Phosphate and Ephedrine Hydrochloride by Derivative UV Spectrophotometry

FABAD J. Pharm. Sci., 21, 55-60, 1996

RESEARCH ARTICLES I BiLiMSEL ARAŞTIRMALAR

Analysis of Syrups Containing

Chlorpheniramine Maleate, Codeine

Phosphate and Ephedrine Hydrochloride by Derivative UV Spectrophotometry

Filiz Sayın*, Sedef Kır*, Aytekin Temizer*

Analysis of Syrups Containing Chlorphenirarnine Maleate, Codeine Phosphate and Ephedrine Hydroclıloride by

Derivative UV Spectrophoton1et1y ..

Abstract : A method was presented for the deterrnination of

clılorpheniraınine ınaleate (1), codeine phosphate (11) and ephedrine lıydrochloride (lll) in a mixture of thenı without any separation step. The coınpounds 1 and il can be de-

ternıined sinıultaneously by using the second derivative Ull

spectrophotonıetry. The conıpound fil can be deternıined by the saıne nıethod after the oxidation with sodiuın rneta- periodate. The linear ranges of concentrations far the analy- sis of!, Il and III have been found as 0.001-0.08, 0.001-0.4 and 0.005-1.80 mg!nıl, respectively. The nıethod developed was applied to pharmaceutical fonns of these con1pounds.

The percent recoveries 'rverefound as 101%for1, 100 %/or II and 100 % for Ill.

Keywords : Derivative UV spectrophotonıefl)', chlor- pheniramine nıaleate, codeine phosphate and·ephedrine hydrochloride,phar- maceutical preparations.

Introduction

Chlorpheniramine maleate (!), codeine phosphate (il) and ephedrine hydrochloride (III) are frequently pre- scribed as a sedative cough mixture. The different methods proposed for I in mixture with Il phenylephrine hydrochloride and acetarninophen TLCI, with phenylpropanolamine hydrochloride by GLC2,

and determinations by Ion-Pair HPLC^3, with III and guaiacolsulfonate potassium using colorimetric method⁴ have been reported. Few applications of derivative spectrometric technique to the determination of !, using second derivative

Klorfeniramin Maleat, Kodein J?osfat ve Efedrin Hidroklorür İçeren Şurupların Türevsel UV

Spektrofoto1netrisi ile Analizi

Özet: Klorfeniranıin ınaleat (!), kodein fosfat (il) ve efedrin hidroklonir'ün (/il) karışunlardan ayırına işleıni ya-

pılnıadan tayini için bir yönten1 geliştirilnıiştir. Bileşik J ve II ikinci türev UV spektrofotoınetrisi ile tayin edihniştir. Bi-

leşik 11! ise sodyunı nıetaperiodat ile yükseltgen.dikten sonra

aynı yöntem.le tayin edilmiştir. 1, il ve l/l'ün anaUzi için de-

rişimin doğrusal olduğu aralık sırasryla 0.001-0.08, 0.001- 0.4 ve 0.005-1.80 nıghnL olarak bulunnıuştur. Geliştirilen yöntenı bu bileşikleri içeren şurup farmasötik preparat/ara

uygulanmıştır. Yüzde geri kazanım 1, 11ve111 için sıra ile%

101, %100ve%100 olarak bulunınuştur.

Anahtar kelimeler : Türev UV Spi!ktrofotornetrisi, klor- feniramin maleat, kodein fosfat ve

efedrin lıidroklorür, şun1p.

pectrophotomeıryS, with pseudoephedrine hydro- chloride assay first derivative UV spectrophotometry^6, with dextrometorphane hydrobromide, and pseudo- ephedrine hydrobromide using second derivative diode-array spectrophotometry7, with pyrilamine maleate, and phenylpropylamine hydrochloride as- say difference spectrophotometry8 and the com- pound I was deterrnined by polarographic tech- nique9. Recently; it has been shown that the application of derivative techniques to spectro- photometry is very useful in resolving spectral over- lap and in cancelling irrelevant absorption from the secondary sample ingredientsrn

*

Presented at 3th Intemational Syınposiuın ofPharınaceutical Sciences, 15-18 June 1993, Ankara, T\JRKEY.

Sayın, Kır, Tenıizer

The present paper deals with the determination of the cornpounds 1, il and ili in three-componcnt rnixture using derivative SpL'Clrophotornctric teclmique.

