ACETOHEXAMIDE

Abdullah A. Al-Badr and Humeida A. El-Obeid

Pharmaceutical Chemistry Department

College of Pharmacy
King Saud University

Riyadh, Saudi Arabia

ANALYTICAL PROFILES OF DRUG SUBSTANCES

AND EXCIPIENTS VOLUME 21

Copyright 0 1992 by Academic Press, Inc.

All rights of reproduction reserved in any form

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

C O N T E N T S
1.

DESCRIPTION
1 . 1 Nomenclature
1.1.1
Chemical Names
1.1.2
Genermic Names
1.1.3
Trade Names
1 . 2 Formulae
1.2.1
Empirical
1.2.2
Structural
1.2.3
GAS No.
1.3 Molecular Weight
1 . 4 Elemental Composition
1 . 5 Appearance

2.

PHYSICOCHEMICAL PROPERTIES
2 . 1 Melting Range
2.2 S o l u b i l i t y
2 . 3 Polymorphism
2 . 4 Thermal Analysis
2 . 5 X-ray Powder D i f f r a c t i o n
2.6 Spectral Properties
2.6.1
U l t r a v i o l e t Spectrum
2.6.2
I n f r a r e d Spectrum
2.6.3
Proton Nuclear Magnetic Resonance (PMR)
Spectrum
2.6.4
lac-Nuclear Magnetic Resonance (SC-NMR)
Spectrum
2.6.5
Mass Spectra

3.

SYNTHESIS

4.

METHODS OF ANALYSIS
4 . 1 T i t r i m e t r i c Methods
4.1.1
Nonaqueous
4.1.2
Gravimetric
4.1.3
Campleximetric
4.2 Spectrometric Methods
4.2.1
Colorimetric
4.2.2
U1t r a v i o l e t
4.2.3
Infrared
4.2.4
Fluormetric
4.2.5
Proton Magnetic Resonance

ACETOHEXAMIDE

4.3

5.

Chromatographic Methods
4.3.1
Thin-Layer Chromatography (TLC)
4.3.2
Gas-Liquid Chromatography (GLC)
4.3.3
High-Performance Liquid Chromatography
(HPLC)

PHARMACOKINETICS
5 . 1 Introduction
5.2 Mechanism o f Action
5.3 Onset and Duration o f Action
5.4 Absorption
5.5 Distribution
5.6 Metabol ism
5.7 Excretion
5.8 Half-Life

ACKNOWLEDGEMENT

REFERENCES

ABDULLAH A. AL-BADR AND HUMEIDA A . EL-OBEID

ACETOHEXAMIDE
1.

DESCRIPTION

1 1 Nomenclature
1.1.1

Chemical Names

4-Acetyl-N-[(cyclohexylamino)carbonyl]benzenesul-

fonamide

l-[(pAcetylphenyl)sulfonyl]-3-cyclohexylurea.
3-Cyclohexyl-l-(pacetylphenylsulfonyl)urea.

N-(pAcetyl benzyl sul fonyl l-N -cyclohexyl urea.

1.1.2

Generic Names

Acetohexamide, Acetohexamidum

1 1.3

Trade Names

Cycl am1de , Dime 1in , Dime1o r , Dyme 1o r , Gamadiabet,

Metaglucina, Ordimel, Tsiklamid.
1.2

Formulae
1.2.1.

EmDlriCal

Ct sHzoNz04S

1.2.2

Structural

1.2.3

CAS No.

[968-81-01
1.3

Molecular Weight
324.42 (1)

1.4

Elemental ComDosltion
C 55.54%, H 6.21%, N 8.64%, 0 19.73%,

S 9.89% (1).

ACETOHEXAMIDE

1.5

Armearance

A w h i t e , c r y s t a l l i n e powder; o d o r l e s s o r almost
odorless (2).

2.

PHYSICOCHEMICAL PROPERTIES
2.1

M e l t i n g Range

C r y s t a l s from 90% aqueous e t h a n o l m e l t between

188-190" ( 3 ) . Crystals from d i l u t e ethanol m e l t between
175-177 (4).
2.2

Solubility

Soluble i n p y r i d i n e , s l i g h t l y soluble i n alcohol

and chloroform. I n s o l u b l e i n water and ether ( 1 ) .
2.3

P01YmOrDh'ism

The l i t e r a t u r e r e p o r t s i n d i c a t e t h a t acetohexamide
e x i s t s as more t h a n one polymorphic forms ( 5 - 1 5 ) .
G i rgis-Takla and Chroneos (5) prepared acetohexamide
polymorphs A and B by h e a t i n g t h e drug ( 1 gm) w i t h
g l a c i a l a c e t i c a c i d o r chloroform respectively, before
c r y s t a l l i z a t i o n a t 1 0 5 ' and room t e m p e r a t u r e
respectively. While acetohexamide polymorph A showed a
m e l t i n g range o f 180"-183', t h e acetohexamide polymorph
B melted a t 183'-185".
D i f f e r e n t i a l scanning
calorimetry and I R spectroscopy showed t h a t c r y s t a l s o f
polymorph B were converted t o polymorph A by grinding.
A c c o r d i n g l y , t h e s e r e s u l t s i n d i c a t e t h a t any
i d e n t i f i c a t i o n t e s t u t i l i z i n g g r i n d i n g may f a i l to
i d e n t i f y t h e two polymorphs. I n t h e i r phystco-chemical
studies on t h e polymorphism o f acetohexamide, Kuroda e t
a7 (6) obtained t h r e e polymorphs o f acetohexamide by
r e c r y s t a l l i z a t i o n from d i f f e r e n t solvents. These are
f o r m I,f o r m I 1 and CHC13-11. A l t h o u g h t h e X-ray
d i f f r a c t i o n p a t t e r n s , I R s p e c t r a and d i f f e r e n t i a l
scanning calorimeter curves o f t h e CHC13-I1 polymorph
were i d e n t i c a l w i t h those o f polymorph 11, t h e CHC13-I1
t y p e c o n t a i n e d a C H C l j molecule which c o u l d n o t be
removed by normal d r y i n g condition. Polymorph CHC13-I1
seemed t o be unsuitable f o r medicinal use. Form I 1 i s
1.2 times more soluble than form I.

