UV-Vis spectroscopy: Changes in refractive index

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Controlling for changes in refractive index when obtaining UV-Vis spectra of aqueous solutions in a double beam spectrometer is crucial for accurate absorbance measurements. The incident beam is typically perpendicular to the surface of the cuvette, which minimizes refraction but can still affect reflection. Changes in refractive index can alter reflection, impacting absorbance readings. Users are encouraged to perform their own reflectivity calculations using the Fresnel equation, considering the refractive indices of air, water, and the cuvette material (often quartz). This involves estimating how dissolved substances affect the refractive index and, consequently, the reflectivity. While the initial question may seem straightforward, understanding the underlying principles requires deeper engagement with calculations and concepts, rather than seeking simple yes or no answers. Ultimately, the effect of changes in refractive index on absorbance measurements is considered negligibly small.
Ella98
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I wanted to know how do you control for changes in refractive index when you get the UV-Vis spectrum of an aqueous solution in a double beam spectrometer? Is that something the equipment does automatically? Or something I have to do? Also, how do these changes in refractive index occur? Thanks for any help you can provide.
 
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How does the beam hit the surface of the liquid? At what angle?
 
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Borek said:
How does the beam hit the surface of the liquid? At what angle?
The beam is perpendicular to the surface of the cuvette holding the liquid. Thanks!
 
Ella98 said:
The beam is perpendicular to the surface of the cuvette holding the liquid. Thanks!
It is not going to refract if the incident beam is perpendicular to the surface. Changes in refractive index will change the reflection though.
R=\left |\frac{n_{1}-n_{2}}{n_{1}+n_{2}} \right |^{2}
 
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HAYAO said:
It is not going to refract if the incident beam is perpendicular to the surface. Changes in refractive index will change the reflection though.
R=\left |\frac{n_{1}-n_{2}}{n_{1}+n_{2}} \right |^{2}
Will changes in reflection affect my absorbance measurements? I just want to know if the changes in refractive index will affect my absorbance measurements. Thanks, Hayao.
 
It might be nice if you could attempt a reflectivity calculation on your own of the case where there is only water in the cuvette, and then think about how much that changes when there is a sample dissolved in it. Don't ask for "do it for me". Try your own attempt and we'll guide you.To start you off, look up refractive index for air, water, and cuvette (most likely made of quartz). And then put the numbers in the Frensel reflectivity equation above for both intersection, namely air-quartz and quartz-water. At this point, you will have to make an assumption based on these numbers whether you can neglect multiple reflection (reflection of a reflection of a reflection of a...) or not. Then calculate how much of the initial light passes through the cuvette with water. (Note: we have four intersection in total, i.e. two quartz walls.)

If you have done that, then make a reasonable estimate of how much the refractive index of the water changes when you have something dissolved in it. Contemplate how much that would affect the reflectivity.
 
HAYAO said:
It might be nice if you could attempt a reflectivity calculation on your own of the case where there is only water in the cuvette, and then think about how much that changes when there is a sample dissolved in it. Don't ask for "do it for me". Try your own attempt and we'll guide you.To start you off, look up refractive index for air, water, and cuvette (most likely made of quartz). And then put the numbers in the Frensel reflectivity equation above for both intersection, namely air-quartz and quartz-water. At this point, you will have to make an assumption based on these numbers whether you can neglect multiple reflection (reflection of a reflection of a reflection of a...) or not. Then calculate how much of the initial light passes through the cuvette with water. (Note: we have four intersection in total, i.e. two quartz walls.)

If you have done that, then make a reasonable estimate of how much the refractive index of the water changes when you have something dissolved in it. Contemplate how much that would affect the reflectivity.

I'm sorry I didn't mean with my reply to imply I wanted you to do any calculations for me. I just thought what I asked was a yes or no question. I didn't know calculations will be required to answer it. Thanks for the help! I really appreciate it.
 
Ella98 said:
I'm sorry I didn't mean with my reply to imply I wanted you to do any calculations for me. I just thought what I asked was a yes or no question. I didn't know calculations will be required to answer it. Thanks for the help! I really appreciate it.
And that's not what I meant, neither.

You can't understand something with simple yes or no. You understand it by really doing the calculations on your own, even if you intuitively understand why. I wanted you to really understand it. What if you had to teach someone how to use UV-Vis absorption spectrometer and you were asked the same question? Yes or no isn't always the answer that they are looking for.The answer is it's negligibly small enough, btw.
 
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