CN108969555B - 一种补肾、益精血、增强免疫力的淫羊藿中药组合物的制备方法 - Google Patents
一种补肾、益精血、增强免疫力的淫羊藿中药组合物的制备方法 Download PDFInfo
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Abstract
公开了一种补肾、益精血、增强免疫力的淫羊藿红景天中药组合物,以及雪莲提取装置,所述提取装置效率高,并且能够最大限度地减少提取过程中副产物的产生,从而使得到的提取物具有较高的药理活性。所述组合物具有提高免疫力、抗炎、抗疲劳、防辐射并且补肾、益精血的功效。
Description
技术领域
本发明涉及一种淫羊藿红景天中药组合物的制备方法,特别涉及一种提高免疫力、抗炎、抗疲劳、防辐射并且补肾、益精血的中药组合物的制备方法,所述中药组合物尤其是包含雪莲提取物、淫羊藿提取物和红景天提取物的组合物,即雪莲淫羊藿红景天中药组合物。
背景技术
雪莲是我国珍稀名贵的中药材,有着久远的要用历史以及丰富的民间用药基础,雪莲中含有的黄酮及其苷、苯丙素类、倍半萜内酯、香豆素、木脂素、有机酸、氨基酸、甾体、多糖等成分,具有抗风湿、镇痛、调节心血管***、抗癌、延缓衰老、防辐射等生物活性,有着很大的药用开发价值。在很多文献中记载了天山雪莲及其提取物的提高免疫力、抗炎、防辐射功效。《苏州大学》,高博,2001,“天山雪莲提取物的抗辐射损伤作用及机制研究”,指出天山雪莲花作为传统药物沿用已久,药理作用明确,具有很好的抗氧化、清除自由基的作用,该文通过整体动物实验、羟自由基清除实验、染色体畸变的Giemsa染色分析以及调控细胞周期的P21基因表达的RT-PCR分析,***地论证了天山雪莲花提取物的抗辐射损伤作用及其可能的作用机理。《中国辐射卫生》,2010年4期,郭娜等,“苞叶雪莲水提物对小鼠生殖细胞辐射防护作用的研究”,研究了苞叶雪莲水提物对小鼠生殖细胞的辐射防护作用。《中药药理与临床》,黄华等,《中药药理与临床》,2002,“雪莲花口服液的活血镇痛和抗炎作用研究”,研究了雪莲花口服液的提高免疫力以及活血、镇痛和抗炎作用。
淫羊藿的补肾和益精血作用和机理也得到了广泛认同和研究。在李梅编著的全国高等医药院校药学类第四轮规划教材《中医药学基础》(第3版[M].2016)中,明确指出淫羊藿具有补肾壮阳功效,其药理主要是增强免疫功能、提高性腺功能和延缓衰老、抗炎、抗过敏等。在国家食品药品监督管理总局执业药师资格认证中心组织编写的《中药学专业知识》(2015)也明确指出淫羊藿及其提取物(主要是黄酮类和苷类化合物)的温肾助阳作用。在张惊湖编著的《常用中药功效比较》(2013)明确指出了淫羊藿及其提取物的补肾、益精血作用。
红景天具有益气、活血、化淤的功效。《中国临床药理学与治疗学》,姜爱玲等,“红景天苷药理作用的研究进展”,2015,指出红景天苷是我国传统藏药红景天的主要活性成分之一,药用价值很高,该文从红景天苷对骨骼***、心血管***、神经***、肝脏的保护作用,对糖尿病肾病的治疗作用、抗炎症、提高免疫力等几个方向对其药理作用进行综述,为相关疾病的药物防治研究与临床应用提供理论依据。《实用医院临床杂志》,肖妤等,“红景天苷药理研究进展”,2006,3(3):98-99,红景天药理作用具有抗疲劳,抗衰老,免疫调节,清除自由基,抗肝纤维化,抗肾损害,改善保护心脑功能,抗肿瘤,抗辐射等多方面药理作用,应用前景广阔。
