【干货】TVNM应用文章集锦 - 脂质类

【一周干货】部分脂质类应用文章一览：

• 集成运用TVNM纳喷鸟枪法脂质组学和用HPLC 分离脂质法，建立脂质分析新平台

• 液滴萃取表面分析质谱法（LESA-MS）应用于比较脂质组学分析

• 用全自动芯片多通道纳喷和高容量离子阱质谱分析神经节苷脂

• 用TVNM芯片多通道纳喷高分辨质谱的多元化变量分析做自上而下的脂质组筛查

• 利用LESA-MS液滴萃取表面分析质谱实施人动脉粥样硬化斑块的比较脂质组学分析

华质君在文章开篇所用的老鼠图片，细细说起来，也是有点出处的：胖老鼠帮助研究人员发现脂质，可能预防或治疗糖尿病！

脂质，一个包括脂肪及相关分子的化学家族，会造成动脉阻塞和心脏病发作。国内外的许多知名大学教授及研究机构都被这些神秘的分子深深的吸引。质谱在这些研究当中必不可少，那么，作为新型自动纳升电喷雾离子源的 TriVersaNanoMate（芯片多通道纳喷离子源）在当中能发挥什么样的优异功能呢？

下面用一些文章来阐述研究者的发现和观点：

A. 集成运用TVNM纳喷鸟枪法脂质组学和用 HPLC 分离脂质法，建立脂质分析新平台

Integratingshotgun lipidomics and HPLC separation of lipids into a new platform for lipidanalysis

【Summary 】

· Development of alipidomics platform based on direct infusion and LC MS analysis

· LC MS analysis bynormal phase chromatography and nanoflow split to a chip-based infusiondevice

· Implementation of afraction collection strategy for indepth profiling of lipid species by directinfusion analysis

· This platform is apowerful tool for arraying the lipidome of poorly characterised organisms

Ionsource/Fraction collector:

NanoMateTriversa Sample analysis was performed by LC-coupling/fraction collection modeusing a post-column split where 50 nl/min was directed to the ESI-chip and 49.5µl/min was use for parallel fraction collection.

B. 液滴萃取表面分析质谱法（LESA-MS）应用于比较脂质组学分析

EvaluatingLiquid Extraction Surface Analysis (LESA) for Comparative Lipidomics Profiling

Professor Manuel Mayr and his group at the King’s British Heart Foundation Center of King’s College London evaluated liquid

extraction surface analysis (LESA) compared to previously published tissue extraction techniques. The TriVersa NanoMate LESA

capabilitywas employed to provide automated and efficient sampling. In addition, itallowed for easy analysis of one sample to act as its

owncontrol. The group was able to evaluate the ionization efficiency in bothnegative and positive mode, thereby increasing the amount

ofinformation obtained.

Bycomparing LESA to previously published tissue extraction technique of shotgunlipidomics, the group was able to show comparable and often times better datawithout sample preparation, thereby saving time and money.

【结论】

· Liquidextraction surface analysis (LESA) is a rapid approach to tissue analysiswithout sample preparation time and effort.

· Shotgun lipidomics provides valuable insights into the relationship within artherosclerotic plaques.

· LESA andother advanced mass spectrometry techniques can be employed for diagnostic anddrug screening applications due to their complete profiling of lipid speciesand classes.

C. 用全自动芯片多通道纳喷和高容量离子阱质谱分析神经节苷脂

Coupling of fully automated chip-based electrosprayionization to high-capacity ion trap mass spectrometer for gangliosideanalysis.

Almeida R¹, Mosoarca C, Chirita M, Udrescu V, Dinca N, Vukelić Z, Allen M, Zamfir AD.

NanoMaterobot was coupled to a high-capacity ion trap (HCT) mass spectrometer to createa system merging automatic chip-based electrospray ionization (ESI) infusion,ultrafast ion detection, and multistage sequencing at superior sensitivity. Theinterface between the NanoMate and HCT mass spectrometer consists of anin-laboratory constructed mounting device that allows adjustment of the robotposition with respect to the mass spectrometer inlet.

Fullyautomated chip-based nanoelectrospray was performed on a NanoMate BiVersa robotincorporating ESI 400 chip technology (Advion, Ithaca, NY, USA) controlled and manipulated by ChipSoft 7.1.1 software operating under a Windows system. The robot was coupled to the HCT Ultra massspectrometer via an in-laboratory made mounting system.

