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Great! The spots are placed at a reasonable height and well spaced out.
Great! The spots are placed at a reasonable height and well spaced out.
Preparation of TLC plate.
Alt text: TCL plate marked for spotting. Samples are added to their respective location with a small capillary tube. The TLC plate is removed when the solvent reaches approximately 80% of the plate height, and the solvent front is marked with a pencil.
Performing the TLC analysis
Performing the TLC analysis
Materials needed: TLC plate, pencil, spotter, UV lamp
Materials needed: TLC plate, pencil, spotter, UV lamp
Gently mark the starting positions of the analytes on the TLC plate using a pencil. Each spot should be about 1 cm away from the bottom and labeled with a number or a letter. The last lane should be the cospot lane. CAUTION: The silica layer on the TLC plate is fragile, so do not press the pencil too hard.
Use a spotter to apply small samples of analyte solutions to each desired position on the plate. Allow the solvent to evaporate. Visualize the spots under UV light and redo them if they are not visible. CAUTION: Do not look directly at the UV lamp lightbulb when it is on as it can damage your eyesight.
Place the TLC plate in the developing chamber so that the bottom edge is immersed in the developing solvent and the plate is propped against the side of the chamber. Ensure that the analyte spots are above the developing solvent level when the plate is added to the chamber.
When the developing solvent has reached >80% of the height of the TLC plate, remove the plate from the chamber. Mark the location of the solvent front immediately and then allow it to evaporate.
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