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Separation of Compounds Using Column Chromatography




  1. Preparation of the Column:


    • Place the column in a ring stand in a vertical position.
    • A plug of glass wool is pushed down to the bottom of the column.
    • Prepare slurry of silica gel with a suitable solvent & pour gently into the column.
    • Open the stop cock & allow some solvent to drain out. The layer of solvent should always cover the adsorbent; otherwise cracks will develop in the column.

  3. Adding the Sample to the Column:


    • Dissolve the sample mixture in a minimum amount of solvent (petroleum ether).
    • Remove the solvent by placing the mixture in a rotary evaporator at a low temperature.
    • Place the dry powder on a piece of weighing paper and transfer it to the top of the column through the funnel.

  5. Developing the Chromatogram:


    • Attach a dropping funnel filled with petroleum ether on to the column.
    • Add petroleum ether continuously from the funnel to the top of the column.
    • Open the stopcock carefully.
    • The components of the mixture run down the column forming two separate yellow bands.

  7. Recovering the Constituents:


    • Continue running the petroleum ether till both the bands are eluted out separately.
    • Collect the constituents in two different R.B flasks. (Ortho nitrophenol is obtained first, followed by para nitro phenol.).
    • Evaporate the solvent by placing the mixture in a rotary evaporator.

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