HPLC结合响应面法优化柳叶蜡梅总黄酮提取工艺及其抑菌活性研究

doi:10.3969/j.issn.1004-1524.2018.02.17

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (2): 298-306.DOI: 10.3969/j.issn.1004-1524.2018.02.17

HPLC结合响应面法优化柳叶蜡梅总黄酮提取工艺及其抑菌活性研究

温慧萍¹, 肖建中^{1, *}, 雷伟敏¹, 纪佳娜²

1.丽水学院生态学院,浙江丽水 323000;
2.丽水出入境检验检疫局综合技术服务中心,浙江丽水 323000

收稿日期:2017-11-20 出版日期:2018-02-20 发布日期:2018-02-11
通讯作者: 肖建中,E-mail:13905787007@163.com
作者简介:温慧萍(1983—),女,江西赣州人,硕士,助理研究员,从事中药的提取分析与药效研究。E-mail:happyxiaowen708@163.com
基金资助:
浙江省公益技术应用研究(分析测试类)项目(2016C37079)

Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.H by HPLC combined with response surface methodology and its antibacterial activity

WEN Huiping¹, XIAO Jianzhong^{1, *}, LEI Weimin¹, JI Jiana²

1. College of Ecology, Lishui University, Lishui 323000, China;
2. Technical Service Center, Lishui Entry-Exit Inspcction and Quarantine, Lishui 323000, China

Received:2017-11-20 Online:2018-02-20 Published:2018-02-11

摘要/Abstract

摘要： 采用高效液相色谱法测定柳叶蜡梅中黄酮类成分芦丁、槲皮素、山奈酚的含量,在单因素考察基础上,采用响应面法研究乙醇浓度、提取时间、提取温度对柳叶蜡梅黄酮类成分得率的影响,并采用滤纸片法对优化提取的柳叶蜡梅黄酮成分进行抑菌效果研究。结果表明,影响黄酮含量的因素为乙醇浓度>提取时间>提取温度;最佳提取工艺为提取温度90 ℃,料液比1:15,乙醇浓度79.0%,提取时间1.7 h;柳叶蜡梅中黄酮类成分为芦丁0.861 mg·g^-1,槲皮素0.504 mg·g^-1,山奈酚0.492 mg·g^-1,总提取量为1.857 mg·g^-1,与预测值的偏差为1.50%。优化提取的总黄酮对多种病原菌有抑制作用,对蜡样芽孢杆菌和大肠埃希菌有较强的抑制作用,抑菌圈直径分别为13.21、11.18 mm。可见,响应面优化柳叶蜡梅总黄酮提取工艺方法可行,提取的总黄酮具较好的抑菌效果,具有良好的开发前景。

关键词: 柳叶蜡梅, 黄酮, 响应面法, 抑菌作用

Abstract: The contents of rutin, quercetin and kaempferol extracted from Chimonanthus salicifolius were determined by high performance liquid chromatography (HPLC). Based on the single factor analysis, the effect of ethanol concentration, extraction time and extraction temperature on the yield of flavonoids from C. salicifolius were studied by response surface methodology. Meanwhile, the antibacterial activity of flavonoids from C. salicifolius was assessed by filter paper method. The results showed that main factors affecting the extraction of total flavonoids from C. salicifolius were ethanol concentration, extraction time, extraction temperature. The optimum extraction conditions of total flavonoids were: extraction temperature of 90 ℃, solid-liquid ratio of 1:15, ethanol concentration of 79%, extraction time of 1.7 h. Under the optimal conditions, contents of rutin, quercetin, kaempferol were 0.861, 0.504, 0.492 mg·g^-1, respectively; and the extraction yield of total flavonoids was 1.857 mg·g^-1 which differed from the theoretical value of 1.50%,the total flavonoids extracted from Chimonanthus salicifolius had antibacterial activity against many kinds of bacteria, especially Bacillus cereus and Escherichia coli, the mean diameters of the inhibition zones about them were 13.21 and 11.18 mm, respectively. Therefore, response surface methodology was feasible for the extraction optimization of total flavonoids from C. salicifolius. The total flavonoids from C. salicifolius had good antibacterial activity, indicating that it might have great application prospects in the fields of medicine.

