Solving the Beer-Lambert Equation for Lactate Solution

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Discussion Overview

The discussion revolves around the application of the Beer-Lambert equation to determine the optical density of a lactate solution. Participants are exploring the calculations involved in using the equation, particularly in the context of a homework assignment.

Discussion Character

  • Homework-related
  • Mathematical reasoning
  • Technical explanation

Main Points Raised

  • One participant presents their calculations using the Beer-Lambert equation, noting a discrepancy between their results and an example graph.
  • Another participant points out that Beer's law is typically accurate only for absorption values between 0.1 and 1.0, suggesting that measurement difficulties may arise outside this range.
  • A participant highlights that the example graph shows a maximum absorption of 0.4 at 340nm, and they propose an alternative formula that seems to yield a closer approximation, although they express confusion about its reasoning.
  • One participant suggests that the slope of the best fit line to the graph could provide the value of ε.

Areas of Agreement / Disagreement

Participants express varying views on the accuracy of the Beer-Lambert law under different conditions, and there is no consensus on the correct approach to reconcile the discrepancies in the calculations presented.

Contextual Notes

Participants note potential limitations related to the accuracy of measurements outside the recommended range for Beer's law and the reasoning behind alternative formulas proposed for calculating optical density.

nobahar
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Homework Statement


I need to use the Lambert-Beer equation to plot optical density of a lactate solution.

Homework Equations


[tex]A=\varepsilon\ell C[/tex]
(The path length is 1 cm)

The Attempt at a Solution


This is one of my four samples (if you can show me where I'm going wrong with this then I can apply the correction to the others):
2.22 [tex]\mu[/tex]mol in 1.5ml, therefore there is 0.296 [tex]\mu[/tex]mol in 0.2ml
This info seems to match to the example graph I have.
But using the formula above, where [tex]\varepsilon[/tex]=6.22 for micromol/ml, I get:
6.22 x (0.296/0.2) micromol/ml = 9.2056
This doesn't match up with the graph I have! Any help appreciated! Please!:redface:
 
Last edited:
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Beer's law is accurate only for values of A from about 0.1 to 1.0. Experimentally, it is difficult to accurately measure absorption values outside this range.
 
That's the main problem, the results from the example graph are at a maximum of 0.4 for the absorption at 340nm (y-axis) when plotted against amount in micromoles up to 0.4 on the x-axis.
I can get a fairly close approximation (the graph itself is supposed to be an estimate) if I use the following formula:
(amount in micromoles x amount in ml) x 6.22. But this equation makes no sense; or at least I can't see the reasoning behind it.
 
The slope of the best fit line to the graph should give you ε.
 

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