READING SUPPLEMENT

INSTRUMENTAL METHODS OF ANALYSIS: AN INTRODUCTION

Analytical methods are usually classified as being either classical or instrumental. In classical methods, also
known as wet-chemical methods, the measurement depends on the chemical properties of the sample. A reagent is
made to react completely with the analyte and the relationship between the measured signal and the analyte
concentration is determined by chemical stoichiometry. Separation of components of interest in a sample is by
precipitation, extraction or distillation. Qualitative analyses of the separated components are performed based on
colors, boiling or melting points, solubility in different solvents, odors, optical activities or refractive indices of products
formed upon treatment with reagents. Quantitative analyses is done using gravimetric or volumetric measurements.

Instrumental methods of analysis are based on the measurement of physical or chemical properties of the
analyte. Separation of components of complex mixtures are performed using chromatography and electrophoretic
techniques. Quantitative analyses is based on measurements of physical properties like conductivity, electrode
potential, light absorption or emission, mass-to-charge ratio and fluorescence.

Classical Instrumental
More suitable for analysis of major constituents Ability to perform trace analysis
Used to certify analytical standards Most are multi-channel techniques
Generally cheaper Shorter analysis time
More accurate and precise Amenable to automation
More robust and less susceptible to environmental Large numbers of samples may be analyzed quickly
fluctuations Less skill and training required

Table 1. Types of Instrumental Methods of Analysis

Classification Characteristic Properties Instrumental Methods
A. Spectrometric Methods Emission of radiation Emission spectroscopy (X-ray, UV, visible, electron,
Auger), fluorescence, phosphorescence and
luminescence (X-ray, UV and visible)
Absorption of radiation Spectrophotometry and photometry (X-ray, UV, IR);
photoacoustic spectroscopy; nuclear magnetic
resonance and electron spin resonance
spectroscopy
Scattering of radiation Turbidimetry; nephelometry; Raman spectroscopy
Refraction of radiation Refractometry; interferometry
Diffraction of radiation X-ray and electron diffraction methods
Rotation of radiation Polarimetry; optical rotary dispersion; circular
dichroism
B. Chromatographic Methods High Performance Liquid Chromatography
Gas Chromatography
C. Electrochemical Methods Electrical potential Potentiometry
Electrical charge Coulometry
Electrical current Amperometry; polarography
Electrical resistance Conductometry
D. Miscellaneous Methods Mass-to-charge ratio Mass spectrometry
Thermal characteristics Thermal gravimetry and titrimetry; differential
scanning calorimetry; differential thermal analyses;
thermal conductometric methods
Radioactivity Activation and isotope dilution methods
ust-gccastro-072014 Page | 1
 READING SUPPLEMENT

Analytical Instruments
An instrument for chemical analysis converts information about the physical or chemical characteristics of the
analyte to information that can be manipulated and interpreted by a human. Thus, an analytical instrument can be
viewed as a communication device between the system under study and the investigator. To retrieve the desired
information from the analyte, it is necessary to provide a stimulus, which is usually in the form of electromagnetic,
electrical, mechanical, or nuclear energy. The stimulus elicits a response from the system under study whose nature
and magnitude are governed by the fundamental laws of chemistry and physics. The resulting information is contained
in the phenomena that result from the interaction of the stimulus with the analyte. (Figure 1.1)

TRANSDUCER
SIGNAL READ-OUT
Stimulus OR
PROCESSOR DEVICE
DETECTOR
COMPONEN
T
Energy source Chemical
system Analytical signal

Electrical signal
SIGNAL GENERATOR
Figure 1. Diagram showing the general components of an instrumental measurement

The signal generator consists of the chemical system interacting with the stimulus from the energy source
which results in the production of an analytical signal reflecting the presence and usually the concentration of the
analyte. The transducer or detector transforms the analytical signal produced by the signal generator into an electrical
signal. The signal processor modifies and “cleans up” the electrical signal to make it more convenient to interpret.
Finally, the read-out device converts the electrical signal to a form usable to the analyst.

Table 2. Examples of Instrument Components

Instrument Energy Source Analytical Information Input Data Signal
(stimulus) Information Sorter Transducer Domain of Processor /
Transduced Readout
Information
Photometer Tungsten Attenuated Filter Photodiode Electrical Amplifier/
lamp light beam current LED display
Atomic Inductively UV or visible Monochromator Photomultiplier Electrical Amplifier/
emission coupled radiation tube current Digital
spectrometer plasma display
Coulometer Direct-current Charge Cell potential Electrodes Time Amplifier/
source required to Digital
reduce or timer
oxidize
analyte
pH meter Sample/glass Hydrogen ion Glass electrode Glass-calomel Electrical Amplifier/
electrode activity electrodes voltage Digital
display
Mass Ion source Mass-to- Mass analyzer Electron Electrical Amplifier/
spectrometer charge ratio multiplier current Computer
system
ust-gccastro-072014 Page | 2
 READING SUPPLEMENT