在雪莲中药材的开发中,其质量控制尤为关键,特别是如何控制雪莲活性成分提取中杂质的含量是一个难题。芦丁、绿原酸、紫丁香苷和去氢木香内酯等成分是非雪莲花的特有期望成分,如何将它们有效地与药理活性成分分离对提取工艺提高了非常高的要求和挑战。
CN101041653B公开了一种提取与富集雪莲总黄酮的方法,该方法先对雪莲进行超临界CO2萃取,得到其中的易挥发以及热不稳成分,然后再用常规溶剂提取。经过超临界萃取后的雪莲黄酮成分更容易从植物体内释放出来。
CN102106890B公开了雪莲总倍半萜内酯的应用及制备方法,和雪莲总倍半萜内酯在制备抗缺氧药物中的应用,其可通过溶剂法、二氧化碳超临界萃取、石油醚回流提取总倍半萜内酯,实验发现并证明了雪莲总倍半萜内酯用于抗缺氧药物的制备中制备所得的药物具有明显的抗缺氧作用。
CN104397683A水母雪莲花精华物质的提取方法,包括如下步骤:(1)水母雪莲花的预处理;(2)将预处理得到的水母雪莲花和去离子水及豆腐混合,同煮2-10小时,漂去豆腐,过滤,得滤液i,将过滤后所得的残渣阴干;(3)将阴干后的残渣与米醋以拌匀,闷润6-10分钟,待米醋被吸收尽后,至炒制容器内,用文火炒干,取出晾凉;(4)将冷却的残渣碾碎,过20-40目筛,放置超临界萃取釜中萃取,萃取的温度为50-70℃,萃取的压力为20-40Mpa,以乙醇为夹带剂,用CO2循坏萃取2-5小时,萃取结束后在分离釜中出料口出料,得滤液ⅱ;(5)将滤液ⅰ和滤液ⅱ混合,用孔径为0.22μm的微孔滤膜过滤,得滤液ⅲ;(6)将滤液ⅲ在60℃以下旋转蒸发减压浓缩,浓缩到固形物含量为60-80%为止;(7)将浓缩液进行干燥,即得水母雪莲花提取物。
CN106511213A公开了一种具备植物提取液的化妆品,由下列重量百分比的成分混合组成:芦荟20-50%,春黄菊5-20%,雪莲花5-15%,茉莉花10-30%,茶叶10-20%;将上述成分按照重量百分比混合后,干燥粉碎,药粉进行二氧化碳超临界萃取,以无水乙醇为溶剂萃取,减压浓缩回收乙醇,得到醇提浓缩液;将二氧化碳超临界萃取后的药渣以浓度10%-70%的乙醇溶液为提取溶剂,经过加热提取和浓缩后,浓缩液过大孔树脂柱进行富集,用浓度为30%-90%酒精多次洗脱,洗脱液经过减压浓缩后,用1.3丁二醇溶解,过滤,最后与二氧化碳超临界萃取值得溶液混合,即得植物提取液。
CN102106889A公开了一种雪莲和雪莲提取物在制备抗缺氧药物中的应用及其雪莲提取物的制备方法,所述雪莲选自大苞雪莲、水母雪莲或苞叶雪莲中的一种,所述雪莲提取物为大苞雪莲提取物、水母雪莲提取物或苞叶雪莲提取物中的一种;所述雪莲提取物为雪莲的水提物或雪莲的乙醇提取物及其经***溶剂法萃取和大孔吸附树脂法提取所得的抗缺氧有效部位。
CN101209274A一种雪莲提取物,其特征在于按重量百分比含有0.5%至1.2%的紫丁香苷、7%至17%的绿原酸和8%至25%的芦丁。
“雪莲水溶性多糖提取工艺的研究”,刘春兰等,食品与生物技术学报,2007,26(3):37-40,比较了料液比、温度、提取时间等因素对新疆雪莲组织培养物中雪莲多糖提取率的影响,并给出雪莲水溶性多糖的最佳提取工艺。
综上可知,在现有技术中尚存在多种缺陷,特别是现有技术中雪莲的提取方法仍沿用的是传统提取方法,虽然已有采用超临界方法提取雪莲药理活性物质的报导,但是仍是沿用的常规的超临界萃取设备和工艺,提取物中药理活性成分含量较低,并且不期望的杂质含量偏高。此外,尚未见有将雪莲提取物、淫羊藿提取物和红景天提取物复配制备复方制剂的报导,或者通常都是将三者含有复杂成分的提取物即粗提取物混合,不仅效果差,而且协同作用也较差。本领域需要一种高效的获得高药理活性的雪莲提取物的提取设备和方法。
