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Lab Report

This lab report describes how students determined the concentration of blue dye in four unknown solutions by generating a standard curve using a spectrophotometer. Students created solutions of known blue dye concentrations and measured their absorbance. They plotted the data in Excel to generate a standard curve with an equation. The students then measured the absorbance of the four unknown solutions and plugged those values into the standard curve equation to calculate the blue dye concentration in each unknown solution. The concentrations determined were 9.19, 3.695, 0.551, and 5.07 μg/ml for Jagged Ice, Cool Blue, Mixed Berry, and Mountain Blast, respectively.

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Clayton VerBerkmös
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433 views3 pages

Lab Report

This lab report describes how students determined the concentration of blue dye in four unknown solutions by generating a standard curve using a spectrophotometer. Students created solutions of known blue dye concentrations and measured their absorbance. They plotted the data in Excel to generate a standard curve with an equation. The students then measured the absorbance of the four unknown solutions and plugged those values into the standard curve equation to calculate the blue dye concentration in each unknown solution. The concentrations determined were 9.19, 3.695, 0.551, and 5.07 μg/ml for Jagged Ice, Cool Blue, Mixed Berry, and Mountain Blast, respectively.

Uploaded by

Clayton VerBerkmös
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Biology L113 09/08/08 Determining Concentration by Using a Standard Curve Methods In the Standard Curve for F D and C Blue

#1 laboratory our group learned a number of important processes and utilized different methods in order to determine the unknown concentrations of several liquids. As described in L113 Laboratory Manual (Bonner et al., 2008), we used dilution, spectrophotometry, and MS excel graphing to make this determination. More specifically, we began the experiment by creating seven test tube samples of known concentrations of blue dye by dilution using micropipettes. We labeled each test tube, with the exception of the test tube labeled 0 in which there was no dye (the blank), then started with a set volume of concentrated dye (20, 60, 150, 300, 600, and 900 l, respectively) in which we added water (2.98, 2.94, 2.85, 2.7, 2.4, and 2.1 ml, respectively) in order to have a final volume of 3.0 ml. From this information, we calculated the final concentration and dilution factor for each test tube as instructed on page B-3 of the Laboratory Manual (Bonner et al., 2008) and recorded them on page B-6. Once we prepared these solutions, we set the spectrophotometer to 650 nm and recorded the absorbency of each sample. We did an addition trial for each concentration to ensure that the data collected was as accurate as possible. Had Trial 2 data deviated 5% or more from Trial 1 data, we would have run the experiment a third or even forth time. We then averaged the absorbencies of the two trials and plotted this average in Microsoft Excel. Once we plotted the points we applied a trend-line (the standard curve) to the graph along with the equation of the line and R2-value. A good indicator of the accuracy of data is if the R2value is close to 1.

The next part of the lab required us to analyze four different solutions: Gatorade Cool Blue, Gatorade Blue Mountain, Powerade Jagged Ice, and Powerade Zero Mixed Berry. Each of these solutions contained an unknown amount of blue dye that had to be determined. We analyzed each solution in the spectrophotometer in the same fashion as the analysis for the standard curve. We then recorded the resulting absorbencies. Because each of the recorded

absorbencies fell within the range of the standard curve there was no need to dilute any of the unknown solutions. Finally we substituted the absorbency reading for the four unknown samples for the y value in the equation of the line from the standard curve, which then allowed us to solve for x (which represents the concentration of dye in g/ml). Summary The purpose of this lab was to determine the concentration of four unknown solutions. Generally speaking, the most efficient way to find unknown concentrations is to generate a standard curve using a spectrophotometer, analyze the absorbencies of unknown solutions, and then substitute the measured absorbencies for the y-value in the equation-of-a-line from the standard curve. A spectrophotometer works by passing light through a liquid sample and determining how much of that light was absorbed by the solution. The absorbencies determined in this way for several stock solutions were plotted on a scatter graph in MS Excel in order to create a standard curve. We then used this standard curve to compare to the absorbencies of the unknown solutions by plugging in the absorbency for the y-value and solving for x. The concentrations for Jagged Ice, Cool Blue, Mixed Berry, and Mountain Blast were 9.19, 3.695, 0.551, and 5.07 g/ml, respectively. The graph of the standard curve is attached. This shows that the lighter in color the solution was that was being tested, the lower the concentration of blue dye.

Literature Cited Bonner, Jose, James Hengeveld, Richard Holdeman, William Ruf, and Evelyn Rynkiewicz. 2008. Biology L113 Laboratory Manual. Department of Biology, Indiana University, Bloomington, IN.

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