Solving the Beer-Lambert Equation for Lactate Solution

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AI Thread Summary
The discussion focuses on using the Beer-Lambert equation to calculate the optical density of a lactate solution, specifically addressing a discrepancy in results. The user attempts to apply the equation A=εℓC with a path length of 1 cm and a given molar absorptivity of 6.22, but their calculated value does not align with an example graph. They note that Beer’s law is most accurate for absorption values between 0.1 and 1.0, which complicates their results since the example graph's maximum absorption is 0.4. The user proposes an alternative formula that yields a closer approximation but expresses confusion over its validity. Clarification on the reasoning behind the alternative equation and the slope of the best fit line is sought for better understanding.
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Homework Statement


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

Homework Equations


A=\varepsilon\ell C
(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 \mumol in 1.5ml, therefore there is 0.296 \mumol in 0.2ml
This info seems to match to the example graph I have.
But using the formula above, where \varepsilon=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:
 
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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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