質譜技術 在 高科技產業 上的應用

1. 質譜技術 在 高科技產業 上的應用 中央研究院原子與分子科學研究所 韓肇中 cchan@po.iams.sinica.edu.tw

2. 醉月湖 原分所 公 館臺 大 校 總 區

3. Mass spectrometry is an analytical technique that identifies the chemical composition of a compound or sample based on the mass-to-charge ratio of charged particles. — Wikipedia

4. What is Mass Spectrometry? Mass spectrometry is the art of measuring atoms and molecules to determine their molecular weight. Such mass or weight information is sometimes sufficient, frequently necessary, and always useful in determining the identity of a species. To practice this art one puts charge on the molecules of interest, i.e., the analyte, then measures how the trajectories of the resulting ions respond in vacuum to various combinations of electric and magnetic fields. — John B. Fenn, Nobel Laureate in Chemistry 2002.

5. Nobel Prize winners for their advancement of Mass Spectrometry J.J. Thomson 1906 Physics Francis W. Aston 1922 Chemistry Wolfgang Paul & Hans G. Dehmelt 1989 Physics John B. Fenn & Koichi Tanaka* 2002 Chemistry * As of 2008, he is the only person without a post-bachelor's degree to have won a Nobel Prize in a scientific field.

7. Ion source — electrospray ionization (ESI) 電灑, nanoelectrospray ionization (nanoESI), matrix-assisted laser desorption/ionization (MALDI) 介質輔助雷射脫附游離, atmospheric pressure chemical ionization (APCI), corona discharge 電暈放電, desorption/ionization on silicon (DIOS), fast atom/ion bombardment (FAB/SIMS), electron ionization (EI), chemical ionization (CI).

8. V too low V too high

10. http://www.eng.qmul.ac.uk/subsites/electrospray/videos.php

12. Mass analyzer — Quadrupole mass filter (QMS) 四極矩, quadrupole ion trap (QIT, 3-D ion trap or Paul trap; 2-D or linear ion trap) 離子阱, Time-of-Flight (TOF), Magnetic sector, Fourier Transform Ion Cyclotron Resonance (FTMS, Penning trap), orbitrap, and various tandem combinations: Quad-TOF (Q-TOF), TOF-TOF, Double-focusing 雙聚焦： EB (normal geometry tandem sector instrument), BE (reversed-geometry tandem sector instrument).

13. + + + + Quadrupole mass filter (QMS) 四極矩

14. Paul Trap離子阱質譜儀之核心 剖面示意圖 實物拆解照片 Penning (ion) traps are used in the physical realization of quantum computation and quantum information processing

15. QIT, 3-D ion trap or Paul trap

17. TOF-MS http://www.kore.co.uk/1095.htm

18. Double focusing sector mass spectrometer

19. FTMS, Penning trap

20. Thermo-Fisher LTQ-FT LTQ-Orbitrap

21. Varian Bruker Daltonics

23. Detectors • Faraday Cup • Secondary Electron Multipliers (SEM) • Channel Electron Multiplier (CEM) • Multichannel Array Detector

24. Detector — Electron multiplier, photomultiplier conversion dynode Venetian blind/discrete dynode electron multiplier Continuous dynode electron multiplier

25. Organization for Economic Co-operation and Development (OECD經濟合作開發組織) classification of industries - 1973

26. High tech --- Wikipedia High tech is technology that is at the cutting edge — the most advanced technology currently available. There is no specific class of technology that is high tech—the definition shifts over time …. As of 2006, any technology from the year 2000 onward may be considered high tech. • Aerospace technology — extraterrestrial material — meteorites, interplanetary dust Biotechnology • Information technology Nanotechnology • Robotics

27. Applications of MS in semiconductor industry • Process gas analysis – eg. Contamination and composition • Adsorbed surface layer analysis • Secondary Ion Mass Spectrometry (SIMS) • Thin film/layer deposition QC • Lateral 2-D pattern (static SIMS) and 3-D depth profiling (dynamic SIMS) • Device failure diagnosis • Reverse engineering (材料逆向工程分析) • Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry

28. Surface analysis using secondary-ion mass spectrometry (SIMS). David G. Castner, Nature 422, 129-130 (2003)

31. http://images.google.com.tw/imgres?imgurl=http://www.condensed-matter.uni-tuebingen.de/resources/pictures/simsschema.jpg&imgrefurl=http://www.condensed-matter.uni-tuebingen.de/cms/parser/parser.php%3Ffile%3D/en/facilities/sims.htm&ushttp://images.google.com.tw/imgres?imgurl=http://www.condensed-matter.uni-tuebingen.de/resources/pictures/simsschema.jpg&imgrefurl=http://www.condensed-matter.uni-tuebingen.de/cms/parser/parser.php%3Ffile%3D/en/facilities/sims.htm&us

33. Ion imaging Ion-images of the surface was recorded with a lateral resolution of up to 5µm. The primary ion beam is scanned over the surface and for every point the intensity of the chosen mass is recorded. The picture on the left shows an image of a metal surface. In the right corner of the window there was a SED - image of the sample. The width of this image is about 300µm. The right picture shows the titanium impurities of a natural SnO2 single crystal surface (110). The width of this picture is about 700µm http://images.google.com.tw/imgres?imgurl=http://www.condensed-matter.uni-tuebingen.de/resources/pictures/simsschema.jpg&imgrefurl=http://www.condensed-matter.uni-tuebingen.de/cms/parser/parser.php%3Ffile%3D/en/facilities/sims.htm&us

