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Study on the Chemical Composition and Bioactivity of Two Species of Hyssopus Genus
Alternative Title两种神香草属植物的化学成分及其生物活性研究
Shomirzoeva Omina Ismoilovna
Subtype博士
Thesis Advisor阿吉艾克拜尔·艾萨
2018-06-02
Degree Grantor中国科学院大学
Place of Conferral北京
Degree Discipline有机化学
KeywordHyssopus Genus Hyssopus Seravshanicus Hyssopus Cuspidatus Chemical Composition Isolation And Purification Structure Elusidation Quantitative Determination Hplc-dad-esi-hrms/ms Bioactivity
Abstract

本论文对两种分别来源于塔吉克斯坦瓦尔佐布和中国新疆的神香草属植物Hyssopus seravshanicus和硬尖神香草(H. cuspidatus)中的化学成分及其生物活性进行了研究。利用硅胶、凝胶等柱层析和半制备型高效液相色谱等分离技术和手段,从H. seravshanicus的乙酸乙酯部位中分离得到18个化合物。通过质谱、核磁共振等波谱数据分析以及文献数据比对,将这些化合物鉴定为迷迭香酸甲酯(HS-1),迷迭香酸(HS-2),3-O-咖啡酰基奎宁酸甲酯(HS-3),地黄苷(HS-4),1-O-β-d-吡喃葡萄糖基-2-羟基-4-丙烯基苯(HS-5),丁香酚-O-β-d-吡喃葡萄糖苷(HS-6),1-羟基-2-O-β-d-吡喃葡萄糖基-4-丙烯基苯(HS-7),amburoside A(HS-8),deacyl martynoside(HS-9),芹菜素(HS-10),异槲皮素(HS-11),香叶木苷(HS-12),木樨草素(HS-13),南酸枣苷(HS-14),熊果酸(HS-15),齐墩果酸(HS-16),β-谷甾醇(HS-17),和samarcandin(HS-18)。其中,化合物HS-1,HS-3 ~ HS-9,HS-11和HS-14是H. seravshanicus中首次分离得到,化合物HS-1,HS-3 ~ HS-9,HS-11和HS-14是神香草属中首次发现。除了以上分离鉴定的18个化合物之外,该论文建立了超高压液相色谱方法,基于特征性紫外吸收对乙酸乙酯部位中的化合物类型进行了判定,发现酚类化合物是H. seravshanicus中的主要次生代谢产物,苯丙素类化合物尤其是丙烯酸衍生物是该植物中的代表性化合物。乙酸乙酯部位中的主要成分迷迭香酸甲酯(HS-1)的含量利用验证的UPLC方法确定为48.674 ± 0.714 mg/g。在抗氧化活性测试中,乙酸乙酯部位显示出一定的DPPH自由基清除能力和(IC50 = 19.15 ± 1.05 μg/ml)较强ATBS自由基清除能力(IC50 = 4.65 ± 0.33 μg/ml)。迷迭香酸甲酯(HS-1)表现出显著清除DPPH(IC50 = 5.22 ± 0.31 μM)和ATBS(IC50 = 3.71 ± 0.20 μM)自由基的能力,迷迭香酸(HS-2)(IC50 = 9.52 ± 0.66 μM),异槲皮素(HS-11)(IC50 = 13.55 ± 0.82 μM)和木樨草素(HS-13)(IC50 = 11.96 ± 0.60 μM)具有较强抗氧化活性,其 amburoside A(HS-8)展现出适当的抗氧化能力,而芹菜素(HS-10)只表现出弱的抗氧化能力。此外,乙酸乙酯部位、迷迭香酸甲酯(HS-1),amburoside A(HS-8)和异槲皮素(HS-11)还被发现可以提高小鼠B16黑色素瘤细胞中的黑色素含量,迷迭香酸甲酯(HS-1)和异槲皮素(HS-11)可以提高B16黑色素瘤细胞中的酪氨酸酶活性。从硬尖神香草乙酸乙酯部位中分离得到18个化合物,它们的结构通过质谱、核磁等波谱数据分析和文献数据对比得以确定。从H. seravshanicus中分离鉴定的五个化合物——迷迭香酸甲酯(HS-1),迷迭香酸(HS-2),3-O-咖啡酰基奎宁酸甲酯(HS-3),木樨草素(HS-13)和熊果酸(HS-15)也从硬尖神香草中分离得到。其余的13个化合物被鉴定为4-O-咖啡酰基奎宁酸甲酯(HC-1),咖啡酸(HC-3),七叶内酯(HC-4),三裂鼠尾草素(HC-5),芹菜素-4'-O-β-d-吡喃葡萄糖苷(HC-6),槲皮素-3-O-β-d-吡喃葡萄糖苷(HC-7),淫羊藿苷(HC-8),淫羊藿苷A(HC-9),朝藿定C(HC-10),glochidioboside [(7S,8R)- 二氢去氢二愈创木基醇-9'-O-β-d-吡喃葡萄糖苷(HC-11),胡萝卜甙(HC-12),和2α,3β,24-三羟基-12-烯-28-熊果酸(HC-13)。化合物HC-1,HC-2,HC-4,HC-6,HC-7 ~ HC-11和HC-14为神香草属植物中首次分离,迷迭香酸甲酯(HS-1)是从硬尖神香草中首次发现。与H. seravshanicus 类似,硬尖神香草中含量最高的化合物也是迭香酸甲酯(HS-1)和迷迭香酸(HS-2)。除了以上分离鉴定的18个化合物,硬尖神香草中的化学成分还利用Q-Exactive高分辨质谱仪,通过高分辨液质联用技术结合特征紫外吸收利用(HPLC-DAD-ESI-HRMS/MS)进行了分析,从中鉴定出9个酚类化合物。抗氧化活性实验表明,乙酸乙酯部位具有一定的抗氧化活性(IC50 = 26.53 ± 2.32 μg/ml),咖啡酸(HC-3)显示出显著抗氧化能力(IC50 = 24.68 ± 2.14 μM),oresbiusin A(HC-3)展现出一定抗氧化活性,而4-O-caffeoylquinic acid methyl ester(HC-1)则具有弱的抗氧化能力。抗菌活性测试发现,乙酸乙酯部位对白色念珠菌、大肠杆菌和金色葡萄球菌具有一定的抑制作用,迷迭香酸(HS-2)对白色念珠菌具有抑制作用,而oresbiusin A(HC-1)、三裂鼠尾草素(HC-5)和胡萝卜甙(HC-12)则对金色葡萄球菌具有抑制作用。

Other Abstract

