Instruments for Analysis

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, as illustrated in the figure below. 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.

Generally, instruments for chemical analysis comprise just a few basic components, some of
which are listed in Table 1-2. To understand the relationships among these instrument
components and the flow of information from the characteristics of the analyte through the
components to the numerical or graphical output produced by the instrument, it is instructive
to explore how the information of interest can be represented and transformed.
 Any measurement process can be represented as a series of interdomain
conversions. For example, Figure 1-3 illustrates the measurement of the molecular fluorescence
intensity of a sample of tonic water containing a trace of quinine and, in a general way, some of
the data-domain conversions that are necessary to arrive at a number related to the intensity.
The intensity of the fluorescence is significant in this context because it is proportional to the
concentration of the quinine in the tonic water, which is ultimately the information that we
desire.
The information begins in the solution of tonic water as the concentration of quinine. This
information is teased from the sample by applying to it a stimulus in the form of
electromagnetic energy from the laser shown in Figure 1-3. The radiation interacts with the
quinine molecules in the tonic water to produce fluorescence emission in a region of the
spectrum characteristic of quinine and of a magnitude proportional to its concentration.
Radiation that is unrelated to the concentration of quinine is removed from the beam of light
by an optical filter, as shown in Figure 1-3. The intensity of the fluorescence emission, which is
nonelectrical information, is encoded into an electrical signal by a special type of device called
an input transducer. The particular type of transducer used in this experiment is a
phototransducer. In this example, the input transducer converts the fluorescence from the
tonic water to an electrical current I, proportional to the intensity of the radiation. The
mathematical relationship between the electrical output and the input radiant power impinging
on its surface is called the transfer function of the transducer.

The current from the phototransducer is then passed through a resistor R, which according to
Ohm’s law produces a voltage V that is proportional to I, which is in turn proportional
to the intensity of the fluorescence. Finally, V is measured by the digital voltmeter to provide a
readout proportional to the concentration of the quinine in the sample. Voltmeters,
alphanumeric displays, electric motors, computer screens, and many other devices that serve to
convert data from electrical to nonelectrical domains are called output transducers.

Calibration of Instrumental Methods

A very important part of all analytical procedures is the calibration and standardization process.
Calibration determines the relationship between the analytical response and the analyte
concentration. Usually this is determined by the use of chemical standards.

Almost all analytical methods require some type of calibration with chemical standards.
Gravimetric methods and some coulometric methods are among the few absolute methods

that do not rely on calibration with chemical standards.

An external standard is prepared separately from the sample. By contrast, an internal standard
is added to the sample itself. 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.

However, in some routine analyses, two-point calibrations can be reliable.

Calibration is accomplished by obtaining the response signal (absorbance, emission intensity,

electrode potential, peak area) as a function of the 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 next
step is the prediction step, where the response signal is obtained for the sample and used to
predict the unknown analyte concentration, cx, from the calibration curve or best-fit equation.

The concentration of the analyte in the original bulk sample is then calculated from cx by
applying the appropriate dilution.

factors from the sample preparation steps.

Selecting an Analytical Method

Performance Characteristics of Instruments

Sensitivity
The quantitative definition of sensitivity that is accepted by the International Union of Pure and
Applied Chemistry (IUPAC) is calibration sensitivity, which is the slope of the calibration curve at
the concentration of interest. Most calibration curves that are used in analytical chemistry are
linear and may be represented by the equation:

Detection Limit
The most generally accepted qualitative definition of detection limit is that it is the minimum
concentration or mass of analyte that can be detected at a known confidence level. This limit
depends on the ratio of the magnitude of the analytical signal to the size of the statistical
fluctuations in the blank signal.

Experimentally, Sm can be determined by performing twenty to thirty blank measurements,

preferably over an extended period of time.

Selectivity

Selectivity of an analytical method refers to the degree to which the method is free from
interference by other species contained in the sample matrix. Unfortunately, no analytical
method is totally free from interference from other species, and frequently steps must be taken
to minimize the effects of these interferences.

Dynamic Range
Figure 1-13 illustrates the definition of the dynamic range of an analytical method, which
extends from the lowest concentration at which quantitative measurements can be made (limit
of

quantitation, or LOQ) to the concentration at which the calibration curve departs from linearity
by a specified amount (limit of linearity, or LOL). Usually, a deviation of 5% from linearity is
considered the upper limit. Deviations from linearity are common at high concentrations
because of nonideal detector responses or chemical effects. The lower limit of quantitative
measurements is generally taken to be equal to ten times the standard deviation of repetitive
measurements on a blank, or 10sbl. At this point, the relative standard deviation is about 30%
and decreases rapidly as concentrations become larger.

Example