本文選題：質(zhì)譜 + 內(nèi)部萃取電噴霧電離　； 參考：《東華理工大學(xué)》2017年碩士論文

【摘要】：質(zhì)譜技術(shù)具有靈敏度高、分析速度快、能提供分子結(jié)構(gòu)信息等特點(diǎn),在生命科學(xué)研究領(lǐng)域扮演著重要角色,常用于組織樣品分析。傳統(tǒng)地,組織樣品分析一般需經(jīng)過(guò)研磨、萃取、分離等繁雜的樣品預(yù)處理過(guò)程,導(dǎo)致耗時(shí)低效,無(wú)法滿足大量樣品高通量分析的實(shí)際需求,且不可避免地使組織樣品中的一些活性成分損失。近年來(lái),隨著新興常壓質(zhì)譜技術(shù)的發(fā)展,使得組織樣品可在無(wú)需樣品預(yù)處理的條件下進(jìn)行實(shí)時(shí)、非破壞、在線直接質(zhì)譜分析,大大提高了分析效率。本文著重介紹了一種新型常壓質(zhì)譜技術(shù),內(nèi)部萃取電噴霧電離質(zhì)譜(iEESI-MS)技術(shù)在植物組織、動(dòng)物組織和人體組織分析中的應(yīng)用研究,并簡(jiǎn)要展望了該技術(shù)在植物代謝組學(xué)、食品科學(xué)、臨床醫(yī)學(xué)等領(lǐng)域的發(fā)展趨勢(shì)�；趯�(duì)傳統(tǒng)質(zhì)譜技術(shù)在組織樣品分析中應(yīng)用研究的簡(jiǎn)要概述,本論文的主要研究?jī)?nèi)容如下:1.iEESI-MS技術(shù)在植物組織分析中的應(yīng)用研究。采用iEESI-MS技術(shù)對(duì)蘆筍新鮮莖塊和干燥蘆筍莖塊、不同納米材料培養(yǎng)綠豆芽的根、莖和子葉組織及不同產(chǎn)地臍橙內(nèi)表皮組織與果肉組織進(jìn)行了直接質(zhì)譜分析。1)對(duì)蘆筍新鮮莖塊和干燥蘆筍莖塊進(jìn)行直接分析,結(jié)合碰撞誘導(dǎo)解離(CID)實(shí)驗(yàn),鑒定了對(duì)藥物和自然產(chǎn)品研發(fā)具有重要意義的多種代表性營(yíng)養(yǎng)成分。該結(jié)果不僅為食源性植物組織樣品的快速分析提供了一種質(zhì)譜分析新方法,而且對(duì)農(nóng)業(yè)的發(fā)展具有一定的指導(dǎo)意義。2)對(duì)不同納米材料包括碲化鎘量子點(diǎn)(CdTe QDs)、納米金(Au NPs)和納米銀(Ag NPs)及水培養(yǎng)綠豆芽的根、莖和子葉組織進(jìn)行直接質(zhì)譜分析,結(jié)果表明納米材料會(huì)對(duì)綠豆芽的能量代謝過(guò)程(如糖類(lèi)物質(zhì)的轉(zhuǎn)化、分解等)產(chǎn)生一定的影響。該工作從分子層面上研究了納米材料對(duì)綠豆芽代謝產(chǎn)生的影響,為探究由納米材料引起的其它效應(yīng)如細(xì)胞凋亡或壞死等奠定了一定的基礎(chǔ)。3)對(duì)不同產(chǎn)地臍橙的內(nèi)果皮組織和果肉組織進(jìn)行直接iEESI-MS分析,結(jié)果表明臍橙果肉組織與內(nèi)果皮組織中的化學(xué)成分既存在相似性,也具有特異性。結(jié)合主成分分析(PCA),使不同產(chǎn)地臍橙之間的品質(zhì)差異直觀地表現(xiàn)了出來(lái)。這進(jìn)一步說(shuō)明iEESI-MS技術(shù)可直接獲得不同品質(zhì)臍橙在分子水平上的差異,有望為判斷其它果蔬品質(zhì)的優(yōu)劣提供參考。2.iEESI-MS技術(shù)在動(dòng)物組織分析中的應(yīng)用。采用iEESI-MS技術(shù)對(duì)不同肉質(zhì)品中痕量瘦肉精成分進(jìn)行了定性、定量分析,并研究了瘦肉精代謝對(duì)豬肉品質(zhì)的影響。1)利用iEESI-MS技術(shù)從分子層面上成功地實(shí)現(xiàn)了對(duì)不同肉質(zhì)品(包括豬肉、羊肉、牛肉等)中常見(jiàn)痕量瘦肉精成分的高通量篩查,結(jié)果表明iEESI-MS技術(shù)可快速檢測(cè)到不同肉質(zhì)品中的痕量瘦肉精成分,具有樣品耗量低、操作簡(jiǎn)便、分析速度快等優(yōu)點(diǎn),為打擊含瘦肉精的劣質(zhì)肉品及杜絕運(yùn)動(dòng)員食源性興奮劑超標(biāo)提供了新的科學(xué)工具。2)利用iEESI-MS技術(shù)和專用精準(zhǔn)定量取樣裝置,對(duì)豬肉樣品中的6種瘦肉精包括沙丁胺醇、克倫特羅、萊克多巴胺、妥布特羅、特布他林和溴布特羅進(jìn)行直接定量分析。實(shí)驗(yàn)結(jié)果表明,6種瘦肉精在0.01-1000μg/kg濃度范圍內(nèi),具有較好的對(duì)數(shù)型線性關(guān)系;檢出限(LOD)為0.002μg/kg,相對(duì)標(biāo)準(zhǔn)偏差(RSD)在3.2-11.8%之間。通過(guò)國(guó)標(biāo)方法氣相-質(zhì)譜(GC-MS)技術(shù)對(duì)iEESI-MS結(jié)果進(jìn)行驗(yàn)證,表明兩者結(jié)果接近度在91-95%之間。該工作進(jìn)一步將常壓質(zhì)譜技術(shù)對(duì)待測(cè)物進(jìn)行定量分析的能力從固態(tài)樣品的表面(以μg/cm2為單位)延伸到整體組織樣品內(nèi)部(以μg/kg為單位)。3)利用iEESI-MS技術(shù)分析了含克倫特羅、沙丁胺醇及正常豬肉在營(yíng)養(yǎng)成分之間的差別,結(jié)合相關(guān)性分析和PCA,找到了多種與豬肉品質(zhì)相關(guān)的生物標(biāo)志物分子,發(fā)現(xiàn)瘦肉精代謝可能影響了線粒體內(nèi)膜外存在的調(diào)控L-肉堿與脂肪酸結(jié)合的肉堿脂酰轉(zhuǎn)移酶I的生物活性,使脂肪酸進(jìn)入線粒體基質(zhì)的過(guò)程受到了影響,從而干擾了脂肪酸合成磷脂酰膽堿的過(guò)程。3.iEESI-MS技術(shù)在人體組織分析中的應(yīng)用研究。1)采用iEESI-MS技術(shù),利用21種不同組成的CH3OH/H2O為萃取溶劑,對(duì)肺癌癌癥組織與癌旁組織進(jìn)行直接質(zhì)譜分析,結(jié)果表明CH3OH:H2O=30:70(v/v)是適合肺癌組織樣品內(nèi)部磷脂類(lèi)物質(zhì)萃取的最佳溶劑條件。2)在前期最佳優(yōu)化實(shí)驗(yàn)條件下,對(duì)肺癌癌癥組織與癌旁組織進(jìn)行iEESI-MS分析,結(jié)果表明iEESI-MS技術(shù)可快速識(shí)別癌癥組織與癌旁組織在分子層次的差異。結(jié)合PCA找到了對(duì)區(qū)分癌癥組織與癌旁組織有顯著貢獻(xiàn)的磷脂類(lèi)物質(zhì)等生物標(biāo)志物分子,為進(jìn)一步探究磷脂類(lèi)物質(zhì)在癌癥發(fā)病機(jī)制中的作用、術(shù)中準(zhǔn)確判定癌癥邊界、確保惡性腫瘤完全切除等提供參考依據(jù)。
