恩施产区湖北贝母乙酸乙酯组分挥发性成分与抗菌活性研究

doi:10.3969/j.issn.1004-1524.2021.06.14

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (6): 1088-1094.DOI: 10.3969/j.issn.1004-1524.2021.06.14

恩施产区湖北贝母乙酸乙酯组分挥发性成分与抗菌活性研究

李宇¹(), 刘翠君¹, 艾伦强¹, 方昕悦², 刘玉², 袁名远², 冉亚兰³, 何美军¹^,^*()

1.湖北省农业科学院中药材研究所,国家中药材产业技术体系恩施综合试验站,湖北恩施 445000
2.湖北民族大学生物科学与技术学院,湖北恩施 445000
3.恩施土家族苗族自治州农业农村局,湖北恩施 445000

收稿日期:2020-09-20 出版日期:2021-06-25 发布日期:2021-06-25
通讯作者: 何美军
作者简介:*何美军,E-mail: 840940513@qq.com
李宇(1990—),男,湖北恩施人,土家族,硕士,助理研究员,主要从事中药材天然产物研究。E-mail: 977956996@qq.com
基金资助:
湖北省重点研发计划(2020BCA059);湖北省农业科学院中药材研究所2019年青年科学基金(2019ZYCJJ02)

Study on volatile components and antibacterial activity of ethyl acetate component of Fritillaria hupehensis Hsiao et K. C. Hsia

LI Yu¹(), LIU Cuijun¹, AI Lunqiang¹, FANG Xinyue², LIU Yu², YUAN Mingyuan², RAN Yalan³, HE Meijun¹^,^*()

1. Institute of Chinese Herbal Medicines, Hubei Academy of Agricultural Sciences, National Traditional Chinese Medicine Industry Technology System Enshi Comprehensive Test Station, Enshi 445000, China
2. College of Biological Science and Technology, Hubei Minzu University, Enshi 445000, China
3. Agricultural and Rural Bureau of Enshi Tujia and Miao Autonomous Prefecture, Enshi 445000, China

Received:2020-09-20 Online:2021-06-25 Published:2021-06-25
Contact: HE Meijun

摘要/Abstract

摘要：

为了研究恩施产区湖北贝母乙酸乙酯组分的抗菌活性和挥发性成分,以恩施产区湖北贝母乙酸乙酯组分为研究对象,运用滤纸片法测试对11株供试菌的抑菌活性;采用微量肉汤稀释法测定对供试菌的MIC值;运用GC-MS鉴定挥发性成分;运用气相色谱外标法测定贝母素乙含量。研究结果显示:湖北贝母乙酸乙酯组分对苏云金芽孢杆菌、溶藻弧菌、鲍曼不动杆菌有中度敏感抑制活性,其MIC值分别为2.37、5.93、11.86 mg·mL^-1;湖北贝母乙酸乙酯组分的抑菌活性与贝母素乙的抑菌活性存在显著性差异,推测贝母素乙不是湖北贝母乙酸乙酯组分中主要发挥抑菌活性的成分;从乙酸乙酯组分中共鉴定出16个挥发性代谢产物,主要成分为脂肪烃(4.55%)、脂肪酸(24.07%)、含氮生物碱(23.02%);湖北贝母乙酸乙酯组分中贝母素乙的含量为(16.19±0.16) mg·g^-1,加标回收率为96.58%,相对标准偏差为3.98%。湖北贝母乙酸乙酯组分的优良抑菌活性和抑菌广谱性,为开发优质植物来源的新型天然抗生素提供了重要参考。

关键词: 恩施, 湖北贝母, 乙酸乙酯组分, 挥发性成分, 抑菌活性, 贝母素乙

Abstract:

To study the volatile components and antibacterial activity of ethyl acetate component of F. hupehensis, the ethyl acetate component of F. hupehensis in Enshi region was selected as the research object. The filter paper method was used to test the antimicrobial activity against 11 clinically resistant bacteria. Microbroth dilution method was used to determine the MIC value of the tested bacteria. The volatile components were identified by GC-MS and the content of peiminine was determined by gas chromatography. The results showed that: the ethyl acetate had medium inhibitory activity against Bacillus thuringiensis, Vibrio alginolyticus XSBZ14, and Acinetobacter baumannii ATCC 19606, and it’s MIC values was 2.37, 5.93 and 11.86 mg·mL^-1 respectively; the antibacterial activitys of ethyl acetate fraction of Hubei Fritillaria was significantly different from that of peiminine, which was proved that peiminine was not the main antibacterial agent in the ethyl acetate component of Fritillaria hupehensis. A total of 16 volatile metabolites were identified, including aliphatic hydrocarbons (4.55%), fatty acids (24.07%) and alkaloids (23.02%). The content of peiminine in the ethyl acetate fraction of F. hupehensis was (16.19±0.06) mg·g^-1, and the average recovery was 96.58%, the relaitve standard deviation was 3.98%. Excellent and broad spectrum antibacterial activity of ethyl acetate component of F. hupehensis provided an important reference for the development of new natural antibiotics from plant sources.

