CN107281223A - 水解胎盘的方法、用该方法获得的超滤液及胎盘干燥产物 - Google Patents
水解胎盘的方法、用该方法获得的超滤液及胎盘干燥产物 Download PDFInfo
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Abstract
本发明提供水解胎盘的方法及用该方法获得的超滤液及胎盘干燥产物,称取胎盘冲洗干净得到清洗胎盘;将清洗胎盘剪成小块,在捣碎机中绞碎得到粉碎胎盘;向粉碎胎盘中加入小分子团水,放置于无菌不锈钢器皿中密封,于‑20℃或以下的温度速冻过夜,置于4℃‑10℃的环境完全融化,重复速冻与融化操作,得到水解液;将水解液离心获得上清液及沉淀物;将上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤获得分子量小于10KD的漏出液;将漏出液从1KD超滤柱中超滤后获得分子量在1KD‑10KD的超滤液;获得的沉淀物在56℃‑65℃的温度下真空干燥、超微粉碎,得到的胎盘干燥产物游离激素基本被去除,降低了传统“紫河车”制品干扰人体正常内分泌功能的问题。
Description
技术领域
本发明涉及生物技术领域,且特别是涉及水解胎盘的方法及用该方法获得的超滤液及胎盘干燥产物。
背景技术
胎盘又叫胞衣、衣胞、胎衣、胎膜。胎盘中药名为紫河车,具有调节人体机能,增强抵抗力,大补元气精血的作用。性温,味甘、咸,归心、肺、肾经。现代医学研究发现,胎盘含蛋白质、糖、钙、维生素、免疫因子、女性激素、助孕酮、类固醇激素、促性腺激素、促肾上腺皮质激素等,能促进乳腺、子宫、阴道、睾丸的发育,对甲状腺也有促进作用。对治疗子宫发育不全、子宫萎缩、子宫肌炎、机能性减退、无月经、子宫出血、乳汁缺乏症等,均有显着疗效;对肺结核、支气管哮喘、贫血等亦有良效;研末口服或灌肠可预防麻疹或减轻症状;对门静脉性肝硬化腹水及血吸虫性晚期肝硬化腹水也有一定的疗效。
然而传统中药“紫河车”是使用新鲜人胎盘经烘焙干燥后整体入药,或蒸煮成药,其存在如下技术缺陷:
1)、处理温度达到或接近100℃,大量生物活性物质被破坏,有益的生物效用大大降低;
2)、没有进行任何成分分离,残留有游离激素,存在干扰人体正常内分泌功能的潜在风险。
因此现有技术还有待于改进和发展。
发明内容
有鉴于此,本发明提供水解胎盘的方法及用该方法获得的超滤液及胎盘干燥产物,旨在解决现有技术中的采用强酸强碱来水解的做法导致引入污染物和其它有害因子以及产物含有含游离激素容易干扰人体正常内分泌功能的问题。
本发明解决技术问题所采用的技术方案如下:
水解胎盘的方法,包括以下步骤:
步骤1、称取新鲜或者冷冻胎盘,于洁净无菌环境下剔除筋膜及大血管,在4℃-10℃的无菌冰水中冲洗干净,得到清洗胎盘;
步骤2、将步骤1所得的清洗胎盘剪成小块,在捣碎机中绞碎,得到粉碎胎盘;
步骤3、向步骤2所得的粉碎胎盘中加入与步骤1中的新鲜或者冷冻胎盘等重的小分子团水,放置于无菌不锈钢器皿中,密封,于-20℃或以下的温度速冻过夜,然后置于4℃-10℃的环境下完全融化,反复速冻与融化操作多次,得到水解液;
步骤4、将步骤3中得到的水解液于离心机中进行离心30分钟-60分钟,获得上清液及沉淀物;其中,所述离心机的离心速度至少为3500RPM;
步骤5、将步骤4获得的上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤后,弃穿透液留漏出液,获得分子量小于10KD的漏出液;将所述漏出液从1KD超滤柱中超滤后,弃漏出液留穿透液,获得分子量在1KD-10KD的超滤液;
步骤6、将步骤4获得的沉淀物在56℃-65℃的温度下真空干燥、超微粉碎,得到胎盘干燥产物。
所述的水解胎盘的方法,其中,所述步骤1中的冷冻胎盘需预先在4℃-10℃的环境下完全解冻。
