the ppt is related to ife science and spectrphotometer

1. Applications of
Spectrophotometer in
Life Science
Dr. Rahil Razak. Ph.D.

2. Contents of Presentation
Biophysics is an interdisciplinary science that applies approaches and methods traditionally used in
physics to study biological phenomena.
Introduction of Spectroscopy Sectroscopy is a branch of light physics
Historical Background "Spectroscopy, a beacon in the scientific voyage of discovery, unveils the history of
elements, revealing the cosmic symphony written in the light of stars."
Principle of Spectrophotometer "Spectroscopy unveils the symphony of light, decoding the principles that paint the
vibrant portrait of molecular mysteries."
Types of Spectroscopy "In the symphony of science, spectroscopy unveils the unique melodies of molecules
through the harmonious dance of light and matter."
Applications in different Fields Spectroscopy, the visionary lens unlocking a myriad of applications, from probing the
mysteries of distant galaxies to unravelling the intricate tapestry of molecules in the palm
of scientific exploration."
Application in Life sciences "In the delicate tapestry of life science, spectroscopy threads the needle, unravelling the
secrets of biomolecules and illuminating the pathways to revolutionary advancements in
medicine, diagnostics, and biotechnology.
Sensitivity and Precision of
Spectroscopy
"Spectroscopy tests epitomize precision with their dual virtues—sensitivity, finely attuned
to detect subtle signals, and specificity, a beacon ensuring accurate identification,
collectively unraveling the nuanced intricacies of molecular landscapes."

3. Difinitions we should know
01
04
02
05
03
06
Monochromatic Light Absorbance Transmitance
Spectrum solution Chromophores

4. Introduction
Spectrophotometry:
● Spectrophotometry is a quantitative
analytical technique that involves measuring
the intensity of light absorbed or transmitted
by a substance as a function of its
wavelength. This method is widely used in
chemistry, biochemistry, and other scientific
disciplines to determine the concentration of
a substance in a sample, study the absorption
and emission characteristics of molecules,
and provide valuable insights into the
composition and properties of materials
based on their interaction with light
● Spectrophotometer: An instrument for
measuring the intensity of light of a definite
wavelength transmitted by a substance or a
solution.
Nobel laureate Bruce Merrifield
referred to the UV-Vis
spectrophotometer as “probably
the most important instrument ever
developed toward the
advancement of bioscience.”
Visible
Spectrophotometer
Visible
Spectrophotometer

5. Components of spectrophotometer
A spectrophotometer
measures how light
interacts with atoms or
molecules in a sample.

6. Brief History of Spectroscopy
● In the historical development of spectroscopy, following the
fundamental studies of crude spectra of sunlight by Isaac Newton in
1672, certain contributions and achievements are especially
noteworthy.
● The significance of using a narrow slit instead of a pinhole or round
aperture to produce spectra lines, each one an image of the slit and
representing a different color or wavelength, was demonstrated
independently by W. H. Wollaston in 1802 and by Joseph Fraunhofer
in 1814.
● Fraunhofer made many subsequent contributions to optics and
spectroscopy, including first observation of stellar spectra, discovery
and construction of transmission diffraction gratings, first accurate
measurements of wavelengths of the dark lines in the solar spectrum,
and invention of the achromatic telescope.
● The origin of the dark Fraunhofer lines in the solar spectrum was
accounted for by G. R. Kirchhoff in 1859 on the basis of absorption by
the elements in the cooler Sun's atmosphere of the continuous
spectrum emitted by the hotter interior of the Sun.

7. Historical Background of Spectrophotometer
In the 1930s, vitamin research indicated that several vitamins, particularly vitamin A, absorb
ultraviolet (UV) light. Spurred by the American government’s interest in measuring vitamin content
in soldiers rations using ultraviolet and visible (UV-Vis) light, this research culminated in the
commercial launch of UV-Vis spectrophotometers in the early 1940s. Of these, the Beckman DU
spectrophotometer—first sold in 1941—distinguished itself from competing products by delivering
more accurate results and reducing analysis time from hours, or even weeks, to minutes

8. Historical Background of Spectrophotometer
In 1935, Arnold O.
Beckman
founds National
Technologies
Laboratories—
later named
Beckman
Instruments.
In 1941, Beckman introduces the
DU UV-Vis
spectrophotometer, which
has higher resolution and
lower stray light in the
ultraviolet region than any
other commercial instrument.
In 1946, Cary Instruments is
founded by Howard Cary,
George W. Downs, and
William C. Miller under the
name Applied Physics
Corporation. Previously,
Howard Cary was vice
president in charge of
development for Beckman
Instruments.
In 1947, Applied Physics
Corporation delivers the first
commercially available
recording UV-Vis
spectrophotometer, the Cary
11, to the Mellon Institute in
Pittsburgh, PA.
1950s – 1970s Mass production reduces the cost of
UV-Vis spectrophotometers. New photodiode
arrays collect all wavelengths simultaneously,
reducing the time required to scan a spectrum
from minutes to seconds. In 1950, National
Technologies Laboratories changes its name to
Beckman Instruments, Inc.
In 1953, Bausch & Lomb introduces the
SPECTRONIC 20 UV-Vis spectrophotometer,
the first mass produced, low-cost UV-Vis
spectrophotometer.
In 1954, Applied Physics Corporation launches the
Cary 14 spectrophotometer, the first
commercially available double-beam
spectrophotometer. The double beam design
greatly simplifies and speeds up sample
analysis by simultaneously measuring sample
and solvent transmittance over the wide
spectral range of ultraviolet, visible, and near
infrared wavelengths.
1930s 1940s 1950s

