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High-Performance LiquidChromatography and UV-Derivative SpectrophotometryDetermination ofAcetylsalicylic Acid andChlormezanoneD. Ivanovid a , C. Herrenkhecht b , E. Guernet-Nivaud b & M. Guernet ba Faculty of Pharmacy , Vojvode Stepe 450, 11000,Belgrade, Yugoslaviab Faculty of Pharmacy , University Paris-Sud , J. B.Clement, 92290, Chatenay-Malabry, FrancePublished online: 23 Sep 2006.

To cite this article: D. Ivanovid , C. Herrenkhecht , E. Guernet-Nivaud & M.Guernet (1992) High-Performance Liquid Chromatography and UV-DerivativeSpectrophotometry Determination of Acetylsalicylic Acid and Chlormezanone,Analytical Letters, 25:9, 1693-1703, DOI: 10.1080/00032719208018244

To link to this article: http://dx.doi.org/10.1080/00032719208018244

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ANALYTICAL LETERS, 25(9), 1693-1703 (1992)

HI GH-PERFORMANCE LIQUID CHROMATOGRAPHY AND W-DERI VAT1 VE

SPECTROPHOTOMETRIC DETERMINATION OF ACETYLSALICYLIC ACID

AND C H L 0 W K a " E

K e m r d s : High-performance liquid chromatography (HPLC), Derivative spectrophotometry, Acetylsalicylic acid, C hlo rme zan on e

D. Ivanovit?, C. Herrenkhecht'. E. G u e r n e t - N i v a u d ' and M. G u e r n e t '

'Facu 1 ty of Pharmacy , 'Faculty of Pharmacy, University Paris-Sud, Vojvode Stepe 450, 11000 Belgrade,Yugoslavia

J.B.Clement, 92290 Chatenay-Kalabry, France

ABnRACT

HPLC and UV-derivative spectrophotometry methods were

used for assaying acetylsalicylic acid and chlormezanone

in pure state and in pharmaceutical preparations as

Trancogesic@ tablets.

In this paper the results are presented using the zero-

crossing technique. In this way, the interference of two

components mixed together has been eliminated.

* To whom correspondence should be addressed

1693

CoPynght @ 1992 by Marcel Dekker. Inc.

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1694 IVANOVIC ET AL.

INTRODUCTION

Chlormezanone, 2-(4-chlorphenyl)-3-methyperhydro-l,3-

thiazin-4-one 1 , l -d ioxyde , is a t r a n q u i l i s e r used i n t h e

t r e a t m e n t of a n x i e t y and t e n s i o n s t a t e s . A number of

a n a l y t i c a l methods f o r t h e d e t e r m i n a t i o n of a c e t y l -

s a l i c y l i c a c i d and chloraezanone are r e p o r t e d .

V i s i b l e and UV-spectr ophotouietr ic*-', and chromato-

g r a p h i c methods such as thin- layerSed, gas7*' and high-

performance l i q u i d chromatography*" have been d e s c r i b e d .

We propose and compare two methods f o r t h e s imul taneous

de te rmina t ion of a c e t y l s a l i c y l i c acid and chlormezanone i n

mixture . The results obta ined w i t h RP high-performance l i q u i d

chromatography and " z e r o c r o s s i n g " d e r i v a t i v e s p e c t r o -

photometry a r e p r e s e n t e d . These methods are a p p l i e d t o

de te rmine t h e aforementioned s u b s t a n c e s i n pharmaceut ica l

formula t ions .

EXPERIMENTAL

Reagents

The s t o c k s o l u t i o n of a c e t y l s a l i c y l i c a c i d was used as a

s t a n d a r d f o r i ts d e t e r m i n a t i o n i n pharmaceut ica l

f o r m u l a t i o n s i n t h e French Pharmacopoeia. The s t a n d a r d

s u b s t a n c e of chlormezanone is a product of l a b o r a t o i r e s

Ster l ing-Winthrop, France. A l l s o l v e n t s ( acet ic acid,

methanol, "Normapur" Pro labo , France ) were of a n a l y t i c a l

g r a d e . The s o l v e n t s were deoxygenated and f i l t e r e d through a

m i l l i p o r e DA 0 . 6 5 ~ 1 1 f i l t e r . Double d e s t i l l e d water was used .

