汉麻叶提取物抑菌活性及抑菌稳定性研究
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摘要
目的研究汉麻叶提取物的抑菌活性及抑菌稳定性。方法用乙醇对汉麻叶成分提取并依次用石油醚、乙酸乙酯、正丁醇分级萃取,以4种常见食源性致病菌:金黄色葡萄球菌、大肠杆菌、单核细胞李斯特菌、沙门氏菌为指示菌,用牛津杯法和微量2倍稀释法测定汉麻叶提取物的抑菌活性并分别对其热稳定性、酸碱稳定性、紫外线稳定性、金属离子稳定性、蔗糖稳定性进行考察。结果汉麻叶提取物对大肠杆菌和沙门氏菌无抑制作用,但对金黄色葡萄球菌和单核细胞李斯特菌具有良好的抑菌作用,活性成分主要分布于乙酸乙酯相,其对2种细菌的最小抑菌浓度(minimum inhibitory concentration,MIC)分别为7.81mg/mL和15.63mg/mL;汉麻叶乙酸乙酯相经紫外线照射、蔗糖添加和不同温度处理仍表现出良好的稳定性,酸性条件使其对金黄色葡萄球菌和李斯特菌的抑菌活性增强,Ca~(2+)使其抑菌活性提高,Fe~(3+)使其抑菌活性降低,Na~+能够增强其对单核细胞李斯特菌的抑菌活性,K+使其对金黄色葡萄球菌的抑制作用有所增加。结论汉麻叶提取物具有良好的抑菌活性和抑菌稳定性,具备作为一种天然防腐剂进行开发和利用的前景。
Objective To study the antibacterial activity and stability of hemp(Cannabis sativa L.) leaves extract. Method The extraction of hemp leaves was extracted with ethanol and fractionated with petroleum ether, ethyl acetate and n-butanol. Four common foodborne pathogens: Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Salmonella were used as indicator bacteria. The antibacterial effect of extracts of hemp leaves was determined by Oxford cup method and trace 2-fold dilution method. The activity was investigated for its thermal stability, acid-base stability, UV stability, metal ion stability, and sucrose stability. Results Hemp leaf extract has no inhibitory effect on Escherichia coli and Salmonella, but it had good antibacterial activity against Staphylococcus aureus and Listeria monocytogenes. The active ingredient was mainly distributed in the ethyl acetate phase, and the minimum inhibitory concentration(MIC) for the 2 bacteria was 7.81 mg/mL and 15.63 mg/mL, respectively. The hemp leave extract by ethyl acetate phase showed good stability after UV irradiation, sucrose addition and different temperature treatments. The acidic conditions and Ca2+ enhanced the antibacterial activity against Staphylococcus aureus and Listeria, Fe3+ reduced its antibacterial activity, Na+ enhanced its antibacterial activity against Listeria monocytogenes, and K+ increased its inhibitory effect against Staphylococcus aureus. Conclusion Hemp leaf extract has good antibacterial activity and bacteriostatic stability, which has the prospect of being developed and utilized as a natural preservative.
Objective To study the antibacterial activity and stability of hemp(Cannabis sativa L.) leaves extract. Method The extraction of hemp leaves was extracted with ethanol and fractionated with petroleum ether, ethyl acetate and n-butanol. Four common foodborne pathogens: Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Salmonella were used as indicator bacteria. The antibacterial effect of extracts of hemp leaves was determined by Oxford cup method and trace 2-fold dilution method. The activity was investigated for its thermal stability, acid-base stability, UV stability, metal ion stability, and sucrose stability. Results Hemp leaf extract has no inhibitory effect on Escherichia coli and Salmonella, but it had good antibacterial activity against Staphylococcus aureus and Listeria monocytogenes. The active ingredient was mainly distributed in the ethyl acetate phase, and the minimum inhibitory concentration(MIC) for the 2 bacteria was 7.81 mg/mL and 15.63 mg/mL, respectively. The hemp leave extract by ethyl acetate phase showed good stability after UV irradiation, sucrose addition and different temperature treatments. The acidic conditions and Ca2+ enhanced the antibacterial activity against Staphylococcus aureus and Listeria, Fe3+ reduced its antibacterial activity, Na+ enhanced its antibacterial activity against Listeria monocytogenes, and K+ increased its inhibitory effect against Staphylococcus aureus. Conclusion Hemp leaf extract has good antibacterial activity and bacteriostatic stability, which has the prospect of being developed and utilized as a natural preservative.