9. Historical Background of Spectrophotometer
In 1963, JASCO introduces the
ORD/UV-5 with double-
beam UV-Vis capabilities.
In 1966, Applied Physics
Corporation is purchased
by Varian Medical
Systems, becoming the
Cary Instruments division
of Varian.
In 1969, Cecil Instruments
introduces the CE 212, the
world's first commercially
available variable
wavelength detector for
HPLC, allowing users to
select—without changing
filters or lamps—
detection wavelengths on
a single detector.
In 1979, Hewlett-Packard
launches the first
commercially available
diode-array
spectrophotometer, the
8450A. Unlike
traditional scanning
spectrophotometers
with a single
photomultiplier tube
that scans one
wavelength at a time,
the 8450A utilizes an
array of photodiodes to
scan simultaneously
the full spectrum of
wavelengths in
seconds.
The proliferation of personal computers in
the 1980s improves data acquisition and
instrument control. In 1980, Bausch & Lomb
introduces the Spectronic 2000 UV-Vis
spectrophotometer, the first microprocessor-
controlled double-beam UV-Vis
spectrophotometer. Now, instead of measuring
sample and solvent transmittance separately,
which the single-beam spectrophotometers
required, the double-beam design greatly
simplifies and speeds up sample analysis by
simultaneously measuring sample and solvent
transmittance.
In 1987, Pye Unicam Corporation. introduces the PU-
8700 UV-Vis spectrophotometer, the first
mouse-driven, graphical interface UV-Visible
spectrophotometer.
In 1989, Dr. Arnold O. Beckman, now 88 years old,
receives the National Medal of Science for his
leadership in analytical instrumentation
development.
1960s 1970s 1980s

10. Historical Background of Spectrophotometer
.
In 2010, Agilent Technologies acquires
Varian Inc. and continues to offer
the Cary spectrophotometer series
under the Cary name.
Also in 2010, Thermo Scientific
introduces the Evolution 200 Series
spectrophotometer. Its application-
focused beam geometry tailors the
instrument's optical system to
specific applications for microcells,
solid sampling, and fiber optics.
In 2011, Agilent Technologies
releases the Cary 60 UV-Vis
spectrophotometer with low cost
of ownership—the xenon lap
typically lasts 10 years—and
remote sampling options that
minimize sample handling.

11. Light consists of electromagnetic radiation with a wide
range of frequencies, wavelengths, and energies that all
travel at the same speed. This includes the visible
spectrum and radiation of longer and shorter
wavelengths on either side of the band
Principle
The spectrophotometer principle depends upon the Beer-
Lambert law, which states that a beam of light incidents on
the homogenous solution, reflects some fraction of incident
light, absorbs some light and transmits the remaining light
through the solution.
Both Beer and Lambert have given their own theories on the
absorption of radiation. According to Beer-Lambert law, the
absorbance is equal to the given mathematical expression
A= log I0/I
A= ε C l
Where, A= Absorbance of light
I0= Intensity of incident light
I= Intensity of transmitted light
ε= Absorption coefficient
C= Concentration of the absorbing material
l= Path length (cm)

12. The amount of light transmitted from the solution is inversely
proportional to the absorption of light.
A= log10 1/T
As the absorbance of the medium increases, the transmittance of
light through the solution decreases and vice versa. Absorbance is
a non-dimensional quantity whose value ranges between 0-1.
Transmittance

13. Classes of Spectrophotometer
• In this type, before
reaches the sample the
light source is split into
two separate beams.
From these one passes
through the sample and
second one is used for
reference. This gives the
advantageous because at
the same time the
reference reading and
sample reading can take
place.
• In this type, all the
light passes
through the sample.
To measure the
intensity of the
incident light the
sample must be
removed so that all
the light can pass
through. This type
of spectrometer is
usually less
expensive and less
complicated.
Single Beam Double Beam

15. Applications of spectrophotometry in
Biological sciences
• A spectrophotometer helps in the qualitative analysis of
the properties like type, molecular weight and structure
of different compounds, as the different compounds
absorb light at different wavelengths. Aliphatic or acyclic
hydrocarbons or their derivatives absorb light at a
wavelength ranging between 220-280 nm.
• It also helps in quantitative analysis of proteins, Nucleic
acids, enzymes, amino acid (tyrosine), and blood glucose
level by using a UV-visible spectrophotometer.

16. Applications of spectrophotometry in Biological sciences
● Scientists have a magnitude of ways to determine the
composition of various biological substances. One of
those ways is through the use of spectrophotometry.
● Spectrophotometry has come a long way to
incorporate computers to determining the various
settings that are required to monitor the biological
matter (Baker, 2010).
● Everything from the signaling the pathways in which
proteins act, to the behavior of nucleic acids are
explored during the application of spectrophotometry
in biology.
● Some of the common applications include:
● Protein Contamination Detection on Surfaces
● Protein Detection and Identification
● Detection of Dangerous Biological Compounds
● Qualitative & quantitative estimation of minerals,
vitamins, enzyme kinetics, drug response, toxic
chemicals, nucleic acids.

17. Future of UV-Vis Spectrophotometers
● Future improvements in UV-Vis
spectrophotometers will focus on ease-
of-use, portability, and application-
specific instruments. UV-Vis analysis of
solid samples and materials continues to
grow in areas such as solar cell research,
semiconductor products, and coating
materials.
● Advances in light sources will provide
new developments in conventional
spectrophotometers and handheld UV-
Vis instruments.
● Further development in remote sensors
will enable more types of samples to be
measured outside the laboratory.

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