Principle and application of colorimeter
Introduction:
A colorimeter measures the concentration of colored substances in a solution by analyzing how
much light of a specific wavelength is absorbed or transmitted through the solution. This
measurement is based on the Beer-Lambert Law, which states that absorbance is directly
proportional to the concentration of the colored substance and the path length of the light through
the solution.
Louis J. Duboscq invented it in the year 1870.
Principle:
When an incident light beam with intensity I0 passes through a solution, a part of the incident
light is reflected (Ir) and absorbed (Ia) while the remaining incident light is transmitted (It).
i.e., Io = Ir + Ia + It
The measurement of (I0) in the colorimeter eliminates (Ir), and it is sufficient to calculate the (Ia).
Using cells with the same characteristics maintains a steady amount of light reflection (I r). Then
(I0) & (It) are measured.
The operation of the colorimeter is based on Beer-Lambert’s law which states that the amount of
light absorbed by a color solution is directly proportional to the solution’s concentration and the
length of a light path through it.
A ∝ cl
A = ∈cl
Where,
A = Absorbance/ Optical density of the solution
∈ = Coefficient of absorption
c = Concentration of solution
l = Length of the path
A series of lenses in a colorimeter guide a beam of light with a particular wavelength through a
solution as it makes its way to the measuring apparatus. This compares the colour to a current
standard to examine it. The absorbance or % transmittance is then calculated using a
microprocessor. By measuring the difference between the amount of light at its source and that
after passing the solution, it is possible to determine the concentration of the solution and how
much light will be absorbed.
Beer-Lambert Law:
The core principle is the Beer-Lambert Law, which relates absorbance to concentration and
path length.
Light Interaction:
When light passes through a solution, some light is absorbed by the colored substance, and the
rest is transmitted.
Wavelength Selection:
Colorimeters use a specific wavelength of light that is most strongly absorbed by the substance
being measured.
Absorbance Measurement:
The colorimeter measures the amount of light that is absorbed or transmitted, and this
measurement is used to determine the concentration of the substance.
Quantitative Analysis:
By comparing the absorbance of a sample to a series of standards with known concentrations,
the concentration of the unknown sample can be determined.
Components of a Colorimeter:
Light Source: Provides a beam of light.
Monochromator: Selects a specific wavelength of light.
Cuvette Holder: Holds the sample cuvette.
Detector: Measures the intensity of the transmitted light.
Readout Device: Displays the absorbance or transmittance value.
Applications:
Biochemistry: Determining the concentration of biomolecules in biological samples like blood
and urine.
Environmental Science: Measuring pollutants in water and soil.
Food and Beverage Industry: Analyzing the color and quality of food and drinks.
Pharmaceuticals: Quality control of medications and drug analysis.
Textile and Paint Industries: Color matching and quality control.
Cosmetics: Evaluating UV protection of skincare products.
Medical Diagnostics: Assessing hemoglobin levels in blood, among other tests.
Industrial Quality Control: Monitoring color consistency in various manufacturing processes.
Applications of Colorimeter
Colorimeters are widely used in biochemistry for various applications, including measuring glucose
levels in blood, protein concentrations, and enzyme activity.
Blood samples are tested using a colorimeter to determine the amount of hemoglobin present.
Advantages of Colorimeter
A quick and affordable means of evaluating quality is the colorimeter.
The Colorimeter makes it simple to perform a quantitative examination of colored chemicals.
Results are available in under a second.
Four AA batteries can be measured using a portable colorimeter between 100 and 300 times.
Disadvantages of Colorimeter
The procedure of determining the concentration of colorless substances becomes laborious.
Colorimeter does not function in the ultraviolet or infrared spectrum since it only measures
wavelength absorbance in the visible range of light (400nm to 700nm).
A spectrum range must be set rather than a specific wavelength to measure the absorbance.
Measurements might be challenging on surfaces that reflect light.
Principle and application of Spectrophotometer
Introduction:
A spectrophotometer measures the absorbance or transmittance of light through a sample at
specific wavelengths, and this measurement is used to determine the concentration of a substance
or analyze its properties. It works by passing a beam of light through the sample and then
detecting the amount of light that is transmitted or absorbed. The data obtained can be used to
identify and quantify substances, analyze chemical kinetics, and study the structure of
molecules.
Principle:
The core principle of a spectrophotometer is based on Beer-Lambert Law, which states that the
absorbance of a solution is directly proportional to the concentration of the absorbing substance
and the path length of the light beam through the solution.
Light Source:
Emits a beam of light, which can be in the ultraviolet (UV) or visible (Vis) range, or both.
Monochromator:
This component selects a specific wavelength of light from the source, ensuring
monochromatic light (light of a single wavelength) is used.
Sample Holder:
A cuvette (specialized test tube) holds the sample solution, and the light beam passes through
it.
Detector:
Measures the intensity of the light that passes through the sample.
Readout System:
Displays the absorbance or transmittance values, often as a graph or numerical data.
Applications:
Spectrophotometers are used in a wide range of fields due to their ability to analyze the
interaction of light with matter:
Chemistry: Determining the concentration of solutions, identifying unknown compounds, and
studying chemical reactions.
Biochemistry: Analyzing enzyme kinetics, protein concentrations, and DNA/RNA
quantification.
Biology: Monitoring dissolved oxygen in aquatic ecosystems, studying microbial growth, and
analyzing cellular processes.
Environmental Science: Detecting pollutants in water and air.
Quality Control: Ensuring color consistency in manufacturing processes, particularly in
industries like printing, textiles, and food production.
Forensics: Analyzing evidence in criminal investigations.
Astronomy: Measuring the spectra of celestial objects to understand their composition and
properties.
In essence, spectrophotometers provide a powerful tool for analyzing the properties of materials
by measuring their interaction with light across different wavelengths.
Applications
Some of the major applications of spectrophotometers include the following:
Detection of concentration of substances
Detection of impurities
Structure elucidation of organic compounds
Monitoring dissolved oxygen content in freshwater and marine ecosystems
Characterization of proteins
Detection of functional groups
Respiratory gas analysis in hospitals
Molecular weight determination of compounds
The visible and UV spectrophotometer may be used to identify classes of compounds in both
the pure state and in biological preparations.
