摘要
为了分离姜厚朴中的单体化合物和厚朴酚,并探究其抑菌效果,试验采用高效液相色谱筛选色谱条件,以保留时间、色谱图及标准曲线进行分离物的定性和定量分析;对提取得到的和厚朴酚运用K-B纸片法对9种微生物进行抑菌圈直径的测定,将筛选得到的对和厚朴酚敏感的菌株用微量肉汤稀释法测定最小抑菌浓度(MIC)、最小杀菌浓度(MBC)。结果表明:以60%乙醇为最佳提取溶剂,以乙酸乙酯-石油醚(5∶5,V∶V)为最佳萃取条件,甲醇-0.5%甲酸水(80∶20,V∶V)等度洗脱为最佳色谱收集条件。结合保留时间和色谱图,确定收集物为和厚朴酚单体化合物,绘制的标准曲线方程为Y=1.15×10^(7)X+5.91×10^(4),R^(2)=0.998,线性范围为0.0016~1 mg/mL,计算得到收集的和厚朴酚单体化合物纯度为85.1%。药效学评价和厚朴酚对大肠杆菌、溶血性大肠杆菌、沙门氏菌、猪霍乱沙门氏菌、鼠伤寒沙门氏菌、肺炎克雷伯菌无抑制作用,对粪肠球菌、无乳链球菌、金黄色葡萄球菌有抑制作用,抑制圈直径分别为(15.00±0.16)mm、(17.00±0.20)mm、(19.00±0.23)mm,MIC、MBC分别为375.00,93.75,187.50μg/mL和750.00,375.00,375.00μg/mL。说明本试验提取得到的单体化合物和厚朴酚对金黄色葡萄球菌、粪肠球菌、无乳链球菌有抑制作用。
In order to isolate the monomer compound honokiol from ginger mix-fried magnolia bark, and to explore its antibacterial effect, in the experiment, high performance liquid chromatography was used to screen the chromatographic conditions, and the qualitative and quantitative analysis of the isolates was carried out based on retention time, chromatogram and standard curve. For the extracted honokiol, K-B disc method was used to measure the diameters of inhibition zones against nine microorganisms;MIC and MBC of the strains sensitive to honokiol obtained by screening were determined by micro-broth dilution method. The results showed that 60% ethanol was the best extraction solvent;ethyl acetate-petroleum ether(5∶5, V∶V) was the best extraction condition, and methanol with 0.5% formic acid water(80∶20, V∶V) for isocratic elution was the best chromatographic collection condition. Combined with the retention time and chromatogram, the collected material was confirmed to be the monomer compound honokiol. The drawn standard curve equation was Y=1.15×10^(7)X+5.91×10^(4),R^(2)=0.998;the linear range was 0.001 6-1 mg/mL, and the purity of the collected monomer compound honokiol was calculated to be 85.1%. Pharmacodynamic evaluation showed that it had no inhibitory effect on Escherichia coli, hemolytic Escherichia coli, Salmonella, Salmonella enterica serovar Choleraesuis, Salmonella enterica serovar Typhimurium, Klebsiella pneumoniae, but had inhibitory effect on Enterococcus faecalis, Streptococcus agalactiae, Staphylococcus aureus, with inhibition circle diameters of(15.00±0.16)mm,(17.00±0.20)mm and(19.00±0.23)mm, respectively, and with MIC and MBC values of 375.00, 93.75, 187.50 μg/mL, and 750.00, 375.00, 375.00 μg/mL, respectively. The results suggested that the monomer compound honokiol extracted in this experiment had inhibitory effect on Staphylococcus aureus, Enterococcus faecalis and Streptococcus agalactiae.
作者
邬佳莉
朱茂银
任胜杰
侯明鹏
申宏利
常华
项勋
段纲
WU Jiali;ZHU Maoyin;REN Shengjie;HOU Mingpeng;SHEN Hongli;CHANG Hua;XIANG Xun;DUAN Gang(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2022年第20期115-120,128,共7页
Heilongjiang Animal Science And veterinary Medicine
基金
云南省石林县奶山羊产业科技特派团项目(202104BI090017)
云南农业大学兽医公共卫生省创新团队项目(202105AE160014)
云南省农业基础研究联合专项面上项目(2018FG001-043)
云南省基础研究面上项目(202001AT070119)。
关键词
姜厚朴
高效液相色谱
和厚朴酚
抑菌试验
萃取条件
定性定量分析
ginger mix-fried magnolia bark
high performance liquid chromatography
honokiol
antibacterial test
extraction condition
qualitative and quantitative analysis
