Steroids and Hormones

These are the compounds, which are widely distributed in animals and plants and have
1,2-cyclopenteno-perhydrophenanthrene nucleuses in their structure. The steroids include a variety of
compounds eg sterols including vitamin D, bile acids, sex hormones, certain adrenal cortical
hormones, cardiac and toad poison, sapogenins, some alkaloids, some carcigonic hydrocarbons etc.
On selenium dehydrogenation at 360oC all the steroids yield Diel’s hydrocarbon
(3 -methyl-1,2-cyclopentenophenanthrene), where as at 420oC chrysene is the main product with
/

some picene. 3 3 /
CH3
4 2/
2 1
5 1 1/
6
2

7 10
Chrysene 8 9
1,2-cyclopentenoperhydro-
Picene Diel's hydrocarbon
phenanthrene

1,2-
Diel’s Hydrocarbon (m.p. 126-127oC): cyclopentenoperhydro
phenanthrene
Its molecular formula is C18H16. Oxidation reaction, X-ray crystal analysis and
absorption spectrum measurement showed that the Diel’s hydrocarbon is 3 /-methyl-1,2-
cyclopentenophenanthrene. The structure of the compound is proved by its synthesis.
CH3
CH2MgBr CH3 OH
CH2
O
+
CH3
CH3
2-(1-naphthyl)-
ethyl- 2,5-dimethyl
magnesium cyclopentanone P2O4 at 140o
bromide
distilled under
reduced pressure

H CH3
CH3

CH3 CH3

Se
CH3

Diel's hydrocarbon
(3/-methyl-1,2-cyclopentenophenanthrene)
 20, etc.
Cholesterol, notorious as the R 18
20, etc.
11 13
substance deposited on the walls of 18 17
19 12 R
12 2 C D 17
arteries and as the chief constituent of 1 19
11
C
13
D 16 A
1 10
B 9
8
15
14
gallstones, is the kind of alcohol called a 2 9
8
14
15
3
4
5
6
7
16

10
sterol that belongs to the class of 3 A B
7
5
compounds called steroids. The general 4 6
A steroid
structural formula of steroids are shown
in the figure. The rings are (generally aliphatic. Lines like the vertical ones attached to the
10-and13-positions represent 21
26
CH3 CH2
angular methyl groups. 21
23
26
25
For example, R for Commonly, in cholesterol, R= CH CH2 CH2 CH2 CH or 20
20 22 23 24 25 22 24 27

cholesterol shown in the CH2

27

figure. Stereochemistry is indicated

19
by solid lines (-bonds, coming out 1 H3C
2
H3C
9
of the plane of the paper) and broken 2
19
A 5 10 B A
1 10
B 9
8

3
lines (-bonds, going behind the C 6 7 3
4
5
6
7
H 4
H
plane of the paper). Thus in the 2
1
19
9
trans Fusion H
figure showing –H and –OH at the 3
A 5 10 B H
CH3
5- and 6-positions are cis to each HO 4 H 6 7
H
1 3 C 10
B
other, but trans to the 3-OH and to H
2 9
H
19 10
the angular methyl at the 10-position. A 3, 6-diol
3
A 5 B 5

6 7 A
Fusion of the rings to each other can 4 H
be cis or trans, thus increasing the cis Fusion
complications of the stereochemistry.
Finally, in any rigid cyclic system like this, con formational effects are marked, and often
completely control the course of reaction. Steroids include sex hormones and adrenal cortical
hormones, cardiac glycosides and bile acids. Because of their biological importance and,
undoubtedly, because of the fascinating complexity of the chemistry the study of steroids has been,
and is now, one of the most active areas of organic chemical research. Cholesterol, despite its bad
popular image, is an essential constituent of all cells, and is an intermediate in the biogenesis of the
other steroids.
Nomenclature of steroids: Steroids are numbered as shown in the formula. When some of the
carbon atoms in the formula are missing, the numbering of the remainder remains unchanged.
Solid lines (preferably
H
thickened) denote groups above 21 22 29
C
21 28
26 C D
the plane of the nucleus 18 17
20
24 25 27 H H
D H H
12
(-configuration), and dotted 11
C
13
H A B
H H
1 19 D 16 A H
or broken lines denote groups 2 9 14
B
10
8 15 H
below the plane 3 A B H
7
5
(-configuration). If the 4 6

configuration of the substituent A steroid 5 (  or )-gonane 5 (  or )-oestrane

is unknown, its bond to the nucleus is drawn as a wavy line and this is indicated in the name.
Wherever possible, the name of the steroid should be specifying stereochemical configuration.
When a methylene group is missing from the side chain, this is indicated by the prefix ‘nor’
preceded by the number of the carbon atom that has disappeared. When a ring has been contracted or
enlarged, this is indicated by prefixes ‘nor’ and ‘homo’ respectively, preceded by a small capital
letter indicated the ring affected. The prefix ‘nor’ is also used to indicate the loss of an angular
methyl group, and in this case is preceded by the number designating that methyl group: 18-nor and
19-nor. When ring-fission has occurred with addition of a hydrogen atom to each new terminal group,

