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Spectrophotometric Analysis Of Dyes

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Introduction: The objective of this experiment was to use a spectrophotometer to calculate the absorbance wavelength for the commercial dyes that were given. Light is composed of tiny particles that are called photons, just like matter is composed of tiny particles called atoms. Using the spectrophotometer you can see that different dyes absorb at different wavelengths. With all of the experimentation done the concentration, absorbance and the max wavelengths should be found. Figure 1: Schematic of a single beam UV spectrophotometer Materials: There was an array of materials needed to complete this experiment. These included .20 milligrams of red dye #3,yellow dye #5, spectrophotometer, volumetric flask, cuvettes, beakers, distilled …show more content…

Adjusting the integration time, strobe lamp, and boxcar width. After all of this is done inserting the black cuvette is the next step and storing the spectrum. Insert a blank cuvette with distilled water and then store the reference cuvette again. Next is making the dye solution in order to find the max absorbance. Mixing .20 milligrams of the commercial dye (red dye #3) with 250 ml of distilled water to get the initial concentration, the goal is basically to get the absorbency level below 1. Putting the dye solution in the blank cuvette and observe the absorbance level that it dorms. Repeat the steps five times making different dilutions to maintain multiple absorbance values. The second week was making another commercial dye solution and also testing the food dye brought into class. Calibrate the spectrophotometer the same exact way so that it does not mess up the calculations gathered. This week using yellow dye #5 mix .20 milligrams of this will 250 ml of distilled water in a volumetric flask. Make five different solutions, distilling the water each time, of new yellow dye solution and place each of the solutions in a blank cuvette to find the max absorbency of each the solutions. Next during week two the absorbency had to be found from the food that was brought in. Using a blank cuvette place the commercial liquid and calculate the concentration and absorbency and which dyes would be in this liquid. Results: Maximum wavelength of Red Dye #3 Solution Absorbance Wavelength Max Concentration Solution 1 0.489 A.U. 526 nm 9.1 x 10-6 mol/L Solution 2 0.188 A.U. 525 nm 1.8 x 10-6 mol/L Solution 3 0.705 A.U. 525 nm 1.5 x 10-5mol/L Solution 4 0.682 A.U. 525 nm 1.2 x 10-5 mol/L Solution 5 0.527 A.U. 525 nm 1.2 x 10-5 mol/L Maximum wavelength of Yellow Dye

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Question 4: List the 3 errors; • Adding too many drops of NaOH at the same time would affect the results because we can’t determine the exact equivalent point when the color changed. The results won’t be accurate and that will affect all the data that are dependent on the amount of NaOH to titrate. • Other error could be the hardness to notice a color change; we always use a white paper under the flask to determine when the color changes right away. And if we don’t use the white paper it will be hard to determine the color change and the amount of NaOH that was used to titrate it. • Also other source of error could be by not rising the burette with NaOH before we fill up with it, or it maybe they were rinsing it with a lot of NaOH which could affect the data recording for NaOH amount of titration.

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That mixture was then filtered through a coffee filter. Nine test tubes were prepared in order to perform this dye coupled reaction. One contained 5.0ml of the potato and pH buffer mixture, 2.0 ml of hydrogen peroxide, and 1.0 of guaiacol to serve as a blank for the spectrophotometer. Four test tubes were filled with 2.0 ml of hydrogen peroxide and 1.0 ml of guaiacol, used for measurement by the spectrophotometer, each. The last four were filled with 4.0 ml of the potato and pH buffer mixture and 1.0 ml of peroxidase.

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The third peak consisted of all four colors and finally the large peak consists of green and blue. Orange dyes would probably have the more noticeable peaks at the shorter wavelengths because both red and yellow both have peaks at the shorter wavelengths. Purple dyes would most likely have one distinct peak for a short wavelength and then one distinct peak for the longer wavelength because red has peaks at short wavelengths and blue has peaks at long wavelengths. The data observed looks almost nothing like the D2L data.

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The cuvette was placed in the spectrophotometer with the arrows, on both the cuvette and the SpectroVis, facing the same side. After the recording, the cuvette was removed from the SpectroVis and the content was poured back into the original volumetric flask. The absorbance as well as the maximum wavelength of each solution was recorded in Table 3 and

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Allie Fullmer C127 Lab 1 October 2015 Spectroscopy of Food Dyes Summary: Dyes are added to colorless food; there are nine food dyes certified for food use in the United States. The Food and Drug Administration demands that all food dyes undergo an approval process. In this experiment a spectrometer was used to measure the absorbance spectrum of different food dye solutions. The absorbance spectrum is used to show how strongly or how poorly a compound absorbs the wavelengths of different lights.

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