Mass Spectrometry (MS)
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1 Mass Spectrometry (MS) Alternative names: Mass spectrometric (selective) detector (MSD) Spectrometry - methods based on interaction of matter and radiation Mass spectrometry - method based on formation of ions followed by interaction of ions with electric/magnetic fields
2 MS principle and outputs of measurements 1. Formation of ions (ionization) 2. Filtration of ions (mass analysis) 3. Measuring of ions abundance in dependence on m/z value m - relative molecular weight of ion z - charge of ion (1, 2, 3, 4, 5, 6...) 4. Mass spectrum * axis x corresponds to m/z value * axis y corresponds to abundance of ions (signal intensity) J. Poustka, VŠCHT Praha, ÚAPV 2016,
3 Mass spectrum - format Graphic (profiles X lines) Tabulated m / z Intenzita Relativní intenzita (%) , , , , , , , , , , , ,65
4 Application possibilities Identification of compounds Empirical formula almost absolutely, if sufficiently high resolution is applied Identification of structure in dependence on ionization type (fragmentation) Comparison of spectra - identification based on characteristic formation of ions with specific m/z values and abundances (spectral libraries) Quantification of compounds Universal - LOD/LOQ differ according to ability to be ionized
5 Historical development of MS - summary 110 years From cathode tube to modern mass spectrometer 2010 GC-MS *1956 LC-MS(ESI) *1984 TOF-MS *1946 Development of spectra recording
6 IONS - formation and types * Various mechanisms - fundamental effect on spectrum * Ions charge - positive or negative (only one type at the time) positive or negative ionization - switching of modes * Regarding to original structure are distinguished: a) molecular ions: [M] + or [M] - b) pseudomolecular and adduct ions: [M+H] +, [M-H] -, [M+CH 4 ] + c) fragment ions: e.g. [M-CH 3 ] + J. Poustka, VŠCHT Praha, ÚAPV 2016,
7 Ionization in gas phase (a) (at reduced pressure - vacuum 10-4 to -8 torr) Electron Ionization - EI * loss of an electron after the Electron Impact or otherwise after the release of valence electrons - formation of ions: M + e - (70eV) M e - * standard EI spectra (library) - ionization energy 70 ev (1 ev = 96,487 kj/mol = 23,06 kcal/mol) * "hard" ionization a) spectrum rich of fragment ions b) molecular ions - sometimes are not stable (not present in spectrum) * applicable for: finding of empirical formula, structure analysis, quantification * application: fundamental for GC-MS, probe, moving belt, particle beam J. Poustka, VŠCHT Praha, ÚAPV 2016,
8 J. Poustka, VŠCHT Praha, ÚAPV 2016, Ionization in gas phase (a) (at reduced pressure - vacuum 10-4 to -8 torr)
9 J. Poustka, VŠCHT Praha, ÚAPV 2016, Ionization in gas phase (b) (at reduced pressure - vacuum 10-4 to -8 torr) Field Ionization - FI * loss of an electron after the passing through the high voltage electrical field ( 5 kv) - formation of ions * "soft" ionization molecular and adduct ions are dominating ([M+Na] + etc.) * applicable for: finding of empirical formula; quantification * application: additional for GC-MS, probe
10 J. Poustka, VŠCHT Praha, ÚAPV 2016, Ionization in gas and liquid phase (at reduced or atmospheric pressure) Chemical ionization - CI * adduction of reagent or capture of low energy (thermal electron) - formation of (PCI) or (NCI) ions * various conditions - no standard spectra (no library availability) * "soft" ionization only most stable ions (usually molecular), fragment ions are mostly absent in source fragmentation (loss of molecular ions) * applicable for: finding of empirical formula; quantification * application: GC-MS, LC-MS (APCI), probe, moving belt, particle beam
11 Ionization in liquid and solid phase (a) (at reduced pressure) Ionization by accelerated ions or atoms a) fast ion bombardment FIB, e.g. Cs + b) fast atom bombardment FAB, e.g. Xe, Ar * suitable for high molecular compounds * applicable for: finding of empirical formula; quantification * application: probe, continuous flow (CF FIB / FAB), moving belt / wire, LC-MS