2
this is indicated by the numbers showing the position of the bond broken, followed by the prefix
‘seco’. The prefix ‘cyclo’, preceded by the numbers of the positions concerned, is used to indicate a
three-membered ring. Some examples of these rules are shown in the figure.
Et

C C D C D
D H H
H H

H A H H A H H
A H
B B B
H H H
23-nor-5cholane A-nor-5-androstane B-homo-5pregnane

C D C D
C H H
D
H
HOOC A H H H H
B A B
H H
HOOC
A B H
H
3, 4-seco-5cholane 2, 3-seco-5-androstane-2,3-dioic acid 3-cyclocholestane
Compounds derived from 5-cholestane belongs to the allo-series, the prefix ‘allo’ being
reserved to indicate this configuration (ie, 5). Compounds derived from 5-cholestane (coprostane)
belong to the normal-series. It is not customary to prefix compounds of the latter series by the word
‘normal’; eg, cholanic acid can be derived from 5-cholestane (coprostane).
Spectral properties of steroids: Infrared spectroscopy, based on the knowledge gained from the
study of steroids of known structures, is extremely valuable in elucidating the structures of unknown
steroids. The more important groups are OH, C=O, C=C, C=C–C=O, COOH, COOR, and these have
their maxima in the ranges given. A very important feature of steroid infrared spectra, however, is the
dependence of the absorption maximum of the keto group on its position in the nucleus, and also, in
some cases, according to whether the hydrogen atom at the ring junction is  or .
The absorption maxima have been compiled in the following way: For saturated ketones
(C=O str. Cm-1): 3-CO (5 and 5), 1719-1712; 4-CO (5), 1712; 4-CO (5), 1713; 6-CO (5),
1714-1712; 6-CO (5), 1708-1706; 11-CO (5), 1710-1704. When the chromophore is the
, -unsubstituted carbonyl group, both the C=O (str.) and C=C (str.) have the maxima in regions
depending on the position of the chromophore in the molecule, eg, ∆1-3-ketone: C=O, 1684-1680,
and C=C, 1609-1604; ∆4-3-ketone: C=O, 1681-1677, and C=C, 1619-1615. It was also found that if
the nucleus contained two keto groups, each showed its own maximum (or one very close to it) in
some cases, but was different in other cases. An example of the first type is the 3, 17-diketo
compound (1719, 1745 cm-1), and an example of the second type is the 11, 17-diketo compound
(1713, 1751 cm-1). In the latter case, the two keto groups are sufficiently close to each other to allow
vibrational interaction, which has been called a vicinal effect.
The ultraviolet absorption spectra of steroids are characteristic of the functional groups present,
and the observed maxima are in good agreement with those calculated from Woodward’ rules.
The following examples illustrate these rules, all the compounds being steroid derivatives; only the
part of the molecule concerned is shown in the figure. In example (I), the diene is homoannular, there
are three substituents, and there is one exocyclic double bond. The 4,5-double bond is part of the
homoannular diene system in ring A, but is exocyclic with respect to ring B, and is therefore 5 nm
3
must be added. Thus the calculated
value is 253 + 3 x 5 + 5 = 273 nm.
In an example (II) the basic diene
system has been taken to be the
homoannular one in ring B. HO O
There are then two double bonds (I) (II) (III)
extending this conjugation, and the obs. 275 nm obs.355 nm obs.244 nm
double bond in ring A is exocyclic to calc.273 nm calc.343 nm calc.244nm
ring B. Thus the calculated value is
253 + 2 x 30 + 5 x 5 + 5 = 353 nm.
The rules apply to three double
bonds in conjugation, but may break
O O
down for four, and definitely cannot
be applied to five or more double (IV) (V)
bonds in conjugation. In compound obs.230nm obs.241 nm
(III) there is a crossed conjugation. calc.227nm calc.244nm
In such cases the calculation is made for the linear system, which gives the higher value for max.
Thus, the linear system using the 1,2-double bond gives 227 nm (this is equivalent to (IV), but for the
system with the 4,5-double bond, the value is 244 nm (this is equivalent to (V). Hence the calculated
value is taken as 244 nm.
NMR spectroscopy has been used in the structural studies of steroid, but complete analysis of
such complex molecules is extremely difficult. Steroids appear to have a finger print region, which is
characteristic of the CH and CH2 protons in the nucleus. Also, the proton in the =CH– has a definite
-value, and so this group can usually be readily detected, and the number of these groups present can
be estimated from the area of the proton peak. Furthermore, from the table compiled from spectra of
known compounds it may be possible to determine the position of a double bond in unknown
steroids. It was found that the protons of the angular methyl groups at C-10 and C-13 have their own
characteristic chemical shifts, the actual shift depending on the presence of various functional groups
such as C=C, C=O. Thus a 3-keto group causes a shift of 0.15 ppm in the C19-methyl line, but has no
effect on the position of the C18-methyl line. Hence it may be possible to deduce both the nature and
position of a functional group in the molecule. It was also found that the methyl protons of an
acetoxyl group have a characteristic -value depending on its position in the nucleus. In this way, it is
possible to determine the position of a hydroxyl group in the molecule.
Mass spectrometry is very useful for structural analysis of steroids. Steroids usually give an
abundant molecular ion and so it is easy to determine the molecular weight and molecular formula.
Four common peaks usually observed are: (a) [M-R]+, where R is the side-chain; (b) [M-(R + 42)]+,
where mass 42 is C3H6 ; (c) [M-15]+, due to loss of an angular methyl group; (d) [M-(R + 42 +15)]+.
Because of this general fragmentation pattern, it is possible to detect the presence of a steroid
nucleus. Steroid alcohols usually show the presence of a peak at m/e [M-18]+ due to the loss of water,
and a peak at [M – (18 + 15)]+ due to loss of both water and an angular methyl group. On the other
hand, the fragmentation pattern of ketones depends on the position of the keto group, and it appears
that the positive charge always resides with the oxygen-containing fragment. However, it has been
found that ethylene ketals are particularly useful for the purpose of directing fragmentation in a
predictable manner. Since the 3-hydroxyl group and 3-keto groups are very common in steroids, the
cracking pattern of 3-ethylene ketals has been studies and three ions that have been observed in
greatest amount are shown in the figure (VI, VII and VIII). In a steroid with a 3-keto group and a
5,6-double bond, fragment (VI) shows an intense peak and the peaks of (VII) and (VIII) are
extremely weak. On the other hand, if the steroid contains a 7-keto group, the predominant peak is
(VII). It can thus be seen that it is possible to locate a 3- or 7- keto group and a 5,6 double bond in an