[Abstract]:Mass spectrometry has the characteristics of high sensitivity, fast analysis and providing information on molecular structure. It plays an important role in the field of life science. It is often used in the analysis of tissue samples. Traditionally, tissue sample analysis usually needs to be pretreated with complex samples such as grinding, extraction and separation, which results in low efficiency and can not satisfy a large number of samples. The actual demand for high flux analysis and the loss of some active components in the tissue samples is unavoidable. In recent years, with the development of the emerging atmospheric mass spectrometry technology, the tissue samples can be carried out in real time, non destructive and on-line direct qualitative analysis without sample pretreatment. This paper greatly improves the efficiency of analysis. A new type of atmospheric pressure mass spectrometry, the application of internal extraction electrospray ionization mass spectrometry (iEESI-MS) technology in plant tissue, animal tissue and human tissue analysis, is studied. The development trend of this technology in plant metabolism, food science, clinical medicine and other fields is briefly prospected. Based on the analysis of traditional mass spectrometry technology in tissue samples The main contents of this study are as follows: the main contents of this thesis are as follows: the application of 1.iEESI-MS technology in the analysis of plant tissue. Using iEESI-MS technology, the fresh stem block and dry asparagus stem block of asparagus, the root, stem and cotyledon tissue of different nanomaterials and the inner skin tissue of navel orange from different producing areas and flesh tissue are introduced. Direct mass spectrometry (.1) was used to direct analysis of fresh asparagus stem block and dry asparagus stem block. Combined with collision induced dissociation (CID) experiment, a variety of representative nutrients were identified for the research and development of drugs and natural products. This result not only provides a kind of mass spectrometric analysis for the rapid analysis of food borne plant tissue samples. The new method has a certain guiding significance for the development of agriculture.2) the direct mass spectrometric analysis of the root, stem and cotyledons of different nanomaterials, including cadmium telluride quantum dots (CdTe QDs), nanoscale gold (Au NPs) and nano silver (Ag NPs) and water culture green bean sprouts, indicates that the nanomaterials can metabolize the energy of mung bean sprouts (such as sugars). The effect of nanomaterials on the metabolism of mung bean sprouts was studied at the molecular level, and a basis for exploring other effects, such as cell apoptosis or necrosis, caused by nanomaterials, such as cell apoptosis or necrosis, was studied at the molecular level, and the direct iEES of the inner pericarp tissue and flesh tissue of navel orange from different origins was carried out. The results of I-MS analysis showed that the chemical components in the flesh and the endocarp tissues of the navel orange were both similar and specific. Combining with principal component analysis (PCA), the quality differences between navel oranges in different habitats were displayed intuitively. This further illustrates that iEESI-MS technology can directly obtain the difference in molecular level of different quality navel orange. The application of reference.2.iEESI-MS technology in animal tissue analysis was provided to judge the quality of other fruits and vegetables. IEESI-MS technology was used to determine the trace amount of lean meat in different meat