Research

Techniques in
Plants
COURSE INSTRUCTOR:
MS. QURAT UL AIN
PRESENTATION BY GROUP (1):
 ABDULLAH SALEEM (12)
 SAMREEN AKHTAR (20 )
 MUHAMMAD MURSALEEN (06)
 AYESHA KOSER (03)
 KAINAT RASOOL (13)
 IQRA AMIN (14)
Class:
 MS BOTANY 2ND SEMESTER
TOPIC:

SPECTROPHOTOMETRY
CONTENTS:
 INTRODUCTION
 SPECTROPHOTOMETER
 COMPONENTS OF SPECTROPHOTOMETER
 TYPES OF SPECTROPHOTOMETER
 APPLICATIONS OF SPECTROPHOTOMETER
 ADVANTAGES AND
LIMITATIONS OF
SPECTROPHOTOMETER
SPECTROPHOTOMETRY:

 Spectrophotometry is a method to measure how

much a chemical substance absorbs light (at
different wavelengths).

 It can be measure by measuring the intensity of

light as the beam of light passes through a
sample solution.
SPECTROPHOTOMETRY:

 This
depends on the concentration of that
chemical substance.

 Thebasic principle is that each compound

absorbs light over a certain range of wavelength.
SPECTROPHOTOMETER:

 Spectrophotometer is an instrument that is use

for spectrophotometry.

 Itmeasures the intensity of light after it passes

through a sample solution.
SPECTROPHOTOMETER:
COMPONENTS OF
SPECTROPHOTOMETER:
 1. Light Source: A spectrophotometer begins
with a light source that emits a broad spectrum
of light.
 2. Collimator:
 Itbends light so that photons travel parallel to
each other.
COMPONENTS OF
SPECTROPHOTOMETER:
 [Link]: Before the light hits the
sample, it passes through a monochromator,
which separates the light into its component
wavelengths.
 4. Wavelength Selector: The user selects the
specific wavelength of light that is known to be
absorbed by the sample of interest
COMPONENTS OF
SPECTROPHOTOMETER:
 5. Sample Holder or Cuvette: It's usually a
transparent container called a cuvette, made of
glass or quartz, designed to hold the sample liquid.
 6. Detector: The detector measures the intensity of
light after it passes through the sample.
 7. Digital Display unit: a digital display to show
the final calculations and results.
COMPONENTS OF
SPECTROPHOTOMETER:
TYPES OF SPECTROPHOTOMETER:

 Visible Spectrophotometer:
 An instrument used to measure absorbance and
conduct quantitative analysis at the visible light
(400 ~ 760nm), known as the visible
spectrophotometer. Bacterial cell density can be
determined at 600 nm.
Visible Spectrophotometer
TYPES OF SPECTROPHOTOMETER:
 UV-VIS Spectrophotometer
TYPES OF SPECTROPHOTOMETER:
 UV-VIS Spectrophotometer
 UV-VIS spectrophotometer is used to measure the
material of absorbance and quantitative analysis at the
visible or ultraviolet light (200 ~ 760nm). Nucleic acid
and protein concentrations can be measured and bacterial
cell density can also be determined. UV
spectrophotometer can be divided into single beam, split
beam, double beam for different applications.
 UV spectrophotometer can be divided into single beam,
split beam, double beam for different applications.
TYPES OF SPECTROPHOTOMETER:

 Infrared Spectrophotometer:
TYPES OF SPECTROPHOTOMETER:
 Infrared Spectrophotometer:
 The general infrared spectrum refers to the infrared spectrum
greater than 760nm, which is the most commonly used spectral
region of organic compounds, can analyze a variety of conditions
(gas, liquid, solid) of the sample.
 Infrared spectroscopy is characterized by fast, low sample
volume (a few micrograms to a few milligrams), strong
characterization (various substances have their own specific
infrared spectrum), tests capable of analyzing various states (gas,
liquid, solid) without damaging the sample.
TYPES OF SPECTROPHOTOMETER:
 Fluorescence Spectrophotometer:
TYPES OF SPECTROPHOTOMETER:
 Fluorescence Spectrophotometer:
 Fluorescence spectrophotometers are instruments used to scan the
fluorescence spectrum emitted by liquid fluorescent labels, which
is widely used in scientific research, chemical industry, medicine,
biochemistry, environmental protection, clinical testing, food
testing, teaching experiments and other fields.
 Through the determination of these parameters, the equipment is
not only for general quantitative analysis, but also can infer the
molecular conformational changes in various environments, thus
clarifying the relationship between molecular structure and
function.
TYPES OF SPECTROPHOTOMETER:
 Atomic absorption Spectrophotometer:
 The method is mainly applied to detect trace components in the
sample analysis. It is a powerful tool for material analysis and
elemental analysis of trace metals (semimetals).
APPLICATIONS OF
SPECTROPHOTOMETER
1. Chemical Analysis:
 Spectrophotometry is widely used in chemistry for
quantitative analysis of substances. It's employed in
determining the concentration of a solute in a solution
by measuring the absorbance of light at specific
wavelengths.
 This is vital in fields like pharmaceuticals,
environmental monitoring, and food and beverage
industries.
APPLICATIONS OF
SPECTROPHOTOMETER
2. Biochemical Assays:
 In biochemistry, spectrophotometry is crucial for
studying biomolecules like proteins, nucleic acids, and
enzymes.
 It'sused to quantify biomolecules, measure enzyme
activity, and determine DNA purity and concentration.
For instance, the Bradford assay utilizes
spectrophotometry to quantify protein concentrations.
APPLICATIONS OF
SPECTROPHOTOMETER
3. Medical Diagnostic:
 Spectrophotometers play a significant role in medical
diagnostics, including clinical chemistry and microbiology.
 They'reused to analyze blood samples for various
components like hemoglobin, glucose, and cholesterol.
Additionally, spectrophotometric methods are employed in
DNA analysis, such as polymerase chain reaction (PCR)
and DNA sequencing.
APPLICATIONS OF
SPECTROPHOTOMETER
4. Environmental monitoring:
 Spectrophotometry is instrumental in environmental
science for monitoring pollutants and assessing water
quality.
 It'sused to measure the concentration of pollutants like
heavy metals, pesticides, and organic compounds in air,
water, and soil samples. UV-visible spectrophotometry
is often used for such analyses.
APPLICATIONS OF
SPECTROPHOTOMETER
5. Food and beverage industry:
 Spectrophotometers are employed in food and beverage
analysis for quality control and safety assessment.
 They're used to measure parameters such as color,
acidity, sugar content, and vitamin concentration. This
ensures compliance with regulatory standards and
maintains product consistency.
APPLICATIONS OF
SPECTROPHOTOMETER
6. Pharmaceutical Analysis:
 Spectrophotometry is extensively used in
pharmaceutical analysis for drug development, quality
control, and formulation studies.
 It's employed to quantify drug concentrations, assess
purity, and study drug stability under different
conditions. UV-visible and infrared spectrophotometers
are commonly utilized in this field.
APPLICATIONS OF
SPECTROPHOTOMETER
7. Forensic Science:
 In forensic analysis, spectrophotometry is used for
analyzing substances like drugs, fibers, and bodily
fluids found at crime scenes.
 Ithelps identify and quantify trace evidence, aiding in
criminal investigations and court proceedings.
APPLICATIONS OF
SPECTROPHOTOMETER
8. Material Science:
 Spectrophotometers are utilized in material science for
studying the optical properties of materials, such as
reflectance, transmittance, and absorbance.
 This information is crucial for developing new materials
with specific optical characteristics for applications like
optical coatings, sensors, and photovoltaic devices.
ADVANTAGES OF
SPECTROPHOTOMETERS:
 1. Accuracy: They provide precise measurements of the
absorbance or transmittance of a sample across a range
of wavelengths.
 2. Versatility:
Spectrophotometers can analyze various
types of samples, including liquids, gases, and solids
 [Link]: They can detect minute quantities of
substances, making them valuable in fields like
environmental monitoring, pharmaceuticals, and
biochemistry.
ADVANTAGES OF
SPECTROPHOTOMETERS:
 4. Speed:
 Modern spectrophotometers can provide results rapidly,
allowing for high-throughput analysis in research and
industrial settings.
 5. Quantitative Analysis:
 They enable quantitative analysis of substances,
allowing researchers to determine concentrations
accurately.
ADVANTAGES OF
SPECTROPHOTOMETERS:
 6. Wide Range of Applications:
 Spectrophotometry finds applications in diverse fields
such as chemistry, biology, physics, environmental
science, and materials science.
 7. Non-destructive:
 Spectrophotometric analysis typically doesn't alter the
sample, preserving its integrity for further testing or
analysis.
LIMITATIONS OF
SPECTROPHOTOMETERS:
1. Limited Wavelength Range
2. Limitations in Sample Concentration
3. Interference from other compounds
4. Limited to compounds that absorb light
 Any
Question?
THANK YOU