发明内容
为同时解决现有技术中存在的上述技术问题,本发明提供了一种雪莲超临界提取装置,使用该装置时可以有效提取获得药理活性成分,药理活性成分含量高且不期望的杂质含量低。上述雪莲提取物以及包含其的复方制剂具有高的抗炎功效、提高免疫力、防辐射、抗疲劳功效等,以及提高的综合药理活性。
更具体地,本发明提供了一种提高免疫力、抗炎、防辐射并且补肾、益精血的组合物,该中药组合物特别为包含雪莲提取物、淫羊藿提取物和红景天提取物的中药组合物,即,提供了一种提高免疫力、抗炎、防辐射并且补肾、益精血的雪莲淫羊藿红景天中药组合物,因此本发明的主题(即名称)亦可为一种提高免疫力、抗疲劳、防辐射的雪莲淫羊藿红景天中药组合物及其制备方法。为此,本发明提供了以下技术方案。
在本发明的一方面,提供了一种雪莲超临界提取装置,该装置包括:入口部分,提取部分,和出口部分。
就本发明而言,所述入口部分可包括:二氧化碳供给储罐(1),连接到压力计量器(5)的阀门(2,4,7),压缩机(3),二氧化碳储存容器(6),高压泵(8),盘管(9)和朝向提取池(11)的CO2入口阀(10)。
就本发明而言,所述提取部分可包括:提取池(11),与提取池连接的压力计(12),由发光灯泡(13)、用于使温度均匀化的风扇(14)和精密温度计组成的加热***,所述精密温度计与温度控制器(15)连接用于测量温度。
就本发明而言,所述出口部分可包括:提取池的出口阀(16),提取物收集器(17),热浴(18)和流量计(19)。
优选地,所述提取池由不锈钢合金制成。
更优选地,所述不锈钢合金的组成以重量百分比计为:0.3%≤Mn≤0.8%;3.0%≤Cr≤6.0%;4.0%≤Ni≤10.0%;1.0%≤Al≤2%;1.0%≤Ti≤1.5%,0.001%≤P≤0.02%,0.1%≤W≤0.2%;0.001%≤S≤0.005%;0.0001%≤N≤0.0060%;0.0001%≤C≤0.025%;0.0001%≤Si≤0.25;0.0001%≤O≤0.0050%,余量为Fe。
本发明人研究发现,在该合金中,Ti的加入可显著提高合金的强度,其使合金经高温变形再进行时效细化晶粒后可提高其断裂韧性,而Ni的加入能够提高合金的韧性。Al和Ni以及Al和W形成的金属间化合物,在合金中能够起到弥散强化作用和抗氧化作用,从而提高合金的强度和冲击韧性,例如比不加Ni、W的合金相比,强度提高30-50%。
本发明的提取池可以由所述合金通过如下方法制得:
(1)由原料铸锭先制成半成产品;
(2)然后在850-920℃下使所述半成产品进行等温热处理,等温时间为3-6小时,然后快速冷却至低于或等于-80℃的温度来进行深度冷却处理;
(3)在400-480℃下进行时效退火处理,时效处理时间为10-20小时。
本发明通过上述方法获得的提取池在极高压力下仍具有特别好的抗断裂性和韧性,并且在高压下不易于产生由CO2酸性气体引起的表面侵蚀,因此使得能够在较高压力和较低温度下进行超临界提取。
在本发明的另一方面,提供了使用上述提取装置提取雪莲有效成分的方法,该方法包括:(1)在入口部分使装置达到提取所需的压力条件;(2)在提取部分于选定的温度和压力条件下进行提取;和(3)在出口部分取出提取物。
在本发明的另一方面,提供了使用上述装置或者使用上述方法获得的雪莲提取物。
优选地,所述雪莲提取物中挥发油的含量低于1.2wt%。更优选地,所述雪莲提取物中芦丁、绿原酸、紫丁香苷和去氢木香内酯的总含量低于2.5wt%。最优选地,所述雪莲提取物中具有下式(a)所示母核的内酯化合物总含量大于65.0wt%:
