CN104367659A - 一种稠李提取物的制备方法及在降血脂药品及保健品中的应用 - Google Patents
一种稠李提取物的制备方法及在降血脂药品及保健品中的应用 Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- A—HUMAN NECESSITIES
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract
本发明属于医药技术领域,主要公开了稠李提取物的制备方法及在药品和保健品中的新用途。经实验研究表明:稠李提取物对高脂饮食诱导的高血脂小鼠具有明显的降血脂作用。因此,可以使用该提取物制备降血脂药物、保健品、食品添加剂或复方制剂等。其药品或保健品可以是现有的任何剂型,如片剂,胶囊剂,粉针剂,注射剂,丸剂,软胶囊,颗粒剂和贴剂等。
Description
技术领域
本发明涉及一种稠李提取物的制备及在医药和保健品中的新用途;属于中药提取物的新生物活性的医药技术领域。
背景技术
稠李属植物在我国种类繁多,为多年生乔木,在我国广泛种植,因耐寒耐阴在东北地区成长密集。主要包括山桃稠李(Padus maackii),星毛稠李(Padus buergeriana),稠李(Padus racemosa),褐毛稠李(Padus brunnescens)以及紫叶稠李(Padus virginiana)等。因该属植物整体美观,在我国是一种重要的观赏物种。其中山桃稠李,稠李和紫叶稠李为蔷薇科李属植物,在东北分布较广。
研究表明:稠李属植物的叶子可以用于制作产品(稠李叶茶及其制备方法,专利,ZL. 200610010198.7),花因含有大量的苯甲醛可以做防腐用料(赵秋雁,稠李Prunus padus Linn.挥发油化学组成分析,《植物研究》,2003),此外,稠李含有的丰富黄酮类成分(何映霞,紫叶稠李叶片中黄酮的提取工艺优化《北方园艺》,2013年)和花色苷类成分(辛越,稠李属三种果实花色苷抗氧化活性研究,东北林业大学硕士论文,2013年)。上述文献报道了稠李的抗氧化和对HepG2的细胞毒活性(冷梅,紫叶稠李果实花色苷的抗氧化活性,食品科学,2013年;辛越,稠李属三种果实花色苷对HepG2细胞的增殖抑制作用比较,食品工业科技,2013年)。
到目前为止关于稠李的提取物制备方法未见文献报道,且未有关该提取物在降血脂方面的研究报道。本发明创新性的提出了稠李提取物的制备方法及在降血脂方面的新用途。
发明内容
本发明提供了一种稠李提取物的制备方法及在降血脂的药物或保健品及食品添加剂的应用。
一种稠李提取物的制备方法,通过如下步骤获得:干燥的稠李样品,粉碎过20目筛,经适当的溶剂和提取方法,得到提取液,减压浓缩至干;用水混悬至无沉淀,上大孔树脂吸附柱色谱,依次用水和70%以上的乙醇进行洗脱,收集醇洗脱部分,即得稠李提取物。
前述的稠李,选自山桃稠李(Padus maackii),稠李(Padus racemosa)或紫叶稠李(Padus virginiana)的任何一种;提取部位选自其果实,茎叶或花等任一部位,其优选果实。提取方法选自浸泡提取、回流提取、超声提取、逆流提取、超临界萃取等的任何一种;提取溶剂选自水,甲醇,乙醇任何一种及不同比例混合物;用于样品后续纯化处理的大孔吸附柱色谱,包括药学上能够接受的,可以用于分离的多种常规树脂,选自D101,NKA12或ADS等的任何一种。
前述的稠李提取物,主要包含黄酮,花色苷等抗氧化成分。
前述的稠李提取物,具有降血脂作用,可以用于药物、保健品及食品添加剂中的应用。
