User:Maria Briscione/Notebook/Chem 571/2011/10/04

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Objective

Determine a concentration of MBP that yields an accurate UV-Vis Spectrum.

Description

Estimated concentrations of MBP at 0.2, 2.0, 10.0, 100.0, and 1000.0 µg/mL were analyzed in the UV-Vis. Based on the resulting spectrum the estimated 100 µg/mL dilution was used to determine the molar absorptivity. The accurate concentration of the estimated 100 µg/mL solution was initially determined. The known value of the true concentration of the "2 g/mL" solution was determined to be 2.66055 µg/mL by using the equation of the line of the protein + Bradford Reagent Standards ( y = 0.218x). The absorbance of the protein ("2 µg/mL") in water was the y value ( 0.116) and an x value (concentration) was determined to be 2.66055 µg/mL.

  • 0.2 µg/mL -> .053211 µg/mL
  • 2 µg/mL -> .53211 µg/mL
  • 10 µg/mL -> 2.665 µg/mL
  • 100 µg/mL -> 26.6 µg/mL
  • 1000 µg/mL -> 266.055 µg/mL


A graph of the molar absorptivity was then constructed using the absorbance of the 2.66055 µg/mL solution's concentration and absorbance. A peak appeared at 280 nm.

Data

Image:oct4_graphs.jpg

Notes

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