Lecture+5 - Experimental variables in HPLC Pumping systems...

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Experimental variables in HPLC Pumping systems – 1 atmosphere = 101.325 Kilopascal; 414 bar = 408.6 atm=6004.5 pound/inch 2 Mobile phase isocratic elution (constant composition) The analyte interacts gradient elution (variable composition) with both, the mobile Flow rates : 0.1 to 10 mL/min and the stationary phase Column Dimensions (5 to 25 cm) Particle size (3 to 5 μ m) Stationary phase (pellicular and porous particles) Silica particles coated with organic films Temperature from RT to ca. 150 o C Sample size Volumes < 500 μ L
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Liquid chromatography modes
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2 24 ex M ru H D π = D M : diffusion coefficient of the analyte in the mobile phase in cm 2 /s m μ m m m m Band broadening
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Reciprocating pump Solvent consumption
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Bonded-phase partition chromatography The equilibrium between the mobile phase and a stationary phase comprising of either a liquid adsorbed on a solid or an organic species bonded to a solid is described as ‘partition’. The predominant type of separation in HPLC today is based on partition using bonded stationary phases .
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Lecture+5 - Experimental variables in HPLC Pumping systems...

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