ABDULLAH A. AL-BADR AND HUMElDA A. EL-OBEID

Burger ( 7 ) c h a r a c t e r i z e d t h e t h r e e p o l y m o r p h i c
m o d i f i c a t i o n s o f acetohexamide by thermomicroscopy,
d i f f e r e n t i a l scanning calorimetry and I R spectroscopy.
The s o l u b i l i t y behavior o f the three modifications o f
the drug i n butanol and buffer solutions i s described
and d i s c u s s e d i n r e l a t i o n t o thermodynamics and
pharmacological parameters such as b i o a v a i l a b i l i t y from
t a b l e t s and USP X I X d i s s o c i a t i o n t e s t . M u e l l e r and
L a g a s ( 8 ) h a v e c o n f i r m e d t h e e x i s t e n c e and
characterized two polymorphic forms o f acetohexamide
using d i f f e r e n t i a l scanning calorimetry, thermogravimetric analysis, scanning e l e c t r o n microscopy as we1 1
as I R , NMR and X-ray analysis. The study has pointed t o
the u n s u i t a b i l i t y o f phosphate b u f f e r s o l u t i o n which i s
sometimes prescribed f o r use i n the d i s s o l u t i o n t e s t s
o f the drug since the s a l t o f the drug c r y s t a l l i z e s out
during the t e s t . I n another study (9) the same authors
reported t h a t form Idecomposed during melting and form
I1 melted a t 180" and then r e c r y s t a l l i z e d t o form I.A t
a heating r a t e o f lO'/minute melting points o f 193.6"
and 180.5" were found f o r forms Iand 11, respectively.
No morphological differences were observed between the
two forms. I n s o l u b i l i t y and d i s s o l u t i o n r a t e studies
i n sodium potassium b u f f e r , potassium acetohexamide
c r y s t a l l i z e d e x h i b i t i n g a lower s o l u b i l i t y than
acetohexamide. I n t h i s respect, form I 1 was transferred
t o potassium acetohexamide more quickly than form I.
Yokoyama e t a7 (10) calculated the thermodynamic values
o f forms I and I 1 o f acetohexamide from s o l u b i l i t y
measurements. The t r a n s i t l o n temperature and the heat
o f t r a n s i t i o n were 154" and 230 cal/mole, respectively.
I t i s found t h a t the polymorphic forms o f acetohexamide
d i d n o t a f f e c t i t s b i o a v a i l a b i l i t y when i n v i v o
absorption studies o f form I & I 1 were c a r r i e d out i n
b e a g l e dogs. The p r e p a r a t i o n o f f o u r c r y s t a l l i n e
modifications o f acetohexamide was reported (11). Their
thermograms, I R s p e c t r a , X-ray d i f f r a c t i o n and
s o l u b i l i t y are also reported. Two o f the forms reverted
t o the most stable form on storage i n solution.
S o l i d dispersion o f acetohexamide was studied by Graf
e t a7 (12-14) u s i n g d i f f e r e n t polymers and v a r i o u s
ratios.
C o p r e c i p i t a t e s o f acetohexamide w i t h
polyethylene g l y c o l (PEG 6000) were prepared by t h e
s o l v e n t method w i t h ethanol ( c r y s t a l l i n e form I)
or
with chloroform ( c r y s t a l 1ine form 111). Phase diagrams

ACETOHEXAMIDE

o f form I-PEG and form 111-PEG coprecipitates were o f

the p e r i t e c t i c type and the molecular compounds were
formed i n the r a t i o o f 1 mole o f acetohexamide t o 4
moles o f PEG. The e u t e c t l c t e m p e r a t u r e , e u t e c t i c
composition and the end o f melting o f the two binary
system were, however, d i f f e r e n t ( 1 2 ) .
Both the
s o l u b i l i t y and t h e s o l u t i o n r a t e were increased by PEG.
S i m i l a r r e s u l t s were o b t a i n e d by s u b s t i t u t i n g
p o l y ( v i n y l p y r r o 1 i d o n e ) (PVP) f o r PEG ( 1 3 ) . Also,
c o p r e c i p i t a t e s o f acetohexamide-PVP ( i n e t h a n o l )
containing drug concentrations o f 60% o r more showed
the same X-ray d i f f r a c t i o n pattern as t h a t o f form I.
Increasing the PVP concentration ( > 55%) d i d n o t show
any c r y s t a l behavior i n the X-ray analysis. I n another
r e p o r t Graf
e t a7 ( 1 4 ) d e s c r i b e d t h e methods o f
p r e p a r a t i o n and t h e e f f e c t o f t h e s o l v e n t s on t h e
acetohexamide-PVP coprecipi tates. They were obtained
from ethanol o r chloroform by evaporating the solvent
a t room temperature, under vacuum or by spray drying.
Changing t h e s o l v e n t and/or i t s e v a p o r a t i o n r a t e
affected the polymorphic form, the c r y s t a l l i n i t y and
the s o l u t i o n r a t e o f acetohexamide i n coprecipitates
containing less than 70% PVP.
Kassem e t a7 (15) studied the enhancement o f the r a t e
o f release o f acetohexamide from i t s t a b l e t s by t h e
f o r m a t i o n o f s o l i d d i s p e r s i o n s w i t h each o f f o u r
water-sol uble pol ymers prepared in d i f f e r e n t r a t i0s.
The polymers were r a t e d i n t h e o r d e r o f decreasing
r e l e a s e r a t e s as f o l l o w s : PEG 6 0 0 0 ,
PVP,
hydroxypropylmethylcellulose, methylcellulose.
2.4

Thermal Analysis

The h e a t o f f u s i o n and m e l t i n g p o i n t o f
acetohexamide were done u s i n g DuPont TA 9900 on t h e
DSC- u n i t a t a temperature range i n d i c a t e d i n t h e
thermogram (Figure 1). Sample i s done i n duplicate and
the average o f t h e value i s reported as follows:

AHf
2.5

63.7 kJ/mOle

Purity

99.82%

Tm

187.45 C

X-ray Powder D i f f r a c t i o n

The X-ray powder d i f f r a c t i o n p a t t e r n o f acetohexamide was determined using P h i l i p s f u l l automated X-ray

d i f f r a c t i o n spectrogoniometer equipped w i t h PW1730/10

PURITY v l . l A
F i g u r e 1. Thermal cu rve o f acetohexamide.

ACETOHEXAMIDE

J
Figure 2 . X-Ray powder d i f f r a c t i o n p a t t e r n of acetohexamide.

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

10

generator. Radiation was provided by a copper t a r g e t

(Cu anode 2000W, X = 1.5480 A), high I n t e n s i t y X-ray
tube operated a t 40 kV and 35 mA. The monochromator was
a curved s i n g l e c r y s t a l one (PW1752/00). Divergance
s l i t and t h e r e c e i v i n g s l i t were 1 and 0.1
r e s p e c t i v e l y . The scanning speed o f t h e gonlometer
(PW1050/81) used was 0.02 2 8 p e r second. The
instrument i s combined w i t h P h i l i p s PM8210 p r i n t i n g
r e c o r d e r w i t h b o t h analogue r e c o r d e r and d i g i t a l
p r i n t e r . The goniometer was a l i g n e d using s i l i c o n
sample before use.
The X-ray pattern o f acetohexamlde I s presented i n
Figure 2. The interplanar distance d(A) and r e l a t i v e
i n t e n s i t i e s 1/10 are shown i n Table 1.
2.6

Spectral ProDerties
2.6.1

U l t r a v i o l e t Spectrum

The u l t r a v i o l e t a b s o r p t i o n s p e c t r u m o f
acetohexamide i n methanol was obtained on a Cary 219
spectrophotometer. The spectrum, shown i n Figure 3, i s
characterized by two maxima. The one w i t h a Xmax a t 247
nm i s t y p i c a l o f s u b s t i t u t e d acetophenones. The
absorption a t X m a x 283 nm represents a conjugated
aromatic r i n g system.
2.6.2

I n f r a r e d SDectrum

The i n f r a r e d absorption spectrum o f acetohexamide,

obtained from a potassium bromide d i s p e r s i o n , was
recorded on a Pye Unicam SP 1025 spectrometer and i s
shown i n Figure 4. The assignment o f the c h a r a c t e r i s t l c
bands are shown i n Table 2.