Material and Methods Instrument

Shimadzu 160-A Model ^UV spectrophotorneter was used. ^it is a rnicrocontrolled double-bcam recording spectrophotometer. The spectra of test and reference solutions were rccorded in 1.000 cm quartz cells over the range 220 to 400 nm. Suitable settings are;

scan speed: 2400 nrn/rnin, rnode: D₂ (second deri- vative)

=

Reagents

All reagents and chernicals used were of analytical reagent gradc.

1. Phosphate buffer pH 7.00: Mix 295.4 rnL of 0.1 N sodiurn hydroxide and 250 rnL 0.2 M rnonobasic po- tassium phosphate in 1 L calibrated flask and corn- plete to volurnc with water.

2. 0.2 M sodium metaperiodate solution : Dissolve 10.695 g of sodiuin rnetaperiodate in 250 mL water.

Store in a dark glass bottle.

Materials

1. Chlorpheniramine maleate (SIGMA) 2. Codeine phosphate (SJGMA) 3. Ephedrine hydrochloride (SlGMA)

Dephedrin ^® syrup : Labelled to contain 20 mg !, 150 mg il and 150 mg llI per 100 mL syrup.

Codis ® syrup : Labelled to contain 20 mg !, 200 mg il and 240 mg III per 100 mL syrup.

Corex ® syrup : Labelled to contain 100 mg !, 250 mg II, and 125 mg III per 100 mL syrup

in order to prepare standard solutions, 20 mg of !, 200 mg of II or 150 mg of 11I were dissolved in 100 rnL 0.1 N HCl. Thcy were prepared separately and cornbinations of them.

Experimen!al Data

Prepara!ion ot syrup sample

a) For 1 and il : Transfer accurately 20 mL aliquot of syrup to separatory funnel. Add 50 rnL water and 5 mL 1 N HC! and extract far 10 min with 20 mL di- ethylether saturated with 1 N HCI. Wash the etheri- al layer with three ¹⁰ mL portions of 0.1 N HCl.

Madc basic the combined washings and aqueous layer with 15 mL 1 N NaOH and extract with four 15 mL portions of chloroforrn. Collect the organic extracts and rc-extracted with six mL portions of 0.1 N HCl. Filter each acidic extract through a filler papcr moistured with 0.1 N HC! and collect the filtrate in a 100 rnL calibrated flask and make up to volume with 0.1 N HCl.

b) For 111: Transfer accurately 10 mL aliquot of syrup to separatory funnel. Made basic with 5 mL 5 N NaOH an add 15 mL saturated Nacı solutions. Ex- tract with three 20 mL portions of chloroform. Col- lect the chlorformic extracts in another separatory funnel and re-extract with three 25 mL portions of 0.1 N HCl and collect the acidic extracts in a 100 mL calibrated flask and complete to volume with 0.1 N HCI.

General Procedures

a) For I : Transfer from prepared standard solutions containing I or 25 mL syrup sample prepared as ex- plained above into different calibrated flask .and corrlplete the volurne 50 mL with 0.1 N HCl. D_{2 -}UV spectra were recorded against 0.1 N HC! and peak amplitude at 288.2 nm was measured.

b) For il: Transfer from prepared standard solutions containing il or 25 mL syrup sample prepared as ex- plained above into different calibrated flask and complete the volume 50 mL with 0.1 N NaOH. ^Dı-UV spectra were recorded against 0.1 N NaOH and peak amplitude at 292.6 nrn was rneasured.

c) Far III : Transfer from prepared standard solutions containing III or 1 rnL syrup sample prepared as cxplained above into separatory funnel. After ad- ditions of 0.2 M sodium metaperiodate solution and 2 N NaOH, pH was adjusted to 7.50. The solution was mixed far 10 min. Then, 1 N HC! and n-hexane

FABAD J. Pharnı. Sci., 21, 55-60, 1996

were added and mixed lhorougly for 5 min. The hexane layer was filtered and complctc the volume 25 mL with n-hexanc. Dz-UV spectra of the extracts were recorded against n-hcxane. Peak amplitnde at 255.2 nm was measurcd.

Results and Discussion

in this study, the nature of the solutions in which active substances were dissolved, degree of de- rivatives, the range of wavelength, "N" value (N = 3,

!ıA.= 10.5) and the thickness of the celi were the basic parameters.