本发明人研究发现,当雪莲提取物中的上述组分同时满足所述含量要求时,能够特别有效地确保雪莲提取物的药理活性,特别是抗炎和提高免疫力以及防辐射效果。研究表明,所述内酯化合物是雪莲提取物中具有较好上述功效的一类化合物。
在本发明中,优选地,提取池(11)的直径为6.1厘米、高度为18厘米、有效容量为0.50L;所述发光灯泡功率为200-300W;所述温度计为ASQ F200精密温度计,所述温度控制器为基于单片机的温度控制器。在本发明中,由于前述合金制成的提取池可以承受较高的压力,因此提取可以在较低温度下进行,通过使用灯泡加热即可得到所需温度,从而能够克服现有常规加热手段例如油浴或水浴加热存在的加热不均匀和加热控制精度偏低(如飞温现象)的问题,从而能够有效防止超临界萃取条件的波动,有效减少提取副产物的产生。
本发明人经过大量实验发现,雪莲提取的最佳工艺条件为:将干燥粉碎(例如粒径400±25μm)的雪莲花(例如25克)放入超声波萃取池中,在40℃和压力10.34MPa下萃取出提取物,每批提取进行120-180分钟,二氧化碳排流速为125.25±15.00m L/min,将用于收集提取物的池浸在冰水浴(8±0.5℃)中。
在本发明的另一方面,提供了一种包含上述提取物的组合物。所述组合物还任选包含淫羊藿提取物和红景天提取物。雪莲提取物、淫羊藿提取物和红景天提取物的重量比为(10-5):(1-3):(1-3)。
在本发明的另一方面,提供了上述组合物的用途,其用于制备可增强人体免疫功能的药品、保健品或饮品。
优选地,淫羊藿组合物中的糖苷类物质的纯度大于99.0wt%。更优选地,所述糖苷类物质的纯度大于99.2wt%。最优选地,所述糖苷类物质的纯度大于99.7wt%。
所述糖苷类物质优选为或者包括下式(I)和(II)所示化合物的混合物:
式(I)和式(II)化合物的重量比优选为1:5-5:1。
研究表明,所述两种化合物为糖苷类化合物,与一般黄酮类化合物例如淫羊藿苷A和B相比,其具有特别广泛和相对较高的生物活性,如抗骨质疏松症、抗炎和抗癌以及补肾、益精血作用。
“淫羊藿总黄酮对肾阳虚大鼠下丘脑-垂体-肾上腺轴钙调蛋白基因表达的影响”,蒋淑君等,中国应用生理学杂志,2004;20(4),从分子水平探讨肾阳虚证病理改变及淫羊藿总黄酮的作用及其机理,为开发药物的临床新用途提供理论依据。在包括该文献在内的很多文献,都报道了包括上述淫羊藿黄酮化合物(亦为糖苷类化合物)的补肾、益精血功效,但是基本都是作为总黄酮(即总体混合物)进行研究。
吉林农业大学硕士学位论文,“朝鲜淫羊藿化学成分研究”,王斌,20070601,和北京化工大学硕士学位论文,“四川淫羊藿化学成分研究和抑菌活性研究”,宋丽娜,20090601,都报导了包括上述式(I)和式(II)化合物在内的众多黄酮化合物(即上述糖苷化合物)具有补肾、益精血功效,具体而言,所述淫羊藿苷能明显促进大鼠***TS基础分泌,这为淫羊藿用于TS水平低下的***的治疗提供了实验依据。淫羊藿对糖皮质激素所致“阳虚证”有明显的改善,各项指标恢复较快。但是,上述文献未具体指出上述两种具体的化合物具有更优的功效。
“淫羊藿属植物化学成分及药理研究进展”,谢娟平,海峡药学,2006年第18卷第5期,从药理角度报导了包括上述式(I)和式(II)化合物在内的众多黄酮化合物有益精气、坚筋骨、补腰肾、强心力的功效。同样,也未公开上述两种化合物具有更佳的补肾、益精血功效。
本发明人经过筛选,发现上述式(I)和式(II)化合物相对于提取物中的其它化合物而言,具有更佳的功效,将其作为目标化合物,采取特定的工艺进行提取,来制备前述组合物。
