The Lysophospholipids In Biological Samples Were Analyzed By Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Lysophospholipids(LPLs),as a biologically active signaling molecule,play an important role in various life activities of cells.Previous studies have shown that LPLs in organisms are closely related to the occurrence and development of various diseases,especially as potential biomarkers for related diseases.Therefore,it is of great significance to study the method of measuring LPLs in biological samples for the diagnosis and prevention of clinically relevant diseases and the screening of specific biomarkers.In this study,Fourier transform ion cyclotron resonance mass spectrometry(FTICR-MS)was used to research the effective detection methods of LPLs in many different biological samples.The main experimental studies are as follows:1.A method for the determination of LPLs in biological samples by FTICR-MS was established;2.This paper established a matrix-assisted laser desorption/ionization FTICR-MS(MALDI-FTICR-MS)method for analyzing LPLs in biological samples;3.Using MALDI-FTICR mass spectrometry in situ imaging to analyze the spatial distribution of LPLs in biological slices;4.Preparation of specific and biocompatible surface-coated solid phase microextraction(SPME)probes for the detection of LPLs in living organisms and cancer tissues under open mass spectrometry.The main conclusions are as follows:(1)A method was developed for the determination of 4 lysophosphatidylcholine(LPCs)in biological samples by FTICR-MS.Biological samples were extracted with methanol-chloroform(9:1,V/V)and quantitative analysis using external standard method The results indicated that the four investigated lysophosphatidylcholine showed good linearity in the concentration ranges of 0.5?100?g/L,with correlation coefficients(r2)no less than 0.9930.The average recoveries at three spiked levels for practical samples ranged from 70.7%to 95.0%,with relative standard deviation(RSD)of 1.2%-9.8%.All of the experiment results deomonstrated our developed method was sensitivity,accuracy,repeatability,and suitable for rapid determination of LPCs in complex biological samples.(2)Matrix assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry(MALDI-FTICR-MS)analysis of LPLs in biological samples.The experimental conditions were based on using 2,5-dihydroxybenzoic acid(DHB)as the matrix,the laser energy was 30%,the laser frequency was 100 Hz,and the irradiation frequency was 100 Shots.Quantitative analysis was performed by external standard method,and LPLs showed good linearity in the concentration ranges of 0.5-100?g/L,with correlation coefficients(r~2)no less than 0.9930.And the obtained recoveries at three spiked levels ranged from 68.2%to 92.7%.This method was rapid and accurate,which suitable for the detection of LPLs in biological samples.(3)A method for in-situ imaging analysis of LPLs in biological slices by MALDI-FTICR mass spectrometry was established.5 mg/mL DHB with 0.1%trifluoroacetic acid(TFA)was selected as the matrix.Then the qualitative analysis of LPLs was carried out by MS/MS.Finally,the method was applied to the determination of various LPLs in zebrafish slices,which verified the applicability and reliability of the method.(4)We prepared a modified SPME probe,which completed lipid enrichment in living organisms and cancer tissues within 1 minute and desorption within 30 seconds.After comparison and verification with other methods,it is confirmed that the method was fast,stable,with good accuracy and high sensitivity.