Acquisition of a Triple Stage Quadrupole Mass Spectrometer

获得三级四极杆质谱仪

• 批准号: 9414018
• 负责人: Michael Thomas
• 金额: $ 23.7万
• 依托单位: Wake Forest University School of Medicine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-10-01 至 1996-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9414018&HistoricalAwards=false
• 关键词: Acquisition; Triple; Stage; Quadrupole; Mass

This application is a request for monies that wiU be applied toward the purchase of modern MS/MS instrumentation. Thirteen projects are included in the application. The projects have been submitted by participating investigators that are located in 7 academic departments from both the main campus and the medical center. The university-wide participation demonstrates a widespread appreciation of the critical need for in-house MS/MS at Wake Forest University. AU of the investigators have used mass spectrometry in their research and the availability of MS/MS techniques wiU dramatically improve the effectiveness of these investigators. The MS/MS instrumentation will be used for the analysis of a variety of complex molecules. Samples will be ionized by one of several techniques including: electrospray, cesium ion LSIMS, or EUCI. A variety of compounds will be examined using the MS/MS instrument including small, complex organic monomers, peptides, and proteins. The VG Quattro-II triple stage quadrupole mass spectrometer having the foUowing options wiU fiU our current needs: extended mass range, EI and CI ionization source and interface, a data system controUed gas chromatograph equipped with split/splitless and cool on-column GC injectors, cesium ion LSIMS, and an API LC-MS interface for both classical and pneumaticaUy assisted low flow rate electrospray. The types of projects that will be supported by the MS/MS system fall into three broad categories. The first category of projects involves structure analysis of smaller molecules, 2,500 amu. Several projects are included from synthetic chemists desiring to determine the structures of complex organic molecules and from biochemists studying the oxidations of lipids and proteins. These investigators wiU employ LSIMS, electrospray, and El/CI techniques. The second category of projects are from investigators that need to 1) determine the masses of proteins that are then generated by site-directed mutagenesis, 2) assess modifications to proteins subjected to oxidative stress, or 3) analyze chemical modifications that are generated in mechanistic studies of enzyme action. In projects utilizing site- directed mutagenesis, it is desirable to generate proteins that have well defined modifications of the primary structure. Electrospray techniques wiU be employed to ensure that these proteins have been properly processed. In the studies on protein oxidation, the degree of oxidation and the number of oxygen atoms added to the protein will be assessed by the electrospray technique. In the mechanistic studies, reactive substrates are added to the protein and the result of the experiment is assessed by determining the change in mass of enzyme exposed to the reactive substrate. In addition to analyzing purified proteins, mixtures of peptides can be separated on the HPLC system and directly introduced into the mass spectrometer via the electrospray interface. Analysis of these fragments is essential for determining the specific region of the protein that has undergone chemical changes. The last category of projects uses MS/MS for two types of analyses. The first category will employ CID techniques to assess the fatty acid composition of phospholipids. One of the users will employ this technique to identify which phospholipids contain arachidonic acid and then assess how the distribution of arachidonic acid changes in response to the activation of metabolic pathways that utilize arachidonic acid. The second type of analysis will use CID techniques to determine the sequence of small peptides. This type of analysis will be employed to verify the site of substitution by mutagenesis, determine the primary sequence of a peptide, or determine which amino acid has been modified after free radical oxidation or after other chemical treatments. At least two significant benefits will accrue to the university from the acquisition of MS/MS instrumentation. The first benefit is an improved research environment. Eight of the thirteen investigators are junior faculty less than 40 years of age and access to MS/MS is essential for their career development. The programs of senior investigators will continue to grow by having access to advanced MS/MS techniques. The second benefit is the improved training that will be available to our graduate students and postdoctoral fellows. Page 4

该申请请求资金用于购买现代 MS/MS 仪器。申请中包含十三个项目。这些项目是由来自主校区和医疗中心的 7 个学术部门的参与研究人员提交的。全校范围内的参与表明了维克森林大学对内部 MS/MS 的迫切需求的广泛认识。所有研究人员都在他们的研究中使用了质谱分析，MS/MS 技术的可用性将极大地提高这些研究人员的效率。 MS/MS 仪器将用于分析各种复杂分子。 样品将通过多种技术之一进行电离，包括：电喷雾、铯离子 LSIMS 或 EUCI。使用 MS/MS 仪器可以检测多种化合物，包括小型、复杂的有机单体、肽和蛋白质。 VG Quattro-II 三级四极杆质谱仪具有以下选项，可满足我们当前的需求：扩展的质量范围、EI 和 CI 电离源和接口、配备分流/不分流和冷柱头 GC 进样器的数据系统控制气相色谱仪、铯离子 LSIMS 以及用于经典和气动的 API LC-MS 接口 辅助低流速电喷雾。 MS/MS 系统支持的项目类型分为三大类。第一类项目涉及较小分子（2,500 amu）的结构分析。其中包括一些来自希望确定复杂有机分子结构的合成化学家和研究脂质和蛋白质氧化的生物化学家的项目。 这些研究人员将采用 LSIMS、电喷雾和 El/CI 技术。 第二类项目来自研究人员，他们需要 1) 确定通过定点诱变产生的蛋白质质量，2) 评估氧化应激下蛋白质的修饰，或 3) 分析酶作用机制研究中产生的化学修饰。在利用定点诱变的项目中，需要产生具有明确的一级结构修饰的蛋白质。将采用电喷雾技术来确保这些蛋白质得到正确处理。在蛋白质氧化的研究中，氧化程度和添加到蛋白质中的氧原子数量将通过电喷雾技术进行评估。在机理研究中，将反应性底物添加到蛋白质中，并通过确定暴露于反应性底物的酶的质量变化来评估实验结果。除了分析纯化的蛋白质外，还可以在 HPLC 系统上分离肽混合物，并通过电喷雾接口直接引入质谱仪。对这些片段的分析对于确定发生化学变化的蛋白质的特定区域至关重要。 最后一类项目使用 MS/MS 进行两种类型的分析。第一类将采用 CID 技术来评估磷脂的脂肪酸组成。其中一位用户将采用这种技术来识别哪些磷脂含有花生四烯酸，然后评估花生四烯酸的分布如何响应利用花生四烯酸的代谢途径的激活而变化。第二种类型的分析将使用 CID 技术来确定小肽的序列。此类分析将用于验证诱变取代的位点，确定肽的一级序列，或确定自由基氧化或其他化学处理后哪个氨基酸已被修饰。 购买 MS/MS 仪器至少会给大学带来两个显着的好处。第一个好处是改善了研究环境。 13 名研究人员中有 8 名是年龄不到 40 岁的初级教员，获得 MS/MS 对他们的职业发展至关重要。通过使用先进的 MS/MS 技术，高级研究人员的项目将继续发展。第二个好处是我们的研究生和博士后研究员可以获得更好的培训。 第4页