GHTMDD - 249

Bulletin of the Chemists and Technologists of Macedonia, Vo\.14, No. 1, p. 39 - 41 (1995)

ISSN 0350 - 0136

Received: November 1st, 1993 UDC 615. 2 : 543. 544

Short communication

DETERMINATION OF PROPYPHENAZONE, PARACETAMOL, CAFFEINEAND CODEINE PHOSPHATE WITH THIN LAYER CHROMATOGRAPHY

Aneta Dimitrovskat, Suzana Trajkovic-Jolevskat, Amalija Nancovskat and Miroslava Ilievska21Institute for Examinations and Control of Dmgs, Faculty of Pharmacy.

The "Sv. Kiril i Metodij" University, 91000 Skopje, Macedonia.2A.D. Alkaloid, 91000 Skopje, Macedonia

The conditions have been examined for the determination of propyphenazone, paracetamol, caffeine and codeinephosphate in Caffetin tablets with preparative thin layer chromatography. The separation of propyphenazone,paracetamol and caffeine was performed by use of a mobile phase chloroform-acetone-ammonium hydroxide (25%) involume rations 8:2:0.1. Codeine phosphate was separated from the other componentswith chloroform-ethanol in volumeration 8:2, asa mobile phase. The location of spots on the chromatogram was detected with UV lamp on 254 nm. Everycomponent was eluted from the adsorbent with a solution of 0.1 mollL hydrochloric acid in ultrasonic bath for 5 min.The solutions were filtered and absorbency was measured at 240 nm for propyphenazone, 245 nm for paracetamol, 273nm for caffeine and 285 nm for codeine phosphate. The results show that the applied method is fast, reproducible andsuitable for routine analysis.

Key words: propyphenazone; paracetamol; caffeine; codeine phosphate; thin layer chromatography

INTRODUCTION

Caffetin tablets belong to the group of nonnarcotic anal go-anti pyre tics. They are a combinationof four components: propyphenazone, paracetamol,caffeine and codeine phosphate. In this combinationcaffeine and codeine phosphate increase the analgesiceffect of the paracetamol and the propyphenazone,synergisticly.

The literature information relate mostly to thedetermination of those active components in biologi-cal materials using gas chromatography (GC) [1-4],high performance liquid chromatography (HPLC)[5-8] and gas chromatography in combination withmass spectroscopy [9].

In the literature, there are informations on theseparation and the determination of caffeine from

some analgo-antipyretic mixtures. with thin layer.

chromatography [10.'11, 12]. We didn't find literatureinformation on the determination ofpropyphenazone,paracetamol, caffeine and codeine phosphate whenthey are in the same mixture. In aqueous-acid solutionsthose substances have close absorption maximum inUV-region [13] and their determination directly fromthe mixture is impossible. Our exam,inations werebased on UV-spectrophotometric determination of,components in dosaged pharmaceutical forms, Caf-fetin tablets, after their separation on thin layer andelution from the adsorbent.

Examined sample

EXPERIMENTAL

Standards

Caffetin tablets - Alkaioid, SkopjeIt is declared:

ProphyphenazoneParacetamol

Caffeine

Codeine phosphate

0.210 g

0.250 g

0.050 g

0.010 g

Propyphenazone, Paracetamol, Caffeine andCodeine phosphate - Alkaloid, Skopje

Procedure for determination of propyphenazone,paracetamol and caffeine

The average mass of twenty pulverized tabletswas measured, dissolved with ethanol in 50 ml volu-metric flask and the solution was filtered. The calcu-

40 A. Dimitrovska, S. Trajkovska-Jolevska (et al.)

lated concentration ofpropyphenazone in the solutionwas 4.2 mg/ml, paracetamol 5 mg/ml and caffeine 1mg/ml.

The concentrations of the substances in the

sample were equal to those of the standards in ethanolsolutions.

25 ,ul from solutions of the standards and thesample were applied on plates of Sillica gel 60 F254'0.250 mm, Merck.

The chromatogram was developed by use of amobile phase chloroform-acetone-ammonium hy-droxide (25%) in volume rations 8: 2: 0.1.

Procedure for determination of codeine phosphate

The average mass of twenty pulverized tabletswas measured, dissolved with 1.5 ml hot distilled waterin '10 ml volumetric flask, ethanol was added till themark and the solution was filtered. The calculated

concentration of codeine phosphate in solution was 1mg/ml.

The codeine phosphate standard was prepared inthe saIne way and with equal concentration as thesample.

200,u1 from the solution of the standard and thesample was applied on plates of Sillica gel 60 F254'0.250 mm, Merck. The chromatogram was developedby use of a mobile phase chloroform-ethanol involume ration 8 : 2.

The spots of chromatogram were detected withUV-lamp on 254 nm. The marked spots of separated

compolents were eluted from adsorbent with solutionof 0.1 mol/! hydrochloric acid, using 20 ml for prohy-phenazone, 25 ml for paracetamol, 5 ml for caffeineand 5 ml for codeine phosphate. The solutions werefiltered and absorbency was measured at 240 nm wavelength for propyphenazone, 245 nm for paracetamol,273 nm for caffeine and 285 nm for codeine phosphate,on spectrophotometer Pye Unicam 8600 UVNIS.

