CN111034969A - 一种蜂产品胶囊的制备方法 - Google Patents
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Abstract
本发明公开了一种蜂产品胶囊的制备方法,属于蜂产品加工技术领域。本发明所述方法将新鲜的刚从蜂巢取出的蜂蛹立即放入‑80℃速冻;将胶体磨先与冰水循环冷却连接好,然后把速冻蜂蛹过胶体磨得到蜂蛹匀浆液;将蜂蛹匀浆液、蜂王浆、蜂蜜按质量比5:3:2的比例加入胶体磨中均质,得到蜂合剂;在蜂合剂加入辅料麦芽糊精和硬脂酸镁放入真空冷冻干燥机中冻干,将冻干粉过筛、灭菌、装入胶囊壳得到蜂产品胶囊。本发明所述方法制备简单但有效延长了蜂蛹的保质期,保持了蜂产品天然的特性,能较好的发挥其功能活性;本发明蜂产品胶囊中同时加入了蜂蛹匀浆液、蜂王浆、蜂蜜,长期服用可以起到对肾脏的保护效果。
Description
技术领域
本发明涉及一种蜂产品胶囊的制备方法,属于蜂产品加工技术领域。
背景技术
蜂王浆、蜂蜜、蜂蛹等蜂产品被认为是天然抗氧化剂的潜在来源,其能够抵消多种氧化应激引起的疾病。蜂王浆是由工蜂的下咽和下颌腺产生的分泌物。具有血管舒张和降血压活性,诱导血清胆固醇水平降低,抗菌,抗过敏,抗炎症,免疫调节和抗氧化特性。蜂蜜是工蜂经过唾液腺内淀粉酶作用而酿成的一种天然的甜味食品,具有抗炎、抗菌等作用。蜂蛹是蜂王产的卵,孵化成的幼虫,一般包括蜂王、工蜂、雄峰的幼虫及蛹。蜂幼虫身体柔软,呈黄色有褶皱。
蜂蛹营养丰富蛋白质中含有18种氨基酸,人体必需的8种氨基酸种类齐全经研究发现蜂蛹具有抗氧化、抗疲劳、抗衰老、抗炎症、抑制皮炎等功效。蜂蛹的营养丰富,但是在离开蜂房后很容易受到微生物污染变质,尤其是蜂蛹中的络氨酸酶在空气中氧的作用下,其活性增强,络氨酸转化为二羟苯丙氨酸,继而转化为黑色素使蛹体变黑。蜂蛹在离开蜂房2小时即可能发生变质而失去营养价值,而蜂蛹在破损时更是会在数分钟内变黑,因此如何做好蜂蛹的保鲜和加工是现在急需解决的技术问题。
虽然蜂王浆、蜂蜜、蜂蛹营养价值很高,但是一般都是单独食用,食用起来非常不方便,且效果不明显,尤其针对肾损伤的细胞修复并没有很好的效果。
发明内容
本发明的目的在于提供一种蜂产品胶囊的制备方法,具体包括以下步骤:
(1)将新鲜的刚从蜂巢取出的蜂蛹立即放入-80℃速冻;
(2)将胶体磨先与冰水循环冷却连接好,然后把速冻蜂蛹过胶体磨得到蜂蛹匀浆液;
(3)将蜂蛹匀浆液、蜂王浆、蜂蜜按质量比5:3:2的比例加入胶体磨中均质,得到蜂合剂;
(4)在蜂合剂加入辅料麦芽糊精和硬脂酸镁放入真空冷冻干燥机中冻干,将冻干粉过筛、灭菌、装入胶囊壳得到蜂产品胶囊。
优选的,本发明步骤(4)中麦芽糊精的加入量为蜂合剂质量的50%,硬脂酸镁的加入量为蜂合剂质量的1%。
优选的,本发明步骤(4)冻干的条件为:温度为-80℃,直至完全冻干成粉末(10-48h),降温速率为快速降温,1h降至目标温度。
本发明所述胶囊中含蜂合剂的剂量为0.089g/kg,一粒药是0.5g,一天8~10粒
本发明的有益效果:
(1)本发明所述方法将蜂蛹进行速冻处理、全程低温,大大降低了蜂蛹在加工时的变质风险,本发明所述方法制得的蜂合剂胶囊,携带方便,易于服用,活性成分含量高,蜂蛹、蜂王浆的营养保留的更充分,延长了常温下的保质期。
