薄荷和留兰香资源挥发油抗菌活性和提取物抗氧化活性研究
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摘要
薄荷属植物(Mentha L.)为多年生或稀为一年生芳香草本,广泛分布于全球温带地区。薄荷属植物约30种,我国约12种,其中以薄荷Mentha haplocalyx Briq.和留兰香Mentha spicata L最为常见,应用最为广泛。薄荷和留兰香都是重要的药食兼用的植物,同时也是重要的工业原料。
为了更全面了解以四川省为主的不同来源地薄荷和留兰香资源挥发油组成和变异特点,检查其是否存在特殊的化学型和材料；揭示不同化学组成挥发油的抗菌活性差异；初步探讨不同来源薄荷和留兰香资源提取物抗氧化活性,为今后薄荷属植物的综合利用提供依据。本文以不同地区(主要为四川地区)收集到的薄荷和留兰香资源为对象,对其挥发油化学成分、抑菌活性和提取物抗氧化活性进行了研究,主要研究结果如下：
1.采用传统水蒸气蒸馏法,对收集的24份薄荷和留兰香地上部分挥发油进行提取,结合气相-质谱联用仪对挥发油进行分析鉴定分析。结果表明,所有材料挥发油组成和含量存在较大差异,在挥发油中共鉴定得到37个化学成分,主要成分为薄荷酮、薄荷醇、胡薄荷酮、香芹酮和氧化胡椒酮等。
2.根据挥发油主要化学成分及其相对百分含量,对24份薄荷和留兰香材料进行系统聚类分析,结果发现,所有材料可以聚为4类,即可以划分为四个化学型,包括薄荷酮-薄荷醇-胡薄荷酮型、香芹酮、薄荷醇型、薄荷酮型。所有留兰香材料均为香芹酮型；而薄荷材料则存在以上四种化学型。其中值得一提的是,材料M25和M26形态和薄荷M. haplocalyx接近,但化学成分组成、主要化学成分却和留兰香M. spicata一致,其化学型为香芹酮型。此外,四川薄荷(M.haplocalyx)资源中还首次发现薄荷酮型。
3.采用滤纸片扩散法和微量稀释法,以金黄色葡萄球菌、表皮葡萄球菌、大肠杆菌和白色念珠菌四种菌作为受试菌,分别测定了不同化学组成和含量的14份薄荷和留兰香材料挥发油抗菌活性和最小抑菌浓度(MIC),结果表明,材料主成分为薄荷酮、薄荷醇、胡薄荷酮的M31具有较强的抗菌作用；不同材料的抗菌活性强弱与主要化学成分有一定的关系,但是抗菌活性强弱并不完全取决于主要化学成分,说明其他含量较小的化学成分可能具有抗菌活性或者不同抗菌成分具有协同作用。
4.采用DPPH自由基清除法、ABTS自由基清除法和β-胡萝卜素漂白法对来自不同地区的薄荷和留兰香材料乙醇提取物进行抗氧化活性评价,结果显示,DPPH自由基清除法、ABTS自由基清除法测得的结果呈现一致性。其中材料M21和M26的自由基清除能力最强。β-胡萝卜素漂白法测定结果和DPPH法、ABTS法测得结果不尽一致,且试验发现所有材料抑制p-胡萝卜素漂白的能力均较弱。
5.采用分光光度法测定了薄荷和留兰香乙醇提取物的总酚和总黄酮含量。结果表明,材料M21和M26的总酚和总黄酮含量最高。相关分析结果表明,所有材料的自由基清除能力和总酚、总黄酮含量相关关系显著,抑制β-胡萝卜素漂白的能力与总酚、总黄酮含量相关不显著。
Genus Mentha L. accessions are perennial or far-between annual herbage, widely spread over the temperate zones of the world. The genus Mentha L. includes about 30 species worldwide, and about 12 species in our country. Among them, Mentha haplocalyx Briq. and Mentha spicata L. are two commom and widely used species. Both of them are used in food and medicine, meanwhile, they are also use as important industrial materials.
The purpose of this paper is to characterize the essential oil composi-tion and variations of M. haplocalyx and M. spicata from different areas (mainly from Sichuan province, China) and check whether there are specific chemotypes and materials in these areas, and reveal antimicrobial activity differences of essential oils with different composition, and also the antioxidant activity of mentha extracts, so as to provide reference for the future application of genus Mentha. This study used M. haplocalyx and M. spicata from different areas (mainly from Sichuan province, China) as the research materials. The composition and antimicrobial ac-tivity of the essential oils, and the antioxidant activity of extracts of M. haplocalyx and M. spicata were studied, and the main results were as follows:
1. To study the essential oils components of 24 M. haplocalyx and M. spicata collected in different areas, maily from Sichuan province, China, we extracted the volatile componentts by traditional hydro-distillation, and analyzed the components by GC-MS. The results showed that the constituents and conents of volatile oils in different Mentha resources were various. Totally,37 chemical components were identified from the volatile oils of 24 Mentha accessions. Menthone, menthol, pulegone, carvone, piperitone oxide etc. were found the main components.
