Chromatography | Interactive Continuous Simulation | Paper Chromatography
Chromatography: Principles, Methods
Separation technique — stationary phase, mobile phase, retention factor, types of chromatography
🧪 Real-time Paper Chromatography
Solvent rises continuously. Components separate by different Rf values. Simulation loops automatically.
Red / Fast Yellow Blue / Slow
Solvent front rising — components separating by Rf
Water (polar)
Ethanol (moderate)
Hexane (non-polar)
Dye mixture
Marker inks
Chromatography is a powerful separation technique used to separate, identify, and purify components of a mixture. It exploits differences in how substances partition between a stationary phase (solid or liquid) and a mobile phase (liquid or gas). Components migrate at different rates, allowing separation. It is fundamental in chemistry, biology, forensics, environmental science, and pharmaceutical analysis.
Principle of Chromatography
The mixture is dissolved in the mobile phase, which carries it through the stationary phase. Each component has a different affinity for the two phases → different migration speeds → separation. The retention factor (Rf) is a key parameter:
Rf = distance travelled by compound / distance travelled by solvent front
Rf is characteristic for a given compound under fixed conditions. It helps in identification.
Types of Chromatography
📄 Paper Chromatography
Stationary phase: paper (cellulose). Mobile phase: solvent. Used for separating dyes, amino acids, simple mixtures.
🧪 Thin Layer Chromatography (TLC)
Stationary phase: silica gel or alumina coated on glass plate. Fast, high resolution, widely used in organic chemistry.
🔬 Column Chromatography
Stationary phase packed in a vertical column. Mobile phase flows through, fractions collected. Used for preparative separations.
💨 Gas Chromatography (GC)
Mobile phase: inert gas (He, N₂). Stationary phase: liquid coated on solid support. For volatile compounds.
💧 High Performance Liquid Chromatography (HPLC)
High pressure forces mobile phase through column. High resolution, quantitative analysis.
🧬 Ion Exchange Chromatography
Separates ions and polar molecules based on charge. Used in protein purification, water softening.
Retention Factor (Rf) and Identification
After development, the distances are measured. Rf values between 0 and 1 indicate separation. Unknown compounds can be identified by comparing Rf with standards.
Example: Rf (blue dye) = 3.2 cm / 6.0 cm = 0.53
Applications of Chromatography
Pharmaceuticals: Purity testing, drug analysis, metabolite detection.
Forensics: Analysis of ink, dyes, drugs, and toxicological samples.
Environmental: Monitoring pesticides, pollutants, water contaminants.
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