当本发明用于上述用途时,其口服或胃肠外给药,均是安全的,在口服情况下,其可以以任何常规的形式给药,如片剂,胶囊剂,粉针剂,注射剂,丸剂,软胶囊,颗粒剂和贴剂等。
本发明制备具有降血脂活性的桔梗提取物可以与药用载体或食品载体混合,这里使用的固体或液体的赋形剂在本领域是总所周知的,下面举几个例子,散剂是内服的粉末剂,它的赋形剂有乳糖,淀粉,糊精,碳酸钙,合成或天然硫酸铝,氧化镁,硬脂酸镁,碳酸氢钠,干燥酵母等;溶液剂的赋形剂有水,甘油,1,2-丙二醇,单糖浆,乙醇,乙二醇,聚乙二醇,山梨糖醇等;软膏剂的赋形剂可以使用脂油,含水羊毛脂、凡士林、甘油、蜂蜡、木蜡、液体石蜡等组合成的疏水剂或亲水剂。
本发明可以通过下面的实验例进一步说明。
实施例1. 稠李提取物的制备
制备方法1:取500克干燥的稠李果实(山桃稠李,Padus maackii),浸泡于70%的乙醇的水溶液中,用回流提取的方法提取3次,每次30min,提取液趁热过滤,滤液旋转蒸发除去溶剂。浓缩至干的浸膏在经水分散至无沉淀,经D101大孔吸附树脂柱色谱吸附,依次用水和70%以上乙醇解吸附,收集醇洗脱部分,减压浓缩至无醇味,经冷冻干燥得粉末状提取物。
制备方法2:取500克干燥的稠李叶子(山桃稠李,Padus maackii),浸泡于70%的乙醇的水溶液中,用回流提取的方法提取3次,每次30min,提取液趁热过滤,滤液旋转蒸发除去溶剂。浓缩至干的浸膏在经水分散至无沉淀,经D101大孔吸附树脂柱色谱吸附,依次用水和70%以上乙醇解吸附,收集醇洗脱部分,减压浓缩至无醇味,经冷冻干燥得粉末状提取物。
制备方法3:取500克干燥的稠李果实(紫叶稠李,Padus virginiana),浸泡于70%的乙醇的水溶液中,用超声波提取的方法提取3次,每次30min,提取液趁热过滤,滤液旋转蒸发除去溶剂。浓缩至干的浸膏在经水分散至无沉淀,经D101大孔吸附树脂柱色谱吸附,依次用水和70%以上乙醇解吸附,收集醇洗脱部分,减压浓缩至无醇味,经冷冻干燥得粉末状提取物。
制备方法4:取500克干燥的稠李果实(紫叶稠李,Padus virginiana),浸泡于70%的乙醇的水溶液中,用超声波提取的方法提取3次,每次30min,提取液趁热过滤,滤液旋转蒸发除去溶剂。浓缩至干的浸膏在经水分散至无沉淀,经D101大孔吸附树脂柱色谱吸附,依次用水和70%以上乙醇解吸附,收集醇洗脱部分,减压浓缩至无醇味,经冷冻干燥得粉末状提取物。
实施例2. 稠李提取物中总黄酮的测定
1.1 实验材料
前述的稠李提取物,来源为紫叶稠李(Padus virginiana)的果实。
1.2 仪器与试剂
电子天平;紫外可见分光光度计;真空干燥箱;水浴锅;RE-52AA型旋转蒸发仪;高速离心机;芦丁对照品(98.5%),实验室自制;无水乙醇,氢氧化钠;硝酸铝;亚硝酸钠等,北京化工厂。
1.3 方法
1.3.1 芦丁对照品溶液的制备
精密称取减压干燥至恒重的芦丁对照品10.00 mg,置于50ml 容量瓶中,加70% 的乙醇溶液,置于水浴上加热溶解,放冷;然后,用70%的乙醇溶液稀释至刻度线,摇匀即得0.20 mg/ml的标准溶液。分别取上述标准溶液0.5、1.0、1.5、2.0、2.5、3.0、4.0 ml于10ml量筒,各加30%乙醇,使成为5ml;先加5%亚硝酸钠溶液0.3ml,摇匀,放置6min;再加10%硝酸铝0.3ml,摇匀,放置6min;加4% 氢氧化钠4.0ml,再加30%乙醇溶液至10.0ml。则分别得到不同浓度的标准溶液。
1.3.2 标准曲线的制备
