Dha.

Ilangeswaran 1
 Introduction
A secondary alcohols with 27 to 29 carbon atoms of
animal or plant origin.
Crystalline solids with m.p. range of 100 – 200oC.
The terms ‘stero’ – means solid & ‘ol’ – alcohol.
Number of compounds such as vitamin D, bile acids, sex
hormones, adrenal cortex hormones are belonging to
steroids.
The structure of steroids are based on 1,2-
cyclopentenophenantherene skeleton.
Sterols occur in animal and plant oils & fats. They occur
as free or an ester of the higher fatty acids.
They are isolated from unsaponifiable portion of oils &
fats.
Dha. Ilangeswaran 2
 3 3'
4 2

5 2'
6 1 1'

7 10
8 9
1,2-Cyclopentenophenanthrene

Any compound which gives Diels hydrocarbon with Se

distillation is defined as steroid. When steroids are distilled at
420oC yields mainly chrysene and a little picene.
2
1 3
CH3
12 1 14
11 2 13 4
10 12
3 5
9 11
4 6
8 5 7
10
7 6 9 8
Diels Hydrocarbon Chrysene Picene
Dha. Ilangeswaran 3
 Classification
Sterols

Zoosterols Phytosterols Mycosterols

(Animal Sources) (Plant Sources) (Yeast & Fungi Sources)
CH3
21 22
H3C 20 23 H3C CH3
26
18CH3 CH3 CH3
17 CH3
19 12 24 25
11 13 CH3
CH3 16 27 CH3
1
2 9 14
10 8 15
H
HO 3 5 7
6 HO
4 Ergosterol (mycosterol)
Cholesterol (zoosterol) C28H44O m.p. 165oC
C27H46O m.p. 149oC
29
28 CH3
21 22
H3C 20 24
CH3 CH3
23 25 26
H3C
CH3 27

HO
Stigmasterol (phytosterol)
C29H48O

Dha. Ilangeswaran 4
 Constitution of Cholesterol
1. Molecular formula: C27H46O
2. Presence of double bond and hydroxyl group:
a. The conversion of cholesterol into cholestanol shows
the presence of double bond.

H2 - Pt
C27H46O C27H48O
Cholesterol Cholestanol
b. The oxidation of cholestanol with chromic acid into
cholestanone shows the presence of a secondary
alcoholic group in cholesterol.
CrO3
C27H48O C27H46O
Cholestanol Cholestanone
 c. The Clemmenson’s reduction of cholestanone
yields a saturated hydrocarbon called cholestane.
Zn/Hg - HCl
C27H46O C27H48
Cholestanone Cholestane

3. Presence of a ring system: The molecular formula

of cholestane corresponds to the general formula
CnH2n-6, of a tetracyclic system. On distillation with
Se at 360oC, cholesterol yields Diel’s hydrocarbon.
CH3
2 3'
Se 2'
C27H46O 1 1' +
360 °C
Cholesterol
3'-Methy-1,2-cyclopentenophenanthrene Chrysene
(Diel's hydrocarbon)

Dha. Ilangeswaran 6
 The above reaction shows the presence of a
cyclopentenophenanthrene nucleus in cholesterol and thus
cholesterol is a steroid.
The various rings in the cholesterol opened in
different conditions to give various dicarboxylic acids.
The relative positions of two carboxylic groups with
respect to each other were determined with the help of
Blanc’s rule.
Blanc’s rule: It states that on heating with acetic
anhydride, 1,5-dicarboxylic acids form cyclic anhydrides
and 1,6-dicarboxylic acids form cyclopentanone with the
elimination of carbon dioxide.

Dha. Ilangeswaran 7
Ring A: Cholesterol and cholic acid (which is structurally
related to the former) were converted into the dicarboxylic
acid A which on heating with acetic anhydride gave a
cyclopentanone. Therefore the ring A in cholesterol may be
six membered on the basis of Blanc’s rule.
R
R

C D (CH3CO)2O
C D
HOOC
B O
B
HOOC
Dicarboxyllic acid A

Dha. Ilangeswaran 8
 Ring B: The tricarboxylic acid derived from cholesterol on
heating with acetic anhydride gave cyclopentanone derivative
and carbon dioxide. Therefore on the basis of Blanc’s rule the
ring B in cholesterol may be a six membered.