quality, quantitative analysis, and study the effect of the metabolism of lean meat on the quality of pork.1) using iEESI-MS technology to be successful at the molecular level. The high throughput screening of trace lean meat ingredients in different meat products (including pork, mutton, beef and so on) was carried out. The results showed that the iEESI-MS technique could quickly detect the trace content of lean meat in different meat products. It has the advantages of low sample consumption, simple operation and fast analysis, which is to combat the inferior meat and Du with lean meat essence. A new scientific tool,.2, a new scientific tool for athlete food borne stimulants, is provided by.2) using iEESI-MS technology and special precision quantitative sampling device to direct quantitative analysis of 6 kinds of lean meat concentrate in pork samples including salbutamol, clenbuterol, ractopamine, Butero, terbutaline and Bbu Tero. The experimental results show that 6 kinds of lean meat essence In the concentration range of 0.01-1000 mu g/kg, there is a better logarithmic linear relationship; the detection limit (LOD) is 0.002 mu g/kg and the relative standard deviation (RSD) is between 3.2-11.8%. The results of iEESI-MS are verified by the national standard method gas phase mass spectrometry (GC-MS) technology, indicating that the results are close to 91-95%. This work further applies the atmospheric mass spectrometry technology. The capacity for quantitative analysis of the samples was extended from the surface of the solid sample (with mu g/cm2) to the whole tissue sample (g/kg unit).3) and the difference between the nutrients containing clenbuterol, salbutamol and normal pork was analyzed by iEESI-MS technology, and a variety of pork products were found in combination with the correlation analysis and PCA. The biomarkers of qualitative related biomarkers found that the metabolism of lean meat sperm may affect the biological activity of the extracellular matrix of L- carnitine and fatty acids, which regulates the presence of the mitochondria in the mitochondria, and affects the process of the fatty acids entering the mitochondrial matrix, which interferes with the.3.iEESI-MS technology of fatty acids in the synthesis of phosphatidylcholine. Application of.1) in human tissue analysis (.1) using iEESI-MS technology and using 21 different components of CH3OH/H2O as extraction solvent to analyze lung cancer tissues and adjacent tissues by direct mass spectrometry. The results show that CH3OH:H2O=30:70 (v/v) is the best solvent condition for the extraction of phosphor substances in lung cancer tissue samples,.2) at the earliest stage. IEESI-MS analysis of cancer tissues and para cancerous tissues in lung cancer showed that iEESI-MS technology could quickly identify the difference between cancer tissue and para cancer tissue at the molecular level. Combined with PCA, the biomarker molecules, such as phosphor substances, which have significant contributions to the differentiation of cancer tissues and adjacent tissues, were found. To explore the role of phospholipids in the pathogenesis of cancer, to provide accurate evidence for accurate diagnosis of cancer boundaries and to ensure complete resection of malignant tumors.
【學(xué)位授予單位】：東華理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：Q503

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