所述式(I)和式(II)化合物优选通过下面方法提取得到:
(1)在室温下用MeOH(优选3×6L,每次浸泡时间为约20h)提取淫羊藿草药粉(优选1.18kg),将合并的提取物减压蒸发得到棕色残余物(162.5g)即甲醇提取液;
(2)利用反相硅胶柱(例如RP-18硅胶柱),首先使用MeOH-H2O混合溶液对所述残余物进行梯度洗脱,来进行柱色谱分离,优选的洗脱步骤为0:1(1L),1:9(1L),2:8(1L),3:7(1L),4:6(1L),5:5(2L),6:4(2L),7:3(1L),1:0(2L),然后使用丙酮(2L),依次得到10个洗脱组分,记为洗脱组份A-J;
(3)将洗脱组分B(例如6.5g)在硅胶柱(优选60cm×5cm)上进行色谱分离,使用用CHCl3-MeOH-H2O混合溶液进行梯度洗脱(洗脱梯度优选为20:1:0(1L),10:1:0(2L),30:8:1(2L)和15:6:1(2.5L)),得到4个洗脱组份,记为洗脱组分B1-B4;
(4)将洗脱组分B2(例如300.2mg)用MeOH-H2O(1:1)在Sephadex LH-20(优选100cm×3cm)上分离,得到7个次洗脱组分,记为洗脱组分B2.1-B2.7;
(5)将洗脱组分B2.5和B2.6的混合物(例如总重量210.5mg)通过硅胶CC柱(优选80cm×2.5cm),洗脱液为EtOAc-CHCl3-MeOH-H2O(12:6:3:1),将洗脱液减压浓缩至干,得到淡黄色粉末(例如11.2mg),即为式(I)和式(II)所示化合物的混合物。
所述方法能够有效排除蛋白和其它杂质的干扰,另外一些低效价多糖也能得到有效的分离。本申请人通过上述5级柱层析分离,首次获得了极高纯度的式(I)和式(II)所示化合物,所述5级分离***之间具有密切的层级和配合关系,例如顺序的选择就非常重要。
就本发明而言,所述红景天提取物优选为红景天苷。红景天苷可以采取本领域的常规方法进行提取。更优选,所述红景天提取物为改性的红景天提取物,最优选地,所述红景天提取物为通过对红景天苷改性获得的下式(II)或(III)所示的化合物:
与一般天然苷类物质相比,式(II)或(III)所示的化合物对正常小鼠的免疫调节作用更明显,由于吸电子基团Cl和F的引入,增强了其清除自由基的能力,可显著增加小鼠特异的抗体分泌细胞数,在50mg/(kg·d)剂量即可增强迟发型超敏反应强度,另外还表现出对异型小鼠的混合淋巴细胞反应及巨噬细胞的吞噬功能,并且在一定程度上可以降低白细胞介素的活性,表明其二者是一种很有前途的免疫调节剂。
具有结构式(II)-(III)的物质可以通过如下方法合成:在反应容器中加入MCM-22分子筛作为选择性催化剂,然后加入氯仿溶剂,再将天然红景天苷与氯代或氟代的乙酰氯以1:1-1.3的摩尔比加入到反应溶剂中(天然红景天苷与氯代或氟代的乙酰氯优选均以氯仿溶液形式加入),反应温度为20-30℃,反应时间为10-30min,分离和干燥即可获得所述化合物。该方法具有收率高、杂质少的优点。尤其是由于MCM-22分子筛的择形作用,可显著降低副反应的发生。
在本发明的另一方面,提供了所述组合物的用途,其用于制备可增强人体免疫功能的具有抗炎效果、抗疲劳和防辐射效果的药品、保健品或饮品。
优选地,所述药品为复方中药制剂。优选地,所述组合物用于制备口服胶囊、冲剂等。所述保健品包括口腔用品例如咀嚼片或含片。
附图说明
图1是根据本发明的提取装置的示意图。
具体实施方式
实施例1
