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Experiment 5: Separation of Unknown by Column Chromatography | Julianny Perez | Michelle Linder | 10/22/2013

Abstract The purpose of this lab is to introduce students to different methods of Chromatography which is widely used in the chemistry laboratory as a method of purification and analysis. Chromatography is the most important method of purification and it will be demonstrated through Column Chromatography where the stationary phase consist of a solid absorbent and the mobile phase is a liquid. Three eluents will be collected and tested using Thin Layer Chromatography to test its purity. Introduction Column Chromatography is used to isolate certain compounds from an entire mixture. Usually, the mixture is place on top of the solid absorbent and then a solvent is applied to run the mixture through the solid absorbent. Compounds that are in the mixture will have different interactions with the liquid solvent and the solid absorbent and as the mobile phase runs down the solid, separation will be achieved through varying degrees [3]. This experiment will demonstrate how a mixture containing Benzoin and Benzil can be separated using column chromatography. In this method of chromatography, a solid (in this case dry alumina) is added to a Pasteur pipet to act as a filter when running through three types of liquid Eluent with varying degrees of solubility. The three eluents collected at the end of the column chromatography will then be analyzed by Thin-Layer Chromatography to test its purity Chemical Reaction

Safety This lab used all safety precautions. Full sleeved shirts, pants, and shoes were worn in the lab at all times including splash-proof goggles. Gloves were worn in the experiment and all reactions containing alumina were handled under the hood to avoid inhalation. Benzil, Benzoin and Alumina were used in this lab which all were hazardous in case of skin contact, eye contact, and inhalation so all appropriate measures were taken for safety.

Experimental

Experiment 5: Separation of Unknown by Column Chromatography | Julianny Perez | Michelle Linder | 10/22/2013

The procedure for this experiment was found on page 7 and 8 of the lab handout [3]. The only changes to this experiment occurred in step 1 the Experimental part B on page 9 where we were instructed to place a 2.5 x 6 cm piece of paper towel in a 150ml beaker. This step is used to saturate the atmosphere with the 8:2 Hexanes: Diethyl Ether. Instead, we placed a watch glass on top of the beaker and stirred the mixture around.

Results When the mixture containing Benzil and Benzoin were added to the column chromatography apparatus, a noticeable yellow pigment was seen on the top. Once the first eluent was added, this yellow pigment was seen gradually traveling down the alumina until it all dripped down the flask. Once the yellow pigment was predominantly gone, 10 ml of the second eluent was added/collected and a final third eluent of 10 ml was added/collected. The amount of product recovered was then evaporated and weighed. From the final product weighed, we took dabs of each eluent and placed it on three different spots in a labeled TLC plate along with pure known Benzil and Benzoin on the 4th and 5th position respectively to compare the purity on each eluent, Part A Data Compound Chromatography mixture 1:1:3 Benzoin Benzil Alumina 9:1 hexane ether 8:2 hexane ether 1:1 hexane acetone Part B Data Compound Solvent Front in cm= 4.0 Spot # Distance UV Traveled S/W/I* (cm) 1 1.7 S Iodine (+)(-) XXX Rf Molecular Weight XXX 212.24 g/mol 210.235 g/mol XXX XXX XXX XXX Total Amounts of Material Used Recovered .15 g XXX XXX XXX 1.55 g 7.5 ml 10 ml 10 ml

X 1.022 .363 .066

Fraction 1 (9:1 Hex Ether) Fraction 2 (8:2 Hex Ether) Fraction 3 (1:1 Acetone:

.425

1.6/1

XXX

.4/.25

.5/1.6

XXX

.125/.4

Experiment 5: Separation of Unknown by Column Chromatography | Julianny Perez | Michelle Linder | 10/22/2013

Hex) Benzil 4 .5 S XXX Benzoin 5 1.7 I XXX S= strong absorption dark , W= weak absorption, I=invisible on UV

.425 .125

*Not to scale

Discussion After application of the TLC plate in the solvent, the solvent front was visible on the 4cm mark. Under the UV light spectrum, Eluent 1 which contained mostly the yellow pigment was only seen as one dot 1.7 cm above its original line. This corresponds to the same distance traveled by the compound on spot # 5 which was pure known Benzoin which means the column chromatography was effective in separating Benzoin from the mixture with 9:1 hex ether. The second eluent had two dots: one 1.6 cm above its origin and another 1 cm above its origin. Eluent 2 contained a mixture of Benzoin and an unknown that most likely appeared due to contamination of our beaker (because of not cleaning it properly before adding eluent 2). Eluent 3 contained a mixture of mostly Benzil since it corresponded to the same dot formed in position 4 of the TLC plate and was much darker with a little bit of Benzoin which had a light color. It wouldve been better for future experiments to get a more precise ruler as we used the end of our lab notebooks, which couldve had better results.

Conclusion The eluent 1 we collected in part A was Benzoin. The eluent 2 contained Benzoin and a contaminant. Eluent 3 contained both Benzoin and Benzil, but had Benzil in a higher concentration. References 1. Horowitz, G. J Chem. Educ. 2000, 77, 263. 2. Univ. of Colorado, Boulder, Chemistry & Biochem Department website http://orgchem.colorado.edu/hndbksupport/colchrom/colchrom.html 3. Prof. R. Ronald (Chromatography lecture notes) Department of Chemistry Washington State, University Pullman, WA