Chromatography

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Chromatography par Mind Map: Chromatography

1. HPLC

1.1. High Performance Liquid Chromatography

1.2. Specific Methods/Stationary Phase/Type of Equilibrium: - Liquid-liquid / Liquid adsorbed or bonded to a solid surface / Interaction with or partitioning between immiscible liquids - Liquid-solid or adsorption / Solid / Adsorption - Ion exchange / Ion exchange resin / Ion exchange - Size exclusion / Liquid in interstices of a polymeric solid / Partition or sieving - Affinity / Group-specified liquid bonded to a solid surface / Partition between surgace liquid and mobile liquid

1.3. Normal Phase: Hydrophilic stationary phase, hydrophobic mobile phase Reversed Phase: Hydrophobic stationary phase, hydrophilic mobile phase

2. Mass Spectrometry Applications

2.1. Analyte ionized and detected according to mass-to-charge ratio

2.2. Compatibility: Gas must be able to be ionized, liquids volatilized, low salt

2.2.1. Gases: Thermally stable. boiling point <500 C, molar mass < 1000 Da

2.2.2. Desorption and electrospray: Doesn't require volatilization, molar mass up to 10^5 Da

2.2.3. General ionization sources: Hard sources - molecules excited and energy dissipated through lots of fragmentation Soft sources - little fragmentation

2.2.3.1. Types of Ion Sources: - Electron ionization - Chemical Ionization - Electrospray Ionization - Atmospheric pressure chemical ionization - Matrix-assisted laser desorption/ioniation

2.2.3.2. Hard source: Sample vaporized at high temp, bombarded with beam of electrons Usually coupled with gas chromatography Many fragment ions creased Mass range <1000 Da

2.2.3.3. Soft source: High energy electrons ionize reagent gas to collide with analyte, transferring protons Most commonly electrospray and matrix-assisted laser desorption ionization

2.3. Setup: Ionizer - Mass analyzer - Detector

2.4. Mass analyzers

2.4.1. Types: Magnetic sector Quadrupole Ion Trap Time-of-Flight FT-ion cyclotron resonance Orbitrap

2.5. Detectors

2.5.1. Types: Faraday collector Electron multiplier

3. Basics

3.1. Plate theory: equilibrium between the mobile and stationary phase for each species

3.1.1. Column efficiency: Height equivalent of a theoretical plate (H=L/N)

3.2. Rate theory: relating diffusion and resistance to mass transfer

3.2.1. Eddy diffusion

3.2.2. Longitudinal diffusion

3.2.3. Resistance to mass transfer in mobile phase

3.2.4. Resistance to mass transfer in stationary phase

4. Gas Chromatography

4.1. Specific Methods/Stationary Phase/Type of Equilibrium: - Gas-liquid / Liquid adsorbed or bonded to solid surface / Partition between gas and liquid - Gas-solid / Solid / Adsorption

4.2. General layout: Flow controller - Sample injector - Column - Detector/Signal Processing - Waste

4.3. Flame Ionization Detector (FID) Eluate burned and electron produced, which is carried to the detector