Introduction to Analytical Chemistry

Lecture Date: January 14, 2013

What is Analytical Chemistry?

Analytical chemistry is the science of obtaining,
processing, and communicating information about
the composition and structure of matter.
In other words, it is the art and science of
determining what matter is and how much of it
exists.

Qualitative: provides information about the identity of

an atomic, molecular or biomolecular species
Quantitative: provides numerical information as to the
relative amounts of species
Definitions from www.acs.org

The Role of Analytical Chemistry

They adapt proven methodologies to new materials/systems or to answer
new questions about their composition.
Analytical chemists work to improve existing techniques to meet the
demands of for faster, cheaper, more sensitive chemical measurements
Analytical chemists research to completely new types of measurements
and are at the forefront of the utilization of major discoveries in fields as
diverse as photonics and implantable microchip sensors.
Analytical chemistry is applied to many branches of science
Medicine
Industry
Environmental
Food and Agriculture
Forensics
Archaeology
Space science

History of Analytical Methods

Classical methods:
Separation via precipitation, extraction or distillation
Qualitative: recognized by color, boiling point, solubility,
taste
Quantitative: gravimetric or titrimetric measurements
Instrumental Methods:
Separation via chromatography and electrophoresis
Qualitative and Quantitative: recognized by their
interaction with radiation (spectroscopy), their mass (mass
spectrometry), their electrical properties, or their
interaction with environment (temperature, humidity)

Modern Instrumental Techniques

Separation Techniques
Gas chromatography
High performance liquid chromatography
Ion chromatography
Super critical fluid chromatography
Capillary electrophoresis
Planar chromatography
Mass Spectrometry
Electron ionization MS
Chemical ionization MS
High resolution MS
Gas chromatography MS
Fast atom bombardment MS
Liquid chromatography MS
Laser MS
Ambient ionization MS

Modern Instrumental Techniques

Spectroscopic techniques
Infrared spectrometry
Raman spectrometry
Nuclear magnetic resonance (NMR)
X-ray spectrometry
Atomic absorption spectrometry
Inductively coupled plasma atomic emission spectrometry
Inductively coupled plasma MS
Atomic fluorescence spectrometry
Ultraviolet/visible spectrometry (CD)
Molecular fluorescence spectrometry
Chemiluminescence spectrometry
X-Ray Fluorescence spectrometry
Electrochemical techniques
Amperometry
Voltammetry
Potentiometry
Conductiometry

Microscopic and surface techniques

Atomic force microscopy
Scanning tunneling microscopy
Auger electron spectrometry
X-ray photon electron spectrometry

Major Steps in Solving an Analytical Problem

1. Understanding and defining the problem, by looking at
the history of the material to be analyzed and background
of the problem
2. Choosing your analytical technique(s) and running the
experiments (or developing a new analytical technique)
3. Data analysis and interpretation, validation of results (if
needed), and reporting of results

1. Understanding and Defining

the Problem

What is it that you want to know?

What accuracy is required?
Is there a time (or money) limit?
How much sample is available?
What is the concentration range of the analyte?
What components of the sample may cause an
interference?
What are the physical and chemical properties
of the sample matrix?
How many samples are to be analyzed?

History of sample and background

of the problem
Background information can originate from many sources
The client and competitors products
Literature searches on related systems
Sample history:
How was the sample collected, transported, and stored?
How was it sampled?
If synthesized, by what synthetic route?
What was the source of the raw materials used to make
the sample?
What analysis has already been performed?

2. Choosing the Analytical Technique

Consider the sample characteristics
Choose an instrument (and ultimately a method) that can
obtain the desired information
Evaluate the performance characteristics of that instrument
and method
Does an entirely new technique need to be developed?

Technique Selection
Analysis type
Quantitative, Qualitative
Location of sample
bulk or surface
Physical state of sample
gas, liquid, solid, dissolved solid, dissolved gas
Amount of Sample
macro, micro, nano,
Fate of sample
destructive, non destructive
Estimated purity of sample
pure, simple mixture, complex mixture
Analyte concentration
major or minor component, trace or ultra trace
Elemental information
total analysis, speciation, isotopic and mass analysis
Qualitative Molecular information
compounds present, polyatomic ionic species, functional
group, structure, molecular weight, physical property

Comparing Two Analytical Techniques:

High pressure Liquid Chromatography (HPLC)
vs. Nuclear Magnetic Resonance (NMR)
HPLC

NMR

L,Ds

L,S,Ds

macro, micro

Ma, Mi

Ma, Mi

pure, simple mixture, complex mixture

Sm,M

P,Sm

Location of sample
bulk or surface
Physical state of sample
gas, liquid, solid, dissolved solid, dissolved gas
Amount of Sample
Estimated purity of sample
Fate of sample
destructive, non destructive
Elemental information

N,D

T,S (ion)

limited

Cp,Io,St

Cp,Fn,St

Ql,Qt

Ql,Qt

total analysis, speciation, isotopic and mass analysis

Molecular information
Compounds present, Polyatomic ionic species,
Functional group, Structural, MW, Physical prop
Analysis type
Quantitative, Qualitative

