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亳州疏毛罗勒挥发油化学成分及抗氧化与抑菌活性研究



全 文 :  [收稿日期]2016-04-18
  [基金项目]安徽省质量工程—药学专业省级教学团队项目 (2013jxtd067);安徽省质量工程—药品质量检测技术特色专业项目
(2014tszy055);安徽省高等学校自然科学研究项目重点课题 (KJ2015A358);亳州市中药科技创新团队项目
(BZ2014zzb21)。
  [作者简介]刘超祥 (1972-),男,博士,副教授,主要从事天然产物化学成分及生物活性研究,cxliu888@163.com。
[引著格式]刘超祥,丁锐,王珊,等 .亳州疏毛罗勒挥发油化学成分及抗氧化与抑菌活性研究 [J].长江大学学报 (自科版),2016,
13 (21):34~37.
   亳州疏毛罗勒挥发油化学成分及抗氧化与抑菌活
性研究
  刘超祥,丁锐  (亳州职业技术学院药学院,安徽 亳州236800)
  王珊  (亳州职业技术学院药学院,安徽 亳州236800;安徽中医药大学药学院,安徽 合肥230000)
  丁连峰  (安徽济人药业有限责任公司,安徽 亳州236800)
  刘耀武  (亳州职业技术学院药学院,安徽 亳州236800)
[摘要]采用水蒸汽蒸馏法提取疏毛罗勒 (Ocimum basilicumL)挥发油,并采用气-质联用方法分析其化
学成分,再采用DPPH·清除能力测定法和O-2 ·清除能力测定法测定其抗氧化活性,杯碟法测定其抑菌
活性。结果表明,疏毛罗勒挥发油组分共检出27种成分,所鉴定出化学成分的质量占总量的90.10%,
其主要化学成分是十六酸乙酸乙酯 (44.16%)、乙酸松油酯 (17.1%)、(s)-2,5,5,8-四甲基-1,2,3,6-四
氢化薁-4-酮(9.92%)、香叶醇(3.05%)等;挥发油成分对DPPH·和O-2 ·有中等程度的清除能力;其对大
肠埃希菌、金黄色葡萄球菌、白色念珠菌3株菌种有一定的抑菌作用,并且浓度越高,抑菌效果越明显。
[关键词]疏毛罗勒 (Ocimum basilicumL);挥发油(;抗氧化性;抑菌活性
[中图分类号]Q949.777.6 [文献标识码]A   [文章编号]1673-1409 (2016)21-0034-04
疏毛罗勒 (Ocimum basilicumL)为唇形科罗勒属一年生全草植物,别名兰香、九层塔等。其味
辛甘、性温,有疏风解表、行气活血、解毒消肿之功效。主治感冒头痛、发热咳嗽、食积不化、中暑等
症[1,2],还可用于蛇虫咬伤和湿疹骚痒,且无毒副作用。近年来,罗勒的化学成分及生物活性研究引起
国内外科研工作者关注,疏毛罗勒药理学研究主要集中在降低血糖活性、抗应激活性、驱虫活性、治疗
腹泻、抗真菌活性和抗辐射引起的染色体损伤等领域[3,4]。
对亳州疏毛罗勒 (罗勒变种,俗称香花子)的实地调研和临床观察,发现亳州疏毛罗勒挥发油成分
对蚊虫叮咬引起的痛痒和炎症具有特殊疗效[5]。基于此,本研究对亳州地区疏毛罗勒挥发油的化学成分
分析、抗氧化及抑菌活性进行了研究,试图找到疏毛罗勒挥发油成分对蚊虫叮咬引起的痛痒与抑菌活
性、抗氧化活性之间的联系,为亳州地区疏毛罗勒的进一步开发利用提供参考依据。
1 材料与方法
1.1 材料
样品:采自安徽省亳州职业技术学院药用植物园,经安徽中医药大学方成武教授鉴定。
主要仪器:RE-52A旋转蒸发仪 (亚荣生化仪器厂产品),FA2004N电子天平 (菁海仪器有限公司
产品),752紫外可见分光光度计 (菁华科技仪器有限公司产品),SHZ- (Ⅲ)循环水式多用真空泵
(河南省予华仪器有限公司产品),HH-1数显恒温水浴锅 (晶波实验仪器厂产品),456-GC型气相色谱
·43·
长江大学学报 (自然科学版) 2016年7月 第13卷 第21期 (农学)
