为20~30mL/min。
在本发明的一种实施方式中,所述蒸煮为用高温高压蒸煮锅蒸煮。
在本发明的一种实施方式中,所述蒸煮的温度为121℃、时间为15~20min。
在本发明的一种实施方式中,所述第二次干燥为先用烤箱烤制,再用烘箱烘干。
在本发明的一种实施方式中,所述烤制的温度为120~150℃、时间为5~10min。
在本发明的一种实施方式中,所述烘干的温度为50~60℃。
在本发明的一种实施方式中,所述粉碎为用超微粉碎机粉碎。
在本发明的一种实施方式中,所述填充剂为β-环状糊精;所述矫味剂为针叶樱桃粉和罗汉果糖;所述香精为薄荷香精;所述粘合剂为羟丙基甲基纤维素;所述湿润剂为乙醇;所述润滑剂为硬脂酸镁。
在本发明的一种实施方式中,所述湿润剂为体积分数40%的乙醇。
在本发明的一种实施方式中,以质量计,所述软材含有黑鱼蛋白粉15~45%、黑鱼骨粉5~25%、β-环状糊精15%~35%、针叶樱桃粉1~5%、罗汉果糖0.3%~1%、薄荷香精0.05~0.15%、羟丙基甲基纤维素1%~10%、乙醇20~40%、硬脂酸镁0.5~2%。
在本发明的一种实施方式中,所述造粒包括干燥、研磨、过筛。
在本发明的一种实施方式中,所述活性物质在干燥颗粒中的添加量占干燥颗粒总质量的0.05~0.2%。
在本发明的一种实施方式中,所述压片为用压片机压片。
本发明还提供了一种鱼粉含片,所述鱼粉含片为使用上述方法制备而得。
[有益效果]
(1)本发明提供了一种鱼粉含片,此鱼粉含片中添加有岩藻多糖和/或海参多糖,并且,由于岩藻多糖和/或海参多糖是在压片前与干燥颗粒混合,在此鱼粉含片的制备过程中,岩藻多糖和/或海参多糖的活性不会消失,因此,此鱼粉含片可预防和治疗新型冠状病毒感染。
(2)本发明提供了一种鱼粉含片,此鱼粉含片的原材料为黑鱼,黑鱼肉质细嫩,营养丰富,具有很好的营养和药用价值,因此,此鱼粉含片可起到营养强化作用。
(3)本发明提供了一种鱼粉含片,此鱼粉含片的制备同时使用了鱼肉和鱼骨,有利于提高资源利用率,充分发挥原料营养价值。
附图说明
图1:不同浓度的泡叶藻岩藻多糖对新冠病毒的抑制作用,横坐标:泡叶藻岩藻多糖的浓度,纵坐标:感染细胞率(%)=进入细胞的病毒数量/总病毒数量×100。
图2:将SARS-CoV-2病毒与不同浓度的岩藻多糖混合处理后的细胞免疫荧光图。岩藻多糖的浓度为500、250、125、62.5、31.3、15.6μg/mL。阴性对照为空白培养基。
图3:将SARS-CoV-2病毒与不同浓度的海参多糖的混合处理后的细胞免疫荧光图。海参多糖的终浓度为500、250、125、62.5、31.3、15.6、7.8、3.9μg/mL。阴性对照为空白培养基。
具体实施方式
下面结合具体实施例对本发明进行进一步的阐述。
下述实施例中涉及的罗汉果糖购自湖南华城生物资源股份有限公司;下述实施例中涉及的针叶樱桃粉购自陕西浩洋生物科技有限公司;下述实施例中涉及的硬脂酸镁购自广东华盛食品配料公司;下述实施例中涉及的β-环状糊精购自河南华悦化工产品有限公司;下述实施例中涉及的乙醇和羟甲基纤维素购自天津大茂化学试剂厂;下述实施例中涉及的薄荷香精购自成都万象宏润生物科技有限公司;下述实施例中涉及的木瓜蛋白酶和风味蛋白酶购自诺维信(中国)生物技术有限公司。
实施例1-1:制备泡叶藻(褐藻)岩藻多糖
泡叶藻岩藻多糖的制备方法如下:
S1、将泡叶藻(Ascophyllum nodosum)洗净、沥干、自然风干、粉碎并过80目筛,得泡叶藻粉A;
S2、将步骤S1所述泡叶藻粉A置于25℃无水乙醇中浸泡4h,纱布过滤取沉淀A,将所述沉淀A置于25℃无水乙醇中搅拌4h,纱布过滤取沉淀B,将所述沉淀B置于25℃无水乙醇中浸泡4h,纱布过滤取沉淀C室温晾干,从而除去脂类及脂溶性小分子,得泡叶藻粉B;其中,所述泡叶藻粉A、沉淀B和本步骤所述无水乙醇的重量体积比均为1:4g/mL;