34. Inductively Coupled Plasma Time-of-Flight (TOF) Mass Spectrometer

35. Staticic SIMS

37. Depth profiling – Dynamic SIMS Depth profile of a sample, the beam is scanned in a spiral and the counters are only open, when the beam reaches a central region of the sputter-crater. With this method it is possible to avoid edge effects and to get only a signal of a defined sputter-depth. Both profiles have been taken on the same sample (tinoxide coated silicium wafer), detecting positive (left) and negative (right) ions with a total depth of about 350nm http://images.google.com.tw/imgres?imgurl=http://www.condensed-matter.uni-tuebingen.de/resources/pictures/simsschema.jpg&imgrefurl=http://www.condensed-matter.uni-tuebingen.de/cms/parser/parser.php%3Ffile%3D/en/facilities/sims.htm&us

38. MS Applications in Biotechnology: • Genomics • Proteomics • Metabolomics • Glycomics • Lipidomics • Interactomics • Protein Biomarker identification

39. The official NIH (USA) definition of a biomarker is: "a characteristic that is objectively measured and evaluated as an indicator of normal biologic processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention."

40. Advantages of protein Markers • Proteins are the direct machinery executing biological functions much more direct indicationthan DNA’s or RNA’s of pathologic status. • Proteins with the correct amino acid sequence as mandated by a “normal” DNA can still be the cause of a disease state, as amino modifications, such as phosphorylation, glycosylation, alkylation, etc. can all affect a protein’s 3-D structures and thus functions. (Post-translational modification) • Secretory proteins can be sampled remote from thenidus, well before metastasis occurs.

41. sample collection protein separation protein identification bioinformatics cells body fluids tissues protein quantification chromatography electrophoresis ... Edman sequencing mass spectrometry Application of Proteomics in Cancer Research General Proteomic workflow

42. MudPIT (Multidimensional Protein Identification Technology) MudPIT is a technique for the separation and identification of complex protein and peptide mixtures. Rather than use traditional 2D gel electrophoresis, MudPIT separates peptides in 2D liquid chromatography. In this way, the separation can be interfaced directly with the ion source of a mass spectrometer.

43. Crude Separation Proteins from Small Molecules • Small molecules, e.g., peptides, amino acids, and metabolites, adversely influence MudPIT performance and mass spectrometric analysis. • Many methods for removal of interfering small molecules had been evaluated. Among these attempts, dialysis, various protein precipitation protocols with the addition of organic acids and/or organic additives, and molecular weight cut-off ultrafiltration, were either too laborious or too lossy. • Soluble proteins with MW > 4 kD is separated from small molecules by size exclusion (gel filtration) chromatographyfor further MudPIT-MS analyses.

44. Major Approaches for Protein Identification Science 2006, 314, 65 – 66.

47. Potential problems with bottom-up approach • A mixture of n-proteins is converted into a (10~50)xn-peptide mixture. Increase in complexity is more than compensated by gain in MS detection sensitivity. • The exact “intactness” of the proteins in the starting sample is not assured. • Quantitation of each parent protein content is almost impossible.

48. Information on the x-yth dimensional section Top-Down Bottom-Up D-y (Mass) D-y (Mass) D-x (LC retention time) D-x (LC retention time)

49. Introductory Mass Spectrometry Website by Dr. Lhoëst G.J.J. http://www.specmetcrime.com/mass__spectrometry.htm • Ionization methods in MS • APCI and ESI • FAST ATOM AND ION BOMBARDMENT • PHOTOIONIZATION • CHEMICAL IONIZATION • ELECTRON IONIZATION (EI) • Analyzers • MAGNETIC AND ELECTROMAGNETIC • TIME- OF- FLIGHT • Quadrupole Mass Filter • Tandem MS with triple quadrupole mass spectrometers • Quadrupole Ion Trap (QUIT/Paul trap) • Fourier Transform Ion Cyclotron Spectroscopy (FTICR/Penning Trap) • Detectors • Faraday Cup • Secondary Electron Multipliers (SEM) • Channel Electron Multiplier (CEM) • Multipoint Array Detector

50. The basics of mass spectrometry in the twenty-first century Gary L. Glish & Richard W. Vachet Nature Reviews Drug Discovery 2, 140-150(February 2003) a | Mass analysis in time-of-flight (TOF) spectrometry is achieved because ions of different mass-to-charge (m/z) values have different velocities and therefore reach the detector at different times. b | A double-focusing analyser provides direction focusing through both the electric and magnetic sectors. Ions with the same kinetic energy-to-charge ratio follow a common path through an electric sector, and ions can then be dispersed according to their momentum-to-charge ratio in a magnetic sector. Overall analysis according to m/z is achieved. c | In a quadrupole mass analyser(top rod not shown), the correct magnitude of the radio frequency and direct current voltages applied to the rods allows ions of a single m/z to maintain stable trajectories from the ion source to the detector, whereas ions with different m/z values are unable to maintain stable trajectories.