Extraction of Spinach- Lab 3

Extraction of Spinach- Lab 3 - Organic Chemistry 235 Lab...

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Organic Chemistry 235 Lab Lab 3 Extraction of Spinach Pre-Lab. Purpose of Experiment. To isolate different compounds present in spinach by using column chromatography. Precedure. 1) The fresh spinach leaves can be prepared by for the whole class or individually. 2) Pour organic portion in to a beaker. This should be very dark green color, if not then add acetone to portion. 3) Pour 3-4 ml into each students centrifuge tube.Centrifuge for 1 minute.Then transfer organic layer into test tube. 4) Add anhydrous sodium sulfate to dry the solution. 5) Add minimal amount of hexanes and transfer to test tube marked E for extract. 6) Dry pack a microscale column in a 5” Pasteur pipet with alumina. 7) Use hexanes as solvent to moisten the column( do not let column dry) 8) Put about 5 ml of concentrated green solution on the column. 9) Continue collecting the solvent eluting from the column (test tube 1)until the orange band(carotene) is close to the bottom. 10) Collect orange band and put in test tube #2 11) Add 75/25 hexanes/acetone solution.Use 4ml of mixture and collect the eluent in test tube #3 12) Once green band has reached bottom of test tube, switch to test tube #4.Six ml of acetone should be sufficient to elute all organic material off the column. 13) Prepare TLC analysis. 75/25 hexanes/acetone mixture in chamber. Spot four different solutions on the TLC plate using capillary about 1.5-2cm from bottom of plate. 14) Run the TLC plate.Remove plate from chamber when solvent is about 1-2 cm from top.
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This note was uploaded on 10/19/2008 for the course CHM 235 LAB taught by Professor Kolchens during the Spring '08 term at Pima CC.

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Extraction of Spinach- Lab 3 - Organic Chemistry 235 Lab...

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