CN105687497A - 含莲子心黄酮类化合物的组合物及其在降血脂方面的应用 - Google Patents
含莲子心黄酮类化合物的组合物及其在降血脂方面的应用 Download PDFInfo
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Abstract
本发明公开了一种含莲子心黄酮类化合物的组合物及其在降血脂方面的应用,属于医药保健品技术领域。其采用微波-超声波协同辅助萃取法获得莲子心总黄酮,然后经聚酰胺树脂柱层析纯化后得到莲子心黄酮类化合物,其芦丁含量达77.8mg/g,再将所得莲子心黄酮类化合物与制得的灵芝多糖、桑黄多糖及茶多酚复配制成所述组合物。经动物药效学验证,本发明组合物可明显降低高脂动物的甘油三酯和总胆固醇水平,将其制成胶囊剂、片剂、丸剂、散剂、口服液、颗粒剂等常见剂型,与临床治疗药物配合使用,能起到显著辅助降血脂的功能。
Description
技术领域
本发明属于医药保健品技术领域,具体涉及一种含莲子心黄酮类化合物的组合物及其在降血脂方面的应用。
背景技术
高脂血症主要是指由脂肪代谢或转运异常导致血浆中一种或多种脂质浓度升高的症状。由于高脂血症的类型不同,降血脂药物根据其药理作用分为降低甘油三酯、降低胆固醇两大类。
随着生活水平的逐渐提高,高血脂症的发生率呈逐年上升趋势,高血脂症是导致冠心病、动脉硬化、心肌梗死发生的重要病因之一,还会加重糖尿病、高血压、胰腺炎、肝硬化等疾病的进展。目前用于高血脂症临床治疗的药物以西药为主,虽然其作用效果明显,但有副作用,易引起机体的不良反应。我国降血脂药物的市场及研发处于起步状态,发展降血脂药物、中成药制剂以及保健品,是我国对于降血脂研究的主要方向。
莲子心有广泛的药理作用,在传统中医药中占有重要地位,为《中国药典》所收录。黄酮为莲子心的主要活性成分,是莲子心广泛药理作用的主要物质基础。本发明对莲子心黄酮类化合物进行提取与富集纯化,进一步开发出具有降脂功效的药物和保健食品。
发明内容
本发明的目的在于提供一种含莲子心黄酮类化合物的组合物及其在降血脂方面的应用,该组合物可进一步制备成具有降血脂功效的药物或保健品,与临床治疗药物配合使用,能起到显著辅助降血脂的功能,并可降低药物的毒副作用。
为实现上述目的,本发明采用如下技术方案:
一种含莲子心黄酮类化合物的组合物,其制备方法包括以下步骤:
1)称取新鲜莲子心、灵芝及桑黄,洗去杂质,40-50℃烘干至恒重后,分别超微粉碎至细度为0.1-1.5μm,得莲子心超微粉、灵芝超微粉及桑黄超微粉;
2)向步骤1)制备的莲子心超微粉中加入质量浓度为70%-80%的乙醇溶液,微波-超声波协同辅助提取10-20min,过滤;将滤渣按上述步骤重复提取2-3次,合并滤液,于3000-4000r/min条件下离心10-20min,取上清液,在50-65℃条件下浓缩至浸膏状,冷冻干燥后粉碎,得莲子心总黄酮;
3)将步骤2)所得莲子心总黄酮用质量浓度为70%的乙醇溶液溶解后,采用60-80目聚酰胺树脂进行柱层析,以质量浓度为40%-60%的乙醇溶液为洗脱剂,将所得洗脱液经浓缩、冷冻干燥,得到纯化的莲子心黄酮类化合物;
4)将步骤3)所得莲子心黄酮类化合物粉碎并过80-100目筛,备用;
5)按重量体积之比为1:20-30在步骤1)制备的灵芝超微粉中加入水,于80-95℃下提取30-60min,过滤,取上清液;将滤渣依照上述步骤重复提取2-3次,合并滤液,在滤液中加入其体积3-4倍、质量浓度为95%的乙醇溶液,沉淀6-10h后,4000-6000r/min离心收集沉淀,经冷冻干燥后粉碎,过50-80目筛,得灵芝多糖;
6)按重量体积之比为1:10-15在步骤1)制备的桑黄超微粉中加入水,于60-85℃下提取90-120min,过滤,取上清液;将滤渣依照上述步骤重复提取2-3次,合并滤液,在滤液中加入其体积3-4倍、质量浓度为95%的乙醇溶液,沉淀8-12h后,4000-6000r/min离心收集沉淀,经冷冻干燥后粉碎,过50-80目筛,得桑黄多糖;