S3、取步骤S2所述泡叶藻粉B,加入pH=5的磷酸氢二钠-柠檬酸缓冲液、纤维素酶、果胶酶和木瓜蛋白酶,搅拌混匀,50℃水浴振荡酶解4h,使岩藻多糖解离,后加热至98℃,保持10分钟以钝化酶活,所得混合物于4500r/min室温离心15min,取上清液;其中,所述泡叶藻粉B和所述磷酸氢二钠-柠檬酸缓冲液的重量体积比为1:30g/mL;所述泡叶藻粉B、纤维素酶(比酶活100units/mg)、果胶酶(比酶活50units/mg)和木瓜蛋白酶(比酶活2units/mg)的重量比为12500:42:6:6;
S4、向步骤S3所述上清液中边搅拌边加入过量的CaCl2,于4500r/min室温离心15min,将褐藻胶沉淀除去,取上清液;使用的步骤S3的上清液和CaCl2体积重量比是20:1mL/g;
S5、向步骤S4所述上清液中加入十六烷基三甲基溴化铵(CTAB),使岩藻多糖沉淀,将所得混合物在4500r/min室温离心15min收集沉淀,将沉淀溶解于3mol/L CaCl2溶液中,再加无水乙醇,4℃放置24h,使岩藻多糖沉淀,4500r/min、4℃离心15min,收集沉淀;使用的步骤S4的上清液和CTAB体积重量比是50:1mL/g;所述沉淀和所述3mol/L CaCl2溶液的重量体积比是1:3g/mL;所述CaCl2溶液和所述无水乙醇的体积比为2:3;
S6、将步骤S5所述沉淀用体积分数为80%的乙醇洗涤3次,再使用体积分数为95%乙醇洗沉淀3次,室温晾干,超纯水溶解,使用分子量3500Da的透析袋自来水流水透析24h,然后使用超纯水作为透析液透析48h,除去岩藻多糖中含有的氯化钙及其它盐离子,其中每2h换透析液,在真空度为1pa和温度为-60℃的条件下冻干72h,得到泡叶藻岩藻多糖(ANP);所述沉淀和所述体积分数为80%的乙醇溶液的重量体积比是1:3g/mL;所述沉淀和所述体积分数为95%的乙醇溶液的重量体积比是1:3g/mL;所述沉淀和所述超纯水的重量体积比是1:150g/mL。
本实施例还可以包括溶液配制、超纯水的制备等前处理步骤。
实施例1-2:确定实施例1-1所制备的泡叶藻岩藻多糖的结构特性和组成
具体方法如下:
采用明胶比浊法进行泡叶藻岩藻多糖硫酸基含量的测定;
采用BCA法进行泡叶藻岩藻多糖蛋白含量的测定;
采用间羟基联苯法进行泡叶藻岩藻多糖糖醛酸含量的测定;
采用苯酚硫酸法进行泡叶藻岩藻多糖总糖含量的测定;
采用凝胶渗透色谱法进行泡叶藻岩藻多糖分子量的测定;
采用高效液相色谱+PMP衍生化法进行泡叶藻岩藻多糖单糖组成的测定;
采用傅里叶红外光谱法进行泡叶藻岩藻多糖官能团的测定。
结果表明,泡叶藻岩藻多糖的分子量为490kDa;糖醛酸含量为2.9~3.2%;蛋白含量为3.8~4.0%;硫酸基含量为28~30%;总糖含量为54%;所述褐藻岩藻多糖的单糖组成为:摩尔比为6.5:1.1:1的岩藻糖、甘露糖和半乳糖;官能团包括羟基、羧基和硫酸基等官能团。
实施例1-3:验证泡叶藻岩藻多糖阻止SARS-CoV-2病毒侵入机体细胞的用途
将编码HCoV-19刺突蛋白的基因的全长序列克隆到pCAGGS载体用于假病毒的生产,所得到的重组载体称为pCAGGS-HCoV-19-S。通过DNA测序确认pCAGGS-HCoV-19-S构建成功。将pCAGGS-HCoV-19-S和pNL4-3的质粒共转染到HEK 293T细胞,培养48h后,收集含有SARS-CoV-2假病毒模型的上清液,并通过感染Huh7细胞确定假病毒的50%组织细胞感染量(TCID50)。
使用该SARS-CoV-2假病毒模型评价泡叶藻岩藻多糖的抗新型冠状病毒的作用,具体步骤如下:
(1)选择生长状态良好的Huh7细胞,胰酶消化后,96孔铺板,培养过夜,至18-24h时细胞达到80-100%;