用于进行提取的雪莲为天山雪莲(购自乌鲁木齐北园春农副产品经营有限公司)。采用的提取装置包括:入口部分,提取部分,和出口部分,所述入口部分可包括:二氧化碳供给储罐(1),连接到压力计量器(5)的阀门(2,4,7),压缩机(3),二氧化碳储存容器(6),高压泵(8),盘管(9)和朝向提取池(11)的CO2入口阀(10);所述提取部分可包括:提取池(11),与提取池连接的压力计(12),由发光灯泡(13)、用于使温度均匀化的风扇(14)和精密温度计组成的加热***,所述精密温度计与温度控制器(15)连接用于测量温度,所述出口部分可包括:提取池的出口阀(16),提取物收集器(17),热浴(18)和流量计(19);所述提取池由上文所述不锈钢合金制成;工艺条件为:将25克干燥粉碎(400±25μm)的雪莲花放入超声波萃取池中,在40℃和10.34MPa压力下萃取出提取物,每批提取进行180分钟,二氧化碳排流速为125.25±17.22mL/min,将用于收集提取物的池浸在冰水浴(8±0.5℃)中。经检测,雪莲提取物中挥发油的含量为1.0wt%,芦丁、绿原酸、紫丁香苷和去氢木香内酯的总含量为1.1wt%。最优选地,所述雪莲提取物中具有式(a)所示母核的内酯化合物总含量为81.1wt%。
对比例1
按照CN102106890B中实施例记载的方法进行超临界提取。经检测,雪莲提取物中挥发油的含量为2.3wt%,芦丁、绿原酸、紫丁香苷和去氢木香内酯的总含量为4.3wt%。最优选地,所述雪莲提取物中具有式(a)所示母核的内酯化合物总含量为77.3wt%。
由上述实施例和对比例可以看出,本发明的提取装置能够在内酯化合物总含量稍有提高的情况下,可以显著降低不期望的组分的含量。
实施例2
测试实施例1的雪莲提取物对佐剂性关节炎继发性损害的抑制作用。
取Wistar大鼠60只,雌雄各半,体重150~200g,雌雄各半,随机分为3组,即正常对照组,常规提取物组(0.05g·kg-1,对照组,其中雪莲提取物按照CN101041653B中记载的方法获得),实施例1提取物组(0.05g·kg-1);先用足趾容积测量仪测量每组大鼠左后足跖容积作为基础值。除正常对照组外,其余各组均在右后足跖皮下注射Freund’s完全佐剂0.1mL,致炎后第12d各给药组开始灌胃给药,正常对照组和模型组灌胃等量的蒸馏水,连续8d。给药后各组均隔天以足趾容积仪测量左后足跖(非注射足趾)容积至第20d。计算各组动物的足趾肿胀度。
与模型组和对照组相比,实施例1的雪莲提取物对佐剂性关节炎继发性损害抑制作用有显著性差异,表明本发明的复方雪莲胶囊能有效减轻佐剂性关节炎继发性损害,有明显的抗炎作用,即使低剂量的情况下,随着给药时间延长也有明显抑制作用,见下表1:
表1:雪莲提取物对佐剂性关节炎继发性损害抑制作用
由上面实施例清楚地可以看出,本发明的雪莲提取物相对于常规超临界提取法得到的提取物具有良好的抑制作用,特别是具有明显更好的抗炎效果。
实施例3
包含雪莲提取物、淫羊藿提取物和红景天提取物的胶囊的制备,其中取实施例1所述的雪莲提取物、前文所述的淫羊藿提取物和红景天提取物,雪莲提取物、淫羊藿提取物和红景天提取物的重量比为8:1:1,每个胶囊含有0.1g内容物。将该胶囊按照实施例2的方法进行对佐剂性关节炎继发性损害的抑制作用测试,在相同剂量和实验条件下,致炎后第3天的足趾肿胀度(ml)为0.86±0.11。
由实施例2和3同样可以看出,淫羊藿提取物和红景天提取物的加入,增强了雪莲提取物的抗炎效果,进而具有较好的提高免疫力和抗疲劳效果。推测其原因,除了淫羊藿提取物和红景天提取物本身具有一定的抗炎效果外,三者之间可能还产生了一定相互协同促进作用。
