混料设计优化孜然、花椒、芥子混合精油对4种果蔬采后真菌的抑菌效果
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摘要
本研究用水蒸馏法提取孜然、花椒、芥子精油,并对其成分作气相色谱-质谱联用分析;采用微量稀释法,琼脂纸片扩散法测定3种精油对4种果蔬真菌粉红单端孢(Trichothecium roseum)、扩展青霉(Penicillium expansum)、互隔交链孢(Alternaria alternata)、硫色镰刀菌(Fusarium sulphureum)的最小抑菌浓度(minimum inhibitory concentration,MIC)、最小杀菌浓度(minimum bactericidal concentration,MBC)和菌落直径,利用单纯型增强质心混料设计数学模型拟合最佳精油混料比例,并利用响应面和方差分析混和精油抑菌效果。结果表明提取得到的孜然精油、花椒精油、芥子精油含量分别为2.29、2.52、1.12 mL/g。孜然精油主要化学成分为枯茗醛(21.89%)、β-蒎烯(20.49%)、γ-萜品烯(18.37%)、4-丙-2-环己基-1,4-二烯-1-甲醛(10.98%)、p-伞花烃(8.89%)、4-丙-2-环己基-3-烯-1-甲醛(7.95%);花椒精油主要化学成分为芳樟醇(15.05%)、双戊烯(11.79%)、乙酸芳樟酯(11.17%)、β-蒎烯(9.33%)、(-)-4-萜品醇(6.04%)、桉叶油醇(4.62%);芥子精油的主要化学成分为烯丙基异硫氰酸酯(50.97%)、4-异硫氰基-1-丁烯(44.91%)。当孜然、花椒、芥子精油混料比例分别为36.11%、19.45%、44.44%时,对4种真菌的抑菌效果最佳,此时在统计学上存在显著性交互作用(P<0.05),具有良好的协同性;通过菌落直径大小分析得出混合精油相比于孜然精油、花椒精油单独作用具有较强的抑菌效果。该研究表明混料设计法可有效优化不同精油联合使用时的组合比,以实现较低精油用量下较好的抑菌效果,为天然抑菌剂的减量使用提供方法指导。
In this study, Cuminum cyminum L., Zanthoxylum bungeanum and Brassica juncea L. oils were extracted by steam distillation and their components were analyzed by gas chromatography-mass spectrometry. Using micro-dilution method and agar diffusion method, the minimum inhibitory concentration(MIC), minimum bactericidal concentration(MBC) and colony diameter of three essential oils against Trichothecium roseum, Penicillium expansum, Alternaria alternata and Fusarium sulphureum in fruits and vegetables were determined. The enhanced simple-centroid mixture design was used to establish a mathematical model to determine the best blend ratio of essential oils for higher antifungal effect employing response surface methodology and analysis of variance. The results showed that the extraction rates of essential oils from cumin, pricklyash and mustard were 2.29, 2.52 and 1.12 mL/g, respectively. The major chemical constituents in cumin oil were sebacic aldehyde(21.89%), β-pinene(20.49%), γ-terpinene(18.37%), 3-carene-10-aldehyde(10.98%),and p-cymene(8.89%). The major chemical constituents in pricklyash were linalool(15.05%), dipentene(11.79%), linalyl acetate(11.17%), β-pinene(9.33%) and(-)-4-terpineol(6.04%), and eucalyptol(4.62%). The major chemical components in mustard oil were allyl isothiocyanate(50.97%) and 4-isothiocyanato-1-butene(44.91%). The antibacterial effect of cumin, pricklyash and mustard essential oils when blended at a ratio of 36.11:19.45:44.44 was the best against each of the four fungi with significant interaction(P < 0.05) and good synergism being observed between the three oils. The results of this study demonstrated that mixture design could be useful to determine the optimum blend ratio of different essential oils for better antimicrobial effect at lower dosage, thereby providing methodological guidance for saving the dosage of natural bacteriostatic agents when used to preserve fruits and vegetables.
