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Separation of Blood Serum Lipids by Thin-Layer Chromatography

This lab session from IUG in 2012 demonstrates the separation of blood serum lipids using thin-layer chromatography. The method is based on the varying migration rates of different lipid fractions in a solvent, with identification done through iodine vapor. Reagents and materials required include various solvents and chromatography plates. The procedure involves mixing blood serum with an ethanol/ether mixture.

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Separation of Blood Serum Lipids by Thin-Layer Chromatography

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  1. Lab session 5 Separation of blood serum lipids by thin-layer chromatograghy IUG, 2012 TMZ

  2. Principle • The method is based on different migration rates of blood serum lipid fractions: • Phospholipids • Free fatty acids • Cholesterol • Glycerides In a thin layer of adsorbent as an organic solvent moves through it.

  3. The rate of fractions separated is dependent upon their relative polarities. • A iodine vapor is then used to identify the separated fractions.

  4. Reagents & materials • Ethanol/diethyl ether mixture (3:1) • Chloroform • N-hexane/diethyl ether/acetic acid mixture (73:25:2) serves as a chromatography solvent • Metallic iodine • Thin lyer chromatography plates silufol UV254 • A chromatography chamber • A water bath • A development chamber • A chromatogram viewing cabinet • Blood serum sample

  5. Procedure 1. Mix 1 ml of blood serum with 10 ml of ethanol/ether mixture

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