Investigations on chemical composition of the whole plants of two Hyssopus plants, H. seravshanicus collected in Varzob region of Tajikistan and H. cuspidatus growing in Xinjiang, China, and their bioactivities were carried out.Eighteen compounds were obtained from the ethyl acetate fraction of H. seravshanicus by column chromatographies of silica gel and sephadex LH-20, as well as semi-preparative HPLC. On the basis of extensive spectroscopic data analysis including mass and NMR and comparison with previously reported data, the isolated compounds were identified to be methyl rosmarinate (HS-1), rosmarinic acid (HS-2), 3-O-caffeoylquinic acid methyl ester (HS-3), martynoside (HS-4), 1-O-β-d-glucopyranosyl-2-hydroxy-4-allylbenzene (HS-5), eugenyl-O-β-d-glucopyryranoside (HS-6), 1-hydroxyl-2-O-β-d-glucopyranosyl-4-allylbenzene (HS-7), amburoside A (HS-8), deacyl martynoside (HS-9), apigenin (HS-10), isoquercetin (HS-11), diosmin (HS-12), luteolin (HS-13), choerospondin (HS-14), ursolic acid (HS-15), oleanolic acid (HS-16), β-sitosterol (HS-17), and samarcandin (HS-18). Among them, compounds HS-1, HS-3 ~ HS-14 were discovered from H. seravshanicus for the first time, and compounds HS-1, HS-3 ~ HS-9, HS-11, and HS-14 were reported from the Hyssopus genus for the first time. Efforts on the diagnosis of the components except for the above isolated eighteen compounds according to the characteristic UV absorption have been attempted with a developed efficient UPLC method with good separation in a short analysis time. Phenolic compounds were found to be the main components of H. seravshanicus, and phenylpropoids, especially acrylic acid derivatives, were its representative constituents. The content of the most abundant compound, methyl rosmarinate (HS-1), in the ethyl acetate fraction of H. seravshanicus was determined to be 48.674 ± 0.714 mg/g with a validated UPLC method. In the DPPH radical scavenging assay, ethyl acetate fraction exhibited considerable DPPH radical scavenging ability (IC50 = 19.15 ± 1.05 μg/ml) and strong ATBS radical scavenging capacity (IC50 = 4.65 ± 0.33 μg/ml), methyl rosmarinate (HS-1) exhibited promising antioxidant activity in DPPH (IC50 = 5.22 ± 0.31 μM) and ABTS (IC50 = 3.71 ± 0.20 μM) assay, rosmarinic acid (HS-2) (IC50 = 9.52 ± 0.66 μM), isoquercetin (HS-11) (IC50 = 13.55 ± 0.82 μM) and HS-13 (IC50 = 11.96 ± 0.60 μM) showed strong antioxidant activity, amburoside A (HS-8) showed moderate antioxidant activity, while apigenin (HS-10) exhibited only slight antioxidant ability. Moreover, ethyl acetate fraction, methyl rosmarinate (HS-1), amburoside A (HS-8), and isoquercetin (HS-11) could increase the melanin content, and methyl rosmarinate (HS-1) and isoquercetin (HS-11) could also increase the activity of tyrosinase of B16 melanoma cells. From