(2)每孔100TCID50假病毒,与含有泡叶藻岩藻多糖的无血清培养基混合,混合后泡叶藻岩藻多糖的终浓度为0.01mg/mL、0.1mg/mL和1mg/mL,37℃孵育30min。EK1肽作为阳性对照,空白无血清培养基作为阴性对照。
(3)以PBS洗涤Huh7细胞去除血清后,用3倍倍比稀释病毒与泡叶藻岩藻多糖的混合物感染Huh7细胞,每个孔100μL,每个样品设置三个平行孔,4-6h后,补加含有5%FBS血清的培养基100μL。
(4)48h测定Luciferase值。参考Promega公司Luciferase Assay SystemProtocol或Dual Luciferase Reporter Assay System Protocol。具体操作:倒扣96孔板,用PBS洗2遍,确保吸干PBS,然后加入30μL的裂解液,常温裂解30min,吸出10μL于白板上,底物50μL,测定luciferase值。
如下表1所示,泡叶藻岩藻多糖在浓度为0.01mg/mL、0.1mg/mL和1mg/mL时,都能够有效的抑制SARS-CoV-2病毒感染细胞。
表1不同浓度下的泡叶藻岩藻多糖对SARS-CoV-2病毒感染细胞的抑制作用
又进一步采用相同实验方法检测了多个浓度的泡叶藻岩藻多糖对新冠病毒的抑制作用,计算了IC50值。结果如图1所示,泡叶藻岩藻多糖抑制新冠病毒的IC50为0.327mg/mL。
而且,由于所使用的模型为仅具有S蛋白的SARS-CoV-2假病毒,可以推出泡叶藻岩藻多糖的作用靶点为S蛋白。
实施例1-4:购买的商品化岩藻多糖的组成分析与抗新冠病毒研究
参考中华人民共和国水产行业标准SC/T 3404-2012,测得青岛明月海藻集团有限公司的商品化岩藻多糖中硫酸基含量为28.5±0.1%,总糖含量为63.2±2.6%,岩藻糖含量为36.9±3.8%。
将SARS-CoV-2真病毒(来源于第二军医大学)和岩藻多糖用含有5%胎牛血清的DMEM培养基混匀,37℃放置1h,加入提前12h接种Vero E6细胞的96孔板(加入以前吸除原细胞培养液),培养24h,然后用免疫荧光检测病毒蛋白,DAPI染色细胞核。采用免疫荧光显微镜观测了终浓度为500、250、125、62.5、31.3、15.6、7.8μg/mL的岩藻多糖对病毒的抑制作用,结果如图2所示,在浓度范围15.6~500μg/mL浓度范围内,岩藻多糖均能够显著抑制新冠病毒对细胞的感染。
实施例2-1:制备海参多糖的方法
具体方法如下:
将海参(刺参,Stichopusjaponicus)洗净、水煮、沥干、剪成小块,然后冻干。将冻干样品置于4℃丙酮中浸泡24h,室温下晾干。以1g冻干样品为例,加入30mL 0.1mol/L乙酸钠缓冲溶液(pH 6.0)、100mg木瓜蛋白酶(比酶活2units/mg)、48mg乙二胺四乙酸和18mg半胱氨酸,涡旋混合,于60℃水浴振荡酶解24h,将反应混合物离心(6000g,15min,室温),取上清液。向上清液中加入1.6mL 10%氯化十六烷基吡啶溶液,室温下放置24h后,离心(8000g,15min,室温)取沉淀。将沉淀溶解于15mL 3mol/LNaCl-乙醇(100:15v/v))溶液中,再加入30mL 95%乙醇溶液,4℃放置24h,离心(8000g,15min,室温)取沉淀。将沉淀用10mL 80%乙醇洗2至3次后,再用10mL 95%乙醇洗2至3次,室温晾干,蒸馏水溶解,用透析袋(3500Da)进行除盐,冻干,得海参多糖。
本实施例还可以包括溶液配制、超纯水的制备等前处理步骤。
实施例2-2:确定实施例2-1所制备的海参多糖的结构特性和组成
具体方法如下:
采用1H NMR进行海参多糖结构特性和纯度检测;
采用凝胶渗透色谱法进行海参多糖分子量检测;
采用明胶比浊法进行海参多糖硫酸根含量检测;
采用高效液相色谱+PMP衍生化法进行海参多糖单糖组成检测;
采用傅里叶红外进行海参多糖官能团检测。