7)将步骤4)所得粉碎后的莲子心黄酮类化合物、步骤5)所得灵芝多糖、步骤6)所得桑黄多糖与天然活性物质茶多酚混合搅拌20-30min制得。
步骤2)中所用莲子心超微粉的重量与乙醇溶液的体积之比为1:20-40;
所述微波-超声波协同辅助提取中微波功率为300-400W,超声功率为200-400W,超声程序为运行1s后停止0~2s,如此循环。
步骤3)中粉碎后的物料重量与加入水的体积之比为1:0.5-1.5。
步骤7)中所用原料按重量份计为:莲子心黄酮类化合物5-10份、灵芝多糖1-2份、桑黄多糖1-2份、茶多酚2-4份。
所述含莲子心黄酮类化合物的组合物可与常规辅料配合,用于制备具有降血脂功效的药物或保健品;所述药物或保健品为药剂学上接受的剂型,包括胶囊剂、片剂、丸剂、散剂、口服液。
本发明的显著优点在于:本发明所得莲子心黄酮类化合物中芦丁含量达77.8mg/g,将其与灵芝多糖、桑黄多糖、茶多酚等天然活性物质配合作为组合物,与临床治疗药物配合使用,能起到显著辅助降血脂的功能,并可降低药物的毒副作用。本发明所用原料来源符合药食同源标准,其是一种极具开发前景的功能性保健食品。
附图说明
图1为不同处理对小鼠甘油三酯水平的影响。
图2为不同处理对小鼠总胆固醇水平的影响。
具体实施方式
为了使本发明所述的内容更加便于理解,下面结合具体实施方式对本发明所述的技术方案做进一步的说明,但是本发明不仅限于此。
实施例1
一种含莲子心黄酮类化合物的组合物,其制备方法包括以下步骤:
1)称取新鲜莲子心、灵芝及桑黄,洗去杂质,40℃烘干至恒重后,分别超微粉碎至细度为0.1μm,得莲子心超微粉、灵芝超微粉及桑黄超微粉;
2)向步骤1)制备的莲子心超微粉中按重量体积比1:20(g/mL)加入质量浓度为80%的乙醇溶液,微波-超声波协同辅助提取10min,微波功率为300W,超声功率为200W,超声程序为运行1s后停止0~2s,如此循环,然后过滤;将滤渣按上述步骤重复提取2次,合并滤液,于3000r/min条件下离心20min,取上清液,在50℃条件下浓缩至浸膏状,冷冻干燥后粉碎,得莲子心总黄酮;
3)将步骤2)所得莲子心总黄酮按重量体积比1:0.5(g/mL)加质量浓度为70%的乙醇溶液溶解,采用60目聚酰胺树脂进行柱层析,以质量浓度为40%的乙醇溶液为洗脱剂,将所得洗脱液经浓缩、冷冻干燥,得到纯化的莲子心黄酮类化合物;
4)将步骤3)所得莲子心黄酮类化合物粉碎并过80目筛,备用;
5)按重量体积之比为1:20(g/mL)在步骤1)制备的灵芝超微粉中加入水,于80℃下提取60min,过滤,取上清液;将滤渣依照上述步骤重复提取2次,合并滤液,在滤液中加入其体积3倍、质量浓度为95%的乙醇溶液,沉淀10h后,4000r/min离心收集沉淀,经冷冻干燥后粉碎,过50目筛,得灵芝多糖;
6)按重量体积之比为1:10(g/mL)在步骤1)制备的桑黄超微粉中加入水,于60℃下提取120min,过滤,取上清液;将滤渣依照上述步骤重复提取2次,合并滤液,在滤液中加入其体积3倍、质量浓度为95%的乙醇溶液,沉淀12h后,4000r/min离心收集沉淀,经冷冻干燥后粉碎,过50目筛,得桑黄多糖;
7)将步骤4)所得粉碎后的莲子心黄酮类化合物5重量份、步骤5)所得灵芝多糖1重量份、步骤6)所得桑黄多糖1重量份与天然活性物质茶多酚2重量份混合搅拌20min制得。
实施例2
一种含莲子心黄酮类化合物的组合物,其制备方法包括以下步骤:
1)称取新鲜莲子心、灵芝及桑黄,洗去杂质,45℃烘干至恒重后,分别超微粉碎至细度为1.0μm,得莲子心超微粉、灵芝超微粉及桑黄超微粉;
