*Nuclear magnetic resonance spectroscopy, most

commonly known as NMR spectroscopy or magnetic

resonance spectroscopy (MRS), is
a spectroscopic technique to observe local magnetic
fields around atomic nuclei.  It is a spectroscopy
technique that is based on the absorption of
electromagnetic radiation in the radiofrequency
region 4 to 900 MHz by nuclei of the atoms. Over the
past fifty years, NMR has become the preeminent
technique for determining the structure of organic
compounds. Of all the spectroscopic methods, it is
the only one for which a complete analysis and
interpretation of the entire spectrum is normally
expected. *Principle of Nuclear Magnetic Resonance
(NMR) Spectroscopy The principle behind NMR is
that many nuclei have spin and all nuclei are
electrically charged. If an external magnetic field is
applied, an energy transfer is possible between the
base energy to a higher energy level (generally a
single energy gap). The energy transfer takes place at
a wavelength that corresponds to radio frequencies
and when the spin returns to its base level, energy is
emitted at the same frequency. The signal that
matches this transfer is measured in many ways and
processed in order to yield an NMR spectrum for the
nucleus concerned. *Applications of Nuclear
Magnetic Resonance (NMR) Spectroscopy
Spectroscopy is the study of the interaction of
electromagnetic radiation with matter. NMR
spectroscopy is the use of the NMR phenomenon to
study the physical, chemical, and biological
properties of matter.It is an analytical chemistry
technique used in quality control.It is used in
research for determining the content and purity of a
sample as well as its molecular structure. For
example, NMR can quantitatively analyze mixtures
containing known compounds.NMR spectroscopy is
routinely used by chemists to study chemical
structure using simple one-dimensional techniques.
Two-dimensional techniques are used to determine
the structure of more complicated molecules.These
techniques are replacing x-ray crystallography for
the determination of protein structure.Time domain
NMR spectroscopy techniques are used to probe
molecular dynamics in solution.Solid state NMR
spectroscopy is used to determine the molecular
structure of solids.Other scientists have developed
NMR methods-of measuring diffusion coefficients.
*Chemical shift The precise resonant frequency of
the energy transition is dependent on the effective
magnetic field at the nucleus. This field is affected
by electron shielding which is in turn dependent on
the chemical environment. As a result, information
about the nucleus' chemical environment can be
derived from its resonant frequency. In general, the
more electronegative the nucleus is, the higher the
resonant frequency. Other factors such as ring
currents (anisotropy) and bond strain affect the
frequency shift. It is customary to adopt
tetramethylsilane (TMS) as the proton reference
frequency. This is because the precise resonant
frequency shift of each nucleus depends on the
magnetic field used. The frequency is not easy to
remember (for example, the frequency of benzene
might be 400.132869 MHz) so it was decided to
define chemical shift as follows to yield a more
convenient number such as 7.17 ppm. δ = (ν-ν0)/ν0
The chemical shift, using this equation, is not
dependent on the magnetic field and it is convenient
to express it in ppm where (for proton) TMS is set to
ν0 thereby giving it a chemical shift of zero. For
other nuclei, ν0 is defined as Ξ νTMS where Ξ (Greek
letter Xsi) is the frequency ratio of the nucleus (e. g.,
25.145020% for 13C). In the case of the 1H NMR
spectrum of ethyl benzene the methyl (CH3) group is
the most electron withdrawing (electronegative) and
therefore resonates at the lowest chemical shift. The
aromatic phenyl group is the most electron donating
(electropositive) so has the highest chemical shift.
The methylene (CH2) falls somewhere in the middle.
However, if the chemical shift of the aromatics were
due to electropositivity alone, then they would
resonate between four and five ppm. The increased
chemical shift is due to the delocalized ring current
of the phenyl group.* Spin-spin couplingThe effective
magnetic field is also affected by the orientation of
neighboring nuclei. This effect is known as spin-spin
coupling which can cause splitting of the signal for
each type of nucleus into two or more lines.The size
of the splitting (coupling constant or J) is
independent of the magnetic field and is therefore
measured as an absolute frequency (usually Hertz).