结果表明,海参多糖含有岩藻聚糖硫酸酯和岩藻糖基化硫酸软骨素,岩藻聚糖硫酸酯分子量>670kDa,岩藻糖基化硫酸软骨素的分子量>179kDa;硫酸根含量为26-28%;岩藻糖、葡萄糖醛酸和氨基半乳糖的摩尔比为9:0.8:1。
实施例2-3:使用假病毒模型评价海参多糖的抗新型冠状病毒的作用
将编码HCoV-19刺突蛋白的基因的全长序列克隆到pCAGGS载体用于假病毒的生产,构建得到的重组载体称为pCAGGS-HCoV-19-S。通过DNA测序确认pCAGGS-HCoV-19-S构建成功。将pCAGGS-HCoV-19-S和pNL4-3质粒共转染到HEK 293T细胞,培养48h后,收集含有SARS-CoV-2假病毒的上清液,并通过感染Huh7细胞确定假病毒的50%组织细胞感染量(TCID50)。
使用该SARS-CoV-2假病毒模型评价海参多糖的抗新型冠状病毒的作用,具体步骤如下:
(1)选择生长状态良好的Huh7细胞,胰酶消化后,96孔铺板,培养过夜,至18-24h时细胞达到80-100%;
(2)每孔100TCID50假病毒,与含有海参多糖的无血清培养基混合,混合后海参多糖的终浓度为0.01mg/mL、0.1mg/mL和1mg/mL,于37℃孵育30min。EK1肽作为阳性对照,空白无血清培养基作为阴性对照。
(3)以PBS洗涤Huh7细胞去除血清后,用3倍倍比稀释病毒与海参多糖的混合物并感染Huh7细胞,每个孔100μL,每个样品设置三个平行孔,4-6h后,补加含有5%FBS血清的培养基100μL。
(4)48h测定Luciferase值。参考Promega公司Luciferase Assay SystemProtocol或Dual Luciferase Reporter Assay System Protocol。具体操作:倒扣96孔板,用PBS洗2遍,确保吸干PBS,然后加入30μL的裂解液,常温裂解30min,吸出10μL于白板上,底物50μL,测定luciferase值,结果如下表2所示。
如下表2所示,海参多糖终浓度为0.1mg/mL和1mg/mL时,都能够有效的抑制SARS-CoV-2病毒进入细胞。而且由于所使用的模型为具有S蛋白的SARS-CoV-2假病毒,可以推出海参多糖的作用靶点为S蛋白。
表2不同浓度下的海参多糖对SARS-CoV-2病毒感染细胞的抑制作用
实施例2-4:海参多糖对duSARS-CoV-2真病毒的作用
SARS-CoV-2真病毒(来源于第二军医大学)和海参多糖用含有5%胎牛血清的DMEM培养基混匀,使海参多糖的终浓度为500、250、125、62.5、31.3、15.6、7.8、3.9μg/mL,37℃放置1h,加入提前12h接种Vero E6细胞的96孔板(加入以前吸除原细胞培养液),培养24h,然后用免疫荧光检测病毒蛋白,DAPI染色细胞核。用免疫荧光显微镜观测了500、250、125、62.5、31.3、15.6、7.8、3.9μg/mL八个浓度梯度的海参多糖对病毒的抑制作用,结果如图3所示,在500~3.9μg/mL浓度范围内,海参多糖均能够显著抑制新冠病毒对细胞的感染。
实施例3-1:鱼粉含片的制备
包括如下步骤:
取黑鱼整鱼,将黑鱼宰杀,得到鱼身;将鱼身浸泡于酵母溶液中去腥后清洗干净,得到去腥鱼身;采集去腥鱼身上的肉,得到鱼骨和鱼肉;在鱼肉中添加水后,将鱼肉搅碎成糜状,得到鱼肉浆;将鱼肉浆、蛋白酶和水混合后进行酶解,得到酶解产物;将酶解产物过滤,得到酶解液;将酶解液第一次干燥,得到黑鱼蛋白粉;将鱼骨蒸煮后第二次干燥、粉碎,得到黑鱼骨粉;将黑鱼蛋白粉、黑鱼骨粉和填充剂、矫味剂、香精、粘合剂、湿润剂混合,得到软材;将软材造粒,得干燥颗粒;将干燥颗粒和活性物质混合后(用压片机)进行压片,得到鱼粉含片;其中,所述宰杀包括去鳞、放血、去鳃、去内脏、去皮、洗净;所述酵母溶液由