2. Based on the percentage of the chemical components of the essen-tial oils of M. haplocalyx and M. spicata accessions,24 accessions were analyzed by cluster analysis. It was found that all the acces-sions were classified into 4 groups, namely 4 chemotypes, includ-ing menthone- menthol- pulegone, carvone, menthol, menthone. All the M.spicata accessions were chemotype carvone, the plant morphological characteristics of M25 and M26 were close to M. haplocalyx, but essential oils constituents, main components were in accordance with M. spicata, so its chemotype was carvone. The other M. haplocalyx accessions were divided into 4 different chemotypes. It was devoted to mention that the phenotype of M25 and M26 is similar to M. haplocalyx, but their chemotypes were the same as M. spicata, which belonged to chemotype carvone. Chemotype menthone was first report of M. haplocalyx collected in Sichuan province, China.
3. In this study,the antimicrobial activities of essential oils of 14 Mentha L. against Staphylococcus aureus ATCC29213, Staphylo-coccus epidermidis ATCC12228, Escherichia coli ATCC25922, Candida albicans ATCC 10231 were tested by the method of disc diffusion method and micro broth dilution. The results showed that M31 had strong inhibitory effects on all tested microorganisms, and its main components were menthone, menthol and pulegone. There are some relationships between antimicrobial activities and the major constituents of essential oils, but the antimicrobial ac-tivities were not completely depend on the main chemical compo-sitions of essential oils. It suggested there were some other com-pounds in the essential oils of Mentha L., which could display a series of physiological activities and multi-ingredient might have the synergistic effects.
4. The antioxidant activity of ethanol extracts of M. haplocalyx and M. spicata collected in different areas (maily from Sichuan province, China) was measured by three analytical methods:2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-di-(3-ethylbenzothialozine-sulphonic acid) (ABTS);β-carotene bleaching test.The results of DPPH·seavenging were consistent with ABTS+ scavenging. It has been shown by experi-mentation that the antioxidant activity of M21 and M26 was close to BHT. The results ofβ-carotene bleaching test were not consis-tent with the free radical scavenging activity, and the inhibitingβ-carotene bleaching activities of all the tested samples were rela-tively lower.
5. Total phenols, total flavonoids of ethanol extracts of M. haplocalyx and M. spicata were determined by spectrophotometry. As ex-pected, the phenolie content of M21 and M26 was much higher than other samples.The results of correlation analysis showed that there were significant relationships between the contents of total phenols, total flavonoids and the free radical scavenging activ-ity, while the contents of total phenols, total flavonoids exhibited no correlation with the inhibitingβ-carotene bleaching activities.
为了更全面了解以四川省为主的不同来源地薄荷和留兰香资源挥发油组成和变异特点,检查其是否存在特殊的化学型和材料；揭示不同化学组成挥发油的抗菌活性差异；初步探讨不同来源薄荷和留兰香资源提取物抗氧化活性,为今后薄荷属植物的综合利用提供依据。本文以不同地区(主要为四川地区)收集到的薄荷和留兰香资源为对象,对其挥发油化学成分、抑菌活性和提取物抗氧化活性进行了研究,主要研究结果如下：
1.采用传统水蒸气蒸馏法,对收集的24份薄荷和留兰香地上部分挥发油进行提取,结合气相-质谱联用仪对挥发油进行分析鉴定分析。结果表明,所有材料挥发油组成和含量存在较大差异,在挥发油中共鉴定得到37个化学成分,主要成分为薄荷酮、薄荷醇、胡薄荷酮、香芹酮和氧化胡椒酮等。
2.根据挥发油主要化学成分及其相对百分含量,对24份薄荷和留兰香材料进行系统聚类分析,结果发现,所有材料可以聚为4类,即可以划分为四个化学型,包括薄荷酮-薄荷醇-胡薄荷酮型、香芹酮、薄荷醇型、薄荷酮型。所有留兰香材料均为香芹酮型；而薄荷材料则存在以上四种化学型。其中值得一提的是,材料M25和M26形态和薄荷M. haplocalyx接近,但化学成分组成、主要化学成分却和留兰香M. spicata一致,其化学型为香芹酮型。此外,四川薄荷(M.haplocalyx)资源中还首次发现薄荷酮型。
3.采用滤纸片扩散法和微量稀释法,以金黄色葡萄球菌、表皮葡萄球菌、大肠杆菌和白色念珠菌四种菌作为受试菌,分别测定了不同化学组成和含量的14份薄荷和留兰香材料挥发油抗菌活性和最小抑菌浓度(MIC),结果表明,材料主成分为薄荷酮、薄荷醇、胡薄荷酮的M31具有较强的抗菌作用；不同材料的抗菌活性强弱与主要化学成分有一定的关系,但是抗菌活性强弱并不完全取决于主要化学成分,说明其他含量较小的化学成分可能具有抗菌活性或者不同抗菌成分具有协同作用。
4.采用DPPH自由基清除法、ABTS自由基清除法和β-胡萝卜素漂白法对来自不同地区的薄荷和留兰香材料乙醇提取物进行抗氧化活性评价,结果显示,DPPH自由基清除法、ABTS自由基清除法测得的结果呈现一致性。其中材料M21和M26的自由基清除能力最强。β-胡萝卜素漂白法测定结果和DPPH法、ABTS法测得结果不尽一致,且试验发现所有材料抑制p-胡萝卜素漂白的能力均较弱。
5.采用分光光度法测定了薄荷和留兰香乙醇提取物的总酚和总黄酮含量。结果表明,材料M21和M26的总酚和总黄酮含量最高。相关分析结果表明,所有材料的自由基清除能力和总酚、总黄酮含量相关关系显著,抑制β-胡萝卜素漂白的能力与总酚、总黄酮含量相关不显著。
Genus Mentha L. accessions are perennial or far-between annual herbage, widely spread over the temperate zones of the world. The genus Mentha L. includes about 30 species worldwide, and about 12 species in our country. Among them, Mentha haplocalyx Briq. and Mentha spicata L. are two commom and widely used species. Both of them are used in food and medicine, meanwhile, they are also use as important industrial materials.
