1.

Colorimetric Methods
Colorimetric analysis is a method of determining the concentration of a chemical element or chemical
compound in a solution with the aid of a color reagent. It is applicable to both organic compounds and
inorganic compounds and may be used with or without an enzymatic stage. The method is widely used in
medical laboratories and for industrial purposes, e.g. the analysis of water samples in connection with
industrial water treatment.
The equipment required is a colorimeter, some cuvettes and a suitable color reagent. The process may
be automated, e.g. by the use of an AutoAnalyzer or by Flow injection analysis. Recently, colorimetric
analyses developed for colorimeters have been adapted for use with plate readers to speed up analysis
and reduce the waste stream.

2.

Volumetric Methods

Titrimetry refers to that group of analytical techniques which takes advantage of titers or concentrations of
solutions. In metallurgy the word "titer" refers to the fineness of gold or silver. In chemistry it is the
measure of the concentration of a solution. In medicine it is defined as the extent to which an antibody
solution can be diluted before it ceases to give a positive reaction with an antigen.
Though in chemistry the term titrimetry often refers to the use of some volume of a solution of known
concentration to determine the quantity of analyte, there are still some variations on the use of the term. It
is used rather to denote a quantity of some other measurement parameter which relates directly to the
quantity of analyte which is to be measured:

Volumetric titrimetry establishes a quantity of analyte using volumes of reagents of known

concentrations and the knowledge of the stoichiometry of the reactions between the reagents and
the analyte(s).
Gravimetric titrimetry determines the quantity of analyte by a measure of the mass of a solution of
known concentration.
Coulometric titrimetry arrives at the amount of analyte by measuring the duration of a given
electrical current. Since amperes x time = coulombs or total charge, the number of equivalents of
analyte can be measured by relating the extent of reaction to the number of moles of electrons
(Faradays).

The equivalence point is the point at which a volume, a mass or a quantity of charge equivalent to the
amount of analyte present in the sample to be measured is reached. It is the point of stoichiometric
chemical equivalence.
The end point is the point at which some detection technique tells you that chemical equivalence has
been reached. The end point may occur before or after the equivalence point, giving a titration error. It is
for this reason that blank samples are often used. Blank samples are prepared so that you have a
measure of the amount that needs always to be added to or subtracted from the end point (the titration
error) to achieve the equivalence point.

3. Turbidimetry
Turbidimetry (the name being derived from turbidity) is the process of measuring the loss of intensity of
transmitted light due to the scattering effect of particles suspended in it. Light is passed through a filter
creating a light of known wavelength which is then passed through a cuvette containing a solution. A
photoelectric cell collects the light which passes through the cuvette. A measurement is then given for the
amount of absorbed lightTurbidimetry
4. Nephelometry
Nephelometry is a technique used to determine the levels of several blood plasma proteins. It is
important in quantification of free light chains in diseases such as multiple myeloma. Quantification is
important for disease classification and for disease monitoring once a patient has been treated (increased
skewing of the ratio between kappa and lambda light chains after a patient has been treated is an
indication of disease recurrence).
It is performed by measuring the turbidity in a water sample by passing light through the sample being
measured. In nephelometry the measurement is made by measuring the light passed through a sample at
an angle.
This technique is widely used in clinical laboratories because it is relatively easily automated. It is based
on the principle that a dilute suspension of small particles will scatter light (usually a laser) passed
through it rather than simply absorbing it. The amount of scatter is determined by collecting the light at an
angle (usually at 30 and 90 degrees).
5. Electrophoresis
Electrophoresis is the motion of dispersed particles relative to a fluid under the influence of a spatially
uniform electric field. This electrokinetic phenomenon was observed for the first time in 1807 by
Ferdinand Frederic Reuss (Moscow State University), who noticed that the application of a constant
electric field caused clay particles dispersed inwater to migrate. It is ultimately caused by the presence of
a charged interface between the particle surface and the surrounding fluid. It is the basis for a number of
analytical techniques used in biochemistry for separating molecules by size, charge, or binding affinity.
Electrophoresis is a technique used in laboratories in order to separate macromolecules based on size.
The technique applies a negative charge so proteins move towards a positive charge. This is used for
both DNA and RNA analysis. Polyacrylamide gel electrophoresis (PAGE) has a clearer resolution than
agarose and is more suitable for quantitative analysis. In this technique DNA foot-printing can identify how
proteins bind to DNA. It can be used to separate proteins by size, density and purity. It can also be used
for plasmid analysis, which develops our understanding of bacteria becoming resistant to antibiotics

6. Chromatography
Chromatography is the collective term for a set of laboratory techniques for the separation of mixtures.
The mixture is dissolved in a fluid called the mobile phase, which carries it through a structure holding
another material called the stationary phase. The various constituents of the mixture travel at different
speeds, causing them to separate. The separation is based on differential partitioning between the mobile
and stationary phases. Subtle differences in a compound's partition coefficient result in differential
retention on the stationary phase and thus changing the separation.

Chromatography may be preparative or analytical. The purpose of preparative chromatography is to

separate the components of a mixture for more advanced use (and is thus a form of purification).
Analytical chromatography is done normally with smaller amounts of material and is for measuring the
relative proportions of analytes in a mixture. The two are not mutually exclusive.

