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Phenelzine Sulfate
PH ENELZINE SU LF ATE
Reviewed b y L. Chafetz
383
ROBERT E. O A L Y
CON TEN T S
Analytical Profile
1. 2.
3.
4.
5. 6.
7,
-
Phenelzine S u l f a t e
Description 1.1 N a m e , F o r m u l a , M o l e c u l a r W e i g h t 1 . 2 A p p e a r a n c e , C o l o r , Odor Ph y s i ca 1 P r o p e r t i es 2 . 1 I n f r a r e d Spectrum 2.2 N u c l e a r Maqnetic Resonance Sp ect ru m 2 . 3 Mass S p e c t r u m 2.4 Ultraviolet Spectrum 2.5 D i f f e r e n t i a l Thermal A n a l y s i s 2.6 Thermogravimetric Analysis 2.7 M e l t i n g Range 2.8 S o l u b i l i t y 2.9 X-Ray D i f f r a c t i o n Powder A n a l y s i s Synthesis Stability Metabolism Methods o f A n a l y s i s 6 . 1 Spectrophotometric Analysis 6.2 Polarography 6.3 T i t r i m e t r y 6.4 Chromatography 6 . 4 1 Gas C h r o m a t o g r a p h y 6.42 Thin Layer Chromatography 6.5 S p o t T e s t s References
384
PHENELZINE SULFATE
1.
DescriDtion
N a m e , Formula, M o l e c u l a r Weight Phenelzine S u l f a t e i s B-phenylethylhydrazine dihydrogen s u l f a t e . I t i s a l s o known as p h e n e t h y l h y d r a z i n e s u l f a t e . 1.1
A p p e a r a n c e , C o l o r , Odor White t o y e l l o w i s h w h i t e c r y s t a l l i n e powder h a v i n g a c h a r a c t e r i s t i c o d o r .
1.2
2.
Physical Properties 2.1
I n f r a r e d Spectrum The i n f r a r e d s p e c t r u m ( F i g u r e 1) of p h e n e l z i n e s u l f a t e was d e t e r m i n e d a s a K B r p e l l e t i n a P e r k i n - E l m e r model 6 2 1 s p e c t r o p h o t o m e t e r . The s p e c t r u m o b t a i n e d i s i d e n t i c a l t o t h a t o f p h e n e l z i n e s u l f a t e w h i c h a p p e a r s as S a d t l e r i n f r a r e d s p e c t r u m #24825. T h e broad band a t a b o u t 2500 c m . - ’ may b e a t t r i b u t e d t o t h e hydrazine s a l t . The b a n d s a t 758 c m . - ’ a n d 705 c m . - ’ may b e a t t r i b u t e d t o t h e m o n o s u b s t i t u t e d benzene moiety. N u c l e a r Magnetic Resonance Spectrum The NMR s p e c t r i i m o f p h e n e l z i n e s u l f a t e w a s d e t e r m i n e d i n a V a r i a n A-60 i n s t r u m e n t e m p l o y i n g DMSO a s t h e s c l v e n t ( F i g u r e 2 ) . The spectrum d i s p l a y e d s i n g l e t s a t 6 7.75 (hydrogen bonded t o n i t r o g e n ) and a t 6 7.2 (aromatic). O t h e r a b s o r p t i o n w a s c e n t e r e d around 6 3.0 ( m e t h y l e n e bonded t o m e t h y l e n e and N H ) and 6 2 . 5 (methylene bonded t o methylene and p h e n y l ) D e u t e r i u m e x c h a n g e ( F i g u r e 3 ) w a s p e r f o r m e d on t h e s a m p l e . The a b s o r p t i o n band a t 6 7 . 7 5 d i s a p p e a r e d v e r i f y i n g t h e a s s i g n m e n t o f t h i s band t o hydrogen bonded t o n i t r o g e n . 2.2
.
2.0 100
0 5000
2.5
3.0
4.0 5 . 0
5.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
I
I
I
4000
3000
2000
1800
1700
1600
W A V E NUMBERS I N C Y
Figure 1
-
-'
1500
1400
1300
1200
I
-
11.0 12.0 13.0 14.0 15.0 I
-
I
.
-
1100
1000
.
.
.
.
I
I
1
1
1
1900
10.0 1
900
I n f r a r e d Spectrum o f P h e n e l z i n e S u l f a t e .
800
700 640
500
400
300
z 00
200
0 cpr
rn
C
r
n b
--I
rn
I 8.0
I
I
7.0
I
1 6.0
I
1 5.0
I
1
4.0
I
I
3.0
I
I 2.0
I
I
1.0
F i g u r e 2 - NMR S p e c t r a of P h e n e l z i n e S u l f a t e
I
0 PPm
x
500
1
1
8.0
I
300
400
1
7.0
I
I
6.0
I
I 5.0
100
200
I
I
4.0
I
I
3.0
1
I 2.0
I
1 1.0
I
0 PPm
F i g u r e 3 - NMR S p e c t r a of P h e n e l z i n e S u l f a t e - D e u t e r i u m Exchange
PHENELZINE S UL F A T E
2.3
Mass S p e c t r u m A m a s s s p e c t r u m of p h e n e l z i n e s u l f a t e could not be obtained d e s p i t e s e v e r a l attempts. An A E I 902 w a s employed w i t h a n i o n i z i n g v o l t a g e of 7 0 ev and a t a t e m p e r a t u r e of 1 0 0 ° C . On r u n ning t h e material d i r e c t l y i n t h e probe, a spect r u m w h i c h showed l a r g e a m o u n t s of s u l f u r d i o x i d e resulted. Attempts a t running t h e f r e e base proved f r u i t l e s s a l s o . The sample w a s i n t r o d u c e d i n t o t h e mass s p e c t r o m e t e r t h r o u g h t h e h e a t e d i n l e t b u t t h e r m a l d e c o m p o s i t i o n o c c u r r e d and no spectrum w a s obtained. 2.4
U l t r a v i o l e t Spectrum R a j e s w a r a n a n d K i r k (I) r e p o r t e d h max ( E 1 8 6 2 ) of 258 nm. a n d X min o f 2 2 8 nm. i n 50% ethanol. Phenelzine s u l f a t e (0.0541% i n 50% e t h a n o l ) when s c a n n e d i n a C a r y 1 4 s p e c t r o p h o t o m e t e r b e t w e e n 350 and 2 2 0 nm. ( F i g u r e 4 ) , e x h i b i t e d t h r e e p e a k s a t 2 5 2 , 258 a n d 263 nm. w i t h A max a t 258 nm. ( a = 0 . 7 9 8 : F 1 8 6 9 ) . 2.5
D i f f e r e n t i a l Thermal Analysis
A d i f f e r e n t i a l thermoqram h a s been
o b t a i n e d i n a Dupont m o d e l 9 0 0 DTA I n s t r u m e n t e m p l o y i n g a DSC c e l l . Two e n d o t h e r m s , o n e a t 134°C. ( c r y s t a l l i n e c h a n g e ) a n d t h e s e c o n d a t 1 7 2 ° C . ( m e l t i n g ) were o b s e r v e d ( F i g u r e 5 ) . H e a t i n g r a t e was 10' C . / m i n u t e . The d e t e r m i n a t i o n w a s c a r r i e d out i n a n i t r o g e n a t m o s p h e r e . A l t e r a t i o n i n t h e r a t e of h e a t i n g w o u l d t e n d t o s h i f t t h e endotherms. Thermogravimetric Analysis A thermogravimetric analysis was perf o r m e d ( F i g u r e 6 ) i n a D u p o n t m o d e l 9 5 0 TGA Instrument. The m e a s u r e m e n t was p e r f o r m e d u n d e r n i t r o g e n sweep a t a h e a t i n g r a t e o f 1 0 " C . / minute. N o w e i g h t loss w a s n o t e d u n t i l a b o u t 165" C . ( i n i t i a t i o n o f m e l t i n g ) . A f t e r t h i s p o i n t , w e i g h t w a s r a p i d l y lost. T h i s i n f o r m a t i o n t o g e t h e r w i t h t h e DTA i n d i c a t e s t h a t t h e 2.6
389
R O B E R T E. DALY
0. a
0. :
0.6
0.5
rn
*
c, Y
0.4
eq
e
0
2
0.3
pl
0.2
0.1
210
Figure 4
-
WAVELENGTH
320
W Spectrum o f Phenelzine S u l f a t e
390
0x3-
0aN3
PHENELZINE SULFATE
JV-
391
0 0
rn 0
2 3
0 '0
0
2 0
N
crj k
a r
a:
k -!