The purpose of this paper is to characterize the essential oil composi-tion and variations of M. haplocalyx and M. spicata from different areas (mainly from Sichuan province, China) and check whether there are specific chemotypes and materials in these areas, and reveal antimicrobial activity differences of essential oils with different composition, and also the antioxidant activity of mentha extracts, so as to provide reference for the future application of genus Mentha. This study used M. haplocalyx and M. spicata from different areas (mainly from Sichuan province, China) as the research materials. The composition and antimicrobial ac-tivity of the essential oils, and the antioxidant activity of extracts of M. haplocalyx and M. spicata were studied, and the main results were as follows:
1. To study the essential oils components of 24 M. haplocalyx and M. spicata collected in different areas, maily from Sichuan province, China, we extracted the volatile componentts by traditional hydro-distillation, and analyzed the components by GC-MS. The results showed that the constituents and conents of volatile oils in different Mentha resources were various. Totally,37 chemical components were identified from the volatile oils of 24 Mentha accessions. Menthone, menthol, pulegone, carvone, piperitone oxide etc. were found the main components.
2. Based on the percentage of the chemical components of the essen-tial oils of M. haplocalyx and M. spicata accessions,24 accessions were analyzed by cluster analysis. It was found that all the acces-sions were classified into 4 groups, namely 4 chemotypes, includ-ing menthone- menthol- pulegone, carvone, menthol, menthone. All the M.spicata accessions were chemotype carvone, the plant morphological characteristics of M25 and M26 were close to M. haplocalyx, but essential oils constituents, main components were in accordance with M. spicata, so its chemotype was carvone. The other M. haplocalyx accessions were divided into 4 different chemotypes. It was devoted to mention that the phenotype of M25 and M26 is similar to M. haplocalyx, but their chemotypes were the same as M. spicata, which belonged to chemotype carvone. Chemotype menthone was first report of M. haplocalyx collected in Sichuan province, China.
3. In this study,the antimicrobial activities of essential oils of 14 Mentha L. against Staphylococcus aureus ATCC29213, Staphylo-coccus epidermidis ATCC12228, Escherichia coli ATCC25922, Candida albicans ATCC 10231 were tested by the method of disc diffusion method and micro broth dilution. The results showed that M31 had strong inhibitory effects on all tested microorganisms, and its main components were menthone, menthol and pulegone. There are some relationships between antimicrobial activities and the major constituents of essential oils, but the antimicrobial ac-tivities were not completely depend on the main chemical compo-sitions of essential oils. It suggested there were some other com-pounds in the essential oils of Mentha L., which could display a series of physiological activities and multi-ingredient might have the synergistic effects.
4. The antioxidant activity of ethanol extracts of M. haplocalyx and M. spicata collected in different areas (maily from Sichuan province, China) was measured by three analytical methods:2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-di-(3-ethylbenzothialozine-sulphonic acid) (ABTS);β-carotene bleaching test.The results of DPPH·seavenging were consistent with ABTS+ scavenging. It has been shown by experi-mentation that the antioxidant activity of M21 and M26 was close to BHT. The results ofβ-carotene bleaching test were not consis-tent with the free radical scavenging activity, and the inhibitingβ-carotene bleaching activities of all the tested samples were rela-tively lower.
5. Total phenols, total flavonoids of ethanol extracts of M. haplocalyx and M. spicata were determined by spectrophotometry. As ex-pected, the phenolie content of M21 and M26 was much higher than other samples.The results of correlation analysis showed that there were significant relationships between the contents of total phenols, total flavonoids and the free radical scavenging activ-ity, while the contents of total phenols, total flavonoids exhibited no correlation with the inhibitingβ-carotene bleaching activities.
引文
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