Key words: Enshi, Fritillaria hupehensis, ethyl acetate component, volatile component, antibacterial activity, peiminine

中图分类号:

李宇, 刘翠君, 艾伦强, 方昕悦, 刘玉, 袁名远, 冉亚兰, 何美军. 恩施产区湖北贝母乙酸乙酯组分挥发性成分与抗菌活性研究[J]. 浙江农业学报, 2021, 33(6): 1088-1094.

LI Yu, LIU Cuijun, AI Lunqiang, FANG Xinyue, LIU Yu, YUAN Mingyuan, RAN Yalan, HE Meijun. Study on volatile components and antibacterial activity of ethyl acetate component of Fritillaria hupehensis Hsiao et K. C. Hsia[J]. Acta Agriculturae Zhejiangensis, 2021, 33(6): 1088-1094.

图/表 4

参考文献 23

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序号 No.	挥发组分名称 Name of volatile components	保留时间 Retention time/min	化学式 Chemical formula	相对含量 Relative content/%	匹配度 Matching degree/%
1	2-甲基十六烷-1-醇 2-Methylhexadecan-1-ol	5.898	C₁₇H₃₆O	0.62	91.2
2	3,7,11-三甲基十二烷 3,7,11-Trimethyldodecan-1-ol	6.275	C₁₅H₃₂O	0.11	90.8
3	油酸, 3-丙酯 Oleic acid 3-(octadecyloxy)propyl ester	6.669	C₃₉H₇₆O₃	1.83	92.6
4	2-异丙基-5-甲基-1-庚醇 2-Isopropyl-5-methylheptan-1-ol	6.792	C₁₁H₂₄O	0.26	96.3
5	2,4-二乙基庚烷-1-醇 2,4-Diethylheptan-1-ol	7.045	C₁₁H₂₄O	0.79	96.2
6	苯甲酸乙基己酯 2-Ethylhexyl benzoate	8.575	C₁₅H₂₂O₂	1.25	90.5
7	棕榈酸甲酯 Hexadecanoic acid.methyl ester	9.992	C₁₇H₃₄O₂	3.82	96.3
8	棕榈酸乙酯 Ethyl palmitate	10.475	C₁₈H₃₆O₂	1.42	95.2
9	棕榈酸 Palmitic acid	10.604	C₁₆H₃₂O₂	3.92	97.2
10	13-表泪柏醇 13-Epimanool	11.374	C₂₀H₃₄O	1.92	98.7
11	亚油酸乙酯 Ethyl (9Z,12Z)-octadeca-9,12-dienoate	12.022	C₂₀H₃₆O₂	1.12	96.8
12	亚油酸 (9Z,12Z)-octadeca-9,12-dienoic acid	12.151	C₁₈H₃₂O₂	9.56	97.5
13	油酸 Oleic acid	12.310	C₁₈H₃₄O₂	3.52	92.1
14	1-羟基三十烷 Heptatriacontan-1-ol	14.110	C₃₇H₇₆O	0.85	92.8
15	1-O,3-O-二苄基6-O-甲基2-吲哚-1,3,6-三羧酸	21.715	C₂₈H₂₅NO₇	12.16	98.1
	1-O,3-O-dibenzyl 6-O-methyl 2-(methoxymethyl)indole-1,3,6-tricarboxylate
16	贝母素乙 Peiminine	23.4793	C₂₇H₄₃NO₃	10.86	91.8

