Micro-extraction of urine and/or stomach contents and analysis by TLC

1342 Words Dec 10th, 2013 6 Pages
Micro-extraction of urine and/or stomach contents and analysis by TLC


Thin Layer Chromatography (TLC) is one of the oldest chromatographic techniques used for the identification of compounds and for determination of the presence of trace impurities.

The aim of the experiment was to determine two unknown samples (X and Y) by comparing results to the known standard drugs, of which there was five, using a specific TLC method.

The same technique was used for the determination of the unknown; however, the solution used for each method was slightly different. Furthermore, the mobile phase in each method was of different concentration with the addition of ethylacetate in Solvent system A.

The distance moved by the spot
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The two TLC plates were taken and the samples were spaced and marked appropriately with a pencil on each plate from 1 to 5 with X on one plate and Y on the other. Solution number 1 was paracetamol, 2 – salicylic acid, 3 – quinine, 4 – phenytoin and 5 was atropine. A capillary tube was used to take up each standard solution and was then gently touched against the TLC plate in the appropriate place on the line. A different capillary tube was used each time. Solution X was placed at the end of the line on one TLC plate and on the other, solution Y was spotted.

The TLC place with solution X on was placed in Beaker A, which contained Solvent System A and the other TLC placed was placed in Beaker B, which contained Solvent System B. Both beakers were then covered with foil and allowed to develop for 15 minutes.

Both TLC plates were removed from the beakers and the solvent front was marked with a light pencil. A hairdryer was used to speed up the process of drying. The TLC plates were then placed under a UV lamp to detect any visible spots and with a pencil were circled. The color of the spots was also identified while under the UV lamp.

Results and Discussion

Unknown X
Development time – 15 minutes
Distance migrated my solvent front - 61mm
Distance moved by spot (mm)
RF= Distance moved by spot Distance migrated by solvent

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