CN108719981A - 一种微生物酵素钙泡腾片的制备方法 - Google Patents
一种微生物酵素钙泡腾片的制备方法 Download PDFInfo
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Classifications
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Abstract
本发明公开了微生物酵素钙泡腾片的制备方法,属于保健食品领域。本发明利用羊骨为原料,碱性蛋白酶、复合风味蛋白酶进行二级酶解,其富含丰富的氨基酸,再以鸡蛋壳为原料,帮助钙离子螯合,提高谷氨酸的利用率和钙螯合率,能维持钙在小肠内的溶解状态,以干酪素为原料,将胰蛋白酶固定,制备酪蛋白磷酸肽类物质,其能与钙在小肠这种弱碱性环境中形成可溶性复合物,有效防止在中性到偏碱性的小肠环境内不溶性磷酸钙的沉淀,用酿酒酵母、干酪乳杆菌对蓝莓、鲜橙、芹菜进行发酵,制备混合果蔬酵素,促进丰富的蛋白酶、花青素、多酚类营养元素溶出,具有提升机体的抗氧化性、免疫力、降低胆固醇功效,同时通过发酵分解成小分子物质,促进人体吸收。
Description
技术领域
本发明涉及保健食品领域,具体涉及一种微生物酵素钙泡腾片的制备方法。
背景技术
人体中近99%的钙质存在于骨骼和牙齿中,当体液中钙浓度不正常时,血液便会从骨骼中摄取钙,导致骨骼中钙的损失,钙质损失,就会导致骨骼疾病的发生。避免钙质流失,增加钙质吸收便成为促进骨骼健康的重要手段。目前市场上补钙产品种类很多,但由于机体对钙吸收营造的环境比较单一,使钙很难吸收或吸收不完全。一些直接补充钙的产品,由于吸收不好,所补充的钙不易被吸收,反而导致大量的钙流失,极易因缺钙而导致骨骼疾病。人一生中的各阶段都要不断地补钙。儿童缺钙的严重危害:将影响生长发育,严重时会导致儿童佝偻病“X”和“O”型腿等。青少年缺钙的严重危害:生长痛、精力不集中、易过敏、易感冒、偏食烦躁、易疲倦,缺钙会使眼球失去弹性,引起假性近视眼。孕妇及哺乳期妇女缺钙的危害:女性一生都要补钙,年轻时身体发育需要补钙,而孕妇、产妇、哺乳期妇女缺钙将直接影响两代人的健康。老年人缺钙的严重危害:缺钙会使老年人加速衰老,影响晚年幸福,严重者会使老人在病床上度过晚年。现已证明,高血压、动脉硬化、糖尿病、心率紊乱都与缺钙有直接关系。
补钙要坚持科学补钙的原则,目前市场上常用的钙剂有无机钙类及其制剂、有机钙类及其制剂、中药钙类及其它。其中无机钙类(如碳酸钙)是所有药用钙盐中含钙量最高的一种,且价格低廉,是目前应用最广的补钙剂,但此类钙剂可引起嗳气、便秘等不良反应,胃酸缺乏者会影响其吸收;有机钙类(葡萄糖酸钙)的水溶性好,但吸收率低;中药类钙剂(如龙牡壮骨冲剂)的原料大多采用动物骨骼、海洋生物的脊椎、贝壳等,碱性较强,对胃肠刺激大,如果这些生物受到污染,体内可蓄积大量重金属,易造成二次中毒。另外,雌激素替代疗法可预防绝经后妇女骨量减少并降低骨质疏松性骨折的发生,但长期应用存在对生殖系统潜在的致癌风险。因此要科学补钙,药补不如食补,食物摄入的钙来源于牛奶、蔬菜、水果等,但这些食品远远不能满足儿童青少年、孕妇、哺乳期妇女、绝经期妇女及中老年人等特殊人群对钙的需求。
总结来说,现如今市场上的补钙制剂主要存在着以下的缺陷:现有的补钙制剂大多都是从生物体内提取获得的钙元素,但是钙元素的吸收率低,且对人体的肠胃易造成过大的危害,副作用较为明显。