Calibration of Instrumental Methods

Calibration determines the relationship between the analytical response and the analyte concentration.
Usually this is determined by the use of chemical standards. There are several types of calibration procedures:
(1) Comparison with standards, (2) External standard calibration, (3) Standard addition method, and (4) internal
standard method.
Comparison with standards may be done by direct comparison or titrations. In direct comparison, the property
of the analyte is compared with standards such that the property being tested matches the standard. For example, in
early colorimeters, the color produced as the result of a chemical reaction of the analyte was compared with the color
produced by reaction of standards. If the concentration of the standard was varied by dilution, for example, it was
possible to obtain a fairly exact color match. The concentration of the analyte was then equal to the concentration of
the standard after dilution. Titrations are among the most accurate of all analytical procedures. In a titration, the
analyte reacts with a standardized reagent (titrant) in a reaction of known stoichiometry. Usually the amount of titrant
is varied until chemical equivalence is reached, as indicated by the color change of a chemical indicator or by the
change in an instrument response. The amount of the standardized reagent needed to achieve chemical equivalence
can then be related to the amount of analyte present.
An external standard is prepared separately from the sample. External standards are used to calibrate
instruments and procedures when there are no interference effects from matrix components in the analyte solution. A
series of such external standards containing the analyte in known concentrations is prepared. Ideally, three or more
such solutions are used in the calibration process. Calibration is accomplished by obtained the response signal
(absorbance, peak height, peak area) as a function of known analyte concentration. A calibration curve is prepared by
plotting the data or by fitting them to a suitable mathematical equation, such as the slope-intercept form used in the
method of linear least squares. The response signal is then obtained for the sample and used to predict the unknown
analyte concentration from the calibration curve or best-fit equation.
Standard-addition methods are particularly useful for analyzing complex samples in which the likelihood of
matrix effects is substantial. One of the most common form of standard-addition involves adding one or more
increments of a standard solution to sample aliquots containing identical volumes. This process is often called “spiking”
the sample. Each solution is then diluted to a fixed volume before measurement.
An internal standard is a substance that is added in a constant amount to all samples, blanks, and calibration
standards in an analysis. Calibration involves plotting the ratio of the analyte signal to the internal-standard signal as a
function of the analyte concentration of the standards. This ratio for the sample is then used to obtain their analyte
concentration from a calibration curve.

Signals and Noise

Every analytical measurement is made up of two components. One component, the signal, carries information
about the analyte that is of interest to the scientist. The second, called noise, is made up of extraneous information
that is unwanted because it degrades the accuracy and precision of an analysis and also places a lower limit on the
amount of analyte that can be detected.
Chemical analyses are affected by two types of noise: chemical noise and instrumental noise. Chemical noise
arise from a host of uncontrollable variables that affect the chemistry of the system being analyzed. Examples include
undetected variations in temperature or pressure that affect the position of chemical equilibria, fluctuations in relative
humidity that cause changes in the moisture content of samples, vibrations that lead to stratification of powdered
solids, changes in light intensity that affect photosensitive materials, and laboratory fumes that interact with samples
or reagents. Instrumental noise is associated with each component of an instrument, i.e. the source, input transducer,
signal-processing elements, and the output transducer. Noise that is observed is usually a complex composite of
several sources of noise that cannot be fully characterized. An example is environmental noise which is a composite of
different forms of noise that arise from surroundings.

ust-gccastro-072014 Page | 3
 READING SUPPLEMENT

Selecting an Analytical Method

There is an enormous array of tools for carrying out chemical analyses. Selection of a suitable method may be
guided by answering the following questions:
(1) What accuracy is required?
(2) How much sample is available?
(3) What is the concentration of the analyte?
(4) What components of the sample might cause interference?
(5) What are the physical and chemical properties of the sample matrix?
(6) How many samples are to be analyzed?
Other considerations in method choice are speed, ease and convenience, skill required of operator, cost and
availability of equipment and per-sample cost.

References:
Harris, DC (2005). Exploring Chemical Analysis 3rd Ed. W.H. Freeman and Company: New York
Skoog, DA, Holler, FJ and Crouch, SR (2007). Principles of Instrumental Analysis 6th Ed. Thomson Brooks/Cole: USA.
pp.1-24
Tissue, BM (2013). Basics of Analytical Chemistry and Chemical Equilibria. John Wiley & Sons, Inc.: New Jersey.
https://facultystaff.richmond.edu/~rdominey/301/local/Intro_Instrum_Analysis.pdf (accessed July 01, 2014)

ust-gccastro-072014 Page | 4