Practical real world uses for Colorimetry

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Discussion Overview

The discussion centers on the practical applications of colorimetry, particularly in various industries and research settings. Participants explore specific examples of colorimetric techniques and their relevance in real-world scenarios, including healthcare and laboratory research.

Discussion Character

  • Exploratory
  • Technical explanation
  • Conceptual clarification
  • Debate/contested

Main Points Raised

  • One participant highlights the use of colorimetry in hospitals to measure hemoglobin content in blood as a common health indicator.
  • Another participant mentions the application of colorimetry in research labs for quick tests of various elements before conducting certified analyses.
  • A specific example is provided regarding the use of dimethylglyoxime to check for traces of nickel during protein purification on nickel columns.
  • Further elaboration is requested on the protein purification process, including the use of histidine tags and nickel affinity columns.
  • A cautionary note is raised about the Lowry protein assay, emphasizing the importance of calibration and potential interferences in colorimetric methods.
  • Another participant references numerous adaptations of the phosphomolybdate method for determining inorganic phosphate, suggesting that protocols can be obtained from biochemistry departments.
  • One participant shares their personal use of colorimetry to measure formaldehyde emissions, offering to provide more details privately.

Areas of Agreement / Disagreement

Participants express various applications of colorimetry, but there is no consensus on the best practices or limitations of specific methods. The discussion includes both supportive examples and cautionary advice, indicating a range of perspectives on the topic.

Contextual Notes

Participants note the importance of calibration in colorimetric methods and the potential for misleading results if not properly standardized. There are also references to specific protocols and historical methods that may require further context for understanding.

Who May Find This Useful

This discussion may be useful for students and professionals in chemistry, biochemistry, and related fields interested in the practical applications and methodologies of colorimetry.

crybllrd
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I need to give a short presentation about Colorimetry in my chemistry class.
I plan on using the first five minutes giving an example procedure about determining the concentration using MicroLAB's spectrophotometer hardware in our lab.
I would like to use that last five minutes of my presentation talking about what industries use colorimetry, and perhaps use some real world uses that we could relate it to.
Any ideas?
 
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One that goes on in every single hospital all over the world is to measure hemoglobin content of blood as a health indicator. I'd guess it is the most common.
In many research labs (like mine) we use colorimetry as a quick test for different elements before we send out for expensive certified analysis.
When we make proteins and purify on nickel columns we often use the reagent dimethylglyoxime to check for traces of nickel.
 
Thanks a lot, that is exactly what I was looking for.
Do you mind elaborating on the "make proteins and purify on nickel columns we often use the reagent dimethylglyoxime to check for traces of nickel" part? I am trying to do some further research on these topics.
Thanks again.
 
A convenient way to make small amounts of mutated proteins is to edit in a "tag" with six histidines at the end. It has an affinity for nickel and there are columns e.g. "Histrap" with nickel bound to a chelator. You pour the bacterium lysate through the column and only your favourite protein sticks. You then elute with an imidazole buffer (competitive for nickel) and collect your protein. Occasionally some nickel ions tag along. They give a red color with dimethyl glyoxime.
 
Google 'Lowry protein assay'.

Very widely used there is also a caution, which is a matter of principle for your talk.

It is good enough for a lot of work for e.g. serum protein concentration and much else (v. widely used). OK when you are looking at relative values, e,g, variations of proein concentration. It is usually standardised, calibrated, using bovine serum albumin.

But if you were to use it to determine the concentration of a particular pure protein and the absolute concentration is important I know for a fact that wrong and misleading results can be obtained if you do not calibrate it using the same protein whose absolute concentration is known by some other method. Maybe this example to too advanced for your talk but in general since there can be interferences for any colorimetric method by other substances in mixtures, it should be standardised with solutions as far as possible resembling those you are going to be testing, I don't know if I have been clear.
 
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There are zillions of improvements, modifications, adaptations, tweaks, kits deriving from the phosphomolybdate method for determination of inorganic phosphate that originate from Fiske and SubbaRow [(1925) J. Biol. Chem. 66, 375] , anyone in your biochemistry department can give you a protocol e.g. from manufacturers' pamphlets and catalogs etc. Very widely used.a perspective
http://www.garfield.library.upenn.edu/classics1981/A1981LY47200001.pdf
 
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I use it to measure formaldehyde emissions. PM me if you'd like more details.
 

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