所述的水解胎盘的方法,其中,所述步骤2中得到的粉碎胎盘的粒度为60目-120目。
所述的水解胎盘的方法,其中,所述步骤3中的小分子团水的PH值为9.6-12.5。
所述的水解胎盘的方法,其中,所述步骤3中的反复速冻与融化操作的次数为3次。
所述的水解胎盘的方法,其中,所述步骤6中的沉淀物在60℃的温度下真空干燥。
所述的水解胎盘的方法,其中,所述步骤1中的胎盘为人或者动物的胎盘。
一种超滤液,通过如上所述的水解胎盘的方法制备获得。
一种胎盘干燥产物,通过如上所述的水解胎盘的方法制备获得。
与现有技术相比,本发明具有以下优点:
首先,本发明通过强极性的小分子团水可以强力水解胎盘的生物大分子,避免了传统技术中由于采用强酸强碱来水解的做法导致引入污染物和其它有害因子;同时结合了-20℃与4℃的反复多次冻融,避免了高温对生物活性物质的破坏,最大限度地保留了胎盘中生物分子的活性;得到的胎盘干燥产物可以替代中药材“紫河车”入药。强极性的小分子团水可以强力溶出胎盘组织中的游离激素,经离心去上清,干燥沉淀物得到的胎盘干燥产物中的游离激素基本被去除,大大降低了传统“紫河车”制品中残留游离激素而干扰人体正常内分泌功能的问题;
其次,配合使用10KD和1KD两个超滤柱进行分子筛截留操作,既去除了较难吸收的、分子量大于10KD的物质,也去除了主要含游离激素的、分子量小于1KD的物质,获得的超滤液富含分子量1-10KD小分子核酸及多肽,可广泛用于生物制药、美容护肤及保健食品。
具体实施方式
为使本发明的目的、技术方案及优点更加清楚、明确,以下结合实施例对本发明进一步详细说明。应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。
本发明提供了水解胎盘的方法,包括以下步骤:
步骤1、称取新鲜或者冷冻胎盘,于洁净无菌环境下剔除筋膜及大血管,在4℃-10℃的无菌冰水中冲洗干净,得到清洗胎盘;
步骤2、将步骤1所得的清洗胎盘剪成小块,在捣碎机中绞碎,得到粉碎胎盘;
步骤3、向步骤2所得的粉碎胎盘中加入与步骤1中的新鲜或者冷冻胎盘等重的小分子团水,放置于无菌不锈钢器皿中,密封,于-20℃或以下的温度速冻过夜,然后置于4℃-10℃的环境下完全融化,反复速冻与融化操作多次,得到水解液;
步骤4、将步骤3中得到的水解液于离心机中进行离心30分钟-60分钟,获得上清液及沉淀物;其中,所述离心机的离心速度至少为3500RPM;其中,游离基素主要被留在了上清液里,沉淀物的游离激素基本被去除。
步骤5、将步骤4获得的上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤后,弃穿透液留漏出液,获得分子量小于10KD的漏出液;将所述漏出液从1KD超滤柱中超滤后,弃漏出液留穿透液,获得分子量在1KD-10KD的超滤液;
步骤6、将步骤4获得的沉淀物在56℃-65℃的温度下真空干燥、超微粉碎,得到胎盘干燥产物。
其中,步骤1中的冷冻胎盘需预先在4℃-10℃的环境下完全解冻。
所述步骤2中得到的粉碎胎盘的粒度为60目-120目。
下面给出本发明水解胎盘的方法的几个实施例。
实施例1
步骤1、称取人的新鲜胎盘,于洁净无菌环境下剔除筋膜及大血管,在4℃的无菌冰水中冲洗干净,得到清洗胎盘;其中,所述粉碎胎盘的粒度为60目;
步骤2、将步骤1所得的清洗胎盘剪成小块,在捣碎机中绞碎,得到粉碎胎盘;
步骤3、向步骤2所得的粉碎胎盘中加入与步骤1中的新鲜胎盘等重的pH为12.5的小分子团水,放置于无菌不锈钢器皿中,密封,于-20℃的温度速冻过夜,然后置于4℃的环境下完全融化,反复速冻与融化操作3次,得到水解液;
其中,所述小分子团水是由3-6个水分子缔结而成,其运动速度快,具有极强的渗透力、扩散力、溶解力。小分子团水的PH值越高,水解的效率就越高。
