CN112546077A - 一种桑黄中降血糖物质的联合提取方法及应用 - Google Patents
一种桑黄中降血糖物质的联合提取方法及应用 Download PDFInfo
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- CN112546077A CN112546077A CN202011494218.9A CN202011494218A CN112546077A CN 112546077 A CN112546077 A CN 112546077A CN 202011494218 A CN202011494218 A CN 202011494218A CN 112546077 A CN112546077 A CN 112546077A
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Abstract
本发明涉及从天然产物中提取活性物质的技术领域,具体涉及一种从人工栽培的真菌‑桑黄中提取具有降血糖作用的物质的方法,包括从桑黄子实体中醇提取小分子物质后再水提取糖类物质。与现有技术相比,本发明提供的两步联合提取法具有较高的提取率,且方法工艺简单,安全环保,成本低廉,可操作性强,易于实际生产应用。同时,本发明提取的小分子物质具有强烈的α‑葡萄糖苷酶抑制活性。
Description
技术领域
本发明涉及从天然产物中提取活性物质的技术领域,具体涉及一种从人工栽培的真菌-桑黄中提取具有降血糖作用的物质的方法。
背景技术
糖尿病是一种以高血糖为特征的慢性代谢疾病,在全世界发病率高,更是成为全球三大慢性非传染疾病之一,官方数据显示到2025年,全球糖尿病发病人群将高达3.33亿人次。糖尿病的病因是由于受损的胰腺β细胞导致胰岛素分泌不足或者胰岛素抵抗等造成的,目前常通过修复受损的β细胞、增加胰岛素敏感性、延缓糖类的消化吸收、改善糖代谢、增加周围组织对葡萄糖的利用等途径进行干预。
桑黄(Phellinusigniarius)是一种寄生于桑树树干的大型珍贵药用真菌,属担子菌纲、多孔菌目、多孔菌科、针层孔属的真菌。桑黄的药用最早记载于《本草纲目》,传统具有化瘀、止血等作用。现代药理学研究表明,桑黄具有抗肿瘤、抗氧化、降血脂、增强免疫力等功效,是国际公认的最佳抗癌真菌之一。桑黄主要的活性成分有小分子的三萜酸和黄酮类物质,以及大分子的多糖类物质等[1],也已有专利和非专利文献报道对桑黄中这些物质进行了提取和纯化,黄酮的提取率在1.5~8.45%之间[2-5],其中以李善姬等[2]的工艺中黄酮提取率最高;多糖的提取率在1.1~5.5%之间[6-7]。比如公开号为CN110882285A、公开日为2020年03月17日的中国发明申请公开了一种通过MgO和四氢呋喃同时提取桑黄和多酚的方法[8]。又如公开号为CN110407949A、公开日为2019年11月05日的中国发明申请公开了一种从桑黄中提取和纯化多糖的方法[9]。但现有文献对提取物的功能活性研究还较少,桑黄中降血糖成分的研究多集中于多糖类成分,对于黄酮类等小分子物质的降血糖研究还较少。
α-葡萄糖苷酶是把碳水化合物水解成正常生理活动所需的葡萄糖的关键酶,抑制α-葡萄糖苷酶活性可以延迟碳水化合物的消化和吸收,从而减缓了血糖的快速升高,增强对胰岛素刺激作用的敏感性并减少对胰腺刺激作用,因此目前已经开发了多种α-葡萄糖苷酶抑制剂的药物。但是合成的药物具有一定的毒副作用,且价格较昂贵,因此,已经开始逐渐从天然产物中筛选α-葡萄糖苷酶抑制剂。
发明内容
