CN110734361A - 一种白皮杉醇的制备方法及应用 - Google Patents
一种白皮杉醇的制备方法及应用 Download PDFInfo
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- CDRPUGZCRXZLFL-OWOJBTEDSA-N piceatannol Chemical compound OC1=CC(O)=CC(\C=C\C=2C=C(O)C(O)=CC=2)=C1 CDRPUGZCRXZLFL-OWOJBTEDSA-N 0.000 title claims abstract description 79
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Abstract
本发明属于生物技术领域,具体涉及一种白皮杉醇的制备方法及应用,所述制备方法包括将百香果清洗后选取果肉,葡萄和蓝莓清洗后与百香果的果肉一起进行粉碎、过滤,得滤汁和滤渣,分别对滤汁和滤渣进行微波‑丙酮协同提取,合并提取液,减压浓缩,将浓缩液过硅胶柱,无水丙酮洗脱,冷冻干燥或二氧化碳超临界萃取,即得。本发明提供的提取方法,分别对百香果、葡萄和蓝莓的汁和渣进行提取,大大提高了产品收率,通过微波和丙酮协同提取的方法,进一步提高了产品收率,白皮杉醇的提取量达到2200mg/kg,且该制备方法耗时短,能耗低,适于工业化生产。
Description
技术领域
本发明属于生物技术领域,具体涉及一种白皮杉醇的制备方法及应用。
背景技术
白皮杉醇是天然提取物芪类物质的一种,是白藜芦醇3位羟基化类似物。药理学研究显示白皮杉醇有许多的药理活性,有抗癌、抗细胞增殖、抗炎、免疫调节抗脂质氧化、抗菌等多种药理活性。研究表明,白皮杉醇还是良好的抗氧化剂和心血管保护剂。随着开发的不断深入,白皮杉醇在保健品、化妆品以及药品等领域得到广泛的应用和认可。
百香果亦被称为鸡蛋果,果实球形或长圆球形,内有黄色果汁和黑色种子,含有蛋白质、脂肪、糖类、多种维生素、氨基酸和白皮杉醇等,营养丰富,酸甜可口,具有柠檬、柑、桔、橙子、草莓、酸梅、菠萝、石榴、香蕉、等水果香味,素有“饮料之王”、“水果味精”的美称,是生产果汁饮料的重要原料和添加剂。可以加工成果汁、果露、果酱、果冻、口含片,主要以加工成果汁食用为主,可制成芳香可口的饮料,还可用来添加在其他饮料中以提高饮料的品质。另外,在葡萄、蓝莓、甘蔗等植物中均发现白皮杉醇的存在。
目前,从水果中提取白皮杉醇的专利文献较少,且已有的从百香果中提取白皮杉醇的方法,其提取的白皮杉醇的含量较低。
发明内容
本发明的目的在于:提供一种成本低,步骤简单,含量高的白皮杉醇的制备方法,以解决上述背景技术中提出的问题。
为达到上述技术目的,本发明的技术方案:
一种白皮杉醇的制备方法,包括以下步骤:
(1)选取百香果、葡萄和蓝莓,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向所述滤汁中加入60-80%丙酮,微波提取6-10min,冷却至室温,得提取液;
(3)向所述滤渣中加入80-100%丙酮,微波提取4-6min,冷却至室温,过滤,得粗提液;
(4)将所述提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将所述浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将所述洗脱液进行冷冻干燥得白皮杉醇粉剂,或将所述洗脱液进行二氧化碳超临界萃取,得白皮杉醇液体。
作为一种改进,步骤(1)中,所述百香果、葡萄和蓝莓的重量比为6-8:1-2:1-2。
作为一种改进,步骤(2)中,所述滤汁与所述60-80%丙酮的重量比为1:6-8。
作为一种改进,步骤(3)中,所述滤渣与所述80-100%丙酮的重量比为1:4-6。
作为一种改进,步骤(2)和(3)中,所述微波的功率为200-250W。
作为一种改进,步骤(4)中,所述浓缩液的体积为所述提取液和粗提液合并液体积的1/20-1/15。
由于采用上述技术方案,本发明的有益效果:
本发明提供的白皮杉醇的制备方法,其操作简单,原料成本低,分别对百香果、葡萄和蓝莓的汁和渣进行提取,大大提高了产品收率,通过微波和丙酮协同提取的方法,进一步提高了产品收率,白皮杉醇的提取量达到2200mg/kg,且该制备方法耗时短,能耗低,适于工业化生产。
具体实施方式
下面结合具体实施方式对本发明作进一步的详细描述。但不应将此理解为本发明上述主题的范围仅限于以下的实施例,凡基于本发明内容所实现的技术均属于本发明的范围。
