Lab Report Liquid Chromatograpy

Topics: Chromatography, Solubility, Chemical polarity Pages: 5 (1304 words) Published: April 10, 2015
Title: Liquid Chromatography
Author: Gloria Contreras
Lab Partner: Jose Montanez
Instructor: Teresa Potter
Date Work Performed: January 13, 2015
Date Submitted: January 20, 2015

Abstract:
In this lab, liquid chromatography is used to separate the Red 40 and Blue 1 dyes inside of grape flavored Kool-Aid. It was determined that the 5% isopropanol will remove the Red 40 dye from the stationary phase. The 28% isopropanol will remove the Blue 1 dye from the stationary phase. The more polar molecule is removed with the more polar mobile phase, the less polar molecule is removed with the less polar mobile phase. This is where the principle “like dissolves like,” plays a factor in the separation of the dyes. Both molecules are relatively nonpolar but the size difference is what causes Red 40 to be a more polar molecule than Blue 1 because it is smaller. Introduction:

In this lab, liquid chromatography is used to separate the dye colors of a Kool-Aid mixture. The mixture is injected into a chromatography column, which in this lab is a C18 Sep-Pac. This is where the mixture lands on a substrate. The stationary phase may be polar or nonpolar. When a solvent is passed through the stationary phase, it will carry out the components of the same polarity. This is where the principle ‘like dissolves like,” comes into play. This principle states that a polar substance will dissolve in a polar solution and vice versa, where a nonpolar substance will dissolve in a nonpolar solution. The speed that the substance comes out depends of the polarity of the substance and how well it dissolves in the solvent.

Liquid chromatography is an essential tool when trying to determine what substances are used in materials we come in contact with every day. For example, such tool can be used to determine the substances in grape Kool-Aid. Two known substances found in Kool-Aid is Red 40 dye and Blue 1 dye. These dyes have been linked to causing children to become hyperactive, and/or cause individuals to have an allergic reaction. This may cause someone to think twice before drinking grape Kool-Aid. Being able to separate these substances allows for individuals to determine if a material is safe for human consumption and handling. Materials:

C18 Sep-Pac Cartridge
Two 10 mL Syringe with Male Luer Tip
One 10 mL graduated cylinder
Two 25 mL graduated cylinders
70% Isopropanol (2-propanol)
Grape Kool-Aid Drink Mix (unsweetened)
Distilled Water
Four 50 mL Beakers
Three 100 mL Beakers

Procedure:
Step 1: Obtain goggles and prepared Kool-Aid mixture.
Step 2: Prepare solvent and pre-treat C18 Sep-Pac.
Step 3: Inject Kool-Aid into Sep-Pac. Collect and discard effluent. Step 4: Elute the components of the grape Kool-Aid.
Step 5: Record the volume of the eluate. Record the volume when the red dye first appears in the Sep-Pac and record again when the red dye is no longer present. Record the volume of the eluate when the blue dye is present and record again when the blue dye is no longer resent. Step 6: Repeat step 4-6 twice and record data.

Step 7: Prepare solvents.
Step 8: Repeat step 2 and then 3.
Step 9: Elute components of grape Kool-Aid and separate by gradient process. Step 10: Place beakers of eluate in hood and allow to evaporate. Record observations. Step 11: Discard of waste. Clean Sep-Pac.

Data Table:

Red

Blue

Trial
1
Trial
2
Trial
3
Trial
Avg
Trial
1
Trial
2
Trial
3
Trial
Avg
VR (start)
0.6
0.6
0.4
0.53
1
1
0.8
0.93
VR (end)
1.6
1.6
1.2
1.47
4
3.2
2.2
3.13

Red

Blue

Trial
1
Trial
2
Trial
3
Trial
Avg
Trial
1
Trial
2
Trial
3
Trial
Avg
W = VR (end) - VR (start)
1
1
0.8
0.93
3
2.2
1.4
2.2
VRavg = VR (start) + 0.5 W
1.1
1.1
0.8
1
2.5
2.1
1.5
2.03
L
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
r
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
VM (mobile phase volume)
0.49
0.49
0.49
0.49
0.49...
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