CN115006448A - 一种高含量北豆根生物总碱及其制备方法 - Google Patents
一种高含量北豆根生物总碱及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种高含量北豆根生物总碱及其制备方法,涉及天然药物的提取、分离技术领域,包括:将北豆根经一定比例的乙醇溶剂提取后,提取液经有机溶剂萃取得生物碱粗品,生物碱粗品经阳离子交换色谱柱聚类得到生物碱组分,即为北豆根生物总碱。本发明的有益效果是利用北豆根提取液经不同有机溶剂萃取后经阳离子交换色谱柱聚类制备后即可除去非生物碱类成分,富集生物碱类组分,生物碱的损失率低,含量高;本发明技术重现性好,适应性广,快速高效,易于工业化生产。
Description
技术领域
本发明涉及天然药物的提取、分离技术领域,具体涉及一种高含量北豆根生物总碱的制备方法。
背景技术
北豆根为防己科植物蝙蝠葛(Menispermum dauricum)的干燥根茎,在我国主要分布于东北、华北、华中、华东、陕西等地。2015年版中国药典记载北豆根气微、味苦、寒,归肺、胃、大肠经,具有清热解毒、祛风止痛之功效,可用于治疗咽喉肿痛、肠炎痢疾、风湿痹病等症。此外临床上还多应用于肺热咳嗽、扁桃体炎、慢性支气管炎等疾病的治疗。研究表明北豆根中生物碱类成分为其特征成分和生物活性成分,不仅具有基于传统功效的抑菌、抗炎等药理活性,而且具有保护心脑血管系统、抗肿瘤、抗抑郁、抗阿尔茨海默症等生物活性,其总生物碱含量为1.7%~2.5%,生物碱类型主要为双苄基四氢异喹啉型生物碱、氧化异阿朴啡型生物碱、吗啡烷型生物碱、阿朴啡型生物碱、原小檗碱型生物碱、异喹啉和异吲哚型生物碱。
目前以北豆根总生物碱为主要成分的批件中成药包括北豆根咀嚼片、北豆根滴丸、北豆根分散片、北豆根片、北豆根胶囊及复方北豆根氨酚那敏片等,均以提取物为原料,质量标准低,产品品质不高。目前,以中药组分为原料的中药新药的研制具有疗效高、剂量小的优势,而市场常规供应的北豆根总碱提取物的总碱含量10%~20%左右,含量较低。因此迫切需要获取高含量的北豆根生物总碱,为中药组分新药研究提供高品质原料和制备工艺。
目前北豆根总碱提取工艺的常规方法主要为酸提碱沉后有机溶剂萃取,该方法富集总碱的同时,有大量酸碱性杂质也一同富集,导致总碱含量不高、专属性差等问题。少数报道采用大孔吸附树脂、阴离子交换树脂进一步富集纯化。其中大孔吸附树脂分离工艺简单、成本低、易于产业化,但该方法存在载样量低、制备效率低等缺点。如中国专利CN111317754公开了一种北豆根总碱的制备方法及应用,该专利采用的是阴离子交换色谱柱,其原理是将酸性成分吸附,而生物碱等其他成分直接流穿下来,故而导致精制目标馏分回收率高,但其生物碱含量较低。
因此,需要发展高效高选择性富集北豆根总碱的制备工艺,获取高含量的总生物碱,为北豆根生物碱组分新药研发奠定重要基础。
发明内容
本发明的目的在于至少解决现有技术中存在的技术问题之一,提供一种高含量北豆根生物总碱的制备方法。具体方法为北豆根药材经一定比例的乙醇溶剂提取后,提取液经有机溶剂萃取得生物碱粗品,生物碱粗品经阳离子交换色谱柱制备得到生物碱组分。
本发明的技术解决方案如下:
一种高含量北豆根生物总碱的制备方法,包括以下步骤:
S1、北豆根提取液的获取:称取北豆根饮片,加入体积分数为50~95%的乙醇溶液浸泡,加热回流提取,过滤取滤液,滤液经浓缩后得到提取液;
S2、生物碱粗品的萃取:在步骤S1所得提取液中加入酸水溶液,搅拌、超声溶解,待提取液显强酸性后加入有机溶剂1萃取,得到有机溶剂1层和酸水层;在酸水层中加入碱性溶液,至显强碱性后再加入有机溶剂2萃取,得到有机溶剂 2层和碱水层,将有机溶剂2层经浓缩后得到生物碱粗品;
S3、制备上样液的酸化:将步骤S2所得生物碱粗品加入体积分数为50~90%的甲醇水溶液中,待大部分样品溶解后加酸进行酸化,并将酸化后的样品进行超声、离心至样品全部溶解,经0.20~0.50μm滤膜后得制备上样液;