R R

(CH3CO)2O
C D D
HOOC C

COOH HOOC
O
COOH
Tricarboxyllic acid (B)

Dha. Ilangeswaran 9
Ring C: Deoxycholic acid (structurally related to cholesterol &
cholic acid) was converted into a dicarboxylic acid which gave a
cyclic anhydride when subjected to Blanc’s reaction condition.
Hence the ring C was assumed to be a five membered. On this basis
Windaus and Wieland proposed the following structure to
cholesterol.

CH3
CH3
D
B C2H5
CH - (CH2)3 - CH (CH3)2
A C

OH

Dha. Ilangeswaran 10
Later in 1930, Wieland et al proved that there was no ethyl
group at C10 and proposed that steroids contain chrysene
nucleus and cyclopentenophenanthrene structure. Here Blanc’s
rule failed. If we use the correct structure of cholesterol, the
cyclisation reaction results in the formation of seven
membered cyclic anhydride. Thus ring C also six membered.
R
R
HOOC O
O O
HOOC D D
(CH3CO)2O

A B A B

Dicarboxyllic acid (C)

Dha. Ilangeswaran 11
Ring D: Cholestane was converted into etiobilianic acid, D
which gave a cyclic anhydride. Hence according to Blanc’s
rule the ring D may be a five membered one.
O

COOH
O
C (CH3CO)2O C
COOH
A O
A B B

Etiobilianic acid (D) Anhydride

Dha. Ilangeswaran 12
 Position of the Hydroxyl Group & Double Bond
The positions of hydroxyl group and double bond were fixed
with the help of following reaction.
HNO3 300 °C
C27H46O C27H46O4 C26H44O
Cholestanone Dicarboxyllic acid Ketone
Here thea dicarboxylic acid and cholestanone contain the same
b c

number of carbon atoms. Hence cholestanone must contain the

keto group in the ring. Further pyrolysis of dicarboxylic acid
gave a ketone, c. According to Blanc’s rule compound b may be
1,6 or 1,7-dicarboxylic acid. Already it was established that
cholesterol contains 3 six membered & 1 five membered ring.
Here the dicarboxylic acid may be produced by the opening of
ring A or B or C. Hence –OH group may be present any one of
the above three rings.
 When cholestanone is oxidized, we get two isomers of
dicarboxylic acids. This will be possible only if keto group in
cholestanone is flanked on either side with methylene group
(-CH2COCH2-). This arrangement is possible only in ring A.
Hence the hydroxyl group in cholesterol must be present in ring
A.
Now consider the following set of reactions.
H2O2 CrO3 (i) - H2O
C27H46O C27H48O3 C27H44O3 C H O
27 44 2
CH3COOH (ii) Zn - CH3COOH
Cholesterol Cholestanetriol Hydroxycholestanedione Cholestanedione
I II III IV
CrO 3

C27H44O8
Tetracarboxylic acid
V

Dha. Ilangeswaran 14
 In the above reactions, the conversion of cholestane triol(II)
to dione(III) showed that in compound II, two –OH groups are 2o and
the third one which resists oxidation may be 3o.
The formation of IV from III without loss of carbon atoms
showed that the two keto groups may be present in different rings.
Hence in cholesterol the –OH group and the double bond may be
present in different rings.
Compound IV, cholestane dione forms a pyridazine derivative
with hydrazine. This is a characteristic property of -diketone. Hence
compound, IV must be a -diketone.
The position of hydroxyl group is already fixed with ring A.
All the above reactions can be explained well if we place –OH
group at C3 position of ring A and the double bond between C5 and C6
of ring B.