In this study, Cuminum cyminum L., Zanthoxylum bungeanum and Brassica juncea L. oils were extracted by steam distillation and their components were analyzed by gas chromatography-mass spectrometry. Using micro-dilution method and agar diffusion method, the minimum inhibitory concentration(MIC), minimum bactericidal concentration(MBC) and colony diameter of three essential oils against Trichothecium roseum, Penicillium expansum, Alternaria alternata and Fusarium sulphureum in fruits and vegetables were determined. The enhanced simple-centroid mixture design was used to establish a mathematical model to determine the best blend ratio of essential oils for higher antifungal effect employing response surface methodology and analysis of variance. The results showed that the extraction rates of essential oils from cumin, pricklyash and mustard were 2.29, 2.52 and 1.12 mL/g, respectively. The major chemical constituents in cumin oil were sebacic aldehyde(21.89%), β-pinene(20.49%), γ-terpinene(18.37%), 3-carene-10-aldehyde(10.98%),and p-cymene(8.89%). The major chemical constituents in pricklyash were linalool(15.05%), dipentene(11.79%), linalyl acetate(11.17%), β-pinene(9.33%) and(-)-4-terpineol(6.04%), and eucalyptol(4.62%). The major chemical components in mustard oil were allyl isothiocyanate(50.97%) and 4-isothiocyanato-1-butene(44.91%). The antibacterial effect of cumin, pricklyash and mustard essential oils when blended at a ratio of 36.11:19.45:44.44 was the best against each of the four fungi with significant interaction(P < 0.05) and good synergism being observed between the three oils. The results of this study demonstrated that mixture design could be useful to determine the optimum blend ratio of different essential oils for better antimicrobial effect at lower dosage, thereby providing methodological guidance for saving the dosage of natural bacteriostatic agents when used to preserve fruits and vegetables.
引文
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[2]SHELEF L A.Antimicrobial effect of spices[J].Journal of Food Safety,2007,6(1):29-44.DOI:10.1111/j.1745-4565.1984.tb00477.x.
[3]CEYLAN E,FUNG D Y C.Antimicrobial activity of spices[J].Journal of Rapid Methods&Automation in Microbiology,2004,12(1):1-55.DOI:10.1111/j.1745-4581.2004.tb00046.x.
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[5]INOUYE S,TAKIZAWA T,YAMAGUCHI H.Antibacterial activity of essential oils and their major constituents against respiratory tract pathogens by gaseous contact[J].Journal of Antimicrobial Chemotherapy,2001,47(5):565-573.DOI:10.1093/jac/47.5.565.
[6]CHUANG P H,LEE C W,CHOU J Y,et al.Anti-fungal activity of crude extracts and essential oil of Moringa oleifera Lam[J].Bioresource Technology,2007,98(1):232-236.DOI:10.1016/j.biortech.2005.11.003.
[7]GONCALVES M J,CRUZ M T,CAVALEIRO C,et al.Chemical,antifungal and cytotoxic evaluation of the essential oil of Thymus zygis subsp.sylvestris[J].Industrial Crops&Products,2010,32(1):70-75.DOI:10.1016/j.indcrop.2010.03.005.
[8]ZUZARTE M,GONCALVES M J,CAVALEIRO C,et al.Antifungal and anti-inflammatory potential of Lavandula stoechas,and Thymus herba-barona,essential oils[J].Industrial Crops&Products,2013,44(2):97-103.DOI:10.1016/j.indcrop.2012.11.002.
[9]BASSOLE I H,JULIANI H R.Essential oils in combination and their antimicrobial properties[J].Molecules,2012,17(4):3989-4006.DOI:10.3390/molecules17043989.
[10]STEINBERG J P.Superbug:the fatal menace of MRSA[J].Lancet Infectious Diseases,2010,10(8):519-520.DOI:10.3201/eid1610.101108.
[11]吕媛,李耘,郑波.国内外细菌耐药监测研究介绍[J].中国临床药理学杂志,2011,27(4):311-317.DOI:10.3969/j.issn.1001-6821.2011.04.021.
[12]葛永红,李灿婴,吕静祎,等.植物精油在果蔬采后病害控制中的研究进展[J].食品工业科技,2017,38(2):370-373.DOI:10.13386/j.issn1002-0306.2017.02.063.
[13]YAP P S,LIM S H,HU C P,et al.Combination of essential oils and antibiotics reduce antibiotic resistance in plasmid-conferred multidrug resistant bacteria[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,2013,20(8/9):710-713.DOI:10.1016/j.phymed.2013.02.013.