S4、生物碱组分的聚类:离子交换色谱柱加入酸和体积分数为30~90%的甲醇水溶液,然后取步骤S3所配置的制备上样液上样,待上样结束后加入酸和体积分数为50~90%的甲醇水溶液,洗脱非生物碱成分,然后加入盐及体积分数为的50~90%的甲醇水溶液,洗脱生物碱成分,洗脱的生物碱成分经浓缩、冻干,即得目标生物碱组分。
优选地,步骤S1中,北豆根饮片与乙醇溶液的料液比为1g:8~10ml,浸泡 1~24h,在50~85℃下回流提取0.5~3小时,然后重复提取1~2次,滤液合并。
优选地,步骤S1中,乙醇溶液的体积分数为70%,在60℃下回流提取2小时。
优选地,步骤S2中,酸水溶液为质量分数为70%的稀硫酸溶液,调步骤S1 所得提取液PH至2~3;碱性溶液为氢氧化钠溶液和/或氨水溶液,调酸水层PH 至9~10;有机溶剂1为乙酸乙酯,有机溶剂2为二氯甲烷和正丁醇;萃取次数为2~4次。
优选地,步骤S3中,酸与甲醇水溶液的体积比为0.1%~5%,制备上样液的浓度为50~200mg/mL。
优选地,步骤S4具体包括:
离子交换色谱柱加入酸和2~3倍柱体积的30~90%甲醇水溶液,酸与甲醇水溶液的体积比为0.1~5.0%;然后取步骤S3所配置的制备上样液上样,载样量为 1.0~3.0%;待上样结束后加入酸和2~5倍柱体积的50~90%甲醇水溶液,酸与甲醇水溶液的体积比为0.1~5.0%,洗脱非生物碱成分;然后加入盐和8~15倍柱体积的50~90%甲醇水溶液,盐的摩尔浓度为80~300mM/L,洗脱生物碱成分。
优选地,步骤S4中,酸为甲酸,盐为甲酸铵。
优选地,步骤S4中,离子交换色谱柱填料为HSCX,填料粒径为40μm,色谱柱内径大小为10~100mm,柱长为250mm,样品载样量为1.0~5.0%。
优选地,步骤S4中,最后加入2~3倍柱体积的50%~90%(V/V)甲醇水溶液(需加一定摩尔浓度的盐,其浓度为80~300mM/L)冲柱,待制备结束后加入 3倍柱体积的90%甲醇水溶液(加一定摩尔浓度的盐,其浓度为10mM/L)保存色谱柱。
一种高含量北豆根生物总碱,采用上述的方法得到。
本发明至少具有以下有益效果之一:
1、本发明提供了一种获取高含量的北豆根生物总碱的新方法,本发明先是加入50~95%乙醇溶液对北豆根进行提取,得到提取液;然后对提取液进行萃取、浓缩,得到生物碱粗品;再往生物碱粗品加入50~90%甲醇水溶液和酸,由于北豆根植物中含有大量的异喹啉类生物碱,且该类生物碱成分极性较小,故本发明通过加适量的酸可提高其溶解度,即提高样品溶解度,增加样品浓度,减少上样体积;此外,加酸还可使弱碱性生物碱成分充分电离,从而与阳离子交换色谱柱有足够强的结合力,进而增加制备载样量。最后,将样品加入阳离子交换色谱柱,通过阳离子交换色谱柱将非生物碱成分去除,达到富集生物碱成分的目的,阳离子交换色谱柱富集生物碱组分的原理是将中性的小极性和碱性成分保留在色谱柱上,酸性体积分数为50~90%甲醇溶剂可将中性成分洗脱,而含盐的体积分数为50~90%的甲醇才可将生物碱成分洗脱,由于北豆根含大量的生物碱成分,且该类成分主要为异喹啉类生物碱,其极性较小,碱性适中,因此,本发明采用阳离子交换色谱柱可富集纯度较高的生物碱成分,且生物碱成分损失较少,从而得到高含量北豆根生物总碱。本发明的方法中饮片仅需经提取、萃取、制备上样液酸化和阳离子交换色谱柱聚类等步骤,载样量可达3%,方法操作简便、普适性强、易于产业化。
2、本发明制备得到的北豆根生物总碱的含量高达92%,比富集前的粗碱的总碱含量提高了30%,相比北豆根提取物总碱含量提高了60%以上。因此,在北豆根的总生物碱制备上具有明显优势。
附图说明
图1为本发明的提取工艺流程图;
图2为实施例1中北豆根生物碱组分制备色谱图;
图3为实施例1中北豆根生物碱粗品液相色谱图;
