231L+Dyes+TEMPLATE 2023-04-18 20_33_05
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Iowa State University *
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Feb 20, 2024
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NEW TEMPLATE: 231L Synthetic Organic Dyes For this experiment you will by synthesizing an organic dye. You will use absorption spectroscopy, fluorescence characterization, and TLC to identify your dye. Pre-lab Questions Procedure and Safety 1.
(10 pts) Write a bullet list outline for the procedure. 2.
(5 pts) Write a brief summary of the key safety concerns specific for this experiment. Data and Observations (Only record for dye that you synthesized.) 1.
(35 pts) Record what you are instructed to record in the procedure. In addition, while you are performing the synthesis of the dye that you chose, record any issues you had or what you could have done differently in order to get better results. There were no isssues in this lab, however I did get the wrong solution for the absorbance test. That issue was caught before getting the data though. REQUIRED DATA AND OBSERVATIONS FOR BOTH DYES (Record data for the dye that you synthesized. Obtain data for the other dye from a classmate.) 2.
(5 pts) Insert a photo of dye 1 after rotovaping (to show color of compound).
3.
(5 pts) Insert a photo of dye 2 after rotovaping (to show color of compound). (the other team did not take a picture of this) 4.
(18 pts) Insert three photos showing absorption spectrum for dye 1, fluorescein, and 9-
fluorenone. Indicate the wavelenth where maximum absorption occurred in each spectrum. Be sure to label each spectrum.
5.
(18 pts) Insert three photos showing absorption spectrum for dye 2, eosin Y, and rhodamine B. Indicate the wavelenth where maximum absorption occurred in each spectrum. Be sure to label each spectrum.
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6.
(18 pts) Insert three photos showing fluorescence for dye 1, fluorescein, and 9-fluorenone. Be sure to label which photo is which compound.
7.
(18 pts) Insert three photos showing fluorescence for dye 2, eosin Y, and rhodamine B. Be sure to label which photo is which compound. (other team didn’t provide 3 photos)
8.
(18 pts) Insert a photo of the TLC plate showing the synthesized dye 1, fluorescein, and 9-
fluorenone. The solvent front should be marked and circles drawn around any spots visualized using UV light. Plates should be clearly labeled with developing solvent used and what was spotted. Photo should include a ruler for calculating R
f
values.
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9.
(18 pts) Insert a photo of the TLC plate showing the synthesized dye 2, eosin Y, and Rhodamine B. The solvent front should be marked and circles drawn around any spots visualized using UV light. Plates should be clearly labeled with developing solvent used and what was spotted. Photo should include a ruler for calculating R
f
values.
Analysis and Application Questions Analysis 1.
(5 pts) Calculate the R
f
values for the spots on the TLC plate comparing dye 1 to fluorescein and 9-fluorenone. Show your work. 2.
(5 pts) Calculate the R
f
values for the spots on the TLC plate comparing dye 2 to eosin Y and rhodamine B. Show your work. 3.
(30 pts, 10 pts for specifically explaining how each type of data supports claim) What is the identity of dye 1? Use absorption spectrum including wavelength of maximum absorbance, fluorescence, and TLC, as evidence for your choice. Explain your choice. Remember that you need to be specific when using data as evidence
of your claim for the identity of dye 1. 4.
(30 pts, 10 pts for specifically explaining how each type of data supports claim) What is the identity of dye 2? Use absorption spectrum including wavelength of maximum absorbance, fluorescence, and TLC, as evidence for your choice. Explain your choice. Remember that you need to be specific when using data as evidence
of your claim for the identity of dye 2. 5.
(10 points) Suggest one way you could have improved your results based on the issues you recorded while performing the experiment.
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Application Your class has synthesized two dyes of drastically different colors. Understanding the relationship between compound structure, absorbed color, and observed color is fascinating and hugely important in industries ranging from fashion to medical imaging. For these application questions we’ll be focusing on the relationship between absorbed color and observed color. 6.
(10 pts) Fill in the lambda max for each compound in the table below that you observed while doing the experiment. Compound Bond-line Structure
max (nm)
Eosin Y Fill in
Rhodamine B Fill in 7.
(20 pts) What color does each of these compounds absorb based on their lambda max? Which compound absorbs at the lower energy wavelength? (Use this cool website to find this out: https://academo.org/demos/wavelength-to-colour-relationship/) 8.
(10 pts) Use the color wheel below to answer this question. Based on lambda max for rhodamine B and eosin Y, which of the two should have a slightly more red-purple color? Explain.
An interesting thing to note is that the chromophore part (conjugated part) of rhodamine B and eosin Y are almost identical, but they absorb at different wavelengths. This is because the attached groups, which are different for the two compounds, act as auxochromes. Auxochromes cause the absorbed light to shift to a more visible, lower energy wavelength. 9.
(12 pts) For these next questions, we’ll focus on two compounds with the same chromophore but different auxochromes found in nature. One is beta carotene and the other is lycopene. Beta carotene gives the orange color to carrots and lycopene gives the red color to tomatoes. The two compounds are shown below labeled only with their lambda max. Based on lambda max, identify which compound is beta carotene and which is lycopene. Explain your choice. View this question as a challenge to see if you understand the relationship between absorbed and observed color. (i.e. don’t google the answer until you have answered the question yourself!)
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