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Applying too much compound to the TLC plate makes it difficult for a good separation to occur. Instead, a large blot of solution tends to appear in the TLC. "It is extremely important that the spots be as small as possible and that they be applied using a 1% solution of the compounds being separated

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How too much sample applied affect the TLC separation?

Applying too much sample on a TLC plate can lead to overcrowding of the compounds, causing them to run close together or even overlap. This can result in poor separation and difficulty in distinguishing different compounds. It may also lead to band broadening and decreased resolution, affecting the accuracy of the analysis.


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