the ethyl acetate fraction of H. cuspidatus, eighteen compounds were isolated and their structures were determined with mass and NMR data investigations and comparison with literatures. Four compounds, methyl rosmarinate (HS-1), rosmarinic acid (HS-2), 3-O-caffeoylquinic acid methyl ester (HS-3), luteolin (HS-13), and ursolic acid (HS-15), which were discovered from H. seravshanicus, were also isolated and identified from H. cuspidatus. The other thirteen compounds were identified as 4-O-caffeoylquinic acid methyl ester (HC-1), oresbiusin A (HC-2), caffeic acid (HC-3), esculetin (HC-4), salvigenin (HC-5), apigenin-4'-O-β-d-glucopyranoside (HC-6), quercetin-3-O-β-d-glucopyranoside (HC-7), icariin (HC-8), epimedoside A (HC-9), epimedin C (HC-10), glochidioboside [(7S,8R)-dihydrodehydrodiconiferyl alcohol 9'-O-β-d-glucopyranoside] (HC-11), daucosterol (HC-12), and 2α,3β,24-trihydroxy-12-en-28-ursolic acid (HC-13). Methyl rosmarinate (HS-1) was revealed from H. cuspidatus for the first time, and compounds HC-1, HC-2, HC-4, HC-6, HC-7 ~ HC-11, and HC-13 were isolated from the Hyssopus genus for the first time. Similar to that of H. seravshanicus growing in Tajikistan, methyl rosmarinate (HS-1) and rosmarinic acid (HS-2) were found to be the most abundant compounds in the ethyl acetate fraction of H. cuspidatus. Besides above isolated eighteen compounds, further HPLC-DAD-ESI-HRMS/MS investigations by a high-resolution mass spectrometry with Q-Exactive mass spectrometer in combination with UV data analysis were applied to obtain more information of phenolic profile of H. cuspidatus. Nine phenolic compounds were identified by LC-MS/MS investigations. The ethyl acetate fraction (IC50 = 26.53 ± 2.32 mg/mL) of H. cuspidatus showed moderate antioxidant activity, caffeic acid (HC-3) (IC50 = 24.68 ± 2.14 μM) exhibited stronger scavenging capacity than that of the positive control, oresbiusin A (HC-2) (IC50 = 46.73 ± 3.23 μM) showed moderate antioxidant activity, and 4-O-caffeoylquinic acid methyl ester (HC-1) showed weak antioxidant activity. Ethyl acetate fraction and caffeic acid (HC-3) exhibited moderate inhibition ability to the growing of Candida albicans, Escherichia coli and Staphylococcus aureus, rosmarinic acid (HS-2) showed moderate antibacterial activity only against the fungus, C. albicans, while oresbiusin A (HC-2), salvigenin (HC-5), and daucosterol (HC-12) showed good antibacterial activity against gram positive bacteria, S. aureus.

Pages142
Document Type学位论文
Identifierhttp://ir.xjipc.cas.cn/handle/365002/5465
Collection资源化学研究室
Recommended Citation
GB/T 7714
Shomirzoeva Omina Ismoilovna. Study on the Chemical Composition and Bioactivity of Two Species of Hyssopus Genus[D]. 北京. 中国科学院大学,2018.
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