The number of splittings indicates the number of
chemically bonded nuclei in the vicinity of the
observed nucleus. Spin-spin coupling is the
interaction between the spin magnetic moments of
different electrons and/or nuclei. In NMR
spectroscopy it gives rise to multiplet patterns, and
cross-peaks in two-dimensional NMR spectra.
Between electron and nuclear spins this is termed
the nuclear hyperfine interaction. Between electron
spins it gives rise to relaxation effects and splitting
of the EPR spectrum. Spin-spin coupling between
spinning nuclei. The interaction between the spin
magnetic moments of the different sets of H atoms in
the molecule under study, is known as spin-spin
coupling. It is imperative that a minimum of 2 sets
of protons are present in adjacent positions. The
magnetic spins of these resonating nuclei interact
with each other and affect each other’s precession
frequencies. The effective magnetic field (Beff)
experienced by neighboring protons as a result of
magnetic spins thereby affect the chemical shift
values. [5] In addition to the chemical shifts, the
nature of the peaks in the NMR spectrum is also
affected.* Grignard reagents are extremely useful
organometallic compounds in the field of organic
chemistry. They exhibit strong nucleophilic
qualities and also have the ability to form new
carbon-carbon bonds. Therefore, they display
qualities that are also exhibited by organolithium
reagents and the two reagents are considered
similar.When the alkyl group attached to a Grignard
reagent is replaced by an amido group, the resulting
compound is called a Hauser base. These
compounds are even more nucleophilic that their
Grignard counterparts.Grignard reagents are known
for their ability to quickly target carbonyls at their
carbon level. Grignard reagents do not, however,
function in the presence of protic solvents. Instead of
reacting with the desired molecule, the Grignard is
so unstable that it can readily accept a proton from
a protic solvent. The Grignard then is inert and there
is no reaction to the desired molecule. A Grignard
reagent is an organomagnesium compound which
can be described by the chemical formula ‘R-Mg-X’
where R refers to an alkyl or aryl group and X refers
to a halogen. Preparation of Grignard Reagents The
process of preparing Grignard reagents is described
in the points provided below. It can be noted that
many of these reagents can also be purchased
commercially.These reagents are prepared via the
treatment of magnesium with organic halides such
as alkyl or aryl halides. This is done with the help of
solvents comprising ethers (which are described by
the formula R-O-R’) because the ligands provided by
these solvents help in the stabilization of the
organomagnesium compound. Water and air are
very harmful to this synthesis and can quickly
destroy the Grignard reagent which is being formed
via protonolysis or via oxidation of the reagent.
Therefore, the process must be carried out in air-free
conditions. Alternatively, the magnesium can be
activated to make it consume water when wet
solvents are used with the help of ultrasound. After
the slow induction period of the reaction, the process
can be quite exothermic. This is a very important
factor to consider while industrially producing the
Grignard reagent. The organic halides used in these
reactions include aryl or alkyl chlorides, bromides,
and iodides. Aryl fluorides and alkyl fluorides are not
very reactive and are hence not commonly used.
However, with the help of Rieke metals, the
magnesium can be activated to make the fluoride
more reactive. Reactions of Grignard
ReagentsDuring a reaction involving Grignard
reagents, it is necessary to ensure that no water is
present which would otherwise cause the reagent to
decompose rapidly. Therefore, the majority of
Grignard reactions occur in solvents such as
anhydrous diethyl ether or tetrahydrofuran because
the oxygen in these solvents stabilizes the
magnesium reagent.Grignard reagents are very
important reagents in organic chemistry since they
can be reacted with a wide range of compounds to
form different products. Some of these reactions of
these reagents are listed below.1. Reactions with
Carbonyl Group These reagents form various
products when reacted with different carbonyl
compounds. The most common reaction of Grignard
reagents is the alkylation of ketones and aldehydes
with the help of R-Mg-X. 2. Reactions with Non-
Carbon ElectrophilesFor the formation of new
carbon-heteroatom bonds, Grignard reagents and
some organolithium compounds are very useful.
These reagents can also undergo a transmetallation
reaction with cadmium chloride, yielding dialkyl
cadmium. This reaction can be written as
follows.2R-Mg-X + CdCl2 → R2Cd + 2Mg(X)ClAlkyl
chains can be attached to many metals and
metalloids with the help of these reagents.3.