步骤4、将步骤3中得到的水解液于离心机中进行离心30分钟,获得上清液及沉淀物;其中,所述离心机的离心速度为3500RPM;
步骤5、将步骤4获得的上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤后,弃穿透液留漏出液,获得分子量小于10KD的漏出液;将所述漏出液从1KD超滤柱中超滤后,弃漏出液留穿透液,获得分子量在1KD-10KD的超滤液;
步骤6、将步骤4获得的沉淀物在56℃的温度下真空干燥、超微粉碎,得到胎盘干燥产物。
实施例2
步骤1、称取动物的冷冻胎盘,预先在4℃的环境下完全解冻,然后于洁净无菌环境下剔除筋膜及大血管,在10℃的无菌冰水中冲洗干净,得到清洗胎盘;
步骤2、将步骤1所得的清洗胎盘剪成小块,在捣碎机中绞碎,得到粉碎胎盘;其中,所述粉碎胎盘的粒度为120目;
步骤3、向步骤2所得的粉碎胎盘中加入与步骤1中的冷冻胎盘等重的pH为10的小分子团水,放置于无菌不锈钢器皿中,密封,于-25℃的温度速冻过夜,然后置于7℃的环境下完全融化,反复速冻与融化操作4次,得到水解液;
步骤4、将步骤3中得到的水解液于离心机中进行离心45分钟,获得上清液及沉淀物;其中,所述离心机的离心速度为4000RPM;
步骤5、将步骤4获得的上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤后,弃穿透液留漏出液,获得分子量小于10KD的漏出液;将所述漏出液从1KD超滤柱中超滤后,弃漏出液留穿透液,获得分子量在1KD-10KD的超滤液;
步骤6、将步骤4获得的沉淀物在60℃的温度下真空干燥、超微粉碎,得到胎盘干燥产物。
实施例3
步骤1、称取动物的冷冻胎盘,预先在10℃的环境下完全解冻,然后于洁净无菌环境下剔除筋膜及大血管,在5℃的无菌冰水中冲洗干净,得到清洗胎盘;
步骤2、将步骤1所得的清洗胎盘剪成小块,在捣碎机中绞碎,得到粉碎胎盘;其中,所述粉碎胎盘的粒度为80目;
步骤3、向步骤2所得的粉碎胎盘中加入与步骤1中的冷冻胎盘等重的pH为9.6的小分子团水,放置于无菌不锈钢器皿中,密封,于-30℃的温度速冻过夜,然后置于10℃的环境下完全融化,反复速冻与融化操作5次,得到水解液;
步骤4、将步骤3中得到的水解液于离心机中进行离心60分钟,获得上清液及沉淀物;其中,所述离心机的离心速度为5000RPM;
步骤5、将步骤4获得的上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤后,弃穿透液留漏出液,获得分子量小于10KD的漏出液;将所述漏出液从1KD超滤柱中超滤后,弃漏出液留穿透液,获得分子量在1KD-10KD的超滤液;
步骤6、将步骤4获得的沉淀物在65℃的温度下真空干燥、超微粉碎,得到胎盘干燥产物。
与现有技术相比,本发明具有以下优点:
首先,本发明通过强极性的小分子团水可以强力水解胎盘的生物大分子,避免了传统技术中由于采用强酸强碱来水解的做法导致引入污染物和其它有害因子;同时结合了-20℃与4℃的反复多次冻融,避免了高温对生物活性物质的破坏,最大限度地保留了胎盘中生物分子的活性;得到的胎盘干燥产物可以替代中药材“紫河车”入药,强极性的小分子团水可以强力溶出胎盘组织中的游离激素,经离心去上清,干燥沉淀物得到的胎盘干燥产物中的游离激素基本被去除,大大降低了传统“紫河车”制品中残留游离激素而干扰人体正常内分泌功能的问题;
其次,配合使用10KD和1KD两个超滤柱进行分子筛截留操作,既去除了较难吸收的、分子量大于10KD的物质,也去除了主要含游离激素的、分子量小于1KD的物质,获得的超滤液富含分子量1-10KD小分子核酸及多肽,可广泛用于生物制药、美容护肤及保健食品。
