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High Performance Liquid Chromatography (HPLC)

HPLC Instrumentation Elution Gradient elution in Liquid Chromatography is analogous to temperature programming in gas Chromatography . Increased eluent strength is required to elute more strongly retained solutes. RP-HPLC (increase organic, make more non-polar). NP-HPLC (increase solvent to make more polar) Gradient (gradual change) of eluent strength is used for many separations. Weakly retained solutes are eluted with solvent of low eluent strength. Second solvent is mixed with the first to elute the more strongly retained solutes. Gradual increase in eluent strength. Partition Chromatography Normal-phase Chromatography uses a polar stationary phase and a less polar solvent.

Electrochemical detection (ECD) for HPLC is an extremely selective and sensitive detection technique that is applied in a number of analyses such as the neurotransmitters dopamine, serotonin and noradrenalin. ... Particle Size Effect of Stationary Phase Smaller particles, more efficient packing, lower A term. Stronger solvent, greater affinity ...

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