4
unknown steroid.
+ +
However, there are O O O O O O
some complications, O
since a 17-ethylene O
ketal also gives (VI). H
Another interesting
application of mass (VI) (VII) (VIII)
spectrometry is the
location of an
ethylernic double O (VII)
(VI)
bond in a steroid O
molecule. The double O O
H
bond is first converted
into the epoxide, which, on treatment with dimethylamine, forms the dimethylamino-alcohol.
This, on electron impact, is split into fragments, and the m/e value of the nitrogen-containing
fragment indicates the position of the double bond.
R R R

-e
(-H2O)

HO
NMe2 NMe2
Cholest-5-ene

NMe2

m/e 276
 Sterols:

These are found in animal and plant oil and fats. These are crystalline compounds containing a
secondary alcoholic group. So they differ from common alcohols in being solid and due to this reason
they are known as sterols (solid alcohols). Roughly they may be divided into three groups: zoosterols
(which are obtained from animals, eg., cholesterol, cholestanol and coprostanol); phytoserols
(obtained from plants, eg., stigmasterol); and mycosterols (from yeast and fungi, eg., ergosterol,
vitamin D). However it is not a rigid classification because some of the sterols are obtained from
more than one source, eg., ergosterol.
i) Cholesterol: It is an animal sterol and occur either free or as fatty esters. The main sources are
brain, spinal cord, gallstone and fish liver oil. It forms about one-sixth of the dry weight of nerve and
brain tissue and is obtained commercially by the extraction of cattle spinal cords. As it was first of all
isolated from human gallstone deposited in the bile duct, it is known as cholesterol (chole means bile;
sterol). The human body not only can synthesize cholesterol but can also absorb it from food through
the intestine into the bloodstream. Too high a concentration of cholesterol in the blood can lead to its