5
a:
q"
2 Y 0,
0
0 05
0 \o
0
w
0
rn
0
0
5 k
rn 0
E e,
k
c E m
8-4
ROBERT E. D A L Y
392
a 0 v,
0 v,
w
0
*0
m
0 v)
0 0
m 0 v)
w
0
0
w
2
0
1
0 0
a v,
0
a,
c, m w
a, C
N
.d
C
a,
rl
II)
a,
c Y 4
C
h m
q" rl
a:
5 k
P
am:
s 2
PHENELZINE SULFATE
e n d o t h e r m a t 134O C . w a s d u e t o a n o n d e s t r u c t i v e a l t e r a t i o n i n t h e molecule (crystal change). 2.7
M e l t i n g Range The m e l t i n g r a n g e h a s b e e n r e p o r t e d ( 2 ) t o be 1 6 4 t o 1 6 8 " C . 2.8
Solubility Phenelzine s u l f a t e i s soluble i n water and p r a c t i c a l l y i n s o l u b l e i n e t h a n o l , c h l o r o f o r m and e t h e r . 2.9
X-Ray D i f f r a c t i o n Powder A n a l y s i s R a j e s w a r a n a n d K i r k (1) h a v e r e p o r t e d x-ray d i f f r a c t i o n data €or p h e n e l z i n e s u l f a t e . The powdered s a m p l e which h a d b een a p p r o p r i a t e l y m o u n t e d on a r o t a r y s p e c i m e n h o l d e r , w a s s u b j e c t e d t o copper k-alpha r a d i a t i o n i n a Norelco G e i g e r C o u n t e r X-Ray S p e c t r o m e t e r . The d i f f r a c t i o n p a t t e r n s w e r e o b t a i n e d over a n a r c of 4 5 " . The i n s t r u m e n t w a s c a l i b r a t e d o v e r t h i s r a n g e , u s i n g powdered q u a r t z c r y s t a l s , a n d t h e " d " v a l u e s computed. Values are g i v e n i n T a b l e I . Table I
-
Ild" D i s t a n c e s U s i n g
C o p p e r K-Alpha Distances
3.
-
18.9, 3.24,
9.85, 2.79
&
Radiation
6.51, 5.42, 2.44
4.89,
3.90,
Synthesis
B i e l ( 3 ) h a s s y n t h e s i z e d p h e n e l z i n e by r e a c t i n g h y d r a z i n e h y d r a t e and p h e n e t h y l bromide under r e f l u x €or 7 h o u r s . A y i e l d of 77% w a s o b t a i n e d . Sacha and M a l a s n i c k i ( 4 ) condensed p h e n e t h y l c h l o r i d e w i t h hydrazine under r e f l u x i n anhydrous e t h a n o l i n a n i t r o g e n atmosphere €or 1 2 hours. T h e s o l u t i o n of t h e f r e e b a s e w h i c h w a s o b t a i n e d a f t e r removal o f h y d r a z i n e H C 1 b y f i l t r a t i o n was t r e a t e d w i t h s u l f u r i c a c i d i n anhydrous e t h a n o l 393
ROBERT E. DALY
.
(l.l/l W/W) A y i e l d of 2 1 % o f P - p h e n y l e t h y l hydrazine dihydrogen s u l f a t e w a s obtained. B i e l , e t . aZ. ( 5 ) h a s r e p o r t e d t h a t a r a l k y l a t i o n of hydrazine with primary a r a l k y l h a l i d e s a f f o r d s mono- ( a r a l k y l ) - h y d r a z i n e s i n y i e l d s of 6 0 t o 75% p r o v i d e d a t h r e e t o f i v e f o l d molar excess o f h y d r a z i n e h y d r a t e i s employed. Production of d i p h e n y l e t h y l h y d r a z i n e has been r e p o r t e d ( 6 ) as a by-product i n t h e r e a c t i o n .
0-
Reflux 7 hours
>\:2;::;:::;:
anhyd.
EtOH
1
O H P - C H Z - N H - N H ~
12R'/;d.
NP atm.
CH2-CH2C1 + NH~-NHP*H~O
4.
H2S04 : E t O H
(l*l:l)
Sulfate
Stability
P h e n e l z i n e base i s e a s i l y o x i d i z e d . Decompos i t i o n o c c u r s when b a s i f i e d s o l u t i o n s a r e e x t r a c t e d , probably due t o autooxidation of t h e base. S c h l i t t , e t . aZ. ( 7 ) h a v e r e p o r t e d t h e a u t o o x i d a t i o n of p h e n e l z i n e s u l f a t e i n pH 5 . 9 p h o s p h a t e buffer. A u t o o x i d a t i o n i n pH 6 . 5 , 0 . 1 M s o d i u m c h l o r a t e a t 37' C. h a s a l s o b e e n r e p o r t e d ( 8 ) . D e c o m p o s i t i o n p r o d u c t s were n o t c h a r a c t e r i z e d . 5.
Metabolism D r a b n e r a n d Schwerd ( 9 ) i d e n t i f i e d u n c h a n g e d 394
PHENELZINE SULFATE
p h e n e l z i n e and p h e n y l a c e t y l g l u t a m i n e i n t h e u r i n e o f humans who h a d r e c e i v e d p h e n e l z i n e s u l f a t e . C l i n e s c h m i d t , e t . a l . ( 1 0 , 11) s t u d i e d t h e in v i t r o b i o t r a n s f o r m a t i o n a s w e l l as t h e in v i v o metabolism of C"-phenelzine i n rats. Phenylaceti c a c i d w a s i d e n t i f i e d as t h e m a j o r m e t a b o l i t e . F i s c h e r , e t . al. ( 1 2 ) h a v e d e s c r i b e d t h e e f f e c t o f p h e n e l z i n e on t h e e x c r e t i o n of 6phenethylamine. 6.
Methods of A n a l y s i s 6.1
Spectrophotometric Analysis
6 . 1 1 F e i g l (13) has r e p o r t e d t h e reduct i o n of l i t h i u m and s o d i u m molybdophosphotungs t a t e s o l u t i o n by h y d r a z i n e d e r i v a t i v e s . T h i s r e a c t i o n h a s b e e n a d a p t e d t o t h e d e t e r m i n a t i o n of p h e n e l z i n e s u l f a t e r a w m a t e r i a l o r t a b l e t s by Bose and V i j a y v a r g i y a (14). R e d u c t i o n i s c a r r i e d o u t i n a l k a l i n e s o l u t i o n with t h e production of a d e e p b l u e c o l o r w h i c h is m e a s u r e d a t 6 5 0 nm.
6.12 A c o l o r i m e t r i c method b a s e d on t h e formation of t h e hydrazone w i t h v a n i l l i n has been d e v i s e d ( 1 5 ) . The r e a c t i o n i s c a r r i e d o u t i n 0.5 N m e t h a n o l i c h y d r o c h l o r i c a c i d . Absorbance o f t h e y e l l o w s o l u t i o n i s r e a d a t 4 0 0 nm. S i n c e p h e n e l z i n e d e g r a d e s by o x i d a t i v e d e s t r u c t i o n of t h e h y d r a z i n e f u n c t i o n , t h i s method i s s t a b i l i t y indicating
.