分别从上述制备的标准溶液中,以30%乙醇溶液5ml,5% 亚硝酸0.3ml,10%硝酸铝溶液0.3 ml,4%氢氧化钠4.0ml,然后再用30% 乙醇溶液定容至10.0ml 的溶液作空白液,用紫外分光光度计;以3 cm比色皿,510 nm波长测定吸光度,作以芦丁量(mg)与吸光度的标准曲线,用最小二乘法作线性回归,得芦丁量Y(mg)与吸光度X的方程。
1.3.3 总黄酮含量测定
移液管移取1ml样品于量筒,用70%乙醇稀释到所需溶液。使其颜色接近芦丁标准液颜色,尽量使其吸光度在0.2~0.6的范围内,以保证其吸光度检测稳定性。从已稀释好的样品中移取1ml于量筒。加30%乙醇至5.0ml,先加5%氢氧化钠0.3ml,摇匀,静置6min,再加10%硝酸铝0.3ml,摇匀静置6min,再加4%氢氧化钠4.0ml,用30%乙醇定容至10ml,摇匀,静置15min后测定。
2 实验结果
结果表明稠李提取中总黄酮含量为2.56%
实施例3. 稠李提取物对高脂血症大鼠的降血脂作用
1.1 实验材料
前述的稠李提取物,来源为紫叶稠李(Padus virginiana)的果实。
1.2 仪器及试剂
解剖器械、酶标仪,胆固醇、胆酸钠、血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C),高密度脂蛋白胆固醇(HDL-C)测定试剂盒,南京建成生物工程有限公司生产。
1.3 动物及分组
健康成年雄性SD大鼠,体重200±20g,每组8只;模型组饲料:在维持饲料中添加20.0%蔗糖、15%猪油、1.2%胆固醇、0.2%胆酸钠,适量的酪蛋白、磷酸氢钙、石粉等。除了粗脂肪外,模型饲料的水分、粗蛋白、粗脂肪、粗纤维、粗灰分、钙、磷、钙、磷均要达到维持饲料的国家标准。
实验分为空白对照组、模型对照组和3个剂量组的稠李提取物给药组,按体重随机分成2组,10只大鼠给予维持饲料作为空白对照组,40只给予模型饲料作为模型对照组。每周称量体重1次。模型对照组给予模型饲料1-2周后,空白对照组和模型对照组大鼠不禁食采血(眼内眦或尾部),采血后尽快分离血清,测定血清TC水平。根据TC水平将模型对照组随机分成4组,及模型组10只,3个剂量的给药组各10只。
1.4药物配置及给药方式
稠李提取物用生理盐水配置,按100, 200, 400mg/kg设为低、中、高三个剂量组,采用灌胃方式给药,空白对照组和模型对照组同时给予同体积的相应溶剂,空白对照组继续给予维持饲料,模型对照组及3个剂量组继续给予模型饲料,并定期称量体重。
1.5 血清学指标的测定
经4周给药后,麻醉动物,采血,采血后尽快分离血清,严格按试剂盒说明书方法测定血清TC、TG、LDL-C、HDL-C水平。
2 实验结果
经高脂饮食饲料喂养后,大鼠血脂TC, TG, LDL-C水平均明显高于空白对照组,且具有统计学意义(P<0.05);给予4周的稠李提取物后,中、高剂量(200,400mg/kg)均可以显著降低高血脂症大鼠的血脂水平(P<0.05);此外,中、高两个剂量组均可以显著升高血脂中HDL-C的水平(P<0.05)。
表1 稠李提取物对大鼠血清中TC、TG、LDL-C、HDL-C水平的影响(mmol/L)( ,n =8)
组别 | 剂量 | TG | TC | HDL-C | LDL-C |
空白对照组 | ---- | 0.89±0.23 | 1.76±0.45 | 0.93±0.19 | 0.71±0.27 |
模型组 | ---- | 1.22±0.32* | 2.79±0.56* | 0.71±0.21* | 1.88±0.43* |
