ISOLATION, I DENTI FI CATI ON, AND CHEMISTRY OF THE
ANTIBACTERIAL ALKALOID SOLANOCAPSINE
FROM SOLANUM PSEUDOCAPSICUM L.

by
P e r Msz^lgaard B o l l

A THESI S
S u b m itte d t o the School of Advanced G ra duate S t u d i e s of Michig an
S t a t e U n i v e r s i t y o f A g r i c u l t u r e and A p p l i e d S c i e n c e
in p a r t i a l f u l f i l l m e n t of t h e r e q u i r e m e n t s
fo r the degree of
DOCTOR OF PHILOSOPHY

Department

of C h e m i s t r y

1957

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ACKNOWLEDGEMENTS
The a u t h o r w i s h e s
to Dr.

H a n s A.

patience

to e x p re ss

Lillevik,

whose

t h i s work,

of Drug P l a n t R e s e a r c h ,

greatly

and t o D r .
Department

initiation

of

t h e p r o b l e m a nd f o r

the

of

the

course

Appreciation
me n t
R.

cooperation

also

of

use of v a r i o u s

advice

the

for

with his

facilitated

E. H.

Lucas,

helpful

advice

the

Professor

of H o r t i c u l t u r e ,

to

t h e member s o f

from time

for

during

to

Department

time

the D epart­
and t o Dr .

of H e a l t h f o r

the b a c t e r i o l o g i c a l

author wishes

to

t h a n k Dr .

of A g r i c u l t u r a l

testing.

H. M.

Sell,

Chemistry,

equipment.

F i n a l l y , - he w i s h e s
Health

together

the Michigan Department

Research Professor,
for

i s due

in c o n n e c t i o n w i t h

In a d d i t i o n ,

appreciation

study.

of Chemistry fo r

Y. G o t t s h a l l

sincere

interest

and e n c o u r a g e m e n t have

com pletion of

his

providing

to

funds

thank the National
in support

Institute

of t h i s work.

of

VI TA
Per M^Igaard Boll
candidate

for

Doctor

Final

examination:

Dissertation:

Outline

the

degree

of

of P h i l o s o p h y

F e b r u a r y 25, 1957, 10:00 A. M. , C o n f e r e n c e
Room, K e d z i e C h e m i c a l L a b o r a t o r y .

I s o l a t i o n , I d e n t i f i c a t i o n , a n d C h e m i s tr y of
A n t i b a c t e r i a 1 A l k a l o i d S o l a n o c a p s i n e from
S o l a n u m p s e u d o c a p s i cum L .

the

of S tu d ies:

Major

subject:

Minor

subjects:

Biographical
Born,

Organic Chemistry,

1 9,

Undergraduate

1929,

Herning,

Studies:

Studies:

Experience:

Chemistry

D e n ma r k

E x am in a tu sp h a rm a c iae , Royal Danish
S c h o o l o f P h a r m a c y , 19Jf8 - 1950 .

C a n d id a tu s pha rm a ciae, Royal Danish
School of Pharmacy, 1950-1952;
M i c h i g a n S t a t e U n i v e r s i t y , 1951+- 1957.

Pharmaceutical Chemist, F re d e rik sb e rg
H o s p i t a l , Co pe n hage n, Denmark, 1 9 5 2 - 19 5 3 ;
P h a r m a c e u t i c a l C h e m i s t , Leo P h a r m a c e u t i c a l
Product^, Cope nhage n, Denmark, J a n u a r y A p r i l , 195U-; S p e c i a l G r a d u a t e R e s e a r c h
A s s i s t a n t , Michigan S tate U n i v e r s i ty , A p r il,
195^--December, 1955, S e p t e m b e r , 1 9 5 6 - F e b r u a r y , 1957; G r a d u a t e T e a c h i n g A s s i s t a n t ,
Mi chiga n S t a t e U n i v e r s i t y , J a n u a r y , 1956J u n e , 1956.

Member o f A m e r i c a n C h e m i c a l
Sigma X i ,

Analytical

Items:

July

Graduate

Biochemistry

Society,

and Danmarks f a r m a c e u t i s k e
iii

S o c ie ty of

Selskab.

the

ISOLATION, IDENTI FI CATI ON, AND CHEMISTRY OF THE
ANTIBACTERIAL ALKALOID SOLANOCAPSINE
FROM SCLANUM PSEUDOCAPSICUM L.

by
Per M^Igaard Boll

AN ABSTRACT
S u b m i t t e d to the School of Advanced G r a d u a t e S t u d i e s of Mic h ig a n
S t a t e U n i v e r s i t y o f A g r i c u l t u r e a nd A p p l i e d S c i e n c e
in p a r t i a l f u l f i l l m e n t of t h e r e q u i r e m e n t s
f o r t h e d e g r e e of
DOCTOR OF PHILOSOPHY

Department of Che mistry
Ye a r

Approved

1957

P e r Mj/lgaard B o l l
I t was o b s e r v e d
ous e x t r a c t

of

antibacterial
isolate

the p l a n t

and c h e m i c a l l y

associated

identified

as

(2).

wa s 3

for mice

injected

for

for

both these

this

possessed

s u b s t a n c e ( s ).

fraction

alkaloid

culture

of

the p la n t

so 1 a n c a p s i n e .

broth.

The m e t h o d

used by S c h l i t t l e r

activity

of pure

a n d 200 mg.

lethal

for

its

hydrochloride.

d o s e wa s 300 m g . / k g .

to

soiancapsine

of body w e i g h t

a colored p rin cip le

of

this

from the

colored

alumina,

acid

s ho we d t h a t

the

cent

(based

2 . 1 5 Pe r

cent

in l e a v e s p l u s

So 1 a n o c a p s i n e

Solanum whi ch h a s n o t

content

on d r y w e i g h t )

is the
been

isolate

and p u r i f y

a nd Dowex 5 0 .

on p l a n t s

alkaloid

2 . 2l+ p e r

No f u r t h e r

i m p u r i t i e s by column c h r o m a t o ­

alumina,

determinations

inhibiting

s u b s t a n c e was u n d e r t a k e n .

A t t e m p t s w i t h o u t much s u c c e s s w e r e made t o

period

dose

compounds.

wor k on t h e p u r i f i c a t i o n

Alkaloid

solan-

The mi n i mu m l e t h a l

g r o w t h o f D i p l o c o c c u s p n e u m o n i a e wa s f o u n d p r e s e n t .

graphy using

and

p e r i t o n e a l l y wa s 5 0 - 1 0 0 m g . / k g . o f b o d y

the

In a d d i t i o n

so 1a n o c a p s i n e

an a q u e ­

s t u d y was t o

antibacterial

The a n t i t u b e r c u 1 a r

so 1a n o c a p s i n e

Subcutaneously,

of

i s o l a t i o n wa s t h a t

if/m l,

ocapsine

weight

for

the

alkaloid

steroidal

with m odifications

that

a g a i n s t M y c o b a c t e r Lum t u b e r c u l o s i s wa s

w ith the

the

and U e h l i n g e r

The p u r p o s e

identify

active

(1)

S o l a n u m p se u d o c a p s i cum L.

properties.

A substance
found

by L u c a s and a s s o c i a t e s

g r o wn o v e r
varied

in r o o t s

a three

from 0. 83

year
to

and f r o m 1 . 5 3 t o

stems.

only alkaloid
isolated

from the

plant

as a g l y c o s id e .

genus

Therefore,

2
the p o s s i b i l i t y of
being

acted

fully

examined.

in a b s o l u t e
in th e

an e x t r a o r d i n a r y l a b i l e

upon e n z y m a t i c a l l y or
Enzyme

ethanol

or

in b o i l i n g w a t e r

that

did

so 1a n o c a p s i n e

the p l a n t s

and u s i n g weak a c i d s

not y ie ld

is p r e s e n t

bond

i s o l a t i o n was c a r e ­

i n a c t i v a t i o n by p l a c i n g

iso la tio n procedure

regarded

during

glycosidic

a glycoside.

in t h e p l a n t

It

is

as a f r e e

alkaloid.
It

has been re p o rte d

ocapsine
the

contains

alkaloidal

Paper
spot

glycoside

corresponding

found

to the

be a c r u d e

crude

amorphous
to

Of t h e s e ,

solanidine

activity.

only

Tomatine

and

in a d d i t i o n

solan-

so 1 a n o c a p s i d i n e

its

solvents

aglycone

and F r a e n k e 1- C o n r a t

(3),

solanidine.

revealed

so 1 a n o c a p s i n e .

alkaloid

to

o n l y one
Solanocap(3)

preparation,

wa s

which

so 1a n o c a p s i n e .

alkaloid preparation

lated.

found

and

alkaloid

by B a r g e r

c o u l d be c r y s t a l l i z e d
Solanine

solanine

in d i f f e r e n t

as d e s c r i b e d

to

the p la n t

an a mo r p h o u s a l k a l o i d

chromatography

sidine,

that

f r o m Sol anum t u b e r o s u m and a

from Solanum c a r o l i n e n s e

solanine

were

iso­

s howe d we a k a n t i t u b e r c u l a r

f r o m L y c o p e r s i c o n e s c u l e n t u m was

likewise

inactive.
Some c h e m i c a l

undertaken.

structural

s t u d i e s on so 1 a n o c a p s i n e wa s

Two d e r i v a t i v e s p r e v i o u s l y

N,N'-diacetylsolanocapsine

0 , N - d i a c e t y l s o 1a n o c a p s i n e
The p i p e r i d i n e
hydrogenolysis.

ring of

the

thought both

wa s f o u n d mo r e

t o be

identifiable

as

and 0 , N , N t - t r i a c e t y l s o l a n o c a p s i n e .
s o l a s o d a n n u c l e u s wa s o p e n e d by

3

Schlittler
ture

for

groups

and U e h l i n g e r

so 1a n o c a p s i n e

unassigned.

concluded

t o be

visionally
proposed

for

left

proposed

a solasodan

3 position

and t h e

to the 8 p o s i t i o n .

sol ano ca p s ine

struc­

t h e p r i m a r y a mi n o a n d h y d r o x y l

Fr o m t h e p r e s e n t w o r k t h e

in the

assigned

and

(2)

a mi n o g r o u p w a s

hydroxyl

group p r o ­

Th e t e n t a t i v e

structure

i s 3o<_-am i n o so 1 a s o d a n - 8 - o 1 m o n o -

hydrate .

1. F r i s b e y , A . , R o b e r t s , J . M. , J e n n i n g s , J . C . , G o t t s h a l l ,
R. Y . , a n d L u c a s , E . H „, Q u a r t . B u l l . M i c h . A g r . E x p . S t a . ,
M i c h . S t a t e C o l l . , V o l . 3 5 , 3_, 3 9 2 —ipOIp ( 1 9 5 3 ) .
35,

2. S c h l i t t l e r , E . ,
2031^-20104- ( 1 9 5 2 ) .

3. B a r q e r , G . ,
1936, 1 5 3 7- 15 02 .

a nd U e h l i n g e r ,

H.,

Helv.

a nd F r a e n k e 1 - C o n r a t , H. L . ,

C h i m. A c t a ,
J.

Chem.

Soc.

TABLE OF CONTENTS
Page
I.

INTRODUCT I O N .....................................................................................................

I

II.

H I S T O R I C A L ..........................................................................................................

2

A. H i s t o r y o f A n t i b a c t e r i a l A g e n t s f r o m H i g h e r
P 1a n t s
.........................................................................................................
..................................
1. D e f i n i t i o n o f A n t i b i o t i c A g e n t s
2 . E a r l y D e v e l o p m e n t s ....................................................................
3 . Mo d e r n D e v e l o p m e n t s
...............................................................
1+. P l a n t s P o s s e s s i n g A n t i b a c t e r i a l A c t i v i t y . . .

2
2
3
5
7

B.

C h e m i s t r y and P h a r m a c o l o g y of So l a n u m
p s e u d o c a p s i c u m L ....................................................................................

10

Chemical

on So I a n o c a p s i n e .....................................

12

EXPER I ME N T A L ....................................................................................................

16

A.

Apparatus

.

16

B.

Reagents,

Materials,

C.

Experimental

C.
III.

Studies

a nd A n a l y t i c a l

Procedures

Methods

. . .

18

..............................................................

25

ISOLATION W O R K .......................................................................
1.

2.

25

P relim in ary Experiments
.....................................................
a . E x t r a c t i o n a g e n t s ...............................................................
b. P o s s i b i l i t y o f an a n t i b a c t e r i a l a l k a l o i d
,
c. F r a c t i o n a t i o n using ion-exchange
.....................
.........................
d. E x t r a c t i o n o f b a s i c p r i n c i p l e s
I s o l a t i o n a n d P u r i f i c a t i o n o f So 1a n o c a p s i n e
.
a. A n t i b a c t e r i a l a c t i v i t y in e x t r a c t s of
r o o t s a n d l e a v e s .................................................................
b.
I s o l a t i o n of crude
a l k a l o i d from r o o t s
. .
c.
I s o l a t i o n o f c r u d e so 1a n o c a p s i n e
from
leaves
.....................................................................
d . The c o m p l e t e n e s s o f a l k a l o i d e x t r a c t i o n . .
e . P u r i f i c a t i o n o f so 1 a n o c a p s i n e ...................
38
f . Co l u mn c h r o m a t o g r a p h y on a l u m i n a , Dowex
50 and ac id a l u m i n a
..........................
g . A l k a l o i d c o n t e n t i n t h e p l a n t ................................
h . D e r i v a t i v e s o f r o o t and l e a f a l k a l o i d . . .
iv

25
25
26
27
30
31
31
32
35
37
1+0
1+9
52

V

Page
3.
1|_.
5.

A n t i b a c t e r i a l S p e c t r u m o f So 1a n o c a p s i n e . . .
To x i c i t y , S t ud i e s on So 1 a n o c a p s i n e .........................
A t t e m p t s t o I s o l a t e an A l k a l o i d a l G l y c o s i d e .
a . P o s s i b i l i t y o f a g l y c o s i d e .....................................
b . P a p e r c h r o m a t o g r a p h y .....................................................
c . I s o l a t i o n a t t e m p t s ..........................................................
6. P r i n c i p l e w i t h A n t i b a c t e r i a i A c t i v i t y a g a i n s t
D. p n e umon i a e ..............................................................................
7. Ant i t u b e r c u l a r A c t i v i t y of O t h e r Solanum
A l k a l o i d s .............................................................. ] [ ] . . .
a . To ma t i n e ....................................................................................
b . S o l a n i n e a nd s o l a n i d i n e ...............................
c . C r u d e a l k a l o i d p r e p a r a t i o n f r o m S_.
carolinense
. . . . . .

CHEMICAL STUDIES ON SOLANOCAPSINE
8.
9.

10.
11.

12 .

13.
IV.
V.

................................

P h y s i c a l C o n s t a n t s ....................................
a . So 1 a n o c ap s i n e ...................................................................
b . So 1a n o c a p s i n e d i h y d r o c h l o r i d e .........................
R e a c t i o n w i t h N i t r o u s A c i d ..........................................
a . N i t r o s o so 1a n o c a p s i n e ...................................................
b . D i g i t o n i d e o f n i t r o s o so 1a n o c a p s i n e . . . .
c . D i h y d r o so 1 a n o c ap s i n e ...................................................
d . ffN i t r o s od i h y d r o so 1 a n o c a p s i n e t f ..........................
A c e t y l a t i o n S t u d i e s .....................................................................
a . 0 ?N - d i a c e t y l s o 1a n o c a p s i n e ..........................................
b . 0 , N ?Nf - t r i a c e t y l s o 1a n o c a p s i n e ..........................
M i s c e l l a n e o u s D e r i v a t i v e s ...............................................
a . Tr ime t h y l so 1 a n o c ap s i n e ..............................................
b . So 1a n o c a p s i n e p i c r a t e ..............................................
c . So 1a n o c a p s i n e o x a l a t e
. . . .
d . I s o p r o p y l i d e n e s o 1a n o c a p s i n e ...............................
e . A t t e m p t e d t o s y l a t i o n ...................................................
f . Attempted dehydration
..............................................
O x i d a t i o n S t u d i e s ...............................
a . O p p e n a u e r o x i d a t i o n o f n i t r o s o so 1a n o ­
cap s i n e
...............................................................
b. Chromic a ci d o x i d a t i o n of t r i a c e t y s o l a n o capsine
...............................................................
C o l o r R e a c t i o n s ..........................................................................

DISCUSSION AND CONCLUSIONS

..........................................................

53
5 k55
56
56
59
62
65
65
65
67
69
69
69
73
75
75
78
78
79
8l
8l
85
87
87
88
88
88
91
92
92
93
93
98
99

SUMMARY........................................................................................

B I B L I O G R A P H Y ..............................................................................................................
APPENDIX I

..............................................................................................................

APPENDIX I I ...................................................................................................................

120
129
131

L I S T OF TABLES
Page
R e s u l t s of V a r io u s E x t r a c t i o n S o l v e n ts w ith R o o ts
o f S . p s e u d o c a p s i c u m ..........................................................................

26

A n t i b a c t e r i a l A c t i v i t y of F r a c t i o n s O b t a i n e d by
I o n - E x c h a n g e ...............................................................................................

29

A n t i b a c t e r i a l A c t i v i t y of F r a c t i o n s O bta ine d from
R o o t E t h a n o l i c E x t r a c t ....................................................................

31

A n t i b a c t e r i a l A c t i v i t y i n E x t r a c t s f r o m R o o t s a nd
L e a v e s . .........................................................................................................

32

A n t i b a c t e r i a l A c t i v i t y During Root A l k a lo i d
E x t r a c t i o n ....................................................................................................

35

A n t i b a c t e r i a l A c t i v i t y D u r i n g So 1a n o c a p s i n e
Pur i f i c a t ion
.........................................................................

ko

.

.

.

Data f o r E l u t i o n C hr om atography of E x t r a c t from
R o o t s o f S_. p s e u d o c a p s i cum on A 1 urn i n a
..........................

6-3

Data f o r E l u t i o n C hr om atography of Crude S o l a n o c a p s i n e f r o m R o o t s on A c i d A l u m i n a .....................................

1+8

A l k a l o i d C o n t e n t i n _S. p s e u d o c a p s i c u m on D r y
We i g h t B a s i s ...............................................................................................

5i

Chemical

.

52

A c t i v i t y o f C r u d e So 1a n o c a p s i n e A g a i n s t V a r i o u s
M i c r o o r g a n i s m s .........................................................................................

53

R ^ - V a i u e s f o r So 1a n o c a p s i n e , C r u d e L e a f A l k a l o i d ,
and Cr ude R o o t A l k a l o i d
...............................................................

58

Data C oncerning Attem pted I s o l a t i o n of A l k a l o i d
Glycoside
....................................................................................................

61

R ^ - V a l u e s o f C o mp o u n d s I s o l a t e d b y a S t a n d a r d
Procedure for A lkaloidal Glycosides
................................

62

A n t i b a c t e r i a l A c t i v i t y of F r a c t i o n s During I s o l a ­
t i o n o f P r i n c i p l e A c t i v e A g a i n s t D. p n e u m o n i a e . .

66

Constants

of Root

vi

and L e a f A l k a l o i d

.

.

-

vii
Table
XVI .
XVII.
XVIII.

Page
A n t i b a c t e r i a l A c t i v i t y of D i f f e r e n t Solanum
A l k a l o i d s .........................................................................................................

68

F r a c t i o n a t i o n o f D i a c e t y 1 so 1a n o c a p s i n e b y Co l u mn
C h r o m a t o g r a p h y on A l u m i n a .............................................................

82

F r a c t i o n a t i o n o f T r i a c e t y l s o 1a n o c a p s i n e
C h r o m a t o g r a p h y on A l u m i n a ........................

86

b y Co l u mn

LI S T OF FIGURES
F igure
1.
2.
3.
1|.

Page
Sc he me o f I o n - E x c h a n g e F r a c t i o n a t i o n o f R o o t
E x t r a c t ..............................................................................................................
Sc he me o f

Isolation

of A lk a lo id

from R o o t s

27

. . . .

33

Data f o r E l u t i o n Chromatography of E x t r a c t from
R o o t s o f S_. p s e u d o c a p s i c um on A l u m i n a ..............................

6.i|

C o l o r I n t e n s i t i e s a n d Am o u n t s o f D r y M a t e r i a l i n
F r a c t i o n s O b t a i n e d b y E l u t i o n C h r o m a t o g r a p h y on
Dowex 5 0 .........................................................................................................

6-6

o f So 1a n o c a p s i n e ..............................

71

o f So 1 a n o c a p s i n e ...................................

72

5.

Infrared

Spectrum

6.

T i t r a t i o n Curve

7.

In fra re d Spectrum
o f So 1 a n o c a p s i n e D i h y d r o c h 1 o r i d e ........................................................................................................

76-

8.

Infrared

S p e c t r u m o f N i t r o s o s o l a n o c a p s i n e .....................

76

9.

U l t r a v i o l e t S p e c t r a o f N i t r o s o s o l a n o c a p s i n e and
"N i t r o s o d i h y d r o so 1 a n o c ap s i ne , f ....................................................

77

I n f r a r e d S p e c t r u m o f " N i t r o s o d i h y d r o s o 1a n o c a p s i n e f f ...................................................................................................................

80

10.
11.

Infrared

.

83

1 2.

In f ra re d Spectrum
o f 0 , N , N - T r i a c e t y 1 so 1a n o c a p 3 i n e ...................................................................................................................

81|

13.

Infrared

89

16-.

I n f r a r e d S p e c t r u m o f Tr ime t h y l so 1 a n o c ap s i ne
H y d r o c h l o r i d e ..............................................................................................

90

I n f r a r e d Spectrum of D e r i v a t i v e O b ta in e d from
O p p e n a u e r O x i d a t i o n o f N i t r o so so 1 a n o c a p s i n e . . . .

96-

U l t r a v i o l e t Spectrum of O x i d a t i o n P r o d u c t of
T r i a c e t y l s o 1a n o c a p s i n e ....................................................................

96

R e a c t i o n o f So 1a n o c a p s i n e a n d D i h y d r o s o 1a n o c a p s i n e
w i t h N i t r o u s A c i d ....................................................................................

97

15.
16 .
17.

Spectrum

Spectrum

o f 0 , N - D i a c e t y l s o 1a n o c a p s i n e

o f T r i m e t h y l s o 1a n o c a p s i n e

v i ii

.

. . . .

I.

INTRODUCTION

The d i s c o v e r y a n d d e v e l o p m e n t
typified

by p e n i c i l l i n

discoveries
treatment

and

of

b i o t i c work
objective

since

of n a t u r a l l y
This
Several
tracts

inventions

diseases

the

has e c l ip s e d
the

safe,

occurring

from larg e

the

a nd

isolation

antibacterial

of

They f ou nd t h a t
antibacterial
appeared q u ite

of

on an e x t e n s i v e
seed p l a n t s

principles

and r a p i d

Mos t

anti­

has had as

some o f

the

its

hundreds

to microorganisms.

activity
plants

s hown b y e x ­

have been p u b l i s h e d .
) have c o n ­

screening program w ith
possessing

antibacterial

a g a i n s t M y c o b a c t e r Turn t u b e r c u l o s i s .

a considerable

activity.
promising

n u mb e r

Of t h e s e ,
(32).

identifying

of p l a n t s

s h o we d

On t h e b a s i s

of

these

the purpose

the main a n t i b a c t e r i a l

in t h e p l a n t .

1

such

S o l a n u m p s e u d o c a p s i c u m L.

i n v e s t i g a t i o n wa s u n d e r t a k e n w i t h

and c h e m i c a l l y

effective

( 3 2, 3 3 , 3L|., 35, Ij-0, lj.1, 5 4 ^ 5 5 ,

finding

especially

previous

compounds.

antibacterial

numbers of h i g h e r

and r e p o r t e d

agents,

all

of p e n i c i l l i n

search has not been r e s t r i c t e d

the purpose

this

isolation

discovery

surveys of

activity,

for

antibiotic

caused by m ic ro o rg a n is m s .

the

L u c a s and a s s o c i a t e s
ducted

(30,31),

of

of

findings
isolating

principle

or

II.

HISTORICAL

A . H i s t o r y of A n t i b a c t e r i a l
1.

Agents from Higher P l a n t s

D e f i n i t i o n of A n t i b i o t i c

A ntibiotic

agents

chemotherapeutic

are,

agents.

in the

The

the

concept

Pasteur

since

in

In t h i s
a c o mp o u n d
whereas

the

hibition
It

though
use
to

inhibiting
agent

1877

is w ithout

indicate

effect

body.

antibiotic

is e f f e c t i v e

hope

use of

them.

The

suggested
the

for

the

agent

that

antagonism

is connected

this
or

point

to

is too

However,

presence
tissue

its value

that

antibacterial

in v ivo or

activity.

to

is a p p li e d

to

growth of b a c t e r i a ,

_In_ v i t r o a c t i v i t y

in the

s u ffic ie n tly non-toxic
istration,

at

a s an a n t i b i o t i c

animal

(68)

associa­

growth

in­

and o t h e r m i c r o o r g a n i s m s .

s h o u l d be e m p h a s i z e d

in the

stance

principle)

of both b a c t e r i a

it

f r o m t he word

o f one o f

term ant ib ac te r ial

term a n t i b i o t i c

can be c l a s s e d

the word,

organisms.

study the

(or

of

antagonistic

detriment

i n f e c t i o n s m i g h t be c o m b a t e d b y m a k i n g
between d i f f e r e n t

sense

is derived

as the

tio n between organisms to
i s n o t n e w,

truest

term

a n t i b i o s i s w h i c h ma y be d e f i n e d

a nd A n t i b i o t i c A g e n t s

2

agent

toxic

alone

is

even

to perm it
sufficient

if

the

sub­

of body f l u i d s

and

is

cells

only

to p e rm it

as a m ed icin a l

ta ined.

a substance

safe

admin­

c a n be e n t e r -

3

2.
R e c o g n i t i o n of

E arly Developments

the medicinal

value

occurred very early

in t h e h i s t o r y

earliest

to

in t h e

references

ancient

described
who

is

Chinese

in Pen-1sao

said

centuries

to have

i n 1597 A . D .

literature

(65)*

One o f

Plant

included

is found

d r u g s were

In t h e

i n f o r m a t i o n was c o l l e c t e d

Chinese m e d i c i n a l s which

the

w r i t t e n by Shen-nung,

3000 B. C.

Li S h i h - c h e n p u b l i s h e d

plants

use of p l a n t s

(the Great Herbal)
about

certain

of mankind.

the medicinal

lived

additional

of

in C h i n a ,

a 52-volume

several

following

series

plants

used

a nd

on
against

s e p t i c wou nd s .
Another
the

early

sixteenth

century

i n a t o mb a t T h e b e s
hundred h e rb a l
still

reference
is the

to p la n t

E b e r s P a p y r us

in 1873.

remedies,

drugs

It

from ap p ro x im ate ly
(26),

d e s c r i b e s mo r e

i n c l u d i n g ma n y t h a t

discovered
than

are

seven

familiar

a nd

used today.
In t h e O l d T e s t a m e n t mo r e

vention
Even t h e

of

the

d e ity than

instructions

He ze k ia h have

of

a s o me wh a t

on u s e o f d r u g s
Isaiah

T a k e a lump o f f i g s .

boil,

a n d he r e c o v e r e d . "

but

medical

thought evolved,

considered

in G r e e c e ,

to

date

and

for

laid

saving

character:

along with

later

i n Rome ,

the

time

the
"And

laid

inter­
the

life

sick.
of

Isaiah
it

on t h e

kingdom of Egypt

a new s c h o o l

and mo d e r n m e d i c i n e

from the

on t h e

in h e a l i n g

And t h e y t o o k a n d

decayed

itself,

(12)

arbitrary

said,

Egyptian medicine

s t r e s s was

of

is g e n e r a l l y

of H i p p o c r a t e s

(Lj.60-361 B . C . ) .

4
(307 to 2^6 B.C. )

Theophrastus
described

the

therapeutic

193 A . D . ) a d v o c a t e d

uses of p l a n t s

in h i s w r i t i n g s

place

of m ineral

preparations

Greek

surgeon to

the

the

during

the

the

and G a l e n

(3).

re ig n of T ib e r iu s ,

to

who

in

Dioscor ides,

dealt with medicinal

Celsus,

us e o f a number o f p l a n t s

(103

use of v e g e t a b l e

in m e d i c i n e .

a r my o f N e r o ,

i n De u n i v e r s a me d i c i n a
lived

i n h i s H i s t o r i a P 1 a n t a r uin

is b e l i e v e d

mentioned

in p r o v i d i n g

a
plants

t o have

i n De M e d i c i n a

remedies for

purul­

e n t wounds.
Hippocrates
cine

for

thinkers

and G a l e n t o g e t h e r

some f i f t e e n

centuries.

and e x p e r i m e n t e r s

a nd b e c a m e

distorted

dominated European medi­

In t h e

their

absence

teaching

and m i s u n d e r s t o o d .

turned

same p e r i o d

or

According

were

stamped w i t h

were

intended.

that

if

Hence,

i t was

be v a l u a b l e
this

purpose

ma n y p e o p l e
accessible

to

doctrine,

sign of

across,

the

By t h e

and

the

its

of

"four
In th e

medicinal

plants

which they

name t o

the f a c t

resembles

a seal.

t h e p l a n t would

i t was w i d e l y u s e d f o r

sixteenth

century the

firmly established.

The h e r b a l

Paracelsus

of the

signatures”

use f o r

section

that preparations

a n d ma n y c e n t u r i e s w e r e
form.

the

o we s

of

dogma

and d r y n e s s .

" d o c t r ine of

this

" s e a l i n g ” wounds,

(91).

a l i s m h a d b e c o me

the

So 1o mo n * s S e a l

is cut

supposed

for

moisture,

some o b v i o u s

Thus,

root

cold,

1^00 A. D.

until

wa s p r o p a g a t e d .

its

heat,

into

The h e a l t h

b o d y w a s t h o u g h t b y ma n y t o d e p e n d on a b a l a n c e
principles,” i.e.,

of o r i g i n a l

recorded

(11^93-15^1)

art

of h e r b ­

re m e d ie s of

in f a i r l y

in h i s

readily

extensive

5
writings,
istry

began to bridge

and t h u s f o u n d e d

1 ishman,
in

1597.

its

gap b e t w e e n m e d i c i n e

iatrochem istry.

It

is a c u r i o u s book,

a m ix t u r e of

and c o n t e m p o r a r y f o l k l o r e .

c o n t e n t may be

gathered

o f P 1a n t e s

accurate

Some i d e a o f

from the

and c hem­

John G e r a r d e , an Eng-

w r o t e The He r b a l 1 o r Ge ne r a i l H i s t o r i e

cription
of

the

following

the

des­
nature

quotation

(37) :
D i o s c o r i d e s w r i t e t h t h a t t h e y e l l o w e W a l l f l o w e r i s most
u s e d i n p h i s i c k e , a n d mo r e t h a n t h e r e s t o f s t o c k e g i l l o f l o w e r s , w h e r e o f t h i s i s h o l d e n t o be a k i n d e , w h i c h h a t h
mo ve d me t o p r e f e r i t u n t o t h e f i r s t p l a c e .
He s a i t h ,
t h a t t h e j u i c e m i x e d w i t h some u n c t i o n s o r o i l i e t h i n g ,
and b o i l e d t o t h e form of a l y n i m e n t , h e l p e t h t h e c h o p p e s
or r i f t e s of t he fundament,.
The h e r b b o i l e d w i t h w h i t e
w i n e , h o n i e , a nd a l i t t l e a l l o m , d o t h c u r e h o t u l c e r s ,
and c a n k e r s o f t h e m o u t h .
After
tific

the

seventeenth

and t he h e r b a l

c e n t u r y b o t a n y b e c a me mo r e

wa s g r a d u a l l y

r e p l a c e d by t h e

scien­

flora

and

the ph a rm aco p o eia.
3.

Mo d e r n D e v e l o p m e n t s

I n 19 1 0 w i t h P a u l
arsenical

Salvarsan

Ehrlich's

used

in the

s y n t h e s i s of
treatment

t h e m o d e r n e r a o f c h e m o t h e r a p e u t i c s wa s
mochin,

the

synthetic

modern s u l f a

drugs,

sulfadiazine

gave hope

fight

any d i s e a s e

less

into

the

organic

syphilis

inaugurated.