Polarography The h a l f wave p o t e n t i a l versus s t a n d a r d calomel e l e c t r o d e ) of t h e a c e t o n e d e r i v a t i v e of p h e n e l z i n e w a s r e p o r t e d ( 7 ) as - 1 . 3 3 V i n pH 5 . 9 p h o s p h a t e b u f f e r ( 0 . 1 M KH~POI, : N a 2 H P O 4 * 2 H 2 O ) employing a 0 . 0 1 % g e l a t i n s o l u t i o n as a maximum s u p p r e s s o r . The d i f f u s i o n c u r r e n t Four elecc o n s t a n t ( I d ) w a s approximately 2.5. t r o n s w e r e t r a n s f e r r e d f o r t h e r e d u c t i o n of t h e h y d r a z o n e d e r i v a t i v e . W a v e h e i g h t was p r o p o r t i o n a l t o c o n c e n t r a t i o n i n t h e r a n g e of 2 . 5 x lo-' t o 2.5 x M i n pH 5 . 9 p h o s p h a t e b u f f e r . 6.2
3Y.5
ROBERT E . D A L Y
6.3
Titrimetry P r o c e d u r e s b a s e d on t h e r e a c t i o n o f phenelzine s u l f a t e with iodine i n bicarbonate s o l u t i o n ( 2 ) and i n sodium h y d r o x i d e ( 1 6 ) and w i t h b r o m a t e and bromide i n a c i d s o l u t i o n ( 1 6 ) w i t h s u b s e q u e n t t i t r a t i o n of e x c e s s r e a g e n t ( i o d i n e o r bromide c o n v e r t e d t o i o d i n e ) i n a c i d s o l u t i o n w i t h t h i o s u l f a t e have been r e p o r t e d . B a s i c a l l y , t h e r e a c t i o n i n v o l v e d may be i l l u s t r a t e d a s follows:
Radecka and Nigam ( 1 7 ) have r e p o r t e d t h e d i r e c t t i t r a t i o n o f p h e n e l z i n e w i t h N-bromosuccinimide i n a c i d i c s o l u t i o n employing m e t h y l r e d a s t h e indicator. However, t h e i r r e s u l t s show a 5% p o s i t i v e b i a s . This b i a s is a t t r i b u t e d t o a l l y l i c b r o m i n a t i on by N -b r omosu c c i n i m i d e
.
6.4
Chromatography
Gas Chromatography C a r d i n i , e t . a l . (18, 1 9 ) and McMartin and S t r e e t ( 2 0 ) h a v e r e p o r t e d g a s chroma t o g r a p h i c methods which a r e r e p u t e d t o be quantitative. However, no q u a n t i t a t i v e d a t a a r e g i v e n . C o n d i t i o n s a r e r e p o r t e d i n T a b l e 11. 6.41
T h i n Layer Chromatography Q u a l i t a t i v e methods f o r t h e i d e n t i f i c a t i o n of p s y c h o t r o p i c a g e n t s have b e e n r e p o r t e d ( 2 1 , 2 2 , 2 3 , 2 4 , 2 5 ) . R f v a l u e s a r e summarized i n T a b l e 111. 6.42
6.5
Spot T e s t s A v a r i e t y of s p o t t e s t s have been re-
ported ( 2 6 ) .
T h e s e are summarized i n T a b l e 396
v.
T A B L E I1 - P a r a m e t e r s f o r G a s C h r o m a t o g r a p h y Ref. 18
19 w
c 4
20
Column g l a s s column 1 . 8 M x 2 mm.; 2 % GE-SE 3 0 on A e r o p a k 3 0 , 80100 m e s h
Column Temp.
Carrier Gas
program 70-250' a t 10.4O/
N2
Flow Rate
50 m l . / min.
Detector
FID
Detector Te m p
.
220'
C.
Injector Temp. 250'
C.
minute
Q
Stainless steel 140' c o l u m n 1 . 2 M. x 2 mm.; 15%c a r b o w a x 20 M o n C h r o m o s o r b W alkalinized w i t h 5% KOH
C.
Stainless steel 140' c o l u m n 6' x 1/8" ; 2 % SE 3 0 + 0 . 1 % t r i s t e a r i n on s i l a n i z e d acid w a s h e d C h r o m o sorb W
C.
He
40 m l . / min.
FID
250'
C.
I rn
z
rn
P z
rn
m C
r
n
>
2 N2
3 0 ml./ min.
FID
ROBERT
TABLE I11
-
E. D A L Y
S u p p o r t , S o l v e n t System and Rf
V a l u e s f o r TLC of P h e n e l z i n e S u l f a t e Reference
Support
Solvent System
Rf
(X
21
silica gel G
chloroform: m e t hano 1 (8:2)
21
silica gel G
chloroform: acetone: d i e t h y lamine ( 5 : 4: 1)
88
21
silica gel G
cyclohexane: chloroform: d ie t h y l a m ine ( 4 :5 :1)
62
22
silica gel G impregnated with 0 . 1 M N aOH
hexane:benzene: diethylamine ( 7 5 : 1 5 : 10)
51
22
silica gel G impregnated with 0.1 M N aOH
methanol
72
22
silica gel G imp re gn a t ed with 0 . 1 M NaOH
acetone
75
22
silica gel G impregnated with 0 . 1 M NaHSO
methanol
41
398
100
100)
PHENELZINE SULFATE
TABLE I11 f c o n t . 1
R e f er e n ce
Support
Solvent System
Rf
( x 100)
silica gel G impregnated with 0 . 1 M NaHSO
95% ethanol
25
silica gel G activated a t 110" C. f o r 30 m i n u t e s
chloroform: m ethano1
37
23
silica gel G a c t i v a t e d as above
chloroform: methanol : 2 0 % ammonium hydroxide ( 2 : 1:l)
56
24
silica gel G activated at 100' C . f o r 1 hour
methanol:12 N ammon i um hydroxide (100: 1.5)
74
24
silica gel G impregnated with 0 . 1 N NaOH and activated a s above
cyclohexane: diethylamine: benzene (75: 20: 15)
44
24
s i l i c a g e l G acetone impregnated and a c t i v a t e d as above
65
24
s i l i c a g e l G chloroform: impregnated methanol and a c t i v a t e d ( 9 0 : 10) as above
75
22
+ ,
23
(1:l)
399
ROBERT E. DALY
TABLE I11 ( c o n t . )
Reference
Support
Solvent System
Rf
(X
100)
24
silica gel G
benzene: ethanol: 1 2 N ammonium hydroxide ( 9 5 : 15: 5 )
50
25
silica gel G imp re g n a t e d with 0 . 1 N KHSO 4
95% e t h a n o l
49
25
silica gel G impregnated w i t h 0.1N
cyclohexane: benzene : d i et h y lamine ( 7 5 : 15: 10)
52
silica gel G imp r e g n a t e d with 0 . 1 N
methanol
72
silica gel G impregnated with 0.1 N
acetone
75
25
silica gel G imp r e g n a t e d with 0 . 1 N KHSO I,
methanol
62
25
silica gel G impregnated with 0 . 1 N NaOH
methyl acetate
53
N aOH
25
N aOH
25
N aOH
400
PHENE LZI NE SULFATE
TABLE I11 ( c o n t . ) Reference 25
Support
Solvent System
chromedi a n-butanol: (Whatman #41) 5% c i t r i c impregnated acid w i t h 5% s o d i u m d i h y d r og e n citrate (some K2HP04 a d d e d t o prevent tailing)
Rf (x 100) 51
ROBERT E. DALY
TABLE I V
-
Reagents and Reactions
F o r TLC P r o c e d u r e s Reagent
Reaction
21
Folin-Ciocalteau's Reagent
blue violet with heating
21
5% phosphomolybdic a c i d i n e t h a n o l f o l l o w e d by exposure of t h e p l a t e t o ammonia v a p o r
blue
22
1%i o d i n e i n m e t h a n o l
brown
23
50 m l . o f 0 . 2 % ninhydrin i n methanol + 1 0 m l . of g l a c i a l a c e t i c a c i d + 2 m l . of
r o s e orange
23
1 0 % F e r r i c c h l o r i d e ; 1%
fluorescent blue
23
S a t u r a t e d ammonium molybdate; s a t u r a t e d oxalic acid
celeste
24
Fo l i n - C i o c a l t e a u ' s Reagent
b l u i s h a t RT, b l u e when heated t o
Reference
2,4,6-trimethylpiperidine f l u o r e s c e i n : ammonia
100'
c.