5
precipitation in the circulatory vessels, resulting in high blood pressure and arteriosclerosis.
It is white crystalline optically active solid with m.p. 149 o C and specific rotation (+) 39o.
Cholesterol (and other sterols) gives the following color reactions:
i)The Salkowski reaction: A solution of cholesterol in chloroform, when treated with
concentrated sulphuric acid, develops a red color in the chloform layer.
ii)The Libermann-Burchard reaction: A solution of cholesterol in chloroform, when treated
with concentrated sulphuric acid and acetic anhydride gives a greenish color.
iii)Ethanolic solution of cholesterol, when treated with a ethanolic solution of digitonin
(a saponin), gives a white precipited of cholesterol digitonide. The molecular complex of cholesterol
digitonide when dissolved in pyridine, dissociates to its components. The individual components may
be separated by ether when cholesterol remains in solution while the digitonin is precipitated.
Alternatively, the digitonide is dissolved in dimethyl sulphoxide and the solution is heated on a steam
bath. On cooling, only the sterol is precipitated. Actually, digitonide formation is used for the
estimation of cholesterol.
Constitution of cholesterol: To make simplicity let us consider the established structure of
Side chain cholesterol followed by structural elucidation.
The established structure of Cholesterol is
22
shown in the figure:
21
23
The complete constitution of cholesterol
Angular methyl
groups 18
20 26 may be dealt in the following headings:
CH3 (A) Structure of the nucleus (B) Position of the
12 17 24 25
11 13
D
16 27 hydroxyl group and double bond (C) Nature
19 C
CH3 14 15 and position of the side chain (D) Position of
1 9 the angular methyl groups
2 10 8
A B
3 5 7 (A) Structure of the nucleus:
4 6
OH Nucleus
1.The molecular formula of cholesterol is
C27H46O.
Structure of Cholesterol
2. On acetylation it forms monoacetylate
indicating the presence of one hydroxyl group.
3. It adds up two bromine atoms suggesting the presence of one double bond.
4. Cholesterol on reduction gives cholestanol, which on oxidation with chromic acid yields
cholestanone The latter on reduction gives cholestane.
H2-Pt CrO3 Zn-Hg/HC l
Cholesterol Cholestanol Cholestanone Cholestane
C27H46O (I) C27H48O (II) C27H46O (III) C27H48 (IV)
This lead to the following conclusions:
i) The conversion of I to II proves the presence of the double bond.
ii) Oxidation of II to III (a ketone) shows that the cholesterol is a secondary alcohol.
iii) The saturated parent hydrocarbon (C27H48, IV) of cholesterol correspondings to the general
formula (CnH2n-6) for tetracyclic compounds, or the D.B.E. of cholestane is (27+1 48/2 = 28-24 = 4).
Hence, cholestane is tetracyclic; thus cholesterol is a tetracyclic alcohol.
6
 5. On selenium distillation at 360o, cholesterol gives Diel’s hydrocarbon and chrysene; the
formation of the former compound suggests that cholesterol has Diel’s hydrocarbon nucleus in its
structure.
CH3

Se
Cholesterol
360o

Diel's hydrocarbon
(3/-methyl-1,2-cyclopentenophenanthrene)

(B) Position of the hydroxyl group and double bond:

1. Cholestanone, III, on oxidation with nitric acid gives a dicarboxylic acid, V which on pyrolysis
yields a ketone, VI.
HNO3 Pyrolysis
Cholestanone Dicarboxylic acid Ketone
360o
(two isomeric products)
C27H46O (III) C27H46O4 (V) C26H44O (VI)

These observations leading to the following conclusions:

i) Since the dicarboxylic acid (V) contains the same number of atoms as the ketone (III) from
which it is derived, the keto group in (III) must therefore be in a ring [had it been in a side chain, an
acid have lesser number of carbon atom would have been formed].
ii) The conversion of dicarboxylic acid (V) to a ketone (VI) indicates that the dicarboxylic acid is
either a 1,6- or 1,7-dicarboxylic acid. According to Blance rule dicarboxylic acids upto
1,5-give anhydride on heating or pyrolysis, but 1,6- or 1,7-dicarboxylic acid yields a ketone with the
loss of one carbon atom, on similar treatment. This dicarboxylic acid is obtained from the hydroxyl
group of cholesterol which can’t be present in ring D as it would form a 1,5-dicarboxylic acid instead
of 1,6- or 1,7-dicarboxylic acid on the above treatment. Hence the hydroxyl group may be either in
ring A, B or C.
iii) The formation of two isomeric dicarboxylic acids, (V), suggest that the keto group in
cholestanone is flanked by a
methylene group on either side HNO3 HNO3
( CH2 .CO .CH2 ). HOOC HOOC
A HOOC
Examination of the proposed HOOC
O
structure of cholesterol or the
structure of Diel’s hydrocarbon Cholestanone (III)
indicates that such an
arrangement is possible only if Formation of two isomeric acids by CH2.CO.CH2
the hydroxyl group is present in
the ring A and in position 2 or 3.

7
 2. Cholestenone, III, on treatment with methyl magnesium iodide followed by selenium
dehydrogenation yields 3/,7-dimethyl-cyclopentenophenanthrene, VII, the structure of which is
proved by synthesis.
CH3
/
3

CH3MgI A Se
A H2C

O HO 7

H3C
Cholestanone (III) 3/,7-dimethyl-cyclopentenophenanthrene VII

The formation of (VII) suggests that the hydroxyl group in cholesterol is present in position 3,
which corresponds with the position 7 of (VII).
3. Position of the double bond: Consider the following set of reactions:

These reactions lead to the following conclusion:

* The conversion of I to VIII represents the hydroxylation of the double bond.
* Cholestanetriol VIII, on oxidation gives a diketone, IX, indicating that in VIII two of the
hydroxyl groups are secondary in nature and the third (resistant to oxidation) is tertiary one.
* Dehydration of hydroxycholestanedione (IX) by heating in vacue and subsequent reduction of
the double bond forms cholestanedione (X) and this on oxidation, gives a tetracarboxylic acid (XI)
without loss of any carbon atom suggests that the two ketonic groups in (X) are present in two
different rings [had they been in the same ring, then carbon atom would have been lost during
oxidation], ie., the double bond and the hydroxyl group in cholesterol are present in two different
rings and it has already seen that the hydroxyl group of cholesterol is present in ring A, therefore, the
double bond must be present either in ring B, C or D.
* Since cholestanedione, X, can form a pyridazine derivative with hydrazine, the two ketonic
groups of X are in -positions with respect to each other which is possible only if the double bond is
present in between C5 and C6 and thus all the above reactions can be written as below:

8
 H2O2 CrO3 A
A B A B B
CH3COOH
HO O
HO HO HO
I OH O
VIII IX

-H2O heat
CrO3
HOOC
A B
HOOC COOH

H COOH

XI Zn-CH3COOH A B
A B

O
O  H

O
O

NH2NH2 X
A B


N  
H

4. The position of the double bond is further proved by the following set of reactions:

Copper oxide KMnO4

Cholesterol Cholestenone Keto-acid + CO2
290o [O]
I XII XIII

Which lead to the following conclusions:

* Oxidation of XII to XIII with the loss of one carbon atom indicates that the keto group and the
double bond in XII are in the same ring. Moreover, ultraviolet absorption spectrum of XII, max 240
nm, shows that the keto group and the double bond are conjugated. These results can be explained on
the assumption that there is a migration of the double bond of cholesterol during the formation of
cholestenone, XII, which is possible only if the double bond is present in between C5 and C6.

Copper oxide KMnO4

+ CO2
290o [O] HOOC
HO O O
I XII XIII

9
 (C) Nature and position of the side-chain:
1. Acetylation of cholesterol produce cholesteryl acetate and this, on oxidation with chromium
trioxide forms a steam-volatile ketone ie. isohexylmethyl ketone and the acetate of a
hydroxyketone(which is not steam volatile).
CH3
H3C CH2-CH2-CH2-CH
Ac2O Cro3 Acetate of C CH3
Cholesterol Cholesteryl acetate a hydroxy +
ketone O

isohexylmethyl ketone

The results show that the isohexylmethyl ketone forms the side-chain of cholesterol and is attached to
the nucleus of cholesterol through the carbon atom oxidised to  C=Ogroup. These results do not
show where the side-chain is attached to the nucleus of cholesterol, but if we accept that the position
is at 17, then the reactions may formulate as follows:

2. The nature of the side-chain has also been shown by the application of the Barbier-Wieland
degradation (B-W) in the following way. Since this method also leads to evidence that shows which
ring of the nucleus is attached to the side chain. The Barbier-Wieland degradation offers a means of ‘
stepping down’ an acid one-carbon atom at a time as follows:
CH3OH 2C6H5MgBr -H2O
RCH2COOH RCH2COOCH3 RCH2C(OH)(C6H5)2 RCH=C(C6H5)2
HCl
CrO3

RCOOH + (C6H5)2CO

10
 Here cholesterol is converted into 5-cholestane, also knownas coprostane (a stereoisomer of the
cholestane, IV) and now if the whole of the nucleus of coprostane by R and the side chain by C n then
the coprostane molecule will be represented by R.Cn. Now, degrade the side-chain of this molecule
by oxidation and B-W degradation as follows:

These degradation reactions lead to the following conclusion:

* The formation of acetone from coprostane indicates that the CH3 CH3
CrO3
side chain (Cn) of the coprostane terminates in an isopropyl -CH O=C
group. CH3 CH3

* The conversion of bisnor-5-cholanic acid to the ketone,

etiocholylmethyl ketone indicates that the -carbon atom of the acid is secondary in nature, ie., there
is an alkyl group on the -carbon atom in bisnor-5-cholanic acid.

* Oxidation of etiocholylmethyl ketone to 5--etianic acid with the loss of one carbon atom
suggests that the alkyl group in etiocholylmethyl ketone and in bisnor-5-cholanic acid is methyl.
* The oxidation of etiocholanone to etiobilianic acid without any loss of carbon atom suggests that in
the former the ketonic group is present in the ring.
From the foregoing evidence it can 6 CH3 4 3 2 1
CH3
be concluded that the coprostane (and 5

hence cholesterol) contains a side R CH CH2 CH2 CH2 CH

CH3
chain of eight carbon atoms arranged
in the following order: Cn

3. Point of attachment of the side-chain to the nucleus (R):

a) The dicarboxylic acid, etiobilianic acid (obtained above), on heating with acetic anhydride
forms an anhydride suggesting that it is a 1,5-dicarboxylic acid (Blance rule). 1,5-dicarboxylic acid
can only be obtained from a five membered ring; hence the side-chain is attached to the five
membered ring D.
b) Cholesterol on selenium dehydrogenation yields Diel’s hydrocarbon indicating that the side
chain is attached to C17 in cholesterol. This position is further proved by X-ray photographic and
surface film measurements