活性干酵母粉与水混合而得;所述酵母溶液中,活性干酵母粉的浓度为5~15g/L;所述浸泡的时间为0.5~1.5h;所述水在鱼肉中的添加量占鱼肉总质量的20~30%;所述蛋白酶为木瓜蛋白酶和风味蛋白酶;所述木瓜蛋白酶和风味蛋白酶的质量比为1:2~4;所述将鱼肉浆、蛋白酶和水混合为先将蛋白酶添加至鱼肉浆中,得到酶解体系,然后将水添加至酶解体系中至酶解体系的固液比为1:2~4;所述蛋白酶在鱼肉浆中的添加量占鱼肉浆总质量的0.5~1.5%;所述酶解的温度为50℃、pH为6.5、时间为4h;所述第一次干燥为喷雾干燥;所述喷雾干燥的进料浓度为20~25%、进风温度为170~180℃、进料速度为20~30mL/min;所述蒸煮为用高温高压蒸煮锅蒸煮;所述蒸煮的温度为121℃、时间为15~20min;所述第二次干燥为先用烤箱烤制,再用烘箱烘干;所述烤制的温度为120~150℃、时间为5~10min;所述烘干的温度为50~60℃;所述粉碎为用超微粉碎机粉碎;所述填充剂为β-环状糊精;所述矫味剂为针叶樱桃粉和罗汉果糖;所述香精为薄荷香精;所述粘合剂为羟丙基甲基纤维素;所述湿润剂为40%(v/v)的乙醇;所述润滑剂为硬脂酸镁;以质量计,所述软材含有黑鱼蛋白粉15~45%、黑鱼骨粉5~25%、β-环状糊精15%~35%、针叶樱桃粉1~5%、罗汉果糖0.3%~1%、薄荷香精0.05~0.15%、羟丙基甲基纤维素1%~10%、乙醇20~40%、硬脂酸镁0.5~2%;所述造粒包括干燥、研磨、过筛;所述活性物质在干燥颗粒中的添加量占干燥颗粒总质量的0.05~0.2%;所述压片为用压片机压片;所述活性物质为岩藻多糖和/或海参多糖(岩藻多糖的制备方法见实施例1-1或购自青岛明月海藻集团有限公司,海参多糖的制备方法见实施例2-1)。
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (10)
1.一种制备鱼粉含片的方法,其特征在于,所述方法为在制备鱼粉含片的过程中添加活性物质;所述活性物质为岩藻多糖和/或海参多糖。
2.如权利要求1所述的一种制备鱼粉含片的方法,其特征在于,所述方法为取黑鱼整鱼,将黑鱼宰杀,得到鱼身;将鱼身浸泡于酵母溶液中去腥后清洗干净,得到去腥鱼身;采集去腥鱼身上的肉,得到鱼骨和鱼肉;在鱼肉中添加水后,将鱼肉搅碎成糜状,得到鱼肉浆;将鱼肉浆、蛋白酶和水混合后进行酶解,得到酶解产物;将酶解产物过滤,得到酶解液;将酶解液第一次干燥,得到黑鱼蛋白粉;将鱼骨蒸煮后第二次干燥、粉碎,得到黑鱼骨粉;将黑鱼蛋白粉、黑鱼骨粉和填充剂、矫味剂、香精、粘合剂、湿润剂混合,得到软材;将软材造粒,得干燥颗粒;将干燥颗粒和活性物质混合后(用压片机)进行压片,得到鱼粉含片。
3.如权利要求2所述的一种制备鱼粉含片的方法,其特征在于,所述酵母溶液由活性干酵母粉与水混合而得。
4.如权利要求3所述的一种制备鱼粉含片的方法,其特征在于,所述酵母溶液中,活性干酵母粉的浓度为5~15g/L。
5.如权利要求2-4任一项所述的一种制备鱼粉含片的方法,其特征在于,所述将鱼肉浆、蛋白酶和水混合为先将蛋白酶添加至鱼肉浆中,得到酶解体系,然后将水添加至酶解体系中至酶解体系的固液比为1:2~4。
6.如权利要求5所述的一种制备鱼粉含片的方法,其特征在于,所述蛋白酶在鱼肉浆中的添加量占鱼肉浆总质量的0.5~1.5%。
7.如权利要求2-6任一项所述的一种制备鱼粉含片的方法,其特征在于,所述填充剂为β-环状糊精;所述矫味剂为针叶樱桃粉和罗汉果糖;所述香精为薄荷香精;所述粘合剂为羟丙基甲基纤维素;所述湿润剂为乙醇;所述润滑剂为硬脂酸镁。
8.如权利要求7所述的一种制备鱼粉含片的方法,其特征在于,以质量计,所述软材含有黑鱼蛋白粉15~45%、黑鱼骨粉5~25%、β-环状糊精15%~35%、针叶樱桃粉1~5%、罗汉果糖0.3%~1%、薄荷香精0.05~0.15%、羟丙基甲基纤维素1%~10%、乙醇20~40%、硬脂酸镁0.5~2%。