24
Mandelin s R e a g e n t
pink a t RT, f l e s h a t 100' C. , f l u o r e s c e n t a f t e r heating
24
C i nn ama l d e h y d e R e a g e n t 5 m l . cinnamaldehyde + 95 m l . ethanol + 5 m l . conc. HC1
y e l l o w a t RT
402
PHENELZINE SULFATE
TABLE I V ( c o n t . )
Reference
Reagent
Reaction
24
0.25 g . p-dimethylaminobenzaldehyde + 5 g . of 85% p h o s p h o r i c a c i d + 20 m l . of water
lemon chromo after heating t o 1 0 0 ' C. f o r 1 0 min. and t h e n exposed t o ammonia v a p o r
24
Furf u r a l r e a g e n t s o l ' n A.) 1 m l . f u r f u r a l i n 99 m l . of acetone s o l ' n B . ) 4 m l . of c o n c . H 2 S 0 4 i n 96 m l . o f a c e t o n e
brown a t 1 0 0 '
25
P l a t e examined u n d e r s h o r t w a v e W l i g h t and t h e n s p r a y e d w i t h t h e following sequence of r e a g e n t s :
C.
iodine
brown
Dragendorff ' s
orange
sodium n i t r i t e
d e e p brown
i odop l a t in a t e until a color develops
variously colored
403
ROBERT E. DALY
TABLE V
-
Spot Tests f o r Phenelzine S u l f a t e
Reagent
Response 0 . 5 mg.
2 0 1-14.
Froehde ' s Reagent
blue* It. b l u e
blue
Mandelin ' s R e a g e n t
green blue* brown
green faint orange
Marquis ' Reagent Mecke's R e a g e n t
r e d orange* grey
pink
R e i c k a r d ' s Reagent
It. blue
It. b l u e
F l u e c k i g e r ' s Reagent
brown
V i t a l i ' s Reagent
a. 1 b.)
Was i c k y ' s R e agen t
red o r a n g e
*
=
a f t e r warming
404
- -blue orange
---
--- --
PH E N E LZ IN E SULFATE
TABLE V I
- Composition o f Spot T e s t Reagents
Reagent
Composition
Froehde I s Reagent
0 . 5 % a q u e o u s ammonium m o 1y b d a t e
Mande l i n I s R e a g e n t
0 . 5 % a q u e o u s ammonium vanadate
Marquis
1 p a r t 37% formaldehyde i n 20 p a r t s concentrated
Reagent
sulfuric acid, freshly prepared before use Mecke ' s R e a g e n t
0 . 5 % aqueous s e l e n i o u s a c i d
Reickard I s Reagent
1%a q u e o u s sodium t u n g s t a t e
F l u e c k i g e r ' s Reagent
0 . 5 % s o l u t i o n of t i t a n i u m d i o x i d e ( T i 0 2 1 i n concentrated sulfuric acid
V i t a l i s Reagent
a.) b.)
Wasicky s R e a g e n t
(H2SeO 3 1
fuming n i t r i c a c i d
3% potassium hydroxide
i n ethanol
1 0 % s o l u t i o n o f p-dimethylaminobenzaldehyde i n g l a c i a l acetic acid
305
ROBERT E. DALY
7.
References R a j e s w a r a n a n d P . L. K i r k , B u l l . N a r c o t i c s , U . N . , D e p t . S o : L 4 i l ~ f f ~ i 1r 4 s, No. I, 1 9 ( 1 9 6 2 ) .
1.
P.
2. 3. 4.
U.S.P. X V I I I . J . H . B i e l , US 3 , 0 0 0 , 9 0 3 , S e p t . 1 9 , 1 9 6 1 . A. S a c h a a n d L . M a l a s n i c k i , P o l 4 6 , 2 3 7 , O c t . 3 0 , 1 9 6 2 , C . A . 60, 4 5 4 f ( 1 9 6 3 ) . J . H . B i e l , A. E . D r u k k e r , T . H . M i t c h e l l , E . P . S p r e n g e l e r , P . A . N u h f e r , A. C . Conway and A. H o r i t a , J . A m . Chem. S o c . 8 2 , 2805 ( 1 9 5 9 ) . E . V o t o c e k a n d 0. L e m i n g e r , ColI. C z e c h . Chem. Commun. 4 , 2 7 1 ( 1 9 3 2 ) , C. A . 2 6 ,
5.
6. 7. 8. 9.
10.
11. 12.
13. 14.
15. 16. 17. 18. 19. 20.
5294 ( 1 9 3 2 ) . L . S c h l i t t , M. R i n k a n d M. von S t a c k e l b e r g , J , E Z e c t r o a n a l . Chem. 2 3 , 10 (1967). L . E . E b e r s o n a n d K . P e r s s o n , ,J. Med. Pharm. Chem. 5, 7 3 8 ( 1 9 6 2 ) . J . D r a b n e r a n d W . S c h w e r d , F r e s e n i u s ' Z. A n a l . Chem. 2 4 3 , 9 2 ( 1 9 6 8 ) . C. A . 70, 558941 ( 1 9 6 9 ) B . Van C l i n e s c h m i d t a n d A. H o r i t a , Biochem. Pharmacol. 1 8 , 1 0 1 1 (1969) Idem., i b i d . , 1021 (1969). E . F i s c h e r , B . H e l l e r , a n d A . H . Miro, A r z n e i m i t t e l - F o r s c h . 18, 1 4 8 6 ( 1 9 6 8 ) . F. F e i g l , "Spot T e s t i n O r g a n i c Analysis", E l s e v i e r Company, L o n d o n , 1 9 5 6 , p . 1 2 8 . B . C . Bose a n d R . V i j a y v a r g i y a , I n d i a n J . Pharm. 2 8 , 328 ( 1 9 6 6 ) . R. E . D a l y , u n p u b l i s h e d r e s u l t s M. M a r z a d r o a n d A. D e C a r o l i s , R e n d . I s t . S u n i t a 2 6 , 629 ( 1 9 6 3 ) . C . R a d e c k a a n d I . C. Nigam, Can. J . P h m m S c i . 2 , 17 (1966). C . C a r d i n i , V . Q u e r c i a a n d A . C a l o , :J. C h r o m a t o g . 37, 1 9 0 ( 1 9 6 8 ) . U. Q u e r c i a , C . C a r d i n i a n d A . Calo, i i ( ' l . C h i m . Farm. 1 0 7 , 3 8 3 ( 1 9 6 8 ) . C. M c M a r t i n a n d H . V . S t r e e t , J . Chromatog. 2 2 , 274 ( 1 9 6 6 ) .
.
.
406
PHENELZINE SULFATE
7.
References
21. 22. 23. 24. 25. 26.
(cont. )
E. S c h m i d , E. Hoppe, C . M e y t h a l e r a n d L.
Z i c h a , A r z n e i m i t t e Z - F o r s c h , 1 3 , 9 6 9 (1963). W . W. F i k e , A n a l . Chem. 38, 1 6 9 7 ( 1 9 6 6 ) . A . A l e s s a n d r o , F. Mari a n d S . S e t t e c a s e , F arma co , Ed. Prat. 22, 437 ( 1 9 6 7 ) . I. Z i n g a l e s , J . C h r o m a t o g . 3 1 , 405 (1967). I. S u n s h i n e , W. W . F i k e a n d H . L a n d e s m a n , J . F o r e n s i c S c i . 1 1 , 428 ( 1 9 6 6 ) . P. R a j e s w a r a n a n d P . L . K i r k , B u l l . Narcotics, U.N., Dept. S o c i a l A f f a i r s 13, No. 3, 15 ( 1 9 6 1 ) .