Components of an Analytical Method

Obtain and store sample
Extract data
from sample

Pretreat and prepare sample

Perform measurement(s)/
experiment and process raw
data (if needed)

Compare results
with standards
Covert data
into information

Apply
Statistics (Quantitative)
Interpret Data (Qualitative)

Transform
information into
knowledge

Present/Report information
in a understandable form

After reviewing results

might be necessary
to modify and repeat
procedure

3. Analyzing Data and Reporting Results

Analytical data analysis takes many forms: statistics,
chemometrics, simulations, empirical interpretation, etc
Analytical results can be reported in
Peer-reviewed papers
Technical reports
Laboratory notebook records
Analytical results can be subject to extreme scrutiny and
can be challenged by other experts

Basis Quantitative Analysis

Precision refers to the reproducibility of analytical results.

When a result is precise, numerical results agree closely.

Precision can be estimated by repeating the measurement
n times (when possible).

Accuracy describes the correctness of a result by its

closeness to an accepted or true value.
Precise, not accurate

Accurate, not precise

Accurate and precise

See pg. 967 of Skoog et al., Principles of Instrumental Analysis, Thomson Brooks/Cole, New York, 2007.

Basis Quantitative Analysis

Selectivity: the extent to which a technique or method can

determine particular analytes under given conditions in
mixtures or matrices, simple or complex, without
interferences from other components.

Also referred to as specificity

Sensitivity: the ability of a technique or method

discriminate between small differences in level of an
analyte

Basis Quantitative Analysis

Limit of detection (LOD): the lowest
amount of an analyte that can be
detected at a known confidence level

Signal-to-noise: ratio of the average

signal to the average level of noise.

Limit of quantitation (LOQ): the range

over which quantitative measurements
can be made (usually the linear range),
often defined by detector dynamic
range

Detector response

Limit of linearity
Slope relates to
sensitivity

LOQ
LOD
Dynamic range

Concentration

Linearity: the degree to which a response of an analytical detector to

analyte concentration/mass approximates a linear function

Dynamic range: range between the LOQ and limit of linearity

Significant Figures

All nonzero digits are significant:

1.234 g has 4 significant figures

Zeroes between nonzero digits are significant:

1002 kg has 4 significant figures

Leading zeros to the left of the first nonzero digits are not significant; such
zeroes merely indicate the position of the decimal point:
0.001 oC has only 1 significant figure

Trailing zeroes that are also to the right of a decimal point in a number are
significant:
0.0230 mL has 3 significant figures

When a number ends in zeroes that are not to the right of a decimal point, the
zeroes are not necessarily significant:
190 miles may be 2 or 3 significant figures

Significant Figures

Addition and subtraction, the result is rounded off so that it has the
same number of digits as the measurement having the fewest decimal
places
100 (no decimal places) + 23.643 (3 places) = 123.643, which
should be rounded to 124 (no places).

In multiplication and division, the result should be rounded off so as to

have the same number of significant figures as in the input value with
the least number of significant figures
3.0 (2 significant figures ) 12.60 (4 significant figures) = 37.8000
which should be rounded to 38 (2 significant figures).

Scientific Notation and Prefixes

0.00000356 M
3.56 x 106 M
3.56 M
or . ppm
Prefixes for SI Units
gigaG
megaM
kilok
decid
centic
millim
micro
nanon
picop
femtof
attoa

109
106
103
10-1
10-2
10-3
10-6
10-9
10-12
10-15
10-18

Working with Numbers: Analytical

Concentrations
Molarity
Moles of solute / L

Parts per Million (ppm)

cppm = mass of solute X 106 ppm
mass of solution
For dilute aqueous solutions whose densities are approximately 1.00
g/mL
1 ppm = 1 mg/L =1 g/mL

Parts per Billon (ppb)

cppb = mass of solute X 109 ppb
mass of solution

or

1 g/L

Basic Statistics

Mean (average) of a population:

Mean (average) of a sample:

The Standard Deviation

The standard deviation indicates the spread of data

The sample standard deviation (for a data set of limited
size) is given by s:

Relative standard deviation (RSD) (%)

See pg. 971-972 of Skoog et al., Principles of Instrumental Analysis, Thomson Brooks/Cole, New York, 2007.

The Gaussian Probability Distribution

If you take a large number

The likelihood of a result

will become lower the

farther away the result is
from the mean

600

Response

of measurements, the
values with be distributed
around the expected value,
or mean

500
400
300
200
100

Measurement (e.g. spectral frequency)

Many physical phenomena studied in analytical chemistry

result in measurements that can be modeled as Gaussian

distributions
See pg. 971-972 of Skoog et al., Principles of Instrumental Analysis, Thomson Brooks/Cole, New York, 2007.

Probability Distributions and Measurement

Confidence Intervals
Consider the following eight results :
2.1

2.3

2.6

2.1

1.9

2.2

1.8

3.8

mean = 2.35 and std deviation = 0.635

The question is, what is the chance that the large value of 3.8
occurred by random chance assuming a Gaussian
distribution?

