- 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.
- 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.
- 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.
- 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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