Types of Analyzers: Quadrupole: mass filter -part1
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1 16 Types of Analyzers: Sector or double focusing: magnetic and electric Time-of-flight (TOF) Quadrupole (mass filter) Linear ion trap Quadrupole Ion Trap (3D trap) FTICR fourier transform ion cyclotron resonance (magnet) Orbitrap 17 Quadrupole: mass filter -part1 Mass filter separation accomplished using combination of DC and RF electric fields. Ions oscillate through filter towards detector. Only ions with stable trajectory transmitted. Scan DC/RF m/z. DC = Direct current (constant) RF=AC= alternating current 1
2 18 Quadrupole: mass filter -part2 The AC field makes the ions go off into spirals as they pass down the quadrupole. The DC drags them in one constant direction, towards one pair of electrodes. A small ion will be dragged a large distance by the AC, and will find itself in stronger and stronger regions of field. It will quickly collide with an electrode and disappear. A very large ion will not be affected much by the AC, but will gradually drift in the DC part. The AC is not strong enough to drag it back as it wanders, so it also collides with an electrode, and is lost. An ion that is the right size drifts slightly in the DC of the field, but is always dragged back by the AC. The AC, however, is not quite strong enough to make it spiral out of control into an electrode. Thus an ion just the right size is stable in this quadrupole field and reaches the end, where it can be measured. The stability of an ion depends on the sizes of the AC and DC ratio Quadrupole: mass filter -part3 Equations of ion motion Mathieu functions DC = Direct current (constant) = U RF=AC= alternating current = V 2
3 20 Quadrupole: mass filter -part3 scan Scan: changes of DC and AC at DC/AC constant ratio Scan-line passes through the stable region ion will pass through the quadrupole to the detector 21 Quadrupole: mass filter -part4 resolving power Unit resolution instruments : R= 20 at m/z 20; R= 200 at m/z 200 Mass range m/z aprox 3000 Easy to connect with Chromatography Techniques 3
4 22 Quadrupole: AC only (RF-only)= ION GUIDE Also HEXAPOLES, OCTOPOLES = more poles leads to better ion guiding and wider m/z range Use to adapt Atmospheric Pressure ion sources to mass analyzers acting as pipes and fully permeable to neutrals. However, the presence of residual gas reduce the ion kinetic energy Favoring also the ion motion towards the central axis of the quadrupole = Collision Cooling Ion storage device Ion Trap ion TRAPPING.exe Linear ion trap 23 Linear Ion trap : quadrupole+ trapping potential well Axial or Radial ejection of the ions Progressively ejected ions by increasing RF voltage (AC) parent ion isolation.exe LIT are used to collect ions before injecting them in bunches into other analyzer (TOF or FTIR) 4
5 24 Linear Ion trap and Quadrupole: Benefits. Good reproducibility. Relatively small and low-cost systems. Low-energy collision-induced dissociation (CID) MS/MS spectra Limitations. Limited resolution. Peak heights variable as a function of mass (mass discrimination). Peak height vs. mass response must be 'tuned'.. Not well suited for pulsed ionization methods. Low-energy collision-induced dissociation (CID) MS/MS depends strongly on energy, collision gas, pressure, and other factors. Applications. Majority of benchtop GC/MS and LC/MS systems. Triple quadrupole MS/MS systems. Hybrid MS/MS systems Example : Target analysis in drug detection 25 parent ion isolation.exe ion TRAPPING.exe 5
6 26 3D Ion trap : Mathieu Stability diagram Ion motion 27 3D Ion trap : MS n = SEVERAL fragmentation (CID) RF set so that all ions above specific m/z follow stable trajectory, All other ions escape. parent ion isolation.exe Fragmentation by collision induce dissociation (CID) : Ion activated by collision and allowed to dissociate CID = ion accelerated by electric potential and allowed to collide with neutral molecules = kinetic energy is transformed into internal energy = breakage CID depends strongly on energy, collision gas, pressure, and other factors = no libraries 6
7 28 3D Ion trap : Example D Ion trap : Fast scanning at unit resolution High resolution over smaller m/z ranges MS n exsperiments Compact = IDEAL FOR FIELD APPLICATIONS MIMS Membrane introduction mass spectrometry Aqueous Monitoring/sampling Method for airborne and aqueous organics analysis Mass spectrometry detection (MS/MS), EI, Ion Trap Morten Martinsen, Rudolf Schmid Institutt for Kjemi, NTNU, Norway 7
