determination of atenolol combinations with hydrochlorothiazide and chlorthalidone in tablet...

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Determination of AtenololCombinations withHydrochlorothiazide andChlorthalidone in TabletFormulations by Reverse-PhaseHPLCS. I. Sa'sa' a , I. M. Jalal b & H. S. Khalil ba Chemistry Department, Yarmouk University, Irbid,Jordanb Al-Hikma Pharmaceuticals, P. O. Box, 182400,Amman, JordanPublished online: 19 Dec 2006.

To cite this article: S. I. Sa'sa' , I. M. Jalal & H. S. Khalil (1988) Determinationof Atenolol Combinations with Hydrochlorothiazide and Chlorthalidone in TabletFormulations by Reverse-Phase HPLC, Journal of Liquid Chromatography, 11:8,1673-1696

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JOURNAL OF LIQUID CHROMATOGRAPHY, 11(8), 1673-1696 (1988)

DETERMINATION OF ATENOLOL

THIAZIDE AND CHLORTHALIDONE IN

PHASE HPLC

COMBINATIONS WITH HYDROCHLORO-

TABLET FORMULATIONS BY REVERSE-

S. I. Sa’sa’l, I. M. Jalal2, and H. S. Khali12 ]Chemistry Department

Yarmouk University Irbid, Jordan

2A Z-Hikma Pharmaceuticals P. 0. Box 182400 Amman, Jordan

AB Si’KACT

A h i g h p e r f o r m a n c e l i q u i d c h r o m a t o g r a p h i c p r o c e d u r e i s p r e s e n t e d f o r t he s i m u l t a n e o u s d e t e r m i n a t i o n o f a t e n o l o l , h y d r o c h t a b l e t s i n me i n t e r n a

p h a s e ( 5 urn)

o r o t h i a z i de, a n d c h l o r t h a l i done i n p h a r m a c e u t i c a I c o m b i n a t i o n s . An a l i q u o t o f t h e s a m p l e i s d i s s o l v e d

h a n o l c o n t a i n i n g m e t h y l p - h y d r o x y b e n z o a t e as an s t a n d a r d and c h r o m a t o g r a p h e d on a s v p e l c o s i l

LC-8-DB (250mn x 4.6mn i .d . ) c o l u m n u s i n g a m o b i l e f 1.0 amnonium a c e t a t e and 2.0 nfti o c t a n e s u l f o n i c

a c i d s o d i u m s a l t i n a c e t o n i t r i l e : w a t e r ( 2 5 : 75) s o l u t i o n , t h e pH was a d j u s t e d t o 3.5 w i t h g l a c i a l a c e t i c a c i d . The r e l a t i v e s t a n d a r d d e v i a t i o n s a r e l e s s t h a n i ?/J f o r t h e f i v e c o m n e r c i a l t a b l e t s a n a l y z e d . The m e t h o d was t e s t e d f o r l i n e a r i t y , r e c o v e r y , and s p e c i f i c i t y and was f o u n d t o b e f a s t , s t a b i l i t y - i n d i c a t i n g , a n d f r e e f r o m i n t e r f e r e n c e s .

1673

Copyright 0 1988 by Marcel Dekker, Inc.

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1674

I NTRODUCT I OM

SA’SA’, JALAL, AND KHALIL

Combina t ion t a b l e t fo rms o f a t e n o l o

h y d r o c h l o r o t h i a z i d e ( I I ) , and a t e n o l o l w i t h

( I l l ) a r e m a r k e t e d by s e v e r a l m a n u f a c t u r e r s as

a n t i h y p e r t e n s i v e drugs .

( 1 ) w i t h

h l o r t h a l i d o n e

d i u r e t i c and

A t e n o l o l i s a b e t a - a d r e n e r g i c r e c e p t o r a n t a g o n i s t .