本发明还提供了一种超滤液,通过如上具体实施例的水解胎盘的方法制备获得。
本发明还提供了一种胎盘干燥产物,通过如上具体实施例的水解胎盘的方法制备获得。
用本发明的超滤液分别做皮肤刺激性检测、急性眼刺激检测、皮肤变态反应检测实验,以证明其安全性,如下所示:
(1)皮肤刺激性检测
①选取健康成年白色家兔4只,体重2.4-2.8kg,雌雄兼用,控制动物室的温度在22-25℃,相对湿度为55-70%;
②实验前日,将家兔背部脊柱两侧毛剪掉,去毛范围左、右各约3cm×3cm。将0.5ml的的样品涂抹于左侧去毛皮肤上,范围2.5×2.5cm2,每天涂抹一次,连续涂抹14天,右侧皮肤作为对照。从第二天开始,每天涂抹前去毛,用清水清洗,一小时后观察皮肤反应。按皮肤刺激强度分级标准判定受试物对皮肤的刺激强度,实验结果如表1所示。
表1
受试物对家兔多次皮肤刺激性试验结果
(2)急性眼刺激检测
①选取健康成年白色家兔3只,体重2.2-2.5kg,雌雄兼用,控制动物室的温度在22-25℃,相对湿度为55-70%;
②将0.1ml样品滴入家兔一侧眼结膜囊内,立刻闭合兔眼1s,至第30s用足量、流速较快的生理盐水冲洗30s;另一侧眼不处理作自身对照。分别在1h、24h、48h、72h对家兔眼镜进行检查(24h后使用荧光素钠)。按眼刺激性反应分级标准判定受试物对眼的刺激强度。滴眼后1h各家兔受试眼可见眼结构膜轻度充血,24h后恢复正常,实验结果如表2所示。
表2
受试物对家兔急性眼刺激性试验结果
(3)皮肤变态反应检测
①选取健康成年白色豚鼠36只,体重200g-300g,雌雄各半,控制动物室的温度在22-25℃,相对湿度为55-70%;
②采用局部封闭涂皮法,将豚鼠分成试验组(20只)和阴性对照组(16只),每组雌、雄性各半。
③诱导接触:试验前24h,将豚鼠背部左侧毛剃掉,范围约3cm×3cm,将0.2ml样品涂抹在受试豚鼠背部左侧去毛皮肤上,涂抹范围2cm×2cm,以二层纱布和一层玻璃纸覆盖,再以无刺激胶布封闭固定6h,然后去除受试物。第7天和第14天各用同样的方法重复一次。阴性对照组豚鼠不给予激发接触。
④激发接触:末次致敏后14天,按上述方法分别将样品涂于试验组豚鼠的背部右侧的去毛区,以同样的方法用样品对阴性对照组豚鼠进行激发接触,于除药后24h和48h后观察豚鼠背部右侧受试区皮肤反应。实验结果如表3所示。
表3
受试物对豚鼠皮肤变态反应试验结果
对本发明的胎盘水解超滤液做四十一种糖皮质激素检测实验,以证明其不含任何皮质激素,四十一种糖皮质激素的种类见表4所示。
表4
四十一种糖皮质激素
利用GB/T24800.2-2009,液相色谱/串联质谱法的测定方法对本发明的胎盘水解超滤液做四十一种糖皮质激素检测实验,结果显示均未检测出表4所示的四十一种糖皮质激素。
由此可知,本发明制备的胎盘水解超滤液不含任何皮质激素,长期使用不会伤害皮肤细胞,适合用于配制护肤精华液、各种膏霜乳液及面膜液等各种护肤产品。
最后说明的是,以上实施例仅用来说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的宗旨和范围,其均应涵盖在本发明的权利要求范围当中。
Claims (9)
1.水解胎盘的方法,其特征在于,包括以下步骤:
步骤1、称取新鲜或者冷冻胎盘,于洁净无菌环境下剔除筋膜及大血管,在4℃-10℃的无菌冰水中冲洗干净,得到清洗胎盘;
步骤2、将步骤1所得的清洗胎盘剪成小块,在捣碎机中绞碎,得到粉碎胎盘;
步骤3、向步骤2所得的粉碎胎盘中加入与步骤1中的新鲜或者冷冻胎盘等重的小分子团水,放置于无菌不锈钢器皿中,密封,于-20℃或以下的温度速冻过夜,然后置于4℃-10℃的环境下完全融化,反复速冻与融化操作多次,得到水解液;