针对现有技术中桑黄提取物提取率低、对提取物的功能活性研究还较少,桑黄中降血糖成分的研究多集中于多糖类成分,对于黄酮类等小分子物质的降血糖研究还较少等一系列问题,本发明的目的是提供一种从人工栽培的桑黄中两步提取具有降血糖成分的方法,提高桑黄活性成分的提取率,同时提供桑黄提取物中黄酮类小分子的作用研究探索,为桑黄的进一步利用提供技术支持。
为实现本发明的发明目的,发明人提供如下技术方案:
本发明的桑黄采用人工栽培的桑黄子实体,采自浙江省农业科学院蚕桑研究所桑黄种植基地。
本发明首先提供了一种桑黄中降血糖物质的联合提取方法,包括如下步骤:
(1)取桑黄子实体,干燥、粉碎并过筛,备用;
(2)按照固液比为1:10~1:30,在桑黄子实体粉末中加入乙醇,在温度60~80℃下提取1~3h,重复提取数次,冷却,离心,取上清液1备用;
(3)上清液1先用微孔滤膜过滤得到微滤液1,过滤掉较大颗粒的悬浮物,再利用截留分子量为10KD的超滤膜在压力为0.15~0.25Mpa、温度为25~40℃的工作条件下对微滤液1进行超滤,得到超滤液1,超滤液1进行减压浓缩,回收其中的乙醇,浓缩液1最后经真空冷冻干燥即得到桑黄提取物1,
(4)为了充分利用桑黄中的活性成分,按照固液比1:20~1:40,在步骤(2)提取离心后剩余的残渣中加入水,在温度为90~100℃下提取1.5~2h,冷却,离心,取上清液2备用;
(5)上清液2先用微孔滤膜包过滤得到微滤液2,再利用截留分子量为1KD的复合超滤膜包在压力为0.1~0.15Mpa、温度为25~40℃的工作条件下对微滤液2进行超滤,取截留液,减压浓缩后再经真空冷冻干燥,得到冻干粉,此冻干粉为醇提后回收的水提物-桑黄提取物2,主要成分为多糖。
作为优选,根据本发明所述的一种桑黄中降血糖物质的联合提取方法,其中,所述的步骤(1)中桑黄子实体在50~70℃下干燥,粉碎后过30~60目筛。
作为优选,根据本发明所述的一种桑黄中降血糖物质的联合提取方法,其中,所述的步骤(2)中乙醇浓度为40~80%(体积比)。
作为优选,根据本发明所述的一种桑黄中降血糖物质的联合提取方法,其中,所述的步骤(3)中微孔滤膜采用聚醚砜微孔滤膜;超滤膜采用聚醚砜超滤膜。
作为优选,根据本发明所述的一种桑黄中降血糖物质的联合提取方法,其中,所述的步骤(5)中微孔滤膜包采用混合纤维素微孔滤膜包;复合超滤膜包采用聚醚砜-聚酰胺复合超滤膜包。
作为优选,根据本发明所述的一种桑黄中降血糖物质的联合提取方法,其中,所述的步骤(5)中超滤:回流液流回原液中,当截取液体积减少至原液初始体积的30%-35%,透过液速度明显降低时,停止超滤。
本发明还提供了桑黄提取物在制备α-葡萄糖苷酶抑制剂上的应用,所述的桑黄提取物为醇提取物。
发明人在筛选桑黄中抑制α-葡萄糖苷酶的成分时发现,桑黄的水提取成分对α-葡萄糖苷酶的抑制活性较低,表明多糖类的降血糖作用不是通过抑制α-葡萄糖苷酶的途径进行的;而桑黄60%的乙醇提取物对α-葡萄糖苷酶的抑制活性很强,表明桑黄中具有强烈抑制α-葡萄糖苷酶的物质存在,而这部分物质也可能发挥着降血糖作用。
因此,本发明首先以α-葡萄糖苷酶抑制活性为引导,第一步采用醇提法从桑黄中提取小分子物质,并通过超滤质谱技术筛选鉴定出具有α-葡萄糖苷酶抑制活性的成分;第二步对经过醇提取后的剩余残渣水提取处理得到桑黄的水提物,最后通过动物试验验证了两步提取物的降血糖作用。
本发明中,作为优选,所述的桑黄醇提取物按下述方法提取:
按照固液比为1:10~1:30,在桑黄子实体粉末中加入乙醇,在温度60~80℃下提取1~3h,重复提取数次,冷却,离心,取上清液1备用;
上清液1先用微孔滤膜过滤得到微滤液1,再利用截留分子量为10KD的超滤膜在压力为0.15~0.25Mpa、温度为25~40℃的工作条件下对微滤液进行超滤,得到超滤液1,超滤液1进行减压浓缩,回收其中的乙醇,浓缩液1最后经真空冷冻干燥即得到桑黄提取物1。
