CN108013471A - 牡蛎多糖复配物的制备方法 - Google Patents
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种牡蛎多糖的提取方法,其具体步骤为:将牡蛎鲜肉脱脂制;将脱脂牡蛎粉沸水浸提,过滤,离心,上清液干燥得水提取粗多糖;将水提沉淀物中加入碱液提取,离心,上清液冻干得碱提取粗多糖;向粗多糖中加水溶解,依次加入胰蛋白酶、胃蛋白酶反应,离心,上清液加入Sevage试剂脱蛋白,上层多糖溶液离心后取沉淀,冻干,即得牡蛎多糖;将水提取牡蛎多糖和碱提牡蛎多糖进行复配混合,然后制成胶囊剂型,即得牡蛎多糖复配物。有益效果为:本发明制备方法简单可行,机械化操作成度高,资源利用率高,能耗低,易于工业化规模生产;该复配物对肝损伤具有保护作用,具有改善心脏机能、促进血液循环、增强机体免疫力等效果。
Description
技术领域
本发明涉及天然产物深加工技术领域,特别涉及牡蛎多糖复配物的制备方法。
背景技术
牡蛎(oyster)是属于软体动物门双壳纲珍珠贝目的曲蛎科和牡蛎科(Ostreidae,真牡蛎)或燕蛤科(Aviculidae,珍珠牡蛎)的双壳类软体动物,又名蚝、海蛎子、蛎黄、蛎蛤或牡蛤。牡蛎是世界上第一大养殖贝类,在世界范围内已发现的牡蛎品种有100种左右,牡蛎也是我国重要海洋贝类,在我国沿海分布的牡蛎有20多种,主要有近江牡蛎、大连湾牡蛎、太平洋牡蛎(长牡蛎)(Crassostrea gigas)及密鳞牡蛎。我国是牡蛎养殖大国,牡蛎资源极为丰富。牡蛎又富含蛋白质、多糖、牛磺酸、不饱和脂肪酸、矿物质成分等生理活性物质,这些已经被开发为牡蛎功能食品,相关产品在市场上深受消费者的欢迎。还有很多学者利用食品科学、营养学、中西医学、生物学等学科的理论与技术深入研究牡蛎中活性物质的结构及其功能,继续开发新的牡蛎功能性产品。因此,开发牡蛎具有广阔的市场前景。牡蛎中多糖占干重的20%-40%,是产生免疫调节作用的活性物质。研究发现牡蛎多糖具有明显的抗凝血、降血脂、抗血栓、防治心血管疾病、提高机体免疫功能以及抗白细胞降低等生物活性。因此,牡蛎多糖成为保健食品领域研究开发的一个重点。
现有技术如授权公告号为CN 102406205 B的中国发明专利,公开了牡蛎多糖果汁饮料,该饮料以牡蛎多糖为主,辅以大鲵低聚糖肽、黑加仑、越橘、柠檬酸和果胶,具有降血压、保肝、改善睡眠、抗辐射损伤的作用,按质量百分比计有效成分如下:牡蛎多糖0.1%~10%、大鲵低聚糖肽0.1%~10%、黑加仑冻干粉0.1%~10%、越橘冻干粉0.1%~10%、柠檬酸0.01%~0.6%、果胶0.1%~2%,其余为水。但是该饮料是用牡蛎多糖和其他物质进行复配,很少见到不同提取方式得到的牡蛎多糖进行复配。
发明内容
本发明的目的在于提供一种提取方法简单可行,生产成本低,易于工业化规模生产的牡蛎多糖复配物的制备方法,该制备方法所得具有改善心脏机能、促进血液循环、增强机体免疫力、保护肝脏等效果,可直接被机体吸收,降低对胰腺的负担。
本发明针对上述技术中提到的问题,采取的技术方案为:
牡蛎多糖复配物的制备方法,其具体步骤为:
步骤1:将牡蛎鲜肉绞碎、匀浆,用石油醚脱脂,经低温烘干、粉碎制成脱脂牡蛎粉,多糖的组织细胞多有脂质包围,组织中往往含有较多的蛋白,所以要先进行脱脂处理,该步骤可以除去牡蛎中的脂肪,而脂肪的存在可对产品的风味、色泽等产生不良影响,同时容易造成水解液的浑浊,影响牡蛎多糖的活性;