A n a l y s i s o f a t e n o l o l has p r e v i o u s l y been d e s c r i b e d by

s p e c t r o f l u o r o m e t r i c me thod(1 ) . The method i s s i m p l e b u t c o u l d

n o t be a p p l i e d t o i t s c o m b i n a t i o n s w i t h h y d r o c h l o r o t h i a z i d e

and c h l o r t h a l i d o n e . The GLC d e t e r m i n a t i o n o f I i s s p e c i f i c

and s e n s i t i v e b u t r e q u i r e s a l e n g t h y d e r i v a t i z a t i o n s t e p (2).

t r e a

ava i

w i t h

H y d r o c h l o r o t h i a z i d e and c h l o r t h a l i d o n e a r e w i d e l y used

as d i u r e t i c s i n t h e t r e a t m e n t o f h y p e r t e n s i o n and i n t h e

ment o f c o n g e s t i v e h e a r t f a i l u r e . S e v e r a l methods a r e

a b l e f o r t h e assay o f I I a l o n e (3 -51 , o r i n c o m b i n a t i o n

o t h e r d rugs such as m e t h y l d o p a ( 6 ) , h y d r a l a z i n e ( 7 ) ,

r e s e r p i n e ( 7 , 8 ) , f u r o s e m i d e ( 9 ) , t r i m a t e r e n e ( l 0 1 , and o t h e r -

o f t h e t h i a z i d e f a m i l y ( 1 1 ) . However,

i s a p p l i c a b l e t o t h e s imu l taneous

compounds i n c o m b i n a t i o n w i t h 1 .

a n t i h y p e r t e n s i v e d rugs

none o f t h e methods

de t e r m i na t i on o f t hese

T h i s paper desc r

q u a n t i t a t i v e d e t e r m i n a

bes a r e v e r s e phase HPLC assay f o r t h e

i o n o f I, I I and I l l and some o f t h e i r

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DETERMINATION OF ATENOLOL COMBINATIONS 1675

OH I

OCH&HCH,NHCHMe2

H CHZC NH, @ II 0

I I1

0 v

COOCH, I

OH Internal Standard

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1676 SA’SA’, JALAL, AND KHALIL

r a l a t e d compounds 14-amino-6-chloro-l,3-benzenedisulfonamide

( I V ) and 2-(4-chloro-3-sulphamoylbenzoyl~ b e n z o i c a c i d ( V ) ! .

The assay was a p p l i e d s u c c e s s f u l l y t o f i v e comnerc ia l

p r o d u c t s and p r o v e d t o be f r e e o f i n t e r f e r e n c e s f r o m

e x c i p i e n t s n o r m a l l y used i n t a b l e t f o r m u l a t i o n s . The assay i s

f a s t s i n c e i t r e q u i r e s o n l y l i t t l e sample m a n i p u l a t i o n .

I t was a l s o v a l i d a t e d f o r p r e c i s i o n and accuracy . The

pa ramete rs i n v e s t i g a t e d i n t h i s assay i n c l u d e d a c e t o n i t r i l e

pe rcen tage , pH, and i o n i c s t r e n g t h o f t h e m o b i l e phase.

EXPERIMENTAL ------------

Appa r a t u s ---------

The a p p a r a t u s employed was a V a r i a n 2010 pump ( V a r i a n

A s s o c i a t e s , Inc . P a l o A l t o , CA, U.S.A.! equ ipped w i t h a 10-uL

l o o p i n j e c t o r model 7125 (Rheodyne, C o t a t i , Ca USA)

connected t o a V a r i a n 2050 s p e c t r o p h o t o m e t r i c d e t e c t 0 and a

V a r i a n 4290 i n t e g r a t o r . A r e v e r s e phase column(250mn x 4 . 6 m

i.d.) S u p e l c o s i l LC-8-DB (5 urn) ( s u p e l c o , Inc., Pennsy lvan ia ,

USA) was used a t ambien t t empera tu re .

Ma te r i a I s --------- The r e f e r e n c e s tandards , and t h e i n t e r n a l s t a n d a r d

( m e t h y l p -hydroxybenzoate , were f r o m BP ( B r i t i s h

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Pharmacopoeia Comni s s i o n L a b o r a t o r y , M i d d l e s e x , U.K.).

A c e t o n i t r i le-HPLC, m e t h a n o l , and g l a c i a l a c e t i c a c i d were

f r o m May & Baker L t d (Dagenham, U.K.), R iede l -de Haen AG

(West Germany) and Koch-L igh t L t d ( H a v e r h i l l , U.K.),

r e s p e c t i v e l y . h o n

a c i d sod ium s a l t

( S w i t z e r l a n d ) . Wate

um a c e t a t e purum g rade and o c t a n e s u l f o n i c

(99.0 %) were o b t a i n e d f r o m F l u k a

was a lways d i s t i l l e d and d e i o n i z e d .

