[go: up one dir, main page]
Scribd
Upload
27 views12 pages

Colorimeter Basics & Applications

The document discusses the principles of colorimetry, emphasizing the Beer-Lambert law which relates the absorbance of light to the concentration of colored solutions. It explains the use of photoelectric colorimeters in clinical chemistry for measuring optical density (OD) and determining substance concentrations through end point and kinetic analysis. Additionally, it highlights the importance of using complementary filters to select the appropriate monochromatic light for accurate measurements.

Uploaded by

aryan8074
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
27 views12 pages

Colorimeter Basics & Applications

The document discusses the principles of colorimetry, emphasizing the Beer-Lambert law which relates the absorbance of light to the concentration of colored solutions. It explains the use of photoelectric colorimeters in clinical chemistry for measuring optical density (OD) and determining substance concentrations through end point and kinetic analysis. Additionally, it highlights the importance of using complementary filters to select the appropriate monochromatic light for accurate measurements.

Uploaded by

aryan8074
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
You are on page 1/ 12

COLORIMETER

Dr. Nisha Jha


Assistant Professor
(Course Co-ordinator Phase-1 MBBS)

Department Of Biochemistry
COLORIMETER

•Colored solutions have the property of absorbing light of


definite wavelengths. The amount of light absorbed or
transmitted by a colored solution is in accordance with the
Beer-Lambert law.

•As per the Beer’s law, the intensity of the color is directly
proportional to the concentration of the colored particles
in the solution.
•The Lambert’s law states that the amount of light absorbed by a
colored solution depends on the length of the column or the depth of
the liquid through which light passes.

•The Beer-Lambert law combines these two laws.


•Photoelectric colorimeter. The components are shown
diagram-matically in left side. The instrument is shown in
the right side.
•The ratio of intensity of emergent light to intensity of
incident light (E / i) is termed as transmittance (T).

•The absorbance is expressed as –log T.

•The Optical Density is calculated as –log T.

•The plot of the concentration versus transmittance is not


linear, but a graph of the concentration against
absorbance (OD) is linear.
Most of the clinical chemistry estimations are done by colorimetric
methods.

A colored derivative of the compound to be measured is prepared


and its OD is measured using a photo electric colorimeter.

This value is compared with that of a standard of known


concentration.
Filter, used for selecting the monochromatic light (mono = single;
chrome = color).

Filters will absorb light of unwanted wavelength and allow only


monochromatic light to pass through. This light will have maximum
absorbance when passed through a particular colored solution.
The color of filter should be complementary to the color of the solution.
Color of filter Wavelength Color of solution
Violet 420 Brown
Blue 470 Yellowish brown
Green 520 Pink
Yellow 580 Purple
Red 680 Green/blue

COLOR OF FILTER AND COLOR OF SOLUTION ARE COMPLEMENTARY


•In clinical laboratory, serum sample and reagents are mixed and
incubated at 37oC for a fixed time, say 10 minutes, to develop the
colour optimally.

•After the incubation period, the OD is ascertained and the


concentration of the substances is calculated.

•This is called end point analysis.


On the other hand, the serum and reagents are
incubated, and readings are taken at 2 and 3 minutes
exactly;
from the difference in OD between the two values, the
concentration is calculated. This is the kinetic analysis.

Here the optimum colour is not developed; but is quicker


and hence is often used in autoanalysers.

You might also like