步骤2:将脱脂牡蛎粉和水按料液比为1:20-30混合均匀,沸水浸提8-12min,过滤,滤渣用同样方法再处理2-3次,合并滤液,调节总滤液pH至5.0-6.0,在8000-10000rpm下离心22-28min,获得的上清液过超滤膜,过膜后的滤液进行喷雾干燥,得到水提取粗多糖,沉淀物烘干,备用,水提取粗多糖具有条件温和、简单等特点,但沉淀物中含有多糖成分,提取率不高,同时超滤可以除去小分子无机盐、低聚糖等杂质,不需要采用其他方法,节约生产成本和能耗;
步骤3:将水提沉淀物中按料液比为1:18-22加入2-5%NaOH溶液,再加入沉淀物重量0.23-0.26%的硼氢化钠和0.014-0.017%的席夫碱,在50-70℃下提取80-100min,然后将调pH至中性,8000-10000rpm下离心22-28min,获得的上清液过超滤膜,过膜后的滤液冻干,得到碱提取粗多糖,水提取后的沉淀物里仍含有多糖物质,牡蛎多糖以和蛋白相结合的糖蛋白形式存在,其共价键遇碱不稳定,碱提法有助于多糖的释放,提高多糖的得率,但多糖分子有可能被碱降解,对多糖的结构和活性有一定的影响,而硼氢化钠和席夫碱的加入一方面能够将醛基还原,保护多糖还原端的单糖不被剥落,避免多糖碱降解现象的发生,使多糖能够保持原有结构,另一方面可以破坏糖蛋白的共价键,加快碱提取速率,进而减小多糖的碱降解;
步骤4:按料液比为1:11-13向水提取粗多糖和碱提取粗多糖加水溶解,调节pH至7.8-8.5,按加酶量为0.4-0.6%加入胰蛋白酶,在50-60℃下反应3-5h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,再调pH至2.0-3.0,按加酶量为0.4-0.6%加入胃蛋白酶,在30-40℃下反应3-5h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,在8000-10000rpm下离心22-28min,获得的上清液加入1/4-1/2体积的Sevage试剂,摇晃20-40min,取上层多糖溶液,重复上述操作3-5次,Sevage脱蛋白得到的糖液中加入3-5倍体积95%乙醇,在2-5℃条件下静置10-15h,离心后取沉淀,干燥脱水2-3次,冻干,即得水提取牡蛎多糖和碱提牡蛎多糖,运用酶解及Sevage试剂联用的方法除蛋白,通过酶解可以除去大部分蛋白,可以减少Sevag法脱蛋白的次数,降低有机试剂的使用,不仅提高了蛋白质的去除率,同时也减少了多糖的损失,该法条件温和,能保存多糖原有的结构与活性;
步骤5:将重量比为1:0.38-0.55的水提取牡蛎多糖和碱提牡蛎多糖进行复配混合,然后制成胶囊剂型,胶囊的规格为0.6-0.7g/粒,每粒胶囊中牡蛎多糖含量为0.22-0.25g,即得牡蛎多糖复配物,该复配物综合水提取牡蛎多糖和碱提取牡蛎多糖的优势,不仅具有较好的DPPH·自由基及羟基自由基清除作用,具有体外清除ABTS+·自由基的能力,对肝损伤具有保护作用,对血管紧张素转化酶(ACE)具有抑制作用,而且具有改善心脏机能、促进血液循环、增强机体免疫力等效果,增效作用明显,综合功效大于单成分多糖,此外该复配物可以直接被机体吸收,降低对胰腺的负担,对糖尿病人十分重要,且胶囊剂型可保持牡蛎多糖的稳定性,崩解后在肠道直接吸收,无需溶解过程,吸收快,生物利用度高,服用方便。
