Based On UHPLC-Q-TOF-MS/MS Technology To Exploration Of Endogenous Metabolites Related To Diagnosis And Treatment Of CRC And Prediction Of Adverse Reactions To Capecitabine In Blood

Objective:to investigate the endogenous metabolites related to the diagnosis and treatment of CRC and the prediction of adverse reactions of capecitabine in patients with CRC based on UHPLC-Q-TOF-MS/MS technique.Methods:in order to detect more endogenous metabolites,the blood was first treated by protein precipitation method,and then the method of liquid-liquid extraction was used.The blood samples were divided into two layers:polar metabolites in the upper layer and non-polar metabolites in the lower layer,and then determined by computer.The column used for polar metabolites in the upper layer was HILIC-Z?2.1×150 mm?column HILIC-Z?2.1×5 mm?.According to the proportion of mobile phase,pure water?10mol/L ammonium formate?-90%acetonitrile?ammonium 10mol/L formate?was eluted by gradient program,the flow rate was0.5mL·min^-1,and the column temperature was 40?.The column used for nonpolar metabolites in the lower layer was C18?2.1×100 mm,1.7?m?column:C18?2.1×5 mm,1.7?m?;According to the mobile phase ratio:60%acetonitrile aqueous solution?0.1%formic acid 10mol/L ammonium formate?-isopropanol:water=9:1(0.1%formic acid10mol/L for programmed elution,flow rate 0.3mL·min^-1,column temperature 65?.The data collected at the first level were analyzed by non-targeted metabonomics.Firstly,?PCA?analysis,orthogonal partial least square discriminant?OPLS-DA?analysis and variable weight importance ranking?VIP>1?analysis of principal components are carried out by using SIMCA software.At the same time,the characteristic ion peaks of volcanic maps were made by MetaboAnalyst software?P<0.05 and FC>2?.Finally,the characteristic ion peaks satisfying VIP>1 and P<0.05,FC>2 were screened out,and the tandem mass spectrometry data of the selected compounds satisfying the conditions were analyzed by targeted second-order analysis.The collected data of targeted secondary analysis are matched with HMDB and METALIN,respectively.Finally,the diagnostic value of selected compounds with biological significance was evaluated by SPSS 13.0 software.Results:Succinicacidsemialdehyde,N1N12Diacetylspermine,Adenosine5monophosphate,Octanoyl-CoA,Creatinine.,a potential biomarker of biological significance,was found in the upper blood of patients with colorectal cancer.Triethylamine,N-stearoyl valine,GlcCer,a potential biomarker of biological significance,was screened from the lower blood?d16:1/24:0?.It is of diagnostic significance to evaluate the blood compound Succinicacidsemialdehyde,N1,N12Diacetylspermine,Octanoyl-CoA,GlcCer?d16:1/24:0?in patients with colorectal cancer.At the same time,the compound Bis?2-hydroxyethyl?dithiocarbamic acid,GlcCer?d16:1/24:0?has the ability to predict hand and foot syndrome at the same time.Conclusion:based on UHPLC-Q-TOF-MS/MS technique,the metabolites which can be used to diagnose colorectal cancer and predict the adverse reactions of capecitabine adjuvant chemotherapy were screened out from the blood by non-targeted metabonomics.Therefore,non-targeted metabonomics can be used to detect colorectal cancer and find new potential metabolic markers,which provides the research direction and train of thought for the later clinical study of colorectal cancer and the individualized treatment of patients.