CN115181152A - 一种鸭胆中提取鹅去氧胆酸的方法 - Google Patents
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Abstract
本发明涉及一种鸭胆中提取鹅去氧胆酸的方法。本方法的基本特征在于,其鸭胆汁通过酰胺键水解酶,在4h内基本完全水解TCDCA成CDCA;通过少量片碱水解蛋白质;然后通过溶剂萃取CDCA,再成盐,将胆汁中的大部分胆酸类物质全部分离。其水解速度快、彻底、提取率高、操作简单、污染小,适合于工业化,有效解决了传统提取方法中使用大量片碱、水解困难等问题。
Description
技术领域
本发明属于医药技术领域,涉及从鸭胆中提取鹅去氧胆酸的新方法。
背景技术
鹅去氧胆酸(CDCA),中文名:鹅去氧胆酸,英文名Chenodeoxycholic acid。化学名: 3α,7α-二羟基-5β-胆烷酸。其结构如下:
鹅去氧胆酸(CDCA)主要作用是降低胆汁内胆固醇的饱和度,也是熊去氧胆酸(UDCA) 合成的主要原料。鹅去氧胆酸目前主要来源是从鸡胆、鸭胆、猪胆中提取,在鸭胆胆汁中主要存在鹅去氧胆酸和海豹胆酸,二者均以与甘氨酸结合形式存在,仅有很少部分以游离态存在。
发明内容
本发明另一方面,提供了一种从鸭胆中提取鹅去氧胆酸的新方法。
本发明包含以下内容:
鸭胆——>鸭胆汁——>酶解——>碱解——>过滤——>萃取——>成盐
本发明中所述的鸭胆为新鲜鸭胆或冷冻鸭胆。
本发明中所述的酶解中,酶为专属性的酰胺水解酶,酶添加量为胆汁的1.0-2.0%,反应 pH在4.0-6.0之间,反应温度在25-45℃之间,反应时间1-5h,调节pH的酸为稀硫酸、磷酸、洗盐酸或浓盐酸。优选酶添加量1.0%,反应pH在5.0-5.5之间,反应温度为30-35℃,反应时间为4h,pH调节优选浓盐酸。
本发明中所述的碱解中碱为NaOH、KOH,温度80-120℃,水解时间为4-14h。优选NaOH,反应温度95℃,碱量为胆汁重量的3%,水解时间优选8h。
本发明中所述的萃取所用溶剂为乙酸乙酯、乙酸丁酯、乙酸异丙酯、甲基异丁基酮、戊酮,酸化用硫酸、磷酸、稀盐酸或浓盐酸。优选溶剂为乙酸乙酯,酸化优选浓盐酸或磷酸
本发明中所述的成盐中所用的溶剂为乙酸乙酯、乙酸丁酯、乙酸异丙酯、甲基异丁基酮、戊酮、丙酮,有机胺为仲丁胺、叔丁胺、乙二胺、三乙醇胺,析晶时间1-4h;有机胺的添加量为胆汁质量的1.0%-2.0%。优选溶剂为乙酸乙酯,有机胺为仲丁胺,析晶时间2h,胺用量为胆汁质量的1.5%。
附图说明:
图1为实施例1中酶水解液RID谱图;
图2为实施例2中酶水解液RID谱图;
图3实施例1中成品胺盐RID谱图;
图4为实施例2中成品胺盐RID谱图。
具体实施方式
为了使本领域技术人员可以更好地理解本发明,以下通过具体实施例对本发明技术方案进行进一步说明。需要理解的是,下属实施例只为更好地说明本发明而给出,并不是对本发明内容的限制。
实施例1:
取冷冻鸭胆100g,切碎,加热至70℃完全溶解,胆汁降温至30℃,用浓盐酸调节pH至5.0,保持温度30-35℃,加入水解酶1g,继续调节pH保持在5.0-5.5,反应4h,检测转化率,其酶水解液RID谱图如图1。加入3g片碱,升温至95℃,反应8h,降温,盐酸调节pH=2-3,过滤,加入200ml乙酸乙酯,水50ml,萃取2h,静置,分去水相,有机相加入2g三乙醇胺,搅拌析晶2h。过滤,烘干,得粗鹅去氧胆酸胺盐3.7g,鹅去氧胆酸提取率81.4%,RID检测,其成品胺盐RID谱图如图3。
实施例2:
取新鲜鸭胆500g,放出胆汁,胆汁加热至30℃,用浓盐酸调节pH至5.0,保持温度30-35℃,加入水解酶5g,继续调节pH保持在5.0-5.5,反应4h,检测转化率,其酶水解液RID谱图如图2。继续保温反应1h,用盐酸调节pH至2-3,固体加入15g片碱,升温至100℃,反应8h,降温,加入浓盐酸,调节pH至3-4,过滤,滤饼加入600ml乙酸丁酯和200ml水,萃取2h,静置,分去水相,有机相加入10g叔丁胺,搅拌析晶2h。过滤,烘干,得粗鹅去氧胆酸胺盐30.6g,鹅去氧胆酸提取率91.8%,RID检测,其成品胺盐RID谱图如图4。
实施例3:
