宁夏地区大籽蒿精油成分分析及其抑菌性研究

doi:10.11924/j.issn.1000-6850.casb20190900663

摘要/Abstract

摘要：

为探究宁夏大籽蒿(Artemisia sieversiana Ehrhart ex Willd.)精油成分、抑菌功能及安全性,采用水蒸气蒸馏法进行精油提取,通过气相色谱-质谱连用法(GC-MS)进行成分分析,采用滤纸片法测定其对3种菌的抑菌圈直径,测定最低抑菌浓度(MIC)、最低杀菌浓度(MBC),并通过cck-8法检测大籽蒿精油对于2种皮肤细胞的毒性作用。结果表明,宁夏大籽蒿精油的主要成分为母菊薁(29.61%),樟脑(4.80%)、桉叶油醇(4.32%)、异木香酸甲酯(4.02%);精油对铜绿假单胞菌和大肠杆菌的最低抑菌浓度为0.0781 μg/mL,最低杀菌浓度为0.3125 μg/mL;金黄色葡萄球菌MIC为0.0195 μg/mL,MBC为0.078125 μg/mL;当精油浓度小于100 μg/mL时对角质细胞(HaCaT)和皮肤成纤维细胞(NHSF)均无毒性。综上,宁夏地区大籽蒿精油具有较强的抑菌活性且安全性较高,以上结果为宁夏大籽蒿精油开发提供理论依据,以期为当地脱贫致富提供科技支撑。

关键词: 大籽蒿精油, 成分分析, 抑菌性, 细胞毒性

Abstract:

To explore the chemical composition, antibacterial function and safety of the essential oil of Artemisia sieversiana Ehrhart ex Willd. in Ningxia, steam distillation was adopted to extract the essential oil, gas chromatography-mass spectrometry (GC-MS) was adopted for component analysis. The inhibition zone diameter of three strains was determined by filter paper method, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. CCK-8 method was used to detect the toxic effects of Artemisia sieversiana essential oil on skin cells. The results showed that the main components of Ningxia Artemisia sieversiana essential oil were chamazulene (29.61%), camphor (4.80%), eucalyptol (4.32%), methyl isocostate (4.02%). The minimum inhibitory concentration for Pseudomonas aeruginosa and Escherichia coli was 0.0781 μg/mL, the minimum bactericidal concentration was 0.3125 μg/mL. The MIC of Staphylococcus aureus was 0.0195 μg/mL, the MBC was 0.078125 μg/mL. When the concentration of essential oil was less than 100 μg/mL, there was no toxicity to keratinocytes (HaCaT) and skin fibroblasts (NHSF). In summary, the essential oil of Artemisia sieversiana in Ningxia has strong antibacterial activity and high safety. The above results can provide a theoretical basis for the development of essential oil of Artemisia sieversiana, and science and technology support for local poverty alleviation.

Key words: essential oil of Artemisia sieversiana, component analysis, antibacterial activity, cyto-toxicity

中图分类号:

张宸, 王文翠, 白璐, 师淑莲, 姚雷. 宁夏地区大籽蒿精油成分分析及其抑菌性研究[J]. 中国农学通报, 2020, 36(29): 62-68.

Zhang Chen, Wang Wencui, Bai Lu, Shi Shulian, Yao Lei. Chemical Composition and Antimicrobial Activity of the Essential Oil of Artemisia Sieversiana in Ningxia[J]. Chinese Agricultural Science Bulletin, 2020, 36(29): 62-68.

