Cyclohexane Chromatography Essay

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Recommended: Using chromatography for seperating photosynthetic pigments

Procedure:
Extracting the pigments- Combined five milliliters of spinach in ethanol with five milliliters of cyclohexane in a test tube. Inserted a test tube stopper and shook. Once settled, pipetted the top layer into a small glass container. Added another five milliliters of cyclohexane to the spinach and repeated.
Separating remnants of polar solvents- Added ten milliliters of water to the cyclohexane, carefully stirred and added a solution of brine when an emulsion appeared, then carefully stirred again. Pipetted the top layer into another glass container, added water again, stirred, and pipetted the top layer into an erlenmeyer flask. Used a spatula to scoop anhydrous sodium sulfate into the cyclohexane mixture until it no longer stuck together at the bottom of the flask. Set up and ran the vacuum filter.
Concentrating the sample- Once all of the cyclohexane was filtered into the round bottom flask, removed and connected it to the rotary evaporator and allowed evaporation until the flask fully dried. …show more content…

Prepared 20 milliliters of eluent two by combining 10 milliliters of eluent one with 10 milliliters of cyclohexane in a beaker. Prepared 20 milliliters of eluent three by combining 10 milliliters of cyclohexane with 7 milliliters of eluent one and 3 milliliters of acetone in a beaker. Drew the starting point on three TLC plates, then applied one spot of the concentrated sample on each of the three TLC plates. Ran each TLC plate with one of each eluent systems to determine the best eluent, marked the stopping point and eluent used on each plate. Used the UV light to decide which eluent yielded the most spots. Eluent three showed the most spots. Made approximately twenty milliliters of eluent three in a beaker, marked it, covered it with a watchglass and set it aside for

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