8 30 3D Ion trap : Benefits. High sensitivity. Multi-stage mass spectrometry (analogous to FTICR experiments). Compact mass analyzer (Field detector). MS n Limitations. Very poor dynamic range (can sometimes be compensated for by using auto-ranging). Subject to space charge effects and ion molecule reactions. Collision energy not well-defined MS/MS depends strongly on energy, collision gas, pressure, and other factors Applications. Benchtop GC/MS, LC/MS and MS/MS systems. Target compound screening 31 FT-ICR : Fourier transform ion cyclotron resonance Ion in a magnetic field experience Lorentz force and bend them in a circular path In ICR it is detected the frequency that the ions go around Mass-selective excitation achieve by a transverse electric field alternating 8
9 32 FT-ICR : High resolving power and mass accuracy High resolving power: Broad band detection for all frequencies in parallel FT (FID) m/z frequency bandwidth 3MHz-300KHz For a R: in m/z 500 frequency detection = (300KHz/100000)= 3Hz High mass accuracy : Elongated recorder FID allows for extremely precise determination of all cyclotron frequencies 33 FT-ICR : Benefits Highest recorded mass resolution of all mass spectrometers and High accuracy ( <1ppm) Wide mass range without lost of sensitivity Non-destructive ion detection= ion re-measurement Stable mass calibration Limitations: Needs Low Background pressure to avoid ion molecule reactions and ion neutral collisions Need high magnetic field Large and really expensive Applications : Complex mixture, e.g. crude oil ( Macromolecules Metabolomics Proteomics ( after protein digestion - fragments determination).etc.. 9
10 34 Orbitrap: Ions are trapped in Axial oscillation in a homogenous electrostatic field Helps to capture and cooling the ions 35 LTQ-Orbitrap 1. Ions are stored in the Linear Trap 2.. are axially ejected 3.. and trapped in the C-trap 4.. they are squeezed into a small cloud and injected into the Orbitrap 5.. where they are electrostatically trapped, while rotating around the central electrode and performing axial oscillation The oscillating ions induce an image current into the two outer halves of the orbitrap, which can be detected using a differential amplifier Ions of only one mass generate a sine wave signal 10
11 36 LTQ-Orbitrap CID Collision-Induced Dissociation PQD Pulsed-Q Dissociation HCD - Higher Energy Collisional Dissociation 37 LTQ-Orbitrap Benefits Parallel acquisition mode available : highly resolved and accurate analysis MS 3 fragmentation Polarity switch in the same run (+ to -) Slower Drop in resolution with increasing mass/z Limitations : Requires Ultra high vacuum Mass resolution and accuracy are linearly proportional to signal acquisition time Applications: 11
12 38 Hybrid and tandem analysers : MS/MS Why need MS/MS? -The detection of a specific compound in a mixture -Quantitation of a specific compound in a mixture -Structural determination of a compound Most of the instruments available in the market are hybrid intruments LTQ-Orbitrap FT-ICR QTOF Triple quadrouple instruments Q-TOF-ion mobility 39 Tripel-QUAD: Mode 1: Product ion scan Precursor Ions are selected and isolated Collision-Induced-Dissociation Results in fragmentation Product Ions are characterized with the second mass analyzer PRECURSOR ION SELECTION PRODUCT ION DETECTION 12
13 40 Mode 2 Mode 3 Precursor ion scan Q3 is held to measure the occurrence of a particular fragment ion, Q1 is scanned. This results is a spectrum of precursor ions that result in that particular product ion Neutral loss scan Q1 is scanned as above, Q3 is also scanned at a mass off-set to produce a spectrum of precursor ions that undergo a particular neutral loss. 41 Mode 4 Selected reaction monitoring (SRM or also called MRM) Q1 and Q3 are static; user-selected specific ions are transmitted through the first analyser and user-selected specific fragments arising from these ions are measured by the second analyser. The compound must be known and characterised in advance. BEST FOR QUATIFICATION METHODS 13
14 42 Comparison of Mass analysers 43 Courtesy of EPSRC National Mass Spectrometry Service Center 14
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