E x c i p i e n t s u s u a l l y used i n the t a b l e t f o r m u l a t i o n s :

magnesium ca rbona te , p o t a t o s t a r c h , sod ium l a u r y l s u l f a t e ,

pov idone m i c r o c r y s t a l l i n e c e l l u l o s e , magnesium s t e a r a t e , and

c o l o r i n g agen ts were s u p p l i e d b y Al-Hikma P h a r m a c e u t i c a l s ,

h a n - J o r d a n . Commercial t a b l e t s were purchased l o c a l l y .

P r e p a r a t i o n o f Re fe rence D e c o m p o s i t i o n P r o d u c t s : ................................................

The d e c o m p o s i t i o n p r o d u c t s o f h y d r o c h l o r o t h i a z i d e and

c h l o r t h a l i d o n e ( I V and V) were p r e p a r e d (Scheme I) by

d i s s o l v i n g 1.0 gm o f t h e p a r e n t compound i n 30 m l o f 20 %

NaOH s o l u t i o n . The m i x t u r e s w e r e ' r e f l u x e d f o r 2 hrs, a l l o w e d

t o s t a n d f o r 20 h r s , and t h e n a c i d i f i e d t o pH 4.0 w i t h 6 N

HCI. The r e s u l t i n g p r e c i p i t a t e s were s e p a r a t e d b y vacuum

f i l t r a t i o n , washed w i t h d i s t i l l e d w a t e r , and r e c r y s t a l l i z e d

f r o m w a t e r f o r IV and m e t h a n o l : w a t e r f o r V t o a c o n s t a n t

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1678 SA'SA', JALAL, AND KHALIL

Scheme 1

melting point. T h e identity of the products (IV and V) w a s

conf i rmed by I R and NMR.

Chromatographic Conditions: ........................... T h e m o b i l e phase consists of 1.0 nM amnonium acetate and

2.0 &I sodium octanesulfonate n acetonitri l e : water ( 2 5 : 7 5 ) ;

the pH w a s adjusted to 3.5 w th glacial acetic acid. T h e

m o b i l e phase was always filtered using 0.45 um-membrane

filters (Supelco, Inc.), and degassed by vacuum prior to use.

T h e flow rate was 1.5 mL/min. T h e wavelength w a s 254 nm and

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t h e s e n s i t i v i t y was s e t a t 0.2 AUFS. The c h a r t speed was 0.25

cm/mi n.

S tudy I n t e r f e r e n c e s P lacebo E x c i p i e n t s - A

m i x t u r e o f t h e e x c i p i e n t s was d i s s o l v e d and t r e a t e d i n t h e

same manner as t h e sample s o l u t i o n . Ten-ul i n j e c t i o n s were

made under t h e ch romatog raph ic c o n d i t i o n s d e s c r i b e d .

I n t e r n a l S t a n d a r d S o l u t i o n - The i n t e r n a l s t a n d a r d

s o l u t i o n was p r e p a r e d b y d i s s o l v i n g 1Omg o f m e t h y l p-hydroxy-

benzoa te i n 1 .O L o f me thano l .

S tandard S o l u t i o n s For L i n e a r i t y - S t a n d a r d s t o c k

s o l u t i o n c o n t a i n i n g 150.0 mg o f A t e n o l o l , 37.5 mg o f

h y d r o c h l o r o t h i a z i d e , and 37.5 mg o f c h l o r t h a l i d o n e was

p r e p a r e d i n 25 m l o f the i n t e r n a l s t a n d a r d s o l u t i o n . The

f o l l o w i n g c o n c e n t r a t i o n s o f a t e n o l o l , h y d r o c h l o r o t h i a z i d e ,

and c h l o r t h a l i d o n e i n t h e i n t e r n a l s t a n d a r d s o l u t i o n were

p r e p a r e d by d i l u t i o n : 5.00, 7.00, 8.00, 10.00 and 15.00,

1.25, 1.50, 2 -00 , 2.50, and 3.75 ; and 1.25, 1.50, 2.00,

2.50, and 3.75 ug / lOuL , r e s p e c t i v e l y .

A t e n o l o l and C h l o r t h a l i d o n e S t a n d a r d S o l u t i o n - One

hundred mg o f a t e n o l o l and 25.0 mg o f c h l o r t h a l i d o n e were

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d i s s o l v e d i n 25.0 rnL i n t e r n a l s t a n d a r d s o l u t i o n . T h i s was

f u r t h e r d i l u t e d w i t h t h e i n t e r n a l s t a n d a r d s o l u t i o n t o o b t a i n

a f i n a l c o n c e n t r a t i o n o f 1.0 mg/mL and 0 .25 mg/mL,

r e s p e c t i v e l y .

