How to use Beer-Lampert law without knowing pathlength?

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SUMMARY

The discussion centers on the application of Beer's Law using ATR-IR spectroscopy without knowing the pathlength. The formula for penetration depth is provided, which is essential for calculating concentration when pathlength is unknown. Users suggest constructing a calibration curve from absorbance data to determine concentration, emphasizing the need for prior measurements of different concentrations. The importance of understanding the incident angle and refractive indices is also highlighted as critical for accurate calculations.

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  • Understanding of Beer's Law and its components (absorbance, pathlength, concentration).
  • Familiarity with ATR-IR spectroscopy and its operational principles.
  • Knowledge of how to construct calibration curves in analytical chemistry.
  • Basic understanding of refractive indices and their relevance in spectroscopy.
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  • Learn how to calculate penetration depth in ATR-IR spectroscopy.
  • Research methods for constructing calibration curves from FT-IR data.
  • Study the significance of incident angle in ATR-IR measurements.
  • Explore the refractive indices of common solvents used in IR spectroscopy.
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Chemists, analytical scientists, and laboratory technicians involved in spectroscopy and concentration analysis using ATR-IR techniques.

rawr18
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So I did an experiment today using ATR-IR spectroscopy, thus I didn't know what the exact pathlength was (I should have just used traditional FT-IR!). Anyhow, I know that Beer's Law states that:

A(Absorbance)= a(wavelength) * b (pathlength) * c (concentration)

I know absorbance and wavelength, and am trying to find the concentration of my sample. However, I have no idea what the pathlength was since I used ATR. I looked up a formula online for penetration depth (which may be interchangeable with pathlength, not really sure), and it read:

dp= λ / 2(π)(n1) sqrt(sin^2θ - (n2/n1)^2)

I'm just confused as how to find the incident angle, ATR crystal refractive index and sample refractive index?

Any help would be much appreciated! thanks.
 
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Without the path length the Beer-Lambert law is redundant. One way round your problem would be to construct a calibration curve for your substance vs absorbance and then simply read off your concentration at the appropriate absorption
 
Thanks Text, is there a way to create a calibration curve AFTER the analyses have been done? I already have my data in the form of FT-IR graphs, how can I create a calibration curve?
 
rawr18 said:
is there a way to create a calibration curve AFTER the analyses have been done

If you have not recorded signals for different concentrations - no. If you did - it is not different from calibration curve for any other type of measurement.
 
Thanks Text, is there a way to create a calibration curve AFTER the analyses have been done? I already have my data in the form of FT-IR graphs, how can I create a calibration curve?

Make solutions with a range of concentrations which include your estimated value and measure their absorbances. Then plot concentration vs absorbance.
 
Maybe you can use some absorption of your solvent for calibration?
 
rawr18 said:
I'm just confused as how to find the incident angle, ATR crystal refractive index and sample refractive index?


For the first two, RTFM of the instrument. For the latter one, if your substance is diluted, you can probably use the index of the pure solvent (in the IR, of course).
 
Here's a dopy question - how do you get to display the originally posted questions when answering?
 
Hang on! I think I may have it! You just click "quote" right?
 
  • #10
tex43 said:
Hang on! I think I may have it! You just click "quote" right?

Yes.
 

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