(27),
Plas-

a n t i m a l a r i a l , Domagk' s P r o n t o s i l ,

such as

with

c o n s i d e r a t ion of p l a n t s
or

of

in Che mo t he ra p y

that

sulfanilamide,
one d a y

a synthetic
as

s u l f a t h i a z o 1e , a n d

i t w o u l d be p o s s i b l e
drug.

suppliers

a nd

Consequently,

to
the

o f m e d i c i n a l s w e n t mo r e

t h e b a c k g r o u n d and A l e x a n d e r

Fleming's

observation

6

i n 1929
the

(30)

that

t h e m o l d P e n i c i l l ium n o t a t urn c o u l d

g ro w t h o f M i c r o c o c c u s pyo gene s v a r . a u r e u s

unnoticed.

About

investigated
taining

the

ten years
same

antibiotic

Consequently,
importance

191+0 t h e

Florey

agent

have

organisms

as

suppliers

like

s h o we d g r e a t p r o m i s e

Since

one

organisms,

is prone

i t ma y b e

to

surprising

of d i f f e r e n t

species

examined f o r

antibacterial

the r e s u l t s

of

the

researches

Insofar

common t h e r a p e u t i c

antibiotics,

since

microorganisms,

it

only

appeared

used t h e r a p e u ­
as p r o m is in g .

a n tib io tic s with m icro­
that

a large

n u mb e r

I t may b e c o n c e d e d

dramatic

that

in h i g h e r p l a n t s

success

that

has a t ­

on m i c r o o r g a n i s m s .

has yet

interest

a g e n t s was a n ­

a u r s o m y c i n a nd

and a r e

antibiotics

a s c a n be d e t e r m i n e d ,

from a seed p l a n t

the

the

and a f t e r

of h ig h e r p l a n t s have been

activity.

search for

activity

group r e - e s t a b l i s h e d

of m e d i c in a ls

to fin d

and v a r i e t i e s

as y e t met w i t h

tended the

associate

in o b ­

(31).

streptomycin,

t i c a l l y w h e r e a s ma n y o t h e r s h a v e n o t

creased

succeeded

o f a g r e a t ma n y a n t i b i o t i c

Some o f t h e s e ,

terramycin,

its

a nd C h a i n a t O x f o r d

and

t h e work of t h e Ox f or d

of p l a n t s

isolation

nounced.

have not

almost passed

c r u d e p r e p a r a t i o n s w h i c h had r e m a r k a b l y h i g h

a g a i n s t ma n y G r a m - p o s i t i v e

the

later,

inhibit

s hown s u f f i c i e n t

use.

Nevertheless,

in h i g h e r

plants

is n o tic e d
a few of

a c t u a l l y have been u s e f u l

no a n t i b i o t i c

that

isolated

promise
this

to w a rra n t

has not

de­

as p o s s i b l e

sources of

even

case

in t h e

t h e ma n y a n t i b i o t i c s

therapeutically.

of

isolated

7
Ip. P l a n t s
Several

Possessing A ntibacterial

surveys over

higher

a c t i v i t y have been p u b l i s h e d
i n 192-1-3 ( 6 7 ) .
tions

are

in

(Ip, 14.5 ) ,

a nd W i l l e k e
a complete
are

reports

a n d Ma c D o n a l d
(92).

having

a n d l p3. 2 p e r

of a n t i b a c t e r i a l
is w ith in

to

bacterial

investiga­

a g a i n s t M.
cent

of

the

select

plants

Weatherwax,

they

carried

c a n be

that

and

that

several places
systematic

in h i g h e r p l a n t s .

seen from the

(1^0,55,32) .

plants

activity

a mo r e

first

I t was f o u n d

t u b e r c u lo s is o ccu rred
tested

represent

indicate

on a t

anticipate

and W i n t e r

do n o t

antibacterial
but

in C a l i ­

(13),

references

principles

reach,

This demonstrates

possible

these

in Canada

significance;

Cne c a n t h e r e f o r e

goal

activity

in 16 .8 ,

in t h r e e

consecutive

through experience

which p r o b a b l y w i l l

21.1,

it

exhibit

is
anti­

activity.

Besides,

i t may be c o n c l u d e d f r o m t h e

antibacterial

flowers

These

o f L u c a s and c o - w o r k e r s

surveys.

of

wa s b y O s b o r n

H u g h e s a nd a s s o c i a t e s

screening programs are

determination

that

Bishop

( 7l p) ,

only of p r e l i m i n a r y

this

of which the f i r s t

a nd c o - w o r k e r s a t M i c h i g a n S t a t e

s urve y of p l a n t s

in t h e w o r l d .

that

Indiana

i n G e r ma n y

very active

That

of Lucas

antibacterial

( 3 2 , 3 3 , 3]p, 3 5 , IpO, Ip1, £ip, 5 5 , 56 ) , S a n d e r s ,

a nd Mc Cl u n g
fornia

having

Among t h e m o s t e x h a u s t i v e

those

University

plants

A ctivity

and

leaves

activity

o c c u r s most f r e q u e n t l y

a nd

frequently

less

R o o t s have n o t been examined to
conclusions

survey r e s u l t s

c a n be d r a w n .

in th e

such an e x t e n t

in the

fruits
that

and

seeds.

similar

8

Phenolic

and a c i d i c

of b a c t e r i o l o g y
exhibit

it

has been recognized

antibacterial

activity.

i n Th ymu s v u l g a r i s .
isolated
Link

Others

Th e e a r l i e r

and

woods o f P i n u s

because

compounds.

acid

(19)

are

i t s monomethyl

Lac t o n e s . - L a c t o n e s a s

the

roots

cially

The

ether

moderate

isolated

from the

aucupar ia,

ripe

(86).

against

berries

of

example of

of

found

to have

(1|_2),

prop­

carvacrol

(78)

from the h e a r t inhibited

the

fungi.
seem t o e x h i b i t

lactone

kawain

s howe d

several

antibacte­

isolated

such a c t i v i t y ,

activity

bacteria;

from
espe­

for

So r b u s

antibacterial

(17)-

responsible

c a u s e d by e a t i n g
disease

acid

lactone

a substance

this

the

Parasorbic

an a n t i b a c t e r i a l

cattle

against

but

the mountain ash,

( 18 ) i s o l a t e d

disease

of c e r t a i n

P r o t o a n e m o n i n a nd a n e m o n i n f r o m

The c o mp o u n d r e s p o n s i b l e

whi c h was a l s o

and c a t e c h o l

such s u b s t a n c e s .

s ho we d c o n s i d e r a b l e

and L i n k

the hem orrhagic

clover.

present

antibacterial

acid

c o m p o u n d s wa s f a i r l y h i g h .

is another

I n 19l fi C a m p b e l l
for

(17)

activity

t o x i c i t y of both

substances

A 1 1 ium c e p a , b y

isolated

a group

unsaturated

Gonococcus

An e mo ne p u l s a t i l i a
fungi,

the

examples of

o f P i pe r me t h y s t i cum ( I l f )

against

of

s y 1v e s t r i s b y R e n n e r f e l d t

properties.

acid

products

Thujic

g r o w t h o f a n u mb e r o f w o o d - r o t t i n g

rial

days

is thymol,

are p r o t o c a t e c h u i c

interest

o f many o f t h e

Pinosylvine

early

phenolic

One o f t h e s e

work on s t e a m - v o l a t i l e

and c h ami c

the

(58,2).

gymnos per ms p r o v o k e d

(19),

that

from a pigmented v a r i e t y of onion,

and c o - w o r k e r s

erties

su b s ta n c e s. - Since

sweet

wa s d i c u m a r o l ,

properties

(17)-

9
Alkalo i d s . - Since
macological
has been

properties,

given

hyoscine,

of b a c t e r i a

their

that

This confirms

bacterial

antibacterial

and c o c a i n e

they all

the

action

atropine,

against

a n u mb e r

s h o we d o n l y n e g l i g i b l e

finding

that

a s A t r o p a a nd D a t u r a e x h i b i t

activity.

interesting phar­

Ha i m ( Lp3) t e s t e d

morphine,

a nd f o u n d

drug p l a n t s

possible

some a t t e n t i o n .

aconitine,

activity.

alkaloids possess

such w e ll- kn ow n
no d e t e c t a b l e

Lamb i n a n d B e r n h a r d

(51)

several

alkaloids,

such as apomorphine,

strychnine

against

activity

appreciable.
B e r s c h a nd Dopp ( 9 ) on t h e o t h e r
rt
t h a t c e p h a r a t i n e , b e r b e r i n e , a nd s a n g u m a r i n e a l l

M.

were f a i r l y

t ube r c u 1o s i s a n d f o u n d

active

a g a i n s t M.

Lamb i n a n d B e r n h a r d
concentration

if/ m l .

M.

t u b e r c u l o s i s , whereas Bersch
a concentration

effect.

These

the

results

are

influenced

detecting

examples

to

of

only

sim ilarly their

10 t o 1+0

testing.
extent

a nd Dopp

tu b e r c u lo s i s at

isolated

several

a n d Dopp

antibacterial

by B e r s c h

that

berberine

the

growth of

inhibited

( 1 oc . c i t . ) m e n ­
if/ml.

s how how d i f f i c u l t

such a g r e a t

tested

g r o w t h o f M.
I t wa s

150

of b a c t e r i o l o g i c a l

a nd m e a s u r i n g

sanguinarine

against

of

( 1oc . c i t . ) s t a t e d

the

ml .

and

tuberculosis.

at

the

conessine,

not

hand f o un d

tioned

berberine,

tested

anti­

it

for

the

i s t o compare

The r e s u l t s

obtained

by the methods

used f o r

activity.

The a l k a l o i d

(lo c . c i t . ) inhibited

c o n c e n t r a t ions 0 . 5 to

b y J o h n s o n et_ aj^.

same

5

(1+7) a n d f o u n d a c t i v e

s p e c i e s o f E s c h e r i c h i a and A e r o b a c t e r .

10
The

steroidal

Fontaine

a n d Ma ( 5 7 )

pyogene s v a r .
E.

alkaloid

co1i .

aureus

On t h e

toward c e r t a i n

glycoside,

tomatine,

reported

s h o we d

l ow a c t i v i t y

a nd B.

s u b t i 1 i s a n d no a c t i v i t y

other

hand,

it

by

against Micrococcus

exhibited

against

a high a c t i v i t y

fungi.

B . C h e m i s t r y a n d P h a r m a c o l o g y o f S o l a n u m P s e u d o c a p s i c um L .
So 1 anurn p s e u d o c a p s i c u m L.
Cherry,

C h r i s t m a s C h e r r y , or N a t a l

branching
feet.

leafy

shrub t h a t

It has narrow,

lanceolate

a shiny surface.

s o l i t a r y or

few

fruit

in l a t e r a l

makes

it

probably a native
Madeira off
tributed

the

northwest

ornamental

i n 1929
cent

toxic

clusters.

(15) that

(50)

the

identical

perhaps

regions

It

"Probably physalien

to

tomatoes

the
It

green

flowers
orange

island

are

cherry­
is

of

is widely d i s ­

of A f r i c a
g r o wn

a nd

in g r e e n ­

in f i e l d s

are

the b e r r i e s

cultivated.

contain

the

Br e y e r - B r a n d w i j k r e p o r t ­

leaves contain 0.25

sola ine.

four

The p l a n t

is w ide ly

non-volatile
with

bright

a n d c a n be g r o wn

that

dried

of th ree

ornamental.

and p h y s a l i n . " *

o f a n u n k n o wn ,

and n o t

Its

subtropical

purposes

is a small

and w h i t e

of A f r i c a .

in F l o r i d a .

i n 1927

pigments phyllorhodin

per

small

in t h e U n i t e d S t a t e s where

Kyi i n f o u n d

ed

and

Jerusalem

l e a v e s which are dark

The

coast

(66),

a height

of t h e Old Wo r l d ,

has been n a t u r a l i z e d

anywhere

Cherry

an a t t r a c t i v e

in t r o p i c a l

houses for

c o mmo n l y c a l l e d

g r o ws t o

and p o s s e s s

like

(5),

alkaloid,

- 0.53

wh i c h was

11

Watt

e_t_ al_.

loid product
circulatory
called

from the
system.

leaves

the

action

in t h e i r

irritation

application
of

alkaloidal

injected

concentrations

the

heart

a c t i o n by p r o d u c i n g

wa s p o s s i b l e

to

Its

administer
toxicity

alka­

The a l k a l o i d
stomach,

subcutaneously.

toxic

block,

the

p r e p a r a t i o n wa s

second p a p e r .

of

aur i c u l o v e n t r i c u l a r

subjects.

wh e n

of

o f S o l anurn p s e u d o c a p s i cum on t h e

p r o d u c e s v o m i t i n g wh e n g i v e n b y t h e

local

ization

described

The a m o r p h o u s

s o 1a n o c a p s i n e

irritant,
gives

(87,88,89)

resulted

and
The

in wide

sinus

disorgan­

arrhythmia,

and w e a k e n i n g of

the muscle.

d o s e s of

0 . 0 8 1 | g.

is about

0.06 to

equal

is

to t h a t

It

t o huma n

of c o c a i n e

h y d r o c h 1o r i d e .
Muenscher
the

alkaloid

stated

possess

toxic

Schnell

that

the b e r r i e s

substances

that
(82)

antibacterial
inactive

the p l a n t ,
solanine

may c a u s e
tested

to

and s o l a n i d i n e .

s h o u l d n e v e r be e a t e n

activity

in_ v i t r o

in a d d i t i o n

as t h e y

severe p o iso n in g .

the b e r r i e s
and f o u n d

o f S_. p s e u d o -

that

an a q u e o u s

a g a i n s t Micrococcus pyogene s

co1i .

Lucas et
o f S.

that

and T h a y e r

capsicum for

and E.

mentioned

solanocapsine, contained

He a l s o

e x t r a c t was

(66)

a l.

found

( 32)

that

a water

p se u d o c ap s i c um wa s p r o m i s i n g l y

extract

active

against

t u b e r c u l o s i s and M i c r o c o c c u s pyogene s , w h e r e a s
extract

of

pyogenes.

the

leaves

The

alkaloid

characterized

s h o we d a c t i v i t y

by B a r g e r

of

the

roots

b o t h M.

an e t h a n o l i c

only a g a in s t Micrococcus

so 1 a n o c a p s i n e w a s f i r s t
a nd F r a e n k e 1 - C o n r a t

(6),

chemically
who t h o u g h t

12
that

t wo a l k a l o i d s w e r e p r e s e n t .

( 78 ) f o und o n l y o n e

alkaloid,

C . Chemical

Schlittler

and U e h l i n g e r

n a m e l y so 1 a n o c a p s i n e .

Studies

on So 1a n o c a p s i n e

I n 1929 B r e y e r - B r a n d w i j k p r e p a r e d f r o m t h e
S o l a n n m p s e u d o c a p s i c um a n a m o r p h o u s
which had a d e p r e s s a n t
In 1936 B a r g e r
leaves

of

assigned

the
the

s i d e was

this

pentoses.

such

c o mp o u n d

acid

The

the

isolated

from the

a l k a l o i d which t h e y

t wo w o r k e r s o b t a i n e d

but

the

crude

gave r e a c t i o n

authors

No a l k a l o i d

alkaloid
for

conceived

formed d u r in g

glyco­

trea te d with 2 N

carbohydrates

that

an a m o r ­

f o r m u l a C2 6 H^ 2 N2°1|

so 1a n o c a p s i d i n e .

the

and f o r

t wo b a s e s w e r e

isolation

and n o t p r e s e n t

in the p l a n t .

So 1a n o c a p s i n e w a s u s e d f o r
reactions,

b u t due

hydrogenation,
abundant
in t h e

(6)

w h i c h was f o u n d t o h a v e t h e

secondary products
as

a crystalline

Furthermore,

isolated;

hydrochloric

(15),

f o r m u l a Cp^Hj ^ NpQp o r ^ 2 ^ 1^2^ 2 ^ 2 anC^ nam0<^ t h i s

alkaloid

and c a l l e d

product

of

on t h e h e a r t .

a n d F r a e n k e 1- C o n r a t

same p l a n t

so 1a n o c a p s i n e .
phous

effect

alkaloid

leaves

this

to

so 1a n o c a p s i d i n e .

to

t wo e q u i v a l e n t s

of n i t r o u s

so 1a n o c a p s i n e .

group,

out

for

selenium de­

o n t h e mo r e

Assuming a s i m i l a r i t y of

they applied

dehydrogenation

a nitroso

of m il d d e g r a d a t i o n

insufficient material

r e a c t i o n was c a r r i e d

t wo a l k a l o i d s ,

acquired

a n u mb e r

acid;

the

results

of

So 1a n o c a p s i n e

structure

selenium
reacted with

a s e c o n d a r y a mi n o g r o u p

a p r i m a r y a mi n o g r o u p wa s c o n v e r t e d

13
into

a hydroxyl

group,

and p r o b a b l y t h e

p r e s e n t was e l i m i n a t e d
According

s h o we d t h r e e

four

higher

gen

at

(or

active

temperature,

accounts

two)

of

duced t h a t
with

for

the

one

three

a hydroxyl

acetic

Since

that

wa s

of

a c o mp o u n d t h a t

the

acetone

acetyl

these

this

workers

acetyl

ocapsidine,
threne),

second h y d r o ­

group fo r

hydrogens,

that

reported

t h e hydroxyl

the

second

that

it

ring.
a n d F r a e n k e 1- C o n r a t

s h o we d o n l y o n e a c t i v e
complex r e s u l t e d

acetone

hydrogen.

in a m o n o a c e t y l

attached
split

to
off

the

(6)

to y i e l d
Acetylation

so 1a n o c a p s i n e ;

s e c o n d a r y a mi n o

during

isolation

By s e l e n i u m d e h y d r o g e n a t i o n o f

Diels hydrocarbon

diacetyl

they concluded

i t was assumed

c o u l d be c o n d e n s e d w i t h

derivative.

On h e a t i n g

i t wa s r e a d i l y e l i m ­

the f u n c t i o n of

by Barger

a c e t o n e p a r t was

that

a double bond,

Inasmuch as

one

i t wa s d e ­

a neutral

( i b i d . ) found

and b e l i e v e d

group was assumed

group and t h e

the

and

s e c o n d a r y a mi n o

s o l a n o c a p s ine y i e l d e d

n o t be d e t e r m i n e d ,

solanocapsine

r o om t e m p e r a t u r e

the

active

a heterocyclic

I t wa s a l s o

the

four)

( ibid. ) solano-

g r o u p wa s o r i g i n a l l y p r e s e n t .

i t wa s t e r t i a r y .

a member

that

Since

as w a t e r w i t h f o r m a t i o n of

oxygen atom co u ld

at

the f o u r t h being

group.

(or

group d i d not' a c e t y l a t e
inated

hydrogens

a n d t h e p r i m a r y a mi n o

anhydride,

derivative.

of

and F r a e n k e 1- C o n r a t

i n t h e p r i m a r y a mi n o

group

originally

as w a t e r .

to Barger

capsine

hydroxyl

of

solan-

( 3 1- m e t h y l - e y e 1o p e n t e n o p h e n a n -

2 - m e t h y l - 5 - e t h y l p y r i d i n e , a nd l | - m e t h y 1 - 2 - e t h y l p y -

r i d i n e were

obtained.

Based

upon t h e s e

results

the

authors

11).
proposed
n u mb e r

for

so 1a n o c a p s i n e

as p re s e n te d

19lf5> R o c h e l m e y e r

( ?L|_) s u g g e s t e d

so 1 a n o c a p s i n e w i t h 27 c a r b o n s ,

capsine

in

Solanum.

line

with

Without

formula given

other

a str uctural

formula

w h i c h would b r i n g

solano-

alkaloids

any r e p o r t e d

isolated

experimental

i n d r a w i n g n u mb e r

evidence

the

ch5

HC

CH-

L

OH

"NH

2
I

Schlittler

and U e h l i n g e r

s t r u c t ure, o f

(78)

sol anocaps i n e .

i d e n t i t y of t h e i r

c o mp o u n d w i t h

(6);

formula

but

wa s c l a s s i f i e d
solanidine,

of

the

published,in

compound.

the base

isolated

that

and Fr a e n k e 1- C o n r a t

By t h i s

solasodine,

(op_.

the

in agreement w i t h
formula

a s a n a g i y c o n e w i t h 27 c a r b o n s

tomatidine,

1952? a paper

T h e r e w a s no d o u b t

they decided

wa s mo r e

analysis

Vk

"H 2

11

and F r a e n k e l - C o n r a t

tary

genus

I I wa s s u g g e s t e d .

HO

the

from the

CH

LH

on t h e

in drawing

I.

In
for

a structure

et

about
by B a r g e r

empirical
their

elemen­

so 1a n o c a p s i n e

( o th e r s being

cetera) .

c it . ) they maintained

Like Barger
that

the

c om­

pound had

a p r i m a r y and a s e c o n d a r y a mi n o g r o u p

a nd f o r m e d

trimethyl

derivative.

one of

o x y g e n a t o m s wa s e i t h e r

They a l s o
present

concluded

that

as a t e r t i a r y

hydroxyl

a

the
group

1?
or

as a n o n r e a c t i v e

oxygen was p r e s e n t
Schlittler
could not
alkali.

be

secondary hydroxyl

group,

while

the

second

a s an e t h e r .

and U e h l i n g e r

acetylated

or

(6)

found

eliminated

By d e h y d r o g e n a t i o n o f

that

t h e h y d r o x y l group

as w ater

by a l c o h o l i c

so 1a n o c a p s i n e w i t h

selenium,

D i e l s h y d r o c a r b o n was o b t a i n e d b e s i d e s B - e t h y l -%-met h y l p y r i dine.

Short

a basic
tained

time h e a tin g

reacting
all

the

of

so 1a n o c a p s i n e w i t h

c o mp o u n d w i t h

carbon atoms,

the

into

the

dicate

compound.

that

The
c h l o r ide

ultraviolet

spectrum of

s h o we d a mo r e

under

sults

the

hydroxyl

given

in d r a w i n g number

influence

authors proposed

the hydroxyl

Ill

introduced
in­

s o 1a n o c a p s i n e h y d r o band t h a n the

i s why i t wa s
of a c i d .

a structure

III.

con­

and t h e

s p e c t r u m s e e me d t o

trimettyl

intense

It

conjugated.

so 1 a n o c a p s i n e , a n d t h a t

r i n g E opened

lost

d o u bl e bond were

d o u b l e bo nds were

infrared

of t r i m e t y l
that

the

The

f o r m u l a C^yH^NO.

but had

p r i m a r y a mi n o g r o u p s a n d t h r e e

s e l e n i u m gave

for

spectrum

suggested

Fr o m t h e s e
so 1a n o c a p s i n e

re­
as

III.

EXPERIMENTAL

A.

A p p a r a t us

Wiley l a b o r a to r y m i l l ,
knife

cutter

A r t h u r H.

with

No.

1 . - A small

interchangeable

Th o ma s Co mp a n y ,

sieve

Philadelphia,

F l a s h e v a p o r a t o r . - The r o t a t i n g
by C r a i g

and G r e g o r y

(22)

and b u i l t

Kedzie Chemical L a b o r a t o r y .
pressure

and

a temperature

pH-meter. operated.

with the

the

Sold by

described

shops of
at

the

17 t o 20 mm.

below 50°C.

c a p i l l a r y method.

capillary.

in the

glasselectrode,

line

i n m a k i n g pH m e a s u r e m e n t s .

a p p a ra tu s . - A mineral

a mechanical

rotary

Pennsylvania.

I t was o p e r a t e d

pH m e t e r w a s u s e d

beaker with

openings.

evaporator

A Be c k ma n mo d e l H 2 ,

Melting point

scale

The

stirrer

liquid

oil

b a t h was used

bath consisted

and t h e r m o m e t e r

M e l t i n g p o i n t s were

of a

supporting

reported without

correc-

t ion.
C h r o m a t o g r a p h y c o l u m n s . - Two t y p e s o f c o l u m n s w e r e
The f i r s t
reduced

t y p e wa s

to

a small

a straight
diameter

glass

effluent

a p l u g o f P y r e x g l a s s wo o l wa s
outlet
smaller

tube
than

an e v en b a s e

and c o v e r e d w i t h
the

inside

for the

column
outlet

inserted

a filter

diam eter of

adsorbent.
16

(50 x 2 . 1

into

paper

( 0 . 5 cm.
the
disk

cm.

used.
i.d.)

i.d.).

top of the
slightly

thecolumn.T his p r o v i d e d

17

with

The

second

the

lower

The f l o w r a t e
the

effluent

paper

t y p e was

end c o m p r i s e d by a ground
of

the

A p l u g of Pyrex

inserted

as d e s c r i b e d

In o r d e r

to

saturate

cylinder.

plate

that

solvent.

the

paper

cylinder

When r e a d y f o r

m a g n e t was r e mo v e d and t h e
the

trophotometer,

and a f i l t e r

as d e s c r i b e d

side

solvent
to

the

(23).

a piece

other

of

of the

on t o p o f t h e

glass

i r o n w i r e wa s h e l d b y t h e

i n t h i s wa y k e p t
development

of

above

the

level

spots,

of

the

bottom of the paper

cylinder

lowered

spectrophotometer, - A recording

infrared

spec­

model

21,

Connecticut,

U ltraviolet
trophotometer,
rated,

at

solvent.

Infrared

Norwalk,

by a stopcock

p a p e r wa s u s e d

the

( ^ 29/ 1+2) .

above.

a small magnet

cylinder,

I.d.)

joint

g l a s s wo o l

the paper with

By p l a c i n g

covered

ma g n e t and t h e

into

1 filter

i r o n w i r e wa s h o o k e d f r o m one

paper

the

glass

c h r o m a t o g r a p h y . - The a s c e n d i n g m e t h o d

b y C r a m e r w i t h Wh a t ma n No.

bent

( 6 0 x 2 . 2 cm.

e l u e n t was c o n t r o l l e d

outlet.

d i s k were
Paper

a g l a s s column

f r o m The P e r k i n - E l m e r

Corporation,

wa s u s e d .

spectrophotometer. - A ratio

recording

m o d e l D K j ^ f r o m B e c k ma n I n s t r u m e n t s ,

South Pasadena,

spec­

Incorpo­

C a l i f o r n i a wa s u s e d .

Visible

s p e c t r o p h o t o m e t e r . - A m anually o p e r a t e d Bausch
<i
S j* t c f f r « H t c Z . o
a n d Lomb s p e c t r o p h o t o m e t e r ^ w a s u s e d f o r m e a s u r i n g o p t i c a l
density

in the

visible

r e g i o n of the

P o l a r i m e t e r . - The o p t i c a l
polarimeter
Caldwell,

spectrum.

r o t a t i o n wa s m e a s u r e d

in a

m a n u f a c t u r e d a n d s o l d b y O. C* R u d o l p h a n d S o n s ,

New J e r s e y .

18
Barnstead
on t h e

e x t r a c t o r . - A large

principle

with p eriodic
fresh

siphoning

liquid.

C o mp a n y ,

of c o m p l e t e
off

Fabricated

Incorporated,

immersion of
of

solvent

by B a r n s t e a d

Forest H ills,

a nd u s e d t h r o u g h c o u r t e s y o f D r .
Agricultural

H.

solid

working

in th e

liquid

and r e p l a c e m e n t
Still

and S t e r i l i z e r

Boston,
Sell,

by

Massachusetts

Department

of

Chemistry.

Percolator
which

capacity extractor

instead

porous disk

type

of

e x t r a c t o r . - A Soxhlet

type

of e x t r a c t o r

a s i p h o n i n g d e v i c e wa s f u r n i s h e d w i t h

allowing

the

t h r o u g h c o u r t e s y o f Dr.

extract
H.

to pass

Sell,

through.

Department

a

Used

of A g r i c u l t u r a l

Chem i s t r y .
Hydrogen c h l o r i d e
generated

by d r o p p i n g

aratory funnel

concentrated

(connected

sodium c h l o r i d e .
concentrated

g e n e r a t o r . - Hydrogen c h l o r i d e

to

B. Reagents,

M aterials

Diethyl
washed w i t h
chloride

- All

grade

by p a s s i n g

and A n a l y t i c a l

it

solid

through

a

chemicals

or C . P .

Methods

and M a t e r i a l s
used f o r

analytical

purposes

quality.

e t h e r . - T h i s w a s o b t a i n e d f r o m M e r c k a n d Co mp a n y ,
a s o l u t i o n of f e r r o u s

and m a g n e s i u m s u l f a t e .

sodium and

) onto

acid washer.

1. R e a g e n t s

were r e a g e n t

acid from a sep­

a suction flask

The g a s wa s d r i e d

sulfuric

Chemicals.

sulfuric

wa s

stored

over

sulfate,
Finally

sodium w i r e .

dried

over

calcium

i t was d i s t i l l e d

over

19
A1 u m i n a . - A l c o a
I.”

alumina, grade F-20,

When d i s p e r s e d

reacted

to produce

i n a s m a l l a mo u n t o f w a t e r ,
a pH o f

around

chromatographic
*~>h- C S t r e e t ,

cals,

analysis.
Elmont,

Dowex 1 . - A n i o n
a nd

12 p e r

Co mp a n y ,

cent

cal

a nd I4. p e r
Co mp a n y ,

Lon g

Island,

exchange r e s i n

cent

cross

Midland,

acid

alumina

activity

grade

I

New Y o r k .

o f 50 t o
from

100 m e s h s i z e
Dow C h e m i c a l

Michigan.

to

resin

linkage.

of

100 t o 2 0 0 me s h

Obtained

f r o m Dow C h e m i ­

Michigan.

0 . 0 2 N HC1 . - P r e p a r e d
hydrochloric

this

O b t a i n e d from Al upharm Chemi­

Dowex 5 0 ■ - C a t i o n e x c h a n g e
size

( Wo e i m) ;

cro sslinkage. Obtained

Midland,

"activity

1 2.

Ac i d a l u m i n a . - A l u m i n u m o x i d e
for

80 mesh,

by d i l u t i n g

1 liter.

3 m l . of

Standardized

concentrated

against

borax,

Na 2 B ^ 0 7 , 1 0 H2 0 .
0 . 0 2 N NaOH.

- Twenty-five m i l l i l i t e r s

d r o x i d e was d i l u t e d

to

1 liter

with f r e s h ly boiled water.

N s o d i u m h y d r o x i d e wa s p r e p a r e d f r o m 50 p e r
droxide.

Standardized

Tomatine.
fer,

against

- Obtained

by t h e p r o c e d u r e

cent

sodium h y ­

0.02 N h y d ro ch lo ric

t h r o u g h c o u r t e s y o f Dr.

E a s t e r n U t i l i z a t i o n Research Branch,

prepared

of N sodium h y ­

of

Fontaine

Molisch*s r e a g e n t . - A 5 per

cent

et

acid.
P.

U.S.D.A.
al .

S.

Schaf­

a nd may be

(31a) .

ethanolic

s o l u t i o n of

<X. - n a p h t o 1 .
Ma y e r * s r e a g e n t . solved

i n 60 m l .

1 . 3 6 g.

of water

of m ercuric

c h l o r i d e wa s d i s ­

and 5 9 * of p o t a s s i u m

iodide

in

20

10 m l .
to

of w a te r.

100 m l . w i t h w a t e r

rides

of most

s h o u l d be
dilute

small

added to

the

in v e r y d i l u t e

sulfuric

contain

acetic

or

A few d ro p sof
of

some

1 . 6 6 g.

acid

acid

of water

sprayed w ith

acid with

a n d mo r e

than a

precipitate

should

a lk a lo i d s are

be added a s

soluble

in e x c e s s

olive-brown

and 0 . 9 3

th is mixture

spots

of

standing
crystal
this

acetic

overnight
deposit

decantate
use as

of

of a n i l i n e

reagent.

wa s a d d e d t o

for

10 m i n u t e s .

and o t h e r

g.

ofsodium

a c i d were b o i l e d
the

- A mixture

for

sugars

sodium a c e t a t e .

was a d d e d t o 80 ml .

ethyl

of 2 . 6

iodide,

For

acetate.

the m ixture.

g.

spot

While

glacial

from the

acetic

development
of g l a c i a l
shaking

ni l .