作为优选,本发明应用中,所述的微孔滤膜采用聚醚砜微孔滤膜;超滤膜采用聚醚砜超滤膜。
本发明通过超滤质谱技术筛选鉴定出桑黄醇提取物中具有α-葡萄糖苷酶抑制活性的成分,所述的桑黄醇提取物,其组成包括并不限于下列成分:
原儿茶酸、咖啡酸、二氢山柰酚、7,8-二羟基-4-甲基香豆素、山柰酚、橘霉素、桑色素D、咖啡豆醇、脱氢枞酸、桦木酸和熊果酸。
本发明中:
(1)α-葡萄糖苷酶抑制成分的提取
设立了四因素三水平的正交试验,称取桑黄适量,按表中的条件进行提取,提取结束后,进行离心,量取上清液体积并测定总黄酮含量,测定方法参考发明者已有专利中的方法(中国专利文献CN201310442734.0)。把上清液浓缩干燥,取干燥物适量,用DMSO溶液配成10g/L的溶液,再用缓冲溶液稀释100倍用于α-葡萄糖苷酶的抑制试验,测定方法参考发明者的文献[10]。对提取后的残渣进行了多糖的提取,提取条件为水提,料液比1∶20,在70℃下提取1小时,离心,量取上清液体积,并测定多糖含量,测定方法为苯酚硫酸法。
表1正交试验结果
从试验结果表1可知,总黄酮最佳的提取工艺条件为乙醇浓度40%,提取温度80℃,提取时间3h,料液比1∶30。对α-葡萄糖苷酶抑制率最高的最佳的提取工艺条件为乙醇浓度60%,提取温度70℃,提取时间3h,料液比1∶30。正交试验提取后的残渣的多糖提取率基本与黄酮提取率相反。为了实现黄酮和多糖联合的提取,可以选择黄酮和多糖提取率均较适中的提取条件。综合上述桑黄黄酮、糖苷酶抑制率的优化提取条件以及提取残渣中多糖的提取率,桑黄中降血糖成分较优的提取条件定为乙醇浓度60%,提取温度80℃,提取时间3h,料液比1∶30。在该条件下黄酮的提取率为86.4mg/g。该提取物对糖苷酶抑制率达99%。在该条件下,又对提取次数进行试验,结果如表2,提取次数以2次为宜。桑黄提取物对α-葡萄糖苷酶抑制的IC50约为25mg/L。
表2提取次数对黄酮含量和α-葡萄糖苷酶抑制率的影响
(2)多糖成分的提取
设立了四因素三水平的正交试验,称取桑黄适量,按表中的条件进行提取,提取结束后,进行离心,量取上清液体积并测定多糖含量,测定方法同前,试验结果如下表3所示:
表3
从试验结果表3可知,最优组合为温度100℃,提取时间2h料液比为1∶40,在此条件下的多糖的提取率为4.23%。把上一步提取α-葡萄糖苷酶抑制成分的残渣在此优化条件下进行提取。
与现有技术相比,本发明的有益效果是:
(1)本发明得到的具有α-葡萄糖苷酶抑制活性的提取物具有强烈的抑制活性,其抑制IC50可低至10μg/mL,远远低于降血糖药物阿卡波糖(165μg/mL);
(2)本发明既可较全面的利用了桑黄中的有效成分,提取了两类降血糖物质,又能从两个途径发挥降血糖作用,因此具有更强的降血糖效果;
(3)本发明提供的两步联合提取法具有较高的提取率,且方法工艺简单,安全环保,成本低廉,可操作性强,易于实际生产应用;
(4)本发明得到的提取物具有较好的降血糖效果,可以作为药品、保健品等作为辅助降血糖产品,提高了桑黄的附加值,为桑黄的利用提供了一条新途径,具有较好的应用前景。
说明书附图
图1是本发明从桑黄中降糖物质联合提取的工艺流程图。
图2是本发明糖尿病小鼠在喂养期间血糖的变化,
图中桑黄Ⅰ是指桑黄醇提取物,桑黄Ⅱ是指桑黄水提取物,桑黄Ⅲ是指桑黄醇提取物+桑黄水提取物。
图3是本发明糖尿病小鼠的糖耐量,图中桑黄Ⅰ是指桑黄醇提取物,桑黄Ⅱ是指桑黄水提取物,桑黄Ⅲ是指桑黄醇提取物+桑黄水提取物。
具体实施方式