407
Reviewed b y L. Chafetz
383
ROBERT E. O A L Y
CON TEN T S
Analytical Profile
1. 2.
3.
4.
5. 6.
7,
-
Phenelzine S u l f a t e
Description 1.1 N a m e , F o r m u l a , M o l e c u l a r W e i g h t 1 . 2 A p p e a r a n c e , C o l o r , Odor Ph y s i ca 1 P r o p e r t i es 2 . 1 I n f r a r e d Spectrum 2.2 N u c l e a r Maqnetic Resonance Sp ect ru m 2 . 3 Mass S p e c t r u m 2.4 Ultraviolet Spectrum 2.5 D i f f e r e n t i a l Thermal A n a l y s i s 2.6 Thermogravimetric Analysis 2.7 M e l t i n g Range 2.8 S o l u b i l i t y 2.9 X-Ray D i f f r a c t i o n Powder A n a l y s i s Synthesis Stability Metabolism Methods o f A n a l y s i s 6 . 1 Spectrophotometric Analysis 6.2 Polarography 6.3 T i t r i m e t r y 6.4 Chromatography 6 . 4 1 Gas C h r o m a t o g r a p h y 6.42 Thin Layer Chromatography 6.5 S p o t T e s t s References
384
PHENELZINE SULFATE
1.
DescriDtion
N a m e , Formula, M o l e c u l a r Weight Phenelzine S u l f a t e i s B-phenylethylhydrazine dihydrogen s u l f a t e . I t i s a l s o known as p h e n e t h y l h y d r a z i n e s u l f a t e . 1.1
A p p e a r a n c e , C o l o r , Odor White t o y e l l o w i s h w h i t e c r y s t a l l i n e powder h a v i n g a c h a r a c t e r i s t i c o d o r .
1.2
2.
Physical Properties 2.1
I n f r a r e d Spectrum The i n f r a r e d s p e c t r u m ( F i g u r e 1) of p h e n e l z i n e s u l f a t e was d e t e r m i n e d a s a K B r p e l l e t i n a P e r k i n - E l m e r model 6 2 1 s p e c t r o p h o t o m e t e r . The s p e c t r u m o b t a i n e d i s i d e n t i c a l t o t h a t o f p h e n e l z i n e s u l f a t e w h i c h a p p e a r s as S a d t l e r i n f r a r e d s p e c t r u m #24825. T h e broad band a t a b o u t 2500 c m . - ’ may b e a t t r i b u t e d t o t h e hydrazine s a l t . The b a n d s a t 758 c m . - ’ a n d 705 c m . - ’ may b e a t t r i b u t e d t o t h e m o n o s u b s t i t u t e d benzene moiety. N u c l e a r Magnetic Resonance Spectrum The NMR s p e c t r i i m o f p h e n e l z i n e s u l f a t e w a s d e t e r m i n e d i n a V a r i a n A-60 i n s t r u m e n t e m p l o y i n g DMSO a s t h e s c l v e n t ( F i g u r e 2 ) . The spectrum d i s p l a y e d s i n g l e t s a t 6 7.75 (hydrogen bonded t o n i t r o g e n ) and a t 6 7.2 (aromatic). O t h e r a b s o r p t i o n w a s c e n t e r e d around 6 3.0 ( m e t h y l e n e bonded t o m e t h y l e n e and N H ) and 6 2 . 5 (methylene bonded t o methylene and p h e n y l ) D e u t e r i u m e x c h a n g e ( F i g u r e 3 ) w a s p e r f o r m e d on t h e s a m p l e . The a b s o r p t i o n band a t 6 7 . 7 5 d i s a p p e a r e d v e r i f y i n g t h e a s s i g n m e n t o f t h i s band t o hydrogen bonded t o n i t r o g e n . 2.2
.
2.0 100
0 5000
2.5
3.0
4.0 5 . 0
5.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
I
I
I
4000
3000
2000
1800
1700
1600
W A V E NUMBERS I N C Y
Figure 1
-
-'
1500
1400
1300
1200
I
-
11.0 12.0 13.0 14.0 15.0 I
-
I
.
-
1100
1000
.
.
.
.
I
I
1
1
1
1900
10.0 1
900
I n f r a r e d Spectrum o f P h e n e l z i n e S u l f a t e .
800
700 640
500
400
300
z 00
200
0 cpr
rn
C
r
n b
--I
rn
I 8.0
I
I
7.0
I
1 6.0
I
1 5.0
I
1
4.0
I
I
3.0
I
I 2.0
I
I
1.0
F i g u r e 2 - NMR S p e c t r a of P h e n e l z i n e S u l f a t e
I
0 PPm
x
500
1
1
8.0
I
300
400
1
7.0
I
I
6.0
I
I 5.0
100
200
I
I
4.0
I
I
3.0
1
I 2.0
I
1 1.0
I
0 PPm
F i g u r e 3 - NMR S p e c t r a of P h e n e l z i n e S u l f a t e - D e u t e r i u m Exchange
PHENELZINE S UL F A T E
2.3
Mass S p e c t r u m A m a s s s p e c t r u m of p h e n e l z i n e s u l f a t e could not be obtained d e s p i t e s e v e r a l attempts. An A E I 902 w a s employed w i t h a n i o n i z i n g v o l t a g e of 7 0 ev and a t a t e m p e r a t u r e of 1 0 0 ° C . On r u n ning t h e material d i r e c t l y i n t h e probe, a spect r u m w h i c h showed l a r g e a m o u n t s of s u l f u r d i o x i d e resulted. Attempts a t running t h e f r e e base proved f r u i t l e s s a l s o . The sample w a s i n t r o d u c e d i n t o t h e mass s p e c t r o m e t e r t h r o u g h t h e h e a t e d i n l e t b u t t h e r m a l d e c o m p o s i t i o n o c c u r r e d and no spectrum w a s obtained. 2.4
U l t r a v i o l e t Spectrum R a j e s w a r a n a n d K i r k (I) r e p o r t e d h max ( E 1 8 6 2 ) of 258 nm. a n d X min o f 2 2 8 nm. i n 50% ethanol. Phenelzine s u l f a t e (0.0541% i n 50% e t h a n o l ) when s c a n n e d i n a C a r y 1 4 s p e c t r o p h o t o m e t e r b e t w e e n 350 and 2 2 0 nm. ( F i g u r e 4 ) , e x h i b i t e d t h r e e p e a k s a t 2 5 2 , 258 a n d 263 nm. w i t h A max a t 258 nm. ( a = 0 . 7 9 8 : F 1 8 6 9 ) . 2.5
D i f f e r e n t i a l Thermal Analysis
A d i f f e r e n t i a l thermoqram h a s been
o b t a i n e d i n a Dupont m o d e l 9 0 0 DTA I n s t r u m e n t e m p l o y i n g a DSC c e l l . Two e n d o t h e r m s , o n e a t 134°C. ( c r y s t a l l i n e c h a n g e ) a n d t h e s e c o n d a t 1 7 2 ° C . ( m e l t i n g ) were o b s e r v e d ( F i g u r e 5 ) . H e a t i n g r a t e was 10' C . / m i n u t e . The d e t e r m i n a t i o n w a s c a r r i e d out i n a n i t r o g e n a t m o s p h e r e . A l t e r a t i o n i n t h e r a t e of h e a t i n g w o u l d t e n d t o s h i f t t h e endotherms. Thermogravimetric Analysis A thermogravimetric analysis was perf o r m e d ( F i g u r e 6 ) i n a D u p o n t m o d e l 9 5 0 TGA Instrument. The m e a s u r e m e n t was p e r f o r m e d u n d e r n i t r o g e n sweep a t a h e a t i n g r a t e o f 1 0 " C . / minute. N o w e i g h t loss w a s n o t e d u n t i l a b o u t 165" C . ( i n i t i a t i o n o f m e l t i n g ) . A f t e r t h i s p o i n t , w e i g h t w a s r a p i d l y lost. T h i s i n f o r m a t i o n t o g e t h e r w i t h t h e DTA i n d i c a t e s t h a t t h e 2.6
389
R O B E R T E. DALY
0. a
0. :
0.6
0.5
rn
*
c, Y
0.4
eq
e
0
2
0.3
pl
0.2
0.1
210
Figure 4
-
WAVELENGTH
320
W Spectrum o f Phenelzine S u l f a t e
390
0x3-
0aN3
PHENELZINE SULFATE
JV-
391
0 0
rn 0
2 3
0 '0
0
2 0
N
crj k
a r
a:
k -!
