CN109568358A - 一种大叶补血草根中黄酮类和酚类化合物的用途 - Google Patents
一种大叶补血草根中黄酮类和酚类化合物的用途 Download PDFInfo
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Abstract
本发明涉及提供一种大叶补血草根中黄酮类和酚类化合物的用途,所述大叶补血草根中黄酮类和酚类化合物在制备清除自由基或抑制葡萄球菌中的用途,并对其进行了体外1,1‑二苯基‑三硝基苯肼(DPPH)清除自由基能力活性和抑制三种致病菌生长活性的测定,结果表明:没食子酸甲酯、杨梅素、二氢杨梅素、和没食子酸具有较好清除自由基活性,活性强度为:没食子酸>杨梅素>二氢杨梅素>没食子酸甲酯;杨梅素和槲皮素对葡萄球菌具有抑制生长作用,用于开发制备抗氧化,抑菌作用的药品或保健品。
Description
技术领域
本发明涉及医药技术领域,涉及一种大叶补血草根中黄酮类和酚类化合物的用途。
背景技术
大叶补血草(Limonium gmelinii),为白花丹科或蓝雪科(Plumbaginaceae)补血草属(Limonium Mill)多年生草本盐生植物,分布较广,是补血草中群落面积最大的一种,国内主要分布在中国新疆北部,在伊犁地区、塔城地区和阿勒泰地区均有分布,为维吾尔族和哈萨克族的传统用药。国外对大叶补血草研究主要是哈萨克斯坦阿尔法拉比国立大学化学学院的G.E Zhusupova和L.M.Korulkina教授等人,从该植物的根部分离获得半乳糖、五倍子酸、丁香酸、鞣花酸和几种黄酮类化合物及其苷类。其中根部含多种黄酮醇。大叶补血草具有清热祛湿,止血散瘀,消炎,治疗子宫内膜炎、宫颈糜烂、子宫出血、尿血痈疽等功效,另外有记载了其民族民间的6种附方。还未见大叶补血草中黄酮类化合物和酚类化合物的抗氧化、抗菌活性的报道。
黄酮类化合物具有较强的抗氧化作用,其抗氧化作用主要通过清除自由基发挥,现代科学研究工作者通过对多种植物类黄酮的研究表明均有较强的抗氧化作用,如中华补血草,银杏,槐角,豌豆根,双孢蘑菇等都具有较强的清除羟基自由基等抗氧化作用。
发明内容
本发明的目的在于,提供一种大叶补血草根中黄酮类和酚类化合物的用途,所述大叶补血草根中黄酮类和酚类化合物的在制备清除自由基或抑制葡萄球菌中的用途,并对其进行了体外1,1-二苯基-三硝基苯肼(DPPH)清除自由基能力活性和抑制三种致病菌生长活性的测定,结果表明:没食子酸甲酯、杨梅素、二氢杨梅素、和没食子酸具有较好清除自由基活性,活性强度为:没食子酸>杨梅素>二氢杨梅素>没食子酸甲酯;杨梅素和槲皮素对葡萄球菌具有抑制生长作用。
本发明所述的一种大叶补血草根中黄酮类和酚类化合物的用途,所述大叶补血草根中黄酮类和酚类化合物的在制备清除自由基药物中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和二氢杨梅素,酚类化合物没食子酸甲酯和没食子酸。
所述大叶补血草根中黄酮类化合物在制备抑制葡萄球菌的药物中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和槲皮素。
所述大叶补血草根中黄酮类和酚类化合物的在制备清除自由基保健品中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和二氢杨梅素,酚类化合物没食子酸甲酯和没食子酸。
所述大叶补血草根中黄酮类化合物在制备抑制葡萄球菌的保健品中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和槲皮素。
本发明所述的一种大叶补血草根中黄酮类和酚类化合物的用途,所述化合物是申请人2017年从大叶补血草根中分离鉴定的11个化合物,其结构式为:
具体操作按下列步骤进行:
a、将干燥的大叶补血草根20公斤粉碎,用浓度为95%的乙醇室温下冷浸提取3次,每次用2小时,料液比为:1:8,将提取液在温度45℃条件下用旋转蒸发仪减压蒸干,得到1.5公斤的提取物,浓缩至膏状;
b、将步骤a所得膏状依次采用石油醚,乙酸乙酯,氯仿,正丁醇进行萃取,通过旋转蒸发仪减压浓缩后得到各极性段的萃取液,石油醚相:232.4g,乙酸乙酯相:206.8g,氯仿相:158.24g,正丁醇相:876.3g,然后浓缩至膏状;
c、将步骤b所得的乙酸乙酯萃取物120g,上正向硅胶柱层析分离,用体积比100:0-1:1的氯仿-甲醇进行梯度洗脱,得到30个流分Fr1-Fr30,流分经硅胶薄层层析(TLC)分析,将流分Fr1-Fr8合并,得到合并流分F1;
d、将步骤c合并流分,F1上凝胶柱,用体积比1:1的氯仿-甲醇进行洗脱,分离得到二氢丁香亭和香草酸;
e、将流分F3上凝胶柱,用体积比1:1的氯仿-甲醇进行洗脱,得到小部位,对小流分F3-11进行薄层板制备(PTLC)纯化得到酪醇;对F3-21-23流分用制备液相色谱(PHPLC),制备得到柚皮素;
