Thin Layer Chromatography

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Experiment 6 A & B: Thin Layer Chromatography Date: 10-9-12 Purpose: (A) Use thin layer chromatography to separate mixture of compounds of Fluorene, Fluorenol, & Fluorenone, then determine compounds found in unknown sample. (B) Determine development solvent by experimentation. Try 3 solvents for separating a pair of compounds that differ slightly in polarity. Procedure Outline:Part A1.Drew pencil line across plate 1 cm from bottom2. Marked 5 1 cm intervals on line starting 0.6 cm from edge of plate3. Prepared 5 micropipettes to spot plate4. prepared chamber with methylene chloride as solvent using a beaker with ½ sheet of filter paper and covered with aluminum foil.5. Spotted plate left to right with fuorene, fluorenol, fluorenone, unknown 3D, and reference mixture containing all 3 compounds a. used different pipettes to spot plate b. drew up sample by capillary action c. applied light spot (about 2 mm) d. allowed to evaporate6. Developed TLC plate by placing plate in chamber not contacting filter paper.7. Removed plate from chamber when solvent front was 1 to 2 cm from top of plate.8. marked solvent front with pencil line & set on paper towel to dry.9. Marked position of compounds on plate using a UV lamp10.Calculated Rf values for each spot11. determined results were unsatisfactory to how experiment was supposed to turn out.Part B1. Ran compounds of vanillin and vanillyl alcohol on TLC plates2. Prepared 3 TLC plates 10cm X 3.3cm3. Marked each plate with line 1 cm from bottom then with intervals 1 cm apart on lines for compounds to be spotted.5. Prepared 3 development chambers each with a different solvent. Acetone, 50% toluene & 50% ethyl acetate, and Hexane6. Spotted each plate left to right using micropipette with vanillin, vanillyl alcohol, and reference containing both compounds.7. Placed each plate into a development

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