(2)目前蜂蛹产品大多需要添加大量的化工合成的防腐剂,才能保持其活性,本发通过添加蜂王浆和蜂蜜来增加防腐性能;仅仅需要加入少量的辅料来确保冻干效果,制备简单但有效延长了蜂蛹的保质期(一般时间常温为12-24h,本发明的蜂产品保质期为18-24个月),保持了蜂产品天然的特性,能较好的发挥其功能活性。
(3)本发明蜂产品胶囊中同时加入了蜂蛹匀浆液、蜂王浆、蜂蜜,具有抗氧化活性能显著增加肾组织中谷胱甘肽的含量,提高超氧化物歧化酶活力,降低血肌酐、血尿素氮,长期服用可以起到对肾脏的保护效果,且比单一使用蜂蛹匀浆液、蜂王浆、蜂蜜效果要好。此外该蜂产品胶囊具有非常强的抗氧化活性,是非常好的机体或细胞抗氧化剂。
附图说明
图1为小鼠体重变化及肾脏系数;
图2为小鼠肾组织抗氧化指标;
图3为小鼠血浆中肌酐(SCr)和尿素氮(BUN)含量柱状图;
图4为小鼠尿中肌酐、尿素氮、尿蛋白含量柱状图;
图5为小鼠肾组织切片图,图中:肾小球(g),非正常肾小球(ag),近端小管(p),刷状缘(黑箭头),刷状缘丢失(白箭头),细胞质缺失造成细胞空泡化(箭头),管腔内碎屑集聚(星形)。
具体实施方式
下面结合具体实施例本发明作进一步的详细说明,但本发明的保护范围并不限于所述内容。
实施例1
一种蜂产品胶囊的制备方法,具体包括以下步骤:
(1)将新鲜的刚从蜂巢取出的蜂蛹立即放入-80℃速冻;
(2)将胶体磨先与冰水循环冷却连接好,然后把速冻蜂蛹过胶体磨得到蜂蛹匀浆液;
(3)将蜂蛹匀浆液、蜂王浆、蜂蜜按质量比5:3:2的比例加入胶体磨中均质,得到蜂合剂;
(4)在蜂合剂加入辅料麦芽糊精和硬脂酸镁放入温度为-50℃的真空冷冻干燥机中冻干,将冻干粉过筛、灭菌、装入胶囊壳得到蜂产品胶囊;其中麦芽糊精的加入量为蜂合剂质量的50%,硬脂酸镁的加入量为蜂合剂质量的1%。
本发明制备得到的蜂产品胶囊同时加入了蜂蛹匀浆液、蜂王浆、蜂蜜,对肾脏有很好的保护效果,且比单一使用蜂蛹匀浆液、蜂王浆、蜂蜜效果要好,下面通过动物实验进一步验证:
1、制备动物模型
将42只体重相近的八周龄小鼠随机分为五组,每组6只。空白对照组(灌胃、注射生理盐水);模型组(灌胃生理盐水+注射顺铂);阳性药物组即氨磷汀组(灌胃生理盐水+注射顺铂,并在注射顺铂前20min注射氨磷汀400mg/m2);合剂组:配方为将蜂蛹冻干粉加等体积水稀释后,按蜂蛹液100g+蜂王浆60g+蜂蜜40g胶体磨混匀得合剂(灌胃合剂+注射顺铂);蜂蛹组(灌胃蜂蛹+注射顺铂);蜂蜜组(灌胃蜂蜜+注射顺铂);蜂王浆组(灌胃蜂王浆+注射顺铂);具体操作为前7天空白对照组、模型组、氨磷汀组灌胃生理盐水每只0.2ml/d;合剂组每只灌胃合剂0.2ml/d;蜂蛹组、蜂蜜组、蜂王浆组每只灌胃相应蜂产品0.2ml/d;第八天开始每只注射顺铂20mg/m2间隔时间为1天(共4次),期间继续灌胃同剂量生理盐水、合剂、蜂蛹匀浆液(共7天),监控摄食(每天灌胃后将颗粒饲料补至充足且相近的量,次日早观察剩余情况);于实验开始前一天、第8天、第15天记录灌胃前体重。
