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Eleanor Calibration Curves_emw

# Eleanor Calibration Curves_emw - Calibra'onCurves...

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Calibra’on Curves Eleanor Waxman 28 and 30 September 2011 [email protected]

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Plan 1. Regular Calibra’on Curves (Wed) 2. Standard Addi’on (Wed/Fri) 3. If ’me, in class analysis example (Fri) Please bring your computer. To prep for this, please download “Data for Friday, 30 September.xlsx” or “Data for Friday, 30 September_xlscompa’ble.xls” and have it loaded.
What is a calibra’on curve? A plot of instrument response versus analyte concentra’on. y = 0.9948x ‐ 0.0214 R² = 0.9981 ‐0.02 0.08 0.18 0.28 0.38 0.48 0.58 0 0.2 0.4 0.6 0.8 Absorbance(arbitraryunits) Concentra2on (M) Sample Calibra2on Curve

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How do you make one? For this example, let’s add food coloring to 50 mL of water. Standard: Solu’on with known amount of analyte
Blanks Blank: Solu’on with all reagents and solvents, but no analyte. In the previous example, it would be the same type of water used to make the standards. Blank Standards

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Discussion Ques’on 1 What would be an appropriate blank for studying proteins in a carbonate buffer? Next semester, there’s a lab looking at tartrazine (yellow dye) in Mountain Dew. What would be an appropriate blank?
How do you make one? Drops Added Absorbance (532 nm) 0 0.01 10 0.18 20 0.41 30 0.60 40 0.79 50 1 0 0.2 0.4 0.6 0.8 1 0 10 20 30 40 50 Absorbance Drops of Food Coloring Added Absorbance versus Drops Added

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How do you make one? Drops Added Absorbance (532 nm) 0 0.01 1 0.02 2 ‐0.01 3 0.01 5 0.11 10 0.18 15 0.29 20 0.41 25 0.52 30 0.60 35 0.71 40 0.79 45 0.92 50 1 55 0.98 60 0.99
This is a scafer plot of the data. ‐0.2 0 0.2 0.4 0.6 0.8 1 1.2 0 10 20 30 40 50 60 70 Absorp2on(unitless) Drops Added Absorbance of Green Food Coloring at 532 nm

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Clicker Ques’on 1 ‐0.2 0 0.2 0.4 0.6 0.8 1 1.2 0 10 20 30 40 50 60 70 Absorp2on(unitless) Drops Added Absorbance of Green Food Coloring at 532 nm A B C Which region(s) would you use to calculate your calibra’on curve? A: A B: B C: C D: A and B E: A, B, and C
Clicker Ques’on 1 Analysis ‐0.2 0 0.2 0.4 0.6 0.8 1 1.2 0 10 20 30 40 50 60 70 Absorp2on(unitless) Drops Added Absorbance of Green Food Coloring at 532 nm A B C Which region(s) would you use to calculate your calibra’on curve? B is the only linear region. This is the only part we can easily fit a line to.

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Clicker Ques’on 2 ‐0.2 0 0.2 0.4 0.6 0.8 1 1.2 0 10 20 30 40 50 60 70 Absorp2on(unitless) Drops Added Absorbance of Green Food Coloring at 532 nm A B C What is going on in Region A?
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Eleanor Calibration Curves_emw - Calibra'onCurves...

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