引文
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[11]Diao WR,Zhang LL,Feng SS,et al.Chemical composition,antibacterial activity and mechanism of action of the essential oil from Amomum kravanh[J].J Food Prot,2014,77(10):1740-1746.
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[14]潘旭迟,许剑锋,傅昱晟,等.3种植物挥发油抑制食源性细菌生长活性成分及机理[J].食品科学,2017,38(13):143-149.Pan XC,Xu JF,Fu YS,et al.Antibacterial ingredients and mechanism of volatile oils from three kinds of plants against foodborne bacteria[J].Food Sci,2017,38(13):143-149.
[15]权美平,田呈瑞.水蒸气蒸馏法提取茜草精油抑菌活性及其机理的研究[J].食品与生物技术学报,2017,36(8):843-848.Quan MP,Tian CR.Study on the antibacterial activity and its mechanism of essential oil from madder(Rubia cordifolia L.)extracted by hdrodistillation[J].J Food Sci Biotechnol,2017,36(8):843-848.
[16]刘海燕,张建伟.明日叶提取物抑菌活性成分稳定性的研究[J].食品研究与开发,2016,37(10):39-42.Liu HY,Zhang JW.Study on stability of antibacterial activity compositions extracted from ashitaba[J].Food Res Dev,2016,37(10):39-42.
[17]赵磊,林文轩,迟茜,等.甜叶菊废渣提取物抑菌活性及抑菌稳定性研究[J].食品工业科技,2016,37(24):168-172.Zhao L,Lin WX,Chi Q,et al.Antibacterial activity and stability of stevia rebaudiana waste extract[J].Sci Technol Food Ind,2016,37(24):168-172.
[18]段伟丽,刘艳秋,包怡红.艾蒿精油的抑菌活性和稳定性[J].食品与生物技术学报,2015,34(12):1332-1337.Duan WL,Liu YQ,Bao YH.Study on antimicrobial activities and stability of essential oil from Artemisia argyi[J].J Food Sci Biotechnol,2015,34(12):1332-1337.
[19]张媛媛,冯亚净,李书国.五味子乙醇提取物的抑菌稳定性及抑菌机理研究[J].粮油食品科技,2016,24(1):55-60.Zhang YY,Feng YJ,Li SG.Study on the antimicrobial stability and antibacterial mechanism of ethanol extracts from fructus Schisandrae chinensis[J].Sci Technol Cereal Oil Food,2016,24(1):55-60.
[20]林树花,王伟,张菊华,等.10种植物精油对蓝莓病原菌的体外抑菌活性初筛[J].湖南农业科学,2018,(2):73-75.Lin SH,Wang W,Zhang JH,et al.The?initial?screening?of?ten?plant essential?oils?in?vitro?antibacterialactivity??of?blueberry?pathogens[J].Hunan Agric Sci,2018,(2):73-75.
[21]Oussalah M,Caillet S,Saucier L,et al.Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat[J].Meat Sci,2006,(73):236-224.
[22]Jallali I,Zaouali Y,Missaoui I,et al.Variability of antioxidant and antibacterial effects of essential oils and acetonic extracts of two edible halophytes:Crithmum maritimum L.and Inula crithmoides L.[J].Food Chem,2014,(145):1031-1038.
[23]王子怀,胡晓,李来好,等.罗非鱼水解蛋白金属离子螯合物的抑菌活性研究[J].食品工业科技,2014,35(8):79-82,87.Wang ZH,H X,Li LH,et al.Antibacterial activity of tilapia protein hydrolysates chelated with metal ions[J].Sci Technol Food Ind,2014,35(8):79-82,87.
[24]Mohammad J,Farideh TY,Seyed AM,et al.Quince seed mucilage films incorporated with oregano essential oil:Physical,thermal,barrier,antioxidant and antibacterial properties[J].Food Hydrocolloid,2014,(36):9-19.
[25]程晓强.东北山核桃壳黄酮的抑菌作用及产品开发[D].延吉:延边大学,2018.Cheng XQ.Antibacterial activity and products development of flavonoids from juglans mandshurica shell seed[D].Yanji:Yanbian University,2018.
[26]赵九阳,赵冠宇,刘玉梅.六氢β-酸/甲基-β-环糊精包合物的抑菌活性[J].精细化工,DOI:10.13550/j.jxhg.20180495Zhao JY,Zhao GY,Liu YM.Antimicrobial Activities of Hexahydro-β-Acids/Methyl-β-Cyclodextrin Inclusion Complex[J].Fine Chemicals,DOI:10.13550/j.jxhg.20180495
[27]胡唐玲,云云,徐志庆,等.白头翁汤正丁醇提取物抑制白念珠菌细胞膜[J].中国中药杂志,2017,42(16):3185-3190.Hu TL,Yun Y,Xu ZQ,et al.Butyl alcohol extract of Baitouweng decoction inhibits Candida albicans cell membrane[J].China J Chin Mater Medic,2017,42(16):3185-3190.