2)向步骤1)制备的莲子心超微粉中按重量体积比1:30(g/mL)加入质量浓度为75%的乙醇溶液,微波-超声波协同辅助提取15min,微波功率为350W,超声功率为300W,超声程序为运行1s后停止0~2s,如此循环,然后过滤;将滤渣按上述步骤重复提取2次,合并滤液,于3500r/min条件下离心15min,取上清液,在60℃条件下浓缩至浸膏状,冷冻干燥后粉碎,得莲子心总黄酮;
3)将步骤2)所得莲子心总黄酮按重量体积比1:1(g/mL)加质量浓度为70%的乙醇溶液溶解,采用80目聚酰胺树脂进行柱层析,以质量浓度为50%的乙醇溶液为洗脱剂,将所得洗脱液经浓缩、冷冻干燥,得到纯化的莲子心黄酮类化合物;
4)将步骤3)所得莲子心黄酮类化合物粉碎并过80目筛,备用;
5)按重量体积之比为1:25(g/mL)在步骤1)制备的灵芝超微粉中加入水,于90℃下提取40min,过滤,取上清液;将滤渣依照上述步骤重复提取3次,合并滤液,在滤液中加入其体积4倍、质量浓度为95%的乙醇溶液,沉淀8h后,5000r/min离心收集沉淀,经冷冻干燥后粉碎,过60目筛,得灵芝多糖;
6)按重量体积之比为1:12(g/mL)在步骤1)制备的桑黄超微粉中加入水,于70℃下提取100min,过滤,取上清液;将滤渣依照上述步骤重复提取3次,合并滤液,在滤液中加入其体积3倍、质量浓度为95%的乙醇溶液,沉淀10h后,5000r/min离心收集沉淀,经冷冻干燥后粉碎,过60目筛,得桑黄多糖;
7)将步骤4)所得粉碎后的莲子心黄酮类化合物8重量份、步骤5)所得灵芝多糖1重量份、步骤6)所得桑黄多糖1重量份与天然活性物质茶多酚3重量份混合搅拌25min制得。
实施例3
一种含莲子心黄酮类化合物的组合物,其制备方法包括以下步骤:
1)称取新鲜莲子心、灵芝及桑黄,洗去杂质,40-50℃烘干至恒重后,分别超微粉碎至细度为1.5μm,得莲子心超微粉、灵芝超微粉及桑黄超微粉;
2)向步骤1)制备的莲子心超微粉中按重量体积比1:40(g/mL)加入质量浓度为70%的乙醇溶液,微波-超声波协同辅助提取20min,微波功率为400W,超声功率为400W,超声程序为运行1s后停止0~2s,如此循环,然后过滤;将滤渣按上述步骤重复提取3次,合并滤液,于4000r/min条件下离心10min,取上清液,在65℃条件下浓缩至浸膏状,冷冻干燥后粉碎,得莲子心总黄酮;
3)将步骤2)所得莲子心总黄酮按重量体积比1:1.5(g/mL)加质量浓度为70%的乙醇溶液溶解,采用80目聚酰胺树脂进行柱层析,以质量浓度为60%的乙醇溶液为洗脱剂,将所得洗脱液经浓缩、冷冻干燥,得到纯化的莲子心黄酮类化合物;
4)将步骤3)所得莲子心黄酮类化合物粉碎并过100目筛,备用;
5)按重量体积之比为1:30(g/mL)在步骤1)制备的灵芝超微粉中加入水,于95℃下提取30min,过滤,取上清液;将滤渣依照上述步骤重复提取3次,合并滤液,在滤液中加入其体积4倍、质量浓度为95%的乙醇溶液,沉淀6h后,6000r/min离心收集沉淀,经冷冻干燥后粉碎,过80目筛,得灵芝多糖;
6)按重量体积之比为1:15(g/mL)在步骤1)制备的桑黄超微粉中加入水,于85℃下提取90min,过滤,取上清液;将滤渣依照上述步骤重复提取3次,合并滤液,在滤液中加入其体积4倍、质量浓度为95%的乙醇溶液,沉淀8h后,6000r/min离心收集沉淀,经冷冻干燥后粉碎,过80目筛,得桑黄多糖;
7)将步骤4)所得粉碎后的莲子心黄酮类化合物10重量份、步骤5)所得灵芝多糖2重量份、步骤6)所得桑黄多糖2重量份与天然活性物质茶多酚4重量份混合搅拌30min制得。
本发明含莲子心黄酮类化合物的组合物对高血脂模型小鼠血清甘油三酯和总胆固醇的影响
