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Asad Khalil CLE 2

A spectrophotometer is a device used to measure the amount of light absorbed by a sample at different wavelengths, consisting of a light source, sample holder, diffraction grating or prism, and a detector. It is commonly used in chemistry labs to analyze the properties of light and matter, allowing for applications such as determining unknown concentrations and identifying materials. The document also explains the working principle of spectrophotometers, their common types, and the differences between spectrometers and spectrophotometers.

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14 views6 pages

Asad Khalil CLE 2

A spectrophotometer is a device used to measure the amount of light absorbed by a sample at different wavelengths, consisting of a light source, sample holder, diffraction grating or prism, and a detector. It is commonly used in chemistry labs to analyze the properties of light and matter, allowing for applications such as determining unknown concentrations and identifying materials. The document also explains the working principle of spectrophotometers, their common types, and the differences between spectrometers and spectrophotometers.

Uploaded by

asadrajput43211
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© © All Rights Reserved
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Assignment 2

NAME : ASAD KHALIL

CLASS : BMET SEM 6TH

ROLL NO : 70098507

TOPIC : Spectrophotometers

SUBMITTED BY: ASAD KHALIL

SUBMITTED TO: SIR AZIZ


What Is a Spectrophotometer:
A spectrophotometer is a device that is used to measure
the amount of light absorbed by a sample at different wavelengths. A spectrophotometer
measures the amount of light that can pass through a sample. It consists of a light source, a
sample holder, a diffraction grating or prism to separate the light into its different wavelengths,
and a detector to measure the amount of light absorbed at each wavelength.

Spectrophotometry is a standard technique in many chemistry labs. From the data collected, a
computer plots an absorbance spectrum for the sample. Spectrophotometry is the action of using
a spectrometer to take a measurement.

In simple terms, a spectrophotometer is a tool that helps scientists and researchers study the
properties of light and how it interacts with different materials. By measuring the amount of light
absorbed by a sample at different wavelengths, a spectrophotometer can provide valuable
information about the composition and properties of the sample.

A spectrophotometer can be a useful tool for conducting experiments and studying the properties
of light and matter. For example, a student could use a spectrophotometer to study the absorption
spectrum of a solution, which is the pattern of light absorption at different wavelengths. By
analyzing the absorption spectrum of a solution, a student can learn about the composition and
properties of the solution.

The basic idea of spectrophotometry is that light passes through a sample and the intensity of the
beam is compared before and after the sample. Different samples will absorb light differently
and allow different amounts to pass-through through different colors of light.

This technique works because each molecule absorbs light. Depending on the molecules it will
absorb certain colors more than others.

As an example, spectrometry at visible wavelengths is the easiest kind to visualize. A black


sample will absorb generally absorb all colors of visible light. That is the reason it appears black.
On the other side, if all wavelengths of visible light pass through the sample then the sample
most likely appears white.
How does a spectrophotometer work?
The simplest spectrometer includes a light source, a sample holder, and a detector.

The light source produces the photons that will pass through the sample. The exact type of light
source will depend on the wavelength of light needed. Depending on the source, a collimator and
prism select the correct wavelength.

The light interacts with the sample next. A quartz cuvette holds the samples. A cuvette is a
specialized piece of glassware with a very precise width and material. The material of the sample
holder is important. You don’t want to use a sample holder that will also absorb the wavelength
of light you are investigating. This is also why it is best practice to run a blank/background in the
instrument with an empty cuvette. The path length through the cuvette is also an important
parameter for some calculations. The path length is the amount of sample the light passes
through. So a cuvette that is 1 cm wide has a path length of 1 cm.

After the light passes through the sample, it travels to the detector. Similar to the light source, the
exact detector will depend on the wavelength of light. Some of the most common detectors are
photomultiplier tubes and photodiodes. The detector counts the number of photons reaching it.
The detector connects to a computer that plots this data. This plot is called an absorption
spectrum. It is a plot of light intensity versus the wavelength of light. An example of what an
absorption spectrum looks like is seen below.
Most Common Types of Spectrophotometers :
Spectrometers are generally classified based on the wavelength of light the source is. There are
two main classifications:

UV-Vis Spectrophotometer: The UV-Vis spectrophotometer is commonly referred to as just


‘UV-Vis’. This instrument measures absorbance in the ultraviolet and visible range. Generally,
that means 200 nm – 700 nm.

IR Spectrophotometer: The IR spectrophotometer measures samples using infrared light, 700-


15000 nm.

Applications of Absorption Spectra


Absorption spectra have a lot of different possible uses:

Determining an Unknown Concentration: Absorption spectra can be used to determine the


concentration of a solution of unknown concentration. Absorption data of a series of solutions of
known concentrations creates an absorbance curve. Then, the unknown solution can be compared
to this curve to determine the concentration. This is also known as Beer-Lambert
Law.Identifying a Material: Each pure material will have a unique absorption spectrum
depending on the structure of the molecule. Therefore, an absorption spectrum helps to identify
an unknown.Identifying Functional Groups Present: Some functional groups will have distinct
spectral signatures. Identifying these in a spectrum can inform if that functional group is present
and to test if a reaction was successful.Common labs with spectrophotometers include chemistry,
biochemistry, physics, clinical, and materials labs.
What is spectrophotometer used for:

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.

What is the basic principle of spectrophotometer:

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.

What is the difference between a spectrometer and a spectrophotometer:

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.

How does a spectrophotometer work:

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.

What is a blank in spectrophotometry:

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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