实施例1
一种白皮杉醇的制备方法,包括以下步骤:
(1)选取百香果8kg、葡萄1kg和蓝莓1kg,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向滤汁中加入6倍量的80%丙酮,在250W功率下,微波提取6min,冷却至室温,得提取液;
(3)向滤渣中加入6倍量的80%丙酮,在250W功率下,微波提取6min,冷却至室温,过滤,得粗提液;
(4)将提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将洗脱液于-40℃下进行冷冻干燥得白皮杉醇粉剂,或将洗脱液在45℃、30Mpa下进行二氧化碳超临界萃取3h,得白皮杉醇液体;经检测,白皮杉醇粉剂中白皮杉醇的含量为22.16g,从百香果、葡萄和蓝莓中的提取量为2216mg/kg;白皮杉醇液体中白皮杉醇的含量为22.08g,从百香果、葡萄和蓝莓中的提取量为2208mg/kg。
实施例2
一种白皮杉醇的制备方法,包括以下步骤:
(1)选取百香果7kg、葡萄1.5kg和蓝莓1.5kg,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向滤汁中加入7倍量的70%丙酮,在250W功率下,微波提取8min,冷却至室温,得提取液;
(3)向滤渣中加入5倍量的90%丙酮,在250W功率下,微波提取5min,冷却至室温,过滤,得粗提液;
(4)将提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将洗脱液于-40℃下进行冷冻干燥得白皮杉醇粉剂,或将洗脱液在50℃、35Mpa下进行二氧化碳超临界萃取2h,得白皮杉醇液体;经检测,白皮杉醇粉剂中白皮杉醇的含量为22.31g,从百香果、葡萄和蓝莓中的提取量为2231mg/kg;白皮杉醇液体中白皮杉醇的含量为22.28g,从百香果、葡萄和蓝莓中的提取量为2228mg/kg。
实施例3
一种白皮杉醇的制备方法,包括以下步骤:
(1)选取百香果6kg、葡萄2kg和蓝莓2kg,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向滤汁中加入8倍量的60%丙酮,在250W功率下,微波提取10min,冷却至室温,得提取液;
(3)向滤渣中加入4倍量的100%丙酮,在250W功率下,微波提取4min,冷却至室温,过滤,得粗提液;
(4)将提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将洗脱液于-40℃下进行冷冻干燥得白皮杉醇粉剂,或将洗脱液在40℃、25Mpa下进行二氧化碳超临界萃取4h,得白皮杉醇液体;经检测,白皮杉醇粉剂中白皮杉醇的含量为22.09g,从百香果、葡萄和蓝莓中的提取量为2209mg/kg;白皮杉醇液体中白皮杉醇的含量为22.05g,从百香果、葡萄和蓝莓中的提取量为2205mg/kg。
实施例4
一种白皮杉醇的制备方法,包括以下步骤:
(1)选取百香果7kg、葡萄2kg和蓝莓1kg,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向滤汁中加入7倍量的80%丙酮,在250W功率下,微波提取8min,冷却至室温,得提取液;
(3)向滤渣中加入5倍量的100%丙酮,在250W功率下,微波提取4min,冷却至室温,过滤,得粗提液;
(4)将提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将洗脱液于-40℃下进行冷冻干燥得白皮杉醇粉剂,或将洗脱液在50℃、30Mpa下进行二氧化碳超临界萃取3h,得白皮杉醇液体;经检测,白皮杉醇粉剂中白皮杉醇的含量为22.24g,从百香果、葡萄和蓝莓中的提取量为2224mg/kg;白皮杉醇液体中白皮杉醇的含量为22.19g,从百香果、葡萄和蓝莓中的提取量为2219mg/kg。
实施例5
一种白皮杉醇的制备方法,包括以下步骤:
(1)选取百香果6.5kg、葡萄1.5kg和蓝莓2kg,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向滤汁中加入8倍量的80%丙酮,在250W功率下,微波提取10min,冷却至室温,得提取液;
(3)向滤渣中加入5倍量的90%丙酮,在250W功率下,微波提取6min,冷却至室温,过滤,得粗提液;
