2. Magnetic sector Ion trap MASS Analyzer Ion trap analyser. Ions are either created within or rapidly infected into the trap. The electrostatic field forces them into specific orbits, depending on the m/z ratio. By varying this electrostatic field, ions may be selectively ejected in order of increasing m/z from the trap and detected

3. MASS Spectrometry Physical Basis Differential deflection of charged species in a magnetic field. Two species, of identical charge and velocity but different mass, will be deflected differently by a magnetic field. The species with the smallest momentum,  is deflected to a greater extent than that with larger 

4. Composition of MASS Spectrometry Turbo pumps Diffusion pumps Rough pumps Rotary pumps Sample plate Target HPLC GC Solids probe DCI MALDI Electro-spray (IonSpray) APCI FAB LSIMS EI CI TOF Quadrupole Ion Trap Magnetic Sector FTMS Micro-channel Plate Electron Multiplier “Hybrid” PC’s Sun SPARKStation DECStation Inlet Ion source Mass Filter Detector Data System High Vacuum System

5. Mass spectrometry Ionizer Gas Phase Sample + _ Mass Analyzer Detector MALDI Electro-Spray Ionization (ESI) Time-Of-Flight (TOF) Quadrapole Ion-Trap Electron Multiplier (EM)

8. MALDI-TOF 질량분석기 l MALDI l Matrix Assisted Laser Desorption Ionization l TOFMS l Time Of Flight Mass Spectrometry Voyager DE-STR 337 nm UV laser MALDI cyano-hydroxy cinnamic acid

9. MALDI-TOF MS - 고분자 합성시료 및 생체물질 ( 단백질, 핵산, 탄수화물등 ) 분자량 측정 - 단백질 분자량 측정을 통한 post translational modifications 확인, recombinant protein 의 QC 등 - 합성 펩타이드의 확인, 단백질 분해를 통한 서열분석 및 단백질 ID - 핵산의 분자량 측정을 통한 서열분석, 합성확인 및 SNP 결정 - 핵산 및 단백질 분해에 의한 서열분석 - 미량 생화학, 석유화학 혼합물의 분자량 측정 - SPR bio chip 을 이용한 분자 상호작용 연구

11. MALDI Matrix-assisted laser desorption ionisation (MALDI). A pulse of laser light (UV) is directed at the sample which is Co-crystallised with a suitable matrix component. This matrix component absorbs most of the energy of the laser. Both matrix and sample are vaporised by the laser. In the gas phase, proton exchange occurs between matrix and Sample components producing MH + ions ESI Electrospary ionisation (ESI). Sample is converted to a fine spray of droplets. Matrix is evaporated from this, leading to Coulombic explosion and the liberation of ions which then are directed to the analyser and detetor. In ion spray ionisation (ISI), the sample is surrounded by a sheath of gas such as N 2 which imparts greater energy and hence facilitates faster input of sample to the system

13. TOF 에서 분자 이온들의 분리 KE=1/2mv 2 Flight Tube (0.5 – 4m) Detector Ion Source (4 – 25 KV)

15. Reflector TOF 내 분자이온의 비행 TOF (Time-of-Flight) Reflector 또는 ion mirror 는 이온발생 부위에서 동일한 질량의 이온들이 초기 에너지 분포 (initial energy spread) 를 갖는 것을 보정하여 분리능을 증가 시킨다. Flight Tube Detector Ion Source 4-25 kV Reflector (Ion Mirror)

16. Time-of-Flight (TOF) amp comp L+L+ M+M+ H+H+ +20 kV +22 kV 0 kV 1) Ions enter source region, accelerated toward reflectron. 2) Ions separate in space based on their relative mass-to-charge (m/z). 3) Ions reverse path in reflectron. 4) Ions impact detector. Flight time Signal [Molecular weight]

17. Quadrupole Quadropole mass analyser. Varying the radiofrequency (RF) component of voltage allows ‘ tuning ’ for specific m/z ratios. Ions with either larger of smaller m/z collide with rods and are lost

19. Q-TOF mass spectrometer

31. Proposed Folding Pathway of Monellin NID

32. [GdnHCl] (M) 012345 Relative Signl 0.4 0.6 0.8 1.0 0 100 200 300 400 500 ANS Fluorescence (483nm) GdnHCl Induced Denaturation of SCM 1.01.11.21.31.41.51.61.71.8 Elution Volume(V e, ml) U I N 0 M 0.8 M 1.5 M 2.0 M 2.4 M 3.4 M 5.0 M Sung et al. (2002) J. Biol. Chem.

33. [H, 15N] correlation spectra: Finger-print region for protein 11515

34. NC H D H +

35. DenaturedFolded NC H H

36. NC H H Monitor Folding Intermediate

37. Monitor Folding Stage