This document discusses photometry, colorimetry, and spectrophotometry, including their principles, instrumentation, and applications. Photometry measures light absorption to determine concentrations of elements. Colorimetry uses the human eye to determine concentrations of colored compounds, following Beer's and Lambert's laws relating absorbance to concentration and path length. Spectrophotometry identifies compounds by their absorption spectra and is used for enzyme assays, molecular weight determination, and studying protein interactions. These techniques are important in biochemical research for concentration determination, reaction monitoring, and compound identification.
It is the most common analytical technique used in biochemical estimation in clinical laboratory.
It involves the quantitative estimation of color.
A substance to be estimated colorimetrically, must be colored or it should be capable of forming chromogens (colored complexes) through the addition of reagents.
It is the most common analytical technique used in biochemical estimation in clinical laboratory.
It involves the quantitative estimation of color.
A substance to be estimated colorimetrically, must be colored or it should be capable of forming chromogens (colored complexes) through the addition of reagents.
Centrifugation principle and types by Dr. Anurag YadavDr Anurag Yadav
concept of cnetrifugation,
basic Principle
centrifugal force
types of centrifugation based on use and rotor type
application of the each type of centrifuge
Ultracentrifuge in detail
application in general
Colorimeter and spectrophotometer, Mass Spectrometerprachann
It contains a brief knowledge on Introduction, Principle, Laws, Flow representation, Instrumentation, Applications
and Mass spectrometer
- Principle
- Instrumentation
Introduction
History
Elecrophoresis
Principle
Types of electrophoresis
Application
Conclusion
Reference
When a potential difference is applied between the two electrodes in a colloidal solution, It has been observed that the colloidal particles are carried to either the positive or negative electrode.
In other words , they behave as if they are electrically charged w.r.t. the dispersion medium. This phenomenon is known as electrophoresis.
Many important biological molecules, such as amino acids, peptides, proteins, nucleotides and nucleic acids, possess ionisable groups and, therefore, at any given pH, exist in solution as electrically charged species either as cations or anions.
Under the influence of an electric field these charged particles will migrate either to the cathode or to the anode, depending on the nature of their net charge.
Centrifugation principle and types by Dr. Anurag YadavDr Anurag Yadav
concept of cnetrifugation,
basic Principle
centrifugal force
types of centrifugation based on use and rotor type
application of the each type of centrifuge
Ultracentrifuge in detail
application in general
Colorimeter and spectrophotometer, Mass Spectrometerprachann
It contains a brief knowledge on Introduction, Principle, Laws, Flow representation, Instrumentation, Applications
and Mass spectrometer
- Principle
- Instrumentation
Introduction
History
Elecrophoresis
Principle
Types of electrophoresis
Application
Conclusion
Reference
When a potential difference is applied between the two electrodes in a colloidal solution, It has been observed that the colloidal particles are carried to either the positive or negative electrode.
In other words , they behave as if they are electrically charged w.r.t. the dispersion medium. This phenomenon is known as electrophoresis.
Many important biological molecules, such as amino acids, peptides, proteins, nucleotides and nucleic acids, possess ionisable groups and, therefore, at any given pH, exist in solution as electrically charged species either as cations or anions.
Under the influence of an electric field these charged particles will migrate either to the cathode or to the anode, depending on the nature of their net charge.
This content is suitable for medical technologists/technicians/lab assistants/scientists writing the SMLTSA board exam. The content is also suitable for biomedical technology students and people also interested in learning about test methodologies used in medical technology. This chapter describes test methodologies and their uses. Please note that these notes are a collection I used to study for my board exam and train others who got distinctions using these.
Disclaimer: Credit goes to those who wrote the notes and the examiners of each exam question. Please use only as a reference guide and use your prescribed textbook for the latest and most accurate notes and ranges. The material here is not referenced as it is a collection of pieces of study notes from multiple people, and thus will not be held viable for any misinterpretations. Please use at your own discretion.
INSTRUMENTAL METHODS OF ANALYSIS, B.PHARM 7TH SEM. AND FOR BSC,MSC CHEMISTRY. This is Geeta prasad kashyap (Asst. Professor), SVITS, Bilaspur (C.G) 495001
Normal Labour/ Stages of Labour/ Mechanism of LabourWasim Ak
Normal labor is also termed spontaneous labor, defined as the natural physiological process through which the fetus, placenta, and membranes are expelled from the uterus through the birth canal at term (37 to 42 weeks
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
1. PHOTOMETRY , COLORIMETRY AND
SPECTROPHOTOMETRY , THEIR APPLICATION.
Presented by :-
Gadhavi Shital N.