220

XO

nm

300

3 50

400

450

Figure 3 . U l t r a v i o l e t spectrum o f acetohexamide in methanol.

v,.
m.

c
C
N.

I--

6,.
al.
Q.

t-.

v),

*I

cn

-I-

m
Y
"

E
N

z!
Y
W

c
0
c,
W

tcl

'*0

f
V
W

L
c,
0

cn

fu
I

13

ACETOHEXAMIDE

Table 1: X-ray d i f f r a c t i o n p a t t e r n o f acetohexamfde

d
1/10

d(A)

1/10

d(A)

15.74
9.47
7.85
7.21
5.30
4.99
4.93
4.55
4.30
4.19
4.08
3.92
3.78
3.60
3.50
3.28
3.26
3.15
3.07
3.01
2.91
2.88
2.74
2.65
2.61
2.58

31.25
30.04
6.89
2.25
100.00
8.28
10.95
4.71
5.30
15.19
23.07
5.44
2.82
24.35
4.52
23.29
9.83
5.72
9.36
1.26
7.99
2.79
4.08
1.51
2.15
1.80

2.55
2.49
2.40
2.36
2.31
2.29
2.27
2.24
2.18
2.15
2.13
2.09
2.04
1.99
1.95
1.94
1.89
1.81
1.77
1.72
1.66
1.64
1.61
1.57
1.47
1.35

= Interplanner distance

1/10
1.82
1.75
6.19
4.44
5.26
2.20
1.81
2.54
2.04
2.38
1.20
2.56
4.51
1.69
2.91
4.16
1.50
1.48
1.29
1.77
1 .oo
1.18
1.16
1.32
0.85
0.80

r e l a t i v e i n t e n s i t y (based on highest i n t e n s i t y of
100).

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

14

Table 2:

I n f r a r e d c h a r a c t e r i s t i c bands and t h e i r
assignments.

Frequency (cm-

Assignment

3340, 3270

Amide N-H s t r e t c h

2980, 2940

Aromatic C-H s t r e t c h

1710, 1680

Conjugated

1602 , 1600

Aromatic C s t r e t c h

1455

E-

CH3 bending

1345
780, 760

2.6.3

Aromatic C-H out o f plane

bending

Proton Nuclear Magnetic Resonance ( W R l

Spectrum

Acetohexamide s o l u t i o n i n DMSO-de was used t o

obtain the PMR spectrum on a Varian XL 200 MHr FT NWR
spectrometer u s i n g TMS as i n t e r n a l reference. The
spectrum i s shown i n Figure 5. The number o f protons i s
established by both i n t e g r a t i o n o f the area under the
curve and t h e m u l t i p l i c i t i e s o f t h e peaks. Table 3
a s s i g n s t h e chemical s h i f t s t o t h e i r r e s p e c t i v e
protons. F u r t h e r evidence f o r p r o t o n assignment i s
obtained from the HETCOR pulse sequence (Figure 9).

Figure 5. PMR spectrum of acetohexamide i n DMSO-dG

as internal reference.

using TMS

F i g u r e 6. 1 3 C NMR spectrum o f acetohexzmide i n DMSO-ds

TMS as internal reference.

using

Figure 7.

1 3 C NMR spectrum o f acetohexarnide u s i n g DEPT

ex P e r iment

F i g u r e 8.

1 3 C I M R sDectrum of acetohexanide u s i n g APT

expe r irnent .

Figure 9.

13C N M R spectrum of acetohexamide using HETCOR

experiment.

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

20

Table 3:

Assignment o f the NMR chemical s h i f t s t o the

d i f f e r e n t protons

Chemical s h i f t
(6)

Multiplicity

Proton
assignment

No. o f
protons

1.09 - 1.71

mu1ti p l e t

Cyclohexyl
ring3

11

8.06

2.66

singlet

CH3-0

6.45

doublet

CH-NH

8.19

mu1t ip l e t

Aromat ic d

2.6.4

13C-Nuclear Magnetic Resonance

SDect rum

(13C

NHR)

The 1 3 C NMR spectra o f acetohexamide i n DMSO-ds

using TMS as i n t e r n a l reference are obtained using a
V a r i a n XL 200 MHz p u l s e FT s p e c t r o m e t e r and a r e
p r e s e n t e d i n F i g u r e s 6-9.
The assignment o f t h e
chemical s h i f t s and the degree o f carbon protonation,
presented i n Table 4, are achieved u s i n g t h e DEPT
(Figure 7) and APT (Figure 8) experiments as well as
t h e HETCOR p u l s e sequence ( F i g u r e 9 ) and t h e
approximate a d d i t i v e e f f e c t s o f substituents.
2.6.5

Mass SDectra

The 70 eV e l e c t r o n impact mass spectrum o f

acetohexamide, presented i n Figure 10, was obtained on
Varian MAT 311 mass spectrometer u s i n g i o n source
pressure o f 10-0 Torr, i o n source temperature o f 180'C
and an emission current o f 300 M. The molecular i o n i s
detectable a t m/e 324 and the base peak a t m/e 56. A
proposed fragmentation p a t t e r n and t h e mass/charge
r a t i o s o f the major fragments are shown I n Scheme 1.

21

ACETOHEXAMIDE

Table 4:

Assignment o f the carbon chemical s h i f t s .

Chemical s h i f t
(PHI

Carbon assignment

Number o f Protons
attached

24.26

25.07

26.99

32.33

48.30

127.73

128.73

140.00

143.93

150.45

197.30

The chemical i o n i z a t i o n spectrum, shown i n Figure 11,

was obtained on Finnigan 4000 mass spectrometer using
methane gas as a reagent with ion electron energy o f
100 eV, ion source pressure o f 0 . 3 T o r r , i o n source
temperature o f 150C and emission current o f 300 pA.
The spectrum i s dominated by a quasimolecular i o n (M +
1 ) . Two peaks appearing a t m/e 353 and m/e 365 are
a t t r i b u t a b l e t o the t r a n s f e r o f carbocations from the
c a r r i e r gas. The mass s p e c t r a l assignment o f t h e

N
N

F i g u r e 10. Electron impact mass spectrum o f

acetohexamide.

