Absorbance spectroscopy uses the wavelength-dependent absorption characteristics of materials to identify and quantify specific substances ranging from the visible light to ultraviolet regions. It has various applications in natural product research including determining protein and nucleic acid concentrations by measuring absorbance according to the Lambert-Beer law. It can also monitor enzyme kinetics by measuring changes in absorbance over time. Absorbance spectroscopy is useful for real-time monitoring of bioprocesses and fermentation by measuring optical density and absorbance of biomass and cellular components. It has been used to evaluate phenolic compounds during winemaking.
Uv-Vis spectroscopy: electronic spectroscopy, absorption and emission, Terms describing UV absorptions, absorbing species containing s,n and pi, absorbing species,sigma and pi orbitals, electronic transitions, Absorption: physical Basis and lineshape,UV-Spectra.
Crown ethers
NOMENCLATURE
GENERAL SYNTHESIS OF CROWN ETHER
AZA CROWN
CRYPTAND
APPLICATIONS
1. SYNTHETIC APPLICTION
Esterification
Saponification
Anhydride formation
Potassium permanganate oxidation
Aromatic substitution reactions
Elimination reactions
Displacement reaction
Generation of carbenes
Superoxide anion
Alkylations – 1. o-alkylations
2. c-alkylations
3. n-alkylations
2. ANALYTICAL APPLICATION
Determination of gold in geological samples
Super critical fluid extraction of trace metal from solid and liquid materials
Application of ionic liquids in analytical chemistry
Oxidation and determination of aldehydes
Crown ethers are used in the laboratory as phase transfer catalyst
OTHER APPLICATION
It is used in photocynation
Resolution of racemic mixture
Benzoin condensation
Hetrocyclisation
Synthesis of furanones
Acetylation of secondary amines in presence of primary amine
Uv-Vis spectroscopy: electronic spectroscopy, absorption and emission, Terms describing UV absorptions, absorbing species containing s,n and pi, absorbing species,sigma and pi orbitals, electronic transitions, Absorption: physical Basis and lineshape,UV-Spectra.
Crown ethers
NOMENCLATURE
GENERAL SYNTHESIS OF CROWN ETHER
AZA CROWN
CRYPTAND
APPLICATIONS
1. SYNTHETIC APPLICTION
Esterification
Saponification
Anhydride formation
Potassium permanganate oxidation
Aromatic substitution reactions
Elimination reactions
Displacement reaction
Generation of carbenes
Superoxide anion
Alkylations – 1. o-alkylations
2. c-alkylations
3. n-alkylations
2. ANALYTICAL APPLICATION
Determination of gold in geological samples
Super critical fluid extraction of trace metal from solid and liquid materials
Application of ionic liquids in analytical chemistry
Oxidation and determination of aldehydes
Crown ethers are used in the laboratory as phase transfer catalyst
OTHER APPLICATION
It is used in photocynation
Resolution of racemic mixture
Benzoin condensation
Hetrocyclisation
Synthesis of furanones
Acetylation of secondary amines in presence of primary amine
ELECTRICAL DOUBLE LAYER-TYPES-DYNAMICS OF ELECTRON TRANSFER-MARCUS THEORY-TUNNELING - BUTLER VOLMER EQUATIONS-TAFEL EQUATIONS-POLARIZATION AND OVERVOLTAGE-CORROSION AND PASSIVITY-POURBAIX AND EVAN DIAGRAM-POWER STORAGE-FUEL CELLS
In molecular spectroscopy, a Jablonski diagram is a diagram that illustrates the electronic states of a molecule and the transitions between them. The states are arranged vertically by energy and grouped horizontally by spin multiplicity.