Key words: Chimonanthus salicifolius, flavonoids, response surface methodology, antibacterial activity

中图分类号:

TQ461

温慧萍, 肖建中, 雷伟敏, 纪佳娜. HPLC结合响应面法优化柳叶蜡梅总黄酮提取工艺及其抑菌活性研究[J]. 浙江农业学报, 2018, 30(2): 298-306.

WEN Huiping, XIAO Jianzhong, LEI Weimin, JI Jiana. Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.H by HPLC combined with response surface methodology and its antibacterial activity[J]. , 2018, 30(2): 298-306.

参考文献

[1] GECHEV T S, HILLE J, WOERDENBAG H J, et al. Natural products from resurrection plants potential for medical applications[J]. Biotechnology Advances , 2014, 32(6): 1091-1101.
[2] LANZOTTI V. Drugs based on natural compounds: recent achievements and future perspectives[J]. Phytochemisty Reviews , 2014, 13(4): 725-726.
[3] FERREIRA A, POUSINHO S, FORTUNA A, et al. Flavonoid compounds as reversal agents of the P-glycoprotein-mediated multidrug resistance: biology, chemistry and pharmacology[J]. Phytochemisty Reviews , 2015, 14(2): 233-272.
[4] CAO J G, XIA X, CHEN X F, et al. Flavonoids contents, antioxidant and anticancer activities of 78 species of ferns from China[J]. Biomedical.Papers , 2012, 15(6): 33.
[5] PAVLOVA S I, ALBEGOVA D Z, VOROBEVA Y S, et al. Flavonoids as potential immunosuppressants affecting intracellular signaling pathways (a review)[J]. Pharmaceutical Chemistry Journal , 2016, 49(10): 645-652.
[6] LIU H T, CAO M P, ZHANG X F, et al. HPLC fingerprints establishment of chemical constituents in genus Chimonanthus leaves[J]. China Journal of Chinese Materia Medica , 2013, 38(16): 1560-1561.
[7] PAN X H, SHI X J, ZHANG X F, et al. Constituents of Chimonanthus salicifolius and their antioxidant activity[J]. Chinese Journal of Experimental Traditional Medical Formulas , 2012, 18(1): 99-102.
[8] WANG KW, DAN L, BIN W, et al. New cytotoxic dimeric and trimeric coumarins from Chimonanthus salicifolius [J]. Phytochemistry Letters , 2016, 16: 115-120.
[9] 刘忠达, 袁宙新, 张尊敬, 等. 柳叶蜡梅提取物对人胃癌SGC-7901细胞的作用机制研究[J]. 中华中医药杂志, 2011, 26(10): 2420-2423.
LIU Z D, YUAN Z X, ZHANG Z J, et al. Inhibitory effect of extract of Chimonanthus salicifolius on SGC-7901 cell and the possible mechanism[J]. Chinese Journal of Traditional Chinese Medicine , 2011, 26(10): 2420-2423. (in Chinese with English abstract)
[10] 王凌云, 张志斌, 邹峥嵘, 等. 蜡梅属植物化学成分和药理活性研究进展[J]. 时珍国医国药, 2012, 23(12): 3104-3106.
WANG L Y, ZHANG Z B, ZOU Z R, et al. Chemical constituents and pharmacological activities of the research progress of Chimonanthus [J]. Lishizhen Medicine and Materia Medica Research , 2012, 23(12): 3104-3106. (in Chinese with English abstract)
[11] 王宇翎, 张艳, 方明, 等. 白花蛇舌草总黄酮的抗炎及抗菌作用[J].中国药理学通报, 2005, 21(3): 348-350.
WANG Y L, ZHANG Y, FANG M, et al. Anti-inflammatory and antibacterial effects of total flavones of olden landia diffusa willd[J]. Chinese Pharmacological Bulletin , 2005, 21(3): 348-350. (in Chinese with English abstract)
[12] XIE R F,SHI Z N, LI Z C, et al. Optimization of high pressure machine decocting process for Dachengqi Tang using HPLC fingerprints combined with the Box-Behnken experimental design[J]. Journal of Pharmaceutical Analysis , 2015, 5(2): 110-119.
[13] SEVERINI C, DIEROSS A, FIORE A G. Ultrasound-assisted extraction to improve the recovery of phenols and antioxidants from spent espresso coffee ground: a study by response surface methodology and desirability approach[J]. European Food Research and Technology , 2017, 243(5): 835-847.