图/表 5

参考文献 30

[1]	Liu S J, Liao Z X, Tang Z S, et al. Phytochemicals and biological activities of Artemisia sieversiana[J]. Phyto-chemistry Reviews, 2016,16(3):441-460.
[2]	中国科学院西北高原生物研究所. 藏药志[M]. 西宁: 青海人民出版社, 1991: 33-34.
[3]	李洪玲, 张爱东, 青格乐, 等. 大籽蒿研究利用现状及展望[J]. 畜牧与饲料科学, 2014(1):52-54.
[4]	徐福春, 土旦卓玛, 桑吉, 等. 藏药大籽蒿提取物体外抗菌活性研究[J]. 西藏大学学报, 2013(4):46-49.
[5]	韩小冰. 蒿属植物粗提物杀虫和抑菌活性的初步研究[D]. 哈尔滨:东北林业大学, 2008.
[6]	Hailiang L I, Haikui C, Fuli X U, et al. Chemical Composition, Antimicrobial and Antioxidant Activities of the Essential Oil of Artemisia sieversiana[J]. Food Science, 2016,37(20):63-68.
[7]	Zhigzhitzhapova S V, Popov D V, Pintaeva E T, et al. Essential Oil from Artemisia sieversiana Willd. and Development of Related Oil-in-Water Emulsions[J]. Pharmaceutical Chemistry Journal, 2017,51(5):388-390.
[8]	刘群, 邓治邦. 大籽蒿挥发油化学成分的研究[J]. 东北师大学报:自然科学, 1991(2):63-66.
[9]	李国明, 张丽萍, 胡永亮, 等. GC-MS法分析瑞丽柠檬香茅草精油化学成分[J]. 热带农业科学, 2018,38(2):87-92.
[10]	王书平, 吴涛, 陆玉建. 几种芳香植物精油的抑菌活性研究[J]. 安徽农学通报, 2018,24(2):20-24.
[11]	顾仁勇, 刘莹莹. 山苍子精油抑菌及抗氧化作用的研究[J]. 食品科学, 2006,27(11):86-89.
[12]	Avar S, Maksimovi M, Vidic D, et al. Chemical composition and antioxidant and antimicrobial activity of es-sential oil of Artemisia annua L. from Bosnia[J]. Industrial Crops and Products, 2012,37(1):479-485.
[13]	姜黎, 王向未, 江英. 薰衣草精油抑菌作用的研究[J]. 农产品加工(学刊), 2011(4):73-75.
[14]	谭龙飞, 钟彩莲, 陈颖姬, 等. 咖喱粉精油的抗菌作用研究及GC/MS分析[J]. 中国调味品, 2003(11):6-10.
[15]	刘玉民, 刘亚敏, 李鹏霞. 枫香叶精油抑菌活性及抗氧化活性研究[J]. 食品科学, 2009,30(11):134-137.
[16]	Bezić Nada, Skocibusić Mirjana, Dunkić Valerija, et al. Composition and Antimicrobial Activity of Achillea clavennae L. Essential Oil[J]. Phytotherapy research, 2003,17(9):1037-1040. doi: 10.1002/ptr.1290 URL pmid: 14595583
[17]	Martins A P, Salgueiro L R, Gonçalves M J, et al. Essential oil composition and antimicrobial activity of Santiria trimera bark.[J]. Planta Medica, 2003,69(01):77-79.
[18]	陈小义, 徐瑞成, 陈莉, 等. 蟾蜍灵对人胃癌细胞系MGc-803的细胞毒作用[J]. 中草药, 2000,31(12):920-922.
[19]	冯华龙, 何升华, 黄飞强, 等. CCK-8法定量检测亚甲蓝对人髓核细胞的毒性[J]. 中国组织工程研究, 2018,22(16).
[20]	施钧慧, 汪聪慧. 香料质谱图集[M]. 北京:中国质谱学会, 1992: 18-221.
[21]	Ornano L, Venditti A, Ballero M, et al. Chemopreventive and Antioxidant Activity of the Chamazulene-Rich Essential Oil Obtained from Artemisia arborescens L. Growing on the Isle of La Maddalena, Sardinia, Italy[J]. Chemistry & Biodiversity, 2013,10(8):1464-1474. URL pmid: 23939794
[22]	Safayhi H, Sabieraj H, Sailer J, et al. Chamazulene: an antioxidant-type inhibitor of leukotriene B4 formation[J]. Planta Medica, 1994,60(05):410-413.
[23]	丁元刚, 马红梅, 张伯礼. 樟脑药理毒理研究回顾及安全性研究展望[J]. 中国药物警戒, 2012(1):38-42.
[24]	Marino M, Bersani C, Comi G. Impedance measurements to study the antimicrobial activity of essential oils from Lamiacea and Compositae[J]. International Journal of Food Microbiology, 2001,67(3):187-195. URL pmid: 11518428
[25]	Burt S A. Essential oils: Their antibacterial properties and potential applications in foods - A review[J]. Interna-tional Journal of Food Microbiology, 2004,4(3):233-253.
[26]	胡林峰, 许明录, 朱红霞. 植物精油抑菌活性研究进展[J]. 天然产物研究与开发, 2011,23(2):384-391.
[27]	黄丽沄, 马海旭, 郭阿君. 罗勒精油成分鉴定及抑菌活性测定[J]. 北华大学学报:自然科学版, 2018,19(02):257-261.
[28]	Marcello Militello, Alessandra Carrubba, María Amparo Blázquez. Artemisia arborescens L.: essential oil composition and effects of plant growth stage in some genotypes from Sicily. Journal of Essential Oil Research, 2012,24(3):229-235.
[29]	吕飞. 天然植物精油的抑菌活性及其作用机理研究[D]. 北京:北京化工大学, 2011.
[30]	王步江, 刘金福, 樊秀花, 等. 肉桂精油抑菌活性研究[J]. 食品与机械, 2011(6):166-167.