Figure 11. Chemlcal ionization mass spectrum o f

acetohexarnide.

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

24

prominent ions under the chemical i o n i z a t i o n conditions

are presented i n Table 5.
Table 5:

3.

Mass spectral assignment o f acetohexamide

using chemical ionization.

M/e

Species

365

[M

C3H5]+

353

[M

CzHs]+

325

[M

H (MH)1+

324

[MI+

SYNTHESIS

Marshall e t a7 ( 4 ) reported a method o f synthesis

o f acetohexamide which i n v o l v e s t h e r e a c t i o n o f t h e
diazonium s a l t from paminoacetophenone w i t h s u l f u r
dioxide t o a f f o r d the sulfonyl chloride which i s then
converted t o the sulfonamide by reaction w i t h a m n i a .
E l a b o r a t i o n v i a t h e carbamate w i t h cyclohexylamine
a f f o r d s acetohexamide. Another r e p o r t e d method ( 1 6 )
uses p-chloroacetophenone as t h e s t a r t i n g m a t e r i a l .
Both methods are o u t l i n e d i n Scheme 2.

25

ACETOHEXAMIDE

Scheme 1: Proposed mass fragmentation pattern o f

acetohexamide

n
0

W-QO

mle 3 2 4

0-H
0
m/e 243

+
NH
I1

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

26

Scheme 1 Continued ...

mle183

I -CH,-CO

m l e 324

m /el41

mle 324

m l e 200

- YN0,S

0
C H3I;

mle76

m l e 104

mle 119

-i

21

ACETOHEXAMIDE

Scheme 1 Continued

...

m/e 324

1
O N H I +

II

2 68

28

ABDULLAH A . AL-BADR A N D HUMEIDA A. EL-OBEID

Scheme 3: Synthesis o f acetohexamide

Method 1 (4)

SO,-NH-C-NH

Method 2 (16)

0
CH,t@

c H 3 - ! G so, CI

0
S03Na p0c13*

SO,NH,-

ACETOHEXAMIDE

4.

29

METHODS OF ANALYSIS

4.1

T i t r i m e t r i c Methods

4.1.1

Nonaaueous

A non-aqueous t i t r a t i o n method f o r t h e d r u g and

other hypoglycemic and d i u r e t i c agents was reported by
Agarwal and Walash (17). The drug i n t a b l e t o r pure
form was d i s s o l v e d i n t e t r a m e t h y l urea and t i t r a t e d
w i t h 0.1 N l i t h i u m m e t h o x i d e i n benzene-methanol
medium. The end p o i n t was determined u s i n g 0.2% azo
v i o l e t i n toluene as i n d i c a t o r . Recovery ranged from
98.8% t o 101.6%.

A n o t h e r non-aqueous t i t r a t i o n p r o c e d u r e , f o r t h e
q u a n t i t a t i v e a n a l y s i s o f t h e d r u g and o t h e r
h y p o g l y c e m i c s u l f o n y l u r e a s u s i n g HC104 t i t r a t i o n
method, was a l s o reported (18).

4.1.2

Gravimetric

Amer and Walash (19, 20a) r e p o r t e d a method f o r

t h e g r a v i m e t r i c d e t e r m i n a t i o n o f acetohexamide by
treatment w i t h 2,4-dinitrophenylhydrazine t o
p r e c i p i t a t e t h e h y d r a z o n e (19). A m i x t u r e o f
acetohexamide, tolbutamide and chlorpropamide was a l s o
determined g r a v i m e t r i c a l l y (20a).

4.1.3

Compleximetric

G u e r e l l o and Dobrecky (21) have d e s c r i b e d a

procedure f o r t h e compleximetric e v a l u a t i o n o f
medications w i t h hyoglycemic a c t i o n i n c l u d i n g
acetohexamide. The procedure permits the determination
o f t h e hypoglycemic sulphonylureas. A weighed amount o f
drug was h y d r o l y s e d by h e a t i n g f o r 30 minutes w i t h
d i l u t e aqueous sodium h y d r o x i d e and t h e s o l u t i o n
n e u t r a l i z e d w i t h 0.1 N HC1, t r e a t e d w i t h 0.1 M CuSO4,
then w i t h b u f f e r s o l u t i o n t o pH 6, and f i l t e r e d . The
excess C U + ~ i n t h e f i l t r a t e was d e t e r m i n e d by
complexometric t i t r a t i o n w i t h 0.02 M EDTA disodium s a l t
u s i n g 1-(2-pyridylazo)-2-naphthol
as i n d i c a t o r . The
method i s applicable t o evaluate drugs i n t a b l e t .

ARDUI.1.AH A. AL-BADR AND HUMEIDA A. EL-OBEID

30

4.2

SDect romet r i c
4.2.1

Colorimetric

Reaction o f acetohexamide w i t h 2,4-dinitrophenylhydrazine t o produce t h e colored hydrazone was used by

Amer and Walash (19) t o determine t h e drug c o l o r i m e t r i c a l l y . The c o l o r e d p r o d u c t was d i s s o l v e d i n KOH and
determined a t 480 nm. The accuracy o f the method was
claimed t o be 100%. A n i n h y d r i n c o l o r i m e t r i c method f o r
some o r a l hypoglycemic agents was a l s o reported (20b).
Meier e t a7 ( 2 2 ) analysed acetohexamide and o t h e r
h y p o g l y c e m i c a g e n t s by d i s s o l v i n g t h e d r u g i n
chloroform, adding calcium acetate (1% i n methanol),
propylamine (5% i n methanol), d i l u t i n g w i t h chloroform
and reading the absorbance a t 565 nm a f t e r 15 minutes.
Pharmaceutical preparations may be estimated s i m i l a r l y .
4.2.2

U l t r a v i o l e t (UV)

Solomonova and D v o r n i t s k a y a ( 2 3 ) d e t e r m i n e d
acetohexamide by measuring t h e absorbance a t 229 nm i n
ethanol or 0.1 M sodium hydroxide. Other UV t e s t s f o r
t h e drug are a l s o reported (24, 25).
4.2.3

Infrared (IR)

Acetohexamide and o t h e r s u l p h o n y l u r e a s were

analysed by IR (22). A t e s t have a l s o been described
(24).
Lazaryan (26) determined t h e d r u g and o t h e r
h y p o g l y c e m i c a g e n t s by i n f r a - r e d a b s o r p t i o m e t r i c
determination. A sample i s t r e a t e d with chloroform and
t h e s o l u t i o n from t h e t a b l e t sample i s f i l t e r e d . A
p o r t i o n o f s o l u t i o n i s d i l u t e d w i t h chloroform and t h e
absorbance i s measured a t 1722 t o 1715 cm-1 i n 0.25 m
NaCl c e l l against chloroform.
4.2.4