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DETERMINATION OF ACETYLSALICYLIC ACID 1695

Apparatus

The chromatographic system cons i s t ed of a Shimadzu LC-BA

pump, a Rheodyne i n j e c t o r , a Shimadzu SPD-6A d e t e c t o r

and Shimadzu C-RBA d a t a handl ing dev ice . A spheri-5

RP-18 column, l O O m m long and 4.6mm i n d iameter , was used

packed with 5clm p a r t i c l e s s i z e s . A mixture of

methanol/water ( 80:20 v/v ) with 1% a c e t i c ac id was

used as a mobile phase wi th f ixed flow rate

a t room temperature and w i t h a d e t e c t i o n wavelength of

230nm.

Der iva t ive s p e c t r a were measured in lcm qua r t z c e l l s

using a P e r k i n - E l m e r Lambda 5 UV-visible double beam

spectrophotometer. S u i t a b l e s e t t i n g s a r e : s l i t width

l . O m l m i n - ' ,

Znm, response time 5s, scan speed BOnmmin-' and mode d 'A

D4 = - d A4

Chromatographic analysis

TWO s e r i e s of ten s o l u t i o n s from O.Olmgml-' t o

( a c e t y l s a l i c y l i c acid ) and from 0.03 t o 0.3mgml-'

( chlormezanone ) were prepared f o r c a l i b r a t i o n curves .

The i n j e c t i o n s were repeated t h r e e times f o r each concen-

t r a t i o n .

A t t he same time t h e s tandard s o l u t i o n s of two subs tances

i n a mixture were prepared i n t h e same concen t r a t ions

as t h e analysed s o l u t i o n s . We compared t h e peaks a r e a .

The primary so lu t ion was prepared by d i l u t i n g 300mg of

a c e t y l s a l i c y l i c ac id and lOOmg of chlormezanone i n t h e

l O O m l mobile phase.

O.lmgml-'

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1696 IVANOVIC ET AL.

Ten t a b l e t s were weighed, powdered and d i l u t e d i n t h e

mobile phase. Af te r f i l t r a t i o n we prepared t h e same

concent ra t ion of t h e aforementioned analysed subs tances .

Spectrophotomtric analysis

A fou r th d e r i v a t i v e s p e c t r a f o r t h e s tandard mixture

s o l u t i o n s and t h e inves t iga t ed s o l u t i o n s was taken under

the same cond i t ions .

Ten mixture s o l u t i o n s of i nc reas ing concen t r a t ions of

a c e t y l s a l i c y l i c ac id ( from 0.02 t o 0.2mgnl-' )

decreas ing concen t r a t ions of chlormezanone ( from 0.2

t o 0.02mgml-' ) were then prepared and ana lysed .

The s tandard working so lu t ion was prepared by d i l u t i n g

150mg of a c e t y l s a l i c y l i c ac id and 50mg of chlormezanone

i n l O O m l of methanol w i t h add i t ion of 3% a c e t i c ac id .

This so lu t ion was d i l u t e d t o ob ta in concent ra t ions of

0 , l S and 0.05mgml-', r e spec t ive ly . Ten t a b l e t s were weighed and powered. 0.2182g of powder

was d i l u t e d in the volumetr ic f l a s k conta in ing l O O m l of

methanol. This s o l u t i o n was d i l u t e d t o ob ta in concentra-

t i o n s of 0 .15 and 0.05n1gml-~, r e s p e c t i v e l y .

and

RESULTS AND DISCUSSION

A l i n e a r c o r r e l a t i o n was obtained between the s u r f a c e of

t h e peaks and eoncen t r a t ions over t h e range 0 .03 - 0.3mgml-' f o r a c e t y l s a l i c y l i c a c i d , and 0 .01 -

f o r chlormezanone. The c o e f f i c i e n t is 0.9995 f o r each

l i n e a r r eg res s ion .

O.lmgml-'

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DETERMINATION OF ACETYLSALICYLIC ACID 1697

Y cg c;

Time (min)

Fig.1 Chromatogram of acetylsalicylic acid ( 1 ),

c = O.Gmgml-' and chlormezanone ( 2 ), c = O.OZmgml-i

Eluent: methanol/water ( 80 : 20 v/v )

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1698 IVANOVIC ET AL.

Table 1

Q u a n t i t a t i v e d e t e r m i n a t i o n of a c e t y l s a l i c y l i c a c i d and

chlormezanone i n Trancoges i c@ t a b l e t s

Acetylsalicylic a c i d

Met hod C h l ormezanone

amunt m U T 1 t Std C.V. masured found Dev. c % 3 C a r g ) C m g )

High-performance 300.00 300.00 2.11 2.10 -- 100.00 98.70 1.71 1.73 1 iqu i d

chromatography

0.99 1.00 98.93 1.66 1 .66 -- D e r i v a t i v e 300.00 298.74

spec t ropho tomet ry l o o ~ o o

n = 10

A c e t y l s a l i c y l i c a c i d is more p o l a r than chlormezanone

and is e l u a t e d f i r s t d u r i n g s e p a r a t i o n by l i q u i d

reversed-phase chromatography. These components were

s i m u l t a n e o u s l y de t e rmined i n TrancogesicQ t a b l e t s . The

excipient p r e s e n t d i d n o t i n t e r f e r e w i t h t h e de t e rmi -

n a t i o n of t h e s e two s u b s t a n c e s .