MASS SPECTROSCOPY ( Molecular ion, Base peak, Isotopic abundance, Metastable ...Sachin Kale
CONTENT:
Molecular Ion Peak
Significance of Molecular ion & Graphically Method
Base Peak
Isotopic Abundance
Metastable Ion
Significance of Metastable ion
Nitrogen Rule & graphs
Formulation of Rule
ELECTRICAL DOUBLE LAYER-TYPES-DYNAMICS OF ELECTRON TRANSFER-MARCUS THEORY-TUNNELING - BUTLER VOLMER EQUATIONS-TAFEL EQUATIONS-POLARIZATION AND OVERVOLTAGE-CORROSION AND PASSIVITY-POURBAIX AND EVAN DIAGRAM-POWER STORAGE-FUEL CELLS
In molecular spectroscopy, a Jablonski diagram is a diagram that illustrates the electronic states of a molecule and the transitions between them. The states are arranged vertically by energy and grouped horizontally by spin multiplicity.
MASS SPECTROSCOPY ( Molecular ion, Base peak, Isotopic abundance, Metastable ...Sachin Kale
CONTENT:
Molecular Ion Peak
Significance of Molecular ion & Graphically Method
Base Peak
Isotopic Abundance
Metastable Ion
Significance of Metastable ion
Nitrogen Rule & graphs
Formulation of Rule
The use of agrochemicals has increased considerably in recent years, and consequently, there has been increased exposure of ecosystems and human populations to these highly toxic compounds. The study and development of methodologies to detect these substances with greater sensitivity has become extremely relevant. This article describes, for the first time, the use of atomic force spectroscopy (AFS) in the detection of enzyme-inhibiting herbicides. A nanobiosensor based on an atomic force microscopy (AFM) tip functionalised with the acetolactate synthase (ALS) enzyme was developed and characterised. The herbicide metsulfuron-methyl, an ALS inhibitor, was successfully detected through the acquisition of force curves using this biosensor. The adhesion force values were considerably higher when the biosensor was used. An increase of ~250% was achieved relative to the adhesion force using an unfunctionalised AFM tip. This considerable increase was the result of a specific interaction between the enzyme and the herbicide, which was primarily responsible for the efficiency of the nanobiosensor. These results indicate that this methodology is promising for the detection of herbicides, pesticides, and other environmental contaminants.
Enzyme based Biosensor for pesticide DetectionSubhasis Sarkar
The biosensors could be used for pesticides rapid detection with a good stability and repeatability. As a new analytical method, biosensor could be widely used in the determination of food contamination. Biosensor techniques based on the principle of specific biological-recognition have shown satisfactory results for environmental control, food quality monitoring and toxicity detection in recent years. All these detection methods based on biosensors were shorter time response and lower cost comparing with the traditional method, but these methods were not enough convenient to use, moreover, complex detection procedures make them unsuitable for commercial and industrial applications.
A sensor that integrates a biological element with a physiochemical transducer to produce an electronic signal proportional to a single analyte which is then conveyed to a detector.
Food safety ( Basic steps in detection of food borne pathogens )SurbhiRai8
It consists of basic structure of steps for analysis of food borne pathogens in various ways and about these ways . what do we mean by food borne pathogens and why there is a need for their detection . then it has a little brief about each and every method . then we have covered 4 basic pathogens found in food and their detection methods . we are very thankful for all the sources from which we got this data . some of them are research papers and google books but it helped us to learn more .
Atomic force spectroscopy ~AFS! was used to measure interaction forces between the tip and nanostructured layers of poly~o-ethoxyaniline! ~POEA! in pure water and CuSO4 solutions. When the tip approach and retraction were carried out at low speeds, POEA chains could be physisorbed onto the Si3N4 tip
via nonspecific interactions.We conjecture that while detaching, POEA chains were stretched and the estimated
chain lengths were consistent with the expected values from the measured POEA molecular weight. The effects
from POEA doping could be investigated directly by performing AFS measurements in a liquid cell, with the
POEA film exposed to liquids of distinct pH values. For pH 6.0, the force curves normally displayed an
attractive region for POEA, but at lower pH values—where POEA is protonated—the repulsive double-layer
forces dominated. Measurements in the liquid cell could be further exploited to investigate how the film
morphology and the force curve are affected when impurities are deliberately introduced in the liquid. The
shape of the force curves and the film morphology depended on the concentration of heavy metal in the liquid
cell. AFS may therefore be used to study the interaction between film and analyte, with important implications
for the understanding of mechanisms governing the sensing ability of taste sensors.
