High-performance liquid chromatography is a separation technique in which isocratic and gradient mode is used to separate the analytes. Isocratic and gradient elution both are a useful model of separation but the isocratic elution is widely used one. In which a single solvent/mobile phase of constant composition is used to separate the compounds.
Isocratic elution is usually helpful in the separation of molecules that differ greatly in their affinity for the stationary phase. As long as possible, use isocratic elution, go to gradient only when isocratic doesn’t work.
The advantages of isocratic elution in HPLC are given below.
- It is simple to use
- Does not required to column re-equilibrate with the solvent/mobile phase between consecutive injections
- It has a lower cost
- This is the most reliable method
- The isocratic method is transferable
- No particular HPLC pump necessary
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