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High Performance Liquid Photography - Coggle Diagram
High Performance Liquid Photography
Column-based chromatographic technique
Phase of HPLC
Normal Phase
Consist of polar
SP
(such as silica modified with polar group)
and non-polar
MP
(such as non-polar organic solvent)
Mainly used for high polarity species and some ionic species
Separation occurs with increasing polarity
Reverse Phase
Consist of non-polar
SP
(such as silica modified with non-polar group)
and polar
MP
(such as polar solvent)
useful for separating compounds of low to medium polarity
Separation occurs with decreasing polarity
Types of Detectors:
UV Detector
Differential refractor
Fluorescence Detector
Infrared Detector
Conductivity Detector
Electrochemical Detector
Diode Array Detector
Mass Spectroscopy
General System
Liquid MP is pumped under high pressure (MP is a solvent mixture which will affect the separations obtained)
Short column packed with finely divided solid SP
Stationary Phase
Invariably bonded shape that constraint in mixture
Definition: refers to solid support contained within column
Column
Guard- protect and increase lifetime of column
Capillary-allow user to work with nL samples
Fast-designed to minimise time of chromatographic analysis
Preparatory-used to prepare bulk samples
microbore-used for analytical volumes
Examples
Octadecylsilyl (reverse phase)
Octylsilyl (reverse phase)
Phenyloxylsilyl (reverse phase)
Cyanopropylsilyl (normal phase)
Mobile Phase
Depends on SP and type of system (normal/reverse)
definition: solvent being continuously applied to column
Examples: THF, Acetonitrile, aqueous methanol, organic solvents, aqueous CH3CN
Types of system
Pumping
Sample Injection system
Polylyptic (several technique using some coumn)
Gradient (elution using 2 or more solvent)
Isociatic (elution containing 1 solvent)
APPLICATION
Forensic science- analysis of alkaloids
Purification- separating compounds from another
Chemical Separation- chiral separations
Pharmaceuticals- using diode arrays