H y d r o c h l o r o t h i a z i d e S t a n d a r d S o l u t i o n - Twenty f i v e mg

h y d r o c h l o r o t h i a z i d e was d i s s o l v e d i n 25 mL i n t e r n a l s t a n d a r d

s o l u t i o n . T h i s was f u r t h e r d i l u t e d w i t h t h e i n t e r n a l s t a n d a r d

s o l u t i o n t o o b t a i n a f i n a l c o n c e n t r a t i o n o f 0 . 2 5 mg/mL.

C h l o r t h a l i d o n e S t a n d a r d S o l u t i o n - Twenty f i v e mg o f

c h l o r t h a l i d o n e was d i s s o l v e d i n 2 5 mL i n t e r n a l s t a n d a r d

s o l u t i on

so I u t i on

P r epa ra t

T h i s was f u r t h e r d i l u t e d w i t h t h e i n t e r n a l s t a n d a r d

t o o b t a i n a f i n a l c o n c e n t r a t i o n o f 0.25 mg/mL.

on o f The Sample S o l u t i o n : ................................... Twenty t a b l e t s (one t a b l e t i f c o n t e n t u n i f o r m i i v was t o

be d e t e r m i n e d ) were we ighed and powdered. A c c u r a t e l y we ighed

p o r t i o n s o f t h e powder (each e q u i v a l e n t t o t h e w e i g h t o f one

t a b l e t ) were p l a c e d i n 2 5 rnL-vo lumet r ic f l a s k . Each sample

was s o n i c a t e d f o r t h r e e m i n u t e s w i t h 20 r L o f t h e i n t e r n a l

s t a n d a r d s o l u t i o n , t h e n comp le ted t o vo’ume w

s t a n d a r d s o l u t i o n . Samples werz f u r the r d i

i n t e r n a l s t a n d a r d s o l u t i o n t o o b t a i n a concen

th t h e i n t e r n a l

u t e d w i t h t h e

r a t i o n o f 1 .OO,

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0.25, and 0.25 mg/mL o f a i e n o l o l , h y d r o c h l o r o t h i a z i d e , and

c h l o r t h a l i d o n e . The s c ; l u t i o n s were f i l t e r e d t h r o u g h 0.45 um-

membrane f i l t e r s .

Pe rcen t Recovery S tudy - The s t u d y was p e r f o r m e d b y

p r e p a r i n g s y t h e t i c m i x u t r e s i d e n t i c a l t o t h e p h a r m a c e u t i c a l

f o r m u l d t i o n s and were s p i k e d w i t h known amounts o f

50.0, 70.0, 80.0, 100.0, 125.0, and 150.0 rng

f o l l o w i n g amounts o f h y d r o c h l o r o t h i a z i d e and c h l o r o

a teno I o I

and t h e

h a l i d o n e :

12.50, 15.00, 20.00, 25.00, 30.00, and 37.5Omg spann ing t h e

range o f SO-lSOo/o o f t h e expec ted assay va lues . The r e s u l t i n g

m i x t u r e s were assayed and t h e r e s u l t s o b t a i n e d were compared

w i t h t h e expec ted one.

Assay Method - Equa l volumes (10-uL) and a p p r o x i m a t e l y

equa l c o n c e n t r a t i o n s o f t h e s t a n d a r d and sample s o l u t i o n s

were i n j e c t e d i n t o t h e HPLC and chromatographed under t h e

c o n d i t i o n s d e s c d r i b e d above. The s t a n d a r d and t h e sample

s o l u t i o n s c o n t a i n e d t h e same c o n c e n t r a t i o n o f t h e i n t e r n a l

s tandard . The q u a n t i t y o f each component

w i t h i n t h e l i n e a r i t y range

C a l c u i a t i o E - The r e s u l t s were ca

response r a t i o s (RR) r e l a t i v e t o i n t e r n a l

peak a reas :

RRx

n j e c t e d was a lways

c u l a t e d u s i n g

s t a n d a r d based on

P e r c e n t o f t h e l a b e l c l a i m found = ---- X 100 RR s

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Where RRx = sample response r a t i o ; RRs = s t a n d a r d response

r a t i o .

RESULTS AND.DISCUSSION ......................