Similar to Application of UV-vis in natural product research (20)
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2. Absorbance Spectroscopy
Absorbance spectroscopy is a
molecular spectroscopy method that uses the
wavelength-
dependent absorption characteristics of
materials to identify and quantify specific
substances. It ranges from Visible light region
to Ultraviolet region; 200-400nm. It is also
known as UV/Vis spectroscopy.
3. Natural Products
A natural product is a chemical compound or
substance produced by a living organism or
which can be prepared by chemical synthesis
by life. For instance, biotic materials (wood,
silk), bio-based materials (bioplastics,
cornstarch), bodily fluids (milk, plant
exudates), and other natural materials (soil,
coal).
4. Applications in Natural Product
Analysis
Absorption spectroscopy is an excellent
technique for following ligand-binding
reactions, enzyme catalysis and
conformational transitions in proteins and
nucleic acids.
Here follows the broad significance of
absorption spectroscopy in the areas of natural
product research.
5. Protein Concentrations
The concentrations of proteins or nucleic acids
in solution can be easily and accurately
determined by absorbance measurements
following the LAMBERT-BEER law;
A= -log10(I/I0) OR A=ecl
6. Protein Concentrations
Absorption coefficients of proteins
Proteins usually show absorption maxima
between 275 and 280nm, caused by the
absorbance of the two aromatic amino acids
tryptophan and tyrosine and, to a small extent,
by the absorbance of cystine (disulfide bonds)
7. Protein Concentrations
The absorbances of Trp and Tyr depend on
the microenvironment of their chromophores,
and they are slightly red-shifted when
transferred from a polar to a nonpolar
environment, such as in the interior of a
globular protein.
8. Protein Concentrations
The absorption coefficienteof a protein can be
calculated in a simple fashion. First the
numbers of its Trp, Tyr and Cys disulfide
bonds (nTrp, nTyr and nSS, respectively) are
counted, and then e calculated by use of
lambert beer law and the values obtained are:
5500, 1490, and 125 L/molcm respectively.
These represent average values for the
chromophores in folded proteins.
9. Absorbance of Proteins
The peptide groups of the protein main chain
absorb light in the ‘far-UV’ range (180–30nm).
10. Absorbance of Proteins
The aromatic side chains of Tyr, Trp and
Phe also absorb light in this region and, in
addition, they absorb in the 240–300nm
region. This region is called the ‘near-UV’ or
the ‘aromatic’ region.
Disulfide bonds that form between two
cysteine residues also show an absorbance
band near 260nm. Many cofactors absorb light
in the UVvis
11. (NADH) and reduced flavin–adenine
dinucleotide (FADH2) show spectra in the
near-UV.
Haem groups and copper-containing
cofactors absorb in the visible region.
Therefore haemoglobin is red and
plastocyanin is blue.
12. Concentrations of nucleic acids
The concentrations of nucleic acids in solution
are routinely determined from their strong
absorbance at 260nm.
Proteins absorb much more weakly than
nucleic acids. Contaminating proteins
therefore hardly affect the concentrations of
nucleic acids.
13. Absorbance of Nucleic Acids
Nucleic acids show a strong absorbance in the
region of 240-275nm. It originates from the pi
to pi* transitions of the pyrimidine and purine
ring systems of the nucleobases.
In native DNA the bases are stacked in the
hydrophobic core of the double helix and
accordingly their absorbance is considerably
decreased relative to the absorbance of single
stranded DNA.
14. The decrease in absorbance upon base
stacking in the interior of DNA or RNA double
helices is called hypochromism. It provides a
very sensitive and convenient probe for
monitoring strand dissociation and unfolding
(‘melting’) of DNA double helices.
15. Applications in Enzyme Kinetics
An enzyme catalyses the conversion of one or
several substrates to one or several products.