Reactions with Organic HalidesTypically, these
reagents are quite unreactive towards organic
halides which highly contrasts their behaviour
towards other halides. However, carbon-carbon
coupling reactions occur with Grignard reagents
acting as a reactant when a metal catalyst is
introduced.An example of such a coupling reaction
is the reaction between methyl p-chlorobenzoate and
nonyl magnesium bromide which yields the
compound p-nonyl benzoic acid in the presence of
the catalyst – Tris(acetylaceto) iron(III).4. Reaction
between Acetone and Methyl Magnesium
ChlorideThe reaction of methyl magnesium bromide
with acetone followed by hydrolysis gives tertiary
alcohol. Acetone reacts with methyl magnesium
bromide followed by hydrolysis to give secondary
alcohols..* Alkyl hydrogen atoms bonded to a carbon
atom in a  (alpha) position relative to a carbonyl
group display unusual acidity. While the pKa values
for alkyl C-H bonds is typically on the order of 40-
50, pKa values for these alpha hydrogens is more on
the order of 19-20. This can most easily be explained
by resonance stabilization of the product
carbanionIn the presence of a proton source, the
product can either revert back into the starting
ketone or aldehyde or can form a new product, the
enol. The equilibrium reaction between the ketone or
aldehyde and the enol form is commonly referred to
as "keto-enol tautomerism". The ketone or aldehyde
is generally strongly favored in this reaction.Because
carbonyl groups are sp2 hybridized the carbon and
oxygen both have unhybridized p orbitals which can
overlap to form the C=O π bond. The presence of
these overlapping p orbitals gives α hydrogens
(Hydrogens on carbons adjacent to carbonyls)
special properties. In particular, α hydrogens are
weakly acidic because the conjugate base, called an
enolate, is stabilized though conjugation with the π
orbitals of the carbonyl. The effect of the carbonyl is
seen when comparing the pKa for the α hydrogens of
aldehydes (~16-18), ketones (~19-21), and esters
(~23-25) to the pKa of an alkane (~50). Of the two
resonance structures of the enolate ion the one
which places the negative charge on the oxygen is
the most stable. This is because the negative change
will be better stabilized by the greater
electronegativity of the oxygen.* Mutarotation is a
change in the optical rotation of a solution due to a
change in the equilibrium between alpha (ɑ) and
beta (β) anomers, upon dissolution in the aqueous
solution.The process is also known as
anomerization. Mutarotation is the change in
specific rotation of a chiral compound due to
epimerization. The term is most commonly used in
carbohydrate chemistry.eg: The monosaccharide D-
glucose exists in two cyclic forms, α-D-glucose
([α]D25 = +112) and β-D-glucose ([α]D25 = +18.7),
which are epimers and are available as pure
compounds. When one of the cyclic forms of D-
glucose is added to water, it undergoes reversible
epimerization to the other via the open-chain form,
during which the specific rotation of the solution
changes gradually until it reaches the equilibrium
value +52.7º.* Epimerization is a process in which
the configuration of one chiral center changes. The
two are then called diastereomers of each other.*
Amino Acids are the organic compounds that
combine to form proteins, hence they are referred to
as the building components of proteins. These
biomolecules are involved in several biological and
chemical functions in a human body and are the
necessary ingredients for the growth and
development of human beings. There are about 300
amino acids that occur in nature.” Amino acids are
organic compounds containing the basic amino
groups (-NH2) and carboxyl groups (-COOH). The
ingredients present in proteins are of amino acids.
Both peptides and proteins are long chains of amino
acids. Altogether, there are twenty amino acids,
which are involved in the construction of proteins.
General properties of Amino acids They have a very
high melting and boiling point. Amino acids are
white crystalline solid substances.In taste, few
Amino acids are sweet, tasteless, and bitter.Most of
the amino acids are soluble in water and are
insoluble in organic solvents.Essential and Non-
essential Amino acidsOut of 20 amino acids, our
body can easily synthesize a few on its own and are
called non-essential amino acids. They include
alanine, asparagine, arginine, aspartic acid,
glutamic acid, cysteine, glutamine, proline, glycine,
serine, and tyrosine.Apart from these, there are
other nine amino acids, which are very much
essential as they cannot be synthesized by our body.