本书面描述使用实例来公开本发明,包括最佳模式,且还使本领域技术人员能够制造和使用本发明。本发明的可授予专利的范围由权利要求书限定,且可以包括本领域技术人员想到的其它实例。如果这种其它实例具有不异于权利要求书的字面语言的结构元素,或者如果这种其它实例包括与权利要求书的字面语言无实质性差异的等效结构元素,则这种其它实例旨在处于权利要求书的范围之内。在不会造成不一致的程度下,通过参考将本文中参考的所有引用之处并入本文中。
Claims (1)
1.一种可提高免疫力、抗炎、防辐射雪莲淫羊藿红景天组合物的制备方法,该组合物包含淫羊藿提取物和红景天提取物,雪莲提取物、淫羊藿提取物和红景天提取物的重量比为(10-5):(1-3):(1-3);
所述雪莲提取物通过使用下述提取装置得到,该提取装置包括:入口部分,提取部分,和出口部分,所述入口部分包括:二氧化碳供给储罐,连接到压力计量器的阀门,压缩机,二氧化碳储存容器,高压泵,盘管和朝向提取池的CO2入口阀;雪莲提取的工艺条件为:将干燥粉碎的雪莲花放入超声波萃取池中,在40℃和压力10.34MPa下萃取出提取物,每批提取进行120-180分钟,二氧化碳排流速为125.25±15.00m L/min,将用于收集提取物的池浸在冰水浴即8±0.5℃中;所述雪莲提取物中挥发油的含量低于1.2wt%,所述雪莲提取物中芦丁、绿原酸、紫丁香苷和去氢木香内酯的总含量低于2.5wt%,所述雪莲提取物中具有下式(a)所示母核的内酯化合物总含量大于65.0wt%:
所述淫羊藿提取物中的糖苷类物质的纯度大于99.0wt%;
所述糖苷类物质为下式(I)和(II)所示化合物的混合物:
式(I)和式(II)化合物的重量比为1:5-5:1;
所述红景天提取物为通过对红景天苷改性获得的下式(III)所示的化合物:
所述式(III)所示的化合物通过如下方法合成:在反应容器中加入MCM-22分子筛作为选择性催化剂,然后加入氯仿溶剂,再将天然红景天苷与氯代的乙酰氯以1:1-1.3的摩尔比加入到反应溶剂中,反应温度为20-30℃,反应时间为10-30min,分离和干燥即获得所述式(III)所示的化合物;
其中所述式(I)和式(II)化合物通过下面方法提取得到:
(1)在室温下用MeOH提取淫羊藿草药粉,将合并的提取物减压蒸发得到棕色残余物即甲醇提取液;
(2)利用反相硅胶柱,首先使用MeOH-H2O混合溶液对所述残余物进行梯度洗脱,来进行柱色谱分离,洗脱步骤为0:1共1L,1:9共1L,2:8共1L,3:7共1L,4:6共1L,5:5共2L,6:4共2L,7:3共1L,1:0共2L,然后使用丙酮2L,依次得到10个洗脱组分,记为洗脱组份A-J;
(3)将洗脱组分B在硅胶柱上进行色谱分离,使用CHCl3-MeOH-H2O混合溶液进行梯度洗脱,洗脱梯度为20:1:0共1L,10:1:0共2L,30:8:1共2L和15:6:1共2.5L,得到4个洗脱组份,记为洗脱组分B1-B4;
(4)将洗脱组分B2用1:1的MeOH-H2O在Sephadex LH-20上分离,得到7个次洗脱组分,记为洗脱组分B2.1-B2.7;
(5)将洗脱组分B2.5和B2.6的混合物通过硅胶CC柱,洗脱液为12:6:3:1的EtOAc-CHCl3-MeOH-H2O,将洗脱液减压浓缩至干,得到淡黄色粉末,即为式(I)和式(II)所示化合物的混合物。
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