MASS Analyzer Magnetic sector Ion trap analyser Ions

MASS Analyzer Magnetic sector 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

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, m, is deflected to a greater extent than that with larger m

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

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

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

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

TOF에서 분자 이온들의 분리 Flight Tube (0. 5 – 4 m) Ion Source (4 – 25 KV) Detector KE=1/2 mv 2

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

Time-of-Flight (TOF) +22 k. V 0 k. V 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. 0 k. V Signal 4) Ions impact detector. 0 k. V + + + LM H +20 k. V Flight time amp comp [Molecular weight]

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

Q-TOF mass spectrometer

Proposed Folding Pathway of Monellin N I D

Gdn. HCl Induced Denaturation of SCM U 500 5. 0 M 1. 0 Relative Signl 3. 4 M I 400 2. 4 M 0. 8 2. 0 M 300 N 0. 6 200 1. 5 M ANS Fluorescence (483 nm) 100 0. 4 1. 5 M 0. 8 M 0 0 1 2 3 4 5 [Gdn. HCl] (M) 1. 0 1. 1 1. 2 1. 3 1. 4 1. 5 1. 6 1. 7 Elution Volume(Ve, ml) Sung et al. (2002) J. Biol. Chem. 1. 8

[H, 15 N] correlation spectra: Finger-print region for protein 11 5

H N C D H +

Denatured Folded H N H C

Monitor Folding Intermediate H N H C

Monitor Folding Stage