Dha. Ilangeswaran 15
 B H2O2 CrO3
A A B A B
HO
CH3COOH O
HO
Cholesterol OH OH
I OH O
Hydroxycholestanedione
Cholestanetriol
III
II
O
H 2
OH
( i) - C O
CH
3
HOOC CrO3 -
A B i) Zn
(i
COOH O
HOOC H
COOH O NH2NH2
Cholestanedione
Tetracarboxylic acid IV
V

N
H

Pyridazine derivative N

Dha. Ilangeswaran 16
 The above fact is further supported by the following reactions

CuO KMnO4
290 °C HO
+ CO2
HO O O
Cholesterol Cholestenone
VII O Keto acid VIII
I

Cholesterol on heating with copper oxide at 290oC gave

cholestenone, which on oxidation with permanganate yielded
keto acid with a loss of carbon atom. The formation of keto acid
revealed the presence of keto group and a = bond in the same
ring of cholestenone. These results can be explained if it is
assumed that the double bond in cholesterol migrates during the
formation of cholestenone.
Dha. Ilangeswaran 17
 The position of hydroxyl group at C3 is finally proved
by Kon et al. The formation of 3’,7-dimethyl
cyclopentenophenanthrene from cholesterol by the following
steps is possible only if –OH group is considered at position
C3 .
H2 / Pt CrO3

HO HO O
H H
Cholesterol
I CH3MgBr
CH3

3'
Se
350 °C H3C
HO
H3C 7 H

3',7-Dimethylcyclopentenophenanthrene

Thus we concluded that cholesterol contains –OH group

at position C3 and a double bond between C5 to C6.
Dha. Ilangeswaran 18
 Nature of the Side Chain
The cholesteryl acetate derived from cholesterol
on oxidation with CrO3 forms a ketone (steam volatile) &
acetate of hydroxy acetone (non-steam volatile). The ketone was
found to be isohexyl methyl ketone, which may be the side
chain of cholesterol. The point of attachment of side chain may
be at the carbon of the keto group, i.e. at C17 – position.
O

H3C CH3
+
O CH3
AcO
Isohexylmethyl ketone
 The nature of side chain and the linkage have been
studied by Barbier – Wieland. The B-W degradation offers a
method of getting a lower acid with one carbon atom less as
described below.

CH3OH 2C6H5MgBr
RCH2COOH RCH2COOCH3 RCH2C(OH)(C6H5)2
HCl
- H2O

CrO3
RCOOH + O=C(C6H5)2 RCH=C(C6H5)2

Dha. Ilangeswaran 20
 Cholesterol was first converted into 5 -cholestane (a
stereoisomer of cholestane). The nucleus of 5 -cholestane is
represented as Ar and side chain as Cn, then the degreadations
can be expressed as follows.

5 -Cholestane (or coprostane) CrO3 B-W

CH3COCH3 + Cholanic acid
Ar-Cn Ar-Cn-3

B-W B-W
(C6H5)2CO + Norcholanic acid (C6H5)2CO + Bisnorcholanic acid
Ar-Cn-4 Ar-Cn-5

CrO3 Etianic acid

(C6H5)2CO + Etiocholylmethyl ketone
Ar-Cn-6 Ar-Cn-7

Dha. Ilangeswaran 21
 The formation of acetone from 5 -cholestane shows
that side chain terminates in an isopropyl group. The
conversion of bisnorcholanic acid into a ketone reveals the
presence of an alkyl group on the -carbon in bisnorcholanic
acid. As etiocholyl methyl ketone is oxidised to etianic acid
with a loss of one carbon atom, the ketone may be a methyl
ketone, i.e., there is a methyl group in -carbon of bisn
orcholanic acid.
When etianic acid is subjected to one more B – W
degradations, a ketone (etiocholanone) is obtained which on
oxidation using nitric acid yields dicarboxylic acid
(etiobilianic acid) without loss of carbon atoms. Hence
etiocholanone must be a cyclic ketone.
These reactions shows the presence of eight C atoms
in side chain.
Dha. Ilangeswaran 22
 The above degradations can only be explained if the
side chain has the following structure.
CH3
CH3