f、将流分F4上凝胶柱,用体积比1:1的氯仿-甲醇进行洗脱,得到了小部位,对小部位F4-15-19再次上凝胶柱,用体积比1:1的氯仿-甲醇进行洗脱,分离得到的F4-15-19-8-9流分,进行纯化得到香橙素;再将小流分F4-23-29进行纯化得到山奈酚;
g、将流分F6上反向硅胶(ODS)柱,依次用甲醇0%,10%,20%,30%,40%,50%,60%进行洗脱,其中浓度为30%甲醇洗脱得到没食子酸甲酯;
h、将流分F7上凝胶柱,用甲醇洗脱,得到若干小流分,对其中小部位F8-80-84进行纯化,得到槲皮素;
j、将流分F8上凝胶柱,用甲醇洗脱,得到若干小流分,对其中小部位F9-42-47,F9-62-63进行纯化,分离得到二氢杨梅素和为杨梅素;
k、将流分F9用快速分离色谱(Flash)纯化,对析出结晶的小部位F10-9-31进行纯化,得到没食子酸。
对于从大叶补血草中分离鉴定的11个具有类似结构的黄酮类和酚类化合物用1,1-二苯基-三硝基苯肼(DPPH)法测定抗氧化清除自由基和抑菌活性,结果显示:没食子酸甲酯、杨梅素、二氢杨梅素和没食子酸具有较好抗氧化清除自由基活性,活性强度为:没食子酸>杨梅素>二氢杨梅素>没食子酸甲酯;杨梅素和槲皮素对葡萄球菌具有抑制生长作用。
具体实施方式
实施例1
抗氧化清除自由基活性的测定:精密配制2mM 1,1-二苯基-三硝基苯肼(DPPH)溶液作为储备液,将样品都溶解于二甲基亚砜(DMSO)溶液,配制为50mg/mL或100mg/mL当做样品母液,将1,1-二苯基-三硝基苯肼(DPPH)储备液稀释10倍,即2×10-4mol/L浓度,测定515nm波长下的紫外吸光值(OD),并调整溶液浓度使其紫外吸光值(OD)值为0.7,得到1,1-二苯基-三硝基苯肼(DPPH)工作溶液;将样品母液逐级稀释配制成不同浓度梯度的样品溶液,以维生素C作为阳性对照;配制反应体系:取受试样品100μL,及二甲基亚砜(DMSO)溶液100μL,配制后充分混匀,在96孔板中温室避光反应30分钟后,用酶标仪测量各孔在515nm处紫外吸光值,平行测定3次,1,1-二苯基-三硝基苯肼(DPPH)清除率(%)=[(A0-A1)/A0]×100%,其中,A0为不加样时的吸光值,A1为加样组的吸光值;用IC50软件计算,最少3值,跨越50%,见表1;
表1清除自由基能力的IC50值
由表可知,没食子酸甲酯、杨梅素、二氢杨梅素和没食子酸具有显著清除自由基能力,具有抗氧化活性,活性强度为:没食子酸>杨梅素>二氢杨梅素>没食子酸甲酯。
实施例2
纸片法测定抑菌活性实验:融化琼脂培养基,待其温度降至46±0.5℃,加入已培养好的菌液,使试验菌悬液浓度为5×105cfu/mL-5×106cfu/mL,倒平皿15-20Ml/皿,放置20min使其凝固,用琼脂打孔器打孔,直径为5-6mm,4-5孔/皿,均匀分布,各样片中心之间相距25mm以上,与平板的周缘相距15mm以上,样品浓度为100mg/mL(100mM);每孔加样品溶液20μL,盖好平皿,置于温度37℃培养箱30-60min,使溶液完全被吸收,倒置培养16-18h,用游标卡尺测量抑菌圈的直径并记录,抑菌圈直径大于7mm者,判为有抑菌作用,见表2;
表2抑菌活性测定
由表可知,杨梅素和槲皮素具有显著抑制葡萄球菌生长的能力。
本发明所述的一种大叶补血草根中黄酮类和酚类化合物的用途,经测试结果表明:该黄酮类和酚类化合物中没食子酸甲酯、杨梅素、二氢杨梅素和没食子酸具有显著清除自由基能力,具有抗氧化活性;杨梅素和槲皮素具有显著抑制葡萄球菌生长的能力,可以开发利用具有为抗氧、抑菌活性的药品或保健品中的用途。
Claims (4)
1.一种大叶补血草根中黄酮类和酚类化合物的用途,其特征在于,所述大叶补血草根中黄酮类和酚类化合物在制备清除自由基药物中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和二氢杨梅素,酚类化合物没食子酸甲酯和没食子酸。
2.一种如权利要求1所述的用途,其特征在于,所述大叶补血草根中黄酮类化合物在制备抑制葡萄球菌的药物中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和槲皮素。
3.如权利要求1所述的用途,其特征在于,所述大叶补血草根中黄酮类和酚类化合物的在制备清除自由基保健品中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和二氢杨梅素,酚类化合物没食子酸甲酯和没食子酸。
4.一种如权利要求2所述的用途,其特征在于,所述大叶补血草根中黄酮类化合物在制备抑制葡萄球菌的保健品中的用途,其中大叶补血草根中黄酮类化合物为杨梅素和槲皮素。
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张莉静等: "杨梅素体内抗菌抗炎药效学研究", 《时珍国医国药》 * |
池晓会: "大叶补血草活性成分的研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
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