After

Twenty m i l l i l i t e r s
of

of

a nd 2 5

a few m i n u t e s .

s u p e r n a t a n t was d e c a n t e d

s t o c k r e a g e n t was mi x e d w i t h 50 m l .
of

The p a p e r wa s

105°C.

spots

sugars

wa s d i s s o l v e d

1- b u t a n o l .

a nd wa r me d t o

7-0

stock reagent.

120 m l .

reducing

(23).

bismuth carbonate,

glacial

g.

reddish-brown

Modified Dragendorff
basic

r e a g e n t . - For

saturated with

Aldopentoses produce

a nd

The r e a g e n t

The a l k a l o i d

alkaloid

the reagent

hydrogen p h t h a l a t e

phthalic

i n 100 m l .

for

hydrochlo­

distinctly
acid.

the

the

reagent.
Aniline

of

else

and d i l u t e d

solution.

rendered

a c i d or

amount of e t h a n o l

the p r e c i p i t a t e s

It p r e c i p i t a t e s

solutions

should not

soluble.

s o l u t i o n s were mixed

(2LjJ.

alkaloids

hydrochloric

solution

is

The t wo

acid

20 m l .
acetic

10 m l .

of

of
acid

of water

21
Paper
a nd

it

c a n be

does not

dipped

spread

in t h i s

the

solution for

developed

colors

B u f f e r s . - The b u f f e r s w e r e p r e p a r e d
t a b l e s by G o r t n e r
Anhydrous

of

the

used f o r

the procedure

crude m a t e r i a l . -

g r o wn

first

conducted

antibacterial

over

an Op p e n a u e r

o f F u c h s and

For

in t h e f i e l d .

frost

and

in o r d e r

forced

draft

reason

to o b tain

stored

under

refrigeration.

the

stability

I t wa s f o u n d t h a t

all

the m a te ria l

plant material

a maxi mal

temperature

and r o o t s wer e

separated

before

All

plant

the

just
Tests

of

the

a c t i v i t y wa s

wa s r e f r i g e r a t e d

oven at

separately.

sufficient

These were h a r v e s t e d

to determine

substance.

this

In o r d e r

o f S_. p s e u d o c a p s i c um a b o u t 5 0 0

re g a rd le ss whether

dried.

parts

followed

uniform q u a li t y

specimens were

retained

the

(36).

Source of

were

to

sulfate.

o x i d a t i o n was p r e p a r e d

before

according

(39).

A l u m i n u m p h e n o x i d e . - The r e a g e n t

material

(8 5 ).

a c e t o n e . - Th e a c e t o n e wa s d i s t i l l e d

anhydrous calcium

Reich s te in

development

wa s d r i e d

in a

at

Green

drying

material

was g r o u n d

Analytical

Procedures

60°C.

and l a t e r

or

stored

in a W ile y m i l l

2 0 me s h .

2.
D e te r m in a t io n of

acetyl

g r o u p s . - The n u mb e r o f

g r o u p s was d e t e r m i n e d by t h e p r o c e d u r e
described

by M i l t o n

and W a t e r s

(60).

acetyl

o f Kuhn and R o t h a s
N ethanolic

potassium

to

22

h y d r o x i d e was use d f o r
triacetyl

hydrolysis

except

in the

s o 1 a n o c a p s i n e , w h e r e N 1- b u t a n o l i c

i d e wa s u s e d .

The

latter

more e f f i c i e n t

in t h e

reagent

hydrolysis

c o mp o u n d wa s d i s s o l v e d

case

in p y r i d i n e

before

the

potassium hydrox­

has been regarded
of N - a c e t y l

of

t o be

compounds.

addition

The

of hydro-

l y t ic r e a g e n t .
The m e t h o d u s e d b y A l i c i n o
mination

of O -acetyl

bert

(7).

stirrer

weight

according

A beaker

served

deter­

groups.

R a s t 1s molecular
wa s d e t e r m i n e d

(1) was e mp l o y e d f o r

to

d e te r m i n a ti o n . - Molecular weight
the procedure

of mineral

as the h e a t i n g

oil
bath.

of Belcher

a n d Go d -

o u t f i t t e d with mechanical
A thermometer

calibrated

f r o m 9 0 ° t o 2 0 0 ° C . w i t h 0 . 2 ° s u b d i v i s i o n s was used f o r
atu re measurements.
Determination
procedure
were

Twice

sublimed

of methyl

and r e a g e n t s

c a mp h o r wa s u s e d

groups a t t a c h e d

temper­

as

solvent.

t o c a r b o n . - The

d e s c r i b e d by M i l t o n and W a t e r s

( 6 1)

adopted.
Neutralization

e q u i v a l e n t . - A weighed

c a p s i n e wa s d i s s o l v e d

in e t h a n o l

of 0 .0 1 0 2 N h y d r o c h l o r i c

acid,

0.0120 N potassium hydroxide.
neutralization
during

the

Fr om t h e

titration

to Clark

solano-

a known e x c e s s

a nd t h e n b a c k - t i t r a t e d
data obtained,

e q u i v a l e n t was c a l c u l a t e d .

with
the

The pH c h a n g e

wa s f o l l o w e d w i t h a Be c k ma n p H - m e t e r .

Semi-micro K jeldahl
according

containing

sample of

a n a l y s i s . - Procedure

(21) were f o l l o w e d .

a nd r e a g e n t s

Digestion

and

distillation

equipment

used

laboratory.

in t h i s

wa s n e c e s s a r y

to

Van S l y k e

insure

a mi n o

b a s e d on m e a s u r e m e n t
RNH2
During

the

for

+

a n a l y s i s wa s t h a t
A digestion

complete

nitrogen

and S a n d s t r o m

gas

liberated

ROH

+

excess n itro u s

least

N2

in the
+

6 hours

acid

HN0 3

+

2 NO

and r e a g e n t s were f o l l o w e d

is

reaction

H2 0

decomposes

in the f o r m a t i o n of n i t r i c

3 HNC>2 ------------->
Procedure

at

digestion.

HN0 2 ------------ »

spontaneously r e s u ltin g

time of

a nd

d e t e r m i n a t i o n . - The m e t h o d

of n i tr o g e n

reaction,

as m o d if ie d

+

oxide:

H2 0.

according

t o Mo r r o w

(63).

E l e m e n t a r y a n a l y s i s . - The a n a l y s e s w e r e p e r f o r m e d b y
either

Huffman M i c r o a n a l y t i c a l

Wheatridge,
Terrace,

Colorado

Hackensak,

or G e l l e r

adaptable

for

as

(25)

formed at
5,

and 8 ).

120,

lp73 B l a n c h a r d

(e.g.

fitted

d e t e r m i n a t i o n of

a s c a t a l y s t was us e d f o r

oxidations.-

were

three

O.

Box 1 2 5 ,

"double bonds").

w i t h h o i 1o w - s t o p p e r e d v e s s e l s

d e s c r i b e d by M i l t o n and W a t e r s

Periodate
b y Dy e r

Laboratories,

following hydrogenation reactions

duced p l a t i n u m o x ide
Procedure

P.

New J e r s e y .

M i c r o h y d ro g e n a t ion
A Warburg a p p a r a t u s

Laboratories,

used.

Procedure

The p e r i o d a t e

and w i t h

(62) was f o l l o w e d .

a nd r e a g e n t s

as given

o x i d a t i o n s were p e r ­

hydrogen

ion c o n c e n t r a t i o n s

A l i q u o t s were

titrated

after

as d e s c r i b e d

re­

hydrogenation.

different

and 600 m i n u t e s

is

3,

6,

12,

in the p r o c e d u r e .

(pH 2,

2 If, lf8 ,

2U
Test

for

a c e t o n e . - The

iodoform t e s t

from the p r o c e d u re

d e s c r i b e d by C h e r o n i s

Reagents

t h o s e m e n t i o n e d by t he

used were

the procedure

was

scaled

down s u c h t h a t

c o mp o u n d a n d r e a g e n t s w e r e

was an a d a p t a t i o n

and E n t r i k i n
t wo a u t h o r s ;

only a tenth

1o s i s b e c a u s e

of

its

long

an e x t e n d e d b a c t e r i a l
fast-growing

by t h e

dicated with

activity

s p e c t r u m was

substance
substances

a sufficient

b y D r . R.

ities

are

expressed

per m i l l i l i t e r
the

degree

as the

bacterial

As t h e

this

in o r d e r
to

reason

to find

some e x t e n t

I t was f o u n d

in­

that

the
in­

The a s s a y s w e r e d e s i g n e d
at

of H e a l t h .

be f o u n d

of c u l t u r e

wa y *

least

For

tubercu-

o f c e r t a i n t y by t e s t i n g

Y. G o t t s h a l l

the D i v i s i o n of L a b o r a ­
A d i s c u s s i o n of

in t h e Appendix

least

the

I.

inhibits

activity

highest

growth.

is a ls o

d i l u t i o n which

the

The a c t i v ­

n u mb e r o f m i c r o g r a m s

b ro th which

introductory experiments

in a r e l a t i v e

a nd D i -

o f _5. p s e u d o c a p s i c u m c o u l d be

Michigan Department

methods of a s s a y w i l l

at

studied

in q u e s t i o n .

a g a i n s t D ip 1o c o c c u s p n e umon i a e .
and d i r e c t e d

a g a i n s t M.

incubation period.

organisms which are

ant i tuber cul ar

tories,

followed by

I s o l a t i o n work c o u l d have b e e n hamper ed

b y S_. p s e u d o c a p s i c u m 1 s s p e c i f i c

hibited

of both

assay-"- a g a i n s t M y c o b a c t e r i um t u b e r c u l o s i s

p l o c o c c u s p n e umon i a e .

but

used.

B i o a s s a y . - Th e p u r i f i c a t i o n p r o c e d u r e s w e r e
biological

(20).

( ^ )
In

expressed
inhibits

growth.

"' The a u t h o r w o u l d l i k e t o t h a n k M r s . S h i r l e y G e i s f o r
c o n d u c t i n g t h e b i o a s s a y s on s a m p l e s f r o m t h i s p r o j e c t .

25
C. Experimental

Procedures

ISOLATION WORK
1.
Lucas
of

Prelim inary Experiments

and c o - w o r k e r s

the m acerated r o o ts

antibacterial

(32)

found

that

o f S o l a n u m p s e u d o c a p s i cum c o n t a i n e d

principle.

Therefore,

the

first

were m o s t l y c o n c e rn e d w i t h f r a c t i o n a t i o n
into

different

active

w a t e r - immiscibie
to the water

fractions.

s o l v e n t s or

extract,

for

these

introductory experiments

more,

the

released

soluble

mo r e

of

s t u d y was u n d e r t a k e n

to

and t e s t i n g

the

extract

the

active

in e th a n o l

than

roots

out.

During

principle

wa s f o u n d

in w a t e r .

Further­

re -e x tra c te d with ethanol

antibacterial

investigate

solvents

of the p r e c i p i ­

a c t i v i t y wa s c a r r i e d

once w a t e r - e x t r a c t e d
still

a water

By e x t r a c t i o n w i t h v a r i o u s

collection

antibacterial

of

the

experiments

a d d i t i o n of d i f f e r e n t

tates

c o n s i d e r a b l y mo r e

an a q u e o u s e x t r a c t

principle.

the

Thus a

efficiency

of v a r i o u s

soIvent s .
a.
dried
priate

Extraction

ag en ts. - Fifteen-gram portions

r o o t powder were e x t r a c t e d w i t h
solvent

for

l^O m i n u t e s

in a ^ S o x h l e t

obtained

e x t r a c t s were f i l t e r e d ,

pressure

in a f l a s h

of r o o t weight

to

evaporator

solvent

150 m l .

to

vol ume o f

1:1.

r o o t p o w d e r w a s r e - e x t r a c t e d , now w i t h
solvent.

The e x t r a c t s w e r e f i l t e r e d

to

under

give

the

appro­

apparatus.

concentrated
15 m l .

of the

of

The

diminished

a proportion

The o n c e - e x t r a c t e d

15 0 m l .

of

a different

and c o n c e n t r a t e d

as

26

before

to

terial

activity.

made w i t h

l£ ml.

All

c o n c e n t r a t e s were

The a n t i b a c t e r i a l

different

solvents

c a n be

tested

activity

for

of

antibac­

the

seen from the

extracts

data

given

i n Tab i e I .
TABLE 1
RESULTS OF VARIOUS EXTRACTION SOLVENTS WITH
ROOTS OF S. PSEUDOCAPSICUM
15 g .
Root
Extraction
Sample
Order
No.

I n h i b i t Growth o f :
D.pneumoniae M . t u b e r c u l o s I s
i n a D i l u t i o n Tn a U 1 i u t i o n
of
of

Extraction
S o l v e n t - ”-

1 , e x t r a c t ion

ab so 1 . e t h a n o 1

2 . e x t r a c t ion

water

1 . e x t r a c t ion

80% ethanol

2 . e x t r a c t ion

1 : 61+0

1 : 1021+

1.

i n a c t ive

i n a c t i ve

> 1 : 128

>1 : 1 2 8 0

water

i n a c t i ve

i n a c t i ve

1 . e x t r a c t ion

water

1:61+

1 :80

2 . e x t r a c t ion

Q0% e t h a n o l

1 : 61+

1:80

2.

3.

A ll s o l u t i o n s have a r e l a t i v e
t o 1 ml. o f s o l v e n t .
b.

Possibility

of

concentration

an a n t i b a c t e r i a l

o f 6 N ammoni um h y d r o x i d e

or

straw yellow color

ethanolic

of

the

acid

to

the

1 g.

root

extract

appeared.

same e x t r a c t

of

roots

a lk a lo id . - Addition

6 N sodium h y d ro x i d e

y e l l o w and a f l u f f y p r e c i p i t a t e
hydrochloric

of

changed the
to

a deep

A d d i t i o n of 6 N

did not change

its

appear a n c e .
Since
tained

all

Solanum s p e c i e s

alkaloids,

investigated

i t was t h o u g h t

that

so f a r

have c o n ­

the p r e c i p i t a t e

appearing

27
after

addition

of

a mmoni um h y d r o x i d e

c o u l d be

A drop of Mayerr s r e a g e n t ,

a common a l k a l o i d

added

extract

to

ethanol

1 ml.

of

(Table

the

root

1 ) gave

of

of

changed to

the

solution

a c tio n between
in t h e

an a l k a l o i d .

iodine

(refer

a blue.

This

the

color

T h i s ma y be due t o

reagent)

and

cent

indicated

some t i m e

a re­

starch present

of the water

extract

p r e p a r e d by e x t r a c t ­

r o o t s p r e v i o u s l y e x t r a c t e d w i t h 80 p e r
w ith Table

Mayer1 s r e a g e n t .

I)

gave o n l y

This

c.

a very

indicates

most p a r t were e x t r a c t e d

the

Fractionation

that

first

using

schematically represented

as fo llo w s :

I

of r o o t s
non-ionic

the

cloudiness with

alkaloids

in a batch p ro cess

)

r egener at ion.
w i t h 2N NaOH

—:-----------------1.

the

ion-exchange

a nd may be

* c a t i o n s as
salts

^ , ♦ ,
Dowex 50( H )
Na-salts

---------- 1

Sc he me o f

for

substances)

Dowex 50 (H*") ____________r e g e n e r a t i o n w i t h
(cations)
2 N HC1
▼
Dowex l ( OH
(anions)

ethanol

i o n - e x c h a n g e . - The

out

Ethanolic e x tra ct
(anions, cations,

slight

cent

time by e t h a n o l .

f r a c t i o n a t i o n was c a r r i e d

Figure

wh e n

solution.

One m i l l i l i t e r
ing t h e

After

(from the

reagent,

p r e p a r e d w i t h 80 p e r

a white p r e c i p i t a t e .

the p o s s i b i l i t y

an a l k a l o i d .

ion-exchange f r a c t i o n a t i o n

free

acids

v n o n - i o n ic
substances.
of

root

extract.

28
Dowex 50 w a s

in the

free

T w e n t y g r a ms was p r e p a r e d
time w i t h

100 m i .

with water

until

0.05 N s i l v e r
ium b a s e
each

time with

with water

by w a s h i n g

the washings

did not

Dowex 1 wa s

T w e n t y grams of
100 m l .

until

resin

acid,

to

describes

(RSO^OH) .

twice,

each

a nd t h e n w a s h i n g

give p r e c i p i t a t e

with

i n t h e q u a t e r n a r y ammon­
r e s i n wa s w a s h e d t w i c e ,

of N sodium h y d r o x i d e ,

neutral

The f o l l o w i n g

the

acid form

the

of 2 N h y d ro ch lo ric

nitrate.

form.

sulfonic

and f i n a l l y

litmus paper.
stepwise

the procedure

of

sepa-

r a t ion:
Step
300 m l .

1.

o f 80 p e r

three

hours.

rator

to

50 r e s i n

per

and gave

1.85.

in a S o x h l e t

was c o n c e n t r a t e d
a solution

t h e pH wa s

The

with

apparatus for
in a f l a s h

evapo­

o f pH

extract

wa s m i x e d w i t h

t h e Dowex

on a Be c k ma n p H - m e t e r .

1 . 9 a nd b o t h a f t e r

three

r e s i n w a s now w a s h e d w i t h 50 m l .

a nd f i v e
o f 80

ethanol.

Step

3.

- The e f f l u e n t

t h e Dowex 1 r e s i n .

wa s w a s h e d w i t h 5 0 m l ,

of 80 p e r

s u b s t a n c e s were

evaporator.

cent

added to

one m i n u t e i | . 0 ,

6 .5 and 9 . 5 , r e s p e c t i v e l y .

S t e p 1+. - The e f f l u e n t
non-ionic

p l u s wash l i q u i d s wer e

Th e pH wa s a f t e r

a nd f i v e m i n u t e s

a flash

r o o t m e a l wa s e x t r a c t e d

a n d t h e pH c h a n g e f o l l o w e d

cent

three

ethanol

- The e t h a n o l i c

one m i n u t e

minutes

cent

The e x t r a c t

150 m l .

Step 2.

After

- T h i r t y grams o f

and a f t e r

The r e s i n

ethanol.

p lu s washings which c o n ta in

evaporated

a t 50°C.

The r e s i d u e w a s d i s s o l v e d

the

to dryness

in

i n 30 m l .

of

29
80 p e r

cent

ethanol

Step 5.
time w ith

and t e s t e d

100 m l .

tralization,

of

2 N hydrochloric

e t h a n o 1 was added

o f 80 p e r

contain

the

bacterial

cent

dryness.

to produce
This

each

greenish
After

a final

neu­

volume of

s o l u t i o n presumed to

o r g a n i c c a t i o n i c m a t e r i a l s was t e s t e d

100 m l .

of organic

o f N sodium h y d r o x i d e .

acids present

free

a c i d s by adding

free

acid

form.
to

passed over
the

ethanol

to near

twice,

a nd t h e

6 . - The Dowex 1 r e s i n wa s g e n e r a t e d

time w ith

ness;

ethanol.

acid

activity.

for

anti­

activity.

Step

addition

antibacterial

- The Dowex 5 0 r e s i n wa s f e j e n e r a t e d

c o l o r e d e f f l u e n t was e v a p o r a t e d

30 m l .

for

the

in t h e

t h e Dowex 5 0 r e s i n ,
the

resin.

and

then te s te d

The a c t i v i t i e s

of

sodium s a l t s

t o a Dowex 50 r e s i n

of a green ish color
were c o l o r l e s s

after

The e f f l u e n t w a s e v a p o r a t e d

r e s i d u e was d i s s o l v e d
for

each

e f f l u e n t s were c o n v e r t e d

effluents

The e f f l u e n t s ,

The

twice,

i n 30 m i .

o f 80 p e r

antibacterial

activity.

the f r a c tio n s

are

given

to

on t h e

before
having
to dry­
cent

in T a b l e

II.

TABLE I I
ANTIBACTERIAL ACTIVI TY OF FRACTIONS OBTAINED BY ION-EXCHANGE
Fraction-"Inorganic cations
n i t r o g e n o u s compounds
Free
Non

acids
ionic

Inh ib i t Growth of :
D. p n e u m o n i a e
M. t u b e r c u l o s i s
in a Di 1ut i on of
in a D i l u t i o n of
1:8

inactive
substances

inactive

A l l f r a c t i o n s had a r e l a t i v e
to 1 m l . of s o l v e n t.

c o n c e n t r a t i o n of

1:5
i n a c t ive
i n a c t ive
1 g.

of r o o t s

30
d.

Extraction

i t was f o u n d
ionic

that

the

substances,

In t h e

p r i n c i p l e s . - As r e p o r t e d

a c t i v i t y was a s s o c i a t e d w i t h

which would

following

by e t h e r

of ba sic

experiment

include

nitrogenous

isolation

the

above

cat­

compounds.

of the b a sic

principles

e x t r a c t i o n was a t t e m p t e d .

Thirty

grams o f

r o o t p o w d e r wa s e x t r a c t e d

a p p a r a t u s w i t h 300 m l .
The e t h a n o l

of 80 p e r

cent

e x t r a c t wa s c o n c e n t r a t e d

ethanol

a n d ma de d i s t i n c t l y

i t was e x t r a c t e d

times with ethyl

w i t h 250 ml.

and t h e

The e t h e r - e x t r a c t e d

next

four

for

in a f l a s h

t o a v o l u m e o f 50 m l .
five

in a S o x h l e t

evaporator

ammoniacal.

ether,

times with

2 hours.

Then

the f i r s t

time

100- m l . p o r t i o n s .

w a t e r p h a s e wa s t e s t e d

for

antibacterial

a c t iv i t y .
The c o m b i n e d e t h e r
sodium s u l f a t e

and e v a p o r a t e d

wa s r e d i s s o l v e d
aliquot

(or

activity.
erator

i n 300 m l .

50 m l . )

wa s d i s s o l v e d

t o r emove

of e th y l

was e v a p o r a t e d

i n 5 nil • o f e t h a n o l

over

wa s p a s s e d
extract.

tered

off

through

the

ether

and a o n e - s i x t h

to dryness.
and t e s t e d

A white p r e c ip ita te
ether

w a s 275 itiq -

To t e s t

the p r e c i p i t a t e

was d i s s o l v e d

The a n t i b a c t e r i a l
ethanolic

extract

This residue

for

antibacterial

gas from a gen­

and dr i ed .
for

of

seen

of

the

o c c u r r e d w h i c h wa s f i l ­
The w e i g h t

antibacterial

i n 2 5 ni l .

activity
c a n be

The r e s i d u e

remaining f i v e - s i x t h

and w a s h e d w i t h

precipitate

anhydrous

solvent.

A s t re a m of d r y hydrogen c h l o r i d e

ether

the

e x t r a c t s were d r i e d

of

the

activity

of e t h a n o l .

the v a rio u s
in T a b l e

III.

fractions

of

31
TABLE I I I
ANTIBACTERIAL ACTI VI TY OF FRACT IONS OBTAINED
FROM A ROOT ETHANOL IC EXTRACT
I n h i b i t Gr o w t h o f :
D . p n e umon i ae
M. t u b e r c u l o s i s
in a D i l u t i o n o f :
in a D i l u t i o n of

a)
Fr a c t i on

Ether

e x t r a c t e d water phase

1:8

1:5

Bas ic

substances

in e t h e r

1 : 6 J4-

1 : 160

hydrochloride

1 :256

1 : 61+0

Cr ude a l k a l o i d

E t h e r , a 1k a l o i d f r e e

a) A l l f r a c t i o n s h a v e a r e l a t i v e
r o o t s to 1 ml. of s o l v e n t .
Fr om T a b l e
the

alkaloid
2.

Isolation

-

only the

reported

roots

a nd P u r i f i c a t i o n

that

which

the

activity

1 g.

activity

of

followed

o f So 1a n o c a p s i n e

in e x t r a c t s

leaves,

in t h e p l a n t .

and a s s o c i a t e s

and
(32)

i t wa s d e c i d e d

grams d r i e d

i t wa s

principle

wa s c l o s e l y r e l a t e d

Since

alkaloid

in th e p l a n t ,

antibacterial

Twenty-five

from r o o t s

In t h e p r e c e d i n g p a r a g r a p h

a n t i b ac t e r i a l

is p re s e n t

also possessed

w e i g h t of

of

o f S_. p s e u d o c a p s i cum s h owe d a c t i v i t y

tuberculosis.

alkaloids

from the

the

I t was o r i g i n a l l y f o u n d by L u c a s

a g a i n s t M.

sine,

c a n be s e e n t h a t

Antibacterial

leaves.

to the

it

concentration

fraction.

a.

that

III

i n a c t ive

inac t ive

to

the

o n l y had b e e n

t e s t whether

the

solanocapisolated
leaves

activity.
and g r o u n d

r o o t meal w e r e e x t r a c t e d

for

leaves
four

a nd t h e

hours,

each

same
in a

32
Spxhlet
ml.

apparatus.

o f 80 p e r

Each 25-g.

cent

ethanol.

sample was e x t r a c t e d w i t h 250

Bo th e x t r a c t s were c o n c e n t r a t e d

in a f l a s h

evaporator

t o 25 nil.

minations,

tested

antibacterial

for

and,

after

dry weight

deter­

activity.

TABLE IV
ANTIBACTERIAL ACTI VI TY IN EXTRACTS FROM ROOTS AND LEAVES

Plant Part

mg. Dry W t . / m l .

Leave s

56

-a- Se e A p p e n d i x f o r

details

Th e a c t i v i t i e s
one p a r t
of th e

of

material

the p l a n t

dissolved

activities
contain

I4I 4.0

s hown

antibacterial

b.

in Table

mained

this
t o be

loid present

But

antibacterial

except

other

Different

alkaloid

of

it

is confirmed

as a source

that

crude

alkaloid

changes,

leaves

too

the

isolation

f r o m r o o t s . - As a r e s u l t
of

had any a n t i b a c t e r i a l

roots.

the

the

principle.

a s c e r t a i n e d whether

fo r minor

that

amounts of b a l l a s t

p s e u d o c a p s i c u m was a t t e m p t e d .

in t h e

indicate

d u r i n g e x t r a c t ion c o u l d have changed

of p r e v i o u s c o n s i d e r a t ions,

whether

of u n i t s .

IV do n o t

than the

principle.

Isolation

o f S.

J+U^

a nd d e f i n i t i o n s

is b e t t e r

reported.

the

31+0

8

108

Root s

roots

I n h i b i t Growth o f ;
D. p n e u m o n i a e
M. t u b e r c u l o s i s

the

Besides

from

determining

activity,

s o l a n o c a p s i n e wa s t h e

The m e t h o d u s e d f o r
that

alkaloid

it

re­

alka­

i s o l a t i o n was,

g i v e n by S c h l i t t l e r

a nd

33
Uehlinger
leaves.

(78)

for

The m e t h o d

the

i s o l a t i o n of

used

is o u tlin e d

so 1a n o c a p s i n e
in F i g u r e

from the

2.

I4.OO g . d r i e d a nd g r o u n d r o o t s
p e r c o l a t e d w i t h 6 1 . of 80 per c en t e t h a n o l

r

Ethanol e x tr a c t
(Te s t n o . 9 5 )

Re s i d u e
(d i scarded)

1) C o n c e n t r a t e d t o 2 5 0 m l .
2) Made a m m o n i a c a l
3) E x t r a c t e d w i t h 5 0 0 m l . a n d
5 t i m e s w i t h 250 ml. p o r t i o n s

Ether

of e t h y l

extract

ether

Aqueous
ammoniacal
1aye r
( T e s t n o . 96)

1) E x t r a c t e d w i t h s m a l l p o r t i o n s o f
ten per cent a cetic acid.
U s e d 1250 ml

Ac i d e x t r a c t

Ether e x tra c t
1a y e r
(Te s t n o . 9 7 )

1) N e a r l y s a t u r a t e d b y a d d i t i o n
o f 370 g . s o d i u m c h l o r i d e
I

10 g.

Figure

crude a l k a l o i d hyd ro ch lo rid e
( Te s t n o . 9 9 )

2.

S c h e me o f

isolation

of

alkaloid

Mother 1 i qu or
(Test n o . 9 8 )

from r o o t s .

3J+
Dried
of 80 p e r
lator

and
cent

ground r o o t s
ethanol

equipped with

prevent

the

i_l_8 h o u r s ,
minute

glass

r o o t powder

from p l u g g in g

a speed of

After

10 d r o p s p e r

to

The c o n c e n t r a t e wa s made d i s t i n c t l y

ammoniacal

and

once w i t h
of

%00 m l .
the

separatory funnel
acetic
used.

acid

for

until

of

to

tested

antibacterial

i n a n a mo u n t

(370

the

sufficient

erator

the

alkaloid

crystalline

cent

in a b s o l u t e

acetic

h y d r o c h l o r i d e was

standing

antibacterial

activity.

acid

p h a s e wa s

ethanol

a nd

10.0

acid

overnight

solution

the

alkaloid

in the

refrig­

h y d r o c h l o r i d e wa s c o l l e c t e d

g.

sodium c h l o r i d e .

a nd t h e

acetic

ether

to p r e c i p i t a t e

a t room t e m p e r a t u r e .

hydrochloride

10 p e r

activity.

After

of

10 p e r

in a 2 - l i t e r

amounts of

extracted

dissolved

hydrochloride.

a mo u n t

small

wa s a d d e d t o t h e

g.)

and d r i e d

of

acid

as the

on a f i l t e r

with

ammoni a­

activity.

e x t r a c t ' s were p l a c e d

evaporated

Sodium c h l o r i d e

a nd 5 t i m e s w i t h

The r e m a i n i n g

antibacterial

1250 ml.

dryness,

ether

solvent.

a nd e x t r a c t e d

An a l i q u o t

for

of ethyl

same

The c o m b i n e d e t h e r

a large

stop cock.

to

( 5 * 6 1 . ) wa s o b t a i n e d .

was c o n c e n t r a t e d

p h a s e wa s t e s t e d

alkaloid

at

of c o t t o n

evaporator

25 0 m l • p o r t i o n s

wa s

the

glass perco­

in a f l a s h

extracted

cent

in a c o n i c a l

s t o p c o c k a nd a p l u g

and a g o l d e n br own e x t r a c t

250 m l .

w e r e m a c e r a t e d w i t h 6 1.

J4.8 h o u r s

p e r c o l a t i o n wa s p e r f o r m e d

Th e e x t r a c t

cal

for

(I 4.OO g . )

a c i d i c mother

The y i e l d

of

crude

and f o u n d c o n t a m i n a t e d w i t h
Both the

crude

l i q u o r were

alkaloid

tested

for

35

Two g r a m s o f c r u d e
by h e a t i n g
tinctly

of

over

to give

of w a te r.

h y d r o c h l o r i d e was d i s s o l v e d
The w a t e r

ammoni acal was e x t r a c t e d

portions
dried

i n 20 m l .

alkaloid

ethyl

anhydrous

2 6 0 mg.

activity

ether.

sodium s u l f a t e

base.

and t h e

ether

base

wa s d e t e r m i n e d .

The

antibacterial

fractions

seen

100-ml.

e x t r a c t s were

and e v a p o r a t e d

alkaloid

the

c a n be

times with

The c o m b i n e d e t h e r

of crude

of both

three

s o l u t i o n made d i s ­

to dryness

The a n t i b a c t e r i a l
e x tr a c te d water phase

activity

of t h e v a r i o u s

i n T a b l e V.
TABLE V

ANTIBACTERIAL ACTI VI TY DURING ROOT ALKALOID ISOLATION
8/ml,
Test
no.

Fraction

95

Ethanol

96

Ammoniacal

97

Ether

98

Mother

99

Crude

mg. D r y
Wt./ml.

extract
layer

extract

layer

1 iquor
a l k a l o i d HC1

100

A l k a l o id b a s e

101

Aqueous phase from
e x t r . of a lk a lo i d
base

c * Isolation
b a tc h e s of

of

I n h i b i t Growth o f :

D.pneumoniae

M.tuberculosis
On
On B l o o d
Agar
Agar

138

30

5k

188

>100

>100

>100

i n a c t i ve

inac t

k- .6

72

5 5 .>5

>100

>100

>100

2 .,0

16

25

25

1 .,0

16

12

12

>100

85

so 1 a n o c a p s i n e

so 1 a n o c a p s i n e w e r e

i n a c t ive

inac t

from l e a v e s . - S e v e ra l

isolated

during

this

in v e s tig a t ion.

