CN113773907A - 一种不同部位香薷挥发油的制备及化学成分的检测 - Google Patents

一种不同部位香薷挥发油的制备及化学成分的检测 Download PDF

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CN113773907A
CN113773907A CN202110847542.2A CN202110847542A CN113773907A CN 113773907 A CN113773907 A CN 113773907A CN 202110847542 A CN202110847542 A CN 202110847542A CN 113773907 A CN113773907 A CN 113773907A
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周鸿立
朱孟楠
李雪
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Abstract

本发明公开了一种不同部位香薷挥发油化学成分的检测,包括以下步骤:采用水蒸气蒸馏法提取香薷挥发油,通过GC‑MS对香薷挥发油进行定性定量分析,得到其中香薷花成分相对含量较高的有BETA‑去氢香薷酮占53.34%、茎中含量较高的为1,1'‑[亚乙基双(氧基)]二丁烷69.36%、叶中含量较高的为BETA‑去氢香薷酮等物质约占8.11%,且在香薷的花、茎中均有4‑蒈烯成分,花和叶中均含有BETA‑去氢香薷酮物质。对香薷挥发油进行抑菌试验,得到香薷花挥发油对金黄色葡萄球菌有一定的抑制生长作用,香薷茎挥发油对枯草芽孢杆菌、短小芽孢杆菌和白色念珠菌均有一定的抑制生长作用,香薷叶挥发油对金黄色葡萄球菌、枯草芽孢杆菌、短小芽孢杆菌和白色念珠菌均有一定的抑制生长作用,对香薷地上部分挥发油进行抗氧化评价,其清除DPPH的IC50为9.72mg/mL。