F1uoromet r ic

G i r g i s - T a k l a and Chroneos ( 2 7 ) d e s c r i b e d a
s e n s i t i v e method f o r t h e f l u o r o m e t r i c determination o f
t h e d r u g i n plasma o r i n t a b l e t s by means of i t s
r e a c t i o n w i t h 1 - m e t h y l n l c o t l n a m i d e . The l i m i t o f
d e t e c t i o n was approximately 0.2 Mg o f t h e drug/mL and
t h e r e l a t i v e standard d e v i a t i o n was 31% f o r 2 Ng/ml i n

ACETOHEXAMIDE

31

plasma. The method i s s u i t a b l e f o r plasma samples

containing 0.5-2.5 Mg o f the drug/ml.
4.2.5

Proton Magnetic Resonance

Al-Badr and Ibrahim (28) described a simple, r a p i d

and accurate method f o r t h e assay o f the drug and other
hypoglycemic agents u s i n g p r o t o n magnetic resonance
spectrometry. The pure drug o r i n t a b l e t form, can be
determined using DMSO-ds as solvent and maleic a c i d as
i n t e r n a l standard.
The reported recovery i s 100 f 1.5% f o r pure drug and
98 t o 99.6 f 1.4% f o r t a b l e t s .
4.3

ChromatonraDhic Methods
4.3.1

Thin-Layer ChromatonraDhY (TLC]

Gergis-Takla and Josh1 (29) reported a TLC method

f o r the i d e n t i f i c a t i o n , assay and p u r i t y determination
o f t h e drug and other hypoglycemic agents i n powder and
i n t a b l e t f o r m u l a t i o n . The d r u g was d e t e c t e d by
d i s s o l v i n g powdered t a b l e t s o r powder i n
dichloromethane-acetone m i x t u r e (2: 1) and chromatographing t h e s o l u t i o n on s i l i c a gel F 2 5 4 p l a t e s with
cyclohexane-chloroform-acetic a c i d and e t h a n o l
(10:7:2:1).
For q u a n t i t a t i v e determination, t h e spots
were s e p a r a t e d , e l u t e d w i t h m e t h a n o l i c sodium
hydroxide,
d i l u t e d w i t h m e t h a n o l i c HC1 and t h e
absorbance was measured.
Surborg and Roeder (30) have recommended c o n s t a n t b o i l i n g s o l v e n t m i x t u r e s f o r t h e development o f
chromatograms on s i l i c a gel f o r acetohexamide and o t h e r
a n t i d i a b e t i c drugs: propanol-cyclohexane (37:163),
propanol-benzene-cyclohexene (9:14:27), and cyclohexane
- isopropanol (177:23). The R f values o f t h e drugs a r e
t a b u l a t e d , s p o t s were l o c a t e d by v i e w i n g i n 254 nm
radiation.
4.3.2

Gas L i a u i d ChromatonraDhY (GLC)

Kleber e t a l . (31) determined acetohexamide and

hydroxyhexamide i n b i o l o g i c a l f l u i d s u s i n g GLC.
Tolbutamide was used as an i n t e r n a l standard and M-HC1
was added t o the sample o f plasma o r urine, the m i x t u r e

32

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

was shaken w i t h t o l u e n e and was c e n t r i f u g e d . The

separated organic phase was shaken w i t h 7.5% KzC03
s o l u t i o n and centrifuged again. The aqueous phase was
heated a t 6 0 " f o r 1 0 m i n u t e s w i t h methanol and
dimethylsulphate, cooled and M-acetate b u f f e r s o l u t i o n
was added t o a d j u s t t o pH 5.2.
The m e t h y l a t e d
sulphonylureas were e x t r a c t e d w i t h hexane and t h e
e x t r a c t was evaporated t o dryness a t 50' i n a stream o f
nitrogen. The residue was dissolved i n CS2-CHC13 (l:l),
25 u l and 2 p l was submitted t o GLC on a glass column
(61 cm X 3 mn) containing 0.5% o f PEG 20 M on Gas-Chrm
Q (80 t o 100 mesh) and the temperature was programed
f o r 190 t o 240' a t 5 min-1, w i t h helium as c a r r i e r gas
(90 m l min-1) and flame i o n i s a t i o n d e t e c t i o n . Peak
heights were compared. A t concentrations o f 10 t o 40 ug
m l - 1 i n plasma. The mean recoveries (8 determination)
were : f o r acetohexamide 9.9 and 39.4 c(g m l - 1 ; f o r the
metabolite hydroxyhexamide 14.1 and 40 c(g m l - 1 .
Fricke (32) presented a GLC method f o r t h e analysis o f
t h e drug and o t h e r drugs i n pharmaceuticals, u s i n g
s i m p l e e x t r a c t i o n s and semiautomated g a s - l i q u i d
chromatography, using Ddxil 300 as the l i q u i d phase and
an automatic sample i n j e c t o r . Results by t h i s method
and t h e o f f i c i a l and o t h e r a p p l i c a b l e methods a r e
compared. Content uniformity analysis can be made by
u s i n g t h i s procedure. The e x t r a c t i o n and chromatographic conditions were standardized t o make possible a
successful interlaboratory study.
4.3.3

Hinh-Performance L i a u i d ChrmatoqraDhy
( HPLCl

A simple HPLC assay o f t h e drug i n plasma was

developed by T a k a g i s h i e t a7 ( 3 3 ) . A sample was
extracted w i t h a mixture o f benzene and e t h y l acetate
a t pH 5 and t h e organic phase was evaporated. A 50%
s o l u t i o n i n CH3CN o f the residue was chromatographed
u s i n g a Lichrosorb RP-8 reversed-phase column and a
mobile phase composed o f 0.2% a c e t i c a c i d - methyl
c y a n i d e ( 1 : l ) . The m e t h o d c a n be u s e d f o r
b i o a v a i l a b i l i t y and c l i n i c a l pharmacokinetic studies o f
acetohexamide.

Beyer (34) used high speed l i q u i d chromatography f o r

analysis o f the drug and other a n t i d i a b e t i c agents. The
reocvery o f the drug from i n e r t t a b l e t ingredients by

ACETOHEX AM I DE

33

t h i s method was near 100%. A column (100 cm X 2 . 1 mm)

packed w i t h 1% o f ethylene-propene copolymer on Zipax
was used w i t h mobile phase o f 0.01 M disodium hydrogen
c i t r a t e containing 15% o f methanol (pH 4 . 4 ) . Detection
was c a r r i e d o u t a t 254 nm and pack a r e a s were
integrated.
Testosterone, chlorpropamide i n 95% ethanol were used
i n t e r n a l s t a n d a r d s . The p r o c e d u r e was a p p l i e d t o
compressed t a b l e t s , t h e powdered sample being e x t r a c t e d
w i t h t h e i n t e r n a l standard s o l u t i o n . Recoveries o f
added sulphonylurea were 98.9% t o 100.2%.
5.

