Functions of Cholesterol:

• Cholesterol is required to build and maintain membranes.

• It modulates membrane fluidity over the range of
physiological temperatures.
• Cholesterol serves as a precursor for the biosynthesis of
steroids hormones, bile acids, and vitamin D.
• Cholesterol is the principal sterol synthesized by animals.
• Recent studies show that vitamin D is a potent
antioxidant, helping to detox the body and protect arteries
too.
Functions of Cholesterol:

• Cholesterol is an important precursor molecule for the

synthesis of Steroid hormones Including the adrenal gland
hormones
cortisol
aldosterone
• As well as the sex hormones
progesterone
estrogens
testosterone
Identification Tests for Cholesterol

• Modified Abell-Kendall method:

• Cholesterol is hydrolyzed with alcoholic KOH
• Unesterified cholesterol is extracted with petroleum jelly
• measured by using Libbermann-Burchardt reaction

Cholesterol + sulphuric acid + acetic anhydride

↓
bluish green colour
Synthesis of Cholesterol

• Robinson synthesis:
• The starting compound for
this synthesis is 1,6-
dihydroxy naphthalene.
• After 20 steps it is
converted into
androsterone
• Androsterone is then
converted into
androstenedione by
oxidation.

• androstenedione is
converted into
dehydroepiandrosterone
• Dehydroepiandrosterone
is converted pregnenolone
by multiple steps.

• Pregnenolone leads to a
compound known as
allopregnenolone
• The addition of acetate in
allopregnenolone leads to
cholestanol.
• The oxidation of
cholestanol gives ketone.
• In the next step
bromination of ketone
produces a bromo-ketone.
• After the formation of
enone by the elimination of
bromo-ketone, the final
product is cholesterol.
Over all reaction:
 Desalting and the Choice of Buffers
• Desalting is probably the most important step in
the separation of water soluble compounds.
• Strongly hydrophilic compounds are routinely
analyzed on HPLC.
• Preparative separation of water soluble compounds
is hindered by the inorganic salts.
• This step is often given up by organic chemists.
 Desalting and the Choice of Buffers
• If the compounds have some lipophilicity, the
standard method for desalting is the use of reverse
phase columns such as C18 silica gel or other
organic polymers like XAD-2, XAD-4 and XAD-7.
• Usually, the aqueous solution is passed through
the column and salts are washed out with water.
• Another commonly used method for desalting is the
use of size exclusion cloumn like Sephadex G-10
and Bio-Gel P-2. e.g., purification of paralytic…….
Desalting and the Choice of Buffers
• Purification of paralytic shellfish toxins such as
saxitoxins and gonyautoxins was achieved by taking
advantage of their specific adsorption on Bio-Gel P-2
of Sephadex G-10.
• In most cases, extraction of water soluble
compounds necessitate the use of buffer solutions.
• The success for the use of buffer solutions is to find
the appropriate pH of the buffer solutions.
• Isolation of buffer salts from compounds is
problematic
Desalting and the Choice of Buffers
• Buffer solutions have their own disadvantages.
e.g., buffer solutions containing pyridine have very
unpleasant smell.
• Ammonium acetate and ammonium carbonate
buffer can be removed by evaporation or
lyophilization, but it is not an easy task. The first
lyophilization leaves sticky mixture of ammonium
acetate or carbonate solutes.
• Sometimes this process has to repeat.
 Structure Elucidation of Alkaloids
Von Braun Degradation Method
• The von Braun reaction is a reaction in which a tertiary
amine reacts with cyanogen bromide to an organocyana
amide. For example

Mechanism
The reaction mechanism consists of two nucleophilic
substitutions ; the amine is the first nucleophile displacing the
bromine atom which then acts as the second nucleophile. For
example
19
 Structure Elucidation of Alkaloids
Von Braun Degradation Method

• First, the amine reacts with the cyanogen bromide to form a

cyano group. This results in a quaternary ammonium salt,
which in the next step reacts by splitting off bromomethane to
give the dimethylcyanamide. This is a second-order nucleophilic
substitution (SN2)

20
 Structure Elucidation of Alkaloids
Von Braun Degradation Method
• The von Braun amide degradation is the chemical reaction of a
mono substituted amide with phosphorous pentachloride or
thionyl chloride to give a nitrile and an organo halide. It is
named after Julius Jacob von Braun, who first reported the
reaction.

21
 Structure Elucidation of Alkaloids
Von Braun Degradation Method
• The von Braun reaction is a reaction in which a tertiary
amine reacts with cyanogen bromide to an organocyana
amide. For example

Mechanism
The reaction mechanism consists of two nucleophilic
substitutions ; the amine is the first nucleophile displacing the
bromine atom which then acts as the second nucleophile. For
example
19
 Structure Elucidation of Alkaloids
Von Braun Degradation Method

• First, the amine reacts with the cyanogen bromide to form a

cyano group. This results in a quaternary ammonium salt,
which in the next step reacts by splitting off bromomethane to
give the dimethylcyanamide. This is a second-order nucleophilic
substitution (SN2)

20
 Structure Elucidation of Alkaloids
Von Braun Degradation Method
• The von Braun amide degradation is the chemical reaction of a
mono substituted amide with phosphorous pentachloride or
thionyl chloride to give a nitrile and an organo halide. It is
named after Julius Jacob von Braun, who first reported the
reaction.

21
 SYNTHESIS OF NICOTINE
Spath et al. describe the synthesis of nicotine.
 Structure Elucidation of Alkaloids
Van Braun’s Degradation Method
• There are two types of von Braun degradations.
• In the first method, the tertiary amine that contains at least one
alkyl substituent, is treated with cyanogen bromide. This results in
the cleavage of an alkyl nitrogen bond to give an alkyl bromide and
a substituted cyanamide.

• In the second von Braun’s method, secondary cyclic amine is

reacted with benzoyl chloride in the presence of NaOH to afford
benzoyl derivative, which on treatment with phosphorous
tribromide-bromine followed by distillation under reduced
pressure yields α, ω-dihalo compound with the elimination of
benzonitrile, for example piperidine in this degradation yields 1,5-
dibromopentane and benzonitrile. 16
Structure Elucidation of Alkaloids
Van Braun’s Degradation Method

17