Research On HPLC-MS New Methods For The Detection Of Drug Toxic Substances And Human Metabolic Small Molecules

High performance liquid chromatography(HPLC)has now become one of the most commonly used methods of separation,detection and analysis.It has the advantages of high separation efficiency,high sensitivity,high degree of automation,and wide range of applications.Mass spectrometry(MS),an instrument with high selectivity and high sensitivity,when combined with HPLC,HPLC-MS combines the advantages of HPLC and MS,which can be used in drug monitoring,environmental analysis,food safety,biomedicine,natural product analysis and many other fields,especially in qualitative and quantitative aspects.HPLC-MS has been more and more widely used.Based on the previous works,focusing on the application of HPLC-HRMS in the separation and detection of small molecules,this dissertation is divided into the following five chapters:Chapter 1: The HPLC-MS methods and its applications are briefly summarized.The detection significance and determination methods of ginkgolic acids,purine compounds,and deoxynucleoside compounds are introduced.Chapter 2: Ginkgo acids(GAs)are the main toxic substances in ginkgo biloba leaves.For ginkgo biloba extractracts and its preparations,it’s important to control the content of GAs.In this work,the scanning mode of parallel reaction monitoring(PRM)was adopts.A simple,rapid,and highly sensitive HPLC-HRMS method was established for the detection of 5 ginkgolic acids in Shuxuening injection.Under the optimal conditions,the analytes were detected within 11 minutes.This method has good reproducibility and high sensitivity.The range of limits of detection(LODs)for each analyte was 0.005-0.020 μg/L.And the range of limits of quantitation(LOQs)was 5-10 μg/L.This method can be directly used to detect whether the total content of GAs in Shuxuening injection exceeds the standard,and it provides a reliable basis for the control of the quality standard of Shuxuening injection.Chapter 3: In order to reveal the relationship between purine metabolites and gout and diabetes,this work adopted the Full-MS scanning mode to establish an efficient and rapid HPLC-HRMS method for the detection of 7 purine compounds.Under the optimal conditions,the analytes were detected within 8 minutes.This method has high sensitivity.The range of LODs for each analyte was 0.001-2 n M.And the range of LOQs was 0.1-5 n M.This method has been used for the determination of purine metabolites in human serum of normal group,hyperuric acid group and hyperglycemia group,providing a basis for the clinical diagnosis of gout,diabetes and other related diseases.Chapter 4: DNA oxidative damage products have important potential value in clinical diagnosis.In this work,the PRM mode was adopted to establish the HPLC-HRMS method and the HPCE-HRMS method for the simultaneous detection of 6 deoxynucleoside compounds.The experimental conditions of the two methods were systematically optimized,and the analytical performance of the two methods were compared in terms of retention time,migration sequence,sensitivity,and reproducibility.The two methods established in this work,especially the more sensitive HPCE-HRMS method,provide a better choice for evaluating the relationship between DNA oxidative stress metabolites and diseases.Chapter 5: Conclusions and prospects.