Journal of Yangtze University(Natural Science Edition) Jul.2016,Vol.13No.21
DOI:10.16772/j.cnki.1673-1409.2016.21.009
(Bruker公司生产),SCION单四级杆质谱仪 (Bruker公司生产),SPX-150B生化培养箱、MJX-160B
霉菌培养箱、HPX-9082MBE电热恒温培养箱 (均为上海博迅实业有限公司产品),AR5120电子分析
天平 (美国奥豪斯国际贸易有限公司产品)。
试剂:羟甲基氨基甲烷、DPPH·、抗坏血酸 (Vc)、邻苯三酚、0.9%无菌氯化钠溶液、丙酮等,
所用试剂均为分析纯。
菌株及培养基:大肠埃希菌 (Escherichia coli)、金黄色葡萄球菌 (Staphylococcus aureus)、枯草芽孢
杆菌 (Bacillus subtilis)、白色念珠菌 (Candida albicans)、营养肉汤培养基、营养琼脂培养基、改良马丁
(琼脂)培养基、氯化钠蛋白胨缓冲液 (pH 7.0)均由安徽济人药业有限责任公司检测中心提供。
1.2 方法
1.2.1 挥发油的提取
取疏毛罗勒200g,剪碎,装入500mL的圆底烧瓶中,水蒸气蒸馏提取挥发油[6],乙醚萃取,使用
旋转蒸发仪回收乙醚后,得到淡黄色透明油状物0.9g,挥发油得率0.45%,具有特殊香味,低温保存
备用。
1.2.2 化学成分分析
采用气-质联用技术对疏毛罗勒挥发油成分进行分析。
气相色谱条件:弹性毛细管色谱柱 (BR-17型,奥嵩科学仪器有限公司产品,0.25μm×250μm×
30m),载气:高纯氦气 (南京上元工业气体厂产品),纯度大于99.99%,柱流量1.0mL/min,采用分
流进样,分流比 10∶1,进样量1.0μL,进样口温度为 220℃;程序升温条件:初始温度为40℃
(2min),以2℃/min的速度升温至100℃ (保持2min),再以5℃/min的速度升温至250℃。
质谱条件:EI离子源,电子能量70eV,离子源温度为220℃,传输线温度为260℃,四级杆温度
为150℃;全扫描模式,扫描周期1s,质谱检索标准库:NIST 11.L。
1.2.3 抗氧化活性测定
1)O-2 ·自由基清除能力测定 参考文献 [7,8]的方法测试目标化合物清除超氧阴离子 (O-2 ·)
的能力。取50mmol/L的 Tris-HCl缓冲液 (pH 8.20)5.7mL,加入0.2mL浓度为0.1000g/L的样
品,置于25℃水浴保温10min,然后加入25℃预热6mmol/L邻苯三酚0.1mL,使总体积达到6.0mL,
迅速摇匀后用紫外可见分光光度计测定反应1min时320nm波长处光密度D,此后每隔30s读取1次光
密度D,300s时停止。以Tris-HCl缓冲液为空白调零,无样品液为阳性对照,测试所有样品,样品对
(O-2 ·)的抑制率按如下公式计算。用Vc进行空白对照[9]。
  抑制率 (I)= {[(△D/△t)对照- (△D/△t)]/(△D/△t)对照}×100%
2)清除DPPH·自由基能力测定 参考文献 [10,11]的方法测试目标化合物清除二苯代苦味酰
基自由基 (DPPH·)的能力。向1mL DPPH·乙醇溶液中加入不同浓度的疏毛罗勒挥发油使总体积
达到2mL。振荡器混匀后,室温,避光放置30min后,在518nm波长处测定光密度D1,DPPH·乙醇
溶液在518nm波长处测定光密度D0,平行测定3次,按下式计算清除率。用Vc进行空白对照。
  清除率= [(D0-D1)/D0]×100%
1.2.4 抑菌活性测定
采用杯碟法[12],检测疏毛罗勒挥发油对大肠埃希菌、金黄色葡萄球菌、枯草芽孢杆菌、白色念珠
菌的抑制作用。样品设5个浓度,分别为1050、525、262.5、131.25、65.625μg/mL。
2 结果与分析
2.1 疏毛罗勒挥发油化学成分
采用GC/MS分析对疏毛罗勒挥发油化学成分进行分析,共检测出27个峰。对总离子流图中的各
