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Introduction

The document discusses the function and mechanism of a spectrophotometer. A spectrophotometer uses the Beer-Lambert law to determine the concentration of a substance in a solution by measuring how much light the solution absorbs at specific wavelengths. It works by passing light through a sample solution and measuring the intensity of the light transmitted with a photodetector.

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38 views1 page

Introduction

The document discusses the function and mechanism of a spectrophotometer. A spectrophotometer uses the Beer-Lambert law to determine the concentration of a substance in a solution by measuring how much light the solution absorbs at specific wavelengths. It works by passing light through a sample solution and measuring the intensity of the light transmitted with a photodetector.

Uploaded by

Sherlyn cheah
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Introduction

What is the function of spectrophotometer?


Spectrophotometry is a technique in bio chemistry used for quantitative analysis of
substances based on their absorbance of light at specific wavelengths. This instrument
enables researches to quantify the concentration of a particular substance inside a solution
based on its absorbance properties. By passing a beam of light through a sample solution and
measuring the intensity of light transmitted, the spectrophotometer provides valuable data
that can be used to determine the concentration of the sample or analysis.
Beer Lambert Law:
The beer-lambert law, also known as the beer lambert-bouguer law, is a fundamental
principle in spectrophotometry that describes the relationship between the concentration of an
absorbing substance in a solution, the path length of the light through the solution, and the
absorbance of light by the substance (helps us understand how light and substances interact).
It tells us that when light goes through a solution, the more of a substance there is, the more
light gets absorbed.
Mechanism of spectrophotometer:
The mechanism of spectrophotometer involves several key components working
together to measure the absorbance of light by a sample solution. First, a light source emits a
beam of polychromatic( containing multiple wavelengths) or monochromatic(contain only 1
wavelength) light. This light beam then passes through a monochromator, which selects the
desired wavelength of light to be used for the analysis. The selected light then travels through
the sample solution contained in a cuvette or sample cell.
As the light pass through the sample, it interacts with molecules of the
substance/analyte present in the solution. Some of the light is absorbed by the molecules,
while the rest is transmitted through the solution and detected by a photodetector. The photo
detector measures the intensity of the transmitted light, the instrument then converts this
intensity into absorbance values using appropriate calibration and calculation methods. By
comparing the absorbance of the sample of that of standards with known concentrations, the
spectrophotometer allows for accurate determination of the concentration of the substance in
the sample solution.
In simpler sentence, the spectrophotometer works by shining light through a solution.
Some of the light gets absorbed by the substance in the solution, and the rest passes through.
Then, a detector measures how much light makes it through.

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