| PurposePolyamines are kinds of aliphatic compounds with several amidogens.Including Putrescine(Put),Spermidine(Spd),Spermine(Spm).Polyamines are extensively existed in tissue and body fluid.Since 1971,the possibility which polyamine as clinical tumor diadynamic criteria has already caused extensive interest,a great deal of research prove that polyamines in body fluid and urine of various clinical cancer increased.The purpose is to use a feasible,effective,high accuracy experimental methods to determine polyamine in health adult,and try to make sure normal value of polyamine, and to apply in some actual cases,apply the feasibility in becoming diagnostic test evaluation indexing sign.Materials and Methods一,MaterialsHealthy object(22),come from China Medicine University.Tumor object(30), come from First Clinical Hospital Tumor Section of China Medicine University.One month ago,Above-mentioned objects had no symptoms,such as infection,external injury and epilepsy.etc.二,Methods(一)Sample collectionThe objects are forbid to eat for 4～8 hours before sampling,and forbid to a violent activity,take the empty stomach urina sanguinis,refrigerating In-40℃refrigerators. (二)Instrument condition1.Chromatograph conditionRtx-5MS capillary column(Restek company)(30m×0.25mm×0.25μm);Carrier gas:He(＞99.999%);Injection temperature 280℃;Incipient column temperature 70℃(keep 1 min),increasing temperature to 295℃by 25℃/min.(keep 10 mins); Splitless;Interface temperature 280℃.2.Spectrometer conditionEI source,temperature 200℃,SCAN mode.(三)Analysis stepTake 1 ml urine,add 2μl IS,and add 0.1 ml thick HCl in the tube,heats for 1.5 hours(100℃-105℃).Cool.adjust pH to 12.75,carry on solid extraction,then elute with derivatization liquid.Condense 30μl with Nitrogen gas.Determinate by GC/MS. The polyamine values need to be rectified by urine creatinine which are determined by UV.ResultsGC/MS is a sensitive,rapid detection technology.After statistical analysis by Microsoft Excel and SPSS,The range of Spm normal value(X±S,nmol/mg creatinine) is 22.964±s 3.316.Compare with tumor objects,the Spm value have significant deviation.If we contribute Spm range as evaluation indexing sign.to determine tumor,. the specificity is 81.8%,and the mistake diagnostic rate is 18.2%.Discussion1.Put shows best when we use 0.5 ml thick HCl,but Spd and Spm are not the same.We think that its best to choose 0.1 ml thick HCl and 100℃～105℃.2.The particle size of kieselguhr is 15-25 mesh.3.There are amino in Put,Spd,Spm,acylation can make its polarity lower,then we speed up determination.The stability of derivatives are very low,we found that the polyamine derivatives are stable when they are in butyl acetate and 24h under 4℃. 4.It is reported that urine polyamine are not stable,and also innfluented by kidney clearance rate,In the nervous system tumor patient,the concentration of polyamine all increase in blood and cerebrospinal fluid.If we determine the polyamine level in blood and cerebrospinal fluid at the same time,the study would have more sense.5.The evaluation of diagnosis index sign:If we contribute Spm normal value as evaluation indexing sign.To determine tumor,the specificity is 81.8%,and the mistake diagnostic rate is 18.2%.Because the objects are not enough,we need more objects to modify the result.When we make a conclusion,we have to collate clinic.6.Examination time:The whole processes of the examination include extraction, hydrolization,concentration,injecyion,GC/MS analysis,data calculation,making report,3 hours totally.7.Economics evaluation:materials(urine glass,test tube,sipper,columnella, kieselguhr),reagent(IS,thick HCl,NaOH),consumption of instrument(pH meter, GC/MS,carrier gas,heating element,injector,sample injector),service charge, 80.00 yuan totally.ConclusionCompare with the other biology materials,as a study target,urine is easy to get and preserve,otherwise,it also contains several biology information.Mass spectrometer is a kind of good instrument in appraisement.Chromatography is a kind of good instrument in separating.We combine These conditions together,and found that the range of Spm normal value(X±S,nmol/mg creatinine)is 22.964±s 3.316. They are far lower than that in tumor objects.And the specificity is 81.8%,and the mistake diagnostic rate is 18.2%.
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