5
a:
q"
2 Y 0,
0
0 05
0 \o
0
w
0
rn
0
0
5 k
rn 0
E e,
k
c E m
8-4
ROBERT E. D A L Y
392
a 0 v,
0 v,
w
0
*0
m
0 v)
0 0
m 0 v)
w
0
0
w
2
0
1
0 0
a v,
0
a,
c, m w
a, C
N
.d
C
a,
rl
II)
a,
c Y 4
C
h m
q" rl
a:
5 k
P
am:
s 2
PHENELZINE SULFATE
e n d o t h e r m a t 134O C . w a s d u e t o a n o n d e s t r u c t i v e a l t e r a t i o n i n t h e molecule (crystal change). 2.7
M e l t i n g Range The m e l t i n g r a n g e h a s b e e n r e p o r t e d ( 2 ) t o be 1 6 4 t o 1 6 8 " C . 2.8
Solubility Phenelzine s u l f a t e i s soluble i n water and p r a c t i c a l l y i n s o l u b l e i n e t h a n o l , c h l o r o f o r m and e t h e r . 2.9
X-Ray D i f f r a c t i o n Powder A n a l y s i s R a j e s w a r a n a n d K i r k (1) h a v e r e p o r t e d x-ray d i f f r a c t i o n data €or p h e n e l z i n e s u l f a t e . The powdered s a m p l e which h a d b een a p p r o p r i a t e l y m o u n t e d on a r o t a r y s p e c i m e n h o l d e r , w a s s u b j e c t e d t o copper k-alpha r a d i a t i o n i n a Norelco G e i g e r C o u n t e r X-Ray S p e c t r o m e t e r . The d i f f r a c t i o n p a t t e r n s w e r e o b t a i n e d over a n a r c of 4 5 " . The i n s t r u m e n t w a s c a l i b r a t e d o v e r t h i s r a n g e , u s i n g powdered q u a r t z c r y s t a l s , a n d t h e " d " v a l u e s computed. Values are g i v e n i n T a b l e I . Table I
-
Ild" D i s t a n c e s U s i n g
C o p p e r K-Alpha Distances
3.
-
18.9, 3.24,
9.85, 2.79
&
Radiation
6.51, 5.42, 2.44
4.89,
3.90,
Synthesis
B i e l ( 3 ) h a s s y n t h e s i z e d p h e n e l z i n e by r e a c t i n g h y d r a z i n e h y d r a t e and p h e n e t h y l bromide under r e f l u x €or 7 h o u r s . A y i e l d of 77% w a s o b t a i n e d . Sacha and M a l a s n i c k i ( 4 ) condensed p h e n e t h y l c h l o r i d e w i t h hydrazine under r e f l u x i n anhydrous e t h a n o l i n a n i t r o g e n atmosphere €or 1 2 hours. T h e s o l u t i o n of t h e f r e e b a s e w h i c h w a s o b t a i n e d a f t e r removal o f h y d r a z i n e H C 1 b y f i l t r a t i o n was t r e a t e d w i t h s u l f u r i c a c i d i n anhydrous e t h a n o l 393
ROBERT E. DALY
.
(l.l/l W/W) A y i e l d of 2 1 % o f P - p h e n y l e t h y l hydrazine dihydrogen s u l f a t e w a s obtained. B i e l , e t . aZ. ( 5 ) h a s r e p o r t e d t h a t a r a l k y l a t i o n of hydrazine with primary a r a l k y l h a l i d e s a f f o r d s mono- ( a r a l k y l ) - h y d r a z i n e s i n y i e l d s of 6 0 t o 75% p r o v i d e d a t h r e e t o f i v e f o l d molar excess o f h y d r a z i n e h y d r a t e i s employed. Production of d i p h e n y l e t h y l h y d r a z i n e has been r e p o r t e d ( 6 ) as a by-product i n t h e r e a c t i o n .
0-
Reflux 7 hours
>\:2;::;:::;:
anhyd.
EtOH
1
O H P - C H Z - N H - N H ~
12R'/;d.
NP atm.
CH2-CH2C1 + NH~-NHP*H~O
4.
H2S04 : E t O H
(l*l:l)
Sulfate
Stability
P h e n e l z i n e base i s e a s i l y o x i d i z e d . Decompos i t i o n o c c u r s when b a s i f i e d s o l u t i o n s a r e e x t r a c t e d , probably due t o autooxidation of t h e base. S c h l i t t , e t . aZ. ( 7 ) h a v e r e p o r t e d t h e a u t o o x i d a t i o n of p h e n e l z i n e s u l f a t e i n pH 5 . 9 p h o s p h a t e buffer. A u t o o x i d a t i o n i n pH 6 . 5 , 0 . 1 M s o d i u m c h l o r a t e a t 37' C. h a s a l s o b e e n r e p o r t e d ( 8 ) . D e c o m p o s i t i o n p r o d u c t s were n o t c h a r a c t e r i z e d . 5.
Metabolism D r a b n e r a n d Schwerd ( 9 ) i d e n t i f i e d u n c h a n g e d 394
PHENELZINE SULFATE
p h e n e l z i n e and p h e n y l a c e t y l g l u t a m i n e i n t h e u r i n e o f humans who h a d r e c e i v e d p h e n e l z i n e s u l f a t e . C l i n e s c h m i d t , e t . a l . ( 1 0 , 11) s t u d i e d t h e in v i t r o b i o t r a n s f o r m a t i o n a s w e l l as t h e in v i v o metabolism of C"-phenelzine i n rats. Phenylaceti c a c i d w a s i d e n t i f i e d as t h e m a j o r m e t a b o l i t e . F i s c h e r , e t . al. ( 1 2 ) h a v e d e s c r i b e d t h e e f f e c t o f p h e n e l z i n e on t h e e x c r e t i o n of 6phenethylamine. 6.
Methods of A n a l y s i s 6.1
Spectrophotometric Analysis
6 . 1 1 F e i g l (13) has r e p o r t e d t h e reduct i o n of l i t h i u m and s o d i u m molybdophosphotungs t a t e s o l u t i o n by h y d r a z i n e d e r i v a t i v e s . T h i s r e a c t i o n h a s b e e n a d a p t e d t o t h e d e t e r m i n a t i o n of p h e n e l z i n e s u l f a t e r a w m a t e r i a l o r t a b l e t s by Bose and V i j a y v a r g i y a (14). R e d u c t i o n i s c a r r i e d o u t i n a l k a l i n e s o l u t i o n with t h e production of a d e e p b l u e c o l o r w h i c h is m e a s u r e d a t 6 5 0 nm.
6.12 A c o l o r i m e t r i c method b a s e d on t h e formation of t h e hydrazone w i t h v a n i l l i n has been d e v i s e d ( 1 5 ) . The r e a c t i o n i s c a r r i e d o u t i n 0.5 N m e t h a n o l i c h y d r o c h l o r i c a c i d . Absorbance o f t h e y e l l o w s o l u t i o n i s r e a d a t 4 0 0 nm. S i n c e p h e n e l z i n e d e g r a d e s by o x i d a t i v e d e s t r u c t i o n of t h e h y d r a z i n e f u n c t i o n , t h i s method i s s t a b i l i t y indicating
.