36
Th e p r o c e d u r e
Schlittler

and U e h l i n g e r

Dried
of 80 p e r
The

w i t h m i n o r m o d i f i c a t i o n s wa s t h a t

and g r ou n d
cent

leaf material

(78)*

leaves

ethanol

(500 g . )

was f i l t e r e d

for

combined,

deep g re e n f i l t r a t e s

chlorophyll
coating.

1+8 h o u r s w i t h

to

1250 m l .

separated

in th e

refrigerator

ether

of

concentrate.

times with

the

three

extractions

After

em ulsification.

extract

were p o s s i b l y

amounts of

10 p e r

e x t r a c t s wa s a d d e d

iness p e rs is te d
frigerator.

and t h e

After

in a f l a s h
amounts of

as a v i s c o u s
acetic

wa s r e m o v e d b y f i l t r a t i o n .
into

fractions

In a 2-1 i t e r

However,
to

o f 50 0 ml.

separatory

1 1 . of

The e t h e r

acid.

the

last

separate

the

t wo p h a s e s

lipids present

t wo

in t h e

ammoni um h y d r o x i d e .

To t h e

sodium c h l o r i d e

(330 g.)

the p r e c i p i t a t e

small

combined a c e t i c

s o l u t i o n wa s t h e n p l a c e d

10 h o u r s

phase

during

e x t r a c t s were e x t r a c t e d w i t h
acetic

ethyl

during

s a p o n i f i e d by t h e

cent

The

standing overnight

once w i t h

Some o f t h e

The c o m b i n e d e t h e r

ethanol.

a q u e o u s a mmoni a l a y e r

i t was v e r y d i f f i c u l t

and a g a i n

glacial

500- m l . p o r t i o n s .

extractions.

due t o

flask

9 1*

f o r 1+8 h o u r s .

large

200 ml.

ammoniacal.

r e a d i l y from the

first

the

was t h e n d i v i d e d

separated

acid

side

e a c h f r a c t i o n wa s e x t r a c t e d
and f i v e

cent

were c o n c e n t r a t e d

chlorophyll

a nd e a c h made d i s t i n c t l y
funnel

o f 80 p e r

the chloro p h y ll

The c o n c e n t r a t e

jar

on a B u c h n e r f u n n e l

9 1.

on t h e

the

glass

During c o n c e n tra tio n

To p r e c i p i t a t e

a c i d was a d d e d t o

were m a c e r a t e d w i t h

in a c y l i n d r i c a l

extracted

evaporator

f o l l o w e d by

until
in the

cloud­
re­

wa s c o l l e c t e d

on

37
a filter

and d r i e d

gave o n l y an
The y i e l d

of crude

weight of

the

Several

a 1 .6 per

h y d r o c h l o r i d e was 8 . 0

cent yield

isolations

gave y i e l d s

alkaloid

a large

of

using

1.26 per

hydrochloride

the procedure

cent

and

scale

1 . of w a te r

and t h e n e x t r a c t e d

alcohol

of e th a n o l

for

batch

wa s r e m o v e d b y d i s t i l l a t i o n

wa s f u r t h e r

rator.

The c o n c e n t r a t e wa s d i v i d e d

concentrated

up a s d e s c r i b e d

loid hydrochloride
per

cent yield

and U e h l i n g e r

ether

based

extraction

very

slight

obtained
gave

g.

on d r y w e i g h t
of

(78) noticed

layer

Mayer’ s t e s t .

leaves.

the

and t h e

of

remaining
evapo­

500-ml. fra c tio n s
of crude

corresponding

to

alka­

a 0.11

of r o o t s .
extraction. - Schlittler
concentrated

f r o m an e x t r a c t

particular

apparatus

in a f l a s h

The y i e l d

a highly positive

When t h i s

in the

The m a i n p o r t i o n

into

alkaloid
that

r o o t m e a l wa s a l l o w e d

to 2 1 .

above.

wa s o n l y 2 . 2

d * The c o m p l e t e n e s s

ammoniacal

of

seven ho u rs.

extract

and w o r k e d

cent

i s o l a t i o n wa s a t t e m p t e d b y m a k i n g u s e o f

to absorb

the

of

des­

e x t r a c t o r . - To s i m p l i f y t h e c u mb e r s o m e p r o ­

A 2 kg.

12 1.

1 . 51+ p e r

b a s e d on d r y w e i g h t

extractor.

with

g.,

b a s e d on t h e d r y

a Barnstead
2

liquor

leaves.

Barnstead
cedure

to

The m o t h e r

c l o u d i n e s s w ith Mayer’ s re a g e n t .

s o 1a n o c a p s i n e

additional

above

of crude

room t e m p e r a t u r e .

insignificant

which c o r r e s p o n d s

cribed

at

of

a nd a q u e o u s

leaves

alkaloid

after

test

with

s t e p was c h e c k e d o n l y a

c l o u d i n e s s w i t h M a y e r ’ s r e a g e n t was o b s e r v e d .

Nevertheless,

i t was d e c i d e d

to r e - e x t r a c t

the

ether

extracted

38
ammoniacal

concentrate

determine

i f mo r e

For

this

by Mo rt o n

alkaloid

purpose

it

is

tion

a liquid with

a 1 i q u i d - 1 iquid

a heavier
formed

the

increased

the mother

in a s e p a r a t o r y f u n n e l .
■l i quor w e r e e x t r a c t e d
c h l o r o f o r m and t h e

five

liquid;

but

for

in t h i s

extrac­

case

than organophilic

Since

t h e vol ume of

i t wa s n e c e s s a r y t o

Purification

part

of

gram o f

of

the

c a p s i n e was g r e a t e r
fraction,

of t h e mother

t i m e s w i t h 2 5 0 - ml . p o r t i o n s

gas.

used

the

to give

in t h i s

of

saturated

alkaloid

a yield

of

1.1

e x p e r i m e n t wa s o r i g ­

of r o o t meal,

hence

the y i e l d

in

on d r y w e i g h t .
so I a n o c a p s i n e . -

antibacterial

than the

the

The p r e c i p i t a t e d

and d r i e d

f r o m 500 g.

determine whether

wa s e x t r a c t e d w i t h c h l o r o f o r m

c ombi ned c h l o r o f o r m e x t r a c t s were

c e n t wa s 0 . 2 2 b a s e d

alkaloid

( 2 : 1 ).

of e x t r a c t o r

Two 7 5 0 - m l . p o r t i o n s

concentrate

inally obtained

One

type

concentrate

an a z e o t r o p ic m i x t u r e .

liquor

was f i l t e r e d

The a m m o n i a c a l

e.

as d e s c r i b e d

of ammoniacal

so much t h a t

dry hydrogen c h loride

hydrochloride

per

to

extraction.

Instead

with

extractor

of c h lo r o f o r m - e t h a n o l

wa s mo r e h y d r o p h i l i c

the e x t r a c t a n t
stop

1250 m l .

a very convenient

c h l o r o f o r m a nd e t h a n o l

mixture

c o u l d be r e m o v e d .

11^.00 m l .

Usually

ethanol

a chioroforin-ethanol

( 614.) w a s u s e d a n d

wa s e x t r a c t e d w i t h

of

with

crude,

activity

activity

preceding

In an a t t e m p t

of

the

of pu re
crude

crude m a te ria l

i n 70 m l .

solano-

alkaloid

wa s p u r i f i e d .

greenish preparation

h y d r o c h l o r i d e was d i s s o l v e d

to

of

leaf

of 80 p e r

g.

39
cent

ethanol,

heated

to

60-70°C.

The f i l t e r e d

s o l u t i o n was t e s t e d

to determine

if

ter ial

the

active

and d e c o l o r i z e d w i t h N o r i t .
for

antibacterial

c a r b o n had a d s o r b e d

any a n t i b a c -

mater i a l .

The e t h a n o l i c

s o l u t i o n wa s e v a p o r a t e d

pended

i n 3 N ammoni um h y d r o x i d e

funnel

seven tim e s w ith

100-ml.

remaining

a mmo n i a l a y e r

activity,

and t h e

anhydrous

s od i um s u l f a t e

of c o l o r l e s s

wa s

and e x t r a c t e d
portions

saved

combined e t h e r

alkaloid

to dryness,

a nd t e s t e d

for

ocapsine

of m e l t i n g

point

a c o mp o u n d m e l t i n g

to

the m a t e r i a l s

for

212-211| °C.

The y i e l d

Recrystalliza­

crystalline

solan-

A second r e c r y s t a l l i z a t i o n

2 1 5 > - 2 l 6 ° C.

antibacterial

The

antibacterial

dryness.

o b t a i n e d wa s l\.^0 mg.

at

ether.

f r a c t i o n s were d r i e d w i t h

and e v a p o r a t e d

base

sus­

in a s e p a r a t o r y

of e t h y l

t i o n f r o m e t h a n o 1- w a t e r ( 1 : I ) gave c o l o r l e s s ,

gave

activity

The r e s u l t s

activity

c a n be

of

seen

testing
in T a b l e

VI .
T a b l e VI
alkaloid
its

s ho ws t h a t

increased

activity

the

a n t 1t u b e r c u l a r

four-fold

against

D, p n e union i ae d e c r e a s e d

T h i s ma y s u g g e s t

that

besides

so 1 a n o c a p s i n e wa s p r e s e n t

another

w h i c h wa s d i f f i c u l t

to

sine

c o u l d be o b t a i n e d

while

one a n d o n e - h a l f

antibacterial

of d i f f e r e n t

resinous material

crystallize.

a nd c a u t i o u s p r e c i p i t a t i o n

the

c o mp o u n d

in th e p l a n t .

Often during r e c r y s t a l l i z a t i o n
an a mo r p h o u s ,

of

during the p u r i f i c a t i o n ,

times.

so 1 a n o c a p s i n e

activity

b a t c h e s of

wa s e n c o u n t e r e d ,

But by r e p e a t e d

solution

from e t h a n o l - w a t e r ( 1 : 1 ) solanocap-

in c r y s t a l l i n e

form.

l+o
TABLE VI
ANTIBACTERIAL ACTI VI TY DURING SOLANOCAPSINE PURI FI CATION
^ml .
Fraction

mg. D r y
Wt./ml.

Cr u d e a l k a l o id
h y d r o c h 1o r i d e

I n h i b i t Growth o f :

D . p n e umon i ae

M.tuberculosis
On
On B l o o d
Agar
Agar

1

20

13

13

Decolorized alkaloid
h y d r o c h l o r ide

lit

l it

kk

22

Ether e x tra cted
a mmo n i a l a y e r

20

78

>100

A lk a lo id base
( s o l a n o c ap s i n e )

2

8

25

25

Solanocaps in e ,
r e c r y s t a l 1 i zed
f o ur t ime s

o.kS

30

3

3

f.

C o l u mn c h r o m a t o g r a p h y . -

task

to p u r i f y crude

were

difficult

carbon.

to

so 1 a n o c a p s i n e .

remove,

Only through repeated

f r o m e t h a n o 1 - w a t e r ( 1 : 1 ) wa s
pound;
decided

it possible

the

y i e l d wa s e x t r e m e l y

to

try

separation

t i o n on a l u m i n a ,

acid

r o o t s wa s u s e d f o r
preparation

a n d Dowex 5 0 .

alumina,

alumina,

from the

the

small.

impurities
activated

so 1 a n o c a p s i n e
a p u r e com­

Therefore,

i t wa s

f r a c t i o n by ad so rp ­

o r Dowex 50 •
a crude

r o o t s wa s u s e d f o r

of

to o b tain

alkaloid

whereas

a very d i f f i c u l t

treatment with

crystallization

but

of

I t wa s f o u n d
The c o l o r e d

even a f t e r

inac t ive

An e x t r a c t

of

the

so 1 a n o c a p s i n e

trials

on a c i d

alumina

C o l u m n c h r o m a t o g r a p h y on a l u m i n a . - P r e p a r a t i o n o f e x ­
A 175 9* p o r t i o n

tract:
1500 m l .

tractor

of 80 p e r

cent

(described

wa s c o n t i n u e d f o r

ness of

extraction.

r o o t powder

liquid

over

vol ume o f

the

ethanol

seven hours

However,

the

The

that

i t was

root

sample.

1270 ml .

to

and f i l t e r e d ,

^/ml . against

column

in small

wa s t a p p e d
all

the

the

of

effective­

so r a p i d l y

to m a in ta in

through

a head of

o b t a i n e d had a

activity

of

)f/m l .

3 30

e x t r a c t was c o n c e n t r a t e d

antibacterial

a c t i v i t y b e c a me

same o r g a n i s m .

p a c k e d " w i t h 2 0 0 g.
following

about

The e x t r a c t

When t h e

of e x ­

The e x t r a c t i o n

flowed

impossible

type

co mp lete removal

existed

solvent

the

C o l u mn p r e p a r a t i o n :

in t he

secure

and an a n t i b a c t e r i a l

tuberculos i s .

125

in a p e r c o l a t o r

to

some d o u b t

a g a i n s t M.
1 2 5 ni l .

r o o t powder was e x t r a c t e d w i t h

in A p p a r a t u s S e c t i o n ) .

alkaloids.
the

of

The 50 x 2 . 1

of Alcoa alumina,

fashion.
portions

gently with

The

cm.

grade F-20,

8 0 me s h ,

a l u m i n a was p o u r e d

into the

and a f t e r

each a d d it i o n ,

a rubber m allet.

alumina a f i l t e r

paper

c o l u m n wa s " d r y -

After

d i s k wa s p l a c e d

t h e column

a d d i t i o n of
on t o p o f

the

ad s o r b e n t .
Introduction
ethanol

wa s a p p l i e d

wa s d i s c a r d e d .

into

A b o u t I4.OO m l .

t h e c o l u mn and t h e

When o n l y a 2 mm.

above

the

alumina,

After

the

root

that

of e x t r a c t :

the

extract

o n l y a 2 mm.

root

extract

had p a s s e d

liquid

height

height

of

of a b s o l u t e
first

effluent

solvent

remained

wa s a d d e d t o t h e

so f a r

down

into

rem ained above th e

column.

the

alumina

alumina,

the

c o l u m n was e l u t e d w i t h

20 m l .

were

absolute

collected

ethanol

e v a p o ra tio n of
Table VII
compounds

absolute

until

a nd F i g u r e

(=alkaloids)

wa s a c c o m p l i s h e d .
11 w e r e

in f r a c t i o n

all

It

colored p a rt

2 s h o ws t h a t

followed

3 parts.

Part

and p a r t s

2 and

did.

while

of b a s i c

the

activity.

from a l k a l o i d s

here

that

highest

rest

for

tested

for

five

the

fractions

intensity

in u l t r a v i o l e t

colored

The u p p e r

light,

hours with
to

activity.

the

dryness

l ower

colorless

pneumoniae; but
upper

of dry m a t e r i a l ,

one-fourth

which

o f 80 p e r
in a f l a s h

cent

None o f

into

in t h e y e l l o w zone

300 m l .

of 80 p e r

whereas

a nd d i v i d e d

E a c h s e c t i o n wa s e x t r a c t e d

i n 20 m l .

a g a i n s t D.

in t h e

a l u m i n a wa s o f

wa s c o l o r l e s s .

of t h e m a t e r i a l

evaporated

antibacterial

s h owe d a c t i v i t y

of

The a l u m i n a wa s e x t r u d e d

The e x t r a c t ,

1 mg.

the

segments.

wa s d i s s o l v e d

tained

impurities

3 we r e o b t a i n e d by d i v i d i n g

apparatus

origin

a mo u n t

antibacterial

one-fourth

1 consisted

rator,

its

of

the

s h o u l d be r e p o r t e d

The u p p e r

into 2 equal

ethanol.

the

g a v e no f l u o r e s c e n c e

lower p a r t

Soxhlet

s howed a f t e r

1+.

a deep y e l l o w c o l o r ,

part

Elution with

no r e s i d u e

yellow -colored with

Extrusion:

the

of

solvent.

A p p a r e n t l y n o t much s e p a r a t i o n

1 to

Fractions

in t a r e d 5 0 - m l . b e a k e r s .

wa s c o n t i n u e d

the

ethanol.

ethanol

the
the
of

in a
cent
evapo­

a nd

3 fractions
fra c tio n with

the

column c o n ­

s h o we d a n a n t i b a c t e r i a l

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Ion-exchange

c h r o m a t o g r a p h y . - C o l u mn p r e p a r a t i o n :

The

Dowex SO r e s i n w a s p r e p a r e d b y w a s h i n g

the

a lte rn a te ly with 2 N hydrochloric

and 2 N s o d i u m h y d r o x ­

ide.

After

each

addition

washed w i t h w a t e r

until

of e i t h e r

neutral.

2 N hydrochloric

acid,

first

and t h e n w i t h

with water

now f i l l e d
39 x 2 . 1

into

cm.

the

the

decreased
paper

to

ethanol

was a l l o w e d

the f i n a l

g.

the

of c r u d e

of crude

so 1a n o c a p s i n e

t h e col umn and a l l o w e d

and 5 m l . o f

until

o n l y a 2 mm. h e i g h t

small

volume o f

60 p e r

a nd a g a i n a l l o w e d

were

remained

the

the

ethanol.

resin

resin

The r e s i n ,

dimensions
During

in t h e

settled,

through

neutral

135 m l .

the

r e s i n wa s

washing with

had the

the

column

a filter

and 60 p e r

cent

t h e column t o

above

the

aid

remained

above

ethanol

i n 100 m l .
a c i d wa s

resin.

to flow through,

of

i n 60 p e r

introduced

resin.

resin
Then a

the

developing

and f r a c t i o n s

cent

to

cent

column

collected,

of

solvent

ethanol

of

of

60 p e r

o n l y a 2 mm. h e i g h t

Now t h e

When 1 3 0 0 m l . w e r e
10 p e r

the

was a p p l i e d

down u n t i l

allowed

ethanol.

acetic

A solution

t o f l o w down t h r o u g h t h e

ammoni um h y d r o x i d e

collected.

solution:

from r o o t s

cent

cent

cent

r e s i n had

glacial

cent

to pass

s o l v e n t wa s c h a n g e d t o
per

l e n g t h of

alkaloid

into

added,

a final

a vol ume o f

to p erco late

ethanol

of S per

60 p e r

on t o p o f

cent

liquid

or base

times,

packing.

Introduction
2.2

to

When t h e

d i s k wa s p l a c e d

After

in a s l u r r y ,

corresponding

3l± cm.

acid

several

r e s i n was wa s h e d u n t i l

column

chromatographic process

acid

resin

wa s

100 ml . e l u a t e

the

developing

ammoni um h y d r o x i d e

i n 60

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U7
In a d d i t i o n
density

to dry weight

o f e a c h f r a c t i o n was d e t e r m i n e d

a n d Lomb s p e c t r o p h o t o m e t e r .
intended

to

give

impurities.
spectra

of

an

several

a n d a mo u n t

The

idea about

The w a v e l e n g t h

at

at

the

7 5 0 mji i n a B a u s c h

distribution

in the

7 5 0 mp.

the o p t ic a l

spectrophotometric

o f 7 50 mji wa s

fractions

maxi mum a b s o r p t i o n
sities

determinations,

of c o l o r e d

selected,

since

r e g i o n l; 0 0 - 9 5 0 mp s h o we d

Figure

of dry m a t e r i a l

d a t a were

J_|_ r e c o r d s

in the

color

different

inten­

eluate

f ractions.
Paper

c h r o m a t o g r a p h y was a p p l i e d

to determine
whether

the

o n l y one a l k a l o i d

solanocaps i n e ."
reagent

No s e p a r a t i o n

wa s 0 . 5 7 , t h e v a l u e

of a l k a l o i d

this

in what

of t h i s

of the

fractions

a nd t o d e c i d e
is c a l le d

solvent

and c o l o r

e x p e r i m e n t wa s n e g a ­

alkaloid

i m p u r i t i e s wa s

from a l l

for

fractions

so 1 a n o c a p s i n e .

a c o mp o u n d w i t h R^ = 0 . 14.8 ; b u t

c o mp o u n d w a s e l u t e d

from the paper

"crude

chromatography.

from c o l o r e d

found c h a r a c t e r i s t i c

The ^ w a s h i n g ” c o n t a i n e d

eluted

s e c t i o n on p a p e r

result

The R ^ - v a l . u e

all

so 1 a n o c a p s i n e

wa s p r e s e n t

in th e

The o v e r a l l

accomplished.

of

The c o m p o s i t i o n o f b o t h

is described

Result:
tive.

distribution

to

if

a nd r e - c h r o m a t o g r a p h e d ,

t h e R ^ - v a l u e wa s 0 . 5 7 *
Co l u mn c h r o m a t o g r a p h y on a c i d

a l u m i n a . - The a c i d

u s e d wa s Woe l m' s a n i o n o t r o p i c a l u m i n u m o x i d e ,
1,

and s p e c i f i e d

grams of
Water

the

for

chromatographic

activity

analysis.

1- b u t a n o l

wa s a p p l i e d

to

the

grade

One h u n d r e d

a l u m i n a w a s f,d r y - p a c k e d " a s d e s c r i b e d

saturated

alumina

before.

column b e f o r e

1*8

TABLE V I I I
DATA FOR ELUTION CHROMATOGRAPHY OF CRUDE SOLANOCAPS INE
FROM ROOTS ON ACID ALUMINA

Fr a c t ion
Eluted

Amo u n t
o f Dry
Mater ial
in mg.

J/5nl .
Co l o r
of
F r a c t ion

Inh ib i t s Gr owt h o f :
D. pne umoni ae
M.tube r c u l o s i s

1

1

colorless

i n a c t i ve

i n a c t i ve

2

1

colorless

i n a c t i ve

i n a c t i ve

3

2

colorless

i n a c t i ve

i n a c t i ve

Ip

8

we a k ye 1 1 ow

25

i n a c t ive

5

106

we a k ye 1 1 ow

16

13

6

327

ye 1 1 ow

16

13

7

35k

ye i 1 ow

16

13

8

2kk

we a k y e 1 1 ow

16

13

9

81

we ak ye 1 1 ow

8

6

10

38

we a k ye 1 1 ow

60

k7

11

10

colorless

8

6

12

3

colorless

5

It

13

It

colorless

6

5

Ik

2

colorless

3

2.5

15

3

colorless

5

k

16

2

colorless

3

2.5

17

1

colorless

1.5

1 .25

18

1

colorless

3

2.5

14-9
application

of

so 1 a n o c a p s i n e
tanol

was

height

alkaloid.

from r o o t s

introduced

of

liquid

wa s e l u t e d
of

the

10 m l .

were

for

for

the

the

remained

above t h e

collected,

the

g.

of c ru d e

saturated

alumina the

1- b u t a n o l .

adsorbent

Eluate

fractions

s o l v e n t wa s e v a p o r a t e d

activity.

1- b u ­

All

and t h e

f r a c t i o n s were

Table V I I I

gives

the

data

experiment.

Result:

Some

separation

a l k a l o i d wa s a c c o m p l i s h e d .
s o 1a n o c a p s I n e
i t y ”, but

of c o l o r e d

Fractions

i n d i c a t e d by f a i r l y

i m p u r i t i e s from
consisted

11-18

constant

of pure

antibacterial

activ­

t h e y i e l d wa s e x t r e m e l y s m a l l .

g.

Alkaloid

known t o w h a t
alkaloid

1.2

When o n l y a 2 mm.

was d e t e r m i n e d .

antibacterial

of

of water

column.

saturated

amount o f d r y m a t e r i a l
tested

in 20 ml.

into

with water

A solution

extent

content

in the p l a n t . - Since

so 1a n o c a p s i n e

occurs

d e t e r m i n a t i o n s were c a r r i e d

is an a d a p t a t i o n

of

t h e method

out.

i t was n o t

i n S. p s e u d o c a p s i c u m ,
The p r o c e d u r e

given for Belladonna Leaf

used
in

P h a r m a c o p e i a o f The U n i t e d S t a t e s :
Extraction:
me s h ) w a s p l a c e d
wa s

inserted

Ten grams of d r i e d
in an e x t r a c t i o n

in a S o x h l e t

leaf

thimble,

extractor.

or

r o o t powder

and t h e

( IpO

thimble

The p l a n t m a t e r i a l

wa s

The a c t i v i t y o f t h e f r a c t i o n s c a n e a s i l y v a r y , s i n c e
the d i l u t i o n t e c h n iq u e has e x p e rim e n ta l l i m i t a t i o n s .
Besides,
t h e amount o f d r y m a t e r i a l i s o n l y d e t e r m i n e d t o t h e n e a r e s t
mg.
When t h e a m o u n t o f d r y m a t e r i a l i n a f r a c t i o n i s i n t h e
r a n g e o f 1 mg. a s l i g h t d e v i a t i o n i n t h e w e i g h t d e t e r m i n a t i o n
can e a s i l y change the a c t i v i t y c o n s i d e r a b l y .

50
moistened with

a mixture

ide,

absolute

10 m l .

mixed

of

thoroughly,

ethanol,

and m a c e r a t e d

wa s t h e n c o v e r e d w i t h
material

of 8 ml.

a tuft

was e x t r a c t e d

for

concentrated
a nd 2 0 m l .

for

ammoni um h y d r o x ­

of

ethyl

six hours.

of p u r i f i e d

The t h i m b l e

cotton,

six hours with

ether,

and t h e p l a n t

1^0 m l .

of e t h y l

ether .
Purification:

The a l k a l o i d a l

solution

m a c e r a t i o n was c o n t a m i n a t e d w i t h o t h e r
interfere

with

To e f f e c t

the p u r i f i c a t i o n ,

the

the volum etric

s o l v e n t by e x t r a c t i n g

l u t i o n wa s made
solvent

such as e t h y l

The e t h e r

extract

extractor

ethyl

each time,

separatory funnel.
the e t h e r

layer

of N a c e t i c
bined

acidic

and t h e

free

sive 50-ml.

i t y upon t h e

an a c i d ,

then

a nd e x t r a c t e d

wa s t r a n s f e r r e d
wa s r i n s e d

the

with

aqueous

an

so­

immiscible

a nd t h e s e

to a sep ara to ry funnel;

twice with

about

r i n s i n g s were

filtering

each p o r t io n

s o l u t i o n s were r e n d e r e d
alkaloid
portions

and r e d i s s o l v e d
This

a l k a l o i d s were r emoved f r om

by e x t r a c t i n g w i t h 7 s u c c e s s i v e

acid,

alkaloid.

10 m l .

of

added to the

The a l k a l o i d wa s r e m o v e d c o m p l e t e l y f r o m

t i o n of e x t r a c t i o n ,

acid.

the

the

ether:

the S o x h le t
ether

e x t r a c t i v e s wh i c h woul d

d e t e r m i n a t i o n of

with

ammoniacal

o b t a i n e d by t h i s

wa s
of

1 ml.

addition

ether.

of the

in 0 . 5 ml.

solution

of

distinctly

last

portions

The c o m­

ammoniacal,
with 5 succes­

To d e t e r m i n e

the comple­

e x t r a c t wa s e v a p o r a t e d

approxim ately 0.5 N hydro ch lo ric

should not
of

drawn o f f .

immediately e x tr a c te d

ethyl

10-ml.

show mo r e

t h an a s l i g h t

a drop of Mayer1s r e a g e n t .

turbid­

51
Determination:
ethyl

ether

was d r i e d

and f i l t e r e d .
e t h e r , and

The

solution

by sha king w ith

The f i l t e r

t h e w a s h i n g wa s a d d e d t o t h e

there

by a d d i t i o n

for
of

in a m e a s u r e d
acid
to

to

give

insure

methyl

about
and

red

with

as

years

of e t h y l

solution.

This

d r y n e s s on a s t e a m b a t h

The r e s u l t i n g

1 m l . of

sodium s u l f a t e

10 m l .

ether

in

absolute

and

r e s i d u e wa s s o f t e n e d

ethanol

and d i s s o l v e d

s u f f i c i e n t volume of 0 . 0 2 N h y d r o c h l o r i c

complete

Alkaloid
three

15 m i n u t e s .

an a c i d i c

wa s t i t r a t e d

to

alkaloid

anhydrous

wa s w a s h e d w i t h

s o l u t i o n was t h e n e v a p o r a t e d
heated

of p u r i f i e d

solution.

solution

The m i x t u r e w a s g e n t l y h e a t e d

of the

alkaloid.

0 . 0 2 N sodium h y d r o x i d e

The e x c e s s

acid

in t h e p r e s e n c e

of

indicator.
determinations

c a n be f o u n d

on p l a n t m a t e r i a l

in Table

from the p a s t

IX.

TABLE IX
ALKALOID CONTENT IN S.
Ye a r
of
Harve st

PSEUDOCAPSICUM ON DRY WE IGHT BASIS

N i t r o g e n Level
in S o i l

A 1 k a l o i d P e r c e n t a ge i n :
L e a v e s a nd S t e ms
Root s

1953

normal

1 .53

0.83

1951|

normal

2 . 11|

1 .53

1955

1 ow

2.21

1 .86

19 55

norma1

2.15

2 . 2 l|

These
fairly

determinations

u n i f o r m a mo u n t o f

s h o we d t h a t
alkaloid.

the p l a n t

Seasonal

contains

and o t h e r

a

52
environmental
but

so f a r

t h e y have not

h.
that

isolated

a nd

identical

iso lated by both Barger
and U e h l i n g e r

of ro o t

antibacterial

l e a v e s wa s

(78)

flu ctu atio n s within

a nd

points

and p h y s i c a l

second p a r t

this

table

the

with

with

so 1 a n o c a p s i n e

the

isolated
alkaloid

Details

of t h i s

so 1 a n o c a p s i n e

( 6 ) a nd S c h l i t t i e r

of proce dure

investigation.

are

of both

c a n be

from both

c o n s t a n t s were d e t e r m i n e d

constants

identity

a l k a l o i d . - To c o n f i r m

alkaloid

n u mb e r o f d e r i v a t i v e s p r e p a r e d .
in the

leaf

a n d F r a e n k e 1- C o n r a t

physical

cribed

lim its;

been e x te n s iv e .

Derivatives

the

roots

f a c t o r s ma y c a u s e

reported

the

root

are d e s­

Only m e l t i n g

i n T a b l e X.

a nd

and a

leaf

Fr om

alkaloid

seen.
TABLE X

CHEMICAL CONSTANTS OF ROOT AND LEAF ALKALOIDS
A 1 k a l 0 id s f rom
Leave s
Roots
F ound S c h l i t t l e r B a r g e r
e t al .
e t a l . Found

Determ i n a t i o n
B a s e , m.p .
Dihydrochloride,

215- 6°

>300°

m.p.

>300°

m.p.

Isopropylidene

derivative,

N itro so d e r i v a t i v e , m.p.
T rim ethyl d e r i v a t i v e , m.p.
Nitrogen,
Optical

per

cent

rotation,

3LS"

[oC ]j>

R a s t ' s m olecular weight
de t e r m i n a t i o n
N eutralization

equivalent

0
*—
1
1
0
0

Oxalate,

C\J

19 l p- 6 °

P ic r a te , m.p.

216-7°
>300°

191+°

216- 6°

>300°
198-9°
287-9°

233°
200°

205- 6 °

209°

6.20

6.29

+25°

+ 22j °

J4.82

280°

288-9°

m .p . 232-3°
19lt-6°

i_l_5 6 , Ip81

222°

I9k°
208°

+25.5°
1+60

+23.6°

53
3.

The A n t i b a c t e r i a l

So 1a n o c a p s i n e
sicum gave

isolated

an a n t i b a c t e r i a l

S p e c t r u m o f So 1 a n o c a p s i n e
from the

l e a v e s o f S.

spectrum as

s hown

pssudocap-

in T ab l e X I .

TABLE XI
ACTI VI TY OF CRUDE SOLANOCAPSINE BASE AGAINST
VARIOUS MICROORGANISMS

O r g a n i s m s a n d S t r a i n No.

Aerobacter
Bacillus

aerogenes

Concentration Required
f o r I n h i b i t i o n o f Growth

6l8

500

s u b t i l i s 231

78

Corynebacterium d ip h te ria e
D i p l o c o c c u s p n e u m o n i a e Type
Escherichia coli

PW8
III

621

125
20
50 0

H i s t o p l a s m a c a p s u l a t u m Duke U n i v . , y e a s t p h a s e
H i s t o p l a s m a c a p s u l a t u m Duke U n i v . , m o l d p h a s e

15 - 5
125

K l e b s i e l l a p n e u m o n i a e NIH I I I

500

M i c r o c o c c u s p y o g e n e s v a r . a u r e u s 20 9 P

312

M y c o b a c t e r Turn p h l e i

125

Mycobacterium smegmatis

125

Mycobacterium t u b e r c u l o s i s

D.T.