上述实验中采用的顺铂及氨磷汀剂量均为表面积剂量,具体小鼠体表面积计算公式为A=K·W2/3(A为表面积,单位m2;K为体型系数,新法中小鼠K值为0.0899;W为体重,单位kg。)本实施例用的剂量为一般剂量20mg/m2,按说明书所述“腹腔注射给药器官内的药物浓度是静脉注射的2.5-8倍”,所以顺铂腹腔注射反复间隔给药的方式,可以提供一种低药量,短时间的顺铂致小鼠脏器损伤模型。氨磷汀(Amifostine)是第一个被国际权威机构认可的广谱细胞保护剂,经大量研究表明氨磷汀对顺铂所致的肾毒性具有保护作用,一般需在化疗前使用,价格昂贵。氨磷汀一方面治疗机制就是在体内结合碱性磷酸酯酶代谢为含巯基的WR-1065,降低谷胱甘肽(GSH)的消耗达到保护效果。
2、实验动物的生物标本的获取
在实验第15天采用尿液收集装置,给予每只小鼠1ml水负荷,收集即时尿。
在实验第15天取尿后将小鼠麻醉后取血,断颈处死小鼠,取双肾称重,左侧的肾加入9倍生理盐水(1g:9ml)匀浆,在转速为3500r/min的条件下4℃离心,获得肾组织匀浆上清液,使用试剂盒测定组织匀浆上清液中的SOD(超氧化物歧化酶)、GSH-PX(谷胱甘肽过氧化物酶)、GSH(谷胱甘肽)、MDA(丙二醛)及血、尿中的Cr(肌酐)、BUN(尿素氮)及Bradford法(考马斯亮蓝法)测尿中蛋白。
右侧的肾用中性甲醛固定,固定于2.5%戊二醛溶液中;脱水,透明后,石蜡包埋切片,脱蜡复水,苏木精,依红(HE)染色,光学显微镜下观察肝脏和肾脏的组织形态学改变。
3、实验结果
①小鼠体重及肾脏指数
如图1所示,图1中图A整个实验过程中小鼠体重变化;分别为第0天、第8天、第15天各组小鼠体重;图B为注射顺铂后各组小鼠体重变化;结果使用单因素方差分析,星号表示与空白组比较(图中*表示P<0.05;**表示P<0.01;***表示P<0.001。)图C小鼠肾脏系数,双侧肾质量与体重的比表示为g/100g。从图中可以看出在第8天给予顺铂注射后与正常组相比其他各组小鼠体重明显下降(图A,B),且肾脏器系数升高(图C)表明在给予顺铂注射后的小鼠发生实质性肾损伤,监控摄食发现除合剂组外各造模小鼠食量下降明显,蜂蜜组体重降低最严重,蜂蜜及蜂王浆组肾脏系数明显升高;和其他组相比,合剂组对小鼠体重变化和肾脏系数的影响与氨磷汀组相近,并优于其他组,说明合剂组对顺铂致肾损伤肾脏具有明显的保护效果。
②小鼠抗氧化活性指标
小鼠肾组织抗氧化指标如图2所示,图2A为肾脏中谷胱甘肽含量;图2B为肾脏中谷胱甘肽过氧化物酶活性;图2C为肾脏中总SOD活性;图2D为肾脏中MDA含量。星号表示与空白组比较;井号表示与模型组比较(图中*、#表示P<0.05;**、##表示P<0.01;***、###表示P<0.001)。由图可以看出,合剂组中谷胱甘肽(GSH)的含量显著高于其他组,蜂蜜组的谷胱甘肽过氧化物酶(GSH-PX)、超氧化物歧化酶活性的活性也显著高于其他组,氨磷汀组具有较高的超氧化物歧化酶活性,而各组小鼠肾组织中的丙二醛(MDA)含量无显著差异,但蜂蜜组脂质过氧化物的含量较高,说明其发生氧化损伤,蜂王浆组则相对较低,和其他组相比,合剂组能够显著增加肾脏中的谷胱甘肽含量、增加总SOD的活性,并且谷胱甘肽过氧化物酶活性和丙二醛(MDA)含量与氨磷汀组相当。说明合剂组通过提高肾脏抗氧化活性,对顺铂致肾损伤肾脏具有明显的保护效果。
③小鼠生化指标