Polarography The h a l f wave p o t e n t i a l versus s t a n d a r d calomel e l e c t r o d e ) of t h e a c e t o n e d e r i v a t i v e of p h e n e l z i n e w a s r e p o r t e d ( 7 ) as - 1 . 3 3 V i n pH 5 . 9 p h o s p h a t e b u f f e r ( 0 . 1 M KH~POI, : N a 2 H P O 4 * 2 H 2 O ) employing a 0 . 0 1 % g e l a t i n s o l u t i o n as a maximum s u p p r e s s o r . The d i f f u s i o n c u r r e n t Four elecc o n s t a n t ( I d ) w a s approximately 2.5. t r o n s w e r e t r a n s f e r r e d f o r t h e r e d u c t i o n of t h e h y d r a z o n e d e r i v a t i v e . W a v e h e i g h t was p r o p o r t i o n a l t o c o n c e n t r a t i o n i n t h e r a n g e of 2 . 5 x lo-' t o 2.5 x M i n pH 5 . 9 p h o s p h a t e b u f f e r . 6.2
3Y.5
ROBERT E . D A L Y
6.3
Titrimetry P r o c e d u r e s b a s e d on t h e r e a c t i o n o f phenelzine s u l f a t e with iodine i n bicarbonate s o l u t i o n ( 2 ) and i n sodium h y d r o x i d e ( 1 6 ) and w i t h b r o m a t e and bromide i n a c i d s o l u t i o n ( 1 6 ) w i t h s u b s e q u e n t t i t r a t i o n of e x c e s s r e a g e n t ( i o d i n e o r bromide c o n v e r t e d t o i o d i n e ) i n a c i d s o l u t i o n w i t h t h i o s u l f a t e have been r e p o r t e d . B a s i c a l l y , t h e r e a c t i o n i n v o l v e d may be i l l u s t r a t e d a s follows:
Radecka and Nigam ( 1 7 ) have r e p o r t e d t h e d i r e c t t i t r a t i o n o f p h e n e l z i n e w i t h N-bromosuccinimide i n a c i d i c s o l u t i o n employing m e t h y l r e d a s t h e indicator. However, t h e i r r e s u l t s show a 5% p o s i t i v e b i a s . This b i a s is a t t r i b u t e d t o a l l y l i c b r o m i n a t i on by N -b r omosu c c i n i m i d e
.
6.4
Chromatography
Gas Chromatography C a r d i n i , e t . a l . (18, 1 9 ) and McMartin and S t r e e t ( 2 0 ) h a v e r e p o r t e d g a s chroma t o g r a p h i c methods which a r e r e p u t e d t o be quantitative. However, no q u a n t i t a t i v e d a t a a r e g i v e n . C o n d i t i o n s a r e r e p o r t e d i n T a b l e 11. 6.41
T h i n Layer Chromatography Q u a l i t a t i v e methods f o r t h e i d e n t i f i c a t i o n of p s y c h o t r o p i c a g e n t s have b e e n r e p o r t e d ( 2 1 , 2 2 , 2 3 , 2 4 , 2 5 ) . R f v a l u e s a r e summarized i n T a b l e 111. 6.42
6.5
Spot T e s t s A v a r i e t y of s p o t t e s t s have been re-
ported ( 2 6 ) .
T h e s e are summarized i n T a b l e 396
v.
T A B L E I1 - P a r a m e t e r s f o r G a s C h r o m a t o g r a p h y Ref. 18
19 w
c 4
20
Column g l a s s column 1 . 8 M x 2 mm.; 2 % GE-SE 3 0 on A e r o p a k 3 0 , 80100 m e s h
Column Temp.
Carrier Gas
program 70-250' a t 10.4O/
N2
Flow Rate
50 m l . / min.
Detector
FID
Detector Te m p
.
220'
C.
Injector Temp. 250'
C.
minute
Q
Stainless steel 140' c o l u m n 1 . 2 M. x 2 mm.; 15%c a r b o w a x 20 M o n C h r o m o s o r b W alkalinized w i t h 5% KOH
C.
Stainless steel 140' c o l u m n 6' x 1/8" ; 2 % SE 3 0 + 0 . 1 % t r i s t e a r i n on s i l a n i z e d acid w a s h e d C h r o m o sorb W
C.
He
40 m l . / min.
FID
250'
C.
I rn
z
rn
P z
rn
m C
r
n
>
2 N2
3 0 ml./ min.
FID
ROBERT
TABLE I11
-
E. D A L Y
S u p p o r t , S o l v e n t System and Rf
V a l u e s f o r TLC of P h e n e l z i n e S u l f a t e Reference
Support
Solvent System
Rf
(X
21
silica gel G
chloroform: m e t hano 1 (8:2)
21
silica gel G
chloroform: acetone: d i e t h y lamine ( 5 : 4: 1)
88
21
silica gel G
cyclohexane: chloroform: d ie t h y l a m ine ( 4 :5 :1)
62
22
silica gel G impregnated with 0 . 1 M N aOH
hexane:benzene: diethylamine ( 7 5 : 1 5 : 10)
51
22
silica gel G impregnated with 0.1 M N aOH
methanol
72
22
silica gel G imp re gn a t ed with 0 . 1 M NaOH
acetone
75
22
silica gel G impregnated with 0 . 1 M NaHSO
methanol
41
398
100
100)
PHENELZINE SULFATE
TABLE I11 f c o n t . 1
R e f er e n ce
Support
Solvent System
Rf
( x 100)
silica gel G impregnated with 0 . 1 M NaHSO
95% ethanol
25
silica gel G activated a t 110" C. f o r 30 m i n u t e s
chloroform: m ethano1
37
23
silica gel G a c t i v a t e d as above
chloroform: methanol : 2 0 % ammonium hydroxide ( 2 : 1:l)
56
24
silica gel G activated at 100' C . f o r 1 hour
methanol:12 N ammon i um hydroxide (100: 1.5)
74
24
silica gel G impregnated with 0 . 1 N NaOH and activated a s above
cyclohexane: diethylamine: benzene (75: 20: 15)
44
24
s i l i c a g e l G acetone impregnated and a c t i v a t e d as above
65
24
s i l i c a g e l G chloroform: impregnated methanol and a c t i v a t e d ( 9 0 : 10) as above
75
22
+ ,
23
(1:l)
399
ROBERT E. DALY
TABLE I11 ( c o n t . )
Reference
Support
Solvent System
Rf
(X
100)
24
silica gel G
benzene: ethanol: 1 2 N ammonium hydroxide ( 9 5 : 15: 5 )
50
25
silica gel G imp re g n a t e d with 0 . 1 N KHSO 4
95% e t h a n o l
49
25
silica gel G impregnated w i t h 0.1N
cyclohexane: benzene : d i et h y lamine ( 7 5 : 15: 10)
52
silica gel G imp r e g n a t e d with 0 . 1 N
methanol
72
silica gel G impregnated with 0.1 N
acetone
75
25
silica gel G imp r e g n a t e d with 0 . 1 N KHSO I,
methanol
62
25
silica gel G impregnated with 0 . 1 N NaOH
methyl acetate
53
N aOH
25
N aOH
25
N aOH
400
PHENE LZI NE SULFATE
TABLE I11 ( c o n t . ) Reference 25
Support
Solvent System
chromedi a n-butanol: (Whatman #41) 5% c i t r i c impregnated acid w i t h 5% s o d i u m d i h y d r og e n citrate (some K2HP04 a d d e d t o prevent tailing)
Rf (x 100) 51
ROBERT E. DALY
TABLE I V
-
Reagents and Reactions
F o r TLC P r o c e d u r e s Reagent
Reaction
21
Folin-Ciocalteau's Reagent
blue violet with heating
21
5% phosphomolybdic a c i d i n e t h a n o l f o l l o w e d by exposure of t h e p l a t e t o ammonia v a p o r
blue
22
1%i o d i n e i n m e t h a n o l
brown
23
50 m l . o f 0 . 2 % ninhydrin i n methanol + 1 0 m l . of g l a c i a l a c e t i c a c i d + 2 m l . of
r o s e orange
23
1 0 % F e r r i c c h l o r i d e ; 1%
fluorescent blue
23
S a t u r a t e d ammonium molybdate; s a t u r a t e d oxalic acid
celeste
24
Fo l i n - C i o c a l t e a u ' s Reagent
b l u i s h a t RT, b l u e when heated t o
Reference
2,4,6-trimethylpiperidine f l u o r e s c e i n : ammonia
100'
c.