Confidence Intervals An example

N = 8, mean = 2.35, and s = 0.635

For this example, choose a 95% confidence level.
Use Skoog et al. table A1-5 to obtain t (95% CI, dof
=7)

2.35 (2.36*0.635)/Sqrt[8]
2.35 0.53

Result:

3.8 is outside the range of 2.35 0.53. We

can be 95% confident that the value of 3.8 is from a
different system, etc

Calibration Curves

Measure signal response vs. known

analyte concentration

The data is plotted and fit to a function to

Typically the best fit is linear
y = mx+b

Response

obtain the equation of the best fit and the

uncertainty in the fit.

response = slope [c] + intercept

m is related to the method sensitivity

Measure the sample response to

determine the concentration

Matrix effects must be minimal

Concentration

Linear Least Squares or Linear Regression

Method to minimize the residual of the

experimental values and fitted line

Appendix 1 shows the how to do linear regression

by hand

However, typically this is done with software

Correlation Coefficient (R2 )

A fraction between 0.0 and 1.0
Dimensionless it has no units

Sum of
residuals
associated
with a linear
relationship
SSreg

Sum, of
residuals
associated
with the null
hypothesis
average of the
y values
SStot

If R2 is near 1.0, the regression model

fits the data much better than the null
hypothesis
If the regression model were not much
better than the null hypothesis, R2
would be near zero

Using a Calibration Curve

What is the mole % of
isooctane in the sample
with a peak area of 2.65?
What is the standard
deviation?

sr
stdev
m

1 1 yc y

M N
m 2 S xx

y b y 0.2567
x

m
2.0925
2.65 0.2567

1.14 mol%
2.0925

0.1442 1 1 2.65 12.51 / 5

2.0925 1 5 2.09252 1.145
0.076 mol%

sr= stdev of regression line

M= number of sample measurements
N= number of samples for calibration
y= average peak area of the calibration
Sxx = sum of squares of the deviation for x

The Standard Addition Method

Known quantities of analyte are added to a sample of

unknown concentration

Good for systems with significant matrix effects

(interferences), where selectivity or specificity is lacking

[X] = concentration of analyte

[S] = concentration of standard
I = signal
i = initial, f = final

[x]f

The Standard Addition Method: An Example

Example: atomic absorbance
measurements of a Zn spectral
line for determination of Zn2+
Three standard additions are made
to the actual sample, which is also
analyzed (four samples)

Data analyzed by linear regression

0.6
0.5

(A = absorbance, [c] = concentration)

0.4

Solve for [X]f , where A = 0

Zn2+ in sample =1.11 ppm

Absorbance

A = 0.179[c] + 0.199

0.3
0.2
0.1
0
0.0

0.5

1.0
1.5
2.0
Zinc Concentration (ppm )

Example from H. A. Strobel and W. R. Heineman, Chemical Instrumentation: A Systematic Approach,

Wiley: New York, 1989, p. 393.

2.5

Non-Linear Fitting
Function

Form

Example

Logarithmic

S=a+blnC

Nernst Equation

Exponential

S=aebC

Healy's model for

immunoassay

Power

S=a+bCn

Kohlrauschs Law

Polynomial

S=a+bC2+ cC3...

Immunometric
assays

The Nernst equation is an equation that can be used (in conjunction with other
information) to determine the equilibrium reduction potential of a half-cell in an
electrochemical cell.
Kohlrauschs Law states that the conductivity of a dilute solution is the sum of
independent values: the molar conductivity of the cations and the molar
conductivity of the anion. The law is based on the independent migration of ions.

Simple Chemical Tests

While most of this class is focused on instrumental

methods, a very large number of simple chemical tests
have been developed over the past ~300 years

Examples:
Barium: solutions of barium salts yield a white precipitate with 2
N sulfuric acid. This precipitate is insoluble in hydrochloric acid
and in nitric acid. Barium salts impart a yellowish-green color to
a non-luminous flame that appears blue when viewed through
green glass.
Phosphate: With silver nitrate TS, neutral solutions of
orthophosphates yield a yellow precipitate that is soluble in 2 N
nitric acid and in 6 N ammonium hydroxide. With ammonium
molybdate TS, acidified solutions of orthophosphates yield a
yellow precipitate that is soluble in 6 N ammonium hydroxide.
Examples are from US Pharmacopeia and National Formulary USP/NF

A Colormetric Test for Mercury

A modern example of a
spot test: a test for Hg2+
developed using DNA and
relying on the formation of
a thymidine-Hg2+thymidine complex

LOD = 100 nM (20 ppb) in

aqueous solution

Linearity from the high

nanomolar to low micromolar
range

Selective for Hg2+ and

insensitive to Mg2+, Pb2+, Cd2+,
Co2+, Zn2+, Ni2+, and other
metal ions
Angew. Chem. Int. Ed., DOI: 10.1002/anie.200700269
http://pubs.acs.org/cen/news/85/i19/8519news6.html

Further Reading

Optional Reading:
Skoog et al. Chapter 1 and Appendix 1
Skoog et al. Chapters 6 and 7 (Spectroscopy)