峰经质谱扫描后得到质谱图,经计算机质谱数据库检索按各色谱峰的质谱裂片图并结合有关文献对峰质
荷比和相对丰度等进行直观比较,同时还对一些主要成分采用标准物质对照,分别对各色谱峰加以确
·53·第13卷 第21期 刘超祥等:亳州疏毛罗勒挥发油化学成分及抗氧化与抑菌活性研究  
认,鉴定了疏毛罗勒挥发油组分中的化学成分,按面积归一法确定各组分相对含量,分析结果见表1。
表1 疏毛罗勒挥发油化学成分的GC/MS的分析结果
峰号 保留时间/s 化合物名称 分子式 质量分数/%
1  6.29 乙酸小茴香酯 C12H20O2 17.01
2  7.87 香叶醇 C10H18O  3.05
3  8.02 柠檬醛 C10H16O  0.28
4  8.23 乙酸龙脑酯 C12H20O2 0.51
5  8.42 顺-3-苯基丙烯酸甲酯 C10H10O2 2.35
6  8.49 香叶酸甲酯 C11H18O2 0.23
7  8.80 α-荜澄茄烯 C11H16 0.19
8  9.00 莰烯 C10H16O  1.32
9  9.09 古巴烯 C15H24 0.24
10  9.29 反-3-苯基-2-丙烯酸甲酯 C10H10O2 44.16
11  9.57 长叶烯 C15H24 0.53
12  9.63  1a,2,3,3a,4,5,6,7b-八氢-1,1,3a,7-四甲基-1-氢-环丙基[α]萘 C15H24 0.76
13  9.8 大根香叶烯B  C15H24 0.43
14  9.97  1,6-二甲基-4-(1-甲基乙基)-六氢萘 C15H24 0.28
15  10.06 (+)-双环倍半芹烯 C15H24 0.72
16  10.14 β-瑟林烯 C15H24 1.09
17  10.20  1,2,3,4,5,6,7,8,8a-八氢-1,4-二甲基-7-(1-异丙烯基)-甘菊环烃 C15H24 1.52
18  10.32  3-(1,3-丁间二烯基)-2,4,4-三甲基-环己烯 C13H18O  2.42
19  10.37 去氢白菖烯 C15H22 0.22
20  10.82  1,2,3,3a,4,5,8,8a-八氢-α.α.3,8-四甲基奥甲醇 C15H24 0.59
21  10.89 β-古芸烯 C15H24 0.61
22  11.02 二氢-8-开环古巴烯 C15H24 0.48
23  11.15 香树烯 C15H24 1.35
24  11.33 (s)-2,5,5,8-四甲基-1,2,3,6-四氢化薁 C14H20O  9.92
25  11.46 γ-杜松烯 C15H24 0.31
26  11.54  2-亚甲基-6,8,8-三甲基-三开环[5.2.2.0(1,6)]十一碳-3-醇 C15H24O  0.32
27  11.77  2,4,4,5-四甲基-3-(3-氧-1-丁烯基)-环己烯 C14H20O2 0.21
  由表1可知,疏毛罗勒挥发油成分鉴定出27种化合物,主要成分为萜类物质,占挥发油成分的
60.61%。相对含量较高的是十六酸乙酸乙酯 (44.16%)、酸松油酯 (17.01%)、(s)-2,5,5,8-四甲基-
1,2,3,6-四氢化薁-4-酮(9.92%)、香叶醇(3.05%)、3-(1,3-丁间二烯基)-2,4,4-三甲基-环己烯(2.42%)、
肉桂酸甲酯(2.35%)、香树烯(1.35%)、1,2,3,4,5,6,7,8,8a-八氢-1,4-二甲基-7-(1-异丙烯基)-甘菊环烃
(1.52%)、莰烯(1.32%)及B-瑟林烯(1.09%),其他成分含量较少。结合植物生源代谢途径分析,认为
疏毛罗勒挥发油成分主要通过甲戊二羟酸代谢合成途径、莽草酸代谢合成途径和乙酰-丙酮酸代谢合成
途径完成,没有发现氨基酸代谢合成途径产物。
2.2 疏毛罗勒挥发油的抗氧化活性
表2 疏毛罗勒挥发油抗氧化活性
成分 抑制率/% 清除率/%
挥发油 8.03  0.121×103