9.如权利要求2-8任一项所述的一种制备鱼粉含片的方法,其特征在于,所述活性物质在干燥颗粒中的添加量占干燥颗粒总质量的0.05~0.2%。
10.一种鱼粉含片,其特征在于,所述鱼粉含片为使用权利要求1-9任一所述的方法制备而得。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101756286A (zh) * | 2010-02-04 | 2010-06-30 | 林峰亮 | 一种鳄鱼口含片及其制备方法 |
CN103637245A (zh) * | 2013-12-02 | 2014-03-19 | 华中农业大学 | 一种补钙降血压的鱼蛋白肽钙咀嚼片及其制备方法 |
US20170267746A1 (en) * | 2016-03-16 | 2017-09-21 | Eric Hanxiang Sun | Techniques of preparing collagen active peptides |
CN107495184A (zh) * | 2017-07-31 | 2017-12-22 | 磐安县派普特生物科技有限公司 | 一种鱼骨粉和蛋白肽混合物咀嚼片 |
CN107647385A (zh) * | 2017-07-31 | 2018-02-02 | 浦江县泰如食品科技有限公司 | 一种鱼骨粉和蛋白肽混合物咀嚼片的制备方法 |
CN111471088A (zh) * | 2020-04-21 | 2020-07-31 | 北京中科微盾生物科技有限责任公司 | 一种抑制sars-cov-2感染的多肽、组合物及其用途 |
-
2020
- 2020-08-25 CN CN202010864131.XA patent/CN111990651A/zh active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101756286A (zh) * | 2010-02-04 | 2010-06-30 | 林峰亮 | 一种鳄鱼口含片及其制备方法 |
CN103637245A (zh) * | 2013-12-02 | 2014-03-19 | 华中农业大学 | 一种补钙降血压的鱼蛋白肽钙咀嚼片及其制备方法 |
US20170267746A1 (en) * | 2016-03-16 | 2017-09-21 | Eric Hanxiang Sun | Techniques of preparing collagen active peptides |
CN107495184A (zh) * | 2017-07-31 | 2017-12-22 | 磐安县派普特生物科技有限公司 | 一种鱼骨粉和蛋白肽混合物咀嚼片 |
CN107647385A (zh) * | 2017-07-31 | 2018-02-02 | 浦江县泰如食品科技有限公司 | 一种鱼骨粉和蛋白肽混合物咀嚼片的制备方法 |
CN111471088A (zh) * | 2020-04-21 | 2020-07-31 | 北京中科微盾生物科技有限责任公司 | 一种抑制sars-cov-2感染的多肽、组合物及其用途 |
Non-Patent Citations (2)
Title |
---|
叶云花等: ""淡水鱼的深加工与综合利用研究"", 《食品科技》, no. 4, pages 29 - 31 * |
程莉双等: ""利用鲤鱼骨生产富钙咀嚼片的配方及工艺研究"", 《天津农学院学报》, vol. 20, no. 2, pages 149 - 29 * |
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