步骤4、将步骤3中得到的水解液于离心机中进行离心30分钟-60分钟,获得上清液及沉淀物;其中,所述离心机的离心速度至少为3500RPM;
步骤5、将步骤4获得的上清液经定性滤纸抽滤后获得粗滤液,将粗滤液从10KD超滤柱中超滤后,弃穿透液留漏出液,获得分子量小于10KD的漏出液;将所述漏出液从1KD超滤柱中超滤后,弃漏出液留穿透液,获得分子量在1KD-10KD的超滤液;
步骤6、将步骤4获得的沉淀物在56℃-65℃的温度下真空干燥、超微粉碎,得到胎盘干燥产物。
2.如权利要求1所述的水解胎盘的方法,其特征在于,所述步骤1中的冷冻胎盘需预先在4℃-10℃的环境下完全解冻。
3.如权利要求1所述的水解胎盘的方法,其特征在于,所述步骤2中得到的粉碎胎盘的粒度为60目-120目。
4.如权利要求1所述的水解胎盘的方法,其特征在于,所述步骤3中的小分子团水的PH值为9.6-12.5。
5.如权利要求1所述的水解胎盘的方法,其特征在于,所述步骤3中的反复速冻与融化操作的次数为3次。
6.如权利要求1所述的水解胎盘的方法,其特征在于,所述步骤6中的沉淀物在60℃的温度下真空干燥。
7.如权利要求1所述的水解胎盘的方法,其特征在于,所述步骤1中的胎盘为人或者动物的胎盘。
8.一种超滤液,其特征在于,利用权利要求1-7任一项所述的水解胎盘的方法制备获得。
9.一种胎盘干燥产物,其特征在于,利用权利要求1-7任一项所述的水解胎盘的方法制备获得。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102488713A (zh) * | 2011-12-26 | 2012-06-13 | 重庆大学 | 一种制备羊胎盘素和羊胎盘水解胶原浓缩液的方法 |
CN104940100A (zh) * | 2015-06-19 | 2015-09-30 | 成都清科生物科技有限公司 | 一种人胎盘提取物及其制备方法和应用 |
JP2016222612A (ja) * | 2015-06-01 | 2016-12-28 | 昭和電工株式会社 | 化粧料または皮膚外用剤 |
-
2017
- 2017-06-29 CN CN201710515633.XA patent/CN107281223A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102488713A (zh) * | 2011-12-26 | 2012-06-13 | 重庆大学 | 一种制备羊胎盘素和羊胎盘水解胶原浓缩液的方法 |
JP2016222612A (ja) * | 2015-06-01 | 2016-12-28 | 昭和電工株式会社 | 化粧料または皮膚外用剤 |
CN104940100A (zh) * | 2015-06-19 | 2015-09-30 | 成都清科生物科技有限公司 | 一种人胎盘提取物及其制备方法和应用 |
Non-Patent Citations (2)
Title |
---|
佟志清等: "胎盘肽注射液的研制及治疗外阴白斑的疗效", 《药学实践杂志》 * |
李硕等: "酶解法和超滤法获得羊胎盘提取物对人脐静脉内皮细胞增殖影响差异", 《解剖科学进展》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109276531A (zh) * | 2018-11-09 | 2019-01-29 | 东阿阿胶股份有限公司 | 一种含有驴胎盘提取物的面膜及其制备工艺 |
CN110540575A (zh) * | 2019-06-06 | 2019-12-06 | 殷洪波 | 一种益生菌发酵去激素胎盘复合多肽的制备方法 |
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