[28]刘艳芳,张德欣,丁寅寅.茄非食部分生物碱提取液抑菌活性研究[J].食品与机械,2018,34(1):67-71.Liu YF,Zhang DX,Ding YY.Study on bacteriostatic activities of alkaloid extract from the inedible parts of the eggplant[J].Food Mach,2018,34(1):67-71.
[2]Voukeng IK,Beng VP,Victor K.Antibacterial activity of six medicinal Cameroonian plants against Gram-positive and Gram-negative multidrug resistant phenotypes[J].BMC Complem Altern M,2016,16(1):388.
[3]张建春.汉麻综合利用技术[M].北京:长城出版社,2006.Zhang JC.Comprehensive utilization technology of hemp[M].Beijing:Great wall Publishers,2006.
[4]张树权,王贵江,宋宪友,等.黑龙江省汉麻产业发展的优势和对策[J].黑龙江农业科学,2018,(1):125-128Zhang SQ,Wang GJ,Song XY,et al.Advantages and countermeasures of the development of hemp industry in Heilongjiang province[J].Heilongjiang Agric Sci,2018,(1):125-128.
[5]Elsohly MA,Radwan MM,Gul W,et al.Phytochemistry of Cannabis sativa L[J].Prog Chem Org Nat Prod,2017,(103):1-36.
[6]杜军强,何锦风,何聪芬,等.汉麻叶活性成分及其药理特性的研究概况[J].中国医药导报,2011,8(31):9-11.Du JQ,He JF,He CF,et al.Review on active ingredients from Cannabis sativa leaves and its pharmacological characteristics[J].Chin Med Herald,2011,8(31):9-11.
[7]隽惠玲,史高峰,李春雷,等.低毒大麻叶茎浸膏抑菌作用的研究[J].安徽农业科学,2008,36(32):14168-14169,14196.Juan HL,Shi GF,Li CL,et al.Study on the bacteriostasis of the extract of low toxic Cannabis leaves and stems[J].J Anhui Agric Sci,2008,36(32):14168-14169,14196.
[8]崔广东,陈学福,史高峰,等.不同生长期的低毒大麻叶浸膏抑菌作用的研究[J].食品科技,2008,(2):192-194.Cui GD,Chen XF,Shi GF,et al.Study on the bacteriostatic action to the extract of different grow period low poison cannabis leaves[J].Food Sci Technol,2008,(2):192-194.
[9]Chakraborty S,Afaq N,Singh N,et al.Antimicrobial activity of Cannabis sativa,Thuja orientalis and Psidium guajava leaf extracts against methicillin-resistant staphylococcus aureus[J].J Integr Med,2018,16(5):350-357.
[10]Zou L,Hu YY,Chen WX.Antibacterial mechanism and activities of black pepper chloroform extract[J].J Food Sci Technol,2015,52(12):8196-8203.
[11]Diao WR,Zhang LL,Feng SS,et al.Chemical composition,antibacterial activity and mechanism of action of the essential oil from Amomum kravanh[J].J Food Prot,2014,77(10):1740-1746.
[12]贺怡琼,于宏伟,史宝胜,等.野生酸浆根茎叶提取物的抑菌活性及其稳定性研究[J].食品科技,2016,41(4):208-212.He YQ,Yu HW,Shi BS,et al.Antibacterial activity and stability of rhizome and leaf extract from wild Physalis[J].Food Sci Technol,2016,41(4):208-212.
[13]常曼曼,于宏伟,郭润芳,等.紫花地丁醇取物对食源性致病微生物的抑制作用[J].食品科技,2017,42(6):215-219.Chang MM,Yu HW,Guo RF,et al.Inhibition of extracts from Viola yedoensis Makino on food-borne pathogens[J].Food Sci Technol,2017,42(6):215-219.
[14]潘旭迟,许剑锋,傅昱晟,等.3种植物挥发油抑制食源性细菌生长活性成分及机理[J].食品科学,2017,38(13):143-149.Pan XC,Xu JF,Fu YS,et al.Antibacterial ingredients and mechanism of volatile oils from three kinds of plants against foodborne bacteria[J].Food Sci,2017,38(13):143-149.