Figures 1 a, b and c show the D_{2 -}UV spectra of 1, il and III in 0.1 N HCI, respectively. it was observed that there is no effect of il and il! at 288.2 nm. Ac- cordingly, the result was obtained when direct measurement of 1 using Amax D₂ at this analytical wavelength.

Figures 2 a, b and c show the D_{2 -}UV spectra of il, I and llI 0.1 N NaOH, respectively. it was observed that !here is no effect of 1 and III at 292.6 nm. TI1is wavelength was used for the quantitative de- termination of il in mixture with 1 and Ill.

III exhibits very low absorption in the UV region. in addition, the absorption rnaxima of III are over- laped by the absorption spectra of the other coex- isting cornponents. Oxidation of 1II with sodiurn metaperiodale as described in experirnental part on forrnation of benzaldehyde which increase the ab- sorptivity of III ^(11).

Figures 3 a, b and c show the D_{2 -}UV spectra of 111, 1 and il in n-hexane, respectively. So the application of sccond derivative technique to the oxidized rnixtnre at 255.2 nrn was used to analysc III in the mixtnre of I and il.

Table 1 has shown that the data of the graphs ob- tained by plotting the values of D₂ lor 1, il and ili (after oxidation) under the given experirnental con- ditions at the chosen wavelengths against the concen- tration are withiı1 a considerable interval. Cali- bration curves for I, il and 111 are shown in Figure 4- 6, respectively. The coefficient of determination and regression lor 1, il and III were determined frorn the experimental dala within working range

by linear regression analysis. TI1c deviation from the linearity was 'found !o be insignificant (95 %) after the preparation of synthetic mixtures of 1, il and III.

These mixtnres were analysed using spectro- photometric method following the extraction pro- ccdurcs. Then, percent recoveries of thc method vvas calculated by comparing the amount of compound in synthetic mixtures with the results obtained (Table 2). The proposed method has been applied to the simultaneous determination of the above men- tioned mixtnre. in cornrncrcial syrups (Table 3). The re- sults obtained have been compared with the methods registered in the Turkish Pharrnacopeia (1974), such as spectrophotometry for !, and titrirnetry for il and ili. No difference has been observed between !he 1, il and III <lata. The results laken by two different techniques lor Dephedrin ® syrup as an exarnple, were compared as seen on Table 4.

Table 1. The determincd parameters for calibration curves of 1, II and ili

Compound Solvenı Selected Conc.range lntercept Slope De ter.

(A)(nm) (mg/ml) (a) (b) cool! (i2)

0.1 N HCI 288.2 0.001-0.08 -9.37xl 0-⁴ 2.25 1.00 0.1 N NaOH 292.6 0.001-0.40 1.25x10'³ 0.723 1.00 111 n-Hexane 255.2 0.005-1.80 0.0111 0.539 1.00

Table 2. The results of percent recovery of standard mixtnres of !, il and 111

Compound Added Pound Recovery

(mg/10mL) (mg/10mL) %

0.50 0.50 100

1.00 1.03 103

1.50 1.50 100

2.00 2.03 101

5.00 5.01 100

il 10.00 10.0 100

15.00 15.0 100

20.00 20.0 100

10.00 10.0 100

III 15.00 15.0 100

20.00 20.0 99.9

25.00 25.0 100

Saı;1n, Kır, Temizer

Table 3. Assay results of!, ll and ili in commercial syrups.

Pharmaceutical preparations

Dephedrin®

Codis®

Corex®

2 mg/10 ml X=2.01±0.01 S=0.02 V=1o/o CI= 1.98·2.04

2 mg/10 ml X=2.00±0.01 S=0.02 V=1°/o CI= 1.97·2.03

10mg/10 ml X=10.0±0.0 S=0.1 V=0.7%

Cl=9.9-10.1

il 111

15 mg/10 ml 15 mg/10 ml X=15.0±0.0 X=15.0±0.0

S=1.2 S=O.O

V=7.9% V=0.27%

Cl=13.7·16.3 Cl=15.0·15.1

20mg/10 ml 24mg/10 ml X=20.0±0.0 X=24.0±0.0

S=0.1 S=0.1

V=0.35% V=0.25%

Cl=19.9·20.1 Cl=24.0·24.1

25mg/10 ml 12.SOmg/1 O ml X=24.9±0.0 X=12.5±0.0

S=0.1 S=O.O

V=0.44% V=0.32%

Cl=24.8-25.0 Cl=12.4-12.6

x = mean ± standard error, S ⁼ standard deviation, V = variation co- efficient, CI = confidence intervals (95%)

Table 4. Statistical evaluation of obtained <lata from developed Dz-UV technique and comparison with pharmacopeia technique far Dephedrin® syrup.

