Study On The Interaction Between Benzimidazole Anthelmintics/fluoroquinolones And Probe By High Performance Liquid Chromatography Associating With Resonance Rayleigh Scattering And Its Application

The substances of various molecular weight,dissociation and non dissociation,volatile and non-volatile were separated by high performance liquid chromatography(HPLC),therefore,HPLC has been widely applied in various fields.For instance,biological chemistry,food analysis,pharmaceutical research,environmental analysis,inorganic analysis and so on.HPLC consists of reservoir,pump,sample injector,chromatographic column,detector and recorder,chromatographic column and detector play an important role.According to chromatographic separation mode,chromatographic column have reversed phase column,normal phase chromatographic columns,ion-change chromatography,hydrophobic interaction chromatography,size exclusion chromatography,affinity chromatography,chiral chromatography and so on.Reversed phase column was applied in this experiment.Ultraviolet detector,differential refractive index detector and fluorescence detector are most used in high performance liquid chromatography(HPLC).Fluorescence detector was applied in this experiment.In recent years,resonance Rayleigh scattering attracts more attention because it has a lot of advantages,for example,high sensitivity,easy to operate,higher determination speed and so on,but the selectivity is low.On the contrary,the selectivityof HPLC is good,but the sensitivity is lower than resonance Rayleigh scattering.Therefore,high performance liquid chromatography associating with resonance Rayleigh scattering has high selectivity and high sensitivity at the same time.The main study contents of this paper were as follows:1.Study on the interaction between three benzimidazole anthelmintics and eosin Y by high performance liquid chromatography associating with resonance Rayleigh scattering and its applicationA novel,highly selective,and sensitive resonance Rayleigh scattering(RRS)detection approach coupled with high performance liquid chromatography(HPLC)was researched and developed for the synchronous analysis of three kinds of benzimidazole anthelmintics,including mebendazole(MBZ),albendazole(ABZ),and fenbendazole(FBZ)for the first time.In the pH range of 3.5–3.7 Britton-Robinson buffer medium,three kinds of anthelmintics,which were separated by HPLC,reacted with eosin Y(EY)to form 1:1 ion-association complexes,resulting in significantly enhanced RRS signals and the maximum peak located at 335 nm.The enhanced RRS intensity was in proportion to the MBZ,ABZ,and FBZ concentration in the range0.2–25,0.2–23,and 0.15–20?g·m L-1,respectively.The limit of detection was in the range of 0.064–0.16?g·mL-1.In addition,human urine was determined to validate the proposed method by spiked samples and real urine samples.Satisfactory results were obtained by HPLC-RRS method.2.Simultaneous determination of three fluoroquinolones by the selective high performance liquid chromatography associating with sensitive resonance Rayleigh scattering and mechanism studyA novel and selective high performance liquid chromatography(HPLC)technique incorporating resonance Rayleigh scattering(RRS)detection approach was developed for the determination of three fluoroquinolones(FQS)synchronously for the first time,including enoxacin(ENO),ofloxacin(OFLX)and lomefloxacin(LMLX).In pH 4.4Britton-Robinson(BR)buffer medium,the ENO,OFLX,LMLX separated by HPLC reacted with erythrosine(ERY)to form 1:1 ion-association complexes,which led to significant enhancement of RRS.The maximum RRS wavelength of the ion-association complex located at 330 nm.The detection limits of ENO,OFLX,LMLX were5.1ng·mL-1,3.1ng·mL-1,4.2ng·mL-1 respectively at a signal-to-noise ratio of 3.In this work,the optimum experimental conditions and reaction mechanism were investigated in detail.The proposed method could be applied to the determination of ENO,OFLX,LMLX in water samples synchronously and the results were satisfactory.3.High performance liquid chromatography combined with resonance Rayleigh scattering for the detection of sparfloxacin and gatifloxacin in water samplesThe gatifloxacin(GFLX)and sparfloxacin(SPLX)separated by HPLC mixed with Britton-Robinson(BR)buffer medium of pH 4.4,then,GFLX and SPLX reacted with erythrosine(ERY)to form 1:1 ion-association complexes,which led to significant enhancement of RRS.Based on this principle,a novel analytical method of HPLC-RRS was developed for the determination of GFLX and SPLX.The flow rate of HPLC was 0.4mL·min-1,the temperature of reversed phase column was 40? and injection volume of GFLX and SPLX were 20?L.On the other hand,the optimum experimental conditions were investigated in detail,including reaction temperature,reaction tube length,flow rate of BR and ERY,detection wavelength and so on.The reasons for RRS enhancement were studied.The maximum RRS wavelength of the ion-association complex located at 330 nm,the detection limits of GFLX and SPLX were 3.8ng·mL-1and 17.5ng·mL-1 respectively at a signal-to-noise ratio of 3.The proposed method could be applied to the determination of GFLX and SPLX in water samples synchronously and the results were satisfactory.