[15]权美平,田呈瑞.水蒸气蒸馏法提取茜草精油抑菌活性及其机理的研究[J].食品与生物技术学报,2017,36(8):843-848.Quan MP,Tian CR.Study on the antibacterial activity and its mechanism of essential oil from madder(Rubia cordifolia L.)extracted by hdrodistillation[J].J Food Sci Biotechnol,2017,36(8):843-848.
[16]刘海燕,张建伟.明日叶提取物抑菌活性成分稳定性的研究[J].食品研究与开发,2016,37(10):39-42.Liu HY,Zhang JW.Study on stability of antibacterial activity compositions extracted from ashitaba[J].Food Res Dev,2016,37(10):39-42.
[17]赵磊,林文轩,迟茜,等.甜叶菊废渣提取物抑菌活性及抑菌稳定性研究[J].食品工业科技,2016,37(24):168-172.Zhao L,Lin WX,Chi Q,et al.Antibacterial activity and stability of stevia rebaudiana waste extract[J].Sci Technol Food Ind,2016,37(24):168-172.
[18]段伟丽,刘艳秋,包怡红.艾蒿精油的抑菌活性和稳定性[J].食品与生物技术学报,2015,34(12):1332-1337.Duan WL,Liu YQ,Bao YH.Study on antimicrobial activities and stability of essential oil from Artemisia argyi[J].J Food Sci Biotechnol,2015,34(12):1332-1337.
[19]张媛媛,冯亚净,李书国.五味子乙醇提取物的抑菌稳定性及抑菌机理研究[J].粮油食品科技,2016,24(1):55-60.Zhang YY,Feng YJ,Li SG.Study on the antimicrobial stability and antibacterial mechanism of ethanol extracts from fructus Schisandrae chinensis[J].Sci Technol Cereal Oil Food,2016,24(1):55-60.
[20]林树花,王伟,张菊华,等.10种植物精油对蓝莓病原菌的体外抑菌活性初筛[J].湖南农业科学,2018,(2):73-75.Lin SH,Wang W,Zhang JH,et al.The?initial?screening?of?ten?plant essential?oils?in?vitro?antibacterialactivity??of?blueberry?pathogens[J].Hunan Agric Sci,2018,(2):73-75.
[21]Oussalah M,Caillet S,Saucier L,et al.Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat[J].Meat Sci,2006,(73):236-224.
[22]Jallali I,Zaouali Y,Missaoui I,et al.Variability of antioxidant and antibacterial effects of essential oils and acetonic extracts of two edible halophytes:Crithmum maritimum L.and Inula crithmoides L.[J].Food Chem,2014,(145):1031-1038.
[23]王子怀,胡晓,李来好,等.罗非鱼水解蛋白金属离子螯合物的抑菌活性研究[J].食品工业科技,2014,35(8):79-82,87.Wang ZH,H X,Li LH,et al.Antibacterial activity of tilapia protein hydrolysates chelated with metal ions[J].Sci Technol Food Ind,2014,35(8):79-82,87.
[24]Mohammad J,Farideh TY,Seyed AM,et al.Quince seed mucilage films incorporated with oregano essential oil:Physical,thermal,barrier,antioxidant and antibacterial properties[J].Food Hydrocolloid,2014,(36):9-19.
[25]程晓强.东北山核桃壳黄酮的抑菌作用及产品开发[D].延吉:延边大学,2018.Cheng XQ.Antibacterial activity and products development of flavonoids from juglans mandshurica shell seed[D].Yanji:Yanbian University,2018.
[26]赵九阳,赵冠宇,刘玉梅.六氢β-酸/甲基-β-环糊精包合物的抑菌活性[J].精细化工,DOI:10.13550/j.jxhg.20180495Zhao JY,Zhao GY,Liu YM.Antimicrobial Activities of Hexahydro-β-Acids/Methyl-β-Cyclodextrin Inclusion Complex[J].Fine Chemicals,DOI:10.13550/j.jxhg.20180495
[27]胡唐玲,云云,徐志庆,等.白头翁汤正丁醇提取物抑制白念珠菌细胞膜[J].中国中药杂志,2017,42(16):3185-3190.Hu TL,Yun Y,Xu ZQ,et al.Butyl alcohol extract of Baitouweng decoction inhibits Candida albicans cell membrane[J].China J Chin Mater Medic,2017,42(16):3185-3190.
[28]刘艳芳,张德欣,丁寅寅.茄非食部分生物碱提取液抑菌活性研究[J].食品与机械,2018,34(1):67-71.Liu YF,Zhang DX,Ding YY.Study on bacteriostatic activities of alkaloid extract from the inedible parts of the eggplant[J].Food Mach,2018,34(1):67-71.