Compound Statistical

o,-uv

values technique technique (mg/ml) (mg/ml)

Spectrophotometric

n 6 6

x 2.01 1.97

s

V(Six.100) 1 1.01

cı 1.98-2.04 1.97-2.00

Titrimetric

n 6 6

il x 15.0 14.5

s

V(S/x.100) 7.9 1.30

cı 13.7-16.3 14.4-14.8

Titrimetric

n 6 6

111 x 15.0 14.6

s o.o

V(S/x.100) 0.27 1.78

cı 15.0-15.1 14.3-14.8

n = number of sample, x = mean, S = standard deviation, V(S/x.100) = variation cooficent, CI = confidence intervals (95%)

~il. lll2A

f\ \/

.; -"1'~ ~

.ı:.1ı:ıı .... )

-e .O.S:A

:?·Hı.O -· 20.0(Hl1/0[V.> ^{., eo. o}

""

.,.,;ı. lr)3A

Code!ne phosphat.e

" . ~ l.)

i\ ^\

^.

_.,,.. ol1.'. > .

\l\ı

-O, 113A

""

240.0 2&.€HHM/Ol-V.) <100 .ı:ı

.,, .u ı;;

Ephcd~e hydrochlortde

iL

- ^{- .} ..

~ lı/\1

. . -

:ı -~'4S . 1 '

.=.; ;)t" . ' 1

• \İ. li2A

""

:;o .. p) .D. 20.0\llH,.OlV.> 400.U

Figure 1. Dz-UV spectra ofa) 0.2 mg/mL I, b) 1 mg/mL II and c) 4 mg/mL IIl in 0.1 N HO.

•Q .011iA

CÔdeine phı::ısphate

·~

~. ~Z.o)

ı\

(il./ fi ı ~. ~

\/

-Q. Q,;;ı

z:..:Q. Q ... ^{Ztl. Q{ ı.Ul/D} { '·' ._) •IQQ .(l ^Ut!

+O. l SA -

-~

r A /Öl·~.) -

-0,20A

""

2<1'CI .0 Z0.0(Hl1,.0IV.) <100 .o

+0.01A

\

...

-

(1. Oll:S

~ .:ı..-o ı •J.'

-0 .ı:ızı=ı

;~" " ^{- - - .. ~,.n111 '} """H~

Figure 2. DrUV spectra ofa) 1 mg/mL II, b) 0.2 mg/mL l and c) 4 rng/rnL II1 in 0.1 N NaOH.

FABAD J. Pharm. Sci., 21, 55-60, 1996

.. o. 15Fı

Ephedrlne: hydrochlorlde

•

ı). •'.•~O ,\

,;,,rcıı~•.>

\j'

'

\

-O. 1 SA

2.20 .o ^{2:0 .0( ıttt~·tı [ v.) -ı (•.'.• ı_ı~}

+•). ı).SA

IA

Chlorpheniramine maleate

"i~ \

•) . •)(El

1

( ıı;o ıu.)

;

/\

- _--

1 ı

'v - - ^{. ..}

-u. o.ı,;

220 .o 20.U.:tlN,.~•llJ,) -1 (•(•.,) ^ı.:~

... a.o·t-A- · ^{-· -·} -

Codelne phosphate ·

o. 01):5 ,r,__ ~ f\ A

.

(\

( ... o [ v.)

vv _,)

. _{.- 'V v}

-8.81A

'"

220.0 213.8(H!1/0IU.) -100 ...

Figure 3. DrUV spectra ofa) 4 mg/ml Hl, b) 0.2 mg/mL I and c) 1 mg/mL II in n-hexane.

0.25

.

0.2

•

0.15

~ •

~

0.05

.

0.025 0.05 0.075 0.\

Coııt~nıration (mglml)

y"' 2.25 x + -9.37xıo-4

r2 = 1.00 ; p < 0.05

Figure 4. Calibnıtion curve of l at 288. 2 nrn.