图4为实施例1中北豆根富集后生物碱组分液相色谱图;
图5为实施例1中北豆根生物碱萃取物、北豆根F2馏分的总生物碱含量对比图,其中,峰1:附子亭;峰2:附子亭异构体;峰3:蝙蝠葛苏林碱;峰4:蝙蝠葛碱。
具体实施方式
下面用具体实施例对本发明做进一步详细说明,但本发明不仅局限于以下具体实施例。
实施例1:
如图1所示,本实施例提供一种高含量北豆根生物总碱的制备方法,包括以下步骤:
S1、北豆根提取液的获取:取100g北豆根饮片,按料液比1:10加入1000 mL的70%(V/V)乙醇水浸泡24h后于60℃加热回流提取2h,共提取3次(重复以上操作再提2次),提取后过滤,合并3次滤液后浓缩至1L,得北豆根提取液。其中北豆根提取率为18.76%。
S2、北豆根生物碱粗品的萃取:北豆根提取液中加适量70%稀硫酸溶液,并超声、溶解、搅拌调pH至2~3,加入与提取液(调pH后的提取液)相同体积的乙酸乙酯重复萃取3次,得乙酸乙酯层和酸水层,酸水层中加入适量氨水调pH 至9~10,加入与酸水层(调pH后的酸水层)相同积体的正丁醇重复萃取3次,得碱水层和正丁醇层。经检测分析,正丁醇层主要含有生物碱类成分,经浓缩后可得北豆根生物碱粗品7.28g,其占提取饮片总质量的7.28%;
S3、北豆根制备上样液的酸化:取一定量的北豆根生物碱粗品,加入适量 70%(V/V)甲醇水和甲酸(甲酸与70%(V/V)甲醇水溶液的体积比为0.1%)溶液,经超声、震荡后过0.45μm滤膜,得北豆根制备上样液,其浓度为100mg/mL;
S4、北豆根生物碱组分的聚类:阳离子交换色谱柱(填料为HSCX,粒径为 40μm,柱规格为10×250mm,填料质量为12.0g)经2倍柱体积的50%(V/V) 甲醇水(需加甲酸,甲酸与50%(V/V)甲醇水溶液体积比为0.1%,V/V)平衡后,取上样样品载样量(上样样品中固体的总质量与填料质量比)为3.0%的实验步骤S3配置的北豆根上样液至阳离子交换色谱柱中,先经2倍柱体积的50% (V/V)甲醇水溶液(需加甲酸,甲酸与50%(V/V)甲醇水溶液体积比为0.1%, V/V)洗脱,再经8倍柱体积50%(V/V)甲醇水溶液(需加甲酸铵,其摩尔浓度为200mM/L)洗脱,最后加3倍柱体积50%(V/V)甲醇水溶液(需加甲酸铵,其摩尔浓度为200mM/L)冲柱,其中50%(V/V)甲醇水溶液(盐为甲酸铵,其摩尔浓度为200mM/L)洗脱液即为北豆根生物总碱馏分,其经浓缩后可得目标生物碱组分。
结果分析:
(一)对富集前的生物碱粗品和富集后的北豆根生物碱组分进行同条件液相分析,结果如图2~4所示,其中,图2为北豆根生物碱组分制备色谱图,图3 为北豆根生物碱粗品液相色谱图,图4为北豆根富集后生物碱组分液相色谱图。在图2中北豆根生物碱组分制备色谱图中,目标北豆根生物碱组分为182.6mg,占总上样质量的50.7%,经检测分析,该3%载样量下生物碱穿透仅为7.86%,总碱损失率在10%以内。
将图3中北豆根生物碱粗品液相色谱图与图4中北豆根生物碱组分液相色谱图进行对比可知,北豆根生物碱粗品经过富集后生物碱组分没有丢失,全保留,由此说明,本发明的富集方法不会导致生物碱组分的丢失。
(二)实施例1中北豆根生物碱提取物、萃取物、馏分的总生物碱含量对比实验
以北豆根生物碱提取物、萃取物、馏分的总生物碱含量为研究对象,采用确定的分析方法,以蝙蝠葛苏林碱、蝙蝠葛碱为对照,测定实施例1中北豆根生物碱提取物(即步骤S1得到的北豆根提取液)、萃取物(即步骤S2得到的北豆根生物碱粗品)、馏分(即步骤S4得到的北豆根生物总碱馏分)的总生物碱含量。
实验方法
1)供试品溶液制备
取北豆根生物碱提取物(2.10mg)、萃取物(1.66mg)、F2馏分(1.88mg),置5mL容量瓶中,加入1%甲酸甲醇,定容至刻度,超声处理(功率140W,频率42kHz)30分钟,取出,放冷,滤过,取续滤液,即得。
2)对照品溶液制备