如图3所示,图3A为小鼠血浆中肌酐(SCr)的含量;图3B为小鼠血浆中的尿素氮(BUN)含量;星号表示与空白组比较;井号表示与模型组比较(图中*、#表示P<0.05;**、##表示P<0.01;***、###表示P<0.001);从图3可以看出模型组和蜂蛹小鼠血肌酐显著高于空白组(p<0.05),说明单用蜂蛹时,不能很好降低血肌酐。模型组和氨磷汀组血尿素氮水平显著高于空白组(p<0.001)各蜂产品组血尿素氮均值低于氨磷汀组,显示出较好的保护效果。模型组和蜂王浆组小鼠血肌酐显著高于空白(p<0.05;p<0.01),合剂组最接近于空白组尿肌酐水平,如图4所示(图4A为小鼠尿中肌酐(Cr)的含量;图4B为小鼠尿中尿素氮(UN);图C小鼠尿中尿蛋白。星号表示与空白组比较;井号表示与模型组比较(图中*、#表示P<0.05;**、##表示P<0.01;***、###表示P<0.001)。和其他组相比,合剂组能够显著降低血肌酐、血尿素氮、尿中肌酐和尿素氮的水平,且效果好于氨磷汀组,说明合剂组对顺铂致肾损伤肾脏具有明显的保护效果。
④肾组织HE染色结果
正常组小鼠肾具有典型的肾单位结构,如具有刷状缘的正常肾小管,完整的肾小球;而模型组则显示出实质性的肾损伤,如刷状缘丢失;肾小球萎缩;肾小管内碎屑堆积;以及细胞质丢失导致空泡化,氨磷汀组及合剂组显示出较好的治疗效果,如典型的肾小球结构;管腔内未出现碎屑堆积,且相对于氨磷汀组,合剂组显示出较轻的细胞质缺失和刷状缘丢失情况,蜂蛹组具有典型的肾小球结构,但细胞碎屑堆积,细胞质缺失更为严重(如图5所示)。和其他组相比,合剂组能显著改善肾组织的损伤、稳定典型的肾小球结构,且治疗效果好于氨磷汀组。说明合剂组对顺铂致肾损伤肾脏具有明显的保护效果。
综上所述,单一蜂产品,存在相应的缺陷,如蜂蜜虽然能显著提高SOD、GSH-PX的活性,但却存在脂质过氧化物升高,肾脏系数升高的风险;蜂蛹虽然能较好提高SOD活性,抑制肾脏系数的升高,但不能降低血肌酐水平;蜂王浆虽然能降低BUN和肾组织中MDA水平,但会升高尿Cr,肾脏系数升高,且对小鼠无明显的抗氧化活力的提升。本发明所述蜂产品配方能显著增加肾组织中GSH的含量,提高SOD活力,降低SCr、BUN,起到对肾脏的保护效果。
通过与使用细胞保护剂—氨磷汀效果的对比,本发明所述蜂产品合剂配方能降低谷胱甘肽的消耗,提高抗氧化酶的活力,降低了血、尿中的肌酐和尿素氮水平,修复肾脏损伤,从而对肾脏起到了较好的保护作用。
Claims (3)
1.一种蜂产品胶囊的制备方法,其特征在于,具体包括以下步骤:
(1)将新鲜的刚从蜂巢取出的蜂蛹立即放入-80℃速冻;
(2)将胶体磨先与冰水循环冷却连接好,然后把速冻蜂蛹过胶体磨得到蜂蛹匀浆液;
(3)将蜂蛹匀浆液、蜂王浆、蜂蜜按质量比5:3:2的比例加入胶体磨中均质,得到蜂合剂;
(4)在蜂合剂加入辅料麦芽糊精和硬脂酸镁放入真空冷冻干燥机中冻干,将冻干粉过筛、灭菌、装入胶囊壳得到蜂产品胶囊。
2.根据权利要求1所述蜂产品胶囊的制备方法,其特征在于:步骤(4)中麦芽糊精的加入量为蜂合剂质量的50%,硬脂酸镁的加入量为蜂合剂质量的1%。
3.根据权利要求1所述蜂产品胶囊的制备方法,其特征在于:步骤(4)冻干的条件为:温度为-80℃,直至完全冻干成粉末,降温速率为快速降温,降至目标温度。
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