与现有技术相比,本发明的优点在于:1)本发明制备方法简单可行,机械化操作成度高,资源利用率高,提取速率快,能耗低,提取率高,生产成本低,易于工业化规模生产;2)该制备方法对水提取后的沉淀物进行碱提取,提高多糖的得率,避免资源的浪费,提高牡蛎的价值;3)该制备方法采用酶解及Sevage试剂联用的方法除蛋白,通过酶解可以除去大部分蛋白,可以减少Sevag法脱蛋白的次数,降低有机试剂的使用,不仅提高了蛋白质的去除率,同时也减少了多糖的损失,该法条件温和,能保存多糖原有的结构与活性;4)该复配物具有清除DPPH·、羟基和ABTS+·自由基的能力,对肝损伤具有保护作用,具有改善心脏机能、促进血液循环、增强机体免疫力等效果,可直接被机体吸收,降低对胰腺的负担。
具体实施方式
下面通过实施例对本发明方案作进一步说明:
实施例1:
牡蛎多糖复配物的制备方法,其具体步骤为:
步骤1:将牡蛎鲜肉绞碎、匀浆,用石油醚脱脂,经低温烘干、粉碎制成脱脂牡蛎粉;
步骤2:将脱脂牡蛎粉和水按料液比为1:20-30混合均匀,沸水浸提8-12min,过滤,滤渣用同样方法再处理2-3次,合并滤液,调节总滤液pH至5.0-6.0,在8000-10000rpm下离心22-28min,获得的上清液过超滤膜,过膜后的滤液进行喷雾干燥,得到水提取粗多糖,沉淀物烘干,备用;
步骤3:将水提沉淀物中按料液比为1:18-22加入2-5%NaOH溶液,再加入沉淀物重量0.23-0.26%的硼氢化钠和0.014-0.017%的席夫碱,在50-70℃下提取80-100min,然后将调pH至中性,8000-10000rpm下离心22-28min,获得的上清液过超滤膜,过膜后的滤液冻干,得到碱提取粗多糖;
步骤4:按料液比为1:11-13向水提取粗多糖和碱提取粗多糖加水溶解,调节pH至7.8-8.5,按加酶量为0.4-0.6%加入胰蛋白酶,在50-60℃下反应3-5h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,再调pH至2.0-3.0,按加酶量为0.4-0.6%加入胃蛋白酶,在30-40℃下反应3-5h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,在8000-10000rpm下离心22-28min,获得的上清液加入1/4-1/2体积的Sevage试剂,摇晃20-40min,取上层多糖溶液,重复上述操作3-5次,Sevage脱蛋白得到的糖液中加入3-5倍体积95%乙醇,在2-5℃条件下静置10-15h,离心后取沉淀,干燥脱水2-3次,冻干,即得水提取牡蛎多糖和碱提牡蛎多糖;
步骤5:将重量比为1:0.38-0.55的水提取牡蛎多糖和碱提牡蛎多糖进行复配混合,然后制成胶囊剂型,胶囊的规格为0.6-0.7g/粒,每粒胶囊中牡蛎多糖含量为0.22-0.25g,即得牡蛎多糖复配物。
实施例2:
牡蛎多糖复配物的制备方法,其具体步骤为:
步骤1:将牡蛎鲜肉绞碎、匀浆,用石油醚脱脂,经低温烘干、粉碎制成脱脂牡蛎粉;
步骤2:将脱脂牡蛎粉和水按料液比为1:22混合均匀,沸水浸提12min,过滤,滤渣用同样方法再处理3次,合并滤液,调节总滤液pH至6.0,在8000rpm下离心28min,获得的上清液过超滤膜,过膜后的滤液进行喷雾干燥,得到水提取粗多糖,沉淀物烘干,备用;
