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andrade5 class high score - EXPERIMENT 5: GEL PERMEATION...

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EXPERIMENT 5: GEL PERMEATION CHROMATOGRAPHY (INFORMAL) Data: ___20__ / 20 pts • Table of elution volumes obtained for pepsin, hemoglobin, cytochrome c, and myoglobin. (10 points) • Table containing experimental molecular weight and literature molecular weight for myoglobin. (5 points) Note: The literature value must come from a primary literature source. • Table of absorbance values for pepsin. (5 points) Sample Calculations: __6___ / 6 pts Calculation of molecular weight for myoglobin. Results: __22___ / 24 pts Plot of Log Molecular Weight vs. Retention Volume (V r ). (18 points) Determination of the molecular weight of myoglobin. (6 points) Questions: ___27__ / 30 pts Question 1: 6 points Question 2: 6 points Question 3: 6 points Question 4: 6 points Question 5: 6 points Conclusion: __10___ / 10 pts Style (organization, spelling, neatness, grammar, etc…) __10___ / 10 pts Other Deductions (i.e., Late Lab Report, incorrect formatting, etc…) - _____ Score __95___ / 100 pts
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The Determination of the Molecular Weight of Myoglobin by Gel Permeation Chromatography Francisco Andrade Jessica Booe 5/27/10 Wesley Storm CHEM 241L 402, Morehead Labs Room 301 Pledge
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The goal of this experiment was to determine the molecular weight of myoglobin using gel permeation chromatography. Gel permeation chromatography can be used to estimate the molecular weight of unknown substances by using a series of standards of known molecular weight and measuring their retention volumes. A calibration curve constructed by plotting the logarithm of the molecular weight for each of the standards vs. the retention volume measured by each standard can be used to estimate the molecular weight of an unknown compound as long as it falls within the linear region of the curve 1 . The first calibration standard used was pepsin, and because it does not absorb in the visible region, the UV absorbance was measured to find the most intense fraction, as three amino acids in pepsin absorb in the UV region at 280.0 nm 1 . The results of measuring the absorbance, using a spectrophotometer, of each fraction as it eluted from the column are displayed below. The most intense fraction was the fraction with the greatest absorbance value, which was fraction 6 at an absorbance value of 0.213 au. Fraction
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This document was uploaded on 11/04/2011 for the course CHEM 241 at UNC.

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andrade5 class high score - EXPERIMENT 5: GEL PERMEATION...

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