Spectrophotometer

A spectrophotometer is an instrument that measures the amount of light absorbed by a substance. Every substance transmits and absorbs light by different amounts. Knowing how much this light gets absorbed gives us an identity and helps quantify different materials. Thus it is a quantitative measurement of properties of a material as a function of wavelength.

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Detailed Description for Spectrophotometer

Spectrophotometer uses photometers as a means to measure light intensity as a function of its wavelength. The most vital feature of spectrophotometers is its spectral bandwidth, percentage of sample transmission, and log of range of sample absorption and reflectance measurement. 
There are two types of devices,
1.    Single beam photometry: It measures the relative intensity of light before and after inserting a test sample. Example: micro
2.    Double beam photometry: It measures the absorption of wavelength by comparing the light intensity between two optical paths.
PARTS
1.    Sample solution is placed inside the spectrophotometer.
2.    A light source is directed towards the sample
3.    A device called monochromator splits the light into its constituent colors. Each colour has its own wavelength.
4.    An adjustable slit allows only one wavelength to pass through the sample.
5.    The sample solution is held in a container called cuvette. Wavelength of the light hits this container.
6.    The wavelength of light coming out through the solution is read on the output screen. This determines how much of the light is absorbed by the sample.
WORKING
The instrument consists of a lamp which acts as a source of light. The light beam coming from the lamp is made to strike a diffraction grating placed between the source and the sample. The grating acts like a prism and separates the light into its constituent wavelength. The grating is turned to a desired angle to make only a specific wavelength to reach the slit. Finally, the light interacts with the sample. The detector is made to measure the amount of light that passes through the sample completely. When this light reaches the detector the amount of light absorbed will also be determined. The measurements are converted and displayed on a digital read out.
 The detector measures the transmittance and absorbance of the s
USES
•    This is a reliable method of determining the composition of a substance since every chemical compound absorbs different wavelengths to different amounts.
•    It is used in all areas of scientific research like, microbiology, biochemistry, forensics, physics and medicine.
•    To measure the ingredients present in a drug to make sure it’s safe for intake.
•    To measure bacterial growth and diagnosis of a patient
•    Scientists use it to study the progress of a chemical reaction. It will also help to find short lived products in a reaction.
APPLICATIONS
1.    To estimate dissolved organic carbon concentration
2.    Aromaticity is measured in terms of specific UV absorption
3.    It is a major part of Bial’s test to determine concentration of pentose.
4.    It is an important technique in biochemical experiments that involve biochemical analysis of DNA, RNA, protein isolation etc.
5.    UV spectrophotometer is used to separate mixture of compounds in analytical chemistry and to purify individual components.
6.    In enzyme assay, where the appearance and disappearance of light absorbing materials can be measured both quantitatively and qualitatively.
7.    To determine the molecular weight of a derivative compound.

 

 

 

 

 

 

 

 

 


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