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    FS Extraktion Boden

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    ericconchamunoz
    This document provides instructions for extracting fatty acids from soil samples. The process involves adding buffers, solvents, and water to the soil sample to extract the fatty acids, then separating lipids using a silica column and eluting with different solvents to isolate neutral lipids, glycolipids and phospholipids. Finally, the fatty acids are converted to fatty acid methyl esters (FAMEs) using a methylation reaction to make them suitable for gas chromatography analysis.

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    FS Extraktion Boden

    Uploaded by

    ericconchamunoz
    6 views1 page
    This document provides instructions for extracting fatty acids from soil samples. The process involves adding buffers, solvents, and water to the soil sample to extract the fatty acids, then separating lipids using a silica column and eluting with different solvents to isolate neutral lipids, glycolipids and phospholipids. Finally, the fatty acids are converted to fatty acid methyl esters (FAMEs) using a methylation reaction to make them suitable for gas chromatography analysis.

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    This document provides instructions for extracting fatty acids from soil samples. The process involves adding buffers, solvents, and water to the soil sample to extract the fatty acids, then separating lipids using a silica column and eluting with different solvents to isolate neutral lipids, glycolipids and phospholipids. Finally, the fatty acids are converted to fatty acid methyl esters (FAMEs) using a methylation reaction to make them suitable for gas chromatography analysis.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    6 views1 page

    FS Extraktion Boden

    Uploaded by

    ericconchamunoz
    This document provides instructions for extracting fatty acids from soil samples. The process involves adding buffers, solvents, and water to the soil sample to extract the fatty acids, then separating lipids using a silica column and eluting with different solvents to isolate neutral lipids, glycolipids and phospholipids. Finally, the fatty acids are converted to fatty acid methyl esters (FAMEs) using a methylation reaction to make them suitable for gas chromatography analysis.

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    Download as DOCX, PDF, TXT or read online from Scribd
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    Extraction of Fatty Acids For approximately 2g soil ( use glassware with screw caps!) Add 2ml Buffer (0.

    05M K2HPO4), 5ml MeOH and 2.5ml CHCl3 2h shaking Add 2.5ml H2O and 2.5ml CHCl3 Store overnight in the fridge (phase-separation) Transfer the (lower) Chloroform phase to clean class vials (Pasteur-glass-pipette) Removal of the solvent in N2-gasstream Add 1ml CHCl3

    Separation of Lipids Prepare a Silica column: push a piece glass wool in the glass column and add some Silicagel (Unisil) Add 5ml Ammoniumacetate (0.02M in MeOH) Wash the column: first with 5ml Aceton and second with 5ml CHCl3 Add the sample Add 5ml CHCl3 -Elution of Neutrallipids Add 5ml Aceton- Elution of Glycolipids Add 3x 5ml MeOH Elution of Phospholipids Collect the fractions of interest Removal of the solvent in N2-gasstream

    Methylation to FAME (Fatty Acid Methyl Esters) Add 600l BF3 in MeOH Incubate for 15min at 95C Add 0.3ml H2O and 0.5ml Hexane 60sec. shaking Transfer the (upper) Hexane phase to clean GC vials Removal of the solvent in N2-gasstream Add 0.5ml Hexane

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