[14]YAP P S X,YIAP B C,PING H C,et al.Essential oils,a new horizon in combating bacterial antibiotic resistance[J].Open Microbiology Journal,2014,8(1):6-14.DOI:10.2174/1874285801408010006.
[15]TRIPATHI P,DUBEY N K,SHUKLA A K.Use of some essential oils as post-harvest botanical fungicides in the management of grey mould of grapes caused by Botrytis cinerea[J].World Journal of Microbiology and Biotechnology,2008,24(1):39-46.
[16]OUEDRHIRI W,BALOUIRI M,BOUHDID S,et al.Mixture design of Origanum compactum,Origanum majorana,and Thymus serpyllum,essential oils:optimization of their antibacterial effect[J].Industrial Crops&Products,2016,89:1-9.DOI:10.1016/j.indcrop.2016.04.049.
[17]HYLDGAARD M,MYGIND T,MERYER R L.Essential oils in food preservation:mode of action,synergies,and interactions with food matrix components[J].Frontiers in Microbiology,2012,3(12):12-24.DOI:10.3389/fmicb.2012.00012.
[18]HAMMER K A,CARSON C F.Antibacterial and antifungal activities of essential oils[M]//THORMAR H,LEY W,WILEY J.Lipids and Essential Oils as Antimicrobial Agents.US:A John Wiley and Sons,Ltd.,Publication,2011:255-306.DOI:10.1002/9780470976623.ch11.
[19]GUTIERREZ J,BARRY-RYAN C,BOURKE P.The antimicrobial efficacy of plant essential oil combinations and interactions with food ingredients[J].International Journal of Food Microbiology,2008,124(1):91-97.DOI:10.1016/j.ijfoodmicro.2008.02.028.
[20]TSERENNADMID R,TAKO M,GALGOCZY L,et al.Antibacterial effect of essential oils and interaction with food components[J].Central European Journal of Biology,2010,5(5):641-648.DOI:10.2478/s11535-010-0058-5.
[21]李伟,封丹,陆占国.孜然精油成分及其抗菌作用[J].食品科技,2008,33(5):182-186.DOI:10.13684/j.cnki.spkj.2008.05.077.
[22]CAKIR A,KORDALI S,ZENGIN H,et al.Composition and antifungal activity of essential oils isolated from Hypericum hyssopifolium and Hypericum heterophyllum[J].Flavour&Fragrance Journal,2004,19(1):62-68.DOI:10.1002/ffj.1279.
[23]HABA E,BOUHDID S,TORREGO-SOLLANA N,et al.Rhamnolipids as emulsifying agents for essential oil formulations:antimicrobial effect against Candida albicans and methicillin-resistant Staphylococcus aureus[J].International Journal of Pharmaceutics,2014,476(1/2):134-141.DOI:10.1016/j.ijpharm.2014.09.039.
[24]TIAN Jun,BAN Xiaoquan,ZENG Hong,et al.In vitro and in vivo activity of essential oil from dill(Anethum graveolens L.)against fungal spoilage of cherry tomatoes[J].Food Control,2011,22(12):1992-1999.DOI:10.1016/j.foodcont.2011.05.018.
[25]GOUPY J,CREIGHTON L.Introduction aux plans d’expériences[M].3rd ed,Paris:Dunod,2006:1-336.
[26]TENEVA D,DENKOVA Z,GOURANOV B,et al.Chemical composition and antimicrobial activity of essential oils from black pepper,cumin,coriander and cardamom against some pathogenic microorganisms[J].Acta Universitatis Cibiniensis,2016,20(2):39-52.DOI:10.1515/aucft-2016-0014.
[27]李大强.孜然精油的成分分析、抗菌和抗氧化活性[D].兰州:甘肃农业大学,2012:19-21.
[28]唐裕芳,唐小辉,张妙玲,等.花椒挥发油化学组成及抑菌活性研究[J].湘潭大学自然科学学报,2013,35(2):64-69.DOI:10.13715/j.cnki.nsjxu.2013.02.002.
[29]陈密玉,林燕妮,吴国欣,等.提取方法对芥子挥发油化学成分的影响[J].海峡药学,2005,17(5):71-74.DOI:10.3969/j.issn.1006-3765.2005.05.037.
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