The Study And Exploration Of Exhaled Breath In The Patients Of Liver Disease

Background and objectives:The detection of exhaled breath has been paid more and more attention as a noninvasive method to explore the physiological and pathological process of human body.In the middle of 20 th century,Linus Paulin used gas chromatography technology to analyze the exhaled breath of human bodyand put forward "fingerprint" for the first time,which led to the rapid development of exhaled breath research.At present,more than 3000 volatile compounds have been detected in human exhaled breath,including alkanes,olefins,aldehydes and ketones,nitrogen compounds and sulfur compounds.Liver diseasescan be caused by many reasons in our country and the most common are viral hepatitis,including hepatitis B,hepatitis C,alcohol,schistosomiasis,drugs,autoimmune liver disease,etc.And hepatitis B has been reached 120 million in China,so the study of liver diseasesis particularly important.In this study,we have explored the exhaled breath of patients with liver diseases in Child-Pugh A,B,C and healthy volunteers by EESI-MSin preliminary.Methods:1.Gram-negative bacteria 134 patients admitted from the infectious disease department in the first affiliated Hospital of Nanchang University from June 2015 to January 2017 with liver diseases are recruited in our studyaccording to CHILD scoring criteria.The clinical data and laboratory examination were collected including age,sex,alanine aminotransferase(ALT),aspartate transaminase(AST),total bilirubin(TB),direct bilirubin(DB),Albumin(ALB),prothrombin time(PT),prothrombin activity(PTA),HBV-DNA.The patients were divided into three groups of A,B and C by Child-Pugh grading criterion,and 27 healthy volunteers were included.The exhaled breath samples of all subjects were collected.2.ESI solvent:(methanol/water,1:1 v/v)containing 10 ppm silver nitrate and acetic acid(10%v).building enclosed electrospray ionization mass spectrometry(EESI-MS)experimental device and analyzing the exhaled breath samples to obtain the mass spectrum.3.Xcalibur system,partial least square(PLS)analysis and SPSS(version 24)were used to perform all data.Then analyze the differences among healthy volunteers,patients with liver diseases in Child Pugh A,B,C.To identify the VOCs,the difference peak was analyzed by tandem MS,through the comparison ofthe results searched in the database,standard identification,and detailed investigation of MS fragmentation characteristics.And analyze the relationship between the corresponding compound and liver diseases.Results:1.PLS was used to analyze the exhaled breath samples amongliver patientsin Child Pugh A,B,C and healthy volunteers.There was no significant difference between healthy volunteers and liver patients in Child A.However,there were differences between healthy volunteers and liver patients in Child B,healthy volunteers and liver patients in Child C,liver patients in Child B and liver patients in Child C.2.There were significant differences betweenhealthy volunteers and the patients with Child-Pugh B.When the VIP threshold was 1.7 and combing theIndependentSamples T test(p < 0.05),9 valuable m/z values were selected.When the VIP threshold was 1.75 and combing theIndependent-Samples T test(p < 0.05),17 valuable m/z values for exhaled breath samples from Child-Pugh C patients versus healthy volunteers.When the VIP threshold was 1.8 and combing the IndependentSamples T test(p<0.05),5 valuable m/z values for exhaled breath samples from Child-Pugh B patients versus Child-Pugh C patients.Then we identify the possible corresponding materials from the corresponding high resolution accurate values in mass spectrum.3.Nine different m/z values differed significantly between Child-Pugh B patients and healthy subjects,and 6 metabolites were detected including isobutene(m/z 57),acetone(m/z 59),ethanol(m/z 61),2-butanone(m/z 73),cyclopentanone(m/z 85),2-ethylacrylic acid(m/z 101).Seventeen different m/z values differed significantly between Child-Pugh C patients and healthy subjects,and 12 metabolites were identified including isobutene(m/z57),acetone(m/z 59),glycolaldehyde(m/z 61),isoprene(m/z 69),2-butanone(m/z 73),cyclopentanone(m/z 85),2,5-two methyl furan(m/z 97),2-ethyl acrylic acid(m/z 101),2-methyl butyric acid(m/z 103),6-methyl-hexane-2-one(m/z 115),octyl aldehyde(m/z 129),glycolaldehyde complex(m/z 121).Five different m/z values differed significantly between Child-Pugh B patients and Child-Pugh C patients,and 3 metabolites were identified including m/z 105,m/z 175,m/z 189.They all belongs to glycolaldehyde complex.Conclusions:1.PLS can distinguish the exhaled breath samples between healthy volunteers and Child-Pugh B patients,healthy volunteers and Child-Pugh C patients,Child-Pugh B patients and Child-Pugh C patients.And there were differences among three groups.2.We have identified 15 valuable metabolites including isobutene(m/z 57),acetone(m/z 59),ethanol(m/z 61),isoprene(m/z 69),2-butanone(m/z 73),cyclopentanone(m/z 85),2,5-two methyl furan(m/z 97),2-ethylacrylic acid(m/z 101),2-methyl butyric acid(m/z 103),6-methyl-hexane-2-one(m/z 115),octyl aldehyde(m/z 129),glycolaldehyde complex(m/z 105,m/z 121,m/z 175,m/z 189).The relationship between the above compounds and liver disease can be preliminarily inferred.However,this paper is only a preliminary study of exhaled breath samples for liver diseasesand the significance of these VOCsare not clear.