31

Mycobacterium t u b e r c u l o s i s

H37

31

Mycobacterium t u b e r c u l o s i s

H37Rv

31

P r o t e u s s p . OXK 11+11+
Ps e udomona s a e r u g i n o s a R-P-CC

500
312

Salmonella typhimurium 9

500

S a r c i n i a l u t e a PCI 1001
Shigella paradysenteriae

300 9

1-56
312

S t r e p t o c o c c u s h e m o l y t i c u s 286

39

Xanthomonas p h a s e o l i

78

5i+
Since

the

a l k a l o i d was n o t

preparations
wa s

less

the

than

serves for

on e o f

inhibitory action

that

of

of

purposes,

impurities

provided

Gottshall

Michigan Department

capsine

cent

out

sterile

sterile

of

1,

0.1

2,

ml.

cavity

weight

of p r o p y l e n e

base

of

0 .9

propylene
per

cent

injected

either

for

100 mg.

glycol

and

of

strain,

mice were

the

slowly
solution.
into

the

in amounts

injected with

glycol.

i t wa s n e c e s s a r y

mg./kg.

Y.

solano-

sodium c h l o r i d e

route

only 50-100 mg./kg.

so 1 a n o c a p s i n e wa s t o l e r a t e d ,

The

and

s u b c u t a n e o u s l y and

The c o n t r o l

intraperitoneal

succumbed.

b y D r . R.

so 1a n o c a p s i n e

o f w h i t e mi c e' , W e b s t e r

1+, a n d 6 mg.

By t h e

a possible

th e D i v i s i o n of L a b o r a t o r i e s ,

w a s made b y d i s s o l v i n g

The c o m p o u n d s w e r e
peritoneal

table

on So 1 a n o c ap s i ne

s o l u t i o n of b o th

in 2 m l . of

adding 8 m l . of

at

the

of Health'*.

hydrochloride

c o mp o u n d

that

d e t e r m i n a t i o n s were d i r e c t e d

and c a r r i e d

A 1 per

However,

i s g i v e n due c o n s i d e r a t i o n .

L|_. T o x i c i t y S t u d i e s
toxicity

a g a i n s t M. t u b e r c u l o s i s

other preparations.

comparative

enhancing e f f e c t

The

the most h i g h l y p u r i f i e d

lethal

to

inject

dose

whereas with the

200 m g . / k g .

by s u b c u t a n e o u s

so 1 a n o c a p s i n e

or

of body

before

the

free
animal

i n j e c t i o n wa s 300

so 1 a n o c a p s i n e

hydrochlo-

r ide .

“ The a u t h o r w o u l d l i k e t o t h a n k M r s .
conducting the t o x i c i t y determ inations.

S h irle y Geis for

65
After

injection

of

as

little

t h e mo u s e b e c a m e v e r y n e r v o u s .
lation

short

strating
the

posterior

limbs were

m i n u t e s t h e mo u s e
shallow.
three

Th e

days

tered.

collapse

sta rte d with

almost

the

so 1 a n o c ap s i n e

immobile.

if

animal

after

animal

After

inocu­
d e mo n ­

15 m i n u t e s

Following

30-60

b r e a t h i n g wa s r a p i d

u s u a l l y remained c o ll a p s e d

and t h e n e x p i r e d ,

Otherwise,

the

between s e i z u r e s .

usually collapsed;

animal

of

From 9 - 1 5 m i n u t e s

jerky convulsions

complete

a s 0 . 2 5 nig.

lethal

and

f r o m one t o

dose had b e e n a d m i n i s ­

recovered

a nd a p p e a r e d n o r m a l

a nd a c t i v e .
I n ma n y c a s e s
into

t h e mice

a c r u s t y eczema over

to the back.

developed

the

A scabby sore

entire

sores,

which o f t e n

body or e l s e

u s u a lly appeared at

it

the

turned

localized
site

of

inject ion.
5. A t t e m p t s
Ail

alkaloids

to

isolated

been g l y c o s i d e s w ith
were

to

attach

among s i m i l a r

the

compounds
that

alkaloid

Two p l a u s i b l e

the

an A l k a l o i d a l

so f a r

reasons

from the

e x c e p t i o n of

any s i g n i f i c a n c e

one woul d c o n c l u d e
and t h a t

Isolate

to

isolated

the

Glycoside
genus Solanum have

so 1a n o c a p s i n e .
genetic

in the p l a n t

for

an a l k a l o i d

finding
Either

a very active

catalyses

of the

c o mp oun d a t

vesting

the

plant

or

the

species,

so 1a n o c a p s i n e p r o b a b l y wa s an a r t i f a c t

was p r e s e n t

hydrolysis

relationships

from c l o s e l y r e l a t e d

c o s i d e may b e p r o p o s e d :
the

I f one

glycoside

is

as a g l y c o s i d e .
and n o t

a gly­

glycosidase

the

time

of h a r ­

so l a b i l e

that

the

56
we a k a c i d

used

in the

is o la tio n procedure w ill

hydrolyze

the

compound.
a.. P o s s i b i l i t y
tions

supported

of

the

a g l y c o s i d e . - The f o l l o w i n g

th eo ry of

glycoside

in th e p l a n t :

difficult

to p u rify

s t a n t s were
Different
weak,
for

identical

batches

furfural

tive

leaf

cases

the

after

crude

test

ring

at

Paper

root

gave upon p a p e r
aniline

a blank

root

alkaloid

Molisch t e s t .

5,

20,

same

the

a reddish

of a ra b in o s e .
assumption th at

the

as a g l y c o s i d e ,

h y d r o l y s i s wa s a t t e m p t e d .
A solution

0.1 M s u l f u r i c
15,

gave,

1+) The

hydrogen p h t h a l a t e ,

c h r o m a t o g r a p h y . - Un d e r

10,

Inci­

c h r o m a t o g r a p h y and d e t e c t i o n

so 1 a n o c a p s i n e wa s p a r t l y p r e s e n t

i n 0 . 5 ml . o f

test.

3) The m o t h e r

comparable with the

and e n z y m a t i c

given a p o s i ­

w a t e r wa s f o u n d t o

interface.

of the

a positive

Acid h y d r o l y s i s :

0,

distilled

the

B a t c h e s of

a negative Molisch t e s t .

wa s c o m p a r e d w i t h

te s t with

a

characteristic

substances.

gave

2)

u s u a l l y gave

i n one c a s e h a v e

so 1a n o c a p s i n e

spots with

both acid

After

only

con­

so 1 a n o c a p s i n e .

(l+3a), a t e s t

s po t w i t h an R ^ - v a l u e
b.

found f o r

test

concentration,

sugar

the physical

Molisch

purple

liquor

that

so 1 a n o c a p s i n e

from r e c r y s t a l l i z a t i o n

mother
of

those

as a

a l k a l o i d wa s v e r y

root

the b la n k

show a f a i n t
liquor

with

a n d f u r f u r a l - y i e 1d i n g

Pure

dentally,

isolated

such an e x t e n t

so 1 a n o c a p s i n e

test.

In a l l

1) The

occurring

of crude

but p o s itiv e

crude

to

so 1 a n o c a p s i n e

observa­

1+0,

o f i| . 8

mg.

of crude

a c i d wa s r e f l u x e d
and

130 m i n u t e s

alkaloid

on s t e a m b a t h .

5-pl • a liq u o ts

57

were

removed w i t h

Wh a t ma n n o .

a micropipet

1 filter

paper.

and d e p o s i t e d

on a s h e e t

The c h r o m a t o g r a m wa s d e v e l o p e d

with

1- b u t a n o 1- e t h a n o l - w a t e r ( 8 0 : 2 0 : 2 0 ) and a s i n g l e

spot

of Rj,-value

phthalate
minutes

0 . 2 5 - 0 . 2 6 wa s d e t e c t e d w i t h

reagent

gave

the

In a n o t h e r
dissolved

(23).

and

Enzymatic

hydrolysis:

crystalline

the

on p a p e r

hydrolysis

five

alone

but

a nd

e x p e r i m e n t s were

a nd Co mp a n y T a k a - D i a s t a s e

in

c h r o m a t o g r a p h y on p a p e r
Neither

for

d i d m e t h a n o l y s i s of

Table XII
in the

leaf

so 1a n o c a p s i n e , a nd

were e a c h c h r o m a t o g r a p h e d .

of e th y l

(1+9), w a s u s e d .

pounds developed

for

test

any such r e s u l t .

so 1a n o c a p s i n e

(85).

1 0 0 ° C.

Other

acetate-glacial

All

15 p e r

cent

of

records

As

acetic

sol­

acid-

a 85 p e r

four m a t e r i a l s produced

same R „ - v a l u e wh e n d e t e c t e d w i t h
f

reagent

at

130

a l k a l o i d wa s

in a small

arabinose.

w a t e r ( 3 : 1 : 3 ) t o w h i c h was a dded
ethanol

crude

acid

for

A g a i n a s u g a r wa s d e t e c t e d

so 1a n o c a p s i n e , c r u d e

upper phase

a n i l i ne h y d r o g e n

a pentose.

with

any s u c c e s s .

t h e c o mp o u n d p r o d u c e
Crude r o o t

Davis

solutions;

s u g a r wa s w i t h o u t

the

reddish

hydrolyzed

for

spotted

pentoses.

identifiable

buffered

the

of

and h e a t e d

A l i q u o t s were

conducted w ith Parke,

vent,

expected

10 mg.

sealed

with d iffe re n t

i t was most

neutral

aliquot

in 1 m l . of N h y d r o c h l o r i c

in an ov e n .

together

spot

experiment

t ube wh i c h t h e n was
hours

Only the

reddish

of

cent

s p o t s of

a modified Dragendorff

the R e v a l u e s

above m e nt i on ed

for

and o t h e r

the

c om­

solvent o.

58
TABLE X I I
R f -VALUES FOR SOLANOCAPS INE, CRUDE LEAF
ALKALOID, AND CRUDE ROOT ALKALOID
C r y s t a l 1 ine
So 1a n o ­
cap s ine

So 1v e n t
1- b u t a n o 1- e t h a n o 1 - w a t e r
(8 0 : 2 0 : 2 0 )
1)

no

Amorphous
Crude Leaf
A l k a l o id

spot

Amorphous
Crude Root
A l k a l o id
no s p o t

0.25

Up p e r p h a s e o f e t h y l
acetate-acetic acidwater ( 3 : 1 : 3 ) to which
is added
of 85%
ethanol
2)

0.57

0 .5 6

0.57

1- b u t a n o 1- e t h a n o 1 p h o s p h a te b u f f e r (pH8.2)
(8 0 : 2 0 : 2 0 )
2)

0. 71;

0.75

0 .7 k

E t h a n o l - N HC1 - w a t e r
( 5 0 : 2 :l|-8 )
2)

0.73

0. 71;

0.73

1) S p o t s d e v e l o p e d w i t h
2)

hydrogen p h th a la te

reagent.

Spots developed with modified Dragendorff reagent.
These

S.

aniline

findings

indicated

p s e u d o c a p s i c u m was t h e

a glycoside.
existence
It

of

so 1 a n o c a p s i n e
stated

so 1 a n i d i n e w e r e p r e s e n t
alcoholic

extracts

solanine,

solanidine,

side by side
alkaloids.

free

Nevertheless,

had b een

in t h e
The

that

alkaloid

alkamine

the

results

in the p l a n t

in t he
i n S.

from the

the

isolated

so 1 a n o c a p s i n e

that

leaves

solanine

solvent

a nd

Therefore,

and s a m p l e s o f

and t o m a t i n e wer e p a p e r

aforementioned

the

as a g l y c o s i d e .

p s e u d o c a p s i cum ( 6 6 ) .
and

a nd n o t

did not d i s c l o s e

literature

roots

from

for

chromatographed
developing

a l k a l o i d s were d e t e c t e d w i t h t h e m o d i f i e d

59

Dragendorff

reagent.

gave o n l y one

spot w ith

and r e - r u n

these

the

front,

liquid

0.08

and 0 . 0 7 ,

each

while

spot

of 0 .5 7 .

as

extracts

Then,

whe n e l u t e d

Solanidine

followed

a nd t o m a t i n e h a d R ^ - v a l u e s o f

Thus

i t wa s f o u n d

given by the

in R ^ - v a l u e

ethanolic

an Rf o f 0 . 2 2 .

solanine

r e s p e c t i v e 1y .

identified

and l e a f

s p o t s had a R

and o n l y a l k a l o i d
wa s

The r o o t

that

root

a nd

that

leaf

t h e one

extracts

given by a pure

sampl e o f

so 1 a n o c ap s i n e .
c.

Isolation

possibility
glycoside

of

a t t e m p t s . - As m e n t i o n e d p r e v i o u s l y ,

a glycosidase

could

not

be e x c l u d e d .

s u c h a n e n z y me w a s a t t e m p t e d
vesting

the p l a n t s .

s o 1a n o c a p s i n e
ether
sides,

is

ethyl

a poor

2)

the

the

day of

1)

stored

dehydrated

this

class

of

har­

the basic

but

since

f o r mo r e h y d r o p h i l i c

time

i n a c t i v a t i o n were

in d r y

glyco­

o f c o mp o u n d s

ice

utilized:

in a b s o l u t e

a nd s t o r a g e

1)

ethanol

and

in deep f r e e z e

i s o l a t i o n wh e n t h e y w e r e b o i l e d w i t h w a t e r .
and t o p s

a nd e a c h p a r t

until

after

Isolating

as a s o lv e n t,

harvested plants

The r o o t s

separated

inactivation

as p o s s i b l e

isolating

of a

( 8 l|_).

immediate f r e e z i n g

until

ether

for

Two m e t h o d s o f e n z y me
Submersion of

Therefore,

as f a s t

solvent

a s t a n d a r d method

was e m p l o y e d

the h y d ro ly sis

The m e t h o d u s e d f o r

utilized

probably

catalyzing

the

for

of

15 f r e s h p l a n t s w e r e

covered with

isolation.

t o p s wa s m a c e r a t e d

absolute

A total

of

ethanol
1LpOO g.

a nd
of

in a War i n g b l e n d e r w i t h

immediately

60

Ilf 1 • o f 2 p e r

cent

acetic

hours with occasional
by f i l t r a t i o n

and

acid

stirring.

the p l a n t

w i t h 5 .J- o f 2 p e r

cent

to

about

ing o v e r n i g h t
collected

acetic

contained

100-ml.

g.

but

from the mother

liquor.

move d a n d d r i e d

at

at

of

the

to

After stand­

each time f o r

the
which

one

green c olored methanol

a c o mp oun d c r y s t a l l i z e d

give

and

wa s r e f l u x e d f i v e

adsorbed r a p i d l y green

f r o m 60 p e r

alkaline

and t h e p r e c i p i t a t e

salts,

The c r y s t a l l i n e

l±5°C.

and f i l t e r e d .

room t e m p e r a t u r e

of methanol,

evaporator

clusters,

recrystallized

2 l| h o u r s

a precipitate.

inorganic

portions

in a f l a s h

needle

sim ilarly re-extracted

for

drying

1+8

The d a r k b r o w n p r e c i p i t a t e ,

During c o n c e n tr a ti o n

extracts

stand for

w e r e made d i s t i n c t l y

After

amounts of

to

s u p e r n a t a n t wa s r e m o v e d

s u p e r n a t a n t wa s d e c a n t e d

weighed 0 .7 6

large

times with

fine

the

acid

to flo c c u la te

on a f i l t e r .

precipitate

hour.

50°C.

The

p u l p was

The c o m b i n e d f i l t r a t e s
heated

and a l l o w e d

20 mg.

impurities

precipitate
of p ro d u ct.

cent methanol

in

wa s r e ­
I t wa s

w i t h n o t much s u c c e s s

in d e c o l o r i z i n g .
The r o o t s
described
yield

of

for

the

isolated

2)

For

harvested,
freeze.
placed

(350g.)

were

tops.

the

in b o i l i n g w a t e r
continued

in t h e

of

in d r y

glycoside
for

same way a s j u s t
agent

and

see T a b l e X I I I .

second p a r t

time of

in t he

amounts of e x t r a c t i o n

immediately placed

At

procedure

For

c o mp o u n d
the

treated

the

experiment

13 p l a n t s w e r e

ice and p r e s e r v e d

extraction

10 m i n u t e s

t h e p l a n t s were

and t h e n

same wa y a s a b o v e .

in deep

isolation

61
Th e v a r i o u s
summarized

data

concerned with

these

experiments

are

in T ab le X I I I .
TABLE X I I I

DATA CONCERNING ATTEMPTED ISOLATION OF ALKALOID GLYCOSIDE
Yield
2 n d T ime
1 s t T ime
of
Crude
We i g h t Mace r a t e d Ma c e r a t e d
Stored
( Dehyw i t h 5% " g l y c o w i t h S%
d r a t e d ) A c e t . A c id A c e t . A c id
s i de"
in

Part
Number
of
of
P l a n t s P 1a n t s

s t er ns
and
6 1 . abs
1e ave s
ethano1
15
r oot s
13

3 1 . ab s
ethano1
deep
freeze

who 1 e
p 1a n t s

Th e t h r e e
of

175-195°C.

which

gave

5.0

1.

0.76

g.

2 0 mg

1.

1 .0

1.

1 .28 g .

61 mg

lip. 0 1 .

5.0

1 . 1 2 . 5 g-

11; . 0 1 .

350 g

3.5

2200 g

compounds

solvents mentioned
all

114-00 g .

isolated

suggested

all

melted

impurities.

as

128 mg

in the board range

Chromatography

i n T a b l e XIV s ho we d t h a t

same a l k a l o i d R ^ . - v a l u e

Yield
of
Cr ude
"glycos i de'

the

three

so 1 a n o c ap s i n e .

If

in t he

c o mp o u n d s
the

com­

pounds were c h r o m a t o g r a p h e d

in I - b u t a n o 1- e t h a n o l - w a t e r (8 0 : 2 0 : 2 0 )

and t h e

aniline

spots detected with

c o mp o u n d

isolated

from the p l a n t

from the f r o z e n p l a n t s
The r e s u l t
possible
by t h e
sides

to

of

isolate

conventional
it

s h o we d

these

tops

spots

ex pe r iments

an a l k a l o i d

and t h e

strongly that

c o mp o u n d

only the
isolated

of D - g l u c o s e .
indicate

glycoside

method used f o r

suggests very

hydrogen p h t h a l a t e ,

that

f r o m S.

isolating

it

is not

pseudocapsicum

glycosides.

Be­

so 1 a n o c a p s i n e p r o b a b l y d o e s

62

not e x i s t

in th e p l a n t

loid,

since

yield

a glycoside.

as

inactivation

a glycoside,
of

a possible

but

as a genuine

glycosidase

alka­

did not

TABLE XIV
R f -VALUES OF COMPOUNDS ISOLATED BY A STANDARD
PROCEDURE FOR ALKALOIDAL GLYCOSIDES

So 1v e n t

Solanocap s ine

1- b u t a n o l - e t h a n o l w a t er (8 0 : 2 0 : 2 0 )

1)

R^ - V a l u e of ;
Compo und Compound Compound
Isolated
Isolated
Isolated
f r om
f r om
f r om
Top s
Who 1 e
Roots
P 1a n t

no s p o t

0 .2 0

no s p o t

0.19

Up p e r p h a s e o f e t h y l
acetate-acetic acidwater ( 3 : 1 :3 ) to which
is added
of 85%
ethanol
2)

0.57

0.57

0.5 6

0.57

1- b u t a n o 1- e t h a n o 1 p h o s p h a t e b u f f e r (pH
8 . 2 ) (8 0 : 2 0 : 2 0 )
2)

0.71+

0.71+

0.75

0.74

Ethanol-N HCl-water
( 50 : 2 : 1+8 )
2)

0.73

0.71+

0 .71+

0.73

an i 1 ine h y d r o g e n p h t h a 1a t e r e a g e n t .
2) S p o t s d e v e l o p e d w i t h m o d i f i e d D r a g e n d o r f f r e a g e n t .
1) S p o t s d e v e l o p e d w i t h

6 . P r i n c i p 1 e w i t h A n t i b a c t e r i a l A c t iv i t y
A g a i n s t D. p n e u m o n i a e

In s e v e r a l
that

the

instances

antibacterial

during

activity

this

i t wa s o b s e r v e d

a g a i n s t D. p n e u m o n i a e

t o be a s s o c i a t e d w i t h

the

colored

were f o u n d

the

fractions

i n some o f

study

impurities.

s e e me d

Su c h e x a m p l e s

o f an e t h a n o l i c

extract

63

from r o o t s

c h r o m a t o g r a p h e d on a l u m i n a

the p u r i f i c a t i o n
so 1a n o c a p s i n e
of c o lo r e d

of

crude

fraction

Twenty grams o f
of 80 p e r

obtained e x tra c t
a c i d wa s a d d e d ,
ml . p o r t i o n s

cent

ethanol

ether.

(solution

II).

ether

hours with

extractor.

ml.,

10 m l .

The

of a c e t i c

twice with 200fractions

evaporator

to

give

a

of a b s o l u t e

ethanol

t wo p r e p a r a t i o n s w e r e

then te s te d

for

ethanol

results

that

other

extraction

principle

a f r a c t i o n wa s o b t a i n ­

a g a i n s t D. p n e u m o n i a s .

principle

of

i n T a b l e XV.

antibacterial

since

activity

by an a t t e m p t

(solution

shown

another

is p r e s e n t ,

antibacterial

T h i s wa s t e s t e d

four

in ^0 m l .

s h o we d a c t i v i t y

the

to

phase.

T h i s r e s i d u e was p a r t l y d i s ­

cent

a c t i v i t y with

that

for

to prevent

and t h e

solanocapsine

high

the ether

The c o m b i n e d e t h e r

was d i s s o l v e d

The

by e t h e r

I),

T a b l e XV i n d i c a t e s

ed w h i c h o n l y

into

in a S o x h l e t

of m a t e r i a l .

residue

a direct

i t wa s f r e e d

e x t r a c t made a c i d i c

in a f l a s h

i n 50 itU . o f 8 0 p e r

suggested

as

s o l u t i o n wa s e x t r a c t e d

to dryness

remaining

fairly

root

wa s c o n c e n t r a t e d

o f 2 1 0 mg.

besides

in

a c t i v i t y of

this principle

r o o t s were r e f l u x e d

of e t h y l

antibacterial

isolate

going over

and t h e

were e v a p o r a t e d

solved

to

of an e t h a n o l i c

alkaloid

yield

the

Also

impurities.

extraction

25*0 m l .

solanocapsine,

1|1 ) .

a g a i n s t D, p n e u m o n i a e d e c r e a s e d

An a t t e m p t w a s made

the

(page

to
the

of
wa s

solutions
in f a c t

isolate

I a nd

The
II

organophi1i c .

this principle

r o o t powder.

by

61+

TABLE XV
ANTIBACTERIAL ACTI VI TY OF FRACT IONS DURING ISOLATION
OF PRI NCI PLE ACTIVE AGAINST D. PNEUMONIAE
^ / m l . I nh ib i t s Gr owt h o f :
D. p n e u m o n i a e
M. t u b e r c u l o s i s

- Fraction

ethanolic

root

extract

720

255

so 1u t i o n

I

11

i nac t ive

sol ut ion

11

22

i n a c t ive

19

8

water phase
(m o t h e r 1 i q u o r )

Four hundred
1 . 6 1.

of ethyl

( 61|) f o r

grams o f

ether.

continuous

l i q u i d was u s e d .
outlet

tube

An a p p a r a t u s

extraction

A plug

of the

d r y r o o t p o w d e r wa s e x t r a c t e d w i t h

of

as d e s c r i b e d by Morton

of a l i q u i d w i t h

g l a s s wo o l wa s

column and

a heavier

inserted

i t wa s t h e n f i l l e d

into

the

w ith the

r o o t p o w d e r , w h i c h c o u l d be c o n t i n u o u s l y e x t r a c t e d w i t h e t h e r .
By t h i s

p r o c e s s 550 ml.

wa s o b t a i n e d w h i c h ,
did not
If
anol,

of

wh e n t e s t e d

for

the

p o w d e r w a s r e - e x t r a c t e d w i t h 80 p e r

a s o l u t i o n was o b t a i n e d ,
a nd M.

corresponded very c lo s e ly
countered

during

Fr o m t h i s

isolation

experiment

can e x t r a c t

the

a g a i n s t D_. p n e umon i a e .

w h i c h had h i g h

tuberculosis.
to
of
it

the

These

antibacterial

solanocapsine
c a n be

principle

solution

activity,

inhibiting

root

effect

antibacterial

show a n y

b o t h D. p n e u m o n i a e

ether

a strongly yellow colored

active

activity
activity

eth­

against
xe s u l t s

activities

from the

seen t h a t

cent

en­

roots.

although ethyl

a g a i n s t D.

p n e umon i_ae

65

fr om an e t h a n o l i c
plant m aterial

extract,

directly.

has not been f u r t h e r

it
The

c a n n o t be e x t r a c t e d

from the

significance

observation

examined.

7. A n t i t u b e r c u l a r A c t i v i t y
Since

so 1a n o c a p s i n e

wa s d e c i d e d
plants

test

belonging

a.
lated

to

t um M i l l .

certain

leaves

fungi.

of

is

the

ant itu b e rc u la r

their

and f o u n d

Solanine

(57)

and

found

with

its

is e s p e c i a l l y high

have been a l l o w e d

to

f r o m So 1a n a c e a e

are found

e r a t e d w i t h 2 1.
and a l l o w e d

to

centrifuged

in an

method f o r

Green p o ta to

stand for

is

The h i g h e s t

sprouts

the p o t a t o

i s o l a t i o n method f o r

of 2 per

solanidine

(52) .

tubers

in d a y l i g h t .

conventional
(93).

M.

i s an a l k a l o i d

in p o t a t o
if

l ow

against

against

S o l anurn t u b e r o sum L.

The c o n c e n t r a t i o n

the

iso­

inactive.

aglycone

solanine

of

activity

i t wa s t e s t e d

s o l a n id i n e . - Solanine

sprout

glycoside

to have on l y v e r y

but higher

c o n e e n t r a t I ons of

- The

it

t o be c o m p l e t e l y

in the p o t a t o p l a n t ,

ification

it

g l y c o s i d e s of o t h e r

an a l k a l o i d a l

experiment

together

Isolation.

activity

tomato p l a n t Lycoper s icon e s c u l e n -

most b a c t e r i a ,
In t h i s

gly co s id e which
present

or

Solanum A l k a l o i d s

to S o la n a c e a e .

against

tuberculosis
b.

alkaloids

Ma a n d F o n t a i n e

activity

of Other

exhibited

T o ma t i n e . - T o m a t i n e

from the

of t h i s

cent

I4.8 h o u r s .

International

alkaloid

sprouts

acetic

acid

The

s o l a n i n e wa s a mod ­
glycosides

( 9 7 0 g. ) wa s m a c ­
in a Waring b l e n d e r

s u s p e n s i o n wa s t h e n

centrifuge

(size

2)

and t h e

66
s u p e r n a t a n t was f i l t e r e d
cel.

w ith the

The r e m a i n i n g p l a n t

with

750 m l .

of 2 per

aid

of a l i t t l e

r e s i d u e was r e - e x t r a c t e d ,

cent

acetic

acid.

After

s u s p e n s i o n was c e n t r i f u g e d

a nd f i l t e r e d

filtrate

the

was combined w i t h

Concentrated
br own f i l t r a t e s
the

turbidity

solution

of

on a f i l t e r ,
droxide,

1 .)

the

until

After

a short

2l_i_ h o u r s

and d r i e d

inorganic

time

the

of 5 per

The y i e l d

a nd

By h e a t i n g

the

alkaloid

wa s c o l l e c t e d

cent

ammoni um h y ­
at

of crude

65°C.

i n an

solanine

and

The p r o d u c t wa s g r e a t l y c o n t a m i n a t e d

salts.

P u r i f i c a t i o n . - The
idine

is based

ethyl

ether,

ble.

Therefore,

upon t h e

whereas
the

d i n e wa s e x t r a c t e d

separation
fact

that

the basic

solanine

solanine

aglycone

3 . 6 g. m i x t u r e

three

of

of

After

evaporation

of crude,

s l i g h t l y brownish colored
from a c e to n e

(Lit.

217°C.

[93]).

insoluble

solanine

is

to dryness

the

over

solu­

of e t h y l

anhydrous

sodium

t h e y i e l d wa s 30 mg.

solanidine.

c o mp o u n d m e l t e d a t

No f u r t h e r

in

a nd s o l a n i ­

times with 200- m l . p o r t io n s

sulfate.

tallization

is

from s o l a n ­

solanidine

e t h e r w h i c h we r e c o m b i n e d and d e h y d r a t e d

value

1 ight

remained

crude

over phosphorus pentoxide

s o l a n i d i n e was 3 . 6 g.

the

and t h e

to the

a mmo n i a o d o r

the p r e c i p i t a t e

wa s h e d w i t h 50 ml .

Abderhalden drying p i s t o l .

with

the

time

filtrate.

s o l u t i o n wa s p e r m a n e n t .

to 50- 70°C. for

flocculated.

first

as b e f o r e ,

super-

this

21; h o u r s

ammoni um h y d r o x i d e wa s a d d e d

(3.5

Hyflo

Upon r e c r y s ­
2 0 i ; - 2 0 7 C.

a t t e m p t wa s ma de t o

purify

solanidine.

activity

The c o mp o u n d w a s t e s t e d

and f o u n d

t o be

The r e m a i n i n g
five

times,

cent ethanol.
for

The

for

activity
to

c o n c e n t r a t ion the

solanine

purification

the

w i t h 250 ml.

residue

(1.3

in a f l a s h

c ry s ta l1ized.

g.)

o f 80 p e r
wa s t e s t e d

value

cent

ethanol

285-287°C.

for

During

The y i e l d

the

[ 9 3 ] ).

s o l a n i n e wa s t e s t e d

fractions

evaporator.

of m e l t i n g p o i n t 255~260°C.

f ro m 80 p e r

(Lit.

s o l a n i n e wa s r e f l u x e d

and t h e combi ned e t h a n o l

250 ml .

s o l a n i n e w a s 2 8 0 mg.

at 278-280°C.

crude

o ne h o u r ,

were c o n c e n t r a t e d

crystallization

of

undissolved

antitubercular

antitubercular

inactive.

residue

each time

for

of crude

After

re­

c ompoun d m e l t e d
Without

further

antitubercular

ac­

tivity.
The r e s u l t s
activity
(25

^ / ml . ) .

presence
not

against

were
D.

s how a n y

Crude

This

confirmation
therefore

The

s h o we d we a k a n t i b a c t e r i a l
a n d M. t u b e r c uu ll oo ss ii s

(125

s h o we d no a n t i b a c t e r i a l
insoluble

activity

residue obtained

in the

above

alkaloid

has been recorded

information
since

p r e p a r a t i o n f r o m S.

the

a possibility

is

in e a r l y

carolinense. -

as c o n t a i n i n g

literature

solanine

and r e q u i r e s

i d e n t i f i c a t i o n wa s s u p e r f i c i a l
of

other

alkaloidal

a nd

g l y c o s i d e s can

n o t be e x c l u d e d .
A 1000 g.
and

did

inhibitoryeffect.