下面结合实施例,更具体地说明本发明的内容。应当理解,本发明的实施并不局限于下面的实施例,对本发明所做的任何形式上的变通和/或改变都将落入本发明保护范围。
在本发明中,若非特指,所有的份、百分比均为重量单位,所有的设备和原料等均可从市场购得或是本行业常用的。若无特别指明,实施例采用的方法为本领域通用技术。
本发明的实施例中:
一种桑黄中降血糖物质的联合提取方法,包括如下步骤:
(1)桑黄子实体,干燥、粉碎并过筛,备用;
(2)按照固液比为1:10~1:30,在桑黄子实体粉末中加入乙醇,在温度60~80℃下提取1~3h,重复提取数次,冷却,离心,取上清液1备用;
(3)上清液1先用微孔滤膜过滤得到微滤液1,再利用截留分子量为10KD的超滤膜在压力为0.15~0.25Mpa、温度为25~40℃的工作条件下对微滤液1进行超滤,得到超滤液1,超滤液1进行减压浓缩,回收其中的乙醇,浓缩液1最后经真空冷冻干燥即得到桑黄提取物1,
(4)按照固液比1:20~1:40,在步骤(2)提取离心后剩余的残渣中加入水,在温度为90~100℃下提取1.5~2h,冷却,离心,取上清液2备用;
(5)上清液2先用微孔滤膜包过滤得到微滤液2,再利用截留分子量为1KD的复合超滤膜包在压力为0.1~0.15Mpa、温度为25~40℃的工作条件下对微滤液2进行超滤,取截留液,减压浓缩后再经真空冷冻干燥,得到桑黄提取物2。
需要注意的是,实施例1仅仅是对本发明最优实施例做出了说明。在本发明主要工艺参数范围内,本发明与现有技术相比均可实现比较好的提取率和α-葡萄糖苷酶抑制活性,实施例中不再赘述。
实施例1
(1)桑黄醇提取
桑黄子实体烘干(50℃),磨碎,过40目筛,称取100g粉末,加入60%浓度乙醇溶液3000ml,控制温度为70℃,提取3h后,趁热把提取液移入离心杯中,在5000rpm下离心20min,收集上清液和渣,在渣中继续加入60%浓度乙醇,在相同条件下提取1次,离心后收集上清液和渣,两次上清液合并。渣用于下一步的水提。上清液用超滤杯先过0.45μm的聚醚砜膜进行微滤,得到的滤液再用10KD的聚醚砜超滤膜超滤,工作压力为0.2Mpa,温度为室温。微滤和超滤时,膜需要定期更换,以防止过滤速度太慢。超滤液进行减压浓缩,回收乙醇,浓缩液进行真空冻干,得到桑黄醇提取物(即α-葡萄糖苷酶抑制成分提取物)20g。
经检测,黄酮的提取率达到159.3mg/g原料。
该桑黄醇提取物用二甲基亚砜配制成浓度为10g/L溶液,并用pH6.8的磷酸缓冲溶液梯度稀释,测定各稀释液的抑制率,得到IC50为10μg/mL。
(2)桑黄水提取
取步骤(1)醇提后剩下的渣,干燥后约80g,加入水300ml,温度控制在100℃,提取2h,趁热离心,离心条件同前,收集上清液。上清液先通过0.45μm的混合纤维素微孔滤膜包微滤,滤液用1KD的复合超滤膜包(聚醚砜+聚酰胺)截留,取截留液,截留液为初始溶液体积的三分之一时,停止超滤,截留液减压浓缩,真空冷冻干燥得冻干粉-即桑黄水提取物10g,冻干粉中多糖含量为18.3%。
对比例1
本发明中的桑黄粉末,按照李善姬等报道的文献中的提取方法进行提取,即乙醇浓度60%,料液比1∶50(g/mL),超声功率80W,提取时间40min,结果黄酮的提取率为23.5mg/g原料,所得的提取物对α-葡萄糖苷酶抑制的IC50为0.642g/L。
对比例2
对本发明中的桑黄粉末直接提取其中的多糖,提取条件按照前面最优化的条件即提取温度100℃,提取时间2h,料液比1∶40,提取1次,结果多糖的提取率为41mg/g原料。
实验例部分
实验例1不同乙醇浓度提取物的α-葡萄糖苷酶抑制活性对比