Msc.Sem -2 (Botany)
Paper :- 406
Department of life sciences
H.N.G.U Patan
2. CONTENT
Photomtery principle
Principle of operation
Uses of photometry
Colorimetry principle
Principle of operation
spectrophotometry application
Principle of operation
Limitations
Uses of colorimetry and spectrophotometry
3. PHOTOMETRY
Principle
• The word is composed of the Greek word photo-
“light” and metry – “measure”
• The basis of photometric working is that , the species
of alkali metals and alkaline earth metals are
dissociated due to the thermal energy provided by the
flame source.
• Due to this thermal excitation, some of the atoms are
exited to a higher energy level where they are not
stable.
• The absorbance of light due to the electrons excitation
can be measured by using the direct absorption
techniques .
4. • The subsequent loss of energy will result in the
movement of exited atoms to the low energy ground
state with emission of some radiations, which can be
visualized in the visible region of the spectrum.
• The absorbance due to the electron excitation can be
measured by using the direct absorption technique
while the emititing radiation intensity is measured
using the emission techniques .
• The wavelength of emitted light is specific for specific
elements .
7. Uses of photometry
• Photometry is used in various industries like
chemicals, soils , agriculture, pharmaceuticals, glass,
and ceramics, in plant materials and water , and in
biological and microbiological laboratories .
• It is used in determination of potassium, sodium,
magnesium and calcium in biological fluids like serum,
plasma, urine etc , is routinely carried out by
photometer .
• Analysis of industrial water natural water for
determining elements responsible for hard water is
standard procedure in many laboratories.
8. COLORIMETRY
• Colorimetry is the use of the human eye to
determine the concentration of colored species.
• Colorimetric methods are applicable to dilute
solutions.
• For a colorimetric method to be quantitative , it must
from a compound with definite color
characterisitics.
• Color amount must be directly propotional to the
concentration .
• Colored compound must obey beer’s law and
lambert’s law.
9. PRINCIPLE
Beer’s Law: The amount of light absorbed by a
substance in a solution is proportional to a number
of absorbing molecules… i.e. concentration of the
absorbing solution
Log e
𝐼˳
𝐼
=kc
I=I˳𝑒−𝑘𝑐
k = constant for particular solution
c = concentration
10. • Lambert’s law :The amount of light absorbed by a
substance in a solution is proportional to the
thickness of the absorbing molecules …i.e . path
length
Log e
𝐼˳
𝐼
=kd
I=I˳ 𝑒−𝑘𝑑
d= length of absorbing layer
k = constant for particular solution
11. • Visual observations :- because colorimetry is
based on inspection of materials with the
human eye, it is necessary to review aspects
of visible light.
• Visible light is the narrow range of
electromagnetic waves with the wavelength
of 400-700 nm.
• Visible light is only small portion of the
electromagnetic spectrum.
14. SPETROPHOTOMETRY
Applications
Qualitative analysis : used to identify classes of
compound in both pure state and in biological
preparations. This is based on the fact that
absorption spectra are specific for a class of
compound.
Denatration of double standard DNA : DNA absorbed
at 260 nm. This absorbance increase with rise in
temperature , as double standard DNA become
single stranded.
15. Continues….
• Enzyme assay and kinetic studies : the
quantitative assay of enzyme activity is carried
out when substrate or product is colored
absorbs light in UV range.
• Molecular weight determination : the
molecular weight of amines , , sugars and
many aldehyde and ketone compounds can be
determined .
16. Continues…..
• Control purity : impurity in compound can be
detected easily spectrometric studies
• 1, carbon disulfide impurity in carbon
tetrachloride can be detected easily by
measuring absorbance at 318 nm where only
carbon disulfide absorb.
• Protein folding: protein or protein– nucleic
acid interaction can be studies
20. Limitation
• Applicable for monochromatic light.
• Applicable lower concentration.
• Applicable only clear solution.
21. • Colorimetry and spectophotometry are of prime
importance in biochemical research .
If the absorbancy index at a specific wavelength is known,
the concentration of a compound can be readily
determind by measuring optical density at that
wavelength . As the value of absorbancy index is very large
in nucleotides, very small quantities of the absorbing
material can be accurately measured.
The course of a reaction can be determind by measuring
the rate of formation or disappearance of a light absorbing
substance .thus NADH absorbs strongly at 340 nm–
whereas the oxidised form NAD—has no absorption at this
wavelength . Therefore, reactions involving the production
or utilisation of NADP can be assayed by the technique .
22. Compound can be frequently identified by
determining the characteristic absorption
spectra in the ultraviolet and visible region of
the spectrum.