因此,我们有必要重新的设计制备出一种新型的补钙制剂,以此来解决现如今市场山的补钙制剂所存在的一些技术性的缺陷,以此来更好的适应于社会与有关部门的需求。
发明内容
本发明所要解决的技术问题:针对目前现有的补钙制剂大多都是从生物体内提取获得的钙元素,但是钙元素的吸收率低,且对人体的肠胃易造成过大的危害,副作用较为明显的问题,提供一种微生物酵素钙泡腾片的制备方法。
为解决上述技术问题,本发明采用如下所述的技术方案是:
一种微生物酵素钙泡腾片的制备方法,其特征在于,该制备方法包括如下步骤:
(1)取鲜羊骨按质量比2:1加入蒸馏水,于120~125℃保温20~30min,冷却,取出羊骨去除碎肉、骨髓,干燥,粉碎过80目筛,收集过筛颗粒物A,按质量比1:5加入无水乙醇,于25~30℃保温18~24h,旋转蒸发,得旋转蒸发物,按4~5%的添加量加入碱性蛋白酶,酶解,得一级酶解物,取一级酶解物按3~4%的添加量加入复合风味蛋白酶,酶解,灭酶,离心,取上清液,调节pH至7~7.3,得酶解液;
(2)取鸡蛋壳干燥,粉碎过100目筛,收集过筛颗粒物B,再进行气流超微粉碎2~4min,取鸡蛋壳粉按质量比2:3加入盐酸,搅拌混合,过滤,取滤液,调节pH至7~9,取酶解液按质量比2:1加入滤液,于45~55℃搅拌混合50~60min,抽滤,取抽滤液减压浓缩,得浓缩物,取浓缩物按质量比1:8加入无水乙醇,混合,离心,取沉淀干燥,得微生物酵素钙基体物,备用;
(3)取干酪素粉末按质量比2:5加入氢氧化钠溶液,搅拌混合,得混合液A,按质量份数计,取壳聚糖取6~9份壳聚糖、10~12份乙酸、2~3份戊二醛、0.5~0.8份胰蛋白酶、40~50份乙醇溶液混合,于30~35℃搅拌混合,得混合液B,取混合液B按3~5%的添加量加入混合液A中,于40~45℃保持2~3h,调节pH至4.2~4.5,灭酶,离心,取上清液按质量比5:3:10加入氯化钙、无水乙醇搅拌混合,静置5~6h,取沉淀,干燥,得干燥物,取微生物酵素钙基体物按质量比10:3加入干燥物混合,得混合物,备用;
(4)取酿酒酵母菌粉按质量比1:5加入生理盐水混合,按2%的接种量接种至YPD培养基中培养,得酿酒酵母菌活化培养液,取干酪乳杆菌菌粉按质量比1:6加入生理盐水,按3%的接种量接种至MRS培养基中培养,得干酪乳杆菌活化培养液,取酿酒酵母菌活化培养液、干酪乳杆菌活化培养液按质量比1:2混合,得混合菌液;
(5)取蓝莓、鲜橙、芹菜按质量比5:3:2混合,匀浆,取匀浆液灭菌,得灭菌匀浆液,按质量份数计,取0.2~0.5份混合菌液、20~30份灭菌匀浆液、0.2~0.4份酵母膏、0.6~0.9份碳酸铵、1~3份磷酸氢钾、0.02~0.05份硫酸镁,1000份水,发酵,得发酵液,减压浓缩,冷冻干燥,得冷冻干燥物;
(6)按质量份数计,取30~40份步骤(3)备用的混合物、20~30份冷冻干燥物、2~4份柠檬酸、5~7份羟丙甲纤维素、2~3份L-亮氨酸、8~10份乳糖、1~2份β-胡萝卜素、2~4份碳酸氢钠、0.5~0.8份维生素C、100份水混合,压片,制成片剂,得微生物酵素钙泡腾片。
本发明与其他方法相比,有益技术效果是:
(1)本发明利用羊骨为原料,用碱性蛋白酶、复合风味蛋白酶进行二级酶解,得到酶解胶原多肽液,其富含丰富的氨基酸类物质,再以鸡蛋壳为原料,超微粉碎,较小的粒子可以填充在一些较大的颗粒空隙中,有助于钙离子充分螯合,从而提高谷氨酸的利用率和螯合钙的螯合率,粉体比表面积变大,粉体容易吸附在谷氨酸表面,有利于螯合物的形成酸提其中的钙元素,以钙离子为中心离子,胶原多肽氨基酸基团中的羧基的氧原子和氨基的氮原子为配位体,多肽与钙形成的螯合物由于其独特的鳌合体制和转运机制,能维持钙在小肠内的溶解状态,增加小肠对钙的吸收及其在体内的蓄积,使肽钙螯合物比氨基酸钙鳌合物更易于被吸收且具有更快的吸收速度;
(2)本发明以干酪素为原料,将胰蛋白酶进行固定化,制备一种酪蛋白磷酸肽类物质,因对二价金属的亲和性,能与钙在小肠这种弱碱性环境中形成可溶性复合物,这种结合能有效防止在中性到偏碱性的小肠环境内不溶性磷酸钙的沉淀,增加可溶性钙的浓度,从而促进肠内钙的吸收,其可促进小肠下部不饱和钙的被动扩散吸收,不受年龄的影响,一方面对钙进行了保护,提高了钙的利用吸收率,另一方面不会产生任何毒副作用,对人体安全;
(3)本发明利用蓝莓、鲜橙、芹菜为原料,利用酿酒酵母、干酪乳杆菌进行发酵,制备混合果蔬酵素,通过发酵,促进其中的营养元素溶出,配合钙制剂进行营养的补充,满足人体的多元化营养需求,其含有丰富的蛋白酶、花青素、多酚类物质,具有提升机体的抗氧化性、免疫力、降低胆固醇的功效,同时通过发酵分解成小分子物质,促进人体吸收。
具体实施方式
复合风味蛋白酶:购于忆诺联创生物科技发展(北京)有限公司。
干酪素粉末:购于成都市科龙化工试剂厂。
酿酒酵母菌粉:购于湖北省安琪酵母股份有限公司;干酪乳杆菌粉:购于内蒙古普泽生物制品有限责任公司。
一种微生物酵素钙泡腾片的制备方法,包括如下步骤:
(1)取鲜羊骨按质量比2:1加入蒸馏水,于120~125℃保持20~30min,冷却,取出羊骨去除碎肉、骨髓,于100~110℃干燥5~6h,粉碎过80目筛,收集过筛颗粒物A,按质量比1:5加入无水乙醇,于25~30℃保持18~24h,旋转蒸发,得旋转蒸发物,按4~5%的添加量加入碱性蛋白酶,于45~50℃酶解6~7h,得一级酶解物,取一级酶解物按3~4%的添加量加入复合风味蛋白酶,于50~55℃酶解4~5h,90~95℃灭酶15~20min,离心,取上清液,调节pH至7~7.3,得酶解液;
(2)取鸡蛋壳干燥,粉碎过100目筛,收集过筛颗粒物B,再进行气流超微粉碎2~4min,取鸡蛋壳粉按质量比2:3加入浓度为1mol/L的盐酸,搅拌混合10~12h,过滤,取滤液,调节pH至7~9,取酶解液按质量比2:1加入滤液,于45~55℃搅拌混合50~60min,抽滤,取抽滤液减压浓缩,得浓缩物,取浓缩物按质量比1:8加入无水乙醇,混合2~3h,离心,取沉淀干燥,得微生物酵素钙基体物,备用;
(3)取干酪素粉末按质量比2:5加入质量分数为20%的氢氧化钠溶液,搅拌混合20~30min,得混合液A,按质量份数计,取壳聚糖取6~9份壳聚糖、10~12份乙酸、2~3份戊二醛、0.5~0.8份胰蛋白酶、40~50份质量分数为70%的乙醇溶液混合,于30~35℃搅拌混合3~6h,得混合液B,取混合液B按3~5%的添加量加入混合液A中,于40~45℃保持2~3h,调节pH至4.2~4.5,灭酶,离心,取上清液按质量比5:3:10加入氯化钙、无水乙醇搅拌混合,静置5~6h,取沉淀,干燥,得干燥物,取微生物酵素钙基体物按质量比10:3加入干燥物混合,得混合物,备用;