11
 4. Thus the nature and position of the side chain is known and hence it can formulate the
conversion of coprostane into etiobilianic acid as follows [only ring D is shown in the following
figure):

COOH
CrO3 B-W
CH3COCH3 + COOH

5-Cholestane
5-Cholanic acid Nor-5-Cholanic acid
Coprostane
R.Cn-3 R.Cn-4
R.Cn

B-W

O
O COOH
COOH
B-W CrO3 B-W

Etiocholanone 5-Etianic acid Etiocholylmethyl ketone Bisnor-5-Cholanic acid

R.Cn-8 R.Cn-7 R.Cn-6 R.Cn-5

HNO3

COOH
COOH

Etiobilianic acid
R.Cn-8

(D) Position of the angular methyl groups:

The cyclopentenophenanthrene nucleus and the side-chain of the cholesterol account for 17 and 8
carbon atoms, respectively; thus 25 out of 27 carbon atoms of cholesterol have been accounted for,
the rest two-carbon atoms are found to be present as angular methyl groups by the following facts.
i) The keto-acid, XIII (obtained in B-4), on Clemmensen reduction followed by twice B-W
degradatioins gives a tertiary acid, so one of the angular methyl groups must be present on
C10.

9
10 8 Copper oxide KMnO4 9
10 8
A 5 B7 A 10
B A B + CO2
HO 290o [O] HOOC 5 7
6 O O 6
I XII XIII
Cholesterol Cholestenone Keto-acid

Zn-Hg
HCl

HOOC 9 2 B-W 9
10 8 A 10 8
B
5 B 7 HOOC 5 7

Tertiary acid

12
 ii) On selenium dehydrogenation, cholesterol yields Diel’s hydrocarbon and chrysene; the
formation of the latter is explained by the fact there is an angular methyl group at either C13 or
C14, which enters the five membered ring (D) of cholesterol to form a six membered ring of
chrysene.
22
21
23
20 26
18
CH3
12 17 24 25 CH3
11 13 16 27
19 C D
CH3
9
14 15 Se +
1
2 10 8 Dehydrogenation
A B
3 5 7
4 6
OH
Cholesterol Diel's hydrocarbon Chrysene

iii) Etiobilianic anhydride on distillation with selenium gives 1,2-dimethylphenanthrene

indicating that the methyl group is present in position 13; had it been on C 14 then only
monomethy phenanthrene would
Side chain
O
18
18
H3C H3C CH3
C O
(A) COOH 12
12
19 2219 11 13 C=O Se
11 13 COOH H3C C 14
H3C C 14 21
1 9 1 9 CH3
2 10 8
2
A
10
B
8
Angular methyl 3
A 5
B 23
3 5
6 7
20 4 6 7 26
4 groups 18 1,2-dimethyl phenanthrene
Etiobilianic acid CH3 Etiobilianic anhydride

12 17 24 25 O
11 13 16 27
(B) 19 COOH C
D C O
12
12
CH 19 13 C=O Se
H3C
19 11
C 14
13 3 COOH 14 15
H3C
11
C 14
1 19 9 2
1
10
9
8
18 CH3
18
2
A2
10
B
8
H310
C 8 3
A 5
B H3C
3 5 7
4 A6 7
B 4 6

5 Etiobilianic anhydride Monomethylphenanthrene

Etiobilianic
3 acid 7
4 6
OH Nucleus have been formed:
Stereochemistry of the steroids:
As the structural formula of the
Structure of saturated cholesterol saturated sterol shows that it has nine
dissimilar asymmetric carbon atoms (eight in the nucleus and one in side chain eg., 3, 5, 8, 9, 10, 13,
14, 17 and 20). So, there are 512 optical isomers possible according to the formula 2 n (29 = 512).

13
 Moreover, the fusion of the ring A to B, B/C and C/D may be either in cis- or trans-manner
but as the steroid molecule is essentially flat so the ring B and C are always fused in trans-
manner. It has been found that all the naturally occurring steroids except those of heart poison
belong either to the cholestane series or to the coprostane series, which have the following
types of configuration.

R R
H
H H3C
H3C
19 D
D H3C C
C
H3C H
H
H
H
H H A B
A B
H
H Coprostane
Cholestane
A/B trans A/B cis
B/C trans B/C trans
C/D trans C/D trans

Where R represents the side chain and a full line represents the group above the plane of the
molecule, and a dotted represents group below the plane. Both the angular methyl groups are
found to be cis. The compounds derived from cholestane are known as allo compounds where
as the compounds derived from coprostane are known as normal compounds. Similarly, taking
–OH group at position 3, can examplify the configuration of the substituent group. If it is
present above the plane of the ring, ie., in cis-position with respect to methyl group at C10, then
it belongs to -series. On the other hand, if hydroxyl groups are present below the plane of the
ring, it is known as - or epi-compound.

