基于固相萃取及高效液相色谱-荧光检测分析的污泥中氟喹诺酮类抗生素研究方法的开发

Development of Determination Method of Fluoroquinolone Antibiotics in Sludge Based on Solid Phase Extraction and HPLC-Fluorescence Detection Analysis

氟喹诺酮类抗生素（FQs）作为常见的药品和个人护理用品（PPCPs）在环境中广泛存在.由于在生物体内很难被代谢,FQs会通过粪尿排泄等途径进入城市污水处理系统并最终富集到污泥中.并且,由于其在环境中的累积效应,FQs的存在极易增强致病菌的耐药性,限制了污泥的后续资源化利用.为得到适用于污泥中FQs的分析方法,进一步把握FQs在污水及污泥处理过程中的固/液分配及降解规律,研究中选择4种代表性FQs：氧氟沙星（OFL）、诺氟沙星（NOR）、环丙沙星（CIP）和洛美沙星（LOM）为研究对象,并通过所设计的正交试验,考察了固相萃取材料、萃取体系p H、洗脱剂对FQs的固相萃取效率的影响,在此基础上进一步优化了固相样品中FQs的提取方法,最终建立了细胞破碎,碱式提取（三乙胺/甲醇/水以5/25/75混合）,固相萃取富集（SPE）,磷酸-三乙胺溶液为缓冲盐流动相,甲醇为有机流动相,梯度荧光扫描的高效液相色谱-荧光（HPLCFLD）检测方法.液态样品中4种FQs的加标回收率采用新方法后可达到82%-103%;固态样品中4种FQs的加标回收率也可达到71%-101%.同时,为进一步确认不同运行条件对污泥吸附FQs能力的影响,针对好氧、缺氧和厌氧污泥分别进行了吸附试验研究,发现处于不同活性状态的污泥（厌氧、缺氧和好氧污泥）对FQs的吸附能力呈现出递减的趋势,但对FQs的吸附率都达到90%以上.该结论也证实了实际污水处理厂所去除的50%的FQs主要是通过污泥吸附这一途径实现的.

Fluoroquinolone antibiotics（ FQs）,as the common pharmaceuticals and personal care products（ PPCPs）,are widespread in the environment. FQs contained in wastewater would be ultimately enriched in sludge,posing a potential threat to the consequent sludge utilization. To optimize the analytical method applicable to the determination of FQs in sludge,the authors selected ofloxacin（ OFL）,norfloxacin（ NOR）,ciprofloxacin（ CIP） and lomefloxacin（ LOM） as the target FQs,and established a method which was based on cell lysis,FQs extraction with triethylamine / methanol / water solution,Solid Phase Extraction（ SPE） and HPLC-Fluorescence Detection（ FLD） determination. After the investigation,phosphoric acid-triethylamine was decided to be the buffer salt,and methanol was chosen as the organic mobile phase. The gradient fluorescence scanning strategy was proved to be necessary for the optimal detection as well. Furthermore,by the designed orthogonal experiments,the effects of the extraction materials,p H,and the eluents on the efficiency of SPE extraction were evaluated,by which the optimal extraction conditions were determined. As a result,FQs in liquid samples could be analyzed by utilizing HLB extraction cartridge,and the recovery rates of the four FQs were in the range of 82%-103%. As for solid samples,the recovery rates of the four FQs contained reached up to 71%-101%. Finally,the adsorptivity of the sludge from the different tanks（ anaerobic,anoxic and oxic tanks） was investigated,showing gradual decrease in the adsorption capacity,but all adsorbed over 90% of the FQs. This conclusion also confirmed that 50% removal of FQs in the domestic wastewater treatment plant was realized by sludge adsorption.