Professional Documents
Culture Documents
Module Id 02
Pre-requisites
Objectives
Keywords
Analytical chemistry can be divided into areas called qualitative analysis and quantitative
analysis. Qualitative analysis deals with the identification of substances. It is concerned with
what elements or compounds are present in a sample. Quantitative analysis is concerned with
classification of quantitative analysis may be based upon the size of the sample available for
analysis. When a sample weighing more than 0.1 g is available, the analysis is spoken of as
macro; semi-micro analyses are performed on samples of perhaps 10 to 100 mg; micro
analyses deal with samples weighing from 1 to 10 mg; and ultramicro analyses involve
with a suitable reagent which quantitatively precipitates the desired constituent present in the
sample solution. The precipitate which is of known concentration is filtered, washed, dried
and weighed. For example, an excess of dilute sulphuric acid is added to a given solution
containing barium ions. The precipitate of barium sulphate formed is filtered, washed, dried
and weighed. From the weight of barium sulphate the quantity of barium in the given solution
is calculated.
possible sources of error are readily checked, since filtrates can be tested for
completeness of precipitation
it is an absolute method
it is inexpensive.
Another group of techniques was soon developed in which quantitative analysis was achieved
by measuring volume of solutions, hence it was called volumetric. To the sample solution of
unknown concentration, a solution of known concentration is gradually added till the reaction
between them is just complete as shown by some indicator. The volume of the sample and
reagent solutions, the concentration of the reagent solution are also known so the
concentration of the given sample solution can be calculated. For example, a known volume
until the solution in the flask becomes just pink. The volume of sodium hydroxide solution
added is recorded and from this, the concentration of given hydrochloric acid is calculated.
This process is called titration and the determination is termed titrimetric determination.
• Inexpensive
Over the last 50 years or so there has been a growing tendency to make use of certain
the end point in a titration or to follow the course of a chemical reaction. It should be noted
that in such cases the titrimetric methods are not basically altered from their standard
Recently such methods commonly known as the instrumental methods of analysis have been
increasingly used especially in the field of industrial and commercial quantitative analysis
The electroanalytical methods apply an electrical signal to the sample and/or monitor an
electrical property of the sample. The separative methods rely upon separation of the
1. Spectral Methods
If the absorbed energy is electromagnetic radiation in the X-ray, ultraviolet or visible region
A. UV-VIS spectroscopy
A. UV-VIS spectroscopy
electromagnetic radiation. The absorption or reflectance in the visible range directly affects
the perceived colour of the chemicals involved. Ultraviolet and visible radiation absorption
UV-spectrum of particular compound is recorded in conjunction with other spectral data such
B. IR spectroscopy
Infrared Spectroscopy is the analysis of infrared light interacting with a molecule. Molecular
vibrations can occur by two different mechanisms. Quanta of infrared radiation can excite
atoms to vibrate directly. Most organic molecules are fairly large and their resultant
group frequency region, principal absorption bands may be assigned to vibration units
consisting of only two atoms. Such influences reveal themselves on careful study and offer
valuable evidence as to the nature of neighboring atoms. Fingerprint region extends from
1400 cm-1 to 400 cm-1. Absorption bands found here are related to vibrations of molecule as a
Advantages of IR-Spectroscopy:
Non-Destructive
Easy to use
High precision
Inexpensive technique
introduced into a flame. The wavelength of the colour tells us what the element is, and the
In early experiments, visible colour of the flame was used to confirm the presence of certain
elements in the sample, particularly alkali metals and alkaline-earth metals. Later the whole
ultraviolet and visible range was utilized using a spectrophotometer. This instrument
permitted us to select the wavelengths of the radiation and measure its intensity with
considerable accuracy.
The spectrophotometric technique has proven to be one of the most reliable and easily used
magnesium. Flame photometry is also named as flame emission spectroscopy because of the
use of a flame.
High sensitivity and high reliability for determination of elements in first two columns
of periodic table.
Flame photometry is a simple, rapid method for the routine determination of elements
In emission spectroscopy, a sample is excited by absorbing thermal or electric energy and the
radiation emitted by the excited sample is studied for both qualitative and quantitative
analysis. Most of the spectroscopic techniques are related to molecules but emission
metal analysis. With it, all metallic elements can be identified and quantitatively determined
in very low concentrations, as can be metalloids, such as arsenic, silicon and selenium.
present.
The method can be used for quantitative analysis of about seventy elements at
High sensitivity.
2. Electroanalytical Methods
with most of the electroanalytical methods, electrical contact with the sample is completed by
Amperometry is based on the control potential between the two electrodes and the current is
measured.