I n o r d e r t o o p t i m i z e t h e c h r o m a t o g r a p h i c pa ramete rs , t h e

e f f e c t s o f a c e t o i t r i l e c o m p o s i t i o n , pH, and i o n i c s t r e n g t h on

t h e c a p a c i t y f a c t o r (k') were conduc ted ( F i g . 1 -3) . The

I l l , V, and t h e i n t e r n a l

ed b y t h e v a r i a t i o n o f

l e phase ( F i g . 1 ) . A t l ow

c a p a c i t y f a c t o r ( k l ) v a l u e s

s t a n d a r d were s u b s t a n t i a l

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

f o r I,

y a f f e c

t h e mob

a c e t o n i t r i l e c o n c e n t r a t i o n , kl v a l u e s were h i g h

peaks were broadened. A t h i g h a c e t o n i t r i l e c o n c e n t r a t

v a l u e s were lower w i t h s h a r p e r peaks. V a l u e s o f k l

and I V were s l i g h t l y a f f e c t e d b y a c e t o n i t r i l e compos

nd t h e

ons kl

f o r I 1

t i o n i n

the m o b i l e phase w i t h i n t e r f e r e n c e a t h i g h e r t h a n 30 %.

T h e r e r f o r e , a m o b i l e phase o f 25 96 a c e t o n i t r i l e was s e l e c t e d

s i n c e i t p r o v i d e d c o m p l e t e l y r e s o l v e d sha rp peaks w i t h i n a

reasonab le e l u t i o n t ime.

V a r i a t i o n o f pH ( F i g . 2 ) y i e l d e d c o n s i d e r a b l e change i n

k l v a l u e s o f V w h i l e i t had no e f f e c t on k l v a l u e s o f t h e

o t h e r compounds. compound V, b e i n g the o n l y c a r b o x y l i c a c i d ,

e x p l a i n s t h e c o n s i d e r a b l e i n c r e a s e i n k l upon i n c r e a s i n g

t h e pH va lue . T h i s b e h a v i o u r c o u l d be a t t r i b u t e d t o t h e

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16

12

10

k' 8

6

4

2

l& 2& 25% 36% 35%

CONCENTRATION OF ACETONITRILE (v/v)

F i g u r e ( 1 ) : P l o t s o f t h e c a p a c i t y f a c t o r versus t h e a c e t o n i t r i l e compostion i n t h e mobi l e phase.

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12

10

8

k’ 6

4

2

Internal Standard * 0 - -

I11 I 0

3.0 3.5 4.0 4.5 5.5

PH

F i g u r e (7) : P l o t s o f t h e c a p a c i t y f a c t o r ve rsus pH.

i n c r e a s e d s o l u b i l i t y o f a c i d a t h i g h e r pH va lues . pH 3 .5 - 4.5 c o u l d be chosen as opt imum v a l u e s f o r t h e a n a l y s i s .

However, pH 3.5 was s e l e c t e d a t w h i c h t h e i n t e r n a l s t a n d a r d

e l u t e s b e f o r e compound V. There was, however, no v a r i a t i o n i n

t h e sharpness o f t h e peaks between pH 3.5 - 4.5 and no

c o n s i d e r a b l e d i f f e r e n c e i n t h e e l u t i o n t ime.

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DETERMINATION OF ATENOLOL COMBINATIONS

8 -

6 - k'

1685

( F - 1 Internal Standard - - - -

0.5 1.0 2.0

I&! OF &?&%IUh! ACETATE

F i g u r e ( 3 ) : P l o t s o f t h e c a p a c i t y f a c t o r ve rsus t h e m o l a r i t y o f amnonium a c e t a t e .

The v a r i a t i o n o f t h e i o n i c s t r e n g t h s l i g h t l y a f f e c t e d

the k l v a l u e s ( F i g . 3 ) . S a t i s f a c t o r y k f v a l u e s were

o b t a i n e d a t 1.0 rdvl amnonium a c e t a t e . A l t h o u g h t h e e f f e c t o f

amnonium a c e t a t e on k l v a l u e s i s i n s i g n i f i c a n t , y e t i t has

a c o n s i d e r a b l e e f f e c t on r e d u c i n g t h e peak t a i l i n g o f

a t e n o l o l . A l s o , t h e a d d i t i o n o f 2.0 mM o f sod ium

o c t a n e s u l f o n a t e has been f o u n d t o reduce t h e peak t a i l i n g o f

a t e n o l o l .

