How to use Beer-Lampert law without knowing pathlength?

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In summary, the conversation discusses the use of ATR-IR spectroscopy and the issue of not knowing the pathlength when using this method. The Beer-Lambert law is mentioned, along with a formula for calculating penetration depth. The conversation also touches on the possibility of creating a calibration curve after the analyses have been done and finding the incident angle, ATR crystal refractive index, and sample refractive index. Suggestions are made for creating a calibration curve and using the index of the pure solvent for the sample's refractive index. The conversation also briefly mentions how to display the originally posted questions when answering.
  • #1
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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  • #2
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
 
  • #3
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?
 
  • #4
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.
 
  • #5
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.
 
  • #6
Maybe you can use some absorption of your solvent for calibration?
 
  • #7
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).
 
  • #8
Here's a dopy question - how do you get to display the originally posted questions when answering?
 
  • #9
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.
 

1. Can I still use Beer-Lambert law without knowing the pathlength?

Yes, you can still use Beer-Lambert law without knowing the pathlength by using a spectrophotometer or colorimeter. These instruments measure the absorbance of a solution and use a pre-determined pathlength to calculate the concentration of the absorbing species.

2. How accurate is the Beer-Lambert law without knowing the pathlength?

The accuracy of using Beer-Lambert law without knowing the pathlength depends on the accuracy of the instrument used to measure absorbance. If the instrument is calibrated correctly and has a high precision, the results will be accurate.

3. Can I estimate the pathlength in Beer-Lambert law?

Yes, you can estimate the pathlength by using a cuvette with known dimensions and calculating the pathlength based on the volume of the solution and the dimensions of the cuvette. However, this estimation may not be as accurate as using a spectrophotometer or colorimeter.

4. Is the Beer-Lambert law applicable to all types of solutions?

The Beer-Lambert law is applicable to dilute solutions of non-interacting molecules. It may not be accurate for solutions with high concentrations or those that contain interacting molecules or particles.

5. Can I use Beer-Lambert law for mixtures of different absorbing species?

Yes, Beer-Lambert law can be used for mixtures of different absorbing species as long as each species has a distinct molar absorptivity and the concentrations are low enough to follow the linear relationship between absorbance and concentration.

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