24
Mandelin s R e a g e n t
pink a t RT, f l e s h a t 100' C. , f l u o r e s c e n t a f t e r heating
24
C i nn ama l d e h y d e R e a g e n t 5 m l . cinnamaldehyde + 95 m l . ethanol + 5 m l . conc. HC1
y e l l o w a t RT
402
PHENELZINE SULFATE
TABLE I V ( c o n t . )
Reference
Reagent
Reaction
24
0.25 g . p-dimethylaminobenzaldehyde + 5 g . of 85% p h o s p h o r i c a c i d + 20 m l . of water
lemon chromo after heating t o 1 0 0 ' C. f o r 1 0 min. and t h e n exposed t o ammonia v a p o r
24
Furf u r a l r e a g e n t s o l ' n A.) 1 m l . f u r f u r a l i n 99 m l . of acetone s o l ' n B . ) 4 m l . of c o n c . H 2 S 0 4 i n 96 m l . o f a c e t o n e
brown a t 1 0 0 '
25
P l a t e examined u n d e r s h o r t w a v e W l i g h t and t h e n s p r a y e d w i t h t h e following sequence of r e a g e n t s :
C.
iodine
brown
Dragendorff ' s
orange
sodium n i t r i t e
d e e p brown
i odop l a t in a t e until a color develops
variously colored
403
ROBERT E. DALY
TABLE V
-
Spot Tests f o r Phenelzine S u l f a t e
Reagent
Response 0 . 5 mg.
2 0 1-14.
Froehde ' s Reagent
blue* It. b l u e
blue
Mandelin ' s R e a g e n t
green blue* brown
green faint orange
Marquis ' Reagent Mecke's R e a g e n t
r e d orange* grey
pink
R e i c k a r d ' s Reagent
It. blue
It. b l u e
F l u e c k i g e r ' s Reagent
brown
V i t a l i ' s Reagent
a. 1 b.)
Was i c k y ' s R e agen t
red o r a n g e
*
=
a f t e r warming
404
- -blue orange
---
--- --
PH E N E LZ IN E SULFATE
TABLE V I
- Composition o f Spot T e s t Reagents
Reagent
Composition
Froehde I s Reagent
0 . 5 % a q u e o u s ammonium m o 1y b d a t e
Mande l i n I s R e a g e n t
0 . 5 % a q u e o u s ammonium vanadate
Marquis
1 p a r t 37% formaldehyde i n 20 p a r t s concentrated
Reagent
sulfuric acid, freshly prepared before use Mecke ' s R e a g e n t
0 . 5 % aqueous s e l e n i o u s a c i d
Reickard I s Reagent
1%a q u e o u s sodium t u n g s t a t e
F l u e c k i g e r ' s Reagent
0 . 5 % s o l u t i o n of t i t a n i u m d i o x i d e ( T i 0 2 1 i n concentrated sulfuric acid
V i t a l i s Reagent
a.) b.)
Wasicky s R e a g e n t
(H2SeO 3 1
fuming n i t r i c a c i d
3% potassium hydroxide
i n ethanol
1 0 % s o l u t i o n o f p-dimethylaminobenzaldehyde i n g l a c i a l acetic acid
305
ROBERT E. DALY
7.
References R a j e s w a r a n a n d P . L. K i r k , B u l l . N a r c o t i c s , U . N . , D e p t . S o : L 4 i l ~ f f ~ i 1r 4 s, No. I, 1 9 ( 1 9 6 2 ) .
1.
P.
2. 3. 4.
U.S.P. X V I I I . J . H . B i e l , US 3 , 0 0 0 , 9 0 3 , S e p t . 1 9 , 1 9 6 1 . A. S a c h a a n d L . M a l a s n i c k i , P o l 4 6 , 2 3 7 , O c t . 3 0 , 1 9 6 2 , C . A . 60, 4 5 4 f ( 1 9 6 3 ) . J . H . B i e l , A. E . D r u k k e r , T . H . M i t c h e l l , E . P . S p r e n g e l e r , P . A . N u h f e r , A. C . Conway and A. H o r i t a , J . A m . Chem. S o c . 8 2 , 2805 ( 1 9 5 9 ) . E . V o t o c e k a n d 0. L e m i n g e r , ColI. C z e c h . Chem. Commun. 4 , 2 7 1 ( 1 9 3 2 ) , C. A . 2 6 ,
5.
6. 7. 8. 9.
10.
11. 12.
13. 14.
15. 16. 17. 18. 19. 20.
5294 ( 1 9 3 2 ) . L . S c h l i t t , M. R i n k a n d M. von S t a c k e l b e r g , J , E Z e c t r o a n a l . Chem. 2 3 , 10 (1967). L . E . E b e r s o n a n d K . P e r s s o n , ,J. Med. Pharm. Chem. 5, 7 3 8 ( 1 9 6 2 ) . J . D r a b n e r a n d W . S c h w e r d , F r e s e n i u s ' Z. A n a l . Chem. 2 4 3 , 9 2 ( 1 9 6 8 ) . C. A . 70, 558941 ( 1 9 6 9 ) B . Van C l i n e s c h m i d t a n d A. H o r i t a , Biochem. Pharmacol. 1 8 , 1 0 1 1 (1969) Idem., i b i d . , 1021 (1969). E . F i s c h e r , B . H e l l e r , a n d A . H . Miro, A r z n e i m i t t e l - F o r s c h . 18, 1 4 8 6 ( 1 9 6 8 ) . F. F e i g l , "Spot T e s t i n O r g a n i c Analysis", E l s e v i e r Company, L o n d o n , 1 9 5 6 , p . 1 2 8 . B . C . Bose a n d R . V i j a y v a r g i y a , I n d i a n J . Pharm. 2 8 , 328 ( 1 9 6 6 ) . R. E . D a l y , u n p u b l i s h e d r e s u l t s M. M a r z a d r o a n d A. D e C a r o l i s , R e n d . I s t . S u n i t a 2 6 , 629 ( 1 9 6 3 ) . C . R a d e c k a a n d I . C. Nigam, Can. J . P h m m S c i . 2 , 17 (1966). C . C a r d i n i , V . Q u e r c i a a n d A . C a l o , :J. C h r o m a t o g . 37, 1 9 0 ( 1 9 6 8 ) . U. Q u e r c i a , C . C a r d i n i a n d A . Calo, i i ( ' l . C h i m . Farm. 1 0 7 , 3 8 3 ( 1 9 6 8 ) . C. M c M a r t i n a n d H . V . S t r e e t , J . Chromatog. 2 2 , 274 ( 1 9 6 6 ) .
.
.
406
PHENELZINE SULFATE
7.
References
21. 22. 23. 24. 25. 26.
(cont. )
E. S c h m i d , E. Hoppe, C . M e y t h a l e r a n d L.
Z i c h a , A r z n e i m i t t e Z - F o r s c h , 1 3 , 9 6 9 (1963). W . W. F i k e , A n a l . Chem. 38, 1 6 9 7 ( 1 9 6 6 ) . A . A l e s s a n d r o , F. Mari a n d S . S e t t e c a s e , F arma co , Ed. Prat. 22, 437 ( 1 9 6 7 ) . I. Z i n g a l e s , J . C h r o m a t o g . 3 1 , 405 (1967). I. S u n s h i n e , W. W . F i k e a n d H . L a n d e s m a n , J . F o r e n s i c S c i . 1 1 , 428 ( 1 9 6 6 ) . P. R a j e s w a r a n a n d P . L . K i r k , B u l l . Narcotics, U.N., Dept. S o c i a l A f f a i r s 13, No. 3, 15 ( 1 9 6 1 ) .
407