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1686 SA'SA, JALAL, AND KHALIL

To d e t e r m i n e t h e l i n e a r i t y o f t h e d e t e c t o r response,

c a l i b r a t i o n s t a n d a r d s o l u t i o n s o f I, I I and I l l were p r e p a r e d

as d e s c r i b e d i n t h e t e x t . A p l o t o f peak a r e a r a t i o s ve rsus

amounts i n j e c t e d was l i n e a up t o 15.00, 3.75, and 3.75 ug,

r e s p e c t i v e l y w i t h c o r r e l a t on c o e f f i c i e n t s o f 0.999 o r b e t t e r

f o r each compound.

To d e t e r m i n e t h e accu racy o f

was s p i k e d w i t h a p l a c e b o and sub

a l i cases , s a t i s f a c t o r y r e c o v e r

t h e method, each s t a n d a r d

e c t e d t o HPLC a n a l y s i s . I n

e s and r e p r o d u c i b i I i t y o f

peak a reas were o b t a i n e d . A l i n e a r r e g r e s s i o n a n a l y s i s o f t h e

d a t a shows e x c e l l e n t c o r r e l a t i o n ove r t h e a n a l y s i s range

s t u d i e d ( T a b l e I ) . No i n t e r f e r e n c e s due t o e x c i p i e n t s were

produced. The d e t e c t i o n l i m i t s

, 1 1 , and I l l, r e s p e c t i v e l y , as

andard s o l u t i o n s w i t h me thano l

and i n j e c t i n g 10-uL o f each s o l u t i o n i n t o the column.

d e t e c t e d i n t h e chromatograms

were 200, PS, and 50 pg f o r

d e t e r m i n e d b y d i l u t i n g t h e s

The s p e c i f i c i t y o f t h e method i s i l l u s t r a t e d i n ( F i g . 4 )

where comp le te s e p a r a t i o n was n o t i c e d f o r a s y n t h e t i c m i x t u r e

o f I , I I , 1 1 1 , IV , V, and the i n t e r n a l s tandard . The r e s u l t s

o f a n a l y s i s o f f i v e c o m n e r c i a l p r o d u c t s ( T a b l e I I , F i g u r e s 5-

8 ) i n d i c a t e t h a t t h e p roposed assay can be used f o r t h e

q u a n t i t a t i o n o f I , I I , . a n d I l l i n c o m n e r c i a l samples. The

accu racy o f t h e me hod was s u p p o r t e d b y t h e c loseness o f t h e

r e s u l t s t o t h e l a b e c l a i m . The p r e c i s i o n o f t h e HPLC method

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I t+m

-

I I I

+ED

-

Adde

dlTa

blet

Fo

und/

Tabl

et

Reco

very

Adde

dITa

blet

Fo

undl

iabl

et

Reco

very

Ad

dedI

Tabl

et

FoundITablet

Reco

very

(n

s)

(mg)

(%

) (w

) (ng)

(9q

(mp)

(n

s)

!%I

50.00

M.O

l+l.7

6 -

100.

02+1

.76

12.5

0 12

.772

1.06

10

2.1

551.

06

12.5

0 12

.47M

.37

- 99

.7e.

37

70 .00

69.8

19-3

4 99.735-34

15.0

0 15

.1ot

O.7

3 -

100.

67+0

.73

15.0

0 15

.32+

0.27

-

102.

1 3M

.27

- 80

.00

80.7

2tp.

82

10

0.~

*8

2 20.00

20.1

8+1*

81

100.

9otl

.81

- 20

.00

20.0

7+0.

60

- 10

0.35

+0.6

0

100.

00

100.

2OH

l.55

- 1

00

~~

~5

5

25.0

0 25

.01M

.62

- 10

0.04

+0.6

2 25

.OO

24

.6.2

0

98.4

q10.

20

125 .00

12

4.e

.29

99

.57+

0.29

30.00

29.6

8+0.

70

- 98

-933

l.70

30 .oo

29

.96+

0.32

-

99.8

7+0.

32

- 37

.819

.43

1 OO

.83M

.43

1500

.00

148.

86to

.27

- 99

.24s

.27

37.5

0 37

.31M

.96

- 99

.499

.96

37.5

0 -

Intercept :

1.01

82

0.48

81

- 0.0

079

Slope

: 0.

9877

0.

9794

1.