Sol anurn c a r o l i n e n s e
(90).

solanine

p n e u mo n i a e

Solanine

of b l o o d .

c.

that

sample o f d r i e d

s t e ms was m a c e r a t e d f o r

a n d g r o u n d ( 2 0 me s h )

i;8 h o u r s

in a l a r g e

leaves

glass

68

cylindrical
acetic
the

jar

acid.

filtrate

a nd t h e

with

The p l a n t

alkaloid.

a short

After

a filter,

the
time

2 l| h o u r s

at

a l k a l o i d were

base.

tested

at

50-70°C.

tuberculosis

no f u r t h e r

The r e s u l t s
mentioned

attempt
obtained
are

to give

a n t i t u b e r c u 1a r

32 mg.

liquor

from the

crude
on

11 g.

the crude
The m o t h e r

inhibited
the

growth
crude

In v i e w o f t h i s

a l k a l o i d wa s ma d e .

testing

summarized

of

and

activity.

inactive.

the

ammoni um h y d r o x ­

1 :5 ? whereas

to p u r i f y the

the

a yield

of dry m atter
of

Heating

wa s c o l l e c t e d

cent

the mother

in a d i l u t i o n

alkaloids

glacial

odor p e r s i s t e d

flocculated

of 5 per

a l k a l o i d p r o d u c t wa s c o m p l e t e l y
result

the

the p r e c i p i t a t e

Both

for

liquor which c o n ta in e d
o f M.

until

room t e m p e r a t u r e

alkaloid

of

s o l u t i o n wa s p e r m a n e n t .

washed w i t h 200 ml.

i de , a n d d r i e d
of crude

of

and 600 ml.

r e s i d u e was removed by f i l t r a t i o n ,

w a s made a m m o n i a c a l

turbidity

solution for

30 1 . o f w a t e r

of

the

above

i n T a b l e XVI .

TABLE XVI
ANTIBACTERIAL ACTI VI TY OF D IFFERENT SOLANUM ALKALOIDS
X?ml . I n h i b i t s t h e G r o w t h o f :
D. p n e umon i ae
M. t u b e r c u l o s i s
On B l o o d
Agar
On A g a r

Alkaloid

Tom at in e
So 1 a n i n e

i n a c t i ve

25

i n a c t ive

i n a c t i ve

i n a c t i ve

i n a c t i ve

i n a c t ive

i n a c t i ve

i n a c t i ve

125

Sol an id i ne
Alkaloid p reparation
S . carolinense

i n a c t i ve

i n a c t ive

from

69

CHEMICAL STUDIES ON SOLANOCAPSINE
8 .

Physical

n . Sol a noca p s ine . - F i v e
sine

dihydrochloride

(1:1).

By a d d i t i o n

precipitated.

The y i e l d

recrystallization

lized

as p l a t e l e t s

(Lit.

value

pressure

over

of crude

solanocap­

o f e t h a n o 1- w a t e r
solanocapsine

s o l a n o c a p s i n e wa s

f r o m e t h a n o 1- w a t e r

(1:1)

370 mg.

it

crystal­

which melted w ith decomposition at 2l5-2 l6 °C .

2l6-217°C.

analysis

i n 10 m l .

6 N ammoni um h y d r o x i d e

After

For

h u n d r e d m i 11 i g r arris o f

was d i s s o l v e d
of

Constants

the

[78],

222°C.

[ 6 ]).

c o mp o u n d wa s d r i e d l_j_8 h o u r s

phosphorus pentoxide

and p a r a f f i n

at

at

17 mm.

r oom

temper a t u r e .
An a l . C a l c ' d

for

C ^ y H ^ N ^ O ^ *HpO:

N,

6 . 2 I4. %

Found:

N,

6.20,

D e te rm in a tio n of water
dried

crystallization:

in an A b d e r h a l d e n d r y i n g p i s t o l

pressure

over phosphorus p e n to x id e .

r e f l ux ing 1 - b u t a n o l
mg.)

lost

cent

(calculated

posure

2 5 . 6 mg.

to

air

the

(b.p.

for

72 h o u r s

which c o rre sp o n d s

substance

at

to

its

( 6 I4.2 . 3

3.98 per

cent) .

regained

2 mm.

wa s h e a t e d by

The c o mp o u n d

1 m o l e o f w a t e r I4..OI p e r

anhydrous

The c o mp o u n d wa s

The p i s t o l

1I7-H8°C.).

by d r y i n g ,

for

6.29 % (Kjeldahl)

On e x ­

original

we i g h t .
Optical
methanol

rotation:

A solution

s h o we d a r o t a t i o n

Calculated

specific

o f 0 . 2 0 6 mg.

of + 2.061

rotation

i n 5 ml ■

i n a 20 cm.

[ct ] 2^ = + 2 5 . 0 1 °.

tube.

70
R ast's
19k., 2 mg.
ponding

molecular weight

camphor

to

depressed

7.6°C.,

Neutralization
dissolved

ing t o

ethanol

12.05 ml.

a molecular

to

5.71+- m l .

( 8 . 6 mg.)

a molecular

oxidation

hydroxide

(oxidation

Calculated

for

cribed

groups

used

after

attached
oxidation

recovered

and

t i m e wa s
3 methyl

to carbon:
a nd t i t r a t i o n

1+.80 m l .

( 7 . 1 mg. )

0 . 0 1 3 N sodium

six hours).
10.05 %

croups:

oxidation following
the hydroxyl

identified

correspond­

Solanocapsine

and d i s t i l l a t i o n

oxidize

( 6 3 . 5 mg.)

0.0102 N h ydrochloric

10.28,

did not

o f 1+81 .

o f 1+8 2 .

Found:
Oppenauer

the m e l t i n g

weight

Solanocapsine

0.0 103 N sodium h y d r o x i d e .

used a f t e r

depressed

a n d 1+0 m l .

D e t e r m i n a t i o n of methyl
So 1 a n o c a p s i n e

5 . 1 °C . , c o r r e s ­

0 .0 12 0 N sodium h y d ro x i d e

we i g h t

in

S e v e n t e e n and f o u r -

camphor

equivalent:

i n 10 m l .

a c i d consumed

o f 1+56.

i n 1 9 . 3 mg.

corresponding

1 1 . 3 mg.

the m elting point

a molecular weight

tenths milligrams
point

determination:

10.1+2 %

the procedure
group.

later

des­

S o l a n o c a p s i n e wa s

through melting point

and

infrared

spectrum.
One m i l l i g r a m
ultraviolet

part

of

in met han ol
the

s h o we d no a b s o r p t i o n

in the

spectrum.

No u n s a t u r a t i o n wa s d e t e c t e d w i t h p o t a s s i u m p e r m a n g a n a t e
(8 0 ) or bromine

(8 1 ).

Solanocapsine
a nd m i x e d w i t h

(10 mg.)

10 m l .

dissolved

of a 1 per

cent

i n 10 m l .

of ethanol

digitonin

solution

in

Solanocapsine

(Nujol

Mul l )

o

to

jc:
cn

C
0)
>

o
o

o

o

o

CO

u o i s s j uisuB j x T-uao uod

o

OJ

cd

Figure

5.

£/)
c
o
fo

Infrared

Spectrum

of

GO

o
OJ

1|0 ml.

X
o

o

C\J
i—
H

ethanol

and

o
o

o
c

to

cx

dissolved

in

10 ml.

GO

cd

O
in
X
o
o
o

cd
>

u3
CJ
d
o
X-l
+->
cd
x

GO

fcvO
Ci)
X
G3
CD
»H

X

o
a

CO

CXI

mg. of

o
c

(63.5

Oj

solanocapsine

cd

O

N H C i)

0.01020

72

73
90 p e r

cent ethanol

remained c le a r ,

even a f t e r

10 h o u r s o f

stand i n g .
No c o n s u m p t i o n o f p e r i o d i c
10 h o u r s .

a c i d wa s o b s e r v e d ,

The o x i d a t i o n wa s a t t e m p t e d

at

even a f t e r

3 different

hydrogen

ion c o n c e n t r a t i o n s :
pH 2 :
a nd

S o l a n o c a p s i n e (20 mg . )

10 m l . 0 . 0 2 1 6 m o l a r p e r i o d i c
pH

2 ml.

pH 8 :

buffer,

S o l a n o c a p s i n e ( ll|. m g . )
and 7 m l .

Solanocapsine

of c r u d e

dissolved

dissolved

32U°
Before

17 mm.

i n 10

ml. e t h a n o l ,

i n 10

0.0216 molar p e r i o d i c

Th e y i e l d

hydrochloride

ml. e t h a n o l ,
acid.

of pure

wa s r e c r y s t a l l i z e d

solanocapsine

wa s 105 r ag. a nd d i d n o t m e l t b e l o w 3 0 0 ° C .
[ 7 8 ],

ethanol

d i h y d r o c h l o r i d e . - Two h u n d r e d m i l l i g r a m s

solanocapsine

from w a t e r .

ml.

a n d 10 m l . 0 . 0 2 1 6 m o l a r p e r i o d i c a c i d .

pH 9 b u f f e r
b.

i n 10

acid.

S o l a n o c a p s i n e (20 mg.)

pH 8

6 ml.

dissolved

twice

dihydrochloride

(Lit.

value >300°C.

[6]).
analysis

pressure

the

c o mp o u n d wa s d r i e d

over phosphorus pentoxide

for

JL|_8 h o u r s

and p a r a f f i n

at

at

r oom t e m p e r a t u r e .
A n a l .C a l c ' d

for

Found:
D eterm ination of water
c o mp o u n d wa s d r i e d
a t 2mm.

pressure

lost

9.66;

N,

5*37 %

C, 62.27?* H,

9. J+0;

N,

5-59 %

62.17;

crystallization:

A sample of t h e

in an A b d e r h a l d e n d r y i n g p i s t o l

over phosphorous p entoxide.

h e a te d by r e f l u x i n g
(576.8 mg.)

H,

C p y H ^ N ^ O ^ • 2HC1 ' H ^ O : C,

1-butanol

1 9 . if mg.

(b.p.

by d r y i n g ,

for

72 h o u r s

The p i s t o l

117-li8°C.).
corresponding

wa s

The c o mp o u n d
to

3.36 per

7*

in

d
o
c.
o
•’H

CO

a
o

CD
>
(d

o
o

CO

o

vO

o

u o j s s juisire

o

ji

fuao

o

C\J

o
I nf r ar e d

Spectrum

of

Solanocapsine

D I h y d r o c h 1oride

o

Figure

(Nujol

Mul l )

/d-

c e nt ,

(calculated

posure

to

air

for

the

1 mole of w a t e r

anhydrous

3.36 per

substance

cent) .

regained

its

On e x ­

original

we i g h t .
As w i t h
part

of the

solanocapsine

no a b s o r p t i o n

in the

ultraviolet

s p e c t r u m wa s o b s e r v e d .
9. R e a c t i o n w i t h N i t r o u s Acid

a.

N i t r o s o s o l a n o c ap s i n e . - T h i s d e r i v a t i v e wa s p r e p a r e d

following

the

procedure

When a s o l u t i o n

of

a c i d was t r e a t e d
mg.),

110 mg.

with

only a slig h t

of

from a b s o l u t e

ethanol

methanol

Unsaturation
a c e t i c ac i d

double

of

wa s f o r m e d .

it melted

at

(16.9

But a d d i t i o n of

produced

19i j . - 196°C.

s p e c t r u m o f 0 . 2 5 mg.

s h o we d a maxi mum a t

a copious
R e c r y s t a l 1 ized
(Lit.

value

n i t r o s o so 1a n o c a p -

355 mp ( l o g E 2 . 1 | 2 ) .

c o u l d be d e t e c t e d w i t h b r o m i n e

in g l a c i a l

One a n d o n e - h a l f m i l l i g r a m o f t h e

should t h e o r e t i c a l l y

bond).

I t was f o u n d

in 7 ml.

nitrogen,

absorb

that

ami no n i t r o g e n

( 2 1 . 3 mg.)

1 . 3 0 0 mg.

sodium n i t r i t e

(8 1 ) .

Va n S l y k e
c ap s i n e

in 6 N a c e t i c

o f c omp oun d wa s 8 0 mg.

Microhydrogenation:
c o mp o u n d

of

( 6 ).

[78]).

The u l t r a v i o l e t
sine/ml.

solanocapsine

sodium n i t r i t e

The y i e l d

200°C.

a nd F r a e n k e 1- C o n r a t

1 equivalent

precipitate.

[ 6 ],

of

precipitate

a second e q u i v a l e n t

19I+0 C.

of Barger

of

71

75 p i .
pi-

of hydrogen

wa s a b s o r b e d .

determination:
6 N acetic

corresponding

(1

acid

N i t r o s o so 1a n o g ave e v o l u t ion

t o 0 . 9 9 mole of n i t r o g e n .

100

CO

o

o

uoj ssjuisue jx

1 .1 1 3 3

o

J3 j
Infrared

Spectrum

of

Nitrososolonocapsine

CO

8.

in M i c r o n s

o

Fi gur e

Wavel engt h

(Nuj ol

Mull)

76

v0

O

■

"LA

O

•

o

•

o

CO

CVj

•

*

Aoueqaosqv

o

•—<

o

*

230
2^0
270
290
Wa v e l e n g t h
in M i l l i m i c r o n s
Fi gure 9.
U l t r a v i o l e t Spectra

of

330
N i t r o s o s o l a n o c a p s ine

310

and

370

390
ITN i t r o s o d i h y d r o s o i a n o c a p s ine .

3^0

77

78
b.

Digitonide

o f n i t r o s o s o l a n o c a p s i n e . - To a s o l u t i o n

o f 70 m g . n i t r o s o s o l a n o c a p s i n e
20 m l . o f

a 1 per

ethanol.

After

wa s f i l t e r e d ,

digitonin

standing

The d e r i v a t i v e

for

1+8 h o u r s

solution

overnight

washed w i t h w a t e r

digitonide.
a nd d r i e d

cent

i n 20 m l . e t h a n o l

at

the

was mixed

in 80 p e r

cent

copious p r e c i p it a te

and d r i e d

to

g i v e 1+5 mg.

of

was wa s h e d w i t h 5 m l . o f e t h a n o l

17 mm. p r e s s u r e

over phosphorous

pentox i d e .
Anal.

Calc'd

1(£ %

for
Found:

c.

1.90 % (K jeld a h l)

182,

D i h y d r o s o 1a n o c a p s i n e . - A m i x t u r e

capsine

a n d 2 0 0 mg.

acetic

acid

After

of p la tin u m oxide

a n d 5 m l . o f e t h a n o l wa s

of p r e s s u r e

for

12 h o u r s

filtration

by d i s t i l l a t i o n
tor.

N,

of

under

o f 2 0 0 mg.

i n 10 m l .

solano­

of g la c ia l

shaken under

3 atmospheres

i n a n Adams h y d r o g e n a t i o n a p p a r a t u s .
the

catalyst,

the

diminished pressure

The r e s i d u e wa s d i s s o l v e d

s o l v e n t wa s r e mo v e d
in a f l a s h

in e t h a n o l - w a t e r

evapora­

(1:1)

N ammoni um h y d r o x i d e w a s a d d e d f o r

precipitation

yield

I t was r e c r y s t a l l i z e d

of

1 7 0 mg.

ethanol-water
point with

(1:1),

at

in 7 ml.

of

[06 ]

a
from

show a n y d e p r e s s i o n .

s howed a r o t a t i o n

rotation

give

and a m i x e d m e l t i n g

A s o l u t i o n o f 0 . 1 1 2 mg.

ami no n i t r o g e n

( 6 . 7 mg.)

210-212°C.

did not

of methanol

specific

Van S l y k e
c ap s i n e

melted

rotation:

in 5 ml .

Calculated

t h e compound.

solanocapsine

Optical
pound

of

to

a nd 6

of

of

the

1.792

com­
.

= + 1+0°.

determination:

D i h y d r o s o 1a n o -

6 N acetic

g ave e v o l u t ion

acid

79
of 0.791

mg.

of

nitrogen,

corresponding

to

1 . 9 7 mo l e

of n i ­

trogen.
Active

hydrogen determ in atio n :

h y d r o g e n was f o u n d
100°C.

at

Recalculated

s u l t wa s

3 , 614- a c t i v e

room t e m p e r a t u r e

cent

active

a n d O . 8 I4. p e r

t o numbers of a c t i v e

hydrogens

cent
the

at
re­

hydrogens.

T,N i t r o s od i h y d r o s o l a n o c a p s i n e . n - D i h y d r o so 1 a n o c ap 3 i n e

d.
(100 mg.)

was t r e a t e d w i t h n i t r o u s

described

under

acid

n i t r o s o so 1 a n o c a p s i n e

recrystallization
at

O.I 4.2 p e r

from a b s o lu te

in th e

same ma n n e r

and y i e l d e d 8 2 mg.

ethanol

as

After

t h e c o mp oun d m e t l e d

165-168°C.
For

pressure

analysis

the d e riv a tiv e

wa s d r i e d I4.8 h o u r s

over phosphorus pe ntoxide

a nd p a r a f f i n

at

17 mm.

a t r oom t e m ­

perature .
C alcl d for

Anal.

3‘

^2

^ ? 77 - QIp; H,

Found:
Calc’ d for

^ 2 5 ^ 14.0 ^ 6 *

A s o l u t i o n o f 0 . 2 0 mg.
endabsorption

in the

( 79a ) d i d

not

detect

C,

6 8 .67;

H,

9.38 %

^

68.77;

H,

9 . 2 I4. %

compound/ml.

methanol

ultraviolet

Sodium f u s i o n f o l l o w i n g

IO.6I4. %

part

of

the

spectrum.

the microprocedure

any n i t r o g e n

in t h e

theoretically

I t was f o u n d

in g l a c i a l

( 8 1) .

Microhydrogenation:
should

of S c h n e i d e r

c o mp o u n d .

U n s a t u r a t i o n c o u l d be d e t e c t e d w i t h b r o m i n e
ace t ic ac id

s h o we d o n l y

that

One a n d

a b s o r b 93 p i

s e v e n - t e n th s of the
of hydrogen

109 p i w a s a b s o r b e d .

c ompo und

(1 d o u b l e b o n d ) .

100

10.

Infrared

Spectrum

of

CO

CO

o

o

uoj ss

juisub jx

T-tiao J3d

o
OJ
" N i t r o s o d i h y d r o s o l a n o c a p s i n e rT (Nuj ol

o

CD

Fi gur e

M ull).

00

81
A solution
gave w i t h
cent

of

10 mg.

10 m l . o f

ethanol

of

a 1 per

t h e c o mp o u n d
cent

i n 90 p e r

Studies

D i a c e t y l s o l a n o c a p s i n e . - The c o mp o u n d wa s p r e p a r e d

the procedure

lanocapsine
allowed

to

pressure

of Barger

(150 mg.)

a n d F r a e n k e 1- C o n r a t

was d i s s o l v e d

s t a n d w i t h 5 ml.

The y e l l o w

s o l u t i o n was e v a p o r a t e d

a t 90°C.

of

sodium b i c a r b o n a t e

cent

with w ater.

After

e v a p o r a t i o n of
precipitate

the

alumina.

drying

of benzene

Eluate

fractions

composition of the
Fractions

anhydrous
solution

This

after

and

f o r l_p8 h o u r s .

under

diminished

10-ml.

in 2 0 ml

portions

and washed n e u t r a l

sodium s u l f a t e

139 mg.

acetylated

and

of g e l a t i n o u s

a l k a l o i d wa s d i s ­

a n d c h r o m a t o g r a p h e d on 8 . 7

g.

o f 20 m l .

The

were c o l l e c t e d .

ether

( 1 : 9 )*

product.

drying melted

at

of

in T ab le XVII.

1 0 - 1 3 w e r e c o m b i n e d a nd r e p r e c i p i t a t e d

a crystalline

precipiate

over

times with
solution

So­

dissolving

f r a c t i o n s wa s a s f o l l o w s

times from m e t h a n o l - e t h y l
to o b ta in

four

chloroform

was p r o d u c e d .

in 9 ml.

to dryness
after

(6 ).

o f p y r id i n e

anhydride

The b r o w n r e s i d u e ,

was e x t r a c t e d

10 p e r

i n 1|_ m l .

of a c e t i c

of c h l o r o f o r m ,

1 9 6 ° C.

solution

a copious p r e c ip ita te .

a.

solved

of e t h a n o l

digitonin

10. A c e t y l a t i o n

using

i n 10 m l .

three

wa s n o t p o s s i b l e

The a m o r p h o u s ,
198-200°C.

gelatinous

(Lit.

value

193-

[78]).

For

a n a 1y s i s

the

d e r i v a t iv e was d r i e d

over phosp h o ru s p e n to x i d e

and p a r a f f i n

at

at

17 mm. p r e s s u r e

room t e m p e r a t u r e .

82
TABLE XVI I
FRACTIONATION OF DIACETYL SOLANOCAPSINE BY COLUMN
CFfROMATOGRAPHY ON ALUMINA
E 1ua n t

F r a c t ion
1-3

benzene

ll-5

benzene +

Res idue
9 mg.

brown o i l , s o l u b l e
in e t h y l e t h e r

5% c h l o r o f o r m

7 mg.

colorless oil,
s o l u b l e in e t h e r

6-7

b e n z e n e + 10 % c h l o r o f o r m

9 mg .

colorless oil,
s o l u b l e in e t h e r

8-9

benzene + 25% c h i o r o f o r m

5 mg .

colorless oil,
s o l u b l e in e t h e r

10-13

c h l o r o f o r m 4- 1% m e t h a n o l

79 mg.

w h i t e f o a m, i n ­
s o l u b l e in e t h e r

11J.-16

c h l o r o f o r m + 2% m e t h a n o l

6 mg .

Anal . C a l c ' d

for

£ 3 1 ^ 5 0 ^ 2 ^ 11/ ^ 2 ^ *
Found:

D e t e r m in a t io n of
1 .3

equivalents

hydrolysis for

equivalents

greenish o il,
i n s o l u b l e in
e the r

^ > 70.77;

H,9.62 %

C,

H,9.81| %

69.90;

of a c e t y l

groups:

o f 0 , N- a c e t y l ~ ' ~ ) ( 6 . $ mg.

1.9 hour

and d i s t i l l a t i o n

consumed a f t e r

1. 58 ml.

of 0.0103

N sodium h y d r o x i d e ) .
1.6 e q u iv a le n t s
hydrolysis

for

five

of 0 ,N -acety l
hours

) (5-lj- rag. c o n s u m e d a f t e r

and d i s t i l l a t i o n

1.63 ml.

of 0.0103

N sodium h y d r o x i d e ) .

“*) The c a l c u l a t i o n s
s ine .

are based

on a d i a c e t y 1 so 1 a n o c a p -

100

CO

CO

o

o

OJ

uoj ss j uistre jx T-uao Jaj
Infrared

Spectrum

of

0,N

- Diacetylsolanocapsine

CO

11.

in Mi c r o n s

o

Fi gur e

Wavel engt h

( Nuj ol

M ull).

83

oj

100

Spectrum

of

o
oo

‘O

uor ss

tuisub jx

m ao Jad

o
0,N,N* - T r i a c e t y l s o l a n o c a p s i n e

CO

Infrared

o

12.

in M i c r o n s

o

Fi gure

Wa v e l e n gt h

(Nuj ol

Mull).

81*

'L-T\

85
0.9

equivalents

hydrolysis

of 0 - a c e ty l* )

and t i t r a t i o n

0.91 ml.

( 5 . 7 mg.

consumed a f t e r

of 0. 0103 N sodium h y d r o x ­

ide ) .
D i a c e t y l s o 1a n o c a p s i n e wa s

soluble

A solution

t h e compound/ml.

of

0 . 2 0 mg.

s h owe d no a b s o r p t i o n
b.

ultraviolet

solanocapsine
in an o i l

( 6 ) wa s f o l l o w e d .

and

10 m l .

bath for

diminished p ressure

five

of

product melting

a nd t h e

brown,

to

at

did not

melting point

of

150-160°C.
A further

material

into

on a l u m i n a .
in 5 m l . of

it

spectrum.

o f 5 0 0 mg.

turbidity

Thus a y i e l d

solution

of

dissolved

in

a n d t h e c om­

o f J4.OO mg.

of

The c ompound

in h o t

a crystalline

the p u r i f ie d

benzene-

c o mp o u n d .

c o mpo und wa s 1 5 0 - 1 5 5 ° C .

The
(Lit.

[ 6 ]).

a t t e m p t wa s ma de t o c o n v e r t
a crystalline

A solution

the

amorphous

c o mp o u n d by c o l u m n c h r o m a t o g r a p h y

o f 8 3 mg.

of a c e t y l a t e d

b e n ze n e was c h r o m a t o g r a p h e d

“ ) The c a l c u l a t i o n s
s i ne .

to

the

a nd F r a e n k e 1- C o n -

residue

1[]_0-150°C. wa s o b t a i n e d .

change

of methanol

s o l v e n t wa s r e mo v e d u n d e r
oily

slight

crystallize.

acid.

a n h y d r i d e wa s r e f l u x e d

The

wa s a m o r p h o u s a n d e v e n r e p e a t e d

value

of

A mixture

acetic

hours.

W a t er was added

pound a l l o w e d t o

ligroin

part

Tr i a c e t y l s o l a n o c a p s i n e . - B a r g e r

rat' s procedure

ethanol.

in the

of

in 6 N a c e t i c

are based

on 9 g.

solanocapsine
of

alumina

on a d i a c e t y 1 so 1a n o c a p

86
and e l u a t s
t i o n of

fractions

the

o f 20 m l .

were c o l l e c t e d .

f r a c t i o n s was a s f o l l o w s

The c o m p o s i ­

in Table XVIII.

TABLE XV 111
FRACTIONATION OF TRI ACETYL SOLANOCAPSINE BY COLUMN
CHROMATOGRAPHY ON ALUMINA
F r a c t ion

E 1u a n t

Re s idue

1-3

benzene

k-5

benzene +

5% c h l o r o f o r m

k mg. c o l o r l e s s o i l ,
s o l u b l e in e t h e r

6-7

b e n z e n e + 10% c h l o r o f o r m

6 mg . c o l o r l e s s o i l ,
s o l u b l e in e t h e r

8-9

b e n z e n e + 29% c h l o r o f o r m

k mg. c o l o r l e s s o i l ,
s o l u b l e in e t h e r

10-13

c h l o r o f o r m + 1% m e t h a n o l

5 6 mg . wh i t e f 0 a m, i n s o l ­
u b l e in e t h e r

111-16

c h l o r o f o r m + 2% m e t h a n o l

5 mg . g r e e n i sh o i l , i n s o l u b l e in e t h e r

Fractions
so 1 v e n t

gave

For
at

17 mm.

5 mg . b r o w n i s h o i l ,
u b l e in e t h y l

1 0 - 1 3 w e r e c o m b i n e d and a f t e r

an amorphous p r o d u c t

analysis
pressure

the

expected

still

triacetyl

e v a p o r a t i o n of

melting

at

derivative

over phosphorus p e n to x id e

Found:

C 9 71-17;

H,

C,

H, 9i| 9 %

71-61;

l50-l55°G.
wa s d r i e d

and p a r a f f i n

r oom t e m p e r a t u r e .
A n a l . C a l c f d f o r C^ 3H^ 2 N2 ° 5 :

sol­
ether

9l| 2 %

at

87
D eterm in atio n of e q u iv a le n t s
equivalents

2 .8

hydrolysis

for

eight

of

of 0 ,N -acety i
hours

acetyl

groups:

) ( i | . 3 mg.

and d i s t i l l a t i o n

consumed a f t e r
ml.

2 .11

of 0 . 0 1 0 3

N sodium h y d r o x i d e ) .
Triacetylsolanocapsine

wa s f o u n d

A solution

of

insoluble

in 6 N a c e t i c

ac i d .
of

only endabsorption
11 .
a.

0 . 2 0 mg.
in the

ultraviolet

part

(150 mg.)

of S c h l i t t l e r
was r e f l u x e d

b a t h w i t h 0 . 7 n l . o f 1|0 p e r
cent

formic

acid.

cent

for

four

formaldehyde

E v o l u t i o n of gas

oil.

5 m l . of w ater

and 2 m l . o f N h y d r o c h l o r i c

s o l u t i o n was f i r s t

and e v a p o r a t e d

and w e i g h e d
anol,

h o u r s on a w a t e r
and 0 . 8 m l .

soon began

extracted

four

acid.

times with

The

10 m l .

ammoniacal w i t h d i l u t e

1 IpO mg.

of

in t he

combined,

dried

to dryness.

over

R e c r y s t a 11 i z e d t w i c e

t h e c o mp o u n d m e l t e d

" ) The c a l c u l a t i o n

at

anhydrous

The r e s i d u e

208°C.

(Lit.

acidic

of e t h y l

ammoni um

and e x h a u s t i v e l y e x t r a c t e d w i t h e t h e r .

e x t r a c t s were

(78).

The r e a c t i o n p r o d u c t wa s d i l u t e d w i t h

t h e n ma de d i s t i n c t

hydroxide

fate

spectrum.

and U e h l i n g e r

yellow colored

ether

of the

Miscellaneous Derivatives

the procedure

Solanocapsine

ether,

m e t h a n o l s howed

Tr i m e t h y l s o 1a n o c a p s i n e . - The d e r i v a t i v e wa s p r e p a r e d

following

100 p e r

t he compound/ml.

The

latter

sodium s u l ­

wa s c r y s t a l l i n e

from a b so lu te meth­
v a l u e 2 09 C.

[78]).

i s b a s e d on a t r i a c e t y l s o l a n o c a p s i n e .

85

For
dried

analysis

the

expected

t r i m e t h y l s o 1a n o c a p s i n e wa s

over phosphorus pentoxide

and p a r a f f i n

at

17 mm. p r e s ­

sure .
A n a l . C a l c 1d f o r

c 2 qh ^

2 N2 ° 2 :

^
N,

b.

So 1 a n o c a p s i n e

5 8 6 , 572 % ( K j e l d a h l )

p i c r a t e . - The p i c r a t e

described by S c h l i t t l e r

a nd U e h l i n g e r

(100 mg.)

in 3 ml.

of

was d i s s o l v e d

saturated

ethanolic

s o l u t i o n was h e a t e d
w a t e r wa s a d d e d .
After

standing

picric

to

19 9 ° C.

(Lit.

c.

I4.O mg.

value

dropwise w ith
When t h e

calculated

cent

a copious p r e c i p i t a t e
washed w i t h
the

absolute

derivative.

228- 289°C.

d.

needles

crystallized.

166 mg.

of c ru de

recrystallized

a melting point

of

of

198-

absolute
acid

e t h a n o l wa s r e a c t e d

in a b s o l u t e

The d e r i v a t i v e

and gave

of

A s o l u t i o n of

a mo u n t o f e x a l i c

appeared.
ethanol

from

[78]).

oxalic

equimolar

The

of b o i l i n g

( 7 8 ) wa s f o l l o w e d .

in 3 ml.

10 p e r

gave

excess

was a d d e d .

o x a l a t e . - Again the procedure

a n d Ue h l i n g e r

so 1 a n o c a p s i n e

fine

The d e r i v a t i v e ,

200-20i°C.

A small

a nd 5 m l .

refrigerator,

( 1 : 1 ) and d r i e d ,

So 1a n o c a p s i n e

Schlittler

point

as

So 1a n o c a p s i n e

(1.25 ml.)

By s l o w l y c o o l i n g ,

p i c r a t e was o b t a i n e d .
e t h a n o 1- w a t e r

acid

in the

(78).

of e t h a n o l .

the b o i li n g

12 h o u r s

wa s p r e p a r e d

after

ethanol.

a c i d wa s a d d e d ,
wa s t h o r o u g h l y

d r y i n g l±2 mg.

I t s m e l t i n g p o i n t wa s 2 8 7 - 2 8 9 ° C .

(Lit.

of

value

[ 78]).

1 s o p r o p y 1 i d e n e s o l a n o c ap s i n e . - The c ompou nd wa s made

following

the procedure

of Barger

a nd F r a e n k e 1 - C o n r a t

( 6 ).

100

39

90

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3
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cd
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o

01

a,
O
o
o
c
O

o

O
O
CO

•— I

2*">
x
-t—I
CD

t-.
H

Oh
o
e
+->

O
Cl)
O h

T-T\

CO
*o
i~
(0
CD

to
C
O
O
O
•tH

u
Oh
O
'—l
-

c

«—H

•*-H
<d

t-

+->
CD
C

*r~t

Cl)

u .

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100

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o

C\J

o

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u oj ss^uisub j x

dad

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CD

91
So 1a n o c a p s i n e

(70mg.)

water

bath

for

After

standing

and

a few m i n u t e s
for

eight

t a l s were

separated

product.

