 TLC

+ ADSORPTION chromatography

Purpose: separate non-volatile mixture, assess the purity of substances, confirm the identity of the
compound

Stationary phase: sheet of inert substrate: glass, plastic, or aluminium foil, which is coated with a thin
layer of adsorbent material, usually silica gel, aluminium oxide (alumina), or cellulose. (polar)

Mobile phase: solvent drawn up by capillary action (non-polar)

Visualize by: UV light onto the sheet treated with phosphor or chemical processes producing color
(developing agent)

total distance traveled by the sample

The results: retardation factor = 0 < <1
the distance traveled by the solvent
Strong interaction between analyte and stationary phase: 0

Strong interaction between solvent and stationary phase: 1

Separate based on difference of: + molecular size + polarity

The distance of the analyte depends on: solubility in the solvent + how well it adheres (dính) on the
plate +polarity

 The best separation occurs when the difference between the distances travelled by the compounds is
greatest.

 Isocratic elution:

Characteristics:

Solvent composition is constant

No equilibration necessary

Long analysis time

 LIQUID CHROMATOGRAPHY:

4 types:

+ Adsorption

+ Partition

+ Ion exchange: electrostatic attraction

+ Size-exclusion

In reversed phase chromatography the mobile phase is more polar than the stationary phase. Mobile
phase in reversed phase chromatography usually consist of water or buffer and an organic
modifier. The retention time is based on the hydrophobicity of an analyte molecule, which can be
expressed as log(P). The retention time is (amongst others) influenced by the solvent strength of the
organic modifier and the temperature.

Column chromatography:

Principle: ADSORPTION

Separation based on the polarity difference of the solute

Adsorb to mobile phase -> move a lot

Adsorb to stationary phase -> move a little bit

Elution order: increasing polarity and increasing distribution ratio

Elution power (eluent strength): its overall polarity + polarity of the stationary phase + nature of the
sample components

+Stat: solid

+ Mobile: liquid

Strength of adsorption:

Alkane > ester > ketone >aldehyde > phenol

 Planar chromatography: mobile phase moves through the stationary phase by the influence of
gravity or capillary action
 Reversed-phase: stationary phase is non-polar

Eluant: column

Eluate: mobile phase leaving the column

Eluent: solvent carrying the analytes

 GC: mobile phase: volatility

Stationary phase: boiling point