Description

一种不同部位香薷挥发油的制备及化学成分的检测
技术领域
本发明属于植物资源综合利用技术领域,具体涉及一种不同部位香薷花、茎、叶挥发油化学成分与抑菌、抗氧化活性的比较。
背景技术
香薷是唇形科植物石香薷或江香薷的地上干燥部分,香薷的化学成分很多,以挥发油类、黄酮类、香豆素类等功能性成分为主,其中挥发油类成分已超百种,主要为单萜类、芳樟醇、月桂烯、酮类、杂环类等,也有其他特殊成分。香薷挥发油是由香薷中提出来的具有香气的挥发性油状混合物,为淡黄至褐黄色油状物,清香辛甜,可溶于石油醚、无水乙醇和乙酸乙酯等有机溶剂,难溶于水。挥发油系天然植物抗菌活性成分,具有持久的抗菌活性、药物残留少、与抗生素无配伍禁忌等优点,香薷挥发油具有良好的抗病毒、抑菌、抗氧化、抗炎等作用,用香薷的挥发油进行口腔抑菌试验,发现具有明显抑制菌斑的作用。用香薷挥发油做其它菌株的抑制试验,发现对金色葡萄球菌、脑膜炎双球菌、绿脓杆菌也有较强抑制作用。衰老是生命过程中的生物化学现象,主要是体内产生的自由基氧化细胞所致,香薷挥发油具有较强的抗氧化性,对DPPH自由基、超氧阴离子自由基、羟基自由基均有显著的清除效果,是一种天然的抗氧化剂。
吴婷婷、朱冬青等人在GC-MS结合保留指数分析紫花香薷不同部位挥发油的化学成分文章中提到,不同器官(花、茎、叶部)挥发油的主要组分亦有差异,茎和叶的挥发油化学成分组成基本一致,但各成分含量上差别很大,花中挥发油鉴定出 39 种组分,其均由萜类物质所组成,香薷茎中挥发油成分以脂肪族化合物为主,据研究从香薷花、茎、叶部的挥发油中共鉴定了 82 种化合物。茎和叶的油中共同成分有 12 种,叶和花的油中共同成分有 17 种,茎和花的油中相同成分有 9 种,但各成分含量上差别很大,花部与茎和叶中挥发油的组成存在差异,尤以叶的差别最大。茎中芳香成分含量较少,考察不同器官的挥发油成分变化,说明即使是同一种物种的不同器官,可能由于代谢或功能上的差异,挥发油成分亦略有差异。目前对中药香薷花、茎、叶中挥发油有待于更深一步的研究探索。申请专利将对北香薷(Mosla Chinensis Maxim)的花、茎、叶进行挥发油提取,开展化学成分解析及抑菌和抗氧化作用的研究。随着挥发油化学成分和有效部位更详细的研究,将对开发北香薷具有一定的现实意义。
发明内容
本发明在于提供一种不同部位北香薷挥发油化学成分与抑菌、抗氧化活性的比较,以解决上述背景中出现的问题。
为实现上述目的,本实验提供如下技术方案。
具体实施方式
下面的实施例是对本发明的进一步描述。
表格说明
表1为香薷花挥发油GC-MS分析结果;
表2为香薷茎挥发油GC-MS分析结果;
表3为香薷叶挥发油GC-MS分析结果;
表4为香薷花、茎、叶挥发油的抗微生物效果结果。
实施例一香薷挥发油的提取
利用水蒸气蒸馏法提取香薷挥发油,分别精密称取20g干燥的香薷花、茎、叶粉碎60-80目,置于1000ml烧瓶内,加入没过粉末的蒸馏水,连接水蒸气蒸馏装置后开始提取2-4h,直至不再有油产生,收集馏出液,向馏出液中加入体积比1:2-3二氯甲烷萃取,静置分层,取下层液体,35-40℃旋转蒸发至有机溶剂全部回收,香薷挥发油挂壁时,收集挥发油,置于冰箱内备用。
实施例二香薷挥发油GC-MS分析
取香薷花、茎、叶三个部位挥发油各1 μl(乙酸乙酯溶解),采用 GC-MS 仪进行分析。色谱条件:色谱柱:HP-5MS 5% Phenyl Methyl Siloxane (30 m×0.25 mm ,0.25 μm);载气:高纯 He,载气流速:1.0ml·min-1,分流比 20 ∶ 1;升温程序:柱起始温度45℃,保持 2 min,以 5℃·min-1升温至 300℃,保持2 min;进样口温度:250℃,进样量:1 μl。质谱条件:离子源 EI 源,电子能量 70 eV,离子源温度:280 ℃,四级杆温度:150 ℃,溶剂延迟:5.0 min,质量范围50~550 amu,扫描方式:全扫描。其中花成分相对含量较高的有BETA-去氢香薷酮占53.34%、茎中含量较高的为1,1'-[亚乙基双(氧基)]二丁烷69.36 %、叶中含量较高的为BETA-去氢香薷酮等物质约占8.11%,且在香薷的花、茎中均有4-蒈烯成分,花和叶中均含有BETA-去氢香薷酮物质。
表1香薷花挥发油GC-MS分析结果
Figure 976196DEST_PATH_IMAGE001
表2香薷茎挥发油GC-MS分析结果
Figure 432585DEST_PATH_IMAGE003
表3香薷叶挥发油GC-MS分析结果
Figure 984789DEST_PATH_IMAGE004
实施例三香薷挥发油的抗微生物作用
配制相应菌种(大肠杆菌、金黄色葡萄球菌、白色念珠菌、酵母菌、短小芽孢杆菌、枯草芽孢杆菌)的培养基和相应培养基的琼脂培养溶液,包扎灭菌,拿出大板,洗干净用酒精擦拭,放在操作台用紫外灯照射30min,先倒入琼脂溶液,冷却后再倒入培养基与菌悬液的混合液,将样品纸片放上去,培养24h,观察抑菌效果,实验所用菌原液,配置相应斜面培养基,在无菌操作台用接种签蘸原菌,旋转接种签把原菌均匀涂抹在斜面培养基表面,包扎培养(酵母,白念29℃,其他菌种37℃)24小时。取出斜面培养基,在无菌操作台,用移液枪共吸取10ml无菌水冲洗斜面,即制成菌悬液,倒入无菌锥形瓶即可,得到香薷花挥发油对金黄色葡萄球菌有一定的抑制生长作用,香薷茎挥发油对枯草芽孢杆菌、短小芽孢杆菌和白色念珠菌均有一定的抑制生长作用,香薷叶挥发油对金黄色葡萄球菌、枯草芽孢杆菌、短小芽孢杆菌和白色念珠菌均有一定的抑制生长作用。
表4香薷花、茎、叶挥发油的抗微生物效果结果
Figure 328177DEST_PATH_IMAGE005
实施例四香薷地上部分挥发油的抗氧化作用
精密称取0.0039g DPPH于100mL容量瓶中,加入无水乙醇定容至100mL,将香薷挥发油用无水乙醇溶解稀释至1、1.5、2.5、3.5、4.5mg/mL5个浓度,取样品2mL+等体积的0.1mmol/LDPPH溶液,混匀避光反应30min,在517nm出测吸光度为A1,取2mL样液+2mL蒸馏水在在517nm出测吸光度为A2,取2mLDPPH溶液+2mL乙醇在517nm出测吸光度为A0,再取2mL无水乙醇+2mL蒸馏水进行空白调零,得出DPPH自由基清除率IC50为9.72mg/mL,DPPH自由基清除率(%)=
Figure 878107DEST_PATH_IMAGE006
本研究水蒸气蒸馏法提取北香薷的不同部位(花、茎、叶)挥发油,通过GC-MS对香薷挥发油进行定性定量分析,得到其中花成分相对含量较高的有BETA-去氢香薷酮占53.34%、茎中含量较高的为1,1'-[亚乙基双(氧基)]二丁烷69.36 %、叶中含量较高的为BETA-去氢香薷酮等物质约占8.11%,且在香薷的花、茎中均有4-蒈烯成分,花和叶中均含有BETA-去氢香薷酮物质,对香薷挥发油进行抑菌试验,得到香薷花挥发油对金黄色葡萄球菌有一定的抑制生长作用,香薷茎挥发油对枯草芽孢杆菌、短小芽孢杆菌和白色念珠菌均有一定的抑制生长作用,香薷叶挥发油对金黄色葡萄球菌、枯草芽孢杆菌、短小芽孢杆菌和白色念珠菌均有一定的抑制生长作用,综上所述香薷花和叶均对金黄色葡萄球菌有抑制作用,且花只对金黄色葡萄球菌有抑制作用,说明BETA-去氢香薷酮物质具有抑制金黄色葡萄球菌的作用,对香薷地上部分挥发油进行抗氧化评价,其清除DPPH的IC50为9.72mg/mL。表明香薷挥发油具有一定的抑菌抗氧化效果,对未来进一步研究开发北香薷提供参考。