The r e s u l t s o b t a i n e d were a c c u r a t e and s a t i s f a c t o r y

( F i g . 1 . and Tab le 1. 1.

D e r i v a t i v e spec t ropho tomet ry has been shown t o be a

u s e f u l means of r e s o l v i n g o v e r l a p p i n g spectra and of

e l i m i n a t i n g m a t r i x i n t e r f e r e n c e i n t h e assay of a main

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DETERMINATION OF ACETYLSALICYLIC ACID 1699

s u b s t a n c e . I t i n c r e a s e s t h e f i n e s t r u c t u r e of t h e bands

obta ined by c l a s s i c a l UV-VIS spec t rophotometry .

A d e r i v a t i v e s p e c t r a has two advantages : r e s o l u t i o n is

much b e t t e r and t h i s r e s o l u t i o n i n c r e a s e s w i t h t h e

d e r i v a t i v e o r d e r . The ampl i tude , Dn, of t h e n- th

d e r i v a t i v e is i n v e r s e l y r e l a t e d t o t h e o r i g i n a l band

width, W , which is given i n t h e equation":

D" a W-"

T h i s is t h e reason why we have chosen t h e D, method. For

t h e mixture of two s u b s t a n c e s , A and B , it is recommended

t h a t a wavelength be chosen a t which t h e s i g n a l of A

is z e r o and t h e s i g n a l ob ta ined is dependent only on t h e

c o n c e n t r a t i o n of B ( " z e r o c r o s s i n g " method ) . I n o u r

d e t e r m i n a t i o n w e showed t h a t t h e " z e r o c r o s s i n g " method

is t h e method of c h o i c e .

A c e t y l s a l i c y l i c a c i d and chlornezanone o v e r l a p i n t h e z e r o

o r d e r spectra ( F i g . 2 . ), and i n t h e f i r s t , second and t h i r d

d e r i v a t i v e s p e c t r a , whi le i n t h e f o u r t h d e r i v a t i v e s p e c t r a it

is p o s s i b l e t o de te rmine s imul taneous ly t h e components i n a

mixture ( P i g . 3 . ) .

A c e t y l s a l i c y l i c a c i d is determined by t h e D, method a t

= 288.0nm ( z e r o p o i n t f o r chlormezanone ) and c h l o r -

mezanone is determined a t A = 263.71111 ( z e r o p o i n t f o r

a c e t y l s a l i c y l i c a c i d ) .

The c a l i b r a t i o n c u r v e s f o r two ser ies of t e n s o l u t i o n s

show a good l i n e a r i t y , wi th c o e f f i c i e n t s of r e g r e s s i o n

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1700 IVANOVIC ET AL.

A

0.12

Q06

I I \A I I \

I I I I I I

1

250 300 A nm

F i g . 2 Zero order spectra of ace ty l sa l i cy l i c acid ( - ) *

c = O.Olmgml-' and chlormezanone ( --- >. c = ~.lmgml-' i n methanol

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DETERMINATION OF ACETYLSALICYLIC ACID 1701

04

0.09

0.03

- 0.0:

- 0.0s

1 I I

260 300 A nm

Fig. 3 Fourth derivative spectra of acetylsalicylic acid

( - ), and chlormezanone ( --- 1, c o.lmgarl-'

in methanol

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1702 IVANOVIC ET AL.

of 0.9998 and 1.0028, respectively. The results obtained by

fourth derivative spectrophotometry are in agreement with

those obtained by HPLC and with the declared concentrations

( Table 1. ).

CONCLUSIONS

The authors propose two methods for simultaneous deter-

mination of acetylsalicylic acid and chlormezanone in

Tr ancoges ic" tablets . The results of both methods are accurate and repro-

ductive, but UV-derivative spectrophotometry is nore

rapid and simpler than HPLC.

ACKNOWLEDGEMENT

We thank the Winthrop laboratory for providing samples

and standard substances.

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DETERMINATION OF ACETYLSALICYLIC ACID 1703

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Received A p r i l 3 , 1992 Accepted May 2 7 , 1992

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