Potentiometry is based on the controlled current between two electrodes and the potential is
measured.
a current can be applied to the electrodes in the solution. The measurement of quantity Q of
is applied to one of the electrodes while the current flowing through the electrode is
measured. During the current measurement the potential is varied in some predetermined
manner. Polarography is the series of voltammetric methods in which the electrode to which
3. Separative Methods
In the process of separation, the components can be individually assayed either qualitatively
The sample is placed on the edge of the stationary phase (a solid or liquid) and a mobile
phase (a liquid or gas) is admitting to flow over the stationary phase. Strong components
sticks to the stationary phase are swept less rapidly than weakly adhere. Result causes
separation of components
1. liquid chromatography
2. gas chromatography
Depending upon the state or nature of the mobile phase. Some of the chromatographic
Paper chromatography
Thin-layer chromatography
Gas chromatography
A. Paper chromatography
Apparatus
Apparatus required for paper chromatography consists of a support for paper, a solvent
trough, and an air-tight chamber. Size of chamber may vary from an ordinary test-tube to
large aquarium depending on size of paper. Sample is applied prior to dipping into eluting
ultraviolet absorbance,
infrared absorbance,
fluorescence,
radioactivity,
bioautography, or
involves placing paper in contact with culture medium followed by examination of growth of
chromatography method.
Nature of layer: Commonly used adsorbents are silica gel, alumina, diatomaceous earth, and
powdered cellulose. These materials can be combined with binder to make a cohesive layer.
Preparation of layer:
an aqueous slurry of adsorbent over entire surface. Slurry must be neither too thick (viscous)
nor too thin, or it will not spread properly. In commercial spreading machines a slotted trough
travels over glass and deposits a uniform layer. Binder requires about 30 minutes to "set".
The edges should be cleaned and plates stored in a cabinet. Pre-coated TLC plates (glass or
Procedure must be conducted in closed chamber. In order to detect spots, iodine vapor is
used. Another detecting reagent is a spray of sulfuric acid. There are a host of reagents
available, and, of course, spot can be scraped off, eluted, and investigated by any available
method.
Advantages:
C. Gas chromatography
Fig.3.
Carrier gas passes through pressure regulators which control flow rate through apparatus.
Sample is introduced into heated chamber either through silicone rubber septum or by
sampling valve. Carrier gas carries sample components through column where they are
separated. A thermostated oven is provided for column, injector, and detector. There are
Carrier gas
Common carrier gases are helium, nitrogen, hydrogen, and argon. These gases are relatively
inexpensive, and are not hazardous to handle. Choice of carrier gas is usually based on
availability in high purity grade. Thermal conductivity detectors work best with hydrogen or
helium. Helium is popular choice for analysis. Flow through column is caused by difference
in pressure between inlet and outlet. Pressure regulating valves maintain an inlet pressure, Pi
outlet pressure, Po, is normally atmospheric pressure. On some instruments, flow controllers
As gases are compressible, complications arise in determining flow rate and volume of gas
Sample introduction
It is important to introduce sample in shortest time and in smallest volume possible. Sample
chamber may be heated for rapid vaporization of liquid samples. In some instruments, sample
is injected directly into column at inlet. Size of sample is dictated by several factors:
sensitivity of detector
Ordinary chromatograph can handle liquid samples in range of 0.1 to 10 µl and gaseous
Columns
There are two types of columns, packed and open tubular. Packed columns are easier to
fabricate, less expensive, last longer, have higher capacity. Open tubular columns have less
pressure drop. Packed columns are usually 1 to 20 m long. Open tubular columns are usually
10 to 50 m long. Columns are bent in U- or W-shape or coiled to fit oven. Short columns are
often made of glass. Longer columns are made of copper, aluminum, or stainless steel.
Detectors
Separations performed in column must be sensed and recorded. Detector must ignore large
amount of carrier gas and find trace amounts of sample components contained therein.
simple calibration,
low noise,
inexpensive,
robust, and
safe to operate
apparatus and technique, has evolved into a new practice of liquid chromatography which is
competitive with gas chromatography in speed and resolution of complex mixtures. HPLC
Packing Materials:
Various types of materials such as pellicular beads (covered with thin layer of porous
material), silica beads are used for packing the column. Another type of chemically bonded
packing utilizes silicone polymers which are more stable because of their three-dimensional
cross-linked structure.
give a reasonable flow rate. Pressures up to 10,000 psi are not difficult to handle in the small
Detectors:
The smaller columns and faster flow rates place rigid requirements on the detection system.
Flow-through detectors with low dead volumes and high sensitivity are a necessity. Two
types of detectors are currently popular. One is based on a flow-through micro-cell placed in
an ultraviolet spectrophotometer.
difference in refractive index between the pure mobile phase (reference stream) and the
column effluent.
Widespread applicability
Greater reproducibility
High resolution
Electrophoresis is the separative method that takes advantage of the relative mobility of ions
toward an electrode of opposite charge (to the ion) and away from an electrode of similar
charge.
The buffered solution through which the ions normally travel is either supported by porous
paper or is in a gel.
4. Radioanalytical methods
Radioanalytical chemistry focuses on the analysis of sample for their radionuclide content.
Various methods are employed to purify and identify the radio-element of interest through
Good accuracy
They minimize or even eliminate the need for separations that are required in other
analytical methods.
5. Thermoanalytical methods
Thermal analysis is a branch of materials science where the properties of materials are
studied as they change with temperature. Several methods are commonly used:
I. Do you know
1. Analytical chemistry can be divided into areas called qualitative analysis and
quantitative analysis.
element of interest.
molecule. (False)
1. Separations using mass spectrometry are based upon the relative motion in an
ions.
2. The smaller columns and faster flow rates place rigid requirements on the
detection system.
3. Various types of materials such as pellicular beads (covered with thin layer of
porous material), silica beads are used for packing the column.