Recrystallized

pound m e l t e d
The

at

a c e t o n e wa s h e a t e d
crystals

in th e

from e t h y l
(Lit.

began to

refrigerator,
a nd y i e l d e d

appear.
the c r y s ­

67 mg.

acetate-acetone,

v a l u e 233°C.

on a

of

t h e com­

[6]).

t h e c o m p l e x wa s e x a m i n e d b y s h a k i n g t h e

of

or w i t h h y d r o c h l o r i c

acid

and t h e n t e s t i n g

acetone:
1) T e n m i l l i g r a m s

of w a ter

micropipet.
2)

of

in a c e n t r i f u g e

s i o n was c e n t r i f u g e d ,

ml.

until

hours

232-233°C.

stability

of

by c e n t r i f u g a t i o n

c o mp o u n d w i t h w a t e r
for

10 m l .

The

of

of 2 N h y d r o c h l o r i c

a d d e d a nd t h e
tested for

gave

The

for

so 1 a n o c a p s i n e

100 mg.

of p -to lu e n e

in 2 .

s t i r r e d w i t h 0.1

sodium h y d r o x i d e
The

a

iodoform t e s t .

a few m i n u t e s .

of

Three-tenths
s o l u t i o n wa s

s u p e r n a t a n t wa s

solanocaps in e . - A solution

ml ■

sulfonyl

(1:1).

suspen­

i o d o f o r m t e s t wa s p o s i t i v e .

was w a s h e d w i t h muc h w a t e r .

of

p y r i d i n e wa s t r e a t e d w i t h

chloride.

After

standing

resulting

12

precipitate

The p r o d u c t wa s r e c r y s t a l l i z e d
O n l y s o l a n o c a p s i n e wa s r e c e i v e d

e v id e n c e d by mixed m e l t i n g p o i n t
compound.

a negative

o f w a t e r wa s a d d e d and t h e

from e t h a n o 1- w a t e r

The

drawn o f f w i t h

c o mp o u n d w a s

cent

tosylation

100 mg.

s t i r r e d w i t h 0. 1 ml.

a few m i n u t e s .

supernatant

the

acid

10 p e r

acetone.

5> m l .

for

c o n t e n t was c e n t r i f u g e d .

e. A ttempted

hours,

tube

supernatant

of

c o mp o u n d wa s

a nd t h e

Ten m i l l i g r a m s

of a m i l l i l i t e r

the

a nd

infrared

sp ect rum of

as
the

92
f • Attempted
By a c i d

d e h y d r a t ion of

c a t a l y s i s . - A s o l u t i o n of

i n 15 ml . c o n c e n t r a t e d
wa s r e f 1 u x e d f o r
the

sol a n o c a p s ine . -

h y d r o c h l o r i c a c i d a n d 35 ml . s t h a n o l

10 h o u r s .

After

s o l u t i o n wa s ma d e d i s t i n c t l y

four

t ime s w i t h

ether

1 0 0 —m l .

f r a c t i o n s were

evaporated

dried

(1:1).

over

50 m l . o f w a t e r wa s a d d e d ,
ammoniacal

portions

to dryness.

e t h a n o 1- w a t e r

50 m g . so 1a n o c a p s i n s

of e t h y l
anhydrous

and t h e n e x t r a c t e d

ether.

The c omb i ne d

sodium s u l f a t e

The r e s i d u e wa s r e c r y s t a l l i z e d
O n l y so 1a n o c a p s i n e wa s r e c o v e r e d

e v i d e n c e d by m i x e d m e l t i n g

point

and

infrared

a nd

from
as

spectrum of

the compound.
By b a s e
procedure

of Barger

A solution
of

10 p e r

1 0 0 ° C.

c a t a l y s i s . - Dehydration

of

a nd F r a e n k e l - C o n r a t

60 mg.

of

with ethyl

four

hours.

ether,

crystallized
melting point

and

The y i e l d

infrared

( 6 ) wa s a s f o l l o w s :

dihydrochloride

in 5 ml.

which

and F r a e n k e 1- C o n r a t

o x i d a t i o n was p e r f o r m e d

of

solvent,

at

extraction
an amor phous
Re­

( 1: 1) t h e p r o d u c t wa s f o u n d by

s p e c t r u m t o be

is c o n t r a r y

Oppenauer

of w ater,

o f p r o d u c t wa s 36 mg.

so 1a n o c a p s i n e

12.
a.

and removal

f r o m e t h a n o 1- w a t e r

recovered

with acetone,

Up on a d d i t i o n

drying

r e s i d u e wa s o b t a i n e d .

by B a r g e r

so 1a n o c a p s i n e

s o l u t i o n by t h e

c e n t m e t h a n o l i c p o t a s s i u m h y d r o x i d e was h e a t e d

for

The

in b a s i c

so 1a n o c a p s i n e .

gave a c o n d e n s a t i o n p r o d u c t
to

t h e c o mp o u n d t h u s

obtained

(6).

Oxidation Studies
oxidation
following

o f n i t r o s o so 1a n o c a p s i n e . - The
the procedure

of P r e l o g

and

93
Szpilfogel

(71).

N i t r o 30 so 1 a n o c a p s i n e

on s t e a m b a t h w i t h 8 m l .
lute benzene,

and

1 g,

of anhydrous a ce to n e ,

of

20 m i .

tube

containing

The

anhydrous

sulfate.

The r e a c t i o n m i x t u r e w a s r e a c t e d w i t h w a t e r
hydroxide
ethyl

of a b s o ­

2l\. h o u r s .

alumi num phe nox ide f o r

c o n d e n s e r wa s e q u i p p e d w i t h d r y i n g
calcium

( 1 0 0 m g . ) wa s r e f l u x e d

and e x t r a c t e d

ether.

tallized

The y i e l d

three

For

c r y s ta ls melting
analysis

the

( Lp: 1 ) t h e
at

was h e a t e d

solid

of

Recrys­

transformed

to

258-260°.

oxidized

over phosphorus p e n to x i d e
drying p is to l

30-ml. portions

o f y e l l o w r e s i d u e wa s 60 mg.

f r o m e t h a n o 1- w a t e r

colorless

times with

and 6 N s o d i u m

n i t r o s o so I a n o c a p s i n e wa s d r i e d

and p a r a f f i n
by r e f l u x i n g

at

17 mm. p r e s s u r e .

methanol.

Anal . C a l c ’ d f o r C ^ H ^ O ^ :

C,

73.60:

H,

9.15;

N,

6.36 %

Found:

C,

72.89;

H,

9.16;

N,

6.03%

A solution
endabsorption

of

0 . 2 0 mg.

in th e

compound/ml.

ultraviolet

The c o mp o u n d d i d n o t

give

part

The

methanol

of the

showed o n l y

spectrum.

any p r e c i p i t a t e

with d ig ito n in

c o n t r a r y t o n i t r o s o so 1 a n o c a p s i n e ,
b . Chromic

acid

o x i d a t i o n of t r i a c e t y l

The o x i d a t i o n w a s p e r f o r m e d
d a t i o n of
600 mg.
acetic
acid

of

tomatidine
triacetyl

after

by Sato et

the procedure

a l . (76).

so 1a n o c a p s i d i n e

a c i d wa s a d d e d d r o p w i s e w i t h

i n 10 m l .

t u r e was k e p t

of 80 per
at

cent

c a . 1 0 ° C.

so 1 a n o c a p s i n e . -

stirring

by c o o l i n g

acid.
in

oxi­

To a s o l u t i o n o f

i n 20 m l .

acetic

used for

of

glacial

5 0 0 mg.

of chromic

The r e a c t i o n m i x ­
ice-water.

After

100

15.

to

CO

o

o
o
vO
uoj ss juisueux T-'U^D d3d

o
D e r i v a t i v e Obt a i n e d
(Nujol M u l l ) .

00

10
c
o
u
(J

A
+->
CD
c
0)
«—
I
(0
>
cd
:s

I n f r a r e d Spectrum of
Nitrososolanocapsine

after

o

Fi gur e

Oppenauer

Oxidation

of

9k

95

standing

at

room t e m p e r a t u r e

s o l u t i o n was p o u re d
The e t h e r e a l
carbonate

1.5 hours,

ice-water

sodium s u l f a t e

and w a t e r

135 rng.

To p u r i f y

cedure

c o mp o u n d f u r t h e r ,

impurities

recrystallization
the

point

70 mg.

( T)

(79)

wa s 61 mg.
of

derivative

semi-crystalline
the

to

of

t h e c ompou nd

separate

the k e t o n i c

the

The p r o ­

wa s f o l l o w e d .

a nd o f

Y i e l d of

"ketonic" material

"ketonic" material
158-160°C.

(0.20 mg./ml.

wa s a f t e r

The u l t r a v i o l e t

methanol)

showed a n

2 8 5 mp.

elementary

temperature

of

anhydrous

non-ketonic m aterial.

from methanol

a b sorption peak at
For

or

by f t e i c h s t e i n

The m e l t i n g

spectrum of

over

sodium

R e c r y s t a l 1 i.zed f r o m d i l u t e m e t h a n o l

"non-ketonic'1 m aterial
10 mg.

dilute

1 3 i ^ - 1 3 5 ° C.

the

described

dried

The y i e l d

wa s t r e a t e d w i t h G i r a r d 1 s r e a g e n t
from

dark-brown

and e x t r a c t e d w i t h e t h e r .

again,

and e v a p o r a t e d .

m e l t i n g p o i n t was

material

the

s o l u t i o n wa s w a s h e d w i t h w a t e r ,

solution

p r o d u c t was

into

for

analysis

the

derivative

over phosphorus pentoxide

wa s d r i e d

and p a r a f f i n

at

a t r oo m
17 mm.

pressure.
An a l .

Found:

C,

Lj.2.82; H,

8.80;

It has not been p o s s ib l e
a reasonable

empirical

The d e r i v a t i v e

dinitrophenylhydrazine

13.06 %

from these

formula for

gave

N,

analyses

the o x id a tio n product.

a yellow p r e c ip ita te

(79).

to c o n stru ct

with 2 , 6 “

16.

U ltraviolet

of

OJ

o

o

OJ

o

CO
OJ

o

o

o

o
A oueqjosqv

o

o
Oxidation

o
o

Spectrum

Product

of

T riacetylsolanocapsine.

CO

Fi g ur e

96

97

001
h o t

s s juisine j x ^ u a o

jo j

13.

Color R eactio n s

liebermann-Burchard

color

test

as d e s c r i b e d by F i e s e r (28)

wa s n e g a t i v e

with

ocapsine

Mn i t r o s od i h y d r o so 1 a n o c a p s i n e . TT

and

so 1a n o c a p s i n e , b u t p o s i t i v e w i t h n i t r o s o l a n -

Rosenheim co lo r

test

(83)

gives

an

immediate

r e d c o l o r w i t h compounds c o n t a i n i n g

a conjugate

system or

-position

group.

a double bond

in the

N i t r o so so 1 a n o c a p s i n e

ga v e no c o l o r

selenium dioxide
to

a double

bond.

of the

group

steriod.

selenium,

test

in a c e t i c

the p o s i t i o n
in t h e

intense

d oubl e bond
a hydroxyl

a nd T,n i t r o s od i h y d r o so 1 a n o c ap s i ne "

reaction.

S e 1e n i um d i o x i d e

adjacent

to

a nd

(28) .
acid

detectable

first

can

The e a s e

d o u b l e bond
The

The m i l d

oxidation

introduce

an a c e t o x y g r o u p

o f o x i d a t i o n d e p e n d s on

in r e l a t i o n

selenium dioxide

to the hydroxy
is reduced to

as a y e l l o w c o l l o i d a l

a nd t h e n a s a r e d p r e c i p i t a t e .

agent

solution

B o t h n i t r o s o so 1a n o c a p s i n e

a n d " n i t r o s o d i h y d r o so 1a n o c a p s i n e ” g a v e a r e d p r e c i p i t a t e
100°C.,

but

not

at

20°C.

o r ]+0°C.

at

IV.

D I S C U S S I O N AND CONCLUSI ONS

I so I a t i o n . - The p u r p o s e
and c h e m i c a l l y

identify

the

of

this

r e s e a r c h wa s t o

antibacterial

isolate

substance(s)

in

S o l anum p s e u d o c a p s i c u m L .
L u c a s and a s s o c i a t e s
cular

reported

a c t i v i t y was a s s o c i a t e d w i t h

a c t i v i t y was p r e s e n t
the

(32)

case,

of th e

since

the

in the

tops.

s u b s t a n c e wa s f o u n d

the

roots

antituber-

and t h a t

no s u c h

T h i s wa s n o t f o u n d t o be

ant itu b e rc u la r

t o p s was a p p r o x i m a t e l y t h e

tubercular

the

that

activity
s a me .

of the

roots

The p r i n c i p a l

t o be t h e

steroidal

and

anti-

alkaloid

so 1a n o c a p s i n e .
Besides,

a substance

p n e u m o n i a e was p r e s e n t .
extract
found
The

of

if

the p lan t

of

identify

ethyl

ether,

b u t no a c t i v i t y was

extracted with ethyl

extract.

to

separate

isolate

ether.

green colored

I t had a c e r t a i n

and w a s v e r y d i f f i c u l t

a f f i n i t y for
from the

a nd c h e m i c a l l y

substance.

So 1a n o c a p s i n e
which h a s not be en
wa s a s s u m e d u n t i l
present

growth of Di p l o c o c c u s

c o u l d be r e m o v e d f r o m a n e t h a n o l i c

No a t t e m p t wa s made t o
this

the

s e e me d t o be c o n n e c t e d w i t h t h e

the p l a n t

so 1 a n o c a p s i n e
alkaloid.

It

t h e p l a n t was d i r e c t l y

substance

fraction

with

inhibiting

is the

only alkaloid

isolated

as a g l y c o s i d e .

c o n tr a r y evidence

in t h e p l a n t

from the

as a g l y c o s i d e .
99

that

genus Solanum

In t h i s

study

so 1a n o c a p s i n e wa s

Either

a glycosidase

it

100
catalyzed
of

the

hydrolysis

harvesting

able

to

or

the

hydrolyze

acid

the

moiety.

fore

by e i t h e r

or

in

ethanol

so 1a n o c a p s i n e .

Fairly

partially

thought

that

tography.
the

but

could

be

One m u s t
artifact,
That

but
a

cussion

of

the

the
have

moiety has
an

acetal

seems

has

nucleus.

so 1a n o c a p s i n e

and

enzyme

a glycoside,

but

amounts

of D-glucose

were

preparations.
in t h e s e

It

and

used

solvent

was

tried

change

the

result;

the

that

plant

water
always
present

was

that
for

for

be­

first

it

chroma­

developing

only

solano-

structural
a primary

All

other

in

this

found

to

be

( 8 i p) .

in

so 1 a n o c a p s i n e

is

not

the

as

isolated.

conclusion will

group

linkage

be

seen

studies,

amino

position

possibility

the

this
for

it

the

dis­

was

found

in p o s i t i o n

alkaloid

and

to

from

where

group

steroidal

attached

The

s a me f o r m

an

3 on

glycosides

carbohydrate

hydroxyl

through

an N - g l y c o s i d e

rem ote.
Barger

nocapsine
ine.

was

in b o i l i n g

solvent

time

procedure

inactivate

plants

the

yield

acidic

not

in

chemical

been

the

conclude

reasonable

a hydroxyi

to

to

at

detected.

so 1a n o c a p s i n e
steroid

isolation

was p re s e n t,

did

glycoside

compound

alkaloid

fairly

present

is

not

reacting
it

the

attempt

large

therefore

it

that

the

A basic

alkaloid,

capsine

did

purified

in

in

placing

a glycoside

was h y d r o l y z e d

used

The

absolute

in th e

an e x i s t i n g

precursor

a carbohydrate
isolation

of

a n d F r a e n k e 1- C o n r a t

they

During

found

the

an

(6)

amorphous

isolation

and

reported

compound

purification

that

called
of

besides

sola-

so 1a n o c a p s i d -

so 1a n o c a p s i n e

101
an a m o r p h o u s ,

resinous material

wa s a l w a y s p o s s i b l e
ethanol

to

change

Schlittler

paper

this

the

material

fact

chromatography,

sidine

to c r y s t a l l i n e

(78) observed the

that

it

so 1 a n o c ap s i ne .

same r e s u l t .

This

o n l y o ne c o mp o u n d wa s o b t a i n e d b y

leads to

is n o n - e x i s t e n t

( op . c i t . ) c a l l e d

but

by r e p e a t e d p r e c i p i t a t i o n from aqueous

and U e h l i n g e r

together with

was o f t e n o b t a i n e d ;

the

conclusion that

and what B a r g e r

solanocapsidine

is

soianocap-

a n d F r a e n k e 1- C o n r a t

impure,

amorphous

solano-

cap s i n e .
It
tains

has been

solanine

isolated
graphy,

stated

and

the

non-existent,
capsine

oidal
to

the

the

name o f

to

by p a p e r

so 1a n o c a p s i n e .

suggested that

so 1a n o c a p s i d i n e .

in l i n e w i t h
For

name o f

the

as

t wo c o mp o u n d s we r e
chromato­
Therefore

c o mp o u n d ” so 1a n o c a p s i d i n e ” a p p a r e n t l y

alkaloids.

let

These

s p ro u ts but did not,

i t m i g h t be

name

S_. p s e u d o c a p s i cum c o n ­

s h o u l d be r e v i s e d .

be c h a n g e d

bring the

(66).

same R ^ - v a l u e

such a s t a t e m e n t
Now t h a t

the p lan t

solanidine

from p o t a t o
give

that

this

the

the
class

aglycone

t h e name f o r

solano-

Such a c h a n g e woul d

nomenclature for
of a l k a l o i d s
end

is

it

other

ster­

is customary

in - id i n e , w h e r e a s t h e

c o r r e s p o n d ing g l y c o s i d e h a s t h e

suffix

- ine.

( 16,14.9,52,57,8!+) .
Concerning

the p o s s i b i l i t y of

tubercular

drug,

it

extremely

large,

but

injected

i n t o mice

c a n be
the

are

said

side

so 1a n o c a p s i n e

that

effects

serious.

the
of

Before

toxicity

as an a n t i is not

t h e c o mp o u n d whe n
one c o u l d e v e n t e s t

102
t h e compound c l i n i c a l l y ,
i n s u c h a wa y t h a t
Chemical
nitrogen

analyses

of

(78)

ization equivalent

weight

for

together with
from the

reagent.

bon gave
digitonin
Ring
after

a value

analyses for
of

C,
the

bromine

o f 1+56, JL+8 1,
a t o m com­
suggested

so 1a n o c a p s i n e

2 7 - c a r b o n atom
A d o u b l e bond

in g l a c i a l

No 0 - m e t h y l
methyl

neutral­

acetic

or N - m e t h y l

groups

groups a tta c h e d to c a r ­

No p r e c i p i t a t e

wa s f o r m e d w i t h

in e t h a n o l .
structure.

- Barger

d e h y d r o g e n a t ion of

Diels hydrocarbon
After

three.

Therefore,

genus Solanum.

acid nor p erm anganate
but

and t h e

a 27-carbon

the other

by n e i t h e r

it

f o r m u l a C2 rj Hl^GN2 ° 2 ’

2 6 - c a r b o n a t o m c ompou nd a s
(6).

and

derivatives,

determination

c o u l d n o t be d e t e c t e d

were p r e s e n t ,

hydrogen,

in a greement w i t h S c h l i -

a g r e e mo r e w i t h

and F r a e n k e 1- C o n r a t

isolated

its

it

disappeared.

so 1 a n o c a p s i ne g a v e v a l u e s

These r e s u l t s

c a n be c l a s s i f i e d
alkaloids

effects

has the em pirical

pound t h a n w i t h a 2 ^ - o r
by Ba rger

and

so 1a n o c a p s i n e ,

Rast' s molecular

and 1+82.

side

so 1 a n o c a p s i n e

that

and Uehl i n g e r

•

undesirable

s t u d i e s . - By m e a n s o f c a r b o n ,

c a n be c o n c l u d e d
tler

the

i t w o u l d be n e c e s s a r y t o m o d i f y

sam e c o m p o u n d .

authentical

TTso 1 a n o c ap s id i n e TT w i t h

of

tube,
Since

so 1a n o c a p s i n e w i t h
Schlittler
this

isolated

selenium,

and u l t r a v i o l e t

selenium at

and W e h l i n g e r

c ompound wa s

c o mp o u n d w i t h r e s p e c t

melting point

(6)

( 3 f - m e t h y l c y c 1o p e n t e n o - p h e n a n t h r e n e ) ( I ) .

12 h o u r s h e a t i n g

in an e v a c u a t e d

a n d F r a e n k e 1- C o n r a t

(78)

identical

to m elting p o in t,

spectrum,

no d o u b t

320

isolated
with the

mixed

remains

103
about

s o l a n o c a p s ine b e i n g

extraction

of

a steroidal

t h e mat e r i a l

obtained

alkaloid.

Through acid

f r om s e 1e n i um d e h y d r o g e n a -

CH 3
2 HC— (\

( I I )

>
/

CH3

CIU
I ^
9 IHG

(III)

CH3
t i o n of

so 1a n o c a p s i n e , t h e

latter

c o mp o u n d 2 - e t h y l ~ ^ ~ m e t h y l p y r i d i n e
Conrat

(6)

isolated

5-m e t h y l p y r i d i n e

and

after

one w i l l

(II).

isolated
Barger

as the

only

and F r a e n k e l -

selenium d e h y d r o g e n a t i o n b o t h 2 - e t h y l -

its

I f one c a n a t t a c h
ships,

authors

assume

i s o m e r l4_“met h y l - 2 - e t h y l p y r i d i ne ( I I I ) .
e.
any importance to biogeiytic r e l a t i o n ­
that

so 1a n o c a p s i n e h a s a s o l a s o d a n
24

CH
-H3

CH

CH

27

(v)

( iv)
structure

( I V) .

Since

w i t h 27 c a r b o n a t o m s

it

is g e n e ra lly thought

are p r e c u r s o r s for

the

that

steroids

steroidal

alkaloids

I ok
(16),

it

does not

that Barger
(III)

seem l i k e l y

relationships

a n d F r a e n k e 1 - C o n r a t * s i p - m e t h y l —2 —e t h y 1p y r i d i n e

was c o r r e c t l y

isolated,

from b i o g e n e t i c

identified.

If

an u n e x p e c t e d p r e c u r s o r

have had a m e t h y l

group a t

this

for

c o mpou nd r e a l l y wa s

so 1a n o c a p s i n e

c a r b o n 2 Ip i n s t e a d

of

at

should
the usual

c a r b o n 25 p o s i t i o n ( I ) .
Functional
a nd S c h l i t t l e r
possesses
hydroxyl

g r o u p s . - Both Barger
and U e h l i n g e r

a p r i m a r y amino,
group.

(78)

and f o r m a l d e h y d e ,

which

found t h a t

the

for

atoms w i t h f o r m i c

a trimethyl

evidence

for

wa s g i v e n b y S c h l i t t l e r
capsine with n itro u s
and

i t wa s f u r t h e r

with 2 e q u iv a le n ts
nitroso
sis
of

derivative,

indicated

that

the e x iste n ce

a nd U e h l i n g e r

acid.

In t h i s

of th e

(78)

at

355

of a n i t r o s a m i n o
( VI )

two a mi n o g r o u p s

study the

solano-

same wa s o b s e r v e d

d e t e r m i n e d t h a t by r e a c t i n g

so 1a n o c a p s i n e

of n i t r o u s

unsaturated

acid,

a neutral,

^27^1p2^2^3, r e s u l t e d .
1 mole

a mi n o g r o u p w a s p r e s e n t .

intensity

investigation.

by r e a c t i n g

A Va n S l y k e

The u l t r a v i o l e t

derivative

(Figure

( I ° 9 E 2.1p2)
group.

analy­

o f n i t r o g e n wa s l i b e r a t e d p e r mo l e

so 1a n o c a p s i n e , f r o m w h i c h c a n be c o n c l u d e d

obtained nitroso

acid

derivative

T h i s o b s e r v a t i o n was c o n f i r med by t h e p r e s e n t ,
Further

a nd a

h a v i n g t wo a mi no g r o u p s

nitrogen

gave

(6)

so 1a n o c a p s i n e

a s e c o n d a r y a mi n o g r o u p ,

The e v i d e n c e

came f r o m m e t h y l a t i o n o f

a nd F r a e n k e 1 - C o n r a t

9)

that

a primary

s p e c t r u m of t he

s howe d a p e a k o f

and s u g g e s t e d

l ow

the presence

D i ( e y e 1o h e x y l m e t h y l ) - N - n i t r o s a m i n e

s h o ws t wo a b s o r p t i o n p e a k s

at

2)p0 mji ( l o g E 3 . 9 )

and

105

c h 2— in

—c h 2

^

N -N -0

( VI )

(vii)

355 inji ( l o g E 1 | . 2 5 ) ,

while

a t 2 5 5 my ( l o g E J+. 25)
peak a t

the

longer

1- n i t r o s o p i p e r id ine

a n d 3 ^ 0 mp ( l o g E 2 . 0 )

w a v e l e n g t h wa s p r e s e n t

(VII)
(38).

has peaks
Only the

in n i t r o s o s o l a n o c a p -

s ine .
I t was assumed by B a r g e r
during the

formation

o f n i t r o s o so 1a n o c a p s i n e

group, o r i g i n a l l y p r e s e n t
water,

in t he m o l e c u l e ,

whereas S c h l i t t l e r

likely that

the

in t h e

leave

group

the hydroxyl

a mi n o g r o u p s

intact.
since

often resu lts

in t h i s

Conrat*s f i n d in g s

that

the hydroxyl

wa s e l i m i n a t e d

(7 8 ) thought

as

i t mor e

a c i d o n t h e p r i m a r y ami no

in the

The l a t t e r

s e e ms l i k e

a

d e a m i n a t i o n of p r i m a r y
f o r m a t i o n of both u n s a t u ­

c o r r e s p o n d i n g h y d r o x y compounds.

study

so 1a n o c a p s i n e , w h i c h

(6)

f o r m a t i o n o f a d o u b l e b o n d and

explanation,

r a t e d compounds and t h e
experiences

and U e h l i n g e r

a c t i o n of n i t r o u s

group would r e s u l t

mo r e p l a u s i b l e

a n d F r a e n k e 1- C o n r a t

i t wa s n o t p o s s i b l e

to dehydrate

is c o n tr a d ic t o r y to Barger

and c o n s e q u e n t l y

supports

Fr om

a nd F r a e n k e l -

Schlittler

and

Uehlinger*s conclusions.
Nevertheless,
of Barger

t h e r e ma y be v a l i d i t y

and F r a e n k e l - C o n r a t

S o 1a n o c a p s i n e

does not

give

in t h e

( o p . c i t . ) due t o

any p r e c i p i t a t e

contentions
the f o llo w in g :

with dig ito n in

and

106
this

i s why t h e

can not

hydroxyl

group

s h o u l d n o t ho 3 ^ —h y d r o x y .

be a 3 c^. —h y d r o x y g r o u p e i t h e r ,

been p o s s i b l e

to dehydrate

gives a p r e c ip ita te
therefore,

that

Digitonin

such a group.

with digitonin,

and

soluble

with

3 c*, - h y d r o x y

that

a r e mu c h mo r e

which

complexes with

steroids.

The

soluble.

latter

Usually,

i t woul d ha ve

N i t r o s o so 1a n o c a p s i n e
i t m u s t be a s s u m e d ,

t h e p r i m a r y a mi n o g r o u p wa s
is a g ly co sid e,

form s p a r i n g l y

since

It

in t h e

3 position.

is g e n e r a l l y regarded
3($ - h y d r o x y ,

to

but not

type pro d u ce s complexes
only 3^ -hydroxy s te r o id s

VH3

CHO
OH
(c h ^

n

OH
( I X)

(VIlI)
( w i t h 3 -OH a n d
alkaloids

10- CH^ c i s t o e a c h o t h e r )

tomatidine,

solanidine,

tated as d ig ito n id e s .

Exceptions

c ompounds a r e
group and
(IX),

conessine

which has

an a l d e h y d e

among n a t u r a l l y o c c u r r i n g

which has a

group

Haworth e t
derivatives

a nd

al .

group

in the

is p r e c i p i t a t e d

(U-1-I-) h a v e

and f o u n d

steroidal

s ol a so d ins are p r e c i p i ­

is p r e c i p i t a t e d with d i g i t o n i n

3 p-hydroxy

tene

(VIII),

and

such as t he

(10)

- d ime t h y 1 am i no
and s t r o p h a n t i d in

10 p o s i t i o n b e s i d e s

with d ig ito n in

synthesized

several

a

(70).

aminocholes-

t h a t / V ^ - c h o 1 e s t e n e - 3 ^3- a m i n e

(X)

107
A5
n
and & - c h o l e s t e n e - 3 p - I s o p r o p y l i d e n e a mi n e
3o( - e p i m e r s ,

readily

Be r t h o et_ aA_.
a mi n e

(XII)

(11)

yielded

reported

(XI),

that

A 7 - c h o 1 e s t e n e - 3 |3 - d ime t h y l -

with d ig itonin.

CHt
I 9
CH-CHo'CHp-CHp'CH-

(XII)

= ( c h 3 ) 2 c =n -

Since
it

iL

R = -NH2
r

(x

so I a n o c a p s i n e

g a v e no p r e c i p i t a t e

c a n now be a s s u m e d b y a n a l o g y w i t h o t h e r

that

the

i i i

)

CH

R = (CH3 ) 2N= c y 1QN-

r

with d igitonin,
3 - a m i n o c o mp o u n d s

a mi n o g r o u p p r o b a b l y h a s a 3 <*.- c o n f i g ur a t i o n .

i n v e r s i o n o f c o n f i g u r a t i o n must have happened d u r i n g
deamination.
as

involving

Ingold
an

( l_j_6) e x p l a i n s

intermediate

inverted
L-RNH
H ills

+
(59)

with n itro u s
reaction.

configuration,

has described
and found

Equatorial

the

and a f f o r d s

i on

an a l c o h o l

of

e.g.:

HNO ------- » RN2 +

acid

An

the deamination re a c tio n

f o r m a t i o n of a d i a z o n i u m

whi ch de compo se s by an S^l p r o c e s s
large

the

p r e c i p i t a t e s with d ig ito n in .

H-CH2'CH2 -CH2'CH-CH

(x i )

not

a n d Z^\7 - c h o l e s t e n e - 3 p - p i p e r id i n e (XI 1 1 ) g a v e no

precipitate

(X)

but

it

> R+

> D-ROH

+

D. L-ROH

d e a m in a t io n of am inodec alins
t o be a t r u e

a mi n o g r o u p s

afforded

s t e r e o spec i f i c
a l c o h o l s of

same

108
conf i g u r a t ion,

while

of c o n f i g u r a t i o n .
the

3

axial

That

-hydroxy group

configuration

a mi n o g r o u p s r e a c t s d

is

in a g re e m e n t w i t h t h e

in s t e r o i d s w i t h the

as c h o l e s t e r o l

t h e r m o d y n a m i c a l l y mo r e

and a b o u t

This

changed
observed

so 1a n o c a p s i n e

t o +J4.O0 .

the

Schlittler
by r e d u c t i o n

l i t h i u m aluminum h y d r i d e .

A Va n S l y k e

dihydro

c o mp o u n d

s howed t h a t

specific

give

n i t r o samine

the

ultraviolet

(Figure

F was o p e n e d a s

r oom

9).

it

illustrated

r o t a t i o n had

a nd Ue hl i n g e r
of

(78)

so 1 a n o c a p s i n e w i t h

a mi n o n i t r o g e n

2 mole s of

g i v e n o f f p e r m o l e o f c o mp o u n d and t h a t
did not

at

s howe d no d e p r e s s i o n o f m e l t i n g

same r e s u l t s

on t h e

the

and t h e r e f o r e ,

gave a d e r i v a t i v e w i t h m e l t i n g p o i n t

so 1 a n o c a p s i n e , b u t

fr om +20°

that

3 atmospheres p r e s s u r e s with reduced

derivative

p o i n t mixed w ith

concept

stable.

platinum oxide c a t a l y s t
2 10-212°C.

inversion

same s t e r e o c h e m i c a l

is e q u a to r ia l

R ing o p e n i n g . - H y d r o g e n a t i o n of
temperature

by

the

analysis

n i t r o g e n wer e

res u ltin g product

absorption c h arac teristic
is th e r e f o re
in t h e

CH

concluded

for

that

a

ring

following:

HN02 C H 3

3

The h y d r o x y

— CH 3

Pt O^

steroid

obtained

s o l anoc ap s i n e w i t h

nitrous

1 mole o f h y d r o g e n

and gave

This hydroxy

steroid

acid

after

r e a c t i o n of d i h y d r o ­

absorbed

a precipitate

derivative

is not

a little

mor e t h a n

with dig ito n in .

completely

identified,

solanocapsine

riNO-

n itrososolanocapsin e

^ —

—Ci

CH

C C )= (C )
dipvdrcsc a n o c a

diq'tom n

d

sine

HNO2

precipitate
H3

ni t r o s o d ' h vd r c t o i a n o c a p s i se n
/

HO— \

r*-u

^

i

<

CH,

(c)=(0
Figure

17.