采用健康成年雄性昆明种小鼠为试验动物,动物适应性饲养一周后无不良反应,开始试验。高脂高能饲料喂养一个月造模后,将其随机分为1个阳性药物对照组、1个高血脂模型组和1个组合物实验组,每组10只,组合物实验组按100mg/(kg·d)进行灌胃给药,阳性药物对照组按100mg/(kg·d)给予盐酸二甲双弧,高血脂模型组给予等剂量生理盐水。在连续给药第21天时,对各组试验小鼠禁食不禁水8h,眼内眦取血,置离心管中于4℃条件下、12000r/min离心15min,分离血清,精密吸取血清100μL,按试剂盒说明操作,分别按式(1)和式(2)计算小鼠血清甘油三酯(TG)和胆固醇(TC)含量。
TG(mmol/L)=式(1)
TC(mmol/L)=式(2)
图1、图2分别为不同处理对小鼠甘油三酯水平及总胆固醇水平的影响。从图中可以看出,实验组小鼠的血清甘油三酯TG水平和总胆固醇TC水平都与模型组小鼠有显著差异,证明本发明所得含莲子心黄酮类化合物的组合物降脂效果好,可作为具有降脂功能的食品、药品、保健品原料。
以上所述仅为本发明的较佳实施例,凡依本发明申请专利范围所做的均等变化与修饰,皆应属本发明的涵盖范围。
Claims (5)
1.一种含莲子心黄酮类化合物的组合物,其特征在于:其制备方法包括以下步骤:
1)称取新鲜莲子心、灵芝及桑黄,洗去杂质,40-50℃烘干至恒重后,分别超微粉碎至细度为0.1-1.5μm,得莲子心超微粉、灵芝超微粉及桑黄超微粉;
2)向步骤1)制备的莲子心超微粉中加入质量浓度为70%-80%的乙醇溶液,微波-超声波协同辅助提取10-20min,过滤;将滤渣按上述步骤重复提取2-3次,合并滤液,于3000-4000r/min条件下离心10-20min,取上清液,在50-65℃条件下浓缩至浸膏状,冷冻干燥后粉碎,得莲子心总黄酮;
3)将步骤2)所得莲子心总黄酮用质量浓度为70%的乙醇溶液溶解后,采用60-80目聚酰胺树脂进行柱层析,以质量浓度为40%-60%的乙醇溶液为洗脱剂,将所得洗脱液经浓缩、冷冻干燥,得到纯化的莲子心黄酮类化合物;
4)将步骤3)所得莲子心黄酮类化合物粉碎并过80-100目筛,备用;
5)按重量体积之比为1:20-30在步骤1)制备的灵芝超微粉中加入水,于80-95℃下提取30-60min,过滤,取上清液;将滤渣依照上述步骤重复提取2-3次,合并滤液,在滤液中加入其体积3-4倍、质量浓度为95%的乙醇溶液,沉淀6-10h后,4000-6000r/min离心收集沉淀,经冷冻干燥后粉碎,过50-80目筛,得灵芝多糖;
6)按重量体积之比为1:10-15在步骤1)制备的桑黄超微粉中加入水,于60-85℃下提取90-120min,过滤,取上清液;将滤渣依照上述步骤重复提取2-3次,合并滤液,在滤液中加入其体积3-4倍、质量浓度为95%的乙醇溶液,沉淀8-12h后,4000-6000r/min离心收集沉淀,经冷冻干燥后粉碎,过50-80目筛,得桑黄多糖;
7)将步骤4)所得粉碎后的莲子心黄酮类化合物、步骤5)所得灵芝多糖、步骤6)所得桑黄多糖与天然活性物质茶多酚混合搅拌20-30min制得。
2.根据权利要求1所述含莲子心黄酮类化合物的组合物,其特征在于:步骤2)中所用莲子心超微粉的重量与乙醇溶液的体积之比为1:20-40;
所述微波-超声波协同辅助提取中微波功率为300-400W,超声功率为200-400W,超声程序为运行1s后停止0~2s,如此循环。
3.根据权利要求1所述含莲子心黄酮类化合物的组合物,其特征在于:步骤3)中粉碎后的物料重量与加入水的体积之比为1:0.5-1.5。
4.根据权利要求1所述含莲子心黄酮类化合物的组合物,其特征在于:步骤7)中所用原料按重量份计为:莲子心黄酮类化合物5-10份、灵芝多糖1-2份、桑黄多糖1-2份、茶多酚2-4份。
5.一种如权利要求1所述含莲子心黄酮类化合物的组合物的应用,其特征在于:将所得组合物与常规辅料配合,用于制备具有降血脂功效的药物或保健品。
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