| Objective: Ofloxacin,belongs to quinolones,has broad-spectrum antibacterial characteristics.It has a strong antibacterial activity for the great majority of bacteria and extensively clinical application.Ofloxacin is one of chiral drugs.At present,the reports on its assessment are all based on the study of racemic carries,and there have not been the reports to reasearch two enantiomers caused by the drug into the body,which results in lower or higher evaluation and brings the hidden dangers to the rational use of drugs and human health.Therefore,The study for drug has beenthe research hotspot in the field of chemistry and medicine.The purpose of this study is to establish the method to detect racemic ofloxacin and two enantiomers in high performance liquid chromatography and the method to test the content of ofloxacin by charge transfer reaction with microdialysis sequential injection,which provide a theoretical basis for the rational use of ofloxacin in clinic and technical support for the realization of online detection of ofloxacin.Methods: The content of ofloxacin blood of rats was tested by microdialysis sampling technique combined with high performance liquid chromatography.The content and distribution of two kinds of enantiomers in mouse blood and tissues,after administration of ofloxacin was dectected with copper sulfate solution and Dphenylalanine as chiral mobile phase additive to establish the high performance liquid chromatography.The content of ofloxacin in rat blood content was tested based on the charge transfer reaction of ofloxacin and chloranilic acid combination of microdialysis and μSIA-LOV.Results:(1)The reversed-phase high performance liquid chromatography(HPLC)method for the determination of the content of ofloxacin in blood of rats was established.From 0.025μg/m L to 3.200μg/mL,the content of ofloxacin and peak area showed good linear relationship(r=0.9996).The RSD values of inter day precision experiment were 1.10%,0.60% and 1.10% respectively,the RSD values of daytime precision RSD were 1.31%,0.97% and 1.31% respectively,the detection limit was 0.025 μg/mL.The optimum determination condition of the probe in vitro recovery was as followed: perfusion flowrate was 1μL/min,temperature was 37℃,and time was 9 h.(2)The chiral high performance liquid chromatography(HPLC)method for the determination of the content of ofloxacin in mice blood and organizations was established.From 0.1μg/mL to 12.8μg/mL,the content of levofloxacin and deoxyfloxacin and peak area all showed good linear relationship(r=0.9995 and r=0.9997,respectively).The RSD values of precision experimental were less than 15%,the average extraction recovery values of levofloxacin and deoxyfloxacin were both 85.16% ~94.12%,and mouse plasma and blank reagent blank were without interferenceat its peak position.(3)Based on charge transfer reaction can be enhanced with the principle of fluorescence intensity on dialysis combined with μSIA-LOV techniques ofloxacin content detection method is established.Combined with LOV multifunction flow pool in the system module,ofloxacin solution dosage of 50μL chloranilic acid solution of optimal concentration 20μg/mL and dosage of 50μL detection velocity of 20μL/s,delay time 0s,concentrations of ofloxacin in 5μg/m L~30μg/mL good linear relation with the linear range(r=0.9994),precision between 2.62%~4.73%,the detection limit is 2μg/m L.Conclusion: In this study,three methods for the determination of ofloxacin in biological samples were established.The method has good specificity and was in accordance with the requirements of biological sample analysis.It provided a reference basis for the clinical drug and on-line detection of ofloxacin. |
PDF Full Text Request