(4)取酿酒酵母菌粉按质量比1:5加入生理盐水混合,按2%的接种量接种至YPD培养基中,于28~33℃培养3~5天,得酿酒酵母菌活化培养液,取干酪乳杆菌菌粉按质量比1:6加入生理盐水,按3%的接种量接种至MRS培养基中,于30~35℃培养2~4天,得干酪乳杆菌活化培养液,取酿酒酵母菌活化培养液、干酪乳杆菌活化培养液按质量比1:2混合,得混合菌液;
(5)取蓝莓、鲜橙、芹菜按质量比5:3:2混合,匀浆,取匀浆液灭菌,得灭菌匀浆液,按质量份数计,取0.2~0.5份混合菌液、20~30份灭菌匀浆液、0.2~0.4份酵母膏、0.6~0.9份碳酸铵、1~3份磷酸氢钾、0.02~0.05份硫酸镁,1000份水,于28~35℃发酵3~5天,得发酵液,减压浓缩,冷冻干燥,得冷冻干燥物;
(6)按质量份数计,取30~40份步骤(3)备用的混合物、20~30份冷冻干燥物、2~4份柠檬酸、5~7份羟丙甲纤维素、2~3份L-亮氨酸、8~10份乳糖、1~2份β-胡萝卜素、2~4份碳酸氢钠、0.5~0.8份维生素C、100份水混合,压片,制成片剂,得微生物酵素钙泡腾片。
实施例1
复合风味蛋白酶:购于忆诺联创生物科技发展(北京)有限公司。
干酪素粉末:购于成都市科龙化工试剂厂。
酿酒酵母菌粉:购于湖北省安琪酵母股份有限公司;干酪乳杆菌粉:购于内蒙古普泽生物制品有限责任公司。
一种微生物酵素钙泡腾片的制备方法,包括如下步骤:
(1)取鲜羊骨按质量比2:1加入蒸馏水,于120℃保持20min,冷却,取出羊骨去除碎肉、骨髓,于100℃干燥5h,粉碎过80目筛,收集过筛颗粒物A,按质量比1:5加入无水乙醇,于25℃保持18h,旋转蒸发,得旋转蒸发物,按4%的添加量加入碱性蛋白酶,于45℃酶解6h,得一级酶解物,取一级酶解物按3%的添加量加入复合风味蛋白酶,于50℃酶解4h,90℃灭酶15min,离心,取上清液,调节pH至7,得酶解液;
(2)取鸡蛋壳干燥,粉碎过100目筛,收集过筛颗粒物B,再进行气流超微粉碎2min,取鸡蛋壳粉按质量比2:3加入浓度为1mol/L的盐酸,搅拌混合10h,过滤,取滤液,调节pH至7,取酶解液按质量比2:1加入滤液,于45℃搅拌混合50min,抽滤,取抽滤液减压浓缩,得浓缩物,取浓缩物按质量比1:8加入无水乙醇,混合2h,离心,取沉淀干燥,得微生物酵素钙基体物,备用;
(3)取干酪素粉末按质量比2:5加入质量分数为20%的氢氧化钠溶液,搅拌混合20min,得混合液A,按质量份数计,取壳聚糖取6份壳聚糖、10份乙酸、2份戊二醛、0.5份胰蛋白酶、40份质量分数为70%的乙醇溶液混合,于30℃搅拌混合3h,得混合液B,取混合液B按3~5%的添加量加入混合液A中,于40℃保持2~3h,调节pH至4.2,灭酶,离心,取上清液按质量比5:3:10加入氯化钙、无水乙醇搅拌混合,静置5h,取沉淀,干燥,得干燥物,取微生物酵素钙基体物按质量比10:3加入干燥物混合,得混合物,备用;
(4)取酿酒酵母菌粉按质量比1:5加入生理盐水混合,按2%的接种量接种至YPD培养基中,于28℃培养3天,得酿酒酵母菌活化培养液,取干酪乳杆菌菌粉按质量比1:6加入生理盐水,按3%的接种量接种至MRS培养基中,于30℃培养2天,得干酪乳杆菌活化培养液,取酿酒酵母菌活化培养液、干酪乳杆菌活化培养液按质量比1:2混合,得混合菌液;
(5)取蓝莓、鲜橙、芹菜按质量比5:3:2混合,匀浆,取匀浆液灭菌,得灭菌匀浆液,按质量份数计,取0.2份混合菌液、20份灭菌匀浆液、0.2份酵母膏、0.6份碳酸铵、1份磷酸氢钾、0.02份硫酸镁,1000份水,于28℃发酵3天,得发酵液,减压浓缩,冷冻干燥,得冷冻干燥物;