桑黄子实体粉末分别用0、20、40、60、80、100%浓度乙醇提取(按固液比1∶20在60℃下提取1h),经离心(5000rpm,20min),收集上清液,减压浓缩,真空冷冻干燥得到提取物。配制各提取物溶液10g/L,并用pH值6.8的磷酸缓冲溶液梯度稀释,测定各稀释液的α-葡萄糖苷酶抑制率,计算IC50,得到各浓度乙醇提取的IC50分别为315、80、52、21、35μg/mL。
实验例2桑黄降血糖提取物的体内降血糖作用
实验材料:桑黄醇提取物+水提物
实验设计:ICR小鼠(6~8周)100只,重量为18~22g,饲养在一个温度可控(24±2℃)、相对湿度为50±5%、具有12h光照周期的房间里,基础饲料喂养,可自由觅食。适应性喂养一周后,随机挑出15只鼠作为正常组,其它鼠高脂饲料喂养4周,四周后,禁食15~16h,从尾静脉注射链脲霉素STZ(30mg/kg),第二天再注射一次。继续高脂喂养10天,测定尾静脉的血糖值,挑出血糖值高于11.1mmol/L或空腹血糖值高于8mmol/L的鼠用于下一步的实验。实验鼠随机分成6组,每组10只,即模型组、阳性对照组和四个药物组。正常组和模型组每天灌胃0.1mL/10g BW剂量的生理盐水;阳性对照组每天灌胃100mg/kg BW的二甲双胍;四个药物组分别灌胃50、100mg/kg BW的醇提物、50mg/kg BW的水提物和50mg/kg BW的醇提物+50mg/kg BW的水提物。所有的组连续灌胃8周,体重和空腹血糖值每10天测量一次。口服糖耐量的测定在第7周禁食1夜后进行。糖尿病小鼠在喂养期间和喂养结束时的体重、血糖变化以及糖耐量如附表和附图所示。从附表和附图可以看出,在桑黄药物喂养期间,糖尿病小鼠的体重逐渐增加,血糖值缓慢下降。至喂养结束,桑黄组的体重明显低于模型组(P<0.05),血糖值明显低于模型组(P<0.05),醇提物和水提物有一定的协同作用,1∶1混合物(醇提物+水提物)明显高于它们单独作用组,最终1∶1混合物喂养组的血糖值降至最低。
表4小鼠喂养期间体重的变化(g)
组别 | 0d | 10d | 20d | 30d | 40d | 50d | 56d |
正常对照组 | 25.2±1.1 | 26.4±1 | 28.7±1.1 | 31.1±1.5 | 33±1.5 | 34.9±1.2 | 35.9±1.5 |
模型对照组 | 28.2±1 | 29.7±1.2 | 32.4±1.3 | 35±1.4 | 38.3±1.5 | 40.4±1.6 | 42.1±2.2 |
二甲双胍 | 28.4±1.1 | 29.6±1.2 | 32.3±1.6 | 34.1±1.7 | 36.2±1.7 | 38.4±1.9 | 39.8±2.3 |
醇提物低 | 28.5±1.4 | 30.1±1.6 | 32.7±1.8 | 34.8±2 | 37±1.9 | 39.4±2.1 | 40.7±2.5 |
醇提物高 | 28±1.9 | 29.2±2.1 | 32±2.6 | 34.2±2.2 | 36.1±2 | 38.1±2.2 | 39.4±2.1 |
水提物 | 27.5±1.9 | 28.3±2 | 31.1±2.1 | 33.5±2.3 | 35.7±2.7 | 37.7±3 | 39.7±2.9 |
1∶1混合物 | 28.6±1.8 | 29.8±2 | 32.7±2.2 | 34.7±2.3 | 37.1±2.2 | 39.2±2.5 | 40.8±2.7 |
表5糖尿病小鼠喂养结束时的体重和血糖值
实验例3桑黄抑制α-葡萄糖苷酶成分鉴定