Vc  18.31  0.235×103
  由表2可知,疏毛罗勒挥发油具有一定的清除DPPH·自由
基的能力,但其清除能力没有其Vc的清除力好,即抗氧化作用
没有Vc强,且Vc的清除能力随浓度的稀释,其抗氧化作用也
逐渐降低。疏毛罗勒挥发油对于超氧阴离子O-2 ·具有一定的清
除能力,其清除能力随浓度的变化影响不大。
2.3疏毛罗勒挥发油的抑菌活性
从表3结果可以看出,疏毛罗勒挥发油样品有一定抑菌活性,对大肠埃希菌、金黄色葡萄球菌、枯
草芽孢杆菌、白色念珠菌均有一定的抑菌作用并成浓度依赖,浓度越高,抑菌作用越明显;但是对枯草
·63·   生物技术 2016年7月
芽孢杆菌的抑菌效果最差。
表3 疏毛罗勒挥发油抑菌活性的结果分析
菌种
样品浓度/(μg/mL)
1050  525  262.5  131.25  65.625
大肠埃希菌 76  70  58  42  23
金黄色葡萄球菌 50  43  32  23  11
枯草芽孢杆菌 60  54  48  46  43
白色念珠菌 54  44  34  28  22
3 结论
现代医学研究证实诸多疾病的发生和发展与自由基导致的机体细胞氧化损伤密切相关,从天然产物
中寻找安全有效的抗氧化剂成为近年来的研究热点之一。本研究以抗DPPH·自由基能力为指标,评价
疏毛罗勒挥发油成分的抗氧化能力;发现疏毛罗勒挥发油所含化学成分有较好的抗氧化性[13,14];其挥
发油对大肠埃希菌、金黄色葡萄球菌、白色念珠菌3株菌种随浓度升高抑菌效果越明显。试验过程中还
将其制成液体溶剂用于蚊虫叮咬实验,止痒效果也较明显。此外,课题组从疏毛罗勒挥发油组分中分离
纯化到一个具有驱蚊活性的单体化合物和一个具有止痒效果的单体化合物,对2个化合物的药理活性正
在进一步研究和开发。
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·73·第13卷 第21期 刘超祥等:亳州疏毛罗勒挥发油化学成分及抗氧化与抑菌活性研究  
was studied by market research and analysis.Combined with the questionnaire and SPSS statistical
methods,six dimensions of Shennongjia ecotourism experience quality were measured and analyzed,
which included ecotourism environment experience quality,service experience quality,information
service quality,facilities experience quality,tour price experience quality,tourist traffic experience
quality.The results showed that Shennongjia lacked of deep experience products;service quality could
not create a better emotional experience for tourists;tourism facilities could not create the atmos-
phere of ecotourism experience;tour price did not reflect the experience value of tourists;tourists ex-
pecte environmental education experience;tourists generaly relayed on web experience channels and
word of mouth publicity promotions.