步骤3:将水提沉淀物中按料液比为1:18加入5%NaOH溶液,再加入沉淀物重量0.23%的硼氢化钠和0.017%的席夫碱,在55℃下提取100min,然后将调pH至中性,8000rpm下离心28min,获得的上清液过超滤膜,过膜后的滤液冻干,得到碱提取粗多糖;
步骤4:按料液比为1: 13向水提取粗多糖和碱提取粗多糖加水溶解,然后再加入粗多糖重量0.033‰的β-氨基乙磺酸和0.051‰的十二烷基苯磺酸钠,调节pH至7.8,按加酶量为0.6%加入胰蛋白酶,在50℃下反应5h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,再调pH至2.0,按加酶量为0.6%加入胃蛋白酶,在33℃下反应5h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,在8000rpm下离心28min,获得的上清液加入1/2体积的Sevage试剂,摇晃20min,取上层多糖溶液,重复上述操作5次,Sevage脱蛋白得到的糖液中加入5倍体积95%乙醇,在5℃条件下静置10h,离心后取沉淀,干燥脱水3次,冻干,即得水提取牡蛎多糖和碱提牡蛎多糖,β-氨基乙磺酸和十二烷基苯磺酸钠的加入能够使多糖结构松散开,暴露出分子内部的酶作用位点,利于蛋白酶水解掉糖蛋白的蛋白质部分,剩下糖链,提高蛋白酶酶解糖蛋白的速度和彻底度,达到意想不到的效果,且其能溶于水排除,无残留,对多糖的活性无不良影响;
步骤5:将重量比为1:0.4的水提取牡蛎多糖和碱提牡蛎多糖进行复配混合,然后制成胶囊剂型,胶囊的规格为0.7g/粒,每粒胶囊中牡蛎多糖含量为0.22g,即得牡蛎多糖复配物。
实施例3:
牡蛎多糖复配物的制备方法,其具体步骤为:
1)将牡蛎鲜肉绞碎、匀浆,用石油醚脱脂,经低温烘干、粉碎制成脱脂牡蛎粉;
2)将脱脂牡蛎粉和水按料液比为1:25混合均匀,沸水浸提10min,过滤,滤渣用同样方法再处理2次,合并滤液,调节总滤液pH至5.5,在9000rpm下离心25min,获得的上清液过超滤膜,过膜后的滤液进行喷雾干燥,得到水提取粗多糖,沉淀物烘干,备用;
3)将水提沉淀物中按料液比为1:20加入3%NaOH溶液,再加入沉淀物重量0.25%的硼氢化钠和0.015%的席夫碱,在60℃下提取90min,然后将调pH至中性,9000rpm下离心25min,获得的上清液过超滤膜,过膜后的滤液冻干,得到碱提取粗多糖;
4)按料液比为1:12向粗多糖加水溶解,调节pH至8.0,按加酶量为0.5%加入胰蛋白酶,在55℃下反应4h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,再调pH至2.5,按加酶量为0.5%加入胃蛋白酶,在37℃下反应4h,反应过程中维持pH,结束后调pH至中性,灭酶,冷却至室温,在9000rpm下离心25min,获得的上清液加入1/3体积的Sevage试剂,摇晃30min,取上层多糖溶液,重复上述操作4次,Sevage脱蛋白得到的糖液中加入4倍体积95%乙醇,在4℃条件下静置13h,离心后取沉淀,干燥脱水2.5次,冻干,即得水提取牡蛎多糖和碱提牡蛎多糖;