序号	英文名			中文名		分子式		CAS		相对百分含量/% （大于0.1%）
1	alpha-Pinene			α-蒎烯		C₁₀H₁₆		000080-56-8		0.1493
2	Camphene			莰烯		C₁₀H₁₆		000079-92-5		0.1359
3	Sabinene			桧烯		C₁₀H₁₆		003387-41-5		0.3013
4	beta-Myrcene			β-月桂烯		C₁₀H₁₆		000123-35-3		1.9872
5	2-Carene			2-蒈烯		C₁₀H₁₆		000554-61-0		0.2744
6	D-Limonene			D-柠檬烯		C₁₀H₁₆		000138-86-3		0.175
7	Eucalyptol			桉叶油醇		C₁₀H₁₈O		000470-82-6		4.3181
8	gamma-Terpinene			γ-萜品烯		C₁₀H₁₆		000099-85-4		0.6381
9	p-Cymene			对伞花烃		C₁₀H₁₄		000099-87-6		0.4136
10	1-Octen-3-ol			3-辛烯醇		C₈H₁₆O		003391-86-4		0.2952
11	Copaene			古巴烯		C₁₅H₂₄		003856-25-5		0.2007
12	Camphor			樟脑		C₁₀H₁₆O		000464-49-3		4.8012
13	(-)-beta-Bourbonene			β-波旁烯		C₁₅H₂₄		005208-59-3		0.3025
14	Linalool			芳樟醇		C₁₀H₁₈O		000078-70-6		2.1065
15	beta-Elemene			β-榄香烯		C₁₅H₂₄		000515-13-9		0.9462
16	(-)-Terpinen-4-ol			(-)-4-萜品醇		C₁₀H₁₈O		020126-76-5		2.8829
17	delta-Terpineol			δ-萜品醇		C₁₀H₁₈O		007299-42-5		0.1647
18	Humulene			α-律草烯		C₁₅H₂₄		006753-98-6		0.125
19	(E)-beta-Famesene			反式-β-金合欢烯		C₁₅H₂₄		018794-84-8		0.6374
20	alpha-Terpineol			α-萜品醇		C₁₀H₁₈O		000098-55-5		4.0113
21	Germacrene D			右旋大根香叶烯		C₁₅H₂₄		023986-74-5		2.7823
22	beta-Eudesmene			β-桉叶烯		C₁₅H₂₄		017066-67-0		0.709
23	alpha-Zingiberene			姜烯		C₁₅H₂₄		000495-60-3		0.2074
24	gamma-Elemene			γ-榄香烯		C₁₅H₂₄		029873-99-2		0.7043
25	alpha-Farnesene			α-金合欢烯		C₁₅H₂₄		000502-61-4		1.5835
26	Nerol			橙花醇		C₁₀H₁₈O		000106-25-2		0.2623
27	Geraniol			香叶醇		C₁₀H₁₈O		000106-24-1		0.1215
28	Butanoic acid, 2-methyl-,3,7-dimethyl-2,6 -octadienyl este			2-甲基丁酸-3,7-二甲基-2,6-辛二烯酯		C₁₄H₂₂O₂		051117-19-2		3.4774
29	Jasmone			茉莉酮		C₁₁H₁₆O		000488-10-8		0.397
30	Caryophyllene oxide			石竹烯氧化物		C₁₅H₂₄O		001139-30-6		0.2902
31	Junenol			桧醇		C₁₀H₁₆O		000472-07-1		0.2474
32	Nerolidol			橙花叔醇		C₁₅H₂₆O		000142-50-7		0.12
33	3,6-Dihydrochamazulene			3,6-二氢母菊薁		C₁₄H₁₆		018454-88-1		3.6365
34	Eugenol			丁香酚		C₁₀H₁₂O₂		000097-53-0		0.3196
35	tau-Cadinol			t-杜松醇		C₁₅H₂₆O		150320-52-8		0.4145
36	alpha-Cadinol			α-杜松醇		C₁₅H₂₆O		000481-34-5		0.6411
37	Methyl isocostate			异木香酸甲酯		C₁₄H₂₄O₂		122365-00-8		4.0157
38	Chamazulene			母菊薁		C₁₄H₁₆		000529-05-5		29.6104
39	beta-Costol			β-木香醇		C₁₅H₂₄O		000515-20-8		0.4347
40		gamma-Costol	γ-木香醇		C₁₅H₂₄O		065018-14-6		0.769
41		Dehydrochamazulene	脱氢母菊薁		C₁₄H₁₆		321732-25-6		0.549
42		Phytol	植醇		C₂₀H₄₀O		000150-86-7		0.1829
43		Chrysantenyl 2-methuylbutanoate	2-甲基丁酸菊烯酯		C₁₁H₂₀O₂		053820-13-6		2.883

