ChE 408 – Engineering Experimental Design

In-Class Project

Calibration Curve for Spectrophotometric Determination of Crystal Violet Dye Concentration

 

Here is a page from a laboratory notebook.  Measurements were made to develop a calibration curve so that the concentration (in moles per liter) of an aqueous dye solution may be determined by measuring the UV absorbance of the solution.  Using the data provided, develop an equation that will allow someone to calculate the concentration of a CVD solution given its absorbance.  Evaluate the validity of your equation.  Then determine the concentrations of solutions whose absorbances are 0.5, 1.0, and 1.5.  (For now, use the confidence limits on the adjustable parameters to estimate the uncertainty in the concentration.  Later in the course, we will learn a more correct way to estimate the uncertainty on values predicted using a regression model.)

 

 


Laboratory Notebook

 

Calibration data for aqueous crystal violet dye (CVD), UV absorbance at 492 nm. 

Absorbance = -log10(light transmitted / incident light); 100 % light transmitted gives an absorbance of zero.

 

CVD stock solution

0.3479 ± 0.0010 g CVD (formula weight 408 g/mole) placed in 1 L volumetric flask; diluted with water to 1000 ± 1 ml.

 

Determination of UV absorbance of standard solutions

Diluted measured volume of CVD stock solution with water in volumetric flask.  Placed resulting dilute solution in UV cuvette and measured absorbance at 492 nm. 

 

Run #

CVD stock solution added

ml

Total solution volume

ml

Absorbance

1

0

 

0.00021

2

8.5 ± 0.1

1000 ± 1

0.65731

3

Repeat of run #2

 

0.6635

4

8.5 ± 0.1

1000 ± 1

0.62227

5

10.0 ± 0.1

1000 ± 1

0.7552

6

25.0 ± 0.1

1000 ± 1

1.84568

7

10.0 ± 0.1

500 ± 1

1.43606

8

Repeat of run #5

 

0.74222

9

Repeat of run #2

 

0.62371

10

13.5 ± 0.5

500 ± 1

1.93544

11

15.0 ± 0.5

1000 ± 1

1.07576

12

5.9 ± 0.2

1000 ± 1

.41681