Cardiac-Active Principles: There are some steroidal compounds which when injected exert a
specific powerful action on cardiac muscles of the animals including man. These compounds are
known as cardiac-active or cardiotonic principles. Only a very small amount causes a beneficial
effect on a diseased heart but an excessive dose causes death due to hearth contraction. These steroids
are mainly found in plants as glycosides and, therefore, they are known as cardiac glycosides or
cardiac poisons. On hydrolysis they split off sugar moiety and a steroid aglycone or genin.
The important sources of these glycosides are the seeds, leaves, stems, roots or barks of plants
belonging to the families apocynaceae, lilliaceae, moraceae, rannuculaceae, etc. Other sources are sea
onion and digitalis plants. The physiological activity of the glycosides is due to aglycone only; the
sugar part renders favorable solubility and distribution characteristics to the glycosides.
Aglycons may be of two types, viz. cardenolides and bufadienoldes or scilladienolides. .
14
Cardenolides is the more common type and contain an , -unsaturated -lactone ring.They show a
max at 220 nm. On the other hand, scilladienolides is less common type and contain a -ring having a
conjugated diene system. They show a  max at 300 nm. Both the types of aglycons have the normal
configuration at C8, C9, C10, C13 and C17; and both contain a hydroxyl group at C3 and C14.. However,
the two types may differ in the configuration at C3 and C5, and in unsaturation and oxygen functions.
The configuration of C14 hydroxyl group is  in both the types. In glycosides, the C3 of aglycon is
always linked to the sugar. However, the glycosidic linkage is  for D-suigar, and  for L-sugar.
Two most common examples of cardiotonic glycosides are digitoxigenin and scillaren A.
Other examples of aglycon are strophanthidin, which is obtained by the hydrolysis of glycisides
present in the seeds of strophanthus kombe one of which is k-strophanthoside.
O
O
O O

H OH H OH
HO HO
Digitoxigenin Scillaren A

O
O

O
HC
Enzymes
HCl
H OH
-glucose -glucose cymarose O
OH
k-strophanthoside

CHO O
O
CH2

Glucose + Glucose + HCOCH3 + O

HCOH HC
HCOH H OH
CH3 HO
OH

15
cymarose strophanthoside
Hormones: Hormones are the organic substances, which are secreted by the ductless glands
(endocrine glands). These are brought to the different organs of the body by means of blood stream,
to produce the various physiological functions. These are required in small quantities and are specific
in their functions. The deficiency of a hormones leads to a particular disease (pathological condition),
which can only be cured by the administration of that hormones.

Classification: Up to date about 60

Hormones
hormones have been recognized so far,
and out of these more than thirty are
steroids, whereas the rest of all are Steroids Non-steroids
known as non-steroid hormones. The
steroids hormones are again divided into
two types: a) Sex hormones and b)
Adernal cortical hormones. Sex Sex-hormones Adernal cortical hormones
hormones are further divided into two
classes: (i) Male and (ii) Female sex
hormones. Female sex hormones are Male sex-hormones Female sex-hormones
or Androgens
again subdivided into two types
Oestrogens and Gestogens, respectively. Oestrogens Gestrogens

Sex-hormones: These are produced in the gonads (testes in male and ovaries in female). Their
production is stimulated by another group of hormones secreted by the anterior lobe of the pituitary
gland and carried to the gonads through the blood stream. This has been proved experimentally by
removing the anterior lobe of the gland in an animal, which fails to show sexual activity, but by
implantation of the gland, the animal restores the normal sexual characteristics. Due to this fact the
sex-hormones are sometimes called the secondary sex-hormones and sex-hormones of the anterior
lobe of the pituitary glands are called the primary sex-hormones or gonadotropic hormones.
Male sex-hormones or androgens consist of testosterone and androsterone while one of the female
sex-hormone oestrogens consist of oestradiol, oestrone, oestriol etc., while progesterone is the
important member of the other female sex-hormone gestogens. The testes of male secrete testosterone
while the ovary of female produces mainly oestradiol and progesterone. These three hormones are the
most important and rest of all are the metabolic products of these three hormones, eg., oestrone,
oestriol and other oestrogens are the metabolites of oestradiol and androsterone is metabolite of
testosterone.
The sex-hormones are responsible for sexual process, and for the secondary characteristics which
differentiate males from females, eg., texture and distribution of hair, texture of the skin, character of
voice, etc. Synthetic substitutes for all the three hormones have been prepared, eg, ethinyl oestradiol,
ethisterone, and methyltestosterone for oeatradiol, progesterone and testosterone, respectively.
Adrenal cortical hormones: There is a small gland over the kidney known as adrenal gland.
It is divided mainly into two parts: the cortex (outer) and medulla (inner). The latter secretes mainly a
hormone known as adrenaline, whereas the cortex secretes mainly cortical hormones.
The corticoids have many physiological functions, but their main functions are the control of
carbohydrate and protein metabolism and the control of the balance of water and electrolytes.
They may be classified into four groups according to their structure and biological function.
(i) Oestrogens, (ii) Androgens, (iii) Progesterones and (iv) Corticoids. Out of these sex organs mainly
secrete the first three types so they belongs to sex hormones whereas the corticoids are the
characteristic adrenal cortical hormones. The production of all of these hormones is controlled by the