The rate of the catalysed reaction or the
activity of the enzyme can be determined by
measuring either the decrease in substrate
concentration or the increase in product
concentration as a function of the reaction
time.
16. When the substrate (S) and the product (P)
differ in absorbance, the progress of an
enzymatic reaction can be followed directly by
monitoring the change in absorbance as a
function of time.
The absorbance changes are linearly related
with the changes in concentration (via the
Lambert–Beer relation)
17. Therefore,the reaction rates can be
calculated from the absorbance data if
absorption coefficients of the reacting species
are known.
NADH-linked enzyme reactions, such as
those catalysed by the lactate,
dehydrogenases provide excellent examples
for absorbance-based enzyme assays. NADH
shows an absorbance maximum near 340nm.
18. UV-Vis Spectrophotometric Methods for
Methotrexate Assay
Methotrexate (MTX) is a well-known
anticancer drug used in the chemotherapy of
several malignant diseases, such as acute
lymphocytic leukemia, osteosarcoma, breast
and bladder cancers.
19. UV-Vis Spectrophotometric Methods for
Methotrexate Assay
UV/VIS spectroscopy for prolonging the drug
delivery has been used. However, the
evaluation of the drug content is still a
challenge for researchers.
An analytical curve is generated by plotting the
area under the curve or the absorbance for
UV-Vis spectrophotometric method.
20. Any change in the absorbance intensity and
any bathochromic or hypsochromic shift are
investigated by analyzing the impurities curve
/peaks in the absorbance graph.
If there are any impurities found by the shifts of
the spectrum, these are easily recognized and
reported.
21.
22. The Use in Bioprocess and
Fermentation Monitoring
A bioprocess is a specific process that uses
complete living cells or their components
(bacteria, enzymes) to obtain desired
products.
Fermentation is a metabolic process that
produces chemical changes and the extraction
of energy from carbohydrates in the absence
of oxygen by Bacteria.
23. There is an increasing need for real-time
analytical tools to monitor bioprocess and
fermentation in biological and food
applications.
Spectroscopic sensors, when combined with
PAT, enable simultaneous, real-time
bioprocess monitoring of parameters like
biological, chemical, and physical variables
during the process.
24. The development and implementation of these
spectroscopy methods in the field of food
analysis are based on the interactions
between matter and light that resulted in
absorption, emission, and scattering events
characteristic of the sample.
25. Therefore, on the basis of the absorption
measurement, the presence and concentration
of analytes in the food matrix as a
consequence of its chemical and physical
properties can be determined and quantified.
26. In bioprocess and fermentation monitoring,
optical density (OD) provides the most relevant
information to make the measurements. This
effect can be used to determine the
concentration of biomass in turbid samples.
27. For example, in the wavelength range
between 350 and 400 nm, several researchers
have reported the usefulness of this range to
differentiate between viable and dead
microbial cells due to an increase in
absorption associated with microbial cell
contents and nucleic acids originating from the
damaged microbial cells.
28. From the collected data, the information can
be visualized as a pattern that contains
information about a complex bioprocess—for
example, fermentation.
29. Phenolic Compounds
Evaluation
Phenolic compounds play an important role in the
colour, flavour and “mouth feel” attributes of
wines. Consequently, the measurement of
phenolic compounds during fermentation is
important to better understand and control the
winemaking process . UV-Vis spectroscopy was
used to monitor the phenolic composition during
winemaking, where models used to predict
phenolic compounds were developed. The
accuracy and robustness of the calibration models
were evaluated using the slope and intercept,
interclass correlation coefficients and standard
error of measurement.
30. Conclusion
The advent of new types of UV-Vis
instrumentation and sample presentation options
has aided in the development of new possibilities
to monitor many processes in biological samples.
The literature reports several considerable
successes to this end, particularly for the routine
screening of typical constituents, such as
anthocyanins, phenolics, sugars, and
antioxidants, and complex food matrices. The
challenges of isolating matrix interferences remain
but the incorporation of data mining and data
analysis techniques extends the possibilities of
using UV-Vis spectroscopy in natural products
analysis and quantification.