本文選題：代謝組學(xué) + 肌酐��； 參考：《新疆大學(xué)》2016年碩士論文

【摘要】：代謝組學(xué)是研究生物體系受外部刺激或擾動(dòng)后所產(chǎn)生的內(nèi)源性代謝物整體及其變化規(guī)律的科學(xué)。血漿和尿液作為代謝組學(xué)的研究中最常用兩種體液樣本,對其所含的代表性代謝物進(jìn)行快速準(zhǔn)確分離與分析具有十分重要的理論價(jià)值和應(yīng)用前景。本研究以人體體液中的兩種(類)代表性代謝物肌酐和磷脂類化合物為研究對象,通過建立新的檢測方法,實(shí)現(xiàn)對它們的高選擇、高準(zhǔn)確性的分離與分析。此外,建立了基于液質(zhì)聯(lián)用技術(shù)的人體尿液代謝輪廓分析方法,為代謝組學(xué)研究提供新的技術(shù)手段。具體研究內(nèi)容如下:(1)基于LbL技術(shù)將PEI/PTA多層膜構(gòu)建于氧化銦導(dǎo)電玻璃片上作為化學(xué)修飾電極,采用循環(huán)伏安法,在銅離子輔助下測定不同濃度標(biāo)準(zhǔn)溶液中的肌酐含量,標(biāo)準(zhǔn)曲線的線性范圍為0.125~62.5μM,檢出限達(dá)到0.06μM,采用標(biāo)準(zhǔn)加入法得到平均回收率分別為106.7%、96.49%和99.86%(n=3)。將該電極應(yīng)用于人體尿液中的肌酐含量的檢測,然后用HPLC法對該測定結(jié)果進(jìn)行驗(yàn)證,結(jié)果無顯著性差異,說明該電極可準(zhǔn)確測定體液中的肌酐,具有良好的臨床應(yīng)用前景。(2)基于化學(xué)共沉淀反應(yīng)原理,采用“一鍋法”合成了乙二胺四乙酸(EDTA)功能化的磁性納米粒子(EDTA@Fe3O4)。將Zr(IV)螯合于磁性納米顆粒表面制得Zr(IV)-IMAC,并通過TEM、EDS等方法對其進(jìn)行表征。將Zr(IV)-IMAC用于富集四種磷脂標(biāo)準(zhǔn)品,首先建立了HPLC-ELSD同時(shí)測定的方法并對富集過程中的洗脫液進(jìn)行了優(yōu)化,發(fā)現(xiàn)在上清液和洗滌液中均未檢出,僅在洗脫液中可以檢測出四種磷脂標(biāo)準(zhǔn)品,回收率均大于94.23%,說明Zr(IV)-IMAC對磷脂具有很好的選擇性。將之用于實(shí)際血漿樣品中磷脂的富集,并用UPLC-QTOF進(jìn)行檢測,經(jīng)HMDB數(shù)據(jù)庫檢索,初步鑒定出可能的47種磷脂類化合物。(3)為了探究不同民族(漢族和維吾爾族)因素對人體尿液代謝輪廓的影響,共收集健康志愿者尿液樣本128例,維吾爾族和漢族各64例。優(yōu)化了色譜質(zhì)譜條件,建立了基于UPLC-QTOF平臺(tái)的代謝輪廓分析方法,并完成20例樣本分析,其中包括漢族和維吾爾族各10例。
[Abstract]:Metabonomics is the science of studying the whole and changing law of endogenous metabolites produced by external stimulation or disturbance of biological system. Plasma and urine are the two most commonly used body fluid samples in the study of metabolomics. The rapid and accurate separation and analysis of representative metabolites contained in plasma and urine have very important theoretical value and application prospect. In this study, two representative metabolites (creatinine and phospholipid compounds) in human body fluid were studied. A new detection method was established to separate and analyze them with high selectivity and accuracy. In addition, a method of urine metabolic profile analysis based on liquid-mass spectrometry was established, which provides a new technique for the study of metabolomics. The specific research contents are as follows: (1) based on LBL technique, PEI / PTA multilayer film was constructed on indium oxide conductive glass as chemically modified electrode, and the creatinine content in different concentration standard solution was determined by cyclic voltammetry under the assistance of copper ion. The linear range of the standard curve was 0.125 渭 m and the detection limit was 0.06 渭 M. the average recoveries of the standard addition method were 96.49% and 99.86%, respectively. The electrode was applied to the determination of creatinine in human urine, and the results were verified by HPLC. The results showed that the electrode could accurately determine creatinine in body fluid. Based on the principle of chemical coprecipitation, EDTA-EDTA-functionalized magnetic nanoparticles (EDTA-Fe3O4) were synthesized by one-pot method. ZrOIV-IMAC was prepared by chelating ZrOIV onto the surface of magnetic nanoparticles and characterized by TEM-EDS and other methods. The method of simultaneous determination of HPLC-ELSD was established for the enrichment of four phospholipid standard products by ZrAI-IV-IMAC. The eluate in the enrichment process was optimized. It was found that the eluate was not detected in the supernatant and washing liquid. Four kinds of phospholipid standard compounds can be detected only in elution solution. The recoveries of phospholipids are all over 94.230.The results show that ZrAI-IV-IMAC has good selectivity for phospholipids. It was used for the enrichment of phospholipid in actual plasma samples, and was detected by UPLC-QTOF, and searched by HMDB database. In order to explore the influence of different nationalities (Han and Uygur) on the metabolic profile of human urine, 128 urine samples were collected from healthy volunteers, 64 from Uygur and 64 from Han nationality. Based on UPLC-QTOF platform, the metabolic profile analysis method was established, and 20 samples were analyzed, including 10 Han and 10 Uygur.
【學(xué)位授予單位】：新疆大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R446.1

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