(6)按质量份数计,取30份步骤(3)备用的混合物、20份冷冻干燥物、2份柠檬酸、5份羟丙甲纤维素、2份L-亮氨酸、8份乳糖、1份β-胡萝卜素、2份碳酸氢钠、0.5份维生素C、100份水混合,压片,制成片剂,得微生物酵素钙泡腾片。
实施例2
复合风味蛋白酶:购于忆诺联创生物科技发展(北京)有限公司。
干酪素粉末:购于成都市科龙化工试剂厂。
酿酒酵母菌粉:购于湖北省安琪酵母股份有限公司;干酪乳杆菌粉:购于内蒙古普泽生物制品有限责任公司。
一种微生物酵素钙泡腾片的制备方法,包括如下步骤:
(1)取鲜羊骨按质量比2:1加入蒸馏水,于123℃保持25min,冷却,取出羊骨去除碎肉、骨髓,于105℃干燥5.5h,粉碎过80目筛,收集过筛颗粒物A,按质量比1:5加入无水乙醇,于28℃保持20h,旋转蒸发,得旋转蒸发物,按4.5%的添加量加入碱性蛋白酶,于48℃酶解6.5h,得一级酶解物,取一级酶解物按3.5%的添加量加入复合风味蛋白酶,于53℃酶解4.5h,93℃灭酶18min,离心,取上清液,调节pH至7.2,得酶解液;
(2)取鸡蛋壳干燥,粉碎过100目筛,收集过筛颗粒物B,再进行气流超微粉碎3min,取鸡蛋壳粉按质量比2:3加入浓度为1mol/L的盐酸,搅拌混合11h,过滤,取滤液,调节pH至8,取酶解液按质量比2:1加入滤液,于50℃搅拌混合55min,抽滤,取抽滤液减压浓缩,得浓缩物,取浓缩物按质量比1:8加入无水乙醇,混合2.5h,离心,取沉淀干燥,得微生物酵素钙基体物,备用;
(3)取干酪素粉末按质量比2:5加入质量分数为20%的氢氧化钠溶液,搅拌混合25min,得混合液A,按质量份数计,取壳聚糖取7份壳聚糖、11份乙酸、2.5份戊二醛、0.7份胰蛋白酶、45份质量分数为70%的乙醇溶液混合,于33℃搅拌混合5h,得混合液B,取混合液B按4%的添加量加入混合液A中,于43℃保持2.5h,调节pH至4.4,灭酶,离心,取上清液按质量比5:3:10加入氯化钙、无水乙醇搅拌混合,静置5.5h,取沉淀,干燥,得干燥物,取微生物酵素钙基体物按质量比10:3加入干燥物混合,得混合物,备用;
(4)取酿酒酵母菌粉按质量比1:5加入生理盐水混合,按2%的接种量接种至YPD培养基中,于30℃培养4天,得酿酒酵母菌活化培养液,取干酪乳杆菌菌粉按质量比1:6加入生理盐水,按3%的接种量接种至MRS培养基中,于33℃培养3天,得干酪乳杆菌活化培养液,取酿酒酵母菌活化培养液、干酪乳杆菌活化培养液按质量比1:2混合,得混合菌液;
(5)取蓝莓、鲜橙、芹菜按质量比5:3:2混合,匀浆,取匀浆液灭菌,得灭菌匀浆液,按质量份数计,取0.4份混合菌液、25份灭菌匀浆液、0.3份酵母膏、0.7份碳酸铵、2份磷酸氢钾、0.03份硫酸镁,1000份水,于30℃发酵4天,得发酵液,减压浓缩,冷冻干燥,得冷冻干燥物;
(6)按质量份数计,取35份步骤(3)备用的混合物、25份冷冻干燥物、3份柠檬酸、6份羟丙甲纤维素、2.5份L-亮氨酸、9份乳糖、1.5份β-胡萝卜素、3份碳酸氢钠、0.7份维生素C、100份水混合,压片,制成片剂,得微生物酵素钙泡腾片。
实施例3
复合风味蛋白酶:购于忆诺联创生物科技发展(北京)有限公司。
干酪素粉末:购于成都市科龙化工试剂厂。
酿酒酵母菌粉:购于湖北省安琪酵母股份有限公司;干酪乳杆菌粉:购于内蒙古普泽生物制品有限责任公司。
一种微生物酵素钙泡腾片的制备方法,包括如下步骤:
(1)取鲜羊骨按质量比2:1加入蒸馏水,于125℃保持30min,冷却,取出羊骨去除碎肉、骨髓,于110℃干燥6h,粉碎过80目筛,收集过筛颗粒物A,按质量比1:5加入无水乙醇,于30℃保持24h,旋转蒸发,得旋转蒸发物,按5%的添加量加入碱性蛋白酶,于50℃酶解7h,得一级酶解物,取一级酶解物按4%的添加量加入复合风味蛋白酶,于50~55℃酶解5h,95℃灭酶20min,离心,取上清液,调节pH至7.3,得酶解液;
(2)取鸡蛋壳干燥,粉碎过100目筛,收集过筛颗粒物B,再进行气流超微粉碎4min,取鸡蛋壳粉按质量比2:3加入浓度为1mol/L的盐酸,搅拌混合12h,过滤,取滤液,调节pH至9,取酶解液按质量比2:1加入滤液,于55℃搅拌混合60min,抽滤,取抽滤液减压浓缩,得浓缩物,取浓缩物按质量比1:8加入无水乙醇,混合3h,离心,取沉淀干燥,得微生物酵素钙基体物,备用;
(3)取干酪素粉末按质量比2:5加入质量分数为20%的氢氧化钠溶液,搅拌混合20~30min,得混合液A,按质量份数计,取壳聚糖取9份壳聚糖、12份乙酸、3份戊二醛、0.8份胰蛋白酶、50份质量分数为70%的乙醇溶液混合,于35℃搅拌混合6h,得混合液B,取混合液B按5%的添加量加入混合液A中,于45℃保持3h,调节pH至4.5,灭酶,离心,取上清液按质量比5:3:10加入氯化钙、无水乙醇搅拌混合,静置6h,取沉淀,干燥,得干燥物,取微生物酵素钙基体物按质量比10:3加入干燥物混合,得混合物,备用;
(4)取酿酒酵母菌粉按质量比1:5加入生理盐水混合,按2%的接种量接种至YPD培养基中,于33℃培养5天,得酿酒酵母菌活化培养液,取干酪乳杆菌菌粉按质量比1:6加入生理盐水,按3%的接种量接种至MRS培养基中,于35℃培养4天,得干酪乳杆菌活化培养液,取酿酒酵母菌活化培养液、干酪乳杆菌活化培养液按质量比1:2混合,得混合菌液;
(5)取蓝莓、鲜橙、芹菜按质量比5:3:2混合,匀浆,取匀浆液灭菌,得灭菌匀浆液,按质量份数计,取0.5份混合菌液、30份灭菌匀浆液、0.4份酵母膏、0.9份碳酸铵、3份磷酸氢钾、0.05份硫酸镁,1000份水,于35℃发酵5天,得发酵液,减压浓缩,冷冻干燥,得冷冻干燥物;
(6)按质量份数计,取40份步骤(3)备用的混合物、30份冷冻干燥物、4份柠檬酸、7份羟丙甲纤维素、3份L-亮氨酸、10份乳糖、2份β-胡萝卜素、4份碳酸氢钠、0.8份维生素C、100份水混合,压片,制成片剂,得微生物酵素钙泡腾片。
对比例:北京某生物科技公司生产的微生物酵素钙泡腾片。
选取4周龄的清洁大鼠30只,体重80~90g,随机分成三组,每组各10只;空白组大鼠为对照组,喂基础饲料,对比例大鼠喂添加有北京某生物科技公司生产的微生物酵素钙泡腾片的基础饲料,实施例1~3大鼠喂添加有本发明的微生物酵素钙泡腾片的基础饲料;同时用EDTA法测定基础饲料、添加有北京某生物科技公司生产的微生物酵素钙泡腾片的基础饲料、添加有本发明的微生物酵素钙泡腾片的基础饲料的钙含量,结果分别为0.91mg/ g、8.02mg/g、8.33mg/g。
喂养2周后,进行为期3天的代谢试验,准确记录饲料摄入量及粪、尿排出量,并分别混匀留样,用EDTA法测定试验大鼠排泄粪便的钙含量,而钙存留率=(饲
料钙摄入量-排出钙量)/饲料钙摄入量×100%,各项试验数据平均值如下表1所示:
表1:
由上表可知,实施例1~3的钙吸收率远远超过了对比例,因为动物体内的钙留存率往往反映了动物对钙的代谢吸收率,因此可知,大鼠对本发明的微生物酵素钙泡腾片中钙的吸收率要远大于它们对普通补钙剂中钙的吸收率,钙元素的吸收率高。
本发明的最大耐受量试验
按照国家新药报批要求,分别用小鼠和大鼠两种动物口服给药途径设计了大鼠和小鼠对本发明的最大耐受量试验,测定其最大用药量。结果表明,本发明对大鼠一日1次灌胃的剂量大于15g/kg,对大鼠一日3次灌胃的剂量大于45g/kg,对小鼠一日1次灌胃的剂量大于50g/kg,对小鼠一日3次灌胃的剂量大于150g/kg,均未见小鼠、大鼠有异常反应。生活状态依然良好。这表明,本发明实际无毒,安全可靠。
Claims (1)
1.一种微生物酵素钙泡腾片的制备方法,其特征在于,该制备方法包括如下步骤:
(1)取鲜羊骨按质量比2:1加入蒸馏水,于120~125℃保温20~30min,冷却,取出羊骨去除碎肉、骨髓,干燥,粉碎过80目筛,收集过筛颗粒物A,按质量比1:5加入无水乙醇,于25~30℃保温18~24h,旋转蒸发,得旋转蒸发物,按4~5%的添加量加入碱性蛋白酶,酶解,得一级酶解物,取一级酶解物按3~4%的添加量加入复合风味蛋白酶,酶解,灭酶,离心,取上清液,调节pH至7~7.3,得酶解液;
(2)取鸡蛋壳干燥,粉碎过100目筛,收集过筛颗粒物B,再进行气流超微粉碎2~4min,取鸡蛋壳粉按质量比2:3加入盐酸,搅拌混合,过滤,取滤液,调节pH至7~9,取酶解液按质量比2:1加入滤液,于45~55℃搅拌混合50~60min,抽滤,取抽滤液减压浓缩,得浓缩物,取浓缩物按质量比1:8加入无水乙醇,混合,离心,取沉淀干燥,得微生物酵素钙基体物,备用;
(3)取干酪素粉末按质量比2:5加入氢氧化钠溶液,搅拌混合,得混合液A,按质量份数计,取壳聚糖取6~9份壳聚糖、10~12份乙酸、2~3份戊二醛、0.5~0.8份胰蛋白酶、40~50份乙醇溶液混合,于30~35℃搅拌混合,得混合液B,取混合液B按3~5%的添加量加入混合液A中,于40~45℃保持2~3h,调节pH至4.2~4.5,灭酶,离心,取上清液按质量比5:3:10加入氯化钙、无水乙醇搅拌混合,静置5~6h,取沉淀,干燥,得干燥物,取微生物酵素钙基体物按质量比10:3加入干燥物混合,得混合物,备用;
(4)取酿酒酵母菌粉按质量比1:5加入生理盐水混合,按2%的接种量接种至YPD培养基中培养,得酿酒酵母菌活化培养液,取干酪乳杆菌菌粉按质量比1:6加入生理盐水,按3%的接种量接种至MRS培养基中培养,得干酪乳杆菌活化培养液,取酿酒酵母菌活化培养液、干酪乳杆菌活化培养液按质量比1:2混合,得混合菌液;
(5)取蓝莓、鲜橙、芹菜按质量比5:3:2混合,匀浆,取匀浆液灭菌,得灭菌匀浆液,按质量份数计,取0.2~0.5份混合菌液、20~30份灭菌匀浆液、0.2~0.4份酵母膏、0.6~0.9份碳酸铵、1~3份磷酸氢钾、0.02~0.05份硫酸镁,1000份水,发酵,得发酵液,减压浓缩,冷冻干燥,得冷冻干燥物;
(6)按质量份数计,取30~40份步骤(3)备用的混合物、20~30份冷冻干燥物、2~4份柠檬酸、5~7份羟丙甲纤维素、2~3份L-亮氨酸、8~10份乳糖、1~2份β-胡萝卜素、2~4份碳酸氢钠、0.5~0.8份维生素C、100份水混合,压片,制成片剂,得微生物酵素钙泡腾片。
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