抑制α-葡萄糖苷酶提取物(桑黄醇提取物)通过Waters UPLC-Synapt G2液质联用分析,色谱柱:ACQUITY UPLC BEH C18(100mm×2.1mm,1.7μm),流动相:0.1%甲酸水溶液(B)和乙腈(A),进样室温度:10℃,柱温:35℃,洗脱梯度(0~2min,95%~80%A;2~5min,80%~70%A;5~8min,70%~65%A;8~10min,65%~35%A;10~20min,35%~2%A;20~23min,2%~2%A;23~23.1min,2%~95%A;23.1~26min,95%A),流速:0.3mL/min,进样量:1μL。
质谱条件:电喷雾离子源,正,负离子模式下扫描,数据采用一级母离子全扫描和数据依赖性前三强二级子离子扫描(Full Scan MS/dd-MS2),质谱扫描范围:120~1 500m/z;一级分辨率为35 000,二级分辨率为17500;隔离窗口为2.0m/z:喷雾电压(+)为3.5kV,喷雾电压(-)为3.0kV;鞘气压力为30arb;辅助气压力为10arb;毛细管温度为320℃;S-lens为50;碰撞能量为30,40,60%。
通过对照品,参考文献和数据库,对桑黄的抑制糖苷酶提取物进行靶向鉴定了多酚、黄酮和萜类成分,部分成分的保留时间,准分子离子峰的质荷比,碎片离子、化学名称等数据,如表6所示,其中原儿茶酸、咖啡酸、7,8-二羟基-4-甲基香豆素、橘霉素为多酚类物质,山柰酚和桑色素D为黄酮类物质,咖啡豆醇、桦木酸和熊果酸为萜类物质。另外提取物中还大量存在着吡喃酮结构的酚类物质,即Phelligridin J、Phelligridin D、Phelligrindin G、phelligridimer A、Interfungin B、Phellibaumin E、Inoscavin A、inoscavin C、Phellifuropyranone、Phellinignincisterol A、Phellilins C等,这些物质在其它的桑黄中也已被鉴定出。
表6桑黄中抑制α-葡萄糖苷酶提取物LC-MS鉴定结果
参考文献:
[1]齐欣,张峻,陈颖等,六种不同树种桑黄有效成分的比较.食品科学,2010,31(6):199-201.
[2]李善姬,梁承武,崔承弼,等,野生桑黄总黄酮提取工艺及其抗氧化作用的研究,食品研究与开发,2015,36(20):32-35.
[3]冯子旺,俞力超,李峰,等,正交试验优选桑黄多酚超声提取工艺.中国药房,2012,23(3):221-222.
[4]陈晓平,于翠翠.响应面法优化微波辅助乙醇提取桑黄黄酮工艺的研究,食品与发酵科技,49,(4):31-36.
[5]回晶,李其久,边媛媛,等,桑黄总黄酮超声提取工艺及其生物活性研究.食品科学,2010,31(24):195-198.
[6]曾鹏,黄世荣,刘明明等,应用响应面法优化超声波辅助热水提取桑黄多糖的工艺条件.蚕业科学,2015,41(5):0928-0933.
[7]严红实,金乾坤,李鹏飞等,桑黄多糖的提取及抗氧化性研究,食品科技,2015,38(6):201-205.
[8]张作法.桑黄中活性物质的高效制备方法:中国,CN201910619234.7[P].2020-03-17.
[9]田东升,孟箫.一种桑黄中活性成分的高效制备方法:中国,CN201910619234.7[P].2019-11-05.
[10]Zheng,M.,Lu,S.,&Xing,J.(2021).Enhanced antioxidant,anti-inflammatory andα-glucosidase inhibitory activities of citrus hesperidin byacid-catalyzed hydrolysis.Food Chemistry,336,127539.