Claims (6)

1.一种不同部位香薷挥发油的制备及化学成分的检测,其特征在于采用水蒸气蒸馏法提取香薷挥发油,并对香薷的花、茎、叶进行GCMS化学成分分析、抑菌及抗氧化实验。
2.如权利要求1所述的一种不同部位香薷挥发油的制备及化学成分的检测,特征在于:所述的香薷挥发油是香薷的花、茎、叶三个不同部位提取。
3.如权利要求1所述的一种不同部位香薷挥发油的制备及化学成分的检测,特征在于:所述的香薷挥发油是水蒸气蒸馏法提取2-4h,1:2-3的二氯甲烷萃取,35-40℃旋转蒸发所制得。
4.如权利要求1所述的一种不同部位香薷挥发油的制备及化学成分的检测,特征在于:所述的香薷挥发油中其中花成分相对含量较高的有BETA-去氢香薷酮占53.34%、茎中含量较高的为1,1'-[亚乙基双(氧基)]二丁烷69.36 %、叶中含量较高的为BETA-去氢香薷酮等物质约占8.11%,且在香薷的花、茎中均有4-蒈烯成分,花和叶中均含有BETA-去氢香薷酮物质。
5.如权利要求1所述的一种不同部位香薷挥发油的制备及化学成分的检测,特征在于:所述的香薷花挥发油浓度为0.02g/mL对金黄色葡萄球菌有一定的抑制生长作用,香薷茎挥发油浓度为0.02g/mL对枯草芽孢杆菌、短小念珠菌和白色念珠菌均有一定的抑制生长作用,香薷叶挥发油浓度为0.02g/mL对金黄色葡萄球菌、枯草芽孢杆菌、短小念珠菌和白色念珠菌均有一定的抑制生长作用。
6.如权利要求1所述的一种不同部位香薷挥发油的制备及化学成分的检测,特征在于:所述的香薷地上部分挥发油具有一定的清除DPPH自由基的能力,IC50为9.72mg/mL。
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