16
hormones produced by the anterior lobe of pituitary body and are known as adrenocorticotropic
hormones (ACTH).
Isolation: The adrenal gland is extracted with alcohol or acetone. From the solution so obtained
adrenaline is removed by taking the advantage of its basic properties. Now the mixture of adrenal
cortical hormones (after removing sex hormones) are separatyed from each other by means of
combination of methods viz., fractional crystallization, partition coefficient and application of
Grignard’s T or P reagent. Further purification is done by the chromatographic methods
(adsorption or partition chromatography). As these hormones are very sensitive to acids as well as
alkalis so strong acidic or alkaline conditions should be avoided during their isolation.
By the application of these methods, forty steroids have been isolated from the cortex of adrenal
body. Out of these forty, only eight are found to be biologically active. The eight biologically active
hormones are mentioned in the figure.

CH2OH CH2OH CH2OH

CO CO CO
18 18 18 OH
HO 12 17 O 12 17 HO 12
17
16 16 16
13 13 13
11 11 11
1 19 1 19 1 19
14 14 14
15 15 15
2 9 2 9 2 9
10 8 10 8 10 8

3 5 7 3 5 7 3 5 7

O 4 6 O 4 6 O 4 6

Corticosterone; 11-dehydrocorticosterone; 17-hydroxy corticosterone;

11, 21-dihydroxyprogesterone; 21-hydroxy-11-keto-progesterone; cortisol;
substance H compound A substance M

CH2OH CH2OH CH2OH

CO CO CO
18 OH 18 18 OH
O 12 12 HO 12
17 17 17
16 16 16
13 13 13
11 11 11
1 19 1 19 1 19
14 14 14
15 15 15
2 9 2 9 9
10 8 10 8 2 10 8
3 5 7 3 5 7 3 5 7
O 4 6 O 4 6 HO 4 6

11-dehydro-17-hydroxy corticosterone; 11-deoxycorticosterone; substance C

cortisone; 21-hydroxyprogesterone;
substance F or compound E substance Q
CH2OH CH2OH
CO HO
18
CO
18 OH
12 O HC
17 17
16 12 16
13 13
11 11
1 19 19
14 1 14
15 15
2 9 9
10 8 2 8
10
3 5 7 3 5 7
O 4 6
O 4 6

11-deoxy-17-hydroxy corticosterone; Aldosterone

cortexolone; 17
substance S
 Relation of structure to physiological activity: For all the naturally occurring adrenal cortical
hormones two structural features are essential for their biological actions. (a) Presence of an
, -unsaturated ketonic group where the double bond between C4 and C5 and a ketonic group at C3
and (b) presence of a ketonic group at C20. However, the other functional groups on other positions
also exhibit some specific and important functions, eg, (i) hydroxyl group at C 21 increases sodium
retention and is necessary for carbohydrate metabolism, ii) presence of either –OH or –CO at C11
is necessary for carbohydrate activity and decrease sodium retention. iii) hydroxyl group at C 11
increases carbohydrate activity. On the basis of the point of consideration of (ii) the seven
biologically active hormones can be divided into three main types: (a) which are oxygenated at C11
eg. The first four corticoids those help in carbohydrate and protein metabolism. (b) which are not
oxygenated at C11, eg, 11-deoxy-17-hydroxycorticosterone. These hormones are not active in
carbohydrate and protein metabolism. (c) which is oxygenated at C11 and its methyl group at C18 is
replaced by an aldehyde group, eg, aldosterone. This hormone affects both mineral (Na, K, Cl, etc.),
and organic metabolism.
So the main functions of the corticoids are to affect the carbohydrate, lipids, protein and the
mineral, eg, Na, K, Cl etc., metabolism. In the first case cortisol is the most active and in the latter
case aldosterone is the most effective. So the two may also be known as major glucocorticoid and the
major mineralocorticoid, respectively. The deficiency of corticoids also leads to muscle weakness,
decrease in resistance to shock, eg., cold, mechanical and chemical. The decrease in carbohydrate
metabolism leads to Addison’s disease.
Properties: All are colorless compounds, dextrorotatory and are very sensitive to alkali.
They readily reduce alkaline AgNO3 solution due to the presence of -ketol (-hydroxy ketone)
group (–CO.CH2OH). The hydroxyl group at C21 behaves in the usual way. The ketonic group at C11
does not give normal ketonic reactions because of steric effects, eg., it does not form oxime,
phenylhydrazone, semicarbazone, etc. The 11-keto group is resistant to catalytic reduction in neutral
solution, but can be reduced in acid solution; it is readily reduced by LiAlH4 to a hydroxyl group; and
to methylene group by Clemmensen reduction.

18