bicol university - weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · a number of...

16
BICOL UNIVERSITY College of Science Department of Chemistry CHEM 22 INSTRUMENTAL METHODS OF ANALYSIS LAB EXERCISE 2 Ver. 1.0 α 20151124

Upload: others

Post on 18-Aug-2020

3 views

Category:

Documents


0 download

TRANSCRIPT

Page 1: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

BICOL UNIVERSITY College of Science

Department of Chemistry

CHEM 22 INSTRUMENTAL METHODS OF ANALYSIS

LAB EXERCISE 2 Ver. 1.0 α 20151124

Page 2: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

2 http://buchem.weebly.com michaelvmontealegre

Page 3: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

Exercise 1: Using Excel Functions:

We have already seen how Excel can be used to perform simple calculations by typing equations into cells within the spreadsheet. Obviously, we would want to perform calculations using basic mathematical functions and constants, such as sin x, log y, or π. In this section, we will use Excel's built-in functions to perform elementary calculations. Functions are entered in cells as part of an equation; remember that equations in cells always begin with an “=” sign. We will also use the techniques learned in the preceding exercises to create a series of values, fill cells in the same column with the same formula, and perform calculations using both absolute and relative referencing.

The LOG and LN functions Provide the plots using log and ln, by following the instructions below.

A number of analytical techniques require measurement data to be transformed in some manner before a calibration graph can be constructed. One common example is potentiometry, in which the measured response (electrode potential, E Volts) is related to the logarithm of the corresponding ion activity (a) or concentration (C) via the Nernst equation. For a simple electrochemical cell involving a single metal species being reduced at the indicating (measured) electrode, the equation for the reaction for Mn+ + ne– can be written as:

1. Open a new Excel™ worksheet and enter the text C(mol/L) in cell A1, then create a series of concentration values from 0.00 to 1.00 mol/L in increments of 0.05 mol/L starting in cell A2.

2. Enter the text ln C in cell B1, then enter the equation =LN(A2) in cell B2; use Edit→Fill→Series... or click-and-drag the small square on the bottom-right corner of the selected cell to fill cells B2 to B20 with the values of ln(C) corresponding to the values of C in column A.

Before calculating values of the electrode potential, we will first make life a little easier by setting up a table of constants, and pre-calculating the value of the slope term, RT/nF. In order to avoid problems, we will use absolute referencing when using these values.

spreadsheet 3

Page 4: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

1. Create a table of the following constants at a convenient place within the worksheet, such as cells E1 to F6: E°(Cu2+) = 0.337 V,R = 8.31451 J mol-1 K-1, T = 298.15 K, n = 2, F=96485 C mol-1.

2. Create an entry for the term RT/nF, and enter the appropriate equation in the adjacent cell. Remember to use brackets to keep the terms in the denominator together!

3. Format the cells to show the correct number of significant figures and/or decimal places.

4. Enter the text E (V) in cell C1, create an equation for E in cell C2, and fill the cells down to complete the table of calculated electrode potentials.

You can now plot a graph of the electrode potential for a Cu|Cu2+ electrode as a function of (i) concentration and (ii) ln C. Don‘t forget to format the graph properly. You can use the options in the Scale tab of the Format Axis dialog to position the y-axis on the left-hand side of the graph.

2nd Plot: Change the formula to the more common "log" format by calculating 2.3026RT/nF, and changing column B to log C by using Excel's LOG() function.

4 http://buchem.weebly.com michaelvmontealegre

Page 5: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

spreadsheet 5

Page 6: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

Exercise 2: Calibration and Trendlines:

Most analytical chemistry measurements involve calibration functions that can be described using a straight line. Excel has a feature that allows you to easily display a linear trendline on your graphs, which is the best-fit straight line through your data. This feature also allows you to view the equation of the best-fit line, as well as the correlation coefficient; both of these are determined using the mathematical technique of linear regression analysis. The trendline feature provides a quick test of the linearity of your calibration data. A more complete treatment of linear regression will be provided in a later section.

A calibration curve is an equation that permits us to calculate a desired experimental result in terms of another. In the simplest form, this is given as the equation of a straight line where the x-value is the input (usually concentration) and the y-value is the output (usually the measured instrument response). As we saw in the preceding example, the relationship between response and concentration is not always linear, although it is usually possible to linearise the equation – that is, transform the data into a form that is linear, such as E versus log C.

Part 1: Specimen Data

In this section, we will use the following calibration data for a fluorescence spectrophotometry experiment. The data is taken from Miller and Miller. Enter the following data in the second two columns of a new Excel spreadsheet. Column B should contain the concentrations and column C the fluorescence intensities. This makes it easier to get the required plot in Excel. Remember that we are determining the regression of y on x – i.e. the dependence of the measured intensity (dependent variable) on the standard concentration (independent variable).

Concentration / pg mL-1 Intensity 0.00 2.1 2.00 5.0 4.00 9.0 6.00 12.6 8.00 17.3

10.00 21.0 12.00 24.7

6 http://buchem.weebly.com michaelvmontealegre

Page 7: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

Don’t forget to include headings for each column in the spreadsheet, and format the cells to display the correct number of decimal places. Once the data is entered and formatted correctly, select both columns then choose Insert→Chart and insert an XY Scatter Plot without connecting lines or interpolated curves.

Hint: Put the independent variable (x) in the first column, and the dependent variable (y) in the second.

Part 2: Adding a trendline

Complete the fields in the various parts of the Chart Wizard to format and produce your calibration graph, remembering to include a title and labels for both axes.

Click on any point on the graph, then choose Chart→Add Trendline... to bring up the Add Trendline dialog. Alternatively, you can right-click on one of the points to access the same menu function.

spreadsheet 7

Page 8: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

Make sure that a linear trendline is selected (but note the other types available!), then click on the Options tab.

Check the options for Display equation on chart and Display R-squared value on chart, then click on the OK button. This will add both the equation for the best-fit straight line through the data and the squared value of the product-moment correlation coefficient (R2) to the chart.

Once you have the equation and R2 on the chart, you will need to format the numbers in this display in order to obtain sufficient figures for accuracy in any further calculations. To do this, either right-click (Mac: cmd-click) on the equation and choose Format Data Labels... from the contextual menu, or left-click on the equation and choose Format→Selected Data Labels.... The Number tab of the resulting dialog box will allow you to set the number of decimal places used; you will need to display at least 5 decimal points on your chart.

Part 3: Finished Chart & Explanation

When you are done, your calibration graph should look like the following.

8 http://buchem.weebly.com michaelvmontealegre

Page 9: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

The best-fit straight line (the line of regression of y on x) is calculated assuming that all errors are in the measured (y) values, not in the concentration (x) values.)

This regression line minimizes the distance in the y direction between the line and the individual points, and passes through the centroid of the data (mean x & y values)

R is a measure of correlation, not linearity. As a result, it can be misleading. You should always look at the graph with the best-fit straight line rather than relying on R alone when evaluating linearity.

spreadsheet 9

Page 10: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

Exercise 3: Standard Addition and Internal Standards See handout for Standardization for formula/details of calculations. 1. To determine the concentration of analyte in a sample, a standard

additions was performed. A 5.00-mL portion of the sample was analyzed and then successive 0.10-mL spikes of a 600.0ppb standard of the analyte were added, analyzing after each spike. The following results were obtained

Volume of Spike

(mL) Signal

(arbitrary units) 0.00 0.10 0.20 0.30

0.119 0.231 0.339 0.442

a. Construct an appropriate standard additions calibration curve. Add the

trendline on the plot with the y-intercept equation and R2 b. Determine the value of the slope and the y-intercept using the spreadsheet

function. c. Determine the concentration of analyte in the original sample

10 http://buchem.weebly.com michaelvmontealegre

Page 11: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

2. Troost and Olavesen investigated the application of an internal standardization to the quantitative analysis of polynuclear aromatic hydrocarbons. The following results were obtained for the analysis of the analyte phenanthrene using isotopically labeled phenanthrene as an internal standard

a. Determine the standardization relationship by a linear regression. calculate the slope and y-intercept (get the average of SA/SIS for each reading before you calculate).

b. What is the value of K. c. Calculate CA given that CIS is 220ppm and SA/SIS of the sample

is 1.934.

spreadsheet 11

Page 12: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

REFERENCE TABLES These tables are from the old Microsoft Excel (2007?) de Levie, 2004, please check the updated functions from the help menu of the software you are using.

12 http://buchem.weebly.com michaelvmontealegre

Page 13: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

spreadsheet 13

Page 14: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

14 http://buchem.weebly.com michaelvmontealegre

Page 15: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

spreadsheet 15

Page 16: BICOL UNIVERSITY - Weeblybuchem.weebly.com/uploads/1/9/3/5/193546/2015-2016... · A number of analytical techniques require measurement data to be ... De Levie, Robert. How to use

Book Reference: De Levie, Robert. How to use excel® in analytical chemistry and in general

scientific data analysis. Cambridge University Press, UK. 2004 Internet Source

http://www.chem.utoronto.ca/coursenotes/analsci/stats/Functions.html

16 http://buchem.weebly.com michaelvmontealegre