Claims (10)
1.一种桑黄中降血糖物质的联合提取方法,其特征在于,所述的联合提取方法包括如下步骤:
(1)桑黄子实体,干燥、粉碎并过筛,备用;
(2)按照固液比为1:10~1:30,在桑黄子实体粉末中加入乙醇,在温度60~80℃下提取1~3h,重复提取数次,冷却,离心,取上清液1备用;
(3)上清液1先用微孔滤膜过滤得到微滤液1,再利用截留分子量为10KD的超滤膜在压力为0.15~0.25Mpa、温度为25~40℃的工作条件下对微滤液1进行超滤,得到超滤液1,超滤液1进行减压浓缩,回收其中的乙醇,浓缩液1最后经真空冷冻干燥即得到桑黄提取物1,
(4)按照固液比1:20~1:40,在步骤(2)提取离心后剩余的残渣中加入水,在温度为90~100℃下提取1.5~2h,冷却,离心,取上清液2备用;
(5)上清液2先用微孔滤膜包过滤得到微滤液2,再利用截留分子量为1KD的复合超滤膜包在压力为0.1~0.15Mpa、温度为25~40℃的工作条件下对微滤液2进行超滤,取截留液,减压浓缩后再经真空冷冻干燥,得到桑黄提取物2。
2.根据权利要求1所述的一种桑黄中降血糖物质的联合提取方法,其特征在于,所述的步骤(1)中桑黄子实体在50~70℃下干燥,粉碎后过30~60目筛。
3.根据权利要求1所述的一种桑黄中降血糖物质的联合提取方法,其特征在于,所述的步骤(2)中乙醇浓度为40~80%。
4.根据权利要求1所述的一种桑黄中降血糖物质的联合提取方法,其特征在于,所述的步骤(3)中微孔滤膜采用聚醚砜微孔滤膜;超滤膜采用聚醚砜超滤膜。
5.根据权利要求1所述的一种桑黄中降血糖物质的联合提取方法,其特征在于,所述的步骤(5)中微孔滤膜包采用混合纤维素微孔滤膜包;复合超滤膜包采用聚醚砜-聚酰胺复合超滤膜包。
6.根据权利要求1所述的一种桑黄中降血糖物质的联合提取方法,其特征在于,所述的步骤(5)中超滤:回流液流回原液中,当截取液体积减少至原液初始体积的30%-35%,透过液速度明显降低时,停止超滤。
7.桑黄提取物在制备α-葡萄糖苷酶抑制剂上的应用。
8.根据权利要求7所述的应用,其特征在于,所述的桑黄提取物按下述方法提取:
按照固液比为1:10~1:30,在桑黄子实体粉末中加入乙醇,在温度60~80℃下提取1~3h,重复提取数次,冷却,离心,取上清液1备用;
上清液1先用微孔滤膜过滤得到微滤液1,再利用截留分子量为10KD的超滤膜在压力为0.15~0.25Mpa、温度为25~40℃的工作条件下对微滤液进行超滤,得到超滤液1,超滤液1进行减压浓缩,回收其中的乙醇,浓缩液1最后经真空冷冻干燥即得到桑黄提取物1。
9.根据权利要求8所述的应用,其特征在于,所述的微孔滤膜采用聚醚砜微孔滤膜;超滤膜采用聚醚砜超滤膜。
10.根据权利要求7所述的应用,其特征在于,所述的桑黄提取物为醇提取物,其组成包括下列成分:
原儿茶酸、咖啡酸、二氢山柰酚、7,8-二羟基-4-甲基香豆素、山柰酚、橘霉素、桑色素D、咖啡豆醇、脱氢枞酸、桦木酸和熊果酸。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113893269A (zh) * | 2021-10-05 | 2022-01-07 | 浙江省农业科学院 | 具有降血压功效的芹菜籽提取液的制备及纯化方法 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101474211A (zh) * | 2008-10-27 | 2009-07-08 | 上海市农业科学院 | 一种桑黄提取物及其制备方法 |
CN102106875A (zh) * | 2010-12-06 | 2011-06-29 | 浙江工业大学 | 一种水溶性α-葡萄糖苷酶抑制剂的提取方法 |
KR20150131671A (ko) * | 2014-05-16 | 2015-11-25 | 박준덕 | 금사린테우산을 주성분으로 하는 주사제용 항암제 조성물 및 그 제조방법 |
CN111096983A (zh) * | 2020-01-17 | 2020-05-05 | 浙江工业大学 | 具有降血糖活性的桑黄酚类提取物及其制备与应用 |
-
2020
- 2020-12-17 CN CN202011494218.9A patent/CN112546077A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101474211A (zh) * | 2008-10-27 | 2009-07-08 | 上海市农业科学院 | 一种桑黄提取物及其制备方法 |
CN102106875A (zh) * | 2010-12-06 | 2011-06-29 | 浙江工业大学 | 一种水溶性α-葡萄糖苷酶抑制剂的提取方法 |
KR20150131671A (ko) * | 2014-05-16 | 2015-11-25 | 박준덕 | 금사린테우산을 주성분으로 하는 주사제용 항암제 조성물 및 그 제조방법 |
CN111096983A (zh) * | 2020-01-17 | 2020-05-05 | 浙江工业大学 | 具有降血糖活性的桑黄酚类提取物及其制备与应用 |
Non-Patent Citations (1)
Title |
---|
徐秋燕等: "植物中黄酮类化合物的提取方法", 《纺织科技进展》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113893269A (zh) * | 2021-10-05 | 2022-01-07 | 浙江省农业科学院 | 具有降血压功效的芹菜籽提取液的制备及纯化方法 |
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