Ar - CH - CH2 - CH2 - CH2 - CH

CH3
Nucleus Side chain

Position of Side Chain: Etiobilianic acid derived from

etiocholanone forms an anhydride on treatment with acetic
anhydride. Acc. to Blanc’s rule etiocholanone may be a 5
membered ketone. So the side chain may be attached to the 5
membered ring D. The formation of Diel’s hydrocarbon from
cholesterol suggests that the side chain is at C17 as
dehydrogenation degrades a side chain to methyl group.

Dha. Ilangeswaran 23
 Further 5 -cholanic acid is formed from 5 -
cholestane on oxidation. Deoxycholic acid also on oxidation
followed by Clemmenson’s reduction yields 5 -cholanic acid.
the side chain in cholesterol & deoxycholic acid may be
present in the same position. Now the nature & position of
side chain is known the conversion of 5 -cholestane to
etiobilianic acid can be expressed as follows.

Dha. Ilangeswaran 24
 H3C
H3C
CH3 CH3
CH3 COOH
O
+
CrO3 CH3
H3C
5 -Cholestane COOH
5 -Cholanic acid B-W

COOH
H3C O H3C

B-W B-W
Nor-5 -Cholanic acid

Etiocholylmethyl ketone Bisnor-5 -Cholanic acid

CrO3

COOH O H3C
COOH
COOH
B-W HNO3

Etianic acid Etiocholanone Etiobilianic acid

Dha. Ilangeswaran 25
Position of Two Angular Methyl Groups
When anhydride of etiobilianic acid is distilled with Se,
1,2-dimethylphenanthrene is obtained. It shows the presence of
phenanthrene nucleus and an angular methyl group at C13 of
cholesterol.
The cyclopentenophenanthrene nucleus of cholesterol
accounts for 17 carbon atoms & the side chain for 8 C atoms.
This accounts for 25 C atoms and the remaining 2 are assumed
to be angular methyl groups.
When the position of –OH group & = bond are
determined one of the compounds formed was keto acid, VIII.
This on subjected to Clemmenson’s reduction followed by two
B-W degradations, we get an acid which is difficult to estrify
and gives CO on warming. This clearly shows that –COOH
group must be linked to a tertiary C atom & side chain must be
of the following type.
Dha. Ilangeswaran 26
 C C
2 (B - W)
C - C - C - C - COOH C - C - COOH

C C
The reactions are shown below

HOOC
Zn - Hg 2 (B - W)
COOH COOH
O
HCl
Keto acid Acid group with
VIII 3o carbon atom
This shows an alkyl group at C10-position.
To determine the position of 2nd methyl group, consider
the Se dehydrogenation of cholesterol to yield chrysene & Diel’s
hydrocarbon. The formation of chrysene can be explained if
there is an angular methyl group at C13 of cholesterol, which
enter into 5 membered ring during Se dehydrogenation to give
4th six membered ring of chrysene.
Dha. Ilangeswaran 27
 The positions of 2 angular methyl groups at C10 &
C13 are supported by the following reactions.
OH O O
COOH COOH
COOH

CrO3 HNO3
COOH
HO HO HOOC
Deoxycholic acid Dehydrodeoxycholic acid Deoxybillianic acid

O
O
COOH COOH
COOH
HNO3 KMnO4

COOH COOH
COOH
A O
Diketo-dicarboxylic acid O
+
HOOC COOH O COOH
COOH
HNO3 HOOC
HOOC
H HOOC
HOOC H
COOH
B C D

Dha. Ilangeswaran 28
 The compound A was found to be butane-2,2,4-tricarboxylic
acid. This shows an angular methyl group at C10. Compound B is a
tetracarboxylic acid with a cyclopentane ring and a side chain having
–C-Me group. D is a tricarboxylic acid where –COOH is linked to
tertiary carbon. This proves an angular methyl group at C13.

