• Ramakrishna Veni Pokala Department of Applied Sciences and Humanities, Sasi Institute of Technology and Engineering, Tadepalligudem 534101, Andhra Pradesh, India
  • Kusuma Kumari Department of Chemistry, Andhra Christian College,Guntur 522002, Andhra Pradesh, India
  • Hari Babu Bollikola Department of Chemistry, Acharya Nagarjuna University, Nagarjuna nagar, Guntur-522510, Andhra Pradesh, India


Objective: A new, simple, sensitive and economical UV spectrophotometric method was developed for the simultaneous analysis of Sulfadiazine [SDA] and Trimethoprim [TMP] in pharmaceutical formulations.

Methods: This UV method was developed with methanol as solvent. The wavelengths selected for analysis in the present method were 265 nm for TMP and 289 nm for SDA. Teccomp UV-2301 double beam UV/Vis spectrophotometer was used to carry out spectral analysis and the data was recorded by Hitachi software.

Results: Linearity was found to be within the concentration range of 2-9 µg/ml TMP and 9.08-41 µg/ml of SDA. Accuracy of the method was determined by recovery studies. Percentage recovery was found to be 98.20-99.25 for TMP with a % RSD of 0.338, 0.506 and 0.510 for three spiked levels. % RSD was found to be 0.229 and 0.380; 0.212 and 0.328 for SDA, TMP in intra and inter-day precision respectively. The % RSD value in ruggedness was found to be 0.440 for SDA and 0.569 for TMP.

Conclusion: The advantages of this method for analytical purposes lie in the rapid determination, its cost-effectiveness, easy preparation of the sample, good reproducibility. In addition to this, the present method can be recommended for simultaneous determination of SDA and TMP in routine quality control analysis in combined drug formulations.

Keywords: Sulfadiazine, Trimethoprim, UV-Method and Methanol


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1. De Araujo MV, Vieira EK, Silva Lazaro G, Conegero LS, Almeida LE, Barreto LS, et al. Sulfadiazine/Hydroxypropyl-beta-cyclodextrin host-guest system: characterization, phase-solubility and molecular modeling. Bioorg Med Chem 2008;16:5788-94.
2. Iliades P, Meshnick SR, Macreadie IG. Dihydropteroate synthase mutations in Pneumocystis Jiroveciican affect sulfamethoxazole resistance in a Saccharomyces Cerevisiae model. Antimicrob Agents Chemother 2005;49:741-8.
3. Prabhu V, Lui H, King J. Arabidopsis dihydropteroate synthase: general properties and inhibition by reaction product and sulfonamides. Phytochemistry 1997;45:23-7.
4. "WHO model list of essential medicines" World Health Organization October, Retrieved; 2014.
5. Rossi S. The Australian Medicines Handbook Unit Trust; 2013.
6. Vandana K, Arun Kumar D, Umadevi K, Shