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Mobile Phase

Definition

The solvent or mixture that flows over the stationary phase and carries analytes along. Polarity, viscosity and volatility set the separation; switching from chloroform to ethyl acetate on the same TLC plate can move a spot from Rf 0.2 to Rf 0.8.

Role
Carries the analyte through the stationary phase during separation
In liquid chromatography (TLC, HPLC, HPTLC)
Solvent or solvent mixture
In gas chromatography (GC/GLC)
Inert carrier gas (helium, hydrogen, or nitrogen)
Key factors affecting separation
Polarity, viscosity, and volatility of the mobile phase

Common questions

What is the mobile phase in chromatography?+

The mobile phase is the fluid (solvent or gas) that moves through the chromatography system and carries your analyte along with it. In liquid techniques like TLC and HPLC, it is a solvent mixture. In gas chromatography, it is an inert carrier gas like helium or hydrogen.

How does changing the mobile phase affect separation?+

Changing the solvent mixture properties like polarity, viscosity, and volatility directly changes how far analytes travel. For example, switching from chloroform to ethyl acetate on the same TLC plate can move a spot's Rf value from 0.2 all the way to 0.8.

Why use different mobile phases for different chromatography methods?+

Different methods require different carriers. Liquid chromatography needs solvents that dissolve your sample and flow through the stationary phase. Gas chromatography needs inert gases that won't react with the sample, which is why helium, hydrogen, and nitrogen are all used as carrier gases.

Related terms

Stationary Phase
The immobile material the mixture flows past. In paper it is water inside cellulose fibres; in classical column work and TLC it...
Resolution (Rs)
Rs = 2(tR2 − tR1) / (W1 + W2). Above 1.5 means baseline separation; below 1.0 means peaks overlap and quantitation is...
Rf Value
The retention factor, defined as the distance the spot travels divided by the distance the solvent front travels on the same plate....
ECD
Electron capture detector. A Ni-63 radioactive source produces a baseline electron current that is reduced when electron-affinic species (halogens, nitro groups, peroxides)...
FID
Flame ionisation detector. A hydrogen-air flame ionises C-H bond containing analytes and a collector electrode reads the ion current. Near-universal for organic...
Isocratic vs Gradient Elution
Isocratic means the mobile-phase composition stays constant through the run; simple, reproducible, but limited resolving power for wide-polarity mixtures. Gradient means the...
Limit of Detection (LOD)
The lowest concentration of analyte that produces a signal reliably distinguishable from the instrument noise, conventionally three times the standard deviation of...
MSD
Mass Selective Detector. GC-MS or LC-MS detector that identifies analytes by mass spectrum. The confirmation standard for drug and toxicology casework.
Partition vs Adsorption
Partition chromatography separates by differential solubility between two liquid phases (GLC, reversed-phase HPLC). Adsorption chromatography separates by surface binding to a solid...
Plate Theory
Model that treats a column as a series of theoretical plates. The Height Equivalent to a Theoretical Plate (HETP) measures column efficiency;...
Retention Time (tR)
Time from sample injection to peak maximum at the detector in HPLC or GC. Characteristic of an analyte under a defined column...
Rf (Retention Factor)
Distance travelled by the analyte divided by the distance travelled by the solvent front, on a TLC or HPTLC plate. A unit-less...

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