序号	英文名			中文名		分子式		CAS		相对百分含量/% （大于0.1%）
1	alpha-Pinene			α-蒎烯		C₁₀H₁₆		000080-56-8		0.1493
2	Camphene			莰烯		C₁₀H₁₆		000079-92-5		0.1359
3	Sabinene			桧烯		C₁₀H₁₆		003387-41-5		0.3013
4	beta-Myrcene			β-月桂烯		C₁₀H₁₆		000123-35-3		1.9872
5	2-Carene			2-蒈烯		C₁₀H₁₆		000554-61-0		0.2744
6	D-Limonene			D-柠檬烯		C₁₀H₁₆		000138-86-3		0.175
7	Eucalyptol			桉叶油醇		C₁₀H₁₈O		000470-82-6		4.3181
8	gamma-Terpinene			γ-萜品烯		C₁₀H₁₆		000099-85-4		0.6381
9	p-Cymene			对伞花烃		C₁₀H₁₄		000099-87-6		0.4136
10	1-Octen-3-ol			3-辛烯醇		C₈H₁₆O		003391-86-4		0.2952
11	Copaene			古巴烯		C₁₅H₂₄		003856-25-5		0.2007
12	Camphor			樟脑		C₁₀H₁₆O		000464-49-3		4.8012
13	(-)-beta-Bourbonene			β-波旁烯		C₁₅H₂₄		005208-59-3		0.3025
14	Linalool			芳樟醇		C₁₀H₁₈O		000078-70-6		2.1065
15	beta-Elemene			β-榄香烯		C₁₅H₂₄		000515-13-9		0.9462
16	(-)-Terpinen-4-ol			(-)-4-萜品醇		C₁₀H₁₈O		020126-76-5		2.8829
17	delta-Terpineol			δ-萜品醇		C₁₀H₁₈O		007299-42-5		0.1647
18	Humulene			α-律草烯		C₁₅H₂₄		006753-98-6		0.125
19	(E)-beta-Famesene			反式-β-金合欢烯		C₁₅H₂₄		018794-84-8		0.6374
20	alpha-Terpineol			α-萜品醇		C₁₀H₁₈O		000098-55-5		4.0113
21	Germacrene D			右旋大根香叶烯		C₁₅H₂₄		023986-74-5		2.7823
22	beta-Eudesmene			β-桉叶烯		C₁₅H₂₄		017066-67-0		0.709
23	alpha-Zingiberene			姜烯		C₁₅H₂₄		000495-60-3		0.2074
24	gamma-Elemene			γ-榄香烯		C₁₅H₂₄		029873-99-2		0.7043
25	alpha-Farnesene			α-金合欢烯		C₁₅H₂₄		000502-61-4		1.5835
26	Nerol			橙花醇		C₁₀H₁₈O		000106-25-2		0.2623
27	Geraniol			香叶醇		C₁₀H₁₈O		000106-24-1		0.1215
28	Butanoic acid, 2-methyl-,3,7-dimethyl-2,6 -octadienyl este			2-甲基丁酸-3,7-二甲基-2,6-辛二烯酯		C₁₄H₂₂O₂		051117-19-2		3.4774
29	Jasmone			茉莉酮		C₁₁H₁₆O		000488-10-8		0.397
30	Caryophyllene oxide			石竹烯氧化物		C₁₅H₂₄O		001139-30-6		0.2902
31	Junenol			桧醇		C₁₀H₁₆O		000472-07-1		0.2474
32	Nerolidol			橙花叔醇		C₁₅H₂₆O		000142-50-7		0.12
33	3,6-Dihydrochamazulene			3,6-二氢母菊薁		C₁₄H₁₆		018454-88-1		3.6365
34	Eugenol			丁香酚		C₁₀H₁₂O₂		000097-53-0		0.3196
35	tau-Cadinol			t-杜松醇		C₁₅H₂₆O		150320-52-8		0.4145
36	alpha-Cadinol			α-杜松醇		C₁₅H₂₆O		000481-34-5		0.6411
37	Methyl isocostate			异木香酸甲酯		C₁₄H₂₄O₂		122365-00-8		4.0157
38	Chamazulene			母菊薁		C₁₄H₁₆		000529-05-5		29.6104
39	beta-Costol			β-木香醇		C₁₅H₂₄O		000515-20-8		0.4347
40		gamma-Costol	γ-木香醇		C₁₅H₂₄O		065018-14-6		0.769
41		Dehydrochamazulene	脱氢母菊薁		C₁₄H₁₆		321732-25-6		0.549
42		Phytol	植醇		C₂₀H₄₀O		000150-86-7		0.1829
43		Chrysantenyl 2-methuylbutanoate	2-甲基丁酸菊烯酯		C₁₁H₂₀O₂		053820-13-6		2.883