They are called as essential amino acids, and they
include Isoleucine, histidine, lysine, leucine,
phenylalanine, tryptophan, methionine, threonine,
and valineIn cells, DNA (Deoxyribonucleic acid) is
the nucleic acid that functions as the original
blueprint for the synthesis of proteins. DNA contains
the sugar deoxyribose, phosphates and a unique
sequence of the nitrogenous bases adenine (A),
guanine (G), cytosine (C) and thymine (T). The DNA
molecules contain instructions a living entity
requires to grow, develop and reproduce. These
instructions are present inside each cell and are
inherited from the parents to their offsprings.It is
made up of nucleotides which contain nitrogenous
group, a phosphate group, and a sugar group. The
order of the nitrogenous bases – thymine(T),
guanine(G), cytosine(C), and adenine(A), is crucial in
determining the genetic code.Genes are formed by
the order of the nitrogenous bases present in the
DNA which is crucial for protein synthesis. The RNA
is another nucleic acid that translates genetic
information into proteins from DNA.The nucleotides
are linked together for the formation of two long
strands which spiral to produce a structure known
as the double-helix which resembles that of a ladder
wherein the sugar and phosphate molecules form
the sides while the rungs are formed by the
bases.The bases located on one strand pair up with
the bases on the other strand, as in – guanine pairs
with cytosine and adenine pairs with thymine.The
DNA molecules are extremely long and hence
without the right packaging, they cannot fit into
cells. Thus, DNA is tightly coiled to produce
formations referred to as chromosomes. Every
chromosome has a single DNA molecule. In humans,
there are 23 pairs of chromosomes that are present
within the nucleus of the cells*Ribonucleic acid
(RNA) is a nucleic acid which is directly involved in
protein synthesis. Ribonucleic acid is an important
nucleotide with long chains of nucleic acid present
in all living cells. Its main role is to act as a
messenger conveying instructions from DNA for
controlling the proteins synthesis.RNA contains the
sugar ribose, phosphates, and the nitrogenous bases
adenine (A), guanine (G),  cytosine (C),  and uracil
(U). DNA and RNA share the nitrogenous bases A, G,
and C. Thymine is usually only present in DNA and
uracil is usually only present in RNA.* Dyes are the
chemical substances used to impart colour to
fabrics, foods, and other objects for their
beautification and distinction. They are capable of
getting fixed to the fabrics/objects permanently and
are resistant to the action of water, soap, light, acid
and alkalies. Acid Dyes- These are azo dyes and are
characterized by the salts of sulphonic or carboxylic
acids. These are usually applied to wool, silk and
nylon and have no affinity for cotton. Example-
Orange I, Orange II, Methyl Orange, Martius Yellow
and Naphthol Yellow.Basic Dyes- These dyes contain
Amino group in unsubstituted (-NH2) or Substituted
form (-NR2) as chromophore (colour bearing group)
or auxochrome (colour enhancing group). These form
water-soluble cations in an acid solution which then
react with anionic sites present on the fabrics and
thus get attached to them. These are used to dye
modified nylons, polyesters, paper, leather, wool,
cotton etc. Example- Aniline Yellow and Malachite
Green.Direct Dyes- These also belong to the class of
azo dyes and are used to dye the fabrics directly by
placing it in an aqueous solution of the dye. These
are suitable for those fabrics which can form H-
bonds with the dyes. These are used to dye wool,
silk, nylon, rayon and cotton. Example- Martius
Yellow and Congo Red. Disperse Dyes- These are
water-insoluble dyes which are dispersed in reagents
like Phenol, Cresol etc. before applying to synthetic
fibres. These are used to dye nylon, polyesters and
polyacrylonitrile. Example- Celliton Fast Pink B and
Celliton Fast Blue B.Fibre Reactive Dyes- These dyes
contain a reactive group which combines directly
with the hydroxyl or Amino group of the fibre.