The conversion of etiobilianic acid into 1,2-dimethylphen

anthrene shows angular methyl group at C13. If it is at C14 the
product should be 1-methylphenanthrene.
CH3 CH3
COOH O
CH3
CH3 O
COOH CH3 Se
CH3
O

Etiobilianic acid Anhydride 1,2-Dimethylphenanthrene

Dha. Ilangeswaran 29
 Androsterone
Molecular formula: C19H30O2
Melting point: 183oC, [ ]D = +94o
It was first isolated by Butenandt et al. in 1931. Androsterone
behaves as a saturated compound.
As it forms mono-ester, one oxygen atom is present as a
hydroxyl group. The 2nd O atom was shown to be oxo, since
androsterone forms an oxime.
The parent hydrocarbon of androsterone is C19H32, and since
this corresponds to the general formula CnH2n-6, it may be
tetracyclic.
D.B.E. of C19H30O2 = (a+1) - (b-c)/2 = (19+1) – (30-2)/2 = 5;
1 double bond due to C=O, & 4 rings.
Dha. Ilangeswaran 30
 The above facts suggest that androsterone may conatain
a steroid nucleus. Butenandt proposed a structure which was
proved to be correct by Ruzicka as follows.
Ruzicka oxidized 5 -cholestanyl-3 -acetate with
chromic oxide in acetic acid to epiandrosterone, which is a
hydroxyketone.
H3C
O
CH3 CH3 CH3

CH3
CH3 H CH3 H
(i) CrO3
H (ii) Hydrolysis H
AcO HO
H H
5 -Cholestanyl-3 -acetate Epiandrosterone

Dha. Ilangeswaran 31
 But the oxidation of 5 -cholestanyl-3 -acetate only
yields androsterone.
H3C
O
CH3 CH3 CH3

CH3 CH3 H
CH3 H
(i) CrO3
H H
(ii) Hydrolysis
AcO HO
H H

5 -Cholestanyl-3 -acetate Androsterone

Thus the configuration of the –OH group at C3 is and

not as Butenandt suggested. Epiandrosterone, m.p. 174oC,
[ ]D + 88o, has about 1/8th of the activity of androsterone. In
1955 Sondheimer et al. converted epiandrosterone into
androsterone as follows.
Dha. Ilangeswaran 32
 O
O O
CH3
CH3 CH3

CH3 H
CH3 H CH3 H
AcONa
H
+
H AcOH-Ac2O H

TsO H AcO
H H
54% 39%
Epiandrosterone-p-toluenesulphonate

H2O2 NaOH
O
O
CH3
CH3
O
CH3 H
CH3 H

H
H
HO
H Androsterone
H

PhCO3H Aq. AcOH

O
O
CH3
CH3
O
O
CH3 H
CH3 H
LiAlH4
O H
H

H HO
H

Dha. Ilangeswaran 33
 But a conventional preparation of androsterone starts
from dehydroepiandrosterone.
O O
CH3 CH3
O
CH3 H CH3 H
H2O2 Oppenauer
H H TsOH / PhH H H oxidation
HO HO

Dehydroepiandrosterone
O
CH3
O
CH3
O CH3 H H2O2
CH3 H (i) B2H6
H H
TsOH / PhH
H H (ii) Ac2O
O H

O
O CH3
CH3
O (i) B2H6
CH3 H
CH3 H (ii) H2O2/OH
H H
H H (iii) acid
HO
H Androsterone
H
Dha. Ilangeswaran 34
 A total synthesis of androsterone has been carried out
by Woodward et al. using the following ester, which is used
in the synthesis of cholesterol. COOMe
CH3

CH3 H

H H
O
H
Soon after the discovery of androsterone, Butenandt et
al. isolated the following hormones from male urine.
OH
O CH3
CH3

CH3 H
CH3 H

H H
H H
O
HO
H
5 -androsterone Testosterone
m.p. 151oC, [ ]D = +105o
Dha. Ilangeswaran 35