Calibration diagram for propyphenazone,paracetamol and caffeine

From the standard solution of propyphenazone,paracetamol and caffeine with concentrations of 5mg/ml, applied were 5,10, 15,20 and 25,u1on plates ofSillica gel 60 F254'0.250mm,Merck.The componentswere eluted from the adsorbent with 10 ml solution of0.1 mol/! hydrochloric acid. The solution concentra-tions of components were: 2.5,5,7.5,10 and 12.5,ug/ml.

Calibration diagram for codeine phosphate

From the standard solution of codeine phosphatewith concentration of 2 mg/ml, applied were 50, 100,150,200 and 250,u1on plates ofSillica gel 60 F254'0.250mm, Merck. Codeine phosphate was eluted from theadsorbent with 5 ml solution of 0.1 mol/! hydrochloricacid. The solution concentrations of codeine phos-phate were: 20,40,60, 80 and 100,ug/ml.

RESULTS AND DISCUSSION

Several mobile phases with related Rf values arecited in literature [14] on the separate determinationofpropyphenazone, paracetamol, caffeine and codeinephosphate. The proposed mobile phases didn't giveoptimal separation of components, for their furtherquantitative elution from the adsorbent. The bestseparation of components was obtained with mobilephase chloroform-acetone in volume ration 8 : 2

(propyphenazone Rf = 61; caffeine Rf = 23; para-cetamol Rf = 19; and codeine phosphate stay on thestart line). More suitable pH value for better separa-tion of components [15] was obtained by addingammonium hydroxide on the mobile phase chloroform

-acetone, cited in the literature. TheRf values with theused mobile phase chloroform-acetone-ammoniumhydroxide (25%) in volume rations 8: 2 : 0.1 were forpropyphenazone 89, for caffeine 47, for paracetamol19 and for codeine phosphate 6. Because of low Rf '

value, codeine phosphate was separated from the othercomponents by use of the mobile phase chloroform-ethanol in volume ration 8 : 2 (Rf = 35). In the

literature [14) on the determination of codeine phos-phate with thin layer chromatography chloroform-methanol as a mobile phase is suggested, but methanolwas changed with ethanol because solutions of thesample and the standards were prepared with ethanol.For determination of codeine phosphate, the largervolume from the solution of sample was applied,because of its low molar absorption [13] and smallcontents in Caffetin tablet.

Linear dependence between absorption and con-centration of active component was established in therange of concentration 2.5 - 12.5 ,ug/ml for propy-phenazone (r = 0.9994), paracetamol (r = 0.9996) andcaffeine(r = 0.9991)and in the range of concentration20-100,ug/ml for codeine phosphate (r = 0.9989).

The results of UV spectrophotometric deter-mination of propyphenazone, paracetamol, caffeineand codeine phosphate from Caffetin tablets, aftertheir elution from adsorbent, are presented in '!able I.

The values of RSD show that the suggested

method can be used for routine analysis [16r

BuII.Chem.TechnoI.Macedonia,14, 1, p.39-41(1995)

Determination of propyhenazone, paracetamo~ caffeine 41

Ta b Ie I

Results from the determination of propyphenazone, paracetamol, caffeine and codeine phosphate from Caffedn tablets

CONCLUSION.

The suggested method allows UV spectrophoto-metric determination of propyphenazone, para-cetamol, caffeine and codeine phosphate from Caffetin

tablets after their separation on thin layer and elutionfrom adsorbent. The method is fast, reproducible andsuitable for routine analysis.

REFERENCES

[1] I. O. Bradbrook et al.,J. Chromat., 163,118 (1979).

[2] G. R. Nakamura and E. L. Way,Analyt. Chem., 47,775 (1975).

[3] P. O. Edlund,J. Chromat., 206,109 (1981).[4] E. Kaa,J. Chromat., 221,414 (1980).

[5] O. B. Haughey et aI.,J. Chromat., 229,387 (1982).

[6] S. E. O'Connel and F. J. Zurzula,J. Pharm Sci., 73,1009 (1984).

[7] V. Nitsche and H. Mascher,J. Pharm. Sci., 73,1556 (1984).

[8] J. N. Buskin et aI., J. Chromat., 230, 443 (1982).

[9] N. B. Wu Chen et aI., J. Analyt. toxicol., 6,231 (1982).

[10] O. Radulovic, M. Pokrajac, M. Primorac,Arh. farm., 34 (2), 73(1973).

[11] O. Radulovic, M. Stanojcic,Arh. farm, 6, 321 (1973).

[12] O. Radulovic, Z. Blagojevic, Z. Sreckovic, Arh. farm, 4, 217(1975).

[13] E. G. C. Clarke, Clarke's Isolation and Identification of Dmgs,Second edition, Part 2, The Pharmaceutical Press, London,1986, pp. 420, 490, 849 and 939.

[14] E. G. C. Clarke, Clarke's Isolation and Identification of Dmgs,Second Edition, Part 1, A C. Moffat, Thin layer chromato-graphy, p. 160.

[15] A H. Stead et aI., Ana(yst, London, 107, 1106 (1982).

[16] I. M. Kolthoff, P. J. Elving, Treatise on Analytical Chemistry, 2ndEdition, Part 1, Vol. 1, Theory and Practice, John WHey andSons, New York, 1978, p. 191.

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Examined componentsOeclared mass Found mass SO" RSO

per tablet / g per tablet / g gltablet %

Propyphenazone 0.210 0.2082 3.1'10-3 1.48

Paracetamol 0.250 0.2485 2.6 '10-3 1.05

Caffeine 0.050 0.0493 7.9 '10-4 1.61

Codeine phosphate0.010 0.0098 1.7 '10-4 1.76

-"n =10