12 Ionization in liquid and solid phase (a) (at reduced pressure) CF FIB / FAB Moving belt / wire
13 J. Poustka, VŠCHT Praha, ÚAPV 2016, Particle beam
14 Ionization in liquid and solid phase (b) (at reduced pressure) Desorption ionization a) chemical (DCI): Pt wire wetted by sample - fast heating b) plasma ( 252 Cf PD): californium decays into radioactive isotopes - formation of adducts c) laser (LDI): pulse evaporation by laser d) laser at matrix assistance (MALDI): LDI with matrix e) field (FD): ionization direct from emitter (see field ionization - FI) * structure analysis * application: probe
15 Ionization in liquid and solid phase (b) (at reduced pressure) MALDI
16 Ionization in liquid phase (c) (at reduced pressure) Thermospray - TSP * Ionization by fast evaporation at elevated temperature ( 300 C) at simultaneous nebulization to evacuated space * "soft" ionization predominant molecular and pseudomolecular ions * applicable for finding of empirical formula * application: LC-MS (first interface similar to API)
17 Ionization in liquid phase (c) (at reduced pressure) [M+H] + [M+NH 4 ] + [M-H] - (not [M + zh] z + ) t C flow rate < 1 ml/min
18 Ionization in liquid phase (a) (atmospheric pressure ionization - API) Electrospray - ESP, Electrospray ionization - ESI * Ionization by high voltage ( 5 kv) at simultaneous nebulization * "soft" ionization predominant molecular and pseudomolecular ions single and/or multicharged ions * applicable for: finding of empirical formula; quantification * application: LC-MS
19 Ionization in liquid phase (a) (atmospheric pressure ionization - API)
20 Ionization in liquid phase (a) (atmospheric pressure ionization - API) ESI [M+H] + [M+NH 4 ] + [M-H] - [M ± zh] z ± t N C 2-8 kv Flow rate ml/min Volatile modifiers: ammonium acetate, formic acid Nonvolatile modifiers: phosphate buffers
21 Ionization in liquid phase (a) (atmospheric pressure ionization - API) Types of electrospray Ionspray (ISP) - pneumatic nebulization Ultraspray (USP) - ultrasound nebulization Turbospray (TBSP) - thermal nebulization
22 Ionization in liquid phase (b) (atmospheric pressure ionization - API) Atmospheric pressure chemical ionization - (APCI) * ionization based on combination of evaporation at high temperature ( 400 C) and electric discharge ( 5 kv) at simultaneous nebulization * "soft" ionization predominant molecular and pseudomolecular ions * applicable for: finding of empirical formula; quantification * application: LC-MS
23 Ionization in liquid phase (b) (atmospheric pressure ionization - API) APCI [M+H] + [M+NH 4 ] + [M-H] - ([M ± zh] z ± ) t C Discharge: 2-8 kv Flow rate ml/min Volatile modifiers: ammonium acetate, formic acid Nonvolatile modifiers: phosphate buffers
24 Ionization in liquid phase (c) (atmospheric pressure ionization - API) Atmospheric pressure photo-ionization - (APPI) * ionization based on combination of evaporation at high temperature ( 400 C) and UV radiation ( 10 ev) at simultaneous nebulization * "soft" ionization predominant molecular and pseudomolecular ions * applicable for: finding of empirical formula; quantification * application: LC-MS
![([M ± zh] z ± ) t 200 650 C](/docs-images/84/91015360/images/25-4.jpg "UV radiation 10 ev Flow")
25 Ionization in liquid phase (c) (atmospheric pressure ionization - API) APPI [M+H] + [M+NH 4 ] + [M-H] - ([M ± zh] z ± ) t C UV radiation 10 ev Flow rate: ml/min Dopant - ionization support: toluene, benzene
26 Ionization in liquid phase (d) (atmospheric pressure ionization - API) DESI Desorption ESI
27 Ionization in liquid phase (d) (atmospheric pressure ionization - API) DART Direct Analysis in Real Time
28 Spray techniques Z-spray
29 Molecular weight Applicability of ionization techniques J. Poustka, VŠCHT Praha, ÚAPV 2016, ESI APPI APCI Particle Beam Thermospray FIB FAB GC/MS (EI/CI/FI) Nonpolar Analyte polarity Polar
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