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Home / Chemistry / Understanding the Principle of Spectrophotometers - Testbook

Understanding the Principle of Spectrophotometers - Testbook

Last Updated on Feb 18, 2025
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A spectrophotometer is a device that measures how much light a sample absorbs. This is achieved by directing a light beam through a sample and then measuring the intensity of the light on the other side. These devices are crucial in maintaining colour accuracy throughout the production process and are widely used by manufacturers and researchers. The versatility of spectrophotometers means they can be used in nearly every commercial and industrial sector. Their primary applications include analyzing liquids, plastics, paper, metals, and fabrics to ensure the consistency of the chosen colour from conception to the final product.

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Spectrophotometer Instrumentation

A spectrophotometer consists of two main parts: a spectrometer, which produces light of any wavelength, and a photometer, which measures the intensity of light. The design of the spectrophotometer places the sample between the spectrometer and the photometer. As the light passes through the sample, the photometer measures the intensity of the light and sends a voltage signal to the display. Any changes in the absorption of light will affect this voltage signal. Spectrophotometers come in various shapes and sizes, each designed for specific applications and functionalities. The most commonly used types include 45 degrees, sphere, and multi-angle spectrophotometers. Another concept closely related to spectrophotometry is Spectroscopy , which measures the absorption and intensity of light from its source.

The basic components of a spectrophotometer include a light source, a digital display, a monochromator, a wavelength sector for transmitting a selected wavelength , a collimator for straight light beam transmission, a photoelectric detector, and a cuvette for holding the sample.

The symbol l 0 represents the intensity of light, which measures the number of photons per second. When light passes through a blank solution that does not absorb light, it is symbolized as (l). Absorbance (A) and Transmittance (T) are other important factors in spectrophotometry.

 

 

 

 

To measure the intensity of light that passes a blank solution and later measure the intensity of light passing a sample. Then calculate the transmittance and the absorbance. For the measurement of absorbance, we can use an isosbestic point where the absorbance and wavelength of two or more species are the same.

The number of protons that transmit and absorb is dependent on the length of the cuvette and the concentration of the sample.

The relationship between transmittance and absorption is:

 

 

The transmittance of an unknown sample can be calculated using the formula given below.

 

 

Here,

 

There are several types of spectrophotometer devices such as UV Spectrometry, atomic emission spectrophotometry, and atomic absorption spectrophotometry. They can also be classified based on the range of light source wavelengths, such as IR spectrophotometer and UV-visible spectrophotometer. Spectrophotometry has many practical applications in various fields, including the production of laundry soap, carpeting, and small items like toys or intricate machinery. The two main types of spectrophotometers are portable and bench spectrophotometers, each with its unique uses.

 

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Frequently Asked Questions

A spectrophotometer is an analytical instrument used for the objective calculation of visible light, UV light, or infrared light emission or reflection. Spectrophotometers measure intensity as a function of the wavelength of the light source.

Spectrophotometry is a procedure for determining how much light is reflected by a chemical material by measuring the strength of light as a light beam travels through the sample solution. The fundamental theory is that light is absorbed or emitted over a certain wavelength spectrum by each compound.

A spectrometer is an aspect of the most responsible spectrophotometer for the calculation of different objects. A spectrophotometer is a comprehensive device that involves a light source, a way of collecting the light that has interacted with the objects being measured, and a measurement spectrometer.

The source of light is given by a lamp. The light beam strikes the diffraction grating, which acts like a mirror and divides the light into the wavelengths of its elements. The grating is rotated such that the exit slit is only penetrated by a single wavelength of light. Then with the sample, the light interacts.

A blank is a sample containing everything except for the significance analyte. For example, if you are conducting an experiment with UV-Vis to measure Green Fluorescent Protein concentrations, the protein must be dissolved in a solvent. The blank is a sample of the solvent itself.
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