D. 用TVNM芯片多通道纳喷高分辨质谱的多元化变量分析做自上而下的脂质组筛查

Top-down lipidomic screens by multivariate analysisof high-resolution survey mass spectra.

Schwudke D¹, Hannich JT, Surendranath V, Grimard V, Moehring T, Burton L, Kurzchalia T,Shevchenko A.

Directprofiling of total lipid extracts on a hybrid LTQ Orbitrap mass spectrometer byhigh-resolution survey spectra clusters species of 11 major lipid classes into7 groups, which are distinguished by their sum compositions and could be identifiedby accurately determined masses. Rapid acquisition of survey spectra wasemployed as a "top-down" screening tool that, together with thecomputational method of principal component analysis, revealed pronouncedperturbations in the abundance of lipid precursors within the entire series ofexperiments. Altered lipid precursors were subsequently identified either byaccurately determined masses or by in-depth MS/MS characterization that wasperformed on the same instrument. Hence, the sensitivity, throughput androbustness of lipidomics screens were improved without compromising theaccuracy and specificity of molecular species identification. The top-downlipidomics strategy lends itself for high-throughput screens complementingongoing functional genomics efforts.

Analyseswere performed on a LTQ Orbitrap hybrid mass spectrometer (Thermo Fisher Scientific, Bremen, Germany)equipped with a robotic nanoflow ion source NanoMate HD (Advion, Ithaca, NJ) with4.1-ím nozzle diameter chip or, where specified, on a static nano ESI or ESI ion sources (both from Thermo Fischer Scientific). NanoMate HD was controlled by Chipsoft 6.4. software(Advion) and operated at the ionization voltage of 1.05 kV and gas pressure 1psi. When using the nano ESI source, the spraying voltage was adjustedindividually within the range of 1.1 -1.4 kV. Metal-coated glass needles werefrom Proxeon Biosystems (Odense M, Denmark) or MasCom Analysengera¨te ServiceGmbH (Bremen, Germany). When using an ESI source, samples were infused at theflow rate of 5 íL/min delivered by a built-in syringe pump and the sprayingvoltage of 3.8 kV was applied.

E. 利用LESA-MS液滴萃取表面分析质谱实施人动脉粥样硬化斑块的比较脂质组学分析

Stegemann C1, Drozdov I, Shalhoub J, Humphries J, Ladroue C, Didangelos A, Baumert M, Allen M, Davies AH, Monaco C, Smith A, Xu Q, Mayr M.

Achip-based robotic nanoelectrospray platform interfaced to a triple quadrupolemass spectrometer was adapted to analyze lipids in tissue sections and extractsfrom human endarterectomy specimens by shotgun lipidomics. Eighteen scans fordifferent lipid classes plus additional scans for fatty acids resulted in thedetection of 150 lipid species from 9 different classes of which 24 weredetected in endarterectomies only. Further analyses focused on plaques fromsymptomatic and asymptomatic patients and stable versus unstable regions withinthe same lesion. Polyunsaturated cholesteryl esters with long-chain fatty acidsand certain sphingomyelin species showed the greatest relative enrichment inplaques compared to plasma and formed part of a lipid signature for vulnerableand stable plaque areas in a systems-wide network analysis. In principalcomponent analyses, the combination of lipid species across different classesprovided a better separation of stable and unstable areas than individual lipidclasses.

Thiscomprehensive analysis of plaque lipids demonstrates the potential oflipidomics for unraveling the lipid heterogeneity within atheroscleroticlesions.

Liquidextraction surface analysis (LESA) coupled to nano-ESI-MS.

Frozenhuman plaques were cut at 200 μm using a rotary microtome (Microm HM560cryostat, Thermo Scientific), placed on electrostatically charged slides(Superfrost Plus, BDH) and air dried for 15–30 min. Lipids were directlyanalyzed from tissue sections with a Advion TriVersa NanoMate system (Advion Inc.,Ithaca, NY) controlled by Chipsoft software (v8.1.0.928, Advion) coupled to atriple quadrupole mass spectrometer (QqQ-MS, TSQ Vantage, ThermoFisherScientific, UK).

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