序号 No.	挥发组分名称 Name of volatile components	保留时间 Retention time/min	化学式 Chemical formula	相对含量 Relative content/%	匹配度 Matching degree/%
1	2-甲基十六烷-1-醇 2-Methylhexadecan-1-ol	5.898	C₁₇H₃₆O	0.62	91.2
2	3,7,11-三甲基十二烷 3,7,11-Trimethyldodecan-1-ol	6.275	C₁₅H₃₂O	0.11	90.8
3	油酸, 3-丙酯 Oleic acid 3-(octadecyloxy)propyl ester	6.669	C₃₉H₇₆O₃	1.83	92.6
4	2-异丙基-5-甲基-1-庚醇 2-Isopropyl-5-methylheptan-1-ol	6.792	C₁₁H₂₄O	0.26	96.3
5	2,4-二乙基庚烷-1-醇 2,4-Diethylheptan-1-ol	7.045	C₁₁H₂₄O	0.79	96.2
6	苯甲酸乙基己酯 2-Ethylhexyl benzoate	8.575	C₁₅H₂₂O₂	1.25	90.5
7	棕榈酸甲酯 Hexadecanoic acid.methyl ester	9.992	C₁₇H₃₄O₂	3.82	96.3
8	棕榈酸乙酯 Ethyl palmitate	10.475	C₁₈H₃₆O₂	1.42	95.2
9	棕榈酸 Palmitic acid	10.604	C₁₆H₃₂O₂	3.92	97.2
10	13-表泪柏醇 13-Epimanool	11.374	C₂₀H₃₄O	1.92	98.7
11	亚油酸乙酯 Ethyl (9Z,12Z)-octadeca-9,12-dienoate	12.022	C₂₀H₃₆O₂	1.12	96.8
12	亚油酸 (9Z,12Z)-octadeca-9,12-dienoic acid	12.151	C₁₈H₃₂O₂	9.56	97.5
13	油酸 Oleic acid	12.310	C₁₈H₃₄O₂	3.52	92.1
14	1-羟基三十烷 Heptatriacontan-1-ol	14.110	C₃₇H₇₆O	0.85	92.8
15	1-O,3-O-二苄基6-O-甲基2-吲哚-1,3,6-三羧酸	21.715	C₂₈H₂₅NO₇	12.16	98.1
	1-O,3-O-dibenzyl 6-O-methyl 2-(methoxymethyl)indole-1,3,6-tricarboxylate
16	贝母素乙 Peiminine	23.4793	C₂₇H₄₃NO₃	10.86	91.8

生物碱 Alkaloid	检测浓度 Detection concentration/(mg·mL^-1)	回收率 Recovery rate/%	相对标准偏差 RSD/%	含量 Content/(mg·g^-1)
贝母素乙Peiminine	4.06	96.58	3.98	16.19±0.16
	4.06
	4.08
	4.02
	4.00
	4.02

生物碱 Alkaloid	检测浓度 Detection concentration/(mg·mL^-1)	回收率 Recovery rate/%	相对标准偏差 RSD/%	含量 Content/(mg·g^-1)
贝母素乙Peiminine	4.06	96.58	3.98	16.19±0.16
	4.06
	4.08
	4.02
	4.00
	4.02

供试菌种 Bacteria	乙酸乙酯组分 Ethyl acetate component/mm	贝母素乙 Peiminine/mm	MIC/ (mg·mL^-1)	抑菌效果 Antibacterial effect
肺炎克雷伯菌 Klebsiella pneumonia ATCC 13883	8.85±0.40	—^**	—	微敏Microsensitivity
金黄色葡萄球菌 Staphylococcus aureus ATCC 29213	8.17±0.44	7.81±0.49	—	微敏Microsensitivity
粪链球菌 Enterococcus faecalis ATCC 29212	8.02±0.38	—^**	—	微敏Microsensitivity
枯草芽孢杆菌 Bacillus subtilis	8.43±0.49	6.43±0.13^*	—	微敏Microsensitivity
藤黄微球菌 Micrococcus luteus	9.99±0.65	6.49±0.20^**	—	微敏Microsensitivity
苏云金芽胞杆菌 Bacillus thuringiensis	13.32±1.08	—^**	2.37	中敏Mild sensitivity
耐甲氧西林金黄色葡萄球菌 methicillin-resistant Staphylococcus aureus	7.87±0.52	—^**	—	微敏Microsensitivity
铜绿色假单胞菌 Pseudomonas aeruginosa	7.47±0.31	6.76±0.35	—	微敏Microsensitivity
鲍曼不动杆菌 Acinetobacter baumannii ATCC 19606	12.15±0.47	8.67±1.28^**	11.86	中敏Mild sensitivity
溶藻弧菌 Vibrio alginolyticus XSBZ14	12.18±0.88	6.60±0.32^**	5.93	中敏Mild sensitivity
表皮葡萄球菌 methicillin-resistant Staphylococcus epidermidis	7.34±0.52	7.42±0.61	—	微敏Microsensitivity

恩施产区湖北贝母乙酸乙酯组分挥发性成分与抗菌活性研究

Study on volatile components and antibacterial activity of ethyl acetate component of Fritillaria hupehensis Hsiao et K. C. Hsia

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