蝙蝠葛苏林碱对照品溶液(238.1400μg/mL)
蝙蝠葛碱对照品溶液(259.0760μg/mL)。
3)液相方法
仪器:Waters e2695(WHPLC-10)
色谱柱:C18HCE(4.6×100mm,3μm,SN:C18HCE-4)
流动相:A:0.1%甲酸乙腈B:0.1%甲酸水
流速:1mL/min
进样体积:10μL
波长:280nm
柱温:30℃
洗脱条件:
时间(分钟) | A% | B% |
0 | 7 | 93 |
2 | 7 | 93 |
7 | 15 | 85 |
12 | 90 | 10 |
实验结果见表1和图5,其中,表1为北豆根生物碱提取物、萃取物、馏分的总生物碱含量对比,图5为北豆根生物碱萃取物、北豆根F2馏分的总生物碱含量对比图。
表1北豆根生物碱提取物、萃取物、馏分的总生物碱含量对比
备注:总生物碱以蝙蝠葛苏林碱对照品的峰面积为对照,峰1~峰4峰面积总和计算。由于是默认峰1、2与峰3校正因子为1计算。
由图5和表1可知,总生物碱以蝙蝠葛苏林碱对照品的峰面积为对照,北豆根F2馏分中总生物碱含量达到92.82%,富集前的北豆根萃取物(粗碱)总碱含量61.57%,北豆根提取物总碱含量29.10%,其中目标生物碱组分经过阳离子交换色谱柱HSCX富集后,相比富集前的萃取物总碱含量提高了31.25%,相比北豆根提取物总碱含量提高了63.72%,因此,本发明中的北豆根异喹啉生物碱组分富集方法可以有效除杂并提高总碱含量,大大提高纯度。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (10)
1.一种高含量北豆根生物总碱的制备方法,其特征在于,包括以下步骤:
S1、北豆根提取液的获取:称取北豆根饮片,加入体积分数为50~95%的乙醇溶液浸泡,加热回流提取,过滤取滤液,滤液经浓缩后得到提取液;
S2、生物碱粗品的萃取:在步骤S1所得提取液中加入酸水溶液,搅拌、超声溶解,待提取液显强酸性后加入有机溶剂1萃取,得到有机溶剂1层和酸水层;在酸水层中加入碱性溶液,至显强碱性后再加入有机溶剂2萃取,得到有机溶剂2层和碱水层,将有机溶剂2层经浓缩后得到生物碱粗品;
S3、制备上样液的酸化:将步骤S2所得生物碱粗品加入体积分数为50~90%的甲醇水溶液中,待大部分生物碱粗品溶解后加酸进行酸化,并将酸化后的生物碱粗品进行超声、离心至样品全部溶解,经0.20~0.50μm滤膜后得制备上样液;
S4、生物碱组分的聚类:离子交换色谱柱加入酸和体积分数为30~90%的甲醇水溶液,然后取步骤S3所配置的制备上样液上样,待上样结束后继续加入酸和体积分数为50~90%的甲醇水溶液,洗脱非生物碱成分,然后加入盐及体积分数为的50~90%的甲醇水溶液,洗脱生物碱成分,洗脱的生物碱成分经浓缩、冻干,即得目标生物碱组分。
2.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S1中,北豆根饮片与乙醇溶液的料液比为1g:8~10ml,浸泡1~24h,在50~85℃下回流提取0.5~3小时,然后重复提取1~2次,滤液合并。
3.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S1中,乙醇溶液的体积分数为70%,在60℃下回流提取2小时。
4.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S2中,酸水溶液为质量分数70%的稀硫酸溶液,调步骤S1所得提取液PH至2~3;碱性溶液为氢氧化钠溶液和/或氨水溶液,调酸水层PH至9~10。
5.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S2中,有机溶剂1为乙酸乙酯,有机溶剂2为二氯甲烷和正丁醇,采用有机溶剂1和有机溶剂2各萃取2~4次。
