The column or stationary phase selection is the most significant advance in analytical method development. Without a column, which is stable and high performance, the development of a reproducible and rugged method is not possible. The selection of the column is done based on information about the nature and analysis of solutes.
Usually, a lengthy column gives improved separation because of higher theoretical plate numbers. A column with a 5-µm particle size provides good reproducibility, efficiency, and reliability.The column efficiency is reported as the number of theoretical plates. The efficiency of the column in HPLC is dependent on various factors; some factors that affect column efficiency in HPLC are given below.
- The activity of the adsorbent
- Column length
- The dimension of the column
- The flow rate of the system
- Packing of the column
- Particle diameter
- Particle size distribution
- Quality of solvents
- The temperature of the column