Because of the irreversible chemical reaction, the
colour is fast and has a long life. These are used to
dye nylon, wool and silk.Insoluble Azo Dyes (Ingrain
Dyes)- These dyes are directly synthesized on the
surface of the fibre. The fabric to be coloured is
soaked in an alkaline solution of Phenol or Naphthol
and is then treated with a solution of diazotised
amine to produce azo dye on the surface of the
fabric. The colour imparted by such dyes is not very
fast. These are used to dye nylon, cotton, silk,
polyester etc. Example- Nitroaniline red.* A
heterocyclic compound has at least two different
elements as a member of its ring.The most common
hetero atoms found on a cyclic ring are Oxygen (O),
Nitrogen (N) and Sulphur (S).Example:Nucleic Acid,
present in the body responsible for storing and
expressing genetic information, is an example of a
Heterocyclic compound.Essential micronutrient,
Vitamins is also an example of a heterocyclic
compound.The majority of drugs, pesticides, dyes,
and plastics are examples of heterocyclic
compounds.Classification of Heterocyclic
CompoundBased on the electronic arrangement, we
can classify Heterocyclic compounds into two
types:Aliphatic Heterocyclic CompoundAromatic
Heterocyclic CompoundAliphatic Heterocyclic
CompoundAliphatic heterocyclic compounds are
those cyclic heterocycles that do not contain any
double bond.The properties of aliphatic heterocyclic
compounds are mainly affected due to ring
strain.Examples of aliphatic heterocyclic compounds
are Aziridine, Ethylene Oxide, Thiirane, Oxetane,
Azetidine, Thietane, Tetrahydrofuran (THF), Dioxane,
Pyrrolidine, Piperidine, etc.Aromatic Heterocyclic
CompoundAromatic heterocyclic compounds, as the
name suggests, are cyclic aromatic
compounds.Aliphatic Heterocyclic compounds obey
Huckels Rule, i.e.It should be cyclic.It should be
planar.It should not contain any sp3 hybridised
atoms.It must have (4n+2) 𝛑 electrons.Aromatic
Heterocyclic compounds are analogous to
Benzene.Examples: Furan, Pyrrole, Thiophene,
Indole, Benzofuran, Carbazole, Quinoline,
Isoquinoline, Imidazole, Oxazole, Pyrazole,
Pyridazine, Pyrimidine, Purine, etc.* Preparation
Methods By passing a mixture of acetylene and
ammonia through a red-hot tube 2. By heating
ammonium mucate with glycerol at 200 degrees. At
this temperature, ammonium mucate is dissociated
into mucic acid and ammonia. The acid then
undergoes dehydration, Physical Properties of
Pyrrole Pyrrole is colorless liquid, Boiling point
131°C, which rapidly turns brown on exposure to
air. Its odour is like that of chIoroform. Pyrrole is
sparingly soluble in water but dissolves in ethanol
and ether Chemical Properties of Pyrrole Basic
Character: Pyrrole reacts with dilute hydrochloric
acid to give a crystalline hydrochloride. Acidic
Character Pyrrole is not only a weak base but also a
very weak acid. This is shown by its reactions with
potassium hydroxide and Grignard reagents.
Electrophilic Substitution ReactionsPyrrole
undergoes electrophilic substitution reactions at C-2
because three resonance forms can be written for
the intermediate obtained from attack at C-2,
whereas only two such forms are possible for
substitution at C-3. Consequently the C-2
intermediate is more stable and the product with a
substituent at C-2 predominates. Substitution at C-
3 occurs only when both the 2-positions (that is, α
and α') are blocked decarboxylation and ring-closure
by reaction with ammonia.* Preparation Methods By
dry-distillation of mucic acid and heating the
product, furoic acid (furan-2-carboxylic acid), at
200-300°C.By oxidation of furfural with potassium
dichromate to give furoic acid and subsequent
decarboxylation at 200-300°C. By decarbonylation of
furfural in steam in the presence of silver oxide
catalyst (Commercial Method of Preparation). By
dehydration of succinic dialdehyde by heating with
P2O5 or ZnCl2.Physical properties of Furan Furan is
a colourless liquid, boiling point 32C, with a
chloroform like smell. It is only slightly soluble in
water, but dissolves in most organic solvents.