6.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S3中,酸与甲醇水溶液的体积比为0.1~5%,制备上样液的浓度为50~200mg/mL。
7.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S4具体包括:
离子交换色谱柱加入酸和2~3倍柱体积的30~90%甲醇水溶液,酸与甲醇水溶液的体积比为0.1~5.0%;然后取步骤S3所配置的制备上样液上样,载样量为1.0~3.0%;待上样结束后加入酸和2~5倍柱体积的50~90%甲醇水溶液,酸与甲醇水溶液的体积比为0.1~5.0%,洗脱非生物碱成分;然后加入盐和8~15倍柱体积的50~90%甲醇水溶液,盐的摩尔浓度为80~300mM/L,洗脱生物碱成分。
8.根据权利要求7所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S4中,酸为甲酸,盐为甲酸铵。
9.根据权利要求1所述的一种高含量北豆根生物总碱的制备方法,其特征在于,步骤S4中,离子交换色谱柱填料为HSCX,填料粒径为40μm,色谱柱内径大小为10~100mm,柱长为250mm,样品载样量为1.0~5.0%。
10.一种高含量北豆根生物总碱,其特征在于,采用权利要求1~9任一所述的方法得到。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995001984A1 (en) * | 1993-07-06 | 1995-01-19 | Polis A.G. | Process for the extraction and purification of alkaloids |
CN101647828A (zh) * | 2008-08-13 | 2010-02-17 | 代龙 | 一种用离子交换树脂分离中药总生物碱的方法 |
CN101962360A (zh) * | 2010-10-26 | 2011-02-02 | 南京泽朗农业发展有限公司 | 一种从蝙蝠葛中提取蝙蝠葛碱的方法 |
CN102001997A (zh) * | 2010-10-26 | 2011-04-06 | 南京泽朗农业发展有限公司 | 一种蝙蝠葛碱的精制方法 |
CN111317754A (zh) * | 2018-12-13 | 2020-06-23 | 泰州医药城国科化物生物医药科技有限公司 | 一种北豆根总碱的制备方法及应用 |
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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995001984A1 (en) * | 1993-07-06 | 1995-01-19 | Polis A.G. | Process for the extraction and purification of alkaloids |
CN101647828A (zh) * | 2008-08-13 | 2010-02-17 | 代龙 | 一种用离子交换树脂分离中药总生物碱的方法 |
CN101962360A (zh) * | 2010-10-26 | 2011-02-02 | 南京泽朗农业发展有限公司 | 一种从蝙蝠葛中提取蝙蝠葛碱的方法 |
CN102001997A (zh) * | 2010-10-26 | 2011-04-06 | 南京泽朗农业发展有限公司 | 一种蝙蝠葛碱的精制方法 |
CN111317754A (zh) * | 2018-12-13 | 2020-06-23 | 泰州医药城国科化物生物医药科技有限公司 | 一种北豆根总碱的制备方法及应用 |
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