5)将重量比为1:0.45的水提取牡蛎多糖和碱提牡蛎多糖进行复配混合,然后制成胶囊剂型,胶囊的规格为0.65g/粒,每粒胶囊中牡蛎多糖含量为0.24g,即得牡蛎多糖复配物。
本发明的操作步骤中的常规操作为本领域技术人员所熟知,在此不进行赘述。
以上所述的实施例对本发明的技术方案进行了详细说明,应理解的是以上所述仅为本发明的具体实施例,并不用于限制本发明,凡在本发明的原则范围内所做的任何修改、补充或类似方式替代等,均应包含在本发明的保护范围之内。
Claims (10)
1.牡蛎多糖复配物的制备方法,包括其特征在于:所述制备方法包括以下步骤:
1)将牡蛎鲜肉绞碎、匀浆,用石油醚脱脂,经低温烘干、粉碎制成脱脂牡蛎粉;
2)将脱脂牡蛎粉和水混合均匀,沸水浸提,过滤,滤渣用同样方法再处理,合并滤液,调节总滤液pH,离心,获得的上清液过超滤膜,过膜后的滤液进行喷雾干燥,得到水提取粗多糖,沉淀物烘干,备用;
3)将水提沉淀物中加入NaOH溶液,再加入硼氢化钠和席夫碱,提取,然后将调pH至中性,离心,获得的上清液过超滤膜,过膜后的滤液冻干,得到碱提取粗多糖;
4)向水提取粗多糖和碱提取粗多糖中分别加水溶解,加入胰蛋白酶反应,结束后调pH至中性,灭酶,冷却至室温,再加入胃蛋白酶反应,结束后调pH至中性,灭酶,冷却至室温,离心,获得的上清液加入Sevage试剂,摇晃,取上层Sevage脱蛋白得到的糖液,加入乙醇,静置,离心后取沉淀,干燥脱水,冻干,即得水提取牡蛎多糖和碱提牡蛎多糖;
5)将水提取牡蛎多糖和碱提牡蛎多糖进行复配混合,然后制成胶囊剂型,即得牡蛎多糖复配物。
2.根据权利要求1所述的牡蛎多糖复配物的制备方法,其特征在于:所述步骤2中脱脂牡蛎粉和水的料液比为1:20-30,浸提8-12min,反复浸提2-3次。
3.根据权利要求1所述的牡蛎多糖复配物的制备方法,其特征在于:所述步骤3中水提沉淀物与NaOH溶液的料液比为1:18-22,NaOH溶液的浓度为2-5%,提取温度为50-70℃、时间为80-100min。
4.根据权利要求1所述的牡蛎多糖复配物的制备方法,其特征在于:所述步骤3中硼氢化钠的添加量为沉淀物重量的0.23-0.26%,席夫碱的添加量为沉淀物重量的0.014-0.017%。
5.根据权利要求1所述的一种牡蛎多糖的提取方法,其特征在于:所述步骤4中水提取粗多糖和碱提取粗多糖与水的料液比均为1:11-13。
6.根据权利要求1所述的一种牡蛎多糖的提取方法,其特征在于:所述步骤4中胰蛋白酶的加酶量为0.4-0.6%,反应pH为7.8-8.5,温度为50-60℃,时间为3-5h。
7.根据权利要求1所述的一种牡蛎多糖的提取方法,其特征在于:所述步骤4中胃蛋白酶的加酶量为0.4-0.6%,反应pH为2.0-3.0,温度为30-40℃,时间为3-5h。
8.根据权利要求1所述的一种牡蛎多糖的提取方法,其特征在于:所述步骤4中Sevage试剂的添加量为酶解液体积的1/4-1/2,摇晃20-40min,重复操作3-5次。
9.根据权利要求1所述的一种牡蛎多糖的提取方法,其特征在于:所述步骤5中水提取牡蛎多糖和碱提牡蛎多糖的重量比为1:0.38-0.55。
10.根据权利要求1所述的一种牡蛎多糖的提取方法,其特征在于:所述步骤5中胶囊的规格为0.6-0.7g/粒,每粒胶囊中牡蛎多糖含量为0.22-0.25g。
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