稠李提取物低剂量组 | 100 | 1.19±0.49 | 2.66±0.49 | 0.72±0.23 | 1.86±0.39 |
稠李提取物中剂量组 | 200 | 1.11±0.36# | 2.28±0.42# | 0.82±0.19# | 1.77±0.36# |
稠李提取物高剂量组 | 400 | 1.02±0.28# | 1.87±0.39# | 0.88±0.22# | 1.65±0.37# |
“*”模型组与空白对照组相比较,“#”给药组与模型组相比较,差异显著(P<0.05)。
研究表明:高脂饮食会导致体内的抗氧化能力损伤,结果表明:高脂饮食饲养后,大鼠血清中SOD活力下降,MDA水平升高,经给药治疗后,稠李提取物中、高剂量(200, 400mg/kg)均可以显著降低高血脂症大鼠血清中的MAD水平,升高SOD活力(P<0.05)。
表2 稠李提取物对大鼠血清SOD和MDA水平的影响(mmol/L)(,n =8)
组别 | 剂量 | SOD | MDA |
空白对照组 | ---- | 232.6±23.4 | 3.17±1.71 |
模型组 | ---- | 208.3±29.7* | 6.55±2.13* |
稠李提取物低剂量组 | 100 | 203.8±19.7 | 6.46±2.33 |
稠李提取物中剂量组 | 200 | 217.2±21.6# | 4.88±1.66# |
稠李提取物高剂量组 | 400 | 228.1±31.2# | 4.19±1.89# |
“*”模型组与空白对照组相比较,“#”给药组与模型组相比较,差异显著(P<0.05)。
实施例4. 药品及保健品制备的实施例
制备药剂的实施例一
桔梗提取物20.00g,药用淀粉适量,两者充分混匀,装胶囊,制成100粒胶囊,每粒重0.25g,含桔梗提取物200mg。
制备药剂的实施例二
桔梗提取物20.00g,以1,2-丙二醇1000ml溶解,过滤后分装于1000只安瓶中,每瓶1.0ml,含桔梗提取物20mg。
本发明除了上述实施方式之外,其它等同技术方案也应当在其保护范围之内,在此不再一一叙述 。
Claims (7)
1.一种稠李提取物的制备方法,具体包括如下步骤:干燥的稠李样品,粉碎过20目筛,经适当的溶剂和提取方法,得到提取液,减压浓缩至干;用水混悬至无沉淀,上大孔树脂吸附柱色谱,依次用水和70%以上的乙醇进行洗脱,收集醇洗脱部分,即得稠李提取物。
2.根据权利要求1所述的稠李,选自山桃稠李(Padus maackii),稠李(Padus racemosa)或紫叶稠李(Padus virginiana)的任何一种;提取部位选自其果实,茎叶或花等任一部位,其优选果实。
3.根据权利要求1所述的提取溶剂和提取方法,提取溶剂选自水,甲醇,乙醇任何一种及不同比例混合物;提取方法选自浸泡提取、回流提取、超声提取、逆流提取、超临界萃取等的任何一种。
4.根据权利要求1所述的大孔吸附柱色谱,主要包括药学上能够接受的,可以用于分离的多种常规树脂,选自D101,NKA12或ADS等的任何一种。
5.根据权利要求1所述的稠李提取物,具有降血脂作用,可以用于药物、保健品及食品添加剂中的应用。
6.根据权利要求1所述的稠李提取物,其含有丰富的花色苷,黄酮等成分。
7.根据权利要求5所述的用途,其特征在于所述药物或保健品片剂,胶囊剂,粉针剂,注射剂,丸剂,软胶囊,颗粒剂和贴剂等多种剂型。
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