R e a c t i o n o f So 1a n o c a p s i n e
and D i h y d r o s o 1a n o c a p s i n e
with N itrous Acid.

precipitate

110
since

the

carbon

elementary analysis

i s mo r e

a t o m c o mp o u n d t h a n a 2 7 - c a r b o n

The r e a c t i o n s

of

d i h y d r o s o 1a n o c a p s i n e

nitrous

in agreement wit h
a t o m c o mp o u n d .

acid with

as d e s c r i b e d

a 25-

so 1a n o c a p s i n e

above a r e

and

summarized

in

F i g u r e 17.
Schlittler
spectrum of
intense
free

base.
at

of

band

Besides,
6.1

authors

ride

(78)

observed th a t

t r i m e t h y l s o 1a n o c a p s i n e h y d r o c h l o r i d e

hydroxyl

a band
the

and U e h l i n g e r

the

opened under

than the
the

spectrum of

sp ect rum of

ji c h a r a c t e r i s t i c

of

suggested

that

trimethyl

derivative

influence

of

its

infrared

showed a m o r e

corresponding

the hydrochloride

a C=N b a n d .

r i n g E of the

s howed

To e x p l a i n

in the p r o d u c t i o n of

acid

the

this

the hydrochlofree

base

(XIV)

(XV):

NaOH
(XV)

( XI V)

under

Furthermore,

they proposed

that

influence

a c i d opened

in the

no s i m i l a r

of

evidence

of

this

These o b s e r v a t i o n s
spectra

(Figures

Acetyl
pared

13 a n d

a s c a n be

acetyl

s i d e r e d b y t h e m t o be

so 1a n o c a p s i n e

same m a n n e r , b u t f o u n d

infrared

spectrum.

seen from the

infrared

1]+) h a v e b e e n c o n f i r m e d .

derivatives.

an a mor phous

in the

r i n g E of

- Schlittler
derivative

and U e h l i n g e r
of

(78) p r e ­

so 1a n o c a p s i n e , c o n ­

a N, N - d i a c e t y l so 1a n o c a p s i n e .

Ba^ed

111
up on c a l c u l a t i o n
analysis

gave

for

2 acetyl

11+.97 p e r

groups

cent

of the

pound w h e r e a s

theoretically

t h i s work th e

amorphous d i a c e t y l

tain

11+.22 p e r

rather

cent

acetyl

an 0 , N - d i a c e t y l

it

and

they found
total

s h o u l d be

weight

derivative

acetyl

o f t h e c o m­

16.73 pe r

derivative

suggests

that

cent.

In

wa s f o u n d t o c o n ­

that

the

compound

t h a n a N,Nf - d i a c e t y l

is

deriva-

t i ve .
The r e a s o n s
The d e r i v a t i v e
that

a basic

spectrum of
tio n bands

for

was

such a c o n c l u s i o n are

soluble

in d i l u t e

group p r o b a b l y
the

for

derivative
an O - a c e t y l

group

N-acetyl

(CH^CO-NHR)

Normally the
imately the
found
tive

t o be t h e

of t h e
band;

this

than

it

at

6 . 0 6 p,

given

infrared

for

a s e c o n d a r y a mi d e o r
The s p e c t r u m o f

in F i g u r e

s p e c t r u m of

11.

that

the

is approx­

(8).

T h i s was

diacetyl

same a s t h a t

f o u n d t o be t h e c a s e

a nd s i n c e

one c h a r a c t e r i s t i c
i t ma y b e c o n c l u d e d

is

intensity

of the

for

s h owe d a l i t t l e

6.1+5 p b a n d ,

deriva­

a p r i m a r y a mi n e

a s e c o n d a r y a mi d e t h e
the

absorp­

and 9 . 7 5 p)

i n s t e n s i t y o f an e s t e r

is concluded

was n o t

(CH3CO-NRR* ) o n l y h a s
occurring

the

is u s u a l l y

the

for

1)

indicating

(5»78 p, 8 . 0 0 p,

o f a s e c o n d a r y a mi d e

The 6 . 0 6 p b a n d

anocapsine.
absorption

for

Furthermore,

6.06 p band
but

that

case

and t h e r e f o r e

acetylated.

is

absorption

same a s

acid

2) The

( 6 . 0 6 p a n d 6.1+5 p ) .

so 1a n o c a p s i n e

following:

s ho we d t h e c h a r a c t e r i s t i c

the bands c h a r a c t e r i s t i c

diacetyl

acetic

is p r e s e n t .

and a l s o

the

the

6.1+5 p

diacetylsol-

mor e

intense

a tertiary

a mi d e

a b s o r p t i o n band
that

the

secondary

1 12

a mi n o g r o u p p r e s e n t
Several
exact
just
of

attempts

so 1 a n o c a p s i n e wa s p a r t l y
to prepare

elementary analysis
as the

less

that

in

the

tate,

as

rather

Barger

lation

acetic

i s mor e

is not

a pure

(6)

it

acetylated
for

five

seems t h a t

in agreement w i t h

the

since

hours

a u th o rs based t h e i r

and a c e t y l

groups.

infrared

an O - a c e t y l

Furthermore,
than

NRR1 ) o n l y

the
the

group

group

insoluble

analysis

in d i l u t e

investigation

analysis

the

on

I t wa s c o n s i d e r e d

an e l e m e n t a r y a n a l y s i s whi ch

a triacetate

calcu­

When r e c a l c u l a t e d

a triacetate.

in t h i s

and o b t a i n e d

t o be a

formula.

i t wa s

so 1a n o c a p s i n e

i t wa s c o n s i d e r e d

cedure

The

of

a N,N1- d i a c e ­

On t h e

prepared

gave

is

Bu t t h e

150-160°C.

The a c e t a t e

higher

c o mp o u n d .

a so 1a n o c a p s i n e w i t h 2 7 - c a r b o n a t o m s ,

be a N , N - d i a c e t a t e

for

the procedure

two a u t h o r s ,

with melting point

elementary analysis
But

the conclusion

after

anhydride

on t h e wr ong e m p i r i c a l

b a s i s of

Therefore,

s u g g e s t e d by t h e

an amo r p h o u s a c e t a t e

diacetate.

the

a diacetylsolanocaps ine,

prepared

a n d F r a e n k e 1- C o n r a t

the

gave

an 0 , N - d i a c e t a t e .

by r e f l u x i n g w i t h

of

(78)

for

never

g r o u p s a l w a y s gave a v a l u e

groups.

and U e h l i n g e r

is n o t,

but

basis

acetyl

amorphous d i a c e t a t e

Schlittler
acetate

expected

d e r m i n a t i o n of

t h a n t wo a c e t y l

a diacetate

acetylated.

is

also

after

to

acids.

their

pro­

in agreement w i t h
indicated

3 acetyl

s p e c t r u m s howed t h e b a n d s c h a r a c t e r i s t i c
a n d a s e c o n d a r y a mi d e

intensity

of t h e

6.1+5 p b a n d .

( CH^ CO- NHR) .

6 . 0 6 p b a n d wa s c o n s i d e r a b l y

Since

s h o ws o n e c h a r a c t e r i s t i c

a tertiary

a mi d e

a b s o r p t i o n band

( CH^CO-

at

6.06 p,

113
it

is concluded

the

that

s e c o n d a r y ami no

considered

or

group.

acetyl

hydroxyl

but not

that

to

is t h e r e f o r e

cholesterol

considered

group.

t o be a t e r ­

T h i s c o n c l u s i o n wa s

This

i n v e s t i g a t i o n has

the hydroxyl

shown

g r o u p c a n be a c e t y -

group. - Nitrososolanocapsine

a positive

quirements necessary

to

except

bond

that

and F r a e n k e 1- C o n r a t

tosylated.

L o c a t i o n of hydroxyl

hydroxyl

derivative

(78)

derivatives.

as p r e v i o u s l y d i s c u s s e d

like

g r o u p wa s a t t a c h e d

The a c e t y l

and U e h l i n g e r

an u n r e a c t i v e

b a s e d on t h e

lated,

acetyl

g r o u p wa s b y b o t h B a r g e r

and S c h l i t t i e r

tiary

third

t o be 0 , N , n ' - t r i a c e t y l s o l a n o c a p s i n e .

The h y d r o x y l
(6)

the

a double

Lieberman-Burchard

give

a positive

test.

reaction

gives

The r e ­

a r e n o t known,

s e e ms n e c e s s a r y t o g e t h e r w i t h

a

group.

Lathosterol

( XVI )

i s o x i d i z e d by s e l e n i u m d i o x i d e

20 ° C . , w h e r e a s c h o l e s t e r o l

( XV I I )

requires

6 0 ° C.

(28)

at
for

ox i d a t i o n .

HO'

HO'

( XVI )

( XVI I )

N i t r o s o s o 1a n o c a p s i n e w h i c h wa s f o u n d t o c o n t a i n a h y d r o x y l
group

(presumably at

the

3 position)

o x i d i z e d by s e l e n i u m d i o x i d e
cluded

that

in e i t h e r

of

the
the

double

at

a nd a d o u b l e b o n d

100 C.

Therefore,

it

io f i r o t
io c o n ­

bond d o e s n o t o c c u p y a p o s i t i o n ao f o u n d

t wo p r e v i o u s l y m e n t i o n e d

sterolo.

The

Rosenheim t e s t
double

bond

s y s t e m or

the h y d r o x y l
to t h i s

is c h a r a c t e r i s t i c

group.

test,

it

for

a d o u b l e bond
Since

suggests

either

a conjugated

in t h e

cX, (3 - p o s i t i o n

n i t r o s o so 1a n o c a p 3 i ne
that

the

d o u b l e bond

to

is n e g a t iv e
is not between

c a r b o n s if a n d 5 .
An O p p e n a u e r
oxidize
If

the

the

o x i d a t i o n of n i t r o s o s o l a n o c a p s i n e

hydroxyl

group

in t h e

3 position

d o u b l e bond p r e s e n t

in t h e

c omp oun d

c a r b o n s If a n d 5 ( X V I I I )
saturated

ketone

to a keto

group.

is between e i t h e r

o r c a r b o n s 5 and 6 ( XI X)

s h o u l d be o b t a i n e d .

( XX)

should

an

<X, (3 - u n ­

S u c h a compound

Oppenauer

XVIII)

Ox i d a t i o n

Oppenauer
(XI X)

Ox i d a t i o n

should
not

show u l t r a v i o l e t

found

t o be

the

double bond c o u l d
It has

case,

been observed

c a n be o x i d i z e d

has not

as yet

the

capsine

contained

it

T h i s wa s

is concluded t h a t

the

to

t h a t m o s t c o mp o u n d s w i t h a s o l a s o d a n
a pregnenolone

b e e n accompl i s h e d

chromic

wa s n o t

hence

a b o u t 2If0 m^i.

n o t o c c u p y any of t h e m e n t i o ne d p o s i t i o n s .

nucleus

since

absorption

acid

oxidation product

completely

a carbonyl

in t h e

identified.

group as

indicated

derivative.

case

of

This

so 1a n o c a p s i n e ,

o f t r i a c e t y l s o 1a n o Nevertheless,

it

by a p r e c i p i t a t e

115
w i t h 2 , l | - d i n i t r o p h e n y l h y d r a z i n e , and t h e
s howe d a b s o r p t i o n

at

2 8 5 mp.

T h i s m i g h t be

double bonds

in c o n j u g a t i o n w i t h

is the

one c o u l d

case,

ultraviolet

imagine

a carbonyl

that before

spectrum

indicative

of

group.

that

If

t wo

d e h y d r a t io n the

Chk
CH3 9 = 0

2
(XXI}
'hydroxyl

g r o u p was a t t a c h e d

(XXI I)

to carbon

1Ip i n a h y p o t h e t i c a l

am i n o p r e g n e n o l o n e

(XXI).

-ol-20-one

s h o ws a b s o r p t i o n a t b o t h 239 a n d 3 0 6 mp

(28).

( XX I I )

This p o s s i b i l i t y

working h y p o t h e s i s

it

attached

to c arb o n 8.

ceivable

that

However,

is t h e r e f o r e

is

then

Placing
line with

the

these

so 1a n o c a p s i n e
yield

Not two,

but

hydroxyl

group

it

is

is con­

c o u l d be / \ ® * ^
derivative

wo u l d

in t he 8 p o s i t i o n w i l l

selenium for

be

in

and U e h l i n g e r :

If

75 m i n u t e s a s m a l l

c o mp o u n d C ^ y H ^ N O wa s o b t a i n e d .

T h i s com­

a p r i m a r y a mi n o g r o u p a nd a h y d r o x y l

three

An u l t r a v i o l e t

this

by S c h l i t t l e r

wa s h e a t e d w i t h

lost

hypothesis

group

2 8 0 - 2 9 0 myi.

observations

of a b a s ic

pound had

and t h a t

As a new

a hydroxyl

oxidation product

a l logregnene-20-one-3-amine
about

this

a l 1o p r e g n e n e - 3

excluded.

suggested

Following

a possible

s how a b s o r p t i o n

1

double

spectrum

bond

s e e me d t o h a v e b e e n

indicated

conjugated

double

group.
introduced
bonds

116
max 2 8 7 muv}.

b ince

p r i m a r y ami no g r o u p
XXIII f o r

the

is

it

.
is proposed

in the

in t h i s wor k t h a t

3 position,

dehydrogenation product

one c a n w r i t e

obtained

the
formula

by S c h l i t t l e r

and U e h 1 i n g e r .
H

(XXIII)

As a f i n a l
(XXIV)
that

is proposed

for

is

t h e n be

this

study the follow ing formula

s o la n o c a p s ine,

n o t v e r y much e v i d e n c e

position
will

c o n c l u s i o n of

submitted.

The

for

although

a hydroxyl

systematic

it

group

name f o r

is r e a l i z e d
in t h e 8

so 1a n o c a p s i n e

3 o< - am i no so 1 a s o d a n - 8 - o 1 m o n o h y d r a t e .
H

(XXIV)
Concerning
succeed
tive.

in o x i d i z i n g
If

such a type

potentially
tisone

future

abundant

and o t h e r

work

i t w o u l d be v e r y d e s i r a b l e

so 1 a n o c a p s i n e

to

a pregnenolone

o f c o mp o u n d c a n be p r e p a r e d ,
raw m a t e r i a l

steroid

for

the

to

deriva­

another

s y n t h e s i s of c o r ­

h o r m o n e s w o u l d be a s s u r e d .

V.

SUMMARY

1 . The a n t i t ube r c u 1 a r p r i n c i p l e
p s e u d o c ap s c um L .
which

is found

in a c o n c e n t r a t i o n

b ac t e r i um t ub e r c u l o s i s .
the p e r i t o n e a l
kg.

by s u b c u t a n e o u s

injection

tested

for

terial

organisms.

2.

In a d d i t i o n

hibiting

b o t h compounds.
against

so 1a n o c a p s i n e

the

in t h e p l a n t

as

1|_. M o d i f i c a t i o n s
i ng a n d p u r i f y i n g

the

a free

alkaloid

alkaloid

are

c o l u m n c h r o m a t o g r a p h y on a l u m i n a ,
wa s n o t
5.
to

d o s e s a r e 200
The a l k a l o i d wa s

as a g l y c o s i d e have
about other

alka­

so 1a n o c a p s i n e

and n o t as a g l y c o s i d e .

of p r o c e d u r e fo r

described.
acid

in­

is p r e s e n t .

is r e g a rd ed t h a t

and v a r i a t i o n s

50-100 mg./

a colored p rinciple

alkaloid

it

into

21 d i f f e r e n t b a c ­

C o n trary to f i n d i n g s

genus Solanum,

injected

200 mg. w h e r e a s

t h e mi ni mum l e t h a l

isolate

successful.

lo id s from the
occurs

to

dose

hydrochi oride

activity

g r o w t h o f Myc o-

solanocapsine

g r o w t h o f D i p l o c c u s p n e umon i a e

3. A t t e m p t s
not been

to

its

for

antibacterial

is fo r

so 1 a n o c ap s i ne ,

inhibits

The m i n i mum l e t h a l

and f o r

of body weight

alkaloid

If'/mL .

of 3

c a v i t y of mice

of body w e ig h t

mg./kg.

t o be t h e

i n t h e p i a n t S o l a n urn

alumina,

extract­

P u r i f i c a t i o n by
or Dowex 50

successful.
It

so 1a n o c a p s i n e

so 1 a n o c a p s i d i n e

has been
contains
(6).

No

stated

that

solanine

S. p s e u d o c a p s i c u m

(66),

solanidine

i n d i c a t i o n of th e p r e s e n c e
U7

in a d d i t i o n

(66),

and

of thes e

118
additional

compounds h a s b ee n f o u n d .

different

solvents

revealed

In r e a l i t y

changed

cent

to

impure,

the

6.

The

in

(dry weight

2 . 214. p e r

solanocapsine

cent

by B a rg er

amorphous

crystalline

chromatography

o n l y o ne a l k a l o i d ,

S o l a n o c ap s i d i n e , a s r e p o r t e d
is

Paper

in

so 1 a n o c a p s i n e .

and F r a e n k e 1- C o n r a t

(6),

s o l a n o c a p s i n e , w h i c h c a n be

form.
content varied

basis)

leaves plus

from 1.53 to 2 .1 5 per

s t e m s a nd f r o m 0 . 8 3

in r o o t s

of p l a n t s

g r o wn d u r i n g

a three

alkaloidal

glycoside

solanine

its

to

year

p e r iod .
7.

The

and

f r o m So 1a num t u b e r o s u m t o g e t h e r w i t h a c r u d e
f r o m S o l a n u m c a r o 1 i n e n s e we r e
s h o we d w e a k a n t i t u b e r c u l a r
glycoside
found to
8.
ical

tomatine
be

It

isolated.

activity.

Besides,

inactive

a g a i n s t M.

has been v e r i f i e d

only solanine

the

alkaloidal
and

tuberculosis.

that

so 1a n o c a p s i n e h a s

the empir­

Schlittler

and

(78 ) .

have

configuration
10.

fraction

f r o m L y c o p e r s i c o n e s c u l e n t u m wa s t e s t e d

9. Assuming a s o l a s o d a n
found to

alkaloid

Of t h e s e

f o r m u l a c 2 7HIj_6N2 0 2 *H2 ° a3 P r o P o s e d

Ueh1 inger

aglycone

a p r i m a r y a mi n o g r o u p
is probably

Schlittler

so 1a n o c a p s i n e

structure,

(78)

so la n o c a p sin e has been

in t h e

3 position.

The

3 (X. .

and U e h l i n g e r ' s
is found

a m o r p h o u s N , N 1~ d i a c e t y l -

t o b e mo r e

identifiable

as 0,N-

diacetylsolanocapsine.
11. E v i d e n c e
solanocapsine

(6)

for

Barger

being

a n d F r a e n k e 1 - C o n r a t ' s N, N - d i a c e t y l -

0 , N, N1- t r i a c e t y l s o l a n o c a p s i n e

io p r e s e n t e d .

119
12. O p e n i n g of
structure
13.
ally

the p i p e r i d i n e

has been accomplished
The h y d r o x y l

assigned
l i f . The

solanocapsine

group

ring

in t h e

solasodan

b y h y d r o g e n o 1y s i s .

i n s o l a n o c a p s i n e may be p r o v i s i o n ­

to carbon 8 .
tentative
is

3

systematic

chemical

name p r o p o s e d f o r

- a m i n o s o 1 a s o d a n - 8 - ol m o n o h y d r a t e .

B IBL IOGRAPHY
1. A l i c i n o , J . F.
Microdetermination
A n a l . Chem. , 20_, 5 9 0 ( 191^8) .

of O - a c e t y l

Groups,

2 . A n g e l 1, H. R . , W a l k e r , J . C . , a nd L i n k , K. P .
The R e l a ­
t i o n o f P r o t o c a t e c h u ic A c i d t o D i s e a s e R e s i s t a n c e in t h e
O n i o n , P h y t o p a t h o l o g y , 2 0 _, I4O I - I 538 ( 1 9 3 0 ) .
3. A r h e r , A.
H e r b a l s , T h e i r O r i g i n and E v o l u t i o n . A C h a p t e r
i n t h e H i s t o r y o f B o t a n y 114.70 —1 6 7 0 , C a m b r i d g e U n i v e r s i t y
P r e s s , 193b.
1|. A z a r o w i c z , E. N . , H u g h e s , I . E . , a n d P e r k i n s , C. L.
A n t i b i o t i c s in P l a n t s of S o u t h e r n C a l i f o r n i a a c t i v e
a g a i n s t M y c a b a c t e r rum t u b e r c u l o s i s 607 a n d A s p e r g i 11 us
n i ge r , A n t i b i o t i c s & C h e m o t h e r a p y , 2 , 532-53"S ( 1 952 )
5.

B a i l e y , L . H.
S ta ndard C yclo p e d ia of H o r t i c u l t u r e , v o l.
I l l , The M a c m i l l a n C o m p an y , New Y o r k , 191-1-7, P~^ 318 3.

6 . B a r g e r , G . , a n d F r a e n k e 1 - C o n r a t , H. L .
A l k a l o i d s from
S o l anum P s e u d o c a p s i c um L . , f . Chem S o c . , 1 9 3 6 , 1537-151+2.

7.

B e l c h e r , R . , a n d G o d b e r t , A. L.
Semi-micro Q u a n t i t a t i v e
O r g a n i c A n a l y s i s , L o n g m a n s , G r e e n a n d C o m p an y , L o n d o n ,
191t 5 , p . 1 3 6 .

8 . B e l l a m y , L. J .
The I n f r a - r e d S p e c t r a o f C o m p l e x M o l e c u l e s ,
M e t h u e n & Co. L t d . , L o n d o n , 19 5 If", P^ 2 2 7 •

9.

B e r s c h , H. W. , a n d Do p p, W.
P r i i f u n g e i n i g e r A l k a l o i d e _in_
v i t r o a u f T u b e r k u l o s t a s e , P h a r m a z i e , 5, H e f t 2, 7 7 - 7^
(1955) •

10. B e r t h o , A.
K u r c h i - A l k a l o i d e , V I I M i t t e i l u n g : Di e K o n s t i t u t i o n d e s C o n e s s i n s und d e s C o n k u r c h i n s , B e r . , 5 6 9 , 1 - 1 6
(1950) .
11.

Bertho, A.,
cholestene,

a n d B a u m g a r t , H.
Uber 3 - p - D i m e t h y l a m i n o B e r . , 581 , II4-O-U4-8 ( 1 9 5 3 ) .

12.

Th e H o l y B i b l e , K i n g J a m e s V e r s i o n , The N a t i o n a l
P r e s s , P h i l a d e l p h i a , II K i n g s , 20, 71.

13.

B i s h o p , C. J . , a n d M a c D o n a l d , R . E .
P la n ts for A n tib ac te rial Substances,
260-2 69 (1951)120

Bible

A Survey of H igher
Can. J . B o t a n y , 2 9 ,

121
ll+. B o r s c h e , W. , a n d Pe i t z s c h , W.
U n t e r s u c h u n g e n lib e r d i e
B e s t a n d t e i l e d e r Kaw aw ur z e 1. X , r Ube r Kawa i n und D i h y d r o —
k a w a i n , B e r . , 6 3 , 21+11+-21+1 7 ( 1 9 3 0 ) .
15 .

B r e y e r - B r a n d w i j k , M. G.
La c h i m i e d e s f e u i l i e s d u S o l a n u m
P s e u d o c a p s i c u m , B u l l . S c i . P h a r m . 3 6 , 51+1-550 ( 1 9 2 9 J 1

16 .

B r i g g s , L. H . , H a r v e y , W. E . , L o c k e r , R . H . ,
W. A . , a n d S e e l y e , R . N.
Solanum A l k a l o i d s .
S o l a s o d i n e , J . Chem. S o c . , 1 9 5 0 , 3 0 1 0 - 3 0 2 0 .

17.

B r o d e r s e n , R . , a n d K j a e r , A.
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id.

C a m p b e l l , H. A . , a n d L i n k , K. P .
S t u d i e s on t h e Hemor­
r h a g i c S w e e t C l o v e r D i s e a s e IV.
Th e I s o l a t i o n a n d C r y s t a l ­
l i z a t i o n of t h e H e m o r r h a g i c A g e n t , J . B i o l . Chem., 138,
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P a r t V.

The A n t i b a c t e r i a l A c t i v i t y
Acta pharmacol. t o x ic o l.,

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80.

I b i d . , p.

li+9.

8i .

I b i d . , p.

1 71+ -

82.

S c h n e l l , L. O . , a nd T h a y e r , J . D. i n K a r e l , L . , and R o a c h ,
E. S . , E d i t o r s .
A D i e t i_on a r y o Q An t i b I o s. i s_, Col umb i a
U n i v e r s i t y P r e s s , New Y o r k , 1 9 5 1 , p . 2 7 2 .

83.

S c h o e n h e i m e r , R . , a nd E v a n s ,
Ep i a l 1 o c h o 1 e s t e r o 1 , J . B i o l .

E. A.
A 1 1o c h o 1e s t e r o 1 a nd
C h e m . , 111+, 5 6 7 - 5 8 2 ( 1 9 3 6 ) .

81+. S c h r e i b e r , K.
S o l a n u m A l k a l o i d e IV M i t t . : lib e r d i e
G l y k o a l k a l o i d e e i n i g e r W i 1d k a r t o f f e 1n d e r s e r . Commer s o n i a n a , D i e P h a r m a z i e , J_0, H e f t 6, 3 7 9 - 3 8 6 ( 1 9 5 5 ) .
85.

T h i e s , H . , a nd R e u t h e r , F. W.
R e a g e n t f o r A l k a l o i d s on
P a p e r C h r o m a t o g r a m s , N a t u r w i 3 s e n s c h a f t e n , 1+1 , 2 3 0 (1951+);
C . A . 1+9, 1+938 h ( 1 9 5 5 ) -

86.

V e l d s t r a , H . , and H a v i n g a , E.
On t h e P h y s i o l o g i c a l A c t i v ­
i t y o f U n s a t u r a t e d L a c t o n e s , E n z y m o l o g i a , _1_1_, 3 7 3 - 3 8 0
( 191+3-191+5) •

8 7 . W a t t , J . M. , H e i m a n n , H. L . , and M e l t z e r , E.
o f S o l a n u m p s e u d o c a p s i c u m on t h e C i r c u l a t o r y
Med. A s s . " S . A f r i c a , _U_, 298 ( 1 9 2 8 ) .

The A c t i o n
System, J.

8 8 . W a t t , J . M. , He I ma n n , H. L . , a n d M e l t z e r , E.
The A c t i o n
o f an A l k a l o i d a l P r o d u c t f rom t he L e a f of Solanum p s e u d o c a p s i c u m L . , J . P h a r m a c o l . 39_, 3 8 7 - 3 9 5 ( 1930 ) .

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W a t t , J . M. , H e i m a n n , H. L . , a n d E p s t e i n , E .
Solanocaps i n e - - A n A l k a l o i d w i t h a C a r d i a c A c t i o n , Q u a r t . J . Pharm.
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61+9-656 ( 1 9 3 2 ) .

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1929, p . 1092.

9 1 . W i l l i a m s , T. I .
L o n d o n , 191+7, p .
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D r u g s f r o m P l a n t s , S i g ma B o o k s ,
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W i n t e r , A. G . , a n d W i l l e k e , L.
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190-191 ( 1 9 5 2 ) .

9 3 . W o l f , M. J . , a n d D u g g a r , B, M.
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APPENDICES

APPENDIX I
ANTIBACTER IAL ACTIVITY
B i o a s s ay
The
for

s a m p l e s f r o m S o 1 anum p s e u d o c a p s i cum L_. w e r e

activity

a g ain st Mycobacterium t u b e r c u l o s i s ,

a n d D i p 1o c o c c u s p n e u m o n i a e , T y p e

III,

N.I.H.,

tested

s t r a i n H-37,

using

the

serial

d i l u t i o n method:
All

cold-water

t a m i n a t e d were
filter.

and a n y e t h a n o l

sterilized

Hot-water

by making
agar,

serial

pH 7 . 0 .

position,
with

in l i q u e f i e d

and a f t e r

solidification

o f M.

the

long

37°C.

the

the highest
active

the

cultures.

After

dilution
extracts

extract

t u b e s we r e

inoculated

tuberculos i s ,

To p r e v e n t

strain

evaporation
t u b e s were

s i x weeks of

incubation

If t h e y were

inhibited,

i n w h i c h no g r o w t h o c c u r r e d w a s r e c o r d e d .
were

10% w h o l e

extracts

beef

in a s l a n t i n g

screw-capped

c u l t u r e s were examined.

to which

alcoholic

incubation period

a Seitz

properties

glycerol

to harden
the

t o be c o n ­

by a u t o c l a v ing.

antibacterial

The a g a r wa s a l l o w e d

used fo r making

agar

dilutions

sterilized

for

found

them t h r o u g h

g r o wn o n P e t r a g n a n i 1 s me d i u m .

during

Th e

tested

an 1 8 - to -l+ 0 -d a y -o Id c u l t u r e

H-37,

at

by p a s s i n g

e x t r a c t s were

The e x t r a c t s w e r e

extracts

ten retested

on g l y c e r o l

beef ex tra ct

sh eep b l o o d had b e e n a dded.

a l l o w a n c e had
129

t o be made f o r

the

With the
inhibitory

130
effect

of

the

alcohol.

s o l v e n t was f o u n d

tubercle

bacillus

extracts

i n w h i c h t h e r e w a s no g r o w t h a t

than
of

t h i s were

the

a dilution

considered

extracts

N.I.H.,

at

This

against

active.

was d e t e r m i n e d by maki ng
(F.D.A.

of

1 8 - t o - 2 l+ -h o u r b r o t h

formula).

after

incubation for

I t was f o u n d
at

a 1:8 d i l u t i o n .

inhibition

greater

ant i b a c t e r i a l .

that

1:1+0 and t h e r e f o r e

serial

culture
the

of

dilutions

activity
III,

in n u t r i e n t

the organism per

dilutions.

1. 0 ml.
100 m l .

R e s u l t s we r e
3 7 ° C.

i n h i b i t e d D i p l o c o c c u s pn eum on i a e

C o n s e q u e n t 1y , o n l y e x t r a c t s
than th is

only

inoculated with

2l+ and 1+8 h o u r s a t

ethanol

the

a d ilu tio n higher

The a n t i b a c t e r i a l

The b r o t h wa s

o f medium j u s t b e f o r e maki ng
read

inhibit

D i p l o c o c c u s p n e u m o n i a e , Type

broth
an

of

to

s h o w i n g an

d i l u t i o n were r e c o r d e d

as b e i n g

APPENDIX I I
EXPRESSION AND CALCULATION OF ANTIBACTERIAL ACTIVITY
Th e

specific

expressed
liliter

as the

bioassay

dilution

of

1 /2 5 6 o f

the

g./m l.).

smallest

of c u lt u r e

Examp 1e :
for

antibacterial

a c t i v i t y of e a c h

A 1 per

cent

and was f o u n d
1:256.

inhibited bacterial

(w/v)
to

sample

inhibit

sample

A formula for

(which

bacterial
this

in t h i s

calculating

the

growth.

s o l u t i o n wa s s e n t

The c o n c e n t r a t i o n a t

original

( ft ) p er m i l ­

num ber o f m i c r o g r a m s

b r o t h which

s a mp l e wa s

growth at

solution

a

is

c a s e wa s 0 . 0 1

activity

is given

b e 1ow:
Spec i f ic a n t i b a c t e r i a l
C

3

D
For

= concentration

of

activity

original

= maximum o b s e r v e d d i l u t i o n
the

above example

sample

in p e r

inhibiting

cent

~

s

x 10^
D

1 x 1 0^
256

131

(w/v).

bacterial

then:
C

vml .

(in

39

growth.