Key words:tourism experience;Shennongjia;experience quality
025Comparative Test on Cultivation Methods of Large-sized Pelteobagrus fulvidraco Larvae
YI Jia-xin  Zhijiang Aquaculture Technology Extension Station,Zhijiang,Hubei 443200,China( )
YANG Jun,DONG Jian-feng ,FENG De-pin  Yichang Aquaculture Technology Extension Station,Yichang,Hubei 443000,China( )
Abstract:Large-sized Pelteobagrus fulvidraco larvae were cultivated with direct cultivation method
(control group)and graded cultivation method(experimental group)in six 0.3hm2 ponds under the
same condition.The stocking density of P.fulvidraco was three milion fries/hm2.According to the
results,after 60dof cultivation,the average survival rate of P.fulvidraco larvae in three ponds of
the control group reached 40.43%,with an average size of 8.18cm and an average yield of 9576.30
kg/hm2;the average survival rate of P.fulvidraco larvae in three ponds of the experimental group
reached 60.38%,with an average size of 9.44cm and an average yield of 18072.88kg/hm2.In conclu-
sion,the graded cultivation method was superior to the direct cultivation method.
Key words:Pelteobagrus fulvidraco;large-sized larvae;survival rate;yield;benefit
029Study on the Secondary Productivity of Macrozoobenthos in Binzhou Shalow Sea Waters
WANG Zhi-zhong,ZHANG Jin-lu
GONG Jun-xia  
Freshwater Fisheries Research Institute of Shandong Province,
Shandong Provincial Key Lab for Genetics and Breeding in Freshwater
Fisheries,Jinan,Shandong250013,China




SUN Tong-qiu,WANG Yu-qing
JING Zhong-hua,ZENG Hai-xiang 
Marine and Fisheries Research Institute of Binzhou City,
Binzhou,Shandong256616,China( )
Abstract:According to the results of macrozoobenthos in Binzhou shalow sea waters,with 3times
survey 18stance being made in the Spring,Summer and Autumn,2012,the secondary productivity
and P/Bvalue of macrozoobenthos were calculated by using Brey formula.The results showed that
the secondary productivity of macrozoobenthos in Binzhou shalow sea water was 0.002~65.00
g(AFDW)·m-2·a-1,the average was 6.21g(AFDW)·m-2·a-1,and its dry weight of biomass
was 0.001~43.68g(AFDW)·m-2,the average was 3.98g(AFDW)·m-2.Both of them were the
highest in Autumn,then in Spring,and it was the lowest in Summer.The P/Bvalue was higher,the
average was 1.80a-1,and it was the highest in Summer,then in Spring,and it was the lowest in Au-
tumn.The smal individual of short life history species had the advantage in the benthic animal com-
munity composition in this area,and the secondary productivity(wet weight)of large benthic animal
was about 69000tons every year.
Key words:Binzhou shalow sea water;macrozoobenthos;the secondary productivity;P/Bvalue
034Study on Antioxidant Activity and Antimicrobial Activity of Volatile Oils in the Ocimum
basilicumL
LIU Chao-Xiang,DING Rui  College of Pharmacy,Bozhou Vocational and Technical College,Bozhou,Anhui 236800 China( )
WANG Shan 
College of Pharmacy,Bozhou Vocational and Technical College,
Bozhou,Anhui 236800 China;College of Pharmacy,Anhui Traditional
Chinese Medicine University,Hefei,Anhui 230000,China




DING Lian-Feng (Anhui Jiren Pharmaceutical Industries Co.,Ltd,Bozhou,Anhui 236800,China)
LIU Yao-Wu (College of Pharmacy,Bozhou Vocational and Technical College,Bozhou,Anhui 236800 China)
Abstract:The constituents of volatile oil in the Ocimum basilicumL were analyzed by GC-MS meth-
od after it was extracted via steam distilation method.The antioxidant activity was assayed via scav-
enging properties of DPPH·and O-2 ·,the antibacterial activity was studied by using the cup-plate
method.The results showed that 27kinds of components were indeed recognized in volatile oils con-
stituent.The identified ingredients accounted for nearly 90.10% of weak polarity constituents,its

main chemical composition was hexadeanoic acid ethyl easter(44.16%),fenchyl acetate(17.1%),ge-
raniol(3.05%),(s)-2,5,5,8-tetramethyl-1,2,3,6-tetrahydro-azulen-4(5H)-one(9.92%),etc.The
volatile oil constituents had moderate activity for scavenging properties of DPPH·and O-2 · .The
constituents also had certain antibacterial activity against Escherichia coli,Staphylococcus aureus,
Bacillus subtilis and Candida albicans,and the higher the concentration was,the more obvious the
antibacterial activity was.
