Difference Between Stationary Phase and Mobile Phase

What is Stationary phase in HPLC?

  • Depending on the type of stationary phase used, separations are done using partition, adsorption, or ion-exchange processes.
  • Modified silica or polymeric beads are the most widely used stationary phases. Long-chain hydrocarbons are added to the beads to make them different.
  • Refer Chromatographic Columns for details on HPLC column.
  • The size of the beads is frequently mentioned in the monograph. Changes in packaging type and size are described in this chapter’s System Suitability.
  • The stationary phase may or may not have interactions with the sample’s components.
  • Stationary phase can be packed column or silicon plates used in Thin Layer Chromatography (TLC)

Examples: C8, C18 column, TLC Plate

Related: Difference Between C18 and C8 HPLC Column.

Difference Between Stationary Phase and Mobile Phase

What is the basic requirement of a guard column in HPLC column?

  • In LC procedures, a guard column may be used with the following requirements, unless otherwise indicated in the individual monograph:
  • (a) The length of the guard column must be NMT 15% of the analytical column’s length.
  • (b) The analytical column’s inner diameter (d) must be the same as or smaller than the analytical column’s inner diameter (d), and
  • (c) The packing material ideal should be the same as the analytical column (e.g., silica) and contain the same bonded phase (e.g., C18).

In any case, all system suitability requirements specified in the official procedure must be met with the guard column installed.

Related: Principle of HPLC

What is Mobile phase in HPLC?

  • Mobile phase in chromatography is a compound used to separate components in a mixture.
  • The mobile phase is a solvent or a mixture of solvents, as described in the individual monograph. The mobile phase is either a gas or a liquid.
  • The mobile phase may completely dissolves the sample (Analyte).

Example: 50:50 Water: ACN, 75:50 Buffer: Methanol

Related: Number of theoretical plates (N), Resolution (RS), Column Storage, Column Cleaning and Regeneration of HPLC column.

Related: Difference Between Isocratic and Gradient Elution in HPLC

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