Development And Application Research For The Determination Method Of BPs And Neurotransmitters By Liquid Chromatography Tandem Mass Spectrometry

Part one:Development and application research for the determination method of 13 kinds of BPs in water and sediment by UHPLC-MS/MSObjectives:A method for determination of bisphenol A(BPA)and twelve bisphenol analogues(BPs)combining pre-column derivatization after liquid-liquid extraction(LLE)and ultra-high performance liquid chromatography coupled to tandem mass spectrometry(UHPLC-MS/MS)was developed and validated in the river water and sediment samples.Methods:Analytes were extracted from river samples by liquid-liquid extraction(LLE).Dansyl chloride(DNSC)was selected as a derivatization reagent.Derivatization reaction conditions such as reaction temperature,reaction time,DNSC concentration and pH value were tested.All the derivertized target compounds were well separated and eluted in 10 min.The dansyl labeled compounds were analyzed by the high resolution mass spectrometer,ionization through ESI(+)mode,with confirmation and quantification by PRM scan mode.Results:The results of the method validation were sensitive and satisfactory.Linearity was assessed by using matrix-matched standard calibration and good correlation coefficients(R~2>0.99)were obtained.The limits of quantification for the analytes ranged from 0.005 to 0.02 ng/mL in river water and from 0.15 to 0.80 ng/g in sediment.Good reproducibility of the method in terms of intra-and inter-day precision was achieved,yielding relative standard deviations of less than 10.1%and 11.6%,respectively.Conclusion:The developed method showed satisfactory precision,good recoveries for almost all the tested BPs compounds,and adequate limits of detection for biological exposure monitoring and environmental monitoring.The performance of the method was verified by its application to real samples.Part two: Determination and analysis of 13 kinds of BPs exposure levels in urine and plasma of normal populationObjectives: The purpose of this study was to assess the exposure levels of the bisphenol structural analogues(BPs)in the urine and plasma samples of the normal population,including the free and the general forms of BPs,and to analyze the differences.Methods: Based on the former method of ultra high performance liquid chromatography tandem mass spectrometry for the determination of BPs,a quantitative analysis method for BPs in human urine and plasma matrix was established.The urine and plasma of 49 cases were enzymatically hydrolysated by glucuronase and sulfatase.The ultra high performance liquid chromatography tandem mass spectrometry(UHPLC-MS/MS)was used to detect the BPs exposure levels in the urine and plasma samples.The concentration of creatinine in urine samples was corrected for target BPs concentration.Results: The urine recovery rate was 82.8%-115.3%,the precision was 1.3%-11.9%,the plasma recovery rate was 81.3%-115.6,and the precision was 1.5%-11.6%.The high detection rates of BPs were BPA,BPS and DHBP.The detection rate of free BPA detected in plasma and urine were 85.4% and 87.5%;The detection rates of total BPA detected in plasma and urine were 91.7% and 95.8%,respectively.The free forms of BPS,the detection rates in plasma and urine were 75% and 79.2%,respectively.The detection rates of total BPS in plasma and urine were 87.5% and 91.7%,respectively.The concentration level of BPS in urine samples was higher than that of BPA.The average concentrations of BPS in the urine for free and total states were 2.44 ng/m L and 5.82 ng/m L respectively,while the average concentrations of BPA in the urine for free and total forms were 1.36 ng/m L and 4.75 ng/m L,respectively.The concentrations of the free and general states of BPS in plasma were 0.69 ng/m L and 2.48 ng/m L,and the free and general states of BPA were 1.24 and 3.91 ng/m L respectively,the concentration of BPA in plasma is still higher than BPS.Conclusion: The results showed that BPS had become the dominant type of BPs exposure in the urine samples of the population,both in the form of free and general state.The dominant exposure type of bisphenol structure analogues in plasma was BPA.population Part three: Analysis of the difference of plasma BPs and neurotransmitter levels between Parkinson and normalObjectives: An ultra high performance liquid chromatography tandem mass spectrometry(UHPLC-MS/MS)method for simultaneous determination of bisphenol structural analogues and neurotransmitters was established,in order to explore the difference of BPs and neurotransmitters in plasma between Parkinson and the normal population.Methods: The sample pretreatment process used methanol to extract the samples of plasma samples and use them for protein removal,simple and time-saving.Dansyl chloride(DNSC)was selected as a derivatization reagent and the Thermo Hypersill GOLD(2.1 × 100 mm,1.9 ?m)column was used for seperation.Acetonitrile and water containing 0.1% formic acid are used as mobile phase.Ionization by the positive mode electrospray ion source.The mass spectrometric parallel reaction monitoring model was used for detection.The BPs exposure levels and neurotransmitter concentrations in plasma of Parkinson's population and normal control group were measured by this method.Results: The derivatization technology can improve and promote the ionization of the analytes,improve the sensitivity of the detection method,and make a rapid and simultaneous determination of 25 compounds of different properties.The recoveries of BPs and neurotransmitters in the plasma samples were between 80.8%-109.1%,and the precision values were between 0.9%-15.6%.The plasma levels of BPF,BPS and TBBPA in Parkinson case group were higher than those in the control group.The concentrations of 5-HIAA,5-HT and DA in plasma in case group were lower than those in the control group,while the levels of 3-HK,NE and E in the plasma were higher than those in the control control group(P < 0.05).Conclusion: In this experiment,the ultra high performance liquid chromatography tandem mass spectrometry method was established for the simultaneous determination of thirteen kinds of bisphenol structure analogues(BPs)and twelve kinds of neurotransmitters,using the derivatization reagent dansyl chloride.There were significant differences in plasma BPs and neurotransmitter levels between Parkinson and normal controls.