Chemical properties of Furan Furan is the most
reactive of all 5-membered heterocycles.. Basic
Character: Furan is a weak base like pyrrole. It
forms unstable salts with mineral acids. These salts
may either polymerize to produce a brown resin or
undergo hydrolysis to yield succindialdehyde. 2.
Electrophilic Substitution Like pyrrole, it undergoes
electrophilic substitution at C-2. Substitution at C-3
occurs only when both of the C-2 positions (α and ά)
are already blocked. Reduction: Furan is reduced by
hydrogen in the presence of nickel to produce
tetrahydrofuraN *Preparation Methods By passing a
mixture of acetylene and hydrogen cyanide through
a red- hot tube. By dehydrogenation of piperidine
with concentrated sulphuric acid at 300°C or with
nitrobenzene at 260°C. By heating
tetrahydrofurfuryl alcohol with ammonia in the
presence of aluminium oxide at 500°C. (Commercial
Method of Preparation). Physical properties of
Pyridine Pyridine is a colourless liquid, bp 115C°. It
has a very characteristic pungent and disgusting
odour. Pyridine is miscible with water and most
organic solvents. Almost all classes of organic
compounds are soluble in pyridine, even many of the
high melting solids which scarcely dissolve in
solvents such as ethanol and benzene. It is
consequently used as a solvent.It is very
hygroscopic. Chemical properties of Pyridine Basic
Character: Pyridine behaves as a base. It reacts with
acids to form fairly stable salts.. Electrophilic
Substitution: Pyridine, however, does undergo
electrophilic substitution reactions when extremely
vigorous reaction conditions are used. Substitution
occurs almost exclusively at C-3 (β-Position).
Pyridine does not undergo Friedel-Crafts acylation
and alkylation. This is because the Lewis acids (e.g.,
AlCl3) which are used as catalysts in these reactions
coordinate with the lone pair of electrons on
nitrogen. Nucleophilic Substitution: Pyridine
undergoes nucleophilic substitution reactions
mainly at C-2 (or at C-4 if C-2 is blocked) Reduction:
Pyridine undergoes reduction with lithium
aluminium hydride (LIAIH4), or hydrogen in the
preSence of nickel catalyst to form piperidine.
Oxidation: Like benzene, pyridine is quite stable
towards mild oxidizing agents. It does not react with
chromic acid or nitric acid. However, it may be
oxidized by peracetic acid to give pyridine-N-oxide.
*Preparation Methods By passing a mixture of
acetylene and hydrogen sulphide through a tube
containing aluminium oxide at 400°C. By heating
sodium succinate with phosphorous trisulplside By
the high-temperature (650 C) reaction of sulphur
with butane. (Commercial Method of Preparation).
Physical properties of Thiophene Thiophene is a
colourless liquid, boiling point 84 C, with an odour
very similar to that of benzene. It is insoluble in
water, but miscible with most organic solvents.
Chemical properties of Thiophene Thiophene is 300
times more reactive than benzene. Thiophene does
not show any basic properties. It is much more
stable to acids than either pyrrole or furan.
ThiopheneMdoes not undergo the Diels-Alder
reaction. Electrophilic Substitutions: Thiophene,
like furan and pyrrole, undergoes electrophilic
substitution reactions primarily at C-2. Substitution
at C-3 occurs only when both of the 2-positions (α
and ά) are already occupied. Electrophilic
Substitution Reduction: Thiophene may be
hydrogenated by means of sodium amalgam and
ethanol to tetrahydrothiophene. Desulphurisation:
Catalytic reduction of Thiophene with Raney Nickel
results in the removal of Sulphur to form n-
butane*Acid value is defined as the amount of
KOH needed in milligrams to neutralize the organic
acid which is present in 1 gram of fat. It is used to
measure the free fatty acid (FFA) that are present in
the fat or oil.Saponification value is defined as the
amount of KOH (potassium hydroxide) required in
milligrams to to turn 1 gram of fat into soap. It
depends on the nature of the fatty acids that are
contained in the fat. The iodine value is the number
which expresses in grams the quantity of Iodine,
which isabsorbed by 100 g of the substance. The
iodine value indicates the degree of unsaturation of a
fat or oil. It is defined as the number of grams of
iodine absorbed by 100 g of fat.