(4)将提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将洗脱液于-40℃下进行冷冻干燥得白皮杉醇粉剂,或将洗脱液在45℃、35Mpa下进行二氧化碳超临界萃取2h,得白皮杉醇液体;经检测,白皮杉醇粉剂中白皮杉醇的含量为22.16g,从百香果、葡萄和蓝莓中的提取量为2216mg/kg;白皮杉醇液体中白皮杉醇的含量为22.13g,从百香果、葡萄和蓝莓中的提取量为2213mg/kg。
对照例
(1)取百香果10kg,用无菌离子水清洗干净,取百香果的果肉粉碎,过滤,得滤汁和滤渣;
(2)向滤汁中加入7倍量的80%丙酮,在250W功率下,微波提取8min,冷却至室温,得提取液;
(3)向滤渣中加入5倍量的100%丙酮,在250W功率下,微波提取6min,冷却至室温,过滤,得粗提液;
(4)将提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将洗脱液于-40℃下进行冷冻干燥得白皮杉醇粉剂,或将洗脱液在45℃、35Mpa下进行二氧化碳超临界萃取3h,得白皮杉醇液体;经检测,白皮杉醇粉剂中白皮杉醇的含量为6.25g,从百香果中的提取量为625mg/kg;白皮杉醇液体中白皮杉醇的含量为6.21g,从百香果中的提取量为621mg/kg。
本发明从百香果、葡萄和蓝莓中制得的白皮杉醇与对照例中单从百香果中制得的白皮杉醇,其提取量结果见表1:
表1
从表1中可以看出,从百香果、葡萄和蓝莓三种水果中制得的白皮杉醇的提取量可达2200mg/kg以上,而单从百香果中制得的白皮杉醇的提取量为620mg/kg以上,从百香果、葡萄和蓝莓三种水果中制得的白皮杉醇的提取量是单从百香果中制得的白皮杉醇的提取量的3.5倍,由此可见,通过本发明制得的白皮杉醇的产率较高。
本发明采用微波-丙酮协同进行粗提后,再通过硅胶柱进行分离纯化,以提高产品收率。微波提取技术是利用微波破碎细胞,溶剂进入细胞内部浸出有效成分的一种提取方法。它是基于微波加热的瞬时性、选择性和高效性,控制适宜的微波条件而实现的,植物细胞不同部位对微波能的吸收能力存在着差异,导致细胞内部水分蒸发汽化,使细胞内部压力增大而出现缺口,细胞内部有效成分析出。
白皮杉醇的应用
通过上述实施例制得的白皮杉醇液体,可作为食品添加剂添加到各种酒中,配制出对心血管疾病有良好预防作用的新型保健佐餐酒;或将白皮杉醇液体添加至食品中制成降脂、美容和减肥的天然保健食品;或将白皮杉醇液体添加至化妆品中,抑制黑色素生成,起到一定的美容作用。
通过上述实施例制得的白皮杉醇粉剂,可应用于抗白血病、预防和治疗动脉粥样硬化、心脑血管疾病等药物中。
以上所述本发明的具体实施方式,并不构成对本发明保护范围的限定。任何根据本发明的技术构思所做出的各种其他相应的改变与变形,均应包含在本发明权利要求的保护范围内。
Claims (8)
1.一种白皮杉醇的制备方法,其特征在于,包括以下步骤:
(1)选取百香果、葡萄和蓝莓,用无菌离子水清洗干净,取百香果的果肉与葡萄和蓝莓一起粉碎,过滤,得滤汁和滤渣;
(2)向所述滤汁中加入60-80%丙酮,微波提取6-10min,冷却至室温,得提取液;
(3)向所述滤渣中加入80-100%丙酮,微波提取4-6min,冷却至室温,过滤,得粗提液;
(4)将所述提取液和粗提液合并,减压浓缩,得浓缩液;
(5)将所述浓缩液通过硅胶柱,并用无水丙酮洗脱,得洗脱液;
(6)将所述洗脱液进行冷冻干燥得白皮杉醇粉剂,或将所述洗脱液进行二氧化碳超临界萃取,得白皮杉醇液体。
2.根据权利要求1所述的白皮杉醇的制备方法,其特征在于,步骤(1)中,所述百香果、葡萄和蓝莓的重量比为6-8:1-2:1-2。
3.根据权利要求1所述的白皮杉醇的制备方法,其特征在于,步骤(2)中,所述滤汁与所述60-80%丙酮的重量比为1:6-8。
4.根据权利要求1所述的白皮杉醇的制备方法,其特征在于,步骤(3)中,所述滤渣与所述80-100%丙酮的重量比为1:4-6。
5.根据权利要求1所述的白皮杉醇的制备方法,其特征在于,步骤(2)和(3)中,所述微波的功率为200-250W。
6.根据权利要求1所述的白皮杉醇的制备方法,其特征在于,步骤(4)中,所述浓缩液的体积为所述提取液和粗提液合并液体积的1/20-1/15。
7.根据权利要求1所述的白皮杉醇的制备方法制得的白皮杉醇粉剂在抗白血病药物中的应用。
8.根据权利要求1所述的白皮杉醇的制备方法制得的白皮杉醇液体在保健品或化妆品中的应用。
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