流动相对高效液相色谱法测氯霉素降解的影响  

The Effects of HPLC Mobile Phases on Chloramphenicol Degradation

作　　者：蒋瑞萍[1] 解开治[1] 孙丽丽[1] 卢钰升[1] 李夏[1] 顾文杰[1] 李文英[1] 

机构地区：[1]广东省农业科学院农业资源与环境研究所,农业部南方植物营养与肥料重点实验室,广东省养分资源循环利用与耕地保育重点实验室,广东广州510640

出　　处：《环境科学与技术》2016年第12期42-45,54,共5页Environmental Science & Technology

基　　金：广东省科技计划项目:广东省新型肥料与高效施肥工程技术研究中心建设(2014B090904068);广东省科技计划项目:广东省新型肥料科技创新与服务平台建设

摘　　要：为了更加高效、简便地评估摇瓶培养中降解菌对氯霉素（CAP）的降解效果,指导CAP高效降解菌株的筛选,建立以乙腈∶草酸（0.01 mol/L）=60∶40、甲醇∶水=60∶40流动相,用于摇瓶培养中无机盐培养基氯霉素含量的高效液相色谱法测定。结果表明,在0.5～100mg/L的浓度范围,采用乙腈-草酸作为流动相,其检出限为23μg/L,定量限为77μg/L,回收率为74.3%～93.8%,RSD为1.81%～14.69%;采用甲醇-水作为流动相,检出限为4.6μg/L,定量限为15μg/L,回收率为98%～103%,RSD为0.92%～3.25%。两者均适用于氯霉素降解菌的快速筛选,但甲醇-水作为流动相其灵敏度、准确度和精密性更高。Developing a simpler and more efficient method to evaluate the degradation capacity of strains to chloramphenicol（CAP）in shake-flask culture system is essential to screen CAP degrading strains. A high performance liquid chromatography（HPLC）method was established to determine the content of CAP in inorganic salts medium in shake-flask culture system.The effect of two different mobile phases,acetonitrile∶oxalic acid（0.01 mol/L）（60∶40）and methanol∶water（60∶40）,on the effective of CAP measurement was compared in the current study. Both the mobile phases had the same concentration range,from 0.5 to 100 mg/L. The detection limit,quantitation limit,recovery range and relative standard deviation range of CAP were 23 μg/L,77 μg/L,74.3%~93.8%,1.81%~14.69% respectively when acetonitrile-oxalic acid was used as HPLC mobile phase. In contrast,the detection limit,quantitation limit,recovery range and relative standard deviation range of CAP were 4.6 μg/L,15 μg/L,98% ~103%,0.92% ~3.25% respectively when methanol-water was used as HPLC mobile phase. Both HPLC mobile phases could be used for rapid screening of CAP-degrading bacteria. Furthermore,the methanol- water could provide higher sensitivity,more accurate and more precise test results compared with acetonitrile-oxalic acid,and thereby would be a comparatively ideal HPLC mobile phases for CAP-degrading bacteria screening.

关 键 词：高效液相色谱法 流动相 降解菌 摇瓶培养 

分 类 号：X132[环境科学与工程—环境科学]