• S. Meena
  • S. M. Sandhya



Objective: Two simple, accurate and reproducible spectrophotometric methods have been developed and validated for simultaneous estimation of pyrimethamine and sulphadoxine in combined dosage form. Methods: Pyrimethamine shows maximum absorbance at 285.6 nm and sulphadoxine shows maximum absorbance at 273.4 nm. In dual wavelength method, two wavelengths were selected for each drug in a way so that the difference in absorbance is zero for another drug. Pyrimethamine shows equal absorbance at 278 and 295 nm, where the difference in absorbance was measured for determination of sulphadoxine. Similarly, difference in absorbance at 246 and 298 nm were measured for determination of pyrimethamine. For multicomponent mode method; the sampling wavelengths selected were 285.6 and 273.4 nm. Results: Linearity for detector response was observed in the concentration range of 3-18 µg/mL for pyrimethamine and 7-42 µg/mL for sulphadoxine for method I, 5-30 µg/mL for pyrimethamine and 10-60 µg/mL for sulphadoxine for method II respectively. Accuracy and precision studies were carried out and results were satisfactory. The proposed methods were validated as per ICH guidelines. Conclusion: The developed methods are simple, precise, rugged and economical. The utility of the methods has been demonstrated by analysis of commercially available formulations.


Key words: Pyrimethamine, Sulphadoxine, Dual wavelength method, Multicomponent analysis, Method validation.


1. The Indian Phamacopoeia, Vol. III, The Indian Pharmacopoeia Commission, 5th edn., India: NISCAIR Press, 2007, 1626.
2. United States Pharmacopoeia/The National Formulary (USP31/NF26), United States Pharmacopoeial Convention, Rockville, MD, 2008, 3295.
3. The Pharmcological basis of therapeutics, Goodman and Gilman’s, Joel. G. Hardman, Lee. E. Limbird, 1080-81.
4. Onah JO, Odeiani JE, Simultaneous spectrophotometric determination of sulphadoxine and pyrimethamine in pharmaceutical formulations. J Pharm Biomed Anal 2002;30 :851-857.
5. Green MD, Mount DL, Todd GD, Determination of sulphadoxine concentrations in whole blood using C18 solid phase extraction, sodium dodecyl sulfate and dimethyl amino cinnamaldehyde. Analyst 1995;120 :2625-2626.
6. AtemnKeng MA, Chimanuka B, Plaizier-Vercamman J, Quality evaluation of chloroquine, quinine, sulphadoxine-pyrimethamine and proguanil formulations sold on the market in East Congo DR. Clin Pharm Ther 2007;32 :123-132.
7. Berzas JJ, Castaneda G, Guzman FJ, Simultaneous determination of sulfamethazine and pyrimethamine by liquid chromatography. J Chromatography A 2000;870 :169-177.
8. Eljjaschewitsch J, Padberg J, Schurmann D, High performance liquid chromatographic determination of pyrimethamine, dapsone, monoacetyldapsone, sulphadoxine, and N-acetyl-sulfadoxine after rapid solid-phase extraction. Ther Drug Monit 1996;18 :592-597.
9. Astier H, Renard C, Cheminel V, Soarer S, Simultaneous determination of pyrimethamine and sulphadoxine in human plasma by high-performance liquid chromatography after automated liquid-solid extraction. J Chromatography B 1997;698 :217-223.
10. Bergquist Y, Eriksso M, Simultaneous determination of pyrimethamine and sulphadoxine in human plasma by high-performance liquid chromatography. Trans Royal Soc Trop Med Hyg 1985;79 :297-301.
11. Midsko C, High-performance liquid chromatographic assay of pyrimethamine, sulfadoxine and its N4-acetyl metabolite in serum and urine after ingestion of suldox. J Chromatography 1984;308 :217-227.
12. Edste M, Quantification of antimalarial drugs: I. Simultaneous measurement of sulphadoxine, N4-acetyl sulphadoxine and pyrimethamine in human plasma. J Chromatography 1984;305 :502-507.
13. Koesukwiwat U, Jayenta S, Leepipatpiboon N, Validation of a liquid chromatography-mass spectrometry multi-residue method for the simultaneous determination of sulfonamides, tetracyclines, and pyrimethamine in milk. J Chromatography A 2007;140 :147-156.
14. Suvarna I, Indravadan C, Bhagwat AM, Sundaresan M, Determination of sulfadoxine in human plasma using packed-column supercritical fluid chromatography. J Chromatography B 2001;757 :39-47.
15. Idowu OR, Dada OA, Simple and sensitive spectrofluorimetric determination of pyrimethamine in biological fluids. Anal Chim Acta 1985;169 :361-365.
16. ICH Q2 (R1), Validation of Analytical Procedures: Text and Methodology, CPMP/ICH/381/95, Geneva, 1995, 1-15.
628 Views | 455 Downloads
How to Cite
Meena, S., and S. M. Sandhya. “VALIDATED SPECTROPHOTOMETRIC METHODS FOR SIMULTANEOUS ANALYSIS OF PYRIMETHAMINE AND SULPHADOXINE IN PHARMACEUTICAL DOSAGE FORMS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 6, no. 7, Aug. 2013, pp. 121-3,