取冷冻鸭胆2000g,切碎,加热至70℃,过滤胆皮,胆汁降温至30℃,用浓盐酸调节pH 至5.0,保持温度30-35℃,加入水解酶20g,继续调节pH保持在5.0-5.5,反应4h,检测转化率,继续保温反应1h,用盐酸调节pH至2-3,固体加入60g片碱,升温至95℃,反应8h,降温,加入浓盐酸,调节pH至3-4,过滤,滤饼加入2000ml甲基异丁基酮和500ml水,萃取2h,静置,分去水相,有机相加入30g仲丁胺,搅拌析晶2h。过滤,烘干,得粗鹅去氧胆酸胺盐89.2g,鹅去氧胆酸提取率98.2%。
Claims (9)
1.一种鸭胆中提取鹅去氧胆酸的方法,其基本特征在于,鸭胆汁通过酶水解,少量的碱解,然后通过一定比例的溶剂和水萃取CDCA,再通过有机胺成胺盐。
2.如权利要求1所述,酶水解过程是将胆汁调节pH和温度,加入适量的水解酶,水解过程用酸调节pH至反应结束。
3.上述权利要求中所述的酶解其基本特征在于,酶添加量为胆汁的1.0%-2.0%,反应pH在4.0-6.0之间,反应温度在25-45℃之间,反应时间1-5h。
4.如权利要求1所述的碱解过程是将酶解结束后的胆汁加入碱,高温水解一段时间。
5.上述权利要求中所述的碱为NaOH、KOH,水解温度为80-120℃,水解时间为4-14h。
6.权利要求1所述萃取所用的溶剂为乙酸乙酯、乙酸丁酯、乙酸异丙酯、甲基异丁基酮、戊酮,所用溶剂倍数为1-5倍。
7.上述权利要求中溶剂和水的比例为3:1—10:1。
8.权利要求1所述的胺盐制备过程方法为,在溶解了胆酸的溶剂中,加入一定比例的有机胺,搅拌析晶,过滤。
9.上述权利要求中所述的有机胺为叔丁胺、仲丁胺、三乙醇胺、二乙胺等;成盐用溶剂为乙酸乙酯、乙酸丁酯、乙酸异丙酯、甲基异丁基酮、丙酮、戊酮、丁酮;析晶时间1-4h;有机胺的添加量为胆汁质量的1.0%-2.0%。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0092073A1 (en) * | 1982-04-15 | 1983-10-26 | SCHWARZ ITALIA S.p.A. | Magnesium salt of chenodeoxycholic acid and ursodeoxycholic acid, the process for its preparation, and therapeutic compositions which contain it as active principle |
CN111334553A (zh) * | 2020-03-26 | 2020-06-26 | 山东中京生物科技有限公司 | 一种酶法胆膏生产工艺 |
CN112062802A (zh) * | 2019-06-11 | 2020-12-11 | 四川澄华生物科技有限公司 | 鹅去氧胆酸乙酸丁酯提取液和制备方法、以及鹅去氧胆酸铵盐和鹅去氧胆酸的制备方法 |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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EP0092073A1 (en) * | 1982-04-15 | 1983-10-26 | SCHWARZ ITALIA S.p.A. | Magnesium salt of chenodeoxycholic acid and ursodeoxycholic acid, the process for its preparation, and therapeutic compositions which contain it as active principle |
CN112062802A (zh) * | 2019-06-11 | 2020-12-11 | 四川澄华生物科技有限公司 | 鹅去氧胆酸乙酸丁酯提取液和制备方法、以及鹅去氧胆酸铵盐和鹅去氧胆酸的制备方法 |
CN111334553A (zh) * | 2020-03-26 | 2020-06-26 | 山东中京生物科技有限公司 | 一种酶法胆膏生产工艺 |
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