Key words:Ocimum basilicumL;volatile oil;antioxidant activity;antibacterial activity
042Study on Treatment of Swine Wastewater by Algae Technology
CHEN You-lan  College of Urban Construction,Yangtze University,Jingzhou,Hubei 434023,China( )
CAO Rong-hua  Chuandian Agricultural Service Center of Jingzhou City,Hubei Province,Jingzhou,Hubei 434033,China( )
Abstract:According to the characteristics of high concentration of nitrogen and phosphorus,organic
matter in swine wastewater,the algae biological treatment technology which meet the requirements
of technical specification was discussed.Through the design of orthogonal experiment,the effect of
filamentous algae on the treatment of swine wastewater was studied,and the optimum conditions
were obtained.According to the optimal conditions obtained by orthogonal experiment,the semi con-
tinuous treatment process was carried out to verify the effect of filamentous algae on the treatment of
swine wastewater.The results showed that the algae had a good treatment effect on the low carbon
and nitrogen ratio,high nitrogen and phosphorus content of swine wastewater.
Key words:swine wastewater;algae;nitrogen removal;phosphorus removal
047Study of the Status and Distribution of Crop Straw Resource in Hubei Province
LI Ji-fu,FU Guo-xing,MEI Qing-qing
LI Wen-jun,WANG Chang  
College of Agronomy,Yangtze University,Jingzhou,
Hubei 434025,China( )
Abstract:Based on statistical data and farm-household surveys,the annual crop straw types,distri-
bution and processing of crop straw in Hubei Province were analyzed.The results showed that the
main straw types were rice,rapeseed,wheat,corn and cotton straw,which accounted for 94%of the
total straw resource in the whole province.The research also indicated that crop straw was mainly
distributed in Jingzhou,Yichang,Xiangyang,Xiaogan,Jingmen and Huanggang,where the straw re-
sources were accounted for 72%of the total resource in 2014.Farm-household survey showed that
the ratio of straw returning to field was 32%,but there was significant difference among crops.The
proportion of rice and wheat straws were the highest by 54%and 43%,respectively.While the pro-
portion of burned and discarded remains high.Meanwhile,the current agricultural production was stil
smal-scale farming and widespread application of fertilizer-based.Therefore,direct retuning of straw
had great potential,which also was the most fundamental way to solve the straw utilization and in-
crease soil organic matter.
Key words:Hubei Province;crop straw resource;evaluation;straw returning to field;current situa-
tion
055Research on the Preservation of Crucian(Carassius auratus)by Extraction from Szechuan
Pepper(Zanthoxylum bungeanum Maxim)
SU Wei  College of Life Science,Yangtze University,Jingzhou,Hubei 434025,China( )
ZHOUJing  Department of Medical Services,The First Affiliated Hospital of Yangtze University,Jingzhou,Hubei 434000,China( )
CAORonghua  Chuandian Agricultural Service Center of Jingzhou City,Hubei Province,Jingzhou,Hubei 434033,China( )
WU Tao  College of Life Science,Yangtze University,Jingzhou,Hubei 434025,China( )
Abstract:Ultrasound assisted extraction of Szechuan pepper(Zanthoxylum bungeanum Maxim)
was used to the preservation of fresh crucian(Carassius auratus),pH,TVBNvalue,TBAvalue,to-
tal bacterial count and sensory score were used to study the effect of preservation quality.The results
showed that the extraction from szechuan pepper exhibited a beneficial effect on inhibiting the spoil-
age of fish filets,as a natural preservative,it had good potential for application.
Key words:crucian(Carassius auratus);szechuan pepper(Zanthoxylum bungeanum Maxim);pres-
ervation
Translated &Edited by SU Kai-ke(苏开科)