复方五倍子有效成分的分离鉴定及抑菌活性研究

对渔用抗菌剂复方五倍子有效成分进行薄层色谱与柱层析分离检测,以鉴定出复方中的各种有效成分;选用几种水产动物常见致病菌进行体外抑菌和联合用药试验,以确定各有效成分抑菌活性强弱和各成分间的药物关系,为分析复方五倍子抗菌剂的药物作用机理奠定理论基础,并为其质量控制提供参考依据。用薄层析硅胶GF254制板,以苯:乙醇:甲酸(7:3:1)为展开剂,用氨熏-三氯化铁显色,日光下观察层析结果;通过干法装硅柱,以苯:乙醇:甲酸(7:3:1)作为洗脱液进行洗脱,分离得到各单体,参照理化性质分析及薄层分析,鉴定出复方五倍子抗菌剂总有效成分。结果表明;复方五倍子抗菌剂有效成分为鞣质、没食子酸、槲皮素、黄芩苷、芦荟大黄素、大黄素和1,8-二羟基蒽醌,其中没食子酸含量最大,紫外分光光度法测定为10.47%。用复方的各有效成分别对温和气单胞菌、豚鼠气单胞菌、迟缓爱德华菌和柱状黄杆菌进行抑菌试验,以温和气单胞菌为菌种将复方主要有效成分两两配对进行体外联合抑菌试验,得出复方五倍子各有效成分抑菌活性顺序为:没食子酸>蒽醌(含1,8-二羟基蒽醌、大黄素、芦荟大黄素)>黄芩苷>槲皮素>鞣质。没食子酸和蒽醌具有协同作用,没食子酸和黄芩苷、没食子酸和槲皮素、蒽醌和黄芩苷三组同属于累加作用。  

STUDIES ON SEPARATION AND ANTIBACTERIAL ACTIVITY OF THE EFFECTIVE COMPONENTS OF CHINESE HERBAL COMPOUNDS GALLNUT FOR FISHERY ANTIMICROBIAL AGENT

The effective components of Chinese Herbal Compounds Gallnut for fishery antimicrobial agent are separated and identified by thin-layer chromatography and column chromatography. Through antibacterial test and joint drug test of several aquatic animal pathogens in vitro, the antibacterial strength of active ingredients of the compound and the inter-relationship between the drug components are determined. The mechanism of Chinese Herbal Compounds Gallnut is supplied with the theoretical basis and the quality control is provided for its reference. The lamina is made with silica gel GF254 and the developing agent is spread with benzene: ethanol: formic acid (7:3:1). Moreover, it is colorized with NH3-FeCl3 and the result should be observed under sunlight. The monomer is isolated through dry silicon column loaded and it is eluted with benzene: ethanol: formic acid (7:3:1). The active ingredients of Chinese Herbal Compounds Gallnut are identified for physical and chemical properties and by thin-layer chromatography analysis. The results shows that the effective components of Chinese Herbal Compounds Gallnut are tannins, gallic acid, quercetin, baicalin, aloe emodin, emodin, 1, 8-2 hydroxy anthraquinone. The most content monomer is gallic acid by UV spectrophotometry for 10.47%. The antibacterial action for Aeromonas sobria, A. caviae, Edwrdsieda torda and flavobacterium columnare is tested. Aeromonas sobria selected as the object and the main active ingredients for two matched, antibacterial action in vitro is tested. Antibacterial order of the active ingredients is as follows: gallic acid > anthraquinone > baicalin > quercetin > tannide. There is synergy between gallic acid and anthraquinone, there is cumulative effect between gallic acid and baicalin, gallic acid and quercetin, anthraquinone and baicalin, respectively.