PHARMACOKINETICS
5.1

Introduction

Acetohexamide i s used as an o r a l a n t i d i a b e t i c
agent f o r t h e t r e a t m e n t o f k e t o a c i d o s i s - r e s i s t a n t
d i a b e t e s . I t i s an i n t e r m e d i a t e a c t i n g s u l f o n y l u r e a
d e r i v a t i v e . The c l i n i c a l e f f e c t s o f lowering elevated
blood glucose l e v e l s i s s i m i l a r f o r a l l o f t h e
sulfonylurea d e r i v a t i v e s . Acetohexamide, however, i s
t h e only one t o a l s o possess u r i c o s u r i c a c t i v i t y and
t h e r e f o r e i s a p r e f e r a b l e agent t o t r e a t d i a b e t i c
p a t i e n t s w i t h gout.
The d u r a t i o n o f a c t i o n o f acetohexamide (12-24 hours)
permits once o r t w i c e d a i l y dosage. The crossover study
o f Fox
e t a7. (35) conducted i n 36 p a t i e n t s w i t h
m a t u r i t y onset diabetes m e l l i t u s i n d i c a t e d t h a t both
chlorpropamide and acetohexamide gave s i m i l a r responses
based on f a s t i n g blood sugar. Acetohexamide was used i n
a dose range o f 500-3,000 mg/day and i t i s i n d i c a t e d
t h a t primary f a i l u r e on acetohexamide i s more l i k e l y t o
respond t o chlorpropamide and v i c e versa. Appropriate
dosing r e q u i r e i n d i v i d u a l i z a t i o n o f therapy t i t r a t e d t o
t h e d e s i r e d t h e r a p e u t i c e f f e c t . The usual PO dosage
range i s 250-1500 mg/day i n s i n g l e o r d i v i d e d doses
(36,37), w i t h a maximum recommended dose o f 1500
mg/day. The 250 mg dose o f acetohexamide i s equivalent
t o 500 mg t o l b u t a m l d e , 100 mg tolazamide, o r 100 mg
chlorpropamide (36). The o r a l a n t i d i a b e t i c agents prove
more u s e f u l when d i e t a r y r e s t r i c t i o n and w e i g h t
reduction accompany t h e i r use.

ABDULLAH A. AL-EADK AND HUMEIDA A. EL-OBEID

34

Acetohexamide i s l a r g e l y metabolized t o an a c t i v e
metabolite which is excreted i n t h e u r i n e (see below).
Therefore, dosage adjustment o r t o t a l avoidance i s
necessary i n c e r t a i n cases. One such case i s t h e renal
f a i l u r e . Azotenic p a t i e n t s may experience prolonged
hypoglycemia. A t w i c e d a i l y dose i s recommended f o r
p a t i e n t s w i t h m i l d r e n a l f a i l u r e and p a t i e n t s w i t h
moderate t o severe renal f a i l u r e should not receive t h e
drug (38,39).
Dosage adjustment may a l s o be required i n p a t i e n t s with
1i v e r i n s u f f i c i e n c y since acetohexamide i s e x t e n s i v e l y
metabolized i n t h e l i v e r . Prolonged hypoglycemia may
r e s u l t i n p a t i e n t s w i t h severe l i v e r impairment (36).
Dosage r e d u c t i o n may be r e q u i r e d i n e l d e r l y o r
d e b i l i t a t e d p a t i e n t s , due t o renal o r l i v e r impairment
o r hyperresponsiveness (36).
I t i s recommended by Bennett e t a7. (39) t h a t no dosage
supplementatlon i s r e q u i r e d i n p a t i e n t s f o l l o w i n g
p e r i t o n e a l d i a1y s i s

L i k e other o r a l a n t i d i a b e t i c agents, acetohexamide may

be used i n combination w i t h i n s u l i n t o reduce i n s u l i n
r e q u i r e m e n t s i n i n s u l i n dependent m a t u r i t y o n s e t
d i a b e t i c s and t o r e d u c e t h e p o t e n t i a l f o r a
hypoglycemic reaction.
5.2

Mechanism o f Action

Acetohexamide i s a sulfonylurea d e r i v a t i v e , t h a t
produces i t s hypoglycemic e f f e c t by s t i m u l a t i n g t h e
i s l e t t i s s u e t o s y n t h e s i z e and r e l e a s e endogenous
i n s u l i n ( 4 0 ) . The h y p o g l y c e m i c e f f e c t s a r e a l s o
a t t r i b u t e d t o an increased s e n s i t i v i t y o f i n s u l i n
receptors as w e l l as improved peripheral u t i l i z a t i o n o f
i n s u l i n (37).
A r e p o r t by Lebowitz and Feinglos (41) i n d i c a t e s t h a t ,
d u r i n g chronic administration, p a r t o f t h e hypoglycemic
action o f the sulfonylureas i s e x t r a pancreatic.
Peripheral t i s s u e s may become more s e n s i t i v e t o a f i x e d
dose o f an a d m i n i s t e r e d hormone p o s s i b l y due t o an
increase i n the number o f i n s u l i n receptors.
A study on the mode o f a c t i o n o f t h e sulfonylureas (42)

has shown t h a t acetohexamide increased glucose uptake

ACETOHEXAMIDE

35

by r a t diaphragm, i n h i b i t e d the a c t i v i t y o f glucose-6phosphatase, triosephosphate isomerase and l i p o p r o t e i n

1ipase

5.3

Onset and Duration o f Action

B r e i d a h l e t a 7 . ( 4 3 , 4 4 1 r e p o r t e d a peak
hypoglycemic e f f e c t t o occur between 8 t o 10 hour post
ingestion o f acetohexamide.
A duration o f action o f 12 t o 24 hours i s reported by
Breidahl et a7. (43,44) and Galloway et a7. (45) which
i s s i m i l a r t o t h a t o f tolazamide (up t o 24 hours), less
than t h a t o f chlorpropamide (60 hour) and greater than
t h a t o f tolbutamide (6 t o 12 hours) (37).

The serum c o n c e n t r a t i o n s i n d i a b e t i c p a t i e n t s
responding w e l l t o t h e drug had mean acetohexamide
l e v e l s o f 3.7 mg/dL w i t h a ragne f o 2.5 t o 4 . 9 mg/dL
f o l l o w i n g dosage regimens o f 0.5 t o 3 g/day (46). No
good c o r r e l a t i o n between b l o o d c o n c e n t r a t i o n s o f
acetohexamide and therapeutic e f f e c t i s established.
However, f a s t i n g blood glucose concentrations a r e
decreased i n a dose-dependent f a s h i o n i n t h e dosage
range between 250 mg t o 1,000 mg (47).
5.4

AbsorDtion

O r a l l y a d m i n i s t e r e d acetohexamide i s almost
completely absorbed (47). I t i s reported t o appear i n
the blood w i t h i n 30 minutes a f t e r PO administration and
peak l e v e l s occur a f t e r 3 t o 5 hours (43,44). Galloway
et a7. (45) reported t h a t , f o l l o w i n g single PO doses o f
1 g o f acetohexamide, mean peak blood l e v e l s o f t h e
drug t o be 47 mcg/ml and f o r hydroxyhexamide mean
l e v e l s o f 60.3 mcg/ml were achieved. These peak l e v e l s
occurred w i t h i n 1.5 t o 2 hours f o r the parent compound
versus 2 t o 6 hours f o r th e a c t i v e metabolite,
hydroxyhexamide.
5.5

Distribution

J u d i s ( 4 8 , 4 9 ) r e p o r t e d t h a t acetohexamide
extensively binds t o plasma proteins t o the extent o f
65 t o 90%.

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

36

5.6

Hetabol ism

A c e t o h e x a m i d e is mainly metabolized b y
hydroxylation reactions in the liver to inactive and
active metabolites. The primary metabolite (47 to 60%)
is hydroxyhexamide (47,501. It is an active metabolite
and is reported (45,50) to be excreted unchanged in the
urine, as well as metabolized to the inactive
dihydroxyhexamide (38).
Hydroxyhexamide, like acetohexamide, possesses both
hypoglycemic and uricosuric properties (51,52), but it
is 2.5 times as potent as its parent d r u g (36).
Impairment of hydroxyhexamides el imination has been
reported (51) to result in severe hypoglycemia.
Kojima et a7. (53) investigated the effect of various
drugs on the i n v i v o metabolic reduction of
acetohexamide. Most of the nonsteroidal antiinflammatory drugs inhibited the acetohexamide
reduction in liver, kidney and heart cytosol from
rabbits. Ketone-containing drugs including warfarin
also inhibited the reduction reaction in both the liver
and the kidney; in the heart, acetohexamide reduction
was inhibited only by warfarin.
Species differences in the i n v i t r o metabolic reduction
of acetohexamide were studied (54) in rabbit, guinea
pig, hamster, rat and mouse. The rabbit exhibited the
highest acetohexamide reductase activity in the cytosol
of the liver and kidney among the species tested. The
sensitivity to specific inhibitors of cytosolic
acetohexamide reductase in the liver and kidney of the
rabbit were different from those of the rat. Only rats
and guinea pigs showed significant activity of
acetohexamide reductase activity in the microsomes of
the liver and kidney.
Nagamine e t a7 (55) estimated the rates of available
fraction for 4-acetamidoacetophenone, 4-acetylbenzenesulfonamide, and acetohexamide and their respective
reduced compounds, 4-substituted a-hydroxyethylphenyl
derivatives, in rats. The study indicated that the
compounds are i n a reversible drug-metabolite
relationship. The pharmacokinetic profiles o f the
agents were studied after an intraportal administration

ACETOHEXAMIDE

i n comparison w i t h those a f t e r I . V .
using an interconversion model.
5.7

37

administration

Excretion

Acetohexamide and i t s m e t a b o l i t e s a r e m a i n l y
e x c r e t e d b y t h e k i d n e y s . The u r i n a r y r e c o v e r y o f
radioactivity a f t e r the administration o f oral
14C-labeled acetohexamide averaged 71.6% i n 24 hours
(45). Approximatley one-half t o two-third o f t h e drug
was r e p o r t e d t o be excreted i n u r i n e as t h e a c t i v e
metabolite, hydroxyhexamide (45,501. Fecal excretion o f
r a d i o a c t i v i t y f o l l o w i n g o r a l administration o f t h e drug
i n one p a t i e n t was 15%. Even a f t e r 1 g I . V . dose
u r i n a r y recovery was o n l y 85% ( 4 5 1 , suggesting t h a t
b i l i a r y e x c r e t i o n represents a secondary r o u t e o f
e l i m i n a t i o n o f acetohexamide and/or i t s metabolites.
However, more data are needed t o confirm the occurrence
o f b l l i a r y excretion.
5.8

Half-Life

F o l l o w i n g o r a l a d m i n i s t r a t i o n o f 14C-labeled
acetohexamide t o human subjects, a mean blood h a l f - l i f e
o f t h e drug o f 1 . 6 hours was determined, using isotope
d i l u t i o n a n a l y s i s , w i t h a range o f 0 . 8 - 2 . 4 hours
(45,56).

F i e l d e t a l . ( 5 1 ) , however, reported a range o f 21-70

minutes averaging t o a value o f 55.8 minutes. The
combined h a l f - l l f e o f t h e parent compound and i t s
a c t i v e metabolite, hydroxyhexamide, i s reported t o be
5.3 hours (43-45).
The h a l f - l i f e o f acetohexamide i s
reported be prolonged i n renal f a i l u r e ( 3 8 ) .
The a c t i v e m e t a b o l i t e , hydroxyhexamide i s r e p o r t e d
(45,561 t o have a mean h a l f - l i f e o f 5.3 hours with a
range o f 3.7-6.4 hours. The average value o f 5.3 hours
agrees w i t h t h e f i n d i n g o f F i e l d e t a l . ( 5 1 ) who
reported a range o f 3.2-7.6 hours.
The blood and u r i n e data reported by Galloway et a l .
(45) agree w i t h those reported by Scheldon et a l . (46)
and confirm the report by Smith e t a l . ( 5 6 ) , t h a t the
combined half-1 i f e o f acetohexamide and hydroxyhexamide
i s comparable w i t h t h a t o f tolbutamide.

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

38

ACKNOWLEGEMENT
The authors would l i k e t o thank M r . Tanvir A. B u t t
f o r t y p i n g t h i s manuscript.
REFERENCES
1.

The Merck Index, Tenth E d i t i o n ,

Rahaway, New Jersey, 1983, page 9.

2.

The B r i t i s h PharmacoDoeia, HM S t a t i o n a r y O f f i c e ,
London, 1988, Vol. 1, page 18.

3.

B r i t . Pat. 912, 789 (1962 t o L i l l y )

No. 1.

4.

F.J.
M.A.

5.

Merck & Co.

Inc.,

Through reference

Marshall, M.V. Sigal, H.R. Sullivan, C. Cesnik and

Root, J. Med. Chem., 5 , 60 (1963).

P. Girgis-Takla and

29, 640 (1977).

I. Chroneos, J. Pharm. Pharmacol.,

6.

K. Kuroda, T. Yokoyama, T. Umeda and V.

Chem. Pharm. Bull., 26, 2565 (1978).

7.

A.

8.

B.W. Mueller and M.

77 (1979).

9.

B.W.

Burger, Sci. Pharm.,

s, 207

Takagishi,

(1978).

Lagas, Pharm. Weekbl. Sci. Ed.,

Mueller and M. Lagas,

u,1, 449

1,

(1979).

10.

T. Yokoyama, T . Umeda, K. Kuroda, K. S a t 0 and V.

Takagishi, Chem. Pharm. Bull., 27, 1476 (1979).

11.

E . G r a f , C. Beyer and 0.
Technol., 5 , 9 (1979).

12.

E. Graf, C. Beyer and 0. Abdallah, @id

28,
, 73 (1982).

13.

E. Graf, C.
(1982).

Beyer and 0. A b d a l l a h ,

ibid,

14.

E. G r a f ,
(1982).

Beyer and 0. A b d a l l a h ,

m, 28,

C.

Abdallah,

A c t a Pharm.

a,131
225

39

ACETOHEXAMIDE

Kassem, A.M. F o u l i , S. Said and E. Shehata, B u l l .

Fac. Pharm. (Cairo Univ.), 19, 309 (1982).

15.

A.A.

16.

Mfg.

17.

S u r a j P. Agarwal and Mohammed I.Wa ash;

Pharm., 3 4 ( 5 ) , 109-111 (1972).

18.

Jose Dobrecky and Rogelio J . C a l l e j a ; Rev. Fac. Quim

Farm., [Univ. Cent. Ecuador], El(16); 44 49 (1969).

19.

M.M. Amer and M . I . Walash; B u l l . Fac. Pharm..

Univ., l 2 ( 2 ) , 199-209 (1973).

Chem.,

34, 454, 467 (1963).

Cairo

W, l2(2),223-233 (1973).

Amer and M.I.

Walash:

20b. M.H. h e r and M.I.

(Pub. 1975).

Walash;

20a. M.M.

I n d i a n J.

m, l 2 ( 2 ) ,

189-198 (1973)

21.

L i l o 0. Guerello and Jose Dobrecky; Rev. Asoc. Bioauim.

Argent. 33(178-1791, 185-8 (1968).

22.

Meier, S.O. Kohor, O.F. P i e r a r t , S . S . J . Cortes;

Rev. Real. Acad. Cience. Exactas. Fis. Natur. Madrid,
6 5 ( 3 ) , 653-674 (1971).

23.

S.G.

24.

Edward F. Salem and W.W.

385-386 (1967).

25.

M a r i a K u h n e r t - B r a n d s t a e t t e r , Adel h e i d K o f l e r , A.
Vlachopoulas and A. Lobenwein; S c i . Pharm., 38(3)
154-163 (1970).

26.

D.S.

27.

Pamela Gergis-Takla and Ioannis Chroneos; Analyst,

(1235) 117-123 (1979).

104

28.

A.A.

Al-Badr
(1982).

378

29.

Pamela Gergls-Takla and Shanta Raj Joshi;

Anal. 1 ( 2 ) , 189-193 (1983).

N.G.

Solomonova and L . Z .
(Kiev), 2, 80-82 (1977).

Lazaryan,
(1980).

Dvornitskaya;

H i l t y ; J. Pharm. Sci.,

Farmatsiya (Moscow),

and S.E.

Farm.

Ibrahim;

29(2)

Pharmazie;

56(3),

36-38,

37(5),

J.

Zh.

Biomed.

ABDULLAH A. AL-BADR AND HUMEIDA A. EL-OBEID

40

30.

K.H. Surborg and E. Roeder; Pharmazie, =(7),

(1 97.3).

31.

u,
m(5), 635-638

32.

Fred L. F r i c k e , J.
1162-1 167 (1972).

33.

Yasushi Takagishi; K o j i Sato; Keizo Tomita and Teruo

Sakamoto; Yakusaku Zasshi, B ( 9 ) , 961-963 (1979).

34.

W i l l i a m F. Beyer; Anal. Chem., 44(7),

35.

O.J. Fox e t a l , J. Med. Assoc.

(1968).

36.

Product Information: Acetohexamide,

Indianapolis, 1N: 1983.

37.

AMA Department o f Drugs: AMA Drug Evaluations, 4 t h ed.

American Medical Association, Chicago, I L , 1980.

38.

B.D. Cohen, J.A. Galloway, R.E.

Med. Sci., 254, 608 (1967).

39.

W.M. Bennett, G.R. Aronoff, G.

Kidney Dis., 3, 155 (1983).

40.

A.G.

41.

H.E. Lebowitz and M.N.

(1978).

Feinglos, Diabetes Care,

42.

K.T. Augusti and P.A.

36 (1969).

Kurup, Indian J. Biochem.,

43.

H.D. Breidahl, G.C.

79 (1972).

Ennis, F . I . Martin et a l , Drugs,

44.

H.D. Breldahl, G.C.

204 (1972).

Ennis, F.I.

45.

J.A. Galloway, R.E. McMahon, H.W. Culp, F.J.

and E.C. Young , Diabetes, 16, 118 (1967).

J.W.

Kleber, J.A.

Galloway and B.E.

(1977).
Ass.

Off.

485-486,

Rodda; J.

Anal.

Pharm.

Chem.,

55(6),

1312-1314 (1972).

Alabama,

31,

1155

E l i L i l l y & Co.,

McMahon et a l , Am.

J.

Morrison et a l , Am.

J.

Gillman, L.S. Goodman and A. Gillman; Goodman and

Gillmans The Pharmacological Basis o f TheraDeutics,
MacMillan & Co. New York. 1980.

Martin et a l ,

1,

189

6(1),

3,

M, 3 ,
Marshall

ACETOHEXAMIDE

41

m, l4,

46.

J. Scheldon, J. Anderson and L. Stoner,

47.

R.E.

48.

J. Judis, J. Pharm. Sci., 6 l , 89 (1972).

49.

J. Judis,

50.

R . E . McMahon, F . J .
Pharmacol. EXD. Ther.,

51.

J.B.

52.

T.F. Yu, L. Berger and A.B.

(1968).

53.

Y. Kojima, Y. Imamura and M. Otagiri, Yakusaku Zasshi,

108(1), 66 (1988).

54.

Y. Imamura, Y. Kojima and M.

Bull., 36(1), 4199 (1988).

55.

S.

56.

(1965).

Ferner and S.

379 (1987).

w, 62,

Chaplin,

M a r s h a l l and H.W.

149, 272

(1965).

Culp,

Gutman, Metabolism,

Otagiri,

4612 (1988).

Smith, T.J.

229 (1965).

2,

J.

Boyle e t a l , N. Ensl. J. Med.,

17, 309

Chem.

Nagamine, T . Otawa, H. Nakae and S. Asada,

36(11),

D.L.

Pharmacokinet.,

232 (1973).

Field, M. Ohta, C.
277, 889 (1967).

14,

Clin.

362

Vecchio and A.A.

Pharm.

M,

Forist, Metabolism,