菌种	抑菌圈直径/mm
菌种	100%精油	50%精油	25%精油	青霉素钠	丙酮
大肠杆菌	13.6±0.4	12.3±0.6	8.4±0.6	15.5±0.4	0
铜绿假单胞菌	14.5±0.3	8.3±0.4	7.2±0.3	10.5±0.3	0
金黄色葡萄球菌	16.4±0.4	13.6±0.2	10.8±0.6	13.7±0.5	0

菌种	抑菌圈直径/mm
菌种	100%精油	50%精油	25%精油	青霉素钠	丙酮
大肠杆菌	13.6±0.4	12.3±0.6	8.4±0.6	15.5±0.4	0
铜绿假单胞菌	14.5±0.3	8.3±0.4	7.2±0.3	10.5±0.3	0
金黄色葡萄球菌	16.4±0.4	13.6±0.2	10.8±0.6	13.7±0.5	0

菌种	精油浓度/(μg/mL)
菌种	20	5	1.25	0.3125	0.0781	0.0195	0.0048	0.0012	0.0003
大肠杆菌	-	-	-	-	-	﹢	﹢	﹢	﹢
铜绿假单胞菌	-	-	-	-	-	﹢	﹢	﹢	﹢
金黄色葡萄球菌	-	-	-	-	-	-	﹢	﹢	﹢