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
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