| High-performance liquid chromatography?HPLC?is one of the most widely used branches of chromatography.In the device of HPLC,the detector has very important function,frequently used detectors include electrochemical detectors,ultraviolet detectors,and fluorescence detectors,etc.resonance Rayleigh scattering?RRS?was first reported as a novel and highly sensitive analytical method.Because of the method is simple and analysis speed fast high sensitivity is the most important,which has been widely applied to many fields such as pharmaceutical analysis,inorganic,biological macro-molecules,etc.RRS has a poor selectivity,which is easily interfered by similar matrix and cannot accurately detect various substances,however,HPLC has high selective advantage,but its detection sensitivity is not high,Therefore,high performance liquid chromatography and resonant Rayleigh scattering?HPLC-RRS?are used to combine the advantages and advantages of the two methods to obtain a highly selective and highly sensitive method.It also promotes the further development of the two methods,the main contents of this thesis are as follows:1.A technology of high performance liquid chromatography associated with resonance Rayleigh scattering was applied to determine three trichomoniasis drugs in honey and mechanism study.In this paper,a high sensitivity and high selectivity method is established:high performance liquid chromatography combined with resonance Rayleigh scattering?HPLC-RRS?,and used to detect three kinds of trichomoniasis drugs in honey,they are Metronidazole?MNZ?,ornidazole?ONZ?and inidazole?TNZ?.In pH 3.8 BR buffer solution,the three anti-trichomonas drugs separated by HPLC,separately reacted with a post-column derivatization Phloxine B?PB?in the reaction to form1:1 ion associationin,resulting in significantly enhanced resonance Rayleigh scattering signal.The volume ratio of the mobile phase is acetonitrile:potassium dihydrogen phosphat ewith a pH of 3.0=30:70,and the flow rate was 0.5ml.min-1.column temperature was35?,detection wavelength was?ex=?em=375 nm.This paper optimized the experimental conditions of post-column derivation and the mechanism of reaction for the enhancement of resonance Rayleigh scattering signal are discussed.Under the optimal experimental conditions,the RRS signals of the three anti-trichomonas drugs were enhanceed with the increase of drug concentration in the respective linear range.The detection limit?S/N?of metronidazole,ornidazole and tinidazole respectively was:0.013?g·mL-1,0.049?g·mL-11 and 0.27?g·mL-1,the recovery rate was 97.4%102.2%,and the relative standard deviation?RSD?does not exceed 5.3%.So the result of this method is satisfactory,and the reliability of this technique is verified.2.Study on the technology high performance liquid chromatography associated with resonance Rayleigh scattering basing on the Pd?II?-erythrosine as probe for determination of H2-receptor antagonist in human urine.In this paper,the technique of high performance liquid chromatography and resonance Rayleigh scattering?HPLC-RRS?was used to establish a new post-column derivation method,the metal ion Pd?II?and Ery dye were used as probes to introduce the reaction tube.This technique has greatly improved the sensitivity of the method and has been used for the first time to detect the two H2-receptor antagonist including famotidine?FMTD?and RNTD.In pH=4.6 BR buffer solution,FMTD and RNTD can be combined with Pd?II?to form cationic ions with a 1:1 ratio,and then reacted with the Ery by electrostatic attraction and hydrophobic to beionic compound[Pd?FMTD?]?Ery?2.Separation conditions of HPLC including the volume ratio of acetonitrile and sodium acetic acid?pH=4.5?was 10:90,flow rate of 0.5 mL·min-1,C18 column,and column temperature was 35?.This paper optimized the conditions of the experiment,such as the influence of organic solvent,detection wavelength,reaction tube length,reaction temperature,concentration of Pd?II?and Ery,etc.This method was used to obtain the detection limit?S/N?of FMTD and RNTD?S/N?were:7.3 ng·mL-1,10.1 ng·mL-1,the recovery rate was between 97.76%and 98.82%.Finally,In this experiment conditions,the method was successfully applied to detect the contents of FMTD and RNTD in human urine,there was no other matrix in the urine to interfere the RRS signal.Therefore,the method is reliable and practical.3.Study on the interaction of the nootropic-Palladium?II?combined with the congo red by fluorescence and resonance Rayleigh scattering and their analytical application.A highly sensitive detection approach of resonance Rayleigh scattering spectra?RRS?is firstly applied to analyzing nootropic drugs including piracetam?PIR?and oxiracetam?OXI?.In pH=3.0 HCl-NaAc buffer solution,OXI chelated with palladium?II?to form the cation[Pd2·OXI]2+,and then reacted with Congo red?CGR?by virtue of electrostatic attraction and hydrophobic force to form binary complex[Pd2·OXI].CGR2,which could result in the great enhancement of RRS.Resonance Rayleigh scattering signal of the system is recorded at?ex=?em=375nm.This mixture complex not only has higher RRS,but also makes contribution to significant increase of fluorescence,and the same phenomena also were discovered in PIR.The enhanced RRS intensity was proportional to the concentration of nootropic in the range of 0.03?g·mL-13.0?g·mL-1,the detection limit?DL?of PIR and OXI detected by RRS method is is 2.3ng·mL-11 and 9.7 ng·mL-1.In addition,the DL of fluorescence method to detect PIR and OXI is 8.4?g·mL-11 and 19.5?g·mL-1.Obviously,the RRS is the highly sensitive method,The recoveries of nootropic drugs were 100.4%101.8.0%and the RSD?n=5?was 1.1%3.1%by RRS method.This paper not only investigated the optimum conditions for detecting nootropics with using RRS method,but also focused on exploring the mechanism of RRS signal and the reasons for its enhancement.Finally,the RRS method has successfully detected nootropic drugs in human urine samples with satisfactory results. |
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