[14] ALEXANDRE E M C, ARAUJO P, FREITAS V D, et al. experimental design, modeling and optimization of high-pressure-assisted extraction of bioactive compounds from pomegranate peel[J]. Food and Bioprocess Technology , 2017, 10(5): 886-900.
[15] TARANTINO T B, BARBOSA I S, PERRIRA M D G, et al. Microwave-Assisted digestion using diluted nitric acid for multi-element determination in rice by ICP OES and ICP-MS[J]. Food Analytical Methods , 2017, 10(4): 1007-1015.
[16] 徐萌,张经纬, 张金凤. 响应面法优化柳叶蜡梅总黄酮叶的提取工艺[J]. 分子植物育种, 2016, 14(2): 496-500.
XU M, ZHANG J W, ZHANG J F. Optimizing extraction process of the flavonoids in Chimonanthus praecox leaves by response surface method[J]. Molecular Plant Breeding , 2016, 14(2): 496-500. (in Chinese with English abstract)
[17] MILUTINOVIC M, RADOVANOVIC N, COROVIC M, et al. Optimisation of microwave-assisted extraction parameters for antioxidants from waste Achillea millefolim dust[J]. Industrial Crops and Products , 2015, 77: 333-341.
[18] SNEZANA F, BRANINMINR P, SENKA V, et al. Optimization of microwave-assisted extraction of polyphenolic compounds from Ocimum basilicum by response surface methodology[J]. Food Analytical Methods , 2017, 10(7): 2270-2280.
[19] ZEKOVIC Z, VLADIC J, VLADIC S, et al. Optimization of microwave-assisted extraction (MAE) of coriander phenolic antioxidants-response surface methodology approach[J]. Journal of the Science of Food and Agriculture , 2016, 96(13): 4613-4622.
[20] CHENG Z Y, SONG H Y , CAO X L, et al. Simultaneous extraction and purification of polysaccharides from Gentiana scabra Bunge by microwave-assisted ethanol-salt aqueous two-phase system[J]. Industrial Crops and Products , 2017, 102: 75-87.
[21] JAERAL R, THAKUR S, SAKINAH M, et al. Potent anticancer, antioxidant and antibacterial activities of isolated flavonoids from Asplenium nidus [J]. Journal of King Saud University-Science , 2016. doi.org/10.1016/j.jksus.2016.11.006.
[22] WANG L J, YANG X S, QIN P Y, et al. Flavonoid composition, antibacterial and antioxidant properties of tartary buckwheat bran extract[J]. Industrial Crops and Products , 2013, 49(8): 312-317.
[23] AHMADU A A, ONANUGA A, EBESHI B U. Isolation of antibacterial flavonoids from the aerial parts of Indigofera secundiflora [J]. Pharmacognosy Journal , 2011, 3(19): 25-28.
[24] XIA X, CAO J G, ZHENG Y X, et al. Flavonoid concentrations and bioactivity of flavonoid extracts from 19 species of ferns from China[J]. Industrial Crops and Product s, 2014, 58(1): 91-98.
[25] JARIAL R, SHARD A, THAKUR S, et al. Characterization of flavonoids from fern Cheilanthes tenuifolia and evaluation of antioxidant, antimicrobial and anticancer activities[J]. Journal of King Saud University-Science , 2017. doi:doi.org/10.1016/j.jksus.2017.04.007.
[26] INAMULAH F, FATIMAL I, KHAN S, et al. New antimicrobial flavonoids and chalcone from Colutea armata [J]. Archives of Pharmacal Research , 2017, 40(8): 915-920.

HPLC结合响应面法优化柳叶蜡梅总黄酮提取工艺及其抑菌活性研究

Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.H by HPLC combined with response surface methodology and its antibacterial activity

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