0.3

0.2

0.\

~·

•

.

•

•

•

O.\

•

•

0.2 0.3

Conteııtnıtion (mg/ml)

y= o.723x+ ı.25xıo·3 r2= ı.oo: p<O.os

Figure 5. Calibration curvc of II at 292.6 nm.

•

0.75

•

;;. ~

"

0.5

0.25 •

I

0.5

Cımccntr&tion {mg/ml)

Y= 0.539x+ 0.0111 r2= ı.oo : p<O.os

•

Figure 6. Calibration crnve of III at 255.2 nm.

0.4

•

•

'

Sayın, Kır, Tenıizer

The possibility of the intcrfcring constituents in syr- up must not be ovcrlookcd. Thcrefore, the standard addition technique "\vas used to evaluate the ac- curacy of the proposcd method far such samples and to test far interference from the syrup constitu- ents.

it has been concludcd, that thc method developed, namely derivative UV spectrophotometry is sensi- tive, accurate, precise and reproducible.

References

1. Al-Kaysi, H. N., Salen1, M.S., "Simultaneous Quanti- tative Detern1ination of Codeine Phosphate, Chlor- phenireamine Maleate, Phenylephrine Hydro- chloride and Acctan1iı1ophcn in Pharmaceutical Dos- age Forms Using Thin Layer Chromatography Densitometry'", Ana/. Lett., 19 (7-8), 915-924, 1986.

2. Madsen, R. E., Magin, D. F., ''Sin1ultaneous Quanti- tative CLC Determination of Chlorphenireamine Ma- lcate and Phenylpropanolamine Hydrochloride in a Cold Tablet Preparation'", J. Plu:ırnı. Sci., 65(6), 924- 925, 1976.

3. Greco, C.T., "Ion-Pair High Performance Liquid

Chroınatographic Detern1ination of Chlor- pheniramine Maleate in Cough Cold Mixtures", Drug Dev. Jnt. Plıarnı., 10, (1), 19-30, 1984.

4. Gupta, V. D., Lara, A. ). L., "Colorimetric De- terminations of Chlorphenirarnine Maleate, Ephedrine Hydrochloride, and Guaiacolsufonate Potassium in a Cough Syrup'', J. Plıarnı. Sci., 64 (12), 2001-2002, 1975.

5. Leung1 C. P., Law, C. K., "Dctcrn1ination of Chlor- pheniramine Maleate in Tablets by Second - De- rivative Absorption Spectrophotometry", A11alyst, 114 (2), 241-242, 1989.

6. Hoovcr, J. M., Soltero, R. A., Bansal, P. C., "Analysis of Multicomponent Forn11tlations Containing Pseudo- ephedrine Hydrochloride and Chlorpheniraınine

Maleate U sing First - Deri va ti ve Spectroscopy on a Diode - Array Spectrophoton1etcr", ]. Pharın. Sci., 76 (3), 242-244, 1987.

7. Murtha,]. L., Julian, TN., Radebaugh, G. W., "Simul- tancous Determination of Pseudoephedrine Hydro- chloride, Chlorpheniramine Maleate, and Dextro- 111ethorpha·n Hydrobromide by Second - Derivative Photodiodc Array Spectroscopy", ] . Pharnı. Sci., 77 (8), 715-718, 1988.

8. Tan, H. S. L, Salvador, G. C., "Assay of Mixtures of Chlorpheniran1ine Maleate, Pyrilamine Maleate and

Phenylpropanolaınine Hydrochloride in Cold - Aller- gy Tablets by Difference Spectrophoton1etry", Annl.

Chinı. Acta., 188, 295-300, 1986.

9. jacobsen, E., Hogberg, K., '"Polarographic Dc-

terınination of Chlorpheniramine Maleate in Pharına­

ceuticals", Aııal. Clıiııı. Acta ., 71, 157-163, 197 4.

10. Talsky, G., Mayring, L., Kreuzer, H., "High- Resolution, Higer-Order UV /VIS Derivative Spectro- photomctry", Angew. Clıenı. lııt. Ed. Eııgl., 17 (11), 785- 799, 1978.

1 L Chafetz, L., "Spccificity of Spectrophotometric De- termination of Ephedrine and Other Phe- nalkanolaminc Drugs as Benzaldehydes after Period- ate Oxidation'", J. Plıarnı. Sci., 60, 291-294, 1971.