L11 - Lecture 11 Liquid Chromatography Liquid...

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Lecture 11 Liquid Chromatography
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Separation of non-volatile species or thermal labile compounds for quantification. The most widely used of all of the analytical techniques: Sensitivity – is ready adaptable to accurate quantitative determination Wide range of analytes : e.g., amino acids, protein, hydrocarbons, drugs, pesticides. ....... Early liquid chromatography was carried out in glass column i.d. 10-50mm, 50-500cm length, particle size 150- 200um, separation time: several hours, attempts to speed up the step by vacuum or pressure, but. ... The improvement of efficiency was achieved by decreasing the particle size of packing, but an instrument was needed to provide or operate at high pressure (vs old simple glass column with gravity flow of mobile phase) High performance liquid chromatography (HPLC) was originally to distinguish these newer procedures from the old method First HPLC in 1963 Liquid chromatography (LC) Column: 50-500cm length, particle size 3-10um
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GC Setup
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LC Setup
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pump degasser Auto-sampler and injector Detector Solvent tray Components of HPLC Column holder
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Several reservoirs (e.g., A1, A2 (usually aqueous ), B1,B2 (usually organic ), > 500mL) Precautions: 1. Degassing : remove of dissolved gas Sparging: fine bubble of gas, vacuum pumping, distillation, heating Degasser 2. Dust removal : Interference with detection, column clogging, damage Pumping system Millipore filter under vacuum Solvent Inlet filter Mobile phase reservoirs
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Pump 1. Displacement pump (Screw-driven syringe pump): pulse free, small capacity (250 mL), no gradient elution 2. Reciprocating pump Most widely used. Small internal Volume (35 ~ 400 μL), high pressure (105 psi), gradient elution, constant flow. Need pulse damper Requirements: high P (up to 6000 psi), pulse-free, constant Flow (0.1 ~ 10 mL/min.), reproducibility (0.5%), resistant to corrosion Isocratic elution: constant composition Gradient elution: different solvent systems during elution, continuous change or step wise, solvent proportion valve
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Reciprocating pump Seal Piston Motor Head outlet check valve Inlet check valve
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This note was uploaded on 01/05/2012 for the course CHM 410 taught by Professor - during the Fall '11 term at University of Toronto- Toronto.

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L11 - Lecture 11 Liquid Chromatography Liquid...

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