7. Fluorometry
The measurement of fluorescence emitted by compounds when exposed to ultraviolet or other intense
radiant energy. The atoms of certain substances produce fluorescence of a characteristic color and
wavelength, allowing identification and quantification of several clinically significant compounds in biologic
specimens. Although fluorometry is a highly sensitive method of analysis, test interference by other
compounds, especially drugs, may limit its usefulness in some situations.
A fluorometer or fluorometer is a device used to measure parameters of fluorescence: its intensity and
wavelength distribution of emission spectrum after excitation by a certain spectrum of light. These
parameters are used to identify the presence and the amount of specific molecules in a medium. Modern
fluorometers are capable of detecting fluorescent molecule concentrations as low as 1 part per trillion.
Fluorescence analysis can be orders of magnitude more sensitive than other techniques. Applications
includechemistry/biochemistry, medicine, environmental monitoring. For instance, they are used to
measure chlorophyll fluorescence to investigate plant physiology.

8. Chemiluminescence
Chemiluminescence is the emission of light (luminescence), as the result of a chemical reaction. There
may also be limited emission of heat. Given reactants A and B, with an excited intermediate ,
[A] + [B] [] [Products] + light
For example, if [A] is luminol and [B] is hydrogen peroxide in the presence of a suitable catalyst we have:
luminol + hydrogen peroxide 3-APA[] 3-APA + light
where:

3-APA is 3-aminophthalate
3-APA[] is the vibronic excited state fluorescing as it decays to a lower energy level.
The decay of this excited state[] to a lower energy level causes light emission. In theory, one photon of
light should be given off for each molecule of reactant. This is equivalent to Avogadro's number of
photons per mole of reactant. In actual practice, non-enzymatic reactions seldom exceed 1% Q C,
quantum efficiency.
In a chemical reaction, reactants collide to form a transition state, the enthalpic maximum in a reaction
coordinate diagram, which proceeds to the product. Normally, reactants form products of lesser chemical
energy. The difference in energy between reactants and products, represented as

, is turned

into heat, physically realized as excitations in the vibrational state of the normal modes of the product.
Since vibrational energy is generally much greater than the thermal agitation, it rapidly disperses in the
solvent through molecular rotation. This is how exothermic reactions make their solutions hotter. In a
chemiluminescent reaction, the direct product of a reaction is an excited electronic state, which then
decays into an electronic ground state through either fluorescence or phosphorescence, depending partly
on thespin state of the electronic excited state formed.
Chemiluminescence differs from fluorescence in that the electronic excited state is derived from the
product of a chemical reaction rather than the more typical way of creating electronic excited states,
namely absorption. It is the antithesis of a photochemical reaction, in which light is used to drive an
endothermic chemical reaction. Here, light is generated from a chemically exothermic reaction.

9. Osmometry
Osmometry is a laboratory technique to find the molecular weight of an unknown compound by
determining the osmotic strength of a solution.
Osmometers can find the osmotic strength of a solution or colloid using data from a semipermeable
membrane, freezing point depression or vapor pressure.
An osmometer is a device for measuring the osmotic strength of a solution, colloid, or compound.
There are several different techniques employed in osmometry:

Vapor pressure depression osmometers determine the concentration of osmotically active particles that
reduce the vapor pressure of a solution.
Membrane osmometers measure the osmotic pressure of a solution separated from pure solvent by a
semipermeable membrane.
Freezing point depression osmometer may also be used to determine the osmotic strength of a solution,
as osmotically active compounds depress the freezing point of a solution.

10. Electrochemistry Techniques

Electrochemistry is the branch of physical chemistry that studies chemical reactions which take place at
the interface of an electrode, usually a solid metal or a semiconductor, and an ionic conductor, the
electrolyte. These reactions involve electric charges moving between the electrodes and the electrolyte
(or ionic species in a solution). Thus electrochemistry deals with the interaction between electrical energy
and chemical change.
When a chemical reaction is caused by an externally supplied current, as in electrolysis, or if an electric
current is produced by a spontaneous chemical reaction as in a battery, it is called an electrochemical
reaction. Chemical reactions where electrons are transferred directly between molecules and/or atoms
are called oxidation-reduction or (redox) reactions. In general, electrochemistry describes the overall
reactions when individual redox reactions are separate but connected by an external electric circuit and
an intervening electrolyte.

Oxidation and Reduction - The term "redox" stands for reduction-oxidation. It refers to
electrochemical processes involving electron transfer to or from a molecule or ion changing its
oxidation state. This reaction can occur through the application of an external voltage or through
the release of chemical energy. Oxidation and reduction describe the change of oxidation state
that takes place in the atoms, ions or molecules involved in an electrochemical reaction. Formally,
oxidation state is the hypothetical charge that an atom would have if all bonds to atoms of
different elements were 100% ionic. An atom or ion that gives up an electron to another atom or
ion has its oxidation state increase, and the recipient of the negatively charged electron has its
oxidation state decrease.

Electrolysis - The spontaneous redox reactions of a conventional battery produce electricity

through the different chemical potentials of the cathode and anode in the electrolyte. However,
electrolysis requires an external source of electrical energy to induce a chemical reaction, and
this process takes place in a compartment called an electrolytic cell.

Corrosion - Corrosion is the term applied to steel rust caused by an electrochemical process.
Most people are likely familiar with the corrosion of iron, in the form of reddish rust. Other
examples include the black tarnish on silver, and red or green corrosion that may appear on
copper and its alloys, such as brass. The cost of replacing metals lost to corrosion is in the multibillions of dollars per year.