0020

R : 0.9999

0,9999

0.9996

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11

F i g u r e ( 4 ) : A t y p i c a l chromatogram o f a 10-uL i n j e c t i o n o f a s y n t h e t i c m i x t u r e o f I - V and t h e i n t e r n a l s t a n d a r d .

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f ATENXCL

a Hypoz i de

b Tenoret i c

C

Esidrex

b Hygrotow-50

e Hygro tow1 00

a

100.3 + 0.04 -

100.41 2 0.55

Lot 634 (AlHikna Phamceut

Lot 1 H 323 (Imperial Chenical

Lot 006000 (CIB4 - El@' Limi

b

C

d

e

f

Lot 011m (CIW - CEIGY!

Lot 745587 (CIR4 - am)

100.82 f 0.03 -

99.27 i 0.81 -

100.06 f 0.59 -

99.83 - + 0.46

99.87 + 0.99 - -

cals - Aman - Jordan).

Industries PLC, U.K.)

ed, Basle, Ssvitzerland)

hkan 2 RSD for 3 x 6 deteminations (three sarples, six injections each).

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a -

H !

1

XI

i F i g u r e ( 5 ) : a ) A t y p i c a l chromatogram o f a 10-uL i n j e c t i o n

o f a s t a n d a r d c o n t a i n i n g 2.5 u g o f I I , 18 ug o f I , and 0.1 ug o f t h e i n t e r a l s t a n d a r d ( t h e assay method p r o c e d u r e ] .

b ) A t y p i c a l chromatogrtam o f a 10-uL i n j e c t i o n o f a comnercia l p r o d u c t ( H y p o z i d e ) .

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F i gu

- L

TIME (MIN.)

( 6 ) : a ) A t y p i c a l chromatogram o f a 10-u i n j e c t i o n o f a s t a n d a r d c o n t a i n i n g 10 u g o f I , 2.5 ug o f I I I , a n d 0.125ug o f t h e i n t e r n a l s t a n d a r d .

b) A chromatogram f o r a 10-uL i n j e c t i o n o f a s o l u t i o n made o f one t a b l e t o f T e n o r e t i c ( t h e assay method p r o c e d u r e ) .

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1 F i g u r e ( 7 ) : a ) A t y p i c a l chromatogram o f a 10-uL i n j e c t i o n

o f a s t a n d a r d c o n t a i n i n g 2.5 ug o f II and 0.1 ug o f t h e i n t e r n a l s t a n d a r d .

b ! A chromatogram f o r a 10-uL i n j e c t i o n o f a s o l u t i o n made o f one t a b l e t o f E s i d r e x ( t h e assay method p r o c e d u r e ) .

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1693 DETERMINATION OF ATENOLOL COMBINATIONS

HYGRYKN-50 HYGI#Tr(3N-100

SAMPLE m m m

8 ;r)

-c, t rn

k a, c, c H

P.1 h

r%-

b -

1

Q

C - SAMPLE -

c;

F i g u r e ( 8 ) : A t y p i c a l chromatogram o f a 10-uL i n j e c t i o n s f o r :

a ) a s t a n d a r d c o n t i n i n g 2.0 ug o f I l l and 0.1 ug o f t h e i n t e r n a l s t a n d a r d .

b ) sample o f Hygroton-50.

c ) a s t a n d a r d c o n t a i n i n g 2.5 ug o f 1 1 1 and 0.1 ug o f t h e i n t e r n a l s t a n d a r d .

d ) sample o f Hygroton-100.

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TABU

E (1

11)

TABL

ET No.

1 2 3 4 5 6 7 8 9 10

hlea

n m

H

i gh

LOW

a"r u

\IIFG

%lT

Y K

R I

, I I

, P

N)

1 I I

IN O

JvM

Rcl

PL TAELETS B

y W

LC.

PE

RC

EN

T

LA

BE

L

CL

AI

M

FO

UN

D(*

)

96 e4

2 10

4.62

99

.58

106.

56

98 -0

5 10

0.49

10

4.04

98

-66

94.56

103.

03

100.

30

3.44

104.6

5 97

.86

97.8

6 10

4.28

97

.89

103.

75

99.85

99

.21

100 -

63

99.8

4 10

0.59

10

0.84

100.

47

2.13

104.

28

94.5

6

98.9

8 10

0.00

10

3.48

99.86

100.

67

97.97

10

0.12

96.50

96 -7

6 10

2.12

99.6

5 2.2

1 10

3.48

96.50

96.32

97

.10

100.

32

97.4

9 98

.1 2

97.55

98

.31

96.32

96

.68

100.8

3

97.9

0 1.6

0 10

0 .83

96

.32

(*)

Each

Dat

a p

oin

t is

the

aver

age

of

tw

inje

ctio

ns)

.

100.

78

105 2

5

91.6

3 97

.14

98.1

1 10

0.12

94.58

10

5.50

97

.07

99.37

4.5

8 10

5.50

91

.63

103.

50

95.21

10

2.46

10

1.79

96

.53

94.4

8 10

0.22

97

.76

101.

47

97.45

97

.81

98.51

2.8

6 10

2.46

94

.a

100.

16

95.11

10

2.91

97.0

5 10

3.01

10

0.86

91

1.50

95.11

94

.89

95.1

1

98.2

7 3.3

4 10

3.01

94

.89

m z !b U

7c

X

9 c r

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i s s u p p o r t e d b y t h e v e r y s m a l l r e l a t i v e s t a n d a r d d e v i a t i o n

(RSD) based on 3 x 6 r e a d i n g s .

The s p e c i f i c i t y o f t h e method i s f u r h t e r c o n f i r m e d b y

t h e r e s u t s o f c o n t e n t u n i f o r m i t y w h i c h show comp l iance t o

s p e c i f i c a i o n s o f a l l dosage forms and s u p p o r t t h e

s p e c i f i c i t y o f t h e HPLC r e s u l t s ( T a b l e I l l ) .

and I I I by

one mounth

t h i s p e r 0

Compound I

I n

shown

a v a i l a b

p r e c i s e

p l a c i n g sa

t h e r e s u

d w i t h o u t

I degraded

A s t a b i l i t y s t u d y was p e r f o r m e d on s tandards o f I, I I ,

p l e s i n w a t e r - g l y c e r i n b a t h a t 6OoC f o r

t s show t h a t , compound I I was s t a b l e i n

any measurab le d e g r a d a t i o n t o IV .

t o g i v e t r a c e s o f V(0.06+5.33% mg).

c o n c l u s i o n , t h e HPLC assay d e s c r i b e d h e r e has been

c o m n e r c i a l l y t o be o f g e n e r a l a p p l i c a b i l i t y t o

e p r o d u c t s . The method i s q u i t e s imp e , a c c u r a t e ,

r a p i d and easy t o p e r f o r m .

ACKNClWLEDGEMENTS

The f i n a n c i a l s u p p o r t p r o v i d e d b y Yarmouk U n i v e r s i t y ,

M i n i s t r y o f P lann ing -Jo rdan , and Al-Hikma P h a r m a c e u t i c a l s ,

h a n - J o r d a n a r e g r a t e f u l l y acknowledged. The a u t h o r s wou ld

a l s o l i k e t o thank Al-Hikma P h a r m a c e u t i c a l s f o r p r o v i d i n g t h e

raw m a t e r i a l s and s tandards .

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1696

REFERENCES

SA'SA, JALAL, AND KHALIL

1 . Kaye C.M., Br. J . C l i n . Pharmacol., 1, 84 (1974) .

2. Ma1 b i c a 1 .O.and Monson K.R., J .Pharm. S c i . ,*,1992( 1975).

3 . Cohen A.I., Kee le r . B.T., Coy N.H., and Y a l e H.L., Ana l . Chem., 34, 216 (1962) .

4. D e l e o A.B. and S t e r n M.J., J.Pharm. S c i . , s , 173 (1966) .

5. F a z z a r i F.R. , J . Assoc. O f f . Anal . Chem.,E, 582 (1970) .

6. Chu R., i b i d . , 54, 603 (1971) .

7. U rbany i T. and O 'Conne l l A., Anal.Chem.,44, - 565 (1972) .

8. B u t t e r f i e d A.G.,Lovering E.G.,and Sears R.W., J . Pharm. Sci . , - 67, 650 (1978) .

9. Abd ine H., E l Sayed M.A.H., and E l Sayed Y.M., J.Assoc. O f f . Ana l . Chem., 6l, 6975 (1978) .

10. Menon G.N. and W h i t e L.B., J.Pharm. S c i . , ~ , 1 0 8 3 ( 1 9 8 1 ) .

11. K k o l o s E. and Walker J.,Anal. Chem. Acta,80,117 - (1975) .

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determination of atenolol combinations with hydrochlorothiazide and chlorthalidone in tablet...

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