The Vocabulary of Analytical Chemistry (cont.

Developing the procedure

Compensating for interferences
Calibration Sampling Validation

Compensating for interferences

A methods accuracy depends on its selectivity for the analyte. Potential interferents may be present in the sample itself or in the reagents used during the analysis. how to minimize these two sources of interference ??? 1) When the sample is free of interferent

Stotal = SA + Sreag = knA + Sreag Stotal = SA + Sreag = kCA + Sreag

the total signal, Stotal the signal due to the analyte, SA the signal due to interferents in the reagents, Sreag

measure the signal for a method blank

Compensating for interferences

Method blank: A sample that contains all components of the matrix except the analyte. A method blank also is known as a reagent blank. When the sample is a liquid, or is in solution, we use an equivalent volume of an inert solvent as a substitute for the sample. Sreag SA

Compensating for interferences

2) an interferent is a part of the samples matrix If the identity and concentration of the interferent are known OK! add to the reagent blank Not common!!! Common: the identity or concentration of matrix interferents is not known, and their contribution to Stotal is not included in Sreag . Instead, the signal from the interferent is included as an additional term
total

total

find a method for separating the analyte and interferent by removing one from the sample
Stotal,1 = kA,1nA + kI,1nI + Sreag,1 Stotal,2 = kA,2nA + kI,2nI + Sreag,2

CALIBRATION
Calibration ensures that the equipment or instrument used to measure the signal is operating correctly by using a standard known to produce an exact signal.
Standardization is the process of experimentally determining the relationship between the signal and the amount of analyte (the value of k by measuring the signal for one or more standard samples, each containing a known concentration of analyte). Several standards with different concentrations of analyte are used the result is best viewed visually by plotting Stotal versus the concentration of analyte in the standards Calibration curve.

how a methods signal changes with respect to the amount of analyte

Example of a calibration curve. The filled circles () are the individual results for the standard samples and the line is the best fit to the data determined by a linear regression analysis.

Analytical Standards
Primary Standards: a reagent extremely pure, stable, no waters of hydration, and a high molecular weight.
Ex:-sodium carbonate: Na2CO3, mol wt. = 105.99 g/mol

-potassium hydrogen iodate: KH(IO3)2, mol wt. = 389.92 g/mol

-potassium dichromate: K2Cr2O7, mol wt. = 294.19 g/mol

(see Appendix 2)
Secondary Standards: a standard prepared in the laboratory for a specific analysis. It is usually standardized against a primary standard.

Analytical Standards
Other reagents: Preparing a standard often requires additional substances that are not primary or secondary reagents. Preparing a standard solution: require a suitable solvent, and additional reagents to adjust the standards matrix.

reagent grade chemicals conforming to standards set by the American Chemical Society should be used.
The label on the bottle of a reagent grade chemical lists either the limits for specific impurities, or provides an assay for the impurities. (We can improve the quality of a reagent grade chemical by purifying it, or by conducting a more accurate assay).

Examples of typical packaging labels for reagent grade chemicals. Label (a) provides the manufacturers assay for the reagent, NaBr. Note that potassium is fagged with an asterisk (*) because its assay exceeds the limits established by the American Chemical Society (ACS). Label (b) does not provide an assay for impurities, but indicates that the reagent meets ACS specifications. An assay for the reagent, NaHCO3 is provided.

Preparing Standard Solutions

a series of standards, each with a different concentration of analyte If the range of concentrations is limited to one or two orders of magnitude: each solution is best prepared by transferring a known mass or volume of the pure standard to a volumetric flask and diluting to volume.

Preparing Standard Solutions

with larger ranges of concentration: standards are best prepared by a serial dilution from a single stock solution. Example: To carry out a serial "ten-fold" dilution

Add 9 mL of solvent to each test tube. To the first test tube, add 1 mL of the stock solution. Mix or vortex. Add 1 mL of this solution to the second test tube and mix. Repeat until you have reached your desired concentration.

Serial dilutions must be prepared with extra care because an error in preparing one standard is passed on to all succeeding standards !!!

Example: Use an analytical balance to weigh a standard for making a calibration curve. + Take note the exact number appearing on the balance because you will use this value to calculate the series of standards concentration. For example, you have the number of 0.0209 g appearing on the balance just take note as it is (although your intentional number was 200 ppm). It doesnt matter if you take the number of exact 200 ppm or 209 ppm because both of them are in the range of the instruments detection. + Usually use a volumetric flask for stock solution: Dissolve 0.0209 g of the standard in 100 mL volumetric flask by solvent A 0.0209 g/ 100 mL = 20.9 mg/ 100 mL = 20900 g/ 100 mL = 209 g/ mL + 209 ppm 20.9 2.09 From 209 ppm to 104.5 ppm
10 fold dilution: 10 fold dilution: 1mL(209 ppm)+9mL 1mL(20.9 ppm)+9mL solvent A solvent A 10 fold dilution: 10 fold dilution: 1mL(104.5 1mL(10.45 ppm)+9mL solvent A ppm)+9mL solvent A

104.5

10.45

1.045

2 fold dilution: 5mL (209 ppm) + 5mL solvent A From 104.5 ppm to 52.25 ppm 2 fold dilution: 5mL (104.5 ppm) + 5mL solvent A

52.25

5.225

0.5225

6 points for the calibration curve in this case

the most desirable situation since the methods sensitivity remains constant throughout the analytes concentration range.

Find out analytes concentration = x from its signal y and the equation y=1020.38x-3.50

Correlation Coefficient, r

where n is the number of pairs of data

measures the strength and the direction of a linear relationship between two variables

-1 < r < +1
0.90 < r < 0.95 : linearity

0.95 < r < 0.99 : good linearity

0.99 < r : excellent linearity

Correlation Coefficient, r
Positive correlation: If x and y have a strong positive linear correlation, r is close to +1. An r value of exactly +1 indicates a perfect positive fit. Positive values indicate a relationship between x and y variables such that as values for x increase, values for y also increase.

Negative correlation: If x and y have a strong negative linear correlation, r is close to -1. An r value of exactly -1 indicates a perfect negative fit. Negative values indicate a relationship between x and y such that as values for x increase, values for y decrease.

No correlation: If there is no linear correlation or a weak linear correlation, r is close to 0. A value near zero means that there is a random, nonlinear relationship between the two variables. Note that r is a dimensionless quantity; that is, it does not depend on the units employed. A perfect correlation of 1 occurs only when the data points all lie exactly on a straight line. If r = +1, the slope of this line is positive. If r = -1, the slope of this line is negative.

Coefficient of Determination, r 2 or R2
The coefficient of determination (R2) is a measure of the fraction of the total variation in y that can be explained by the linear relationship between x and y. The closer R2 is to unity, the better the linear model explains the y variations.

if r = 0.922, then r

= 0.850, which means that

85% of the total variation in y can be explained by the linear relationship between x and y (as described by the regression equation). The other 15% of the total variation in y remains

unexplained.