Parameters that Characterize
HPLC Analysis
O U T L I N E
2.1. Parameters Related to HPLC
Separation 53
General Comments 53
Flow Rate 54
Retention Time 54
Retention Volume 55
Migration Rate 56
Capacity Factor (Retention Factor) 57
Equilibrium Constant and Phase Ratio 58
General Equation of Solute Retention 59
Characteristics of Ideal Peak Shape
and Definition of Efficiency 60
Selectivity 64
Resolution 64
Peak Capacity 66
Statistical Moments for the Description
of Peak Characteristics 67
Description of Peak Characteristics
for Gradient Separations 68
Quantitation in HPLC 69
2.2. Experimental Peak Characteristics
in HPLC 71
Van Deemter Equation 71
Kinetic Plots 75
Peak Asymmetry 76
Application of van Deemter Equation 78
Peak Characterization Using Exponentially
Modified Gaussian Shape 79
Summary of Chromatographic Peak
Characteristics 81
2.1. PARAMETERS RELATED TO HPLC SEPARATION General Comments Every analytical technique is characterized by its specificity/selectivity, reproducibility and repeatability, accuracy, range of linearity between the quantity of analyte and the response of the analytical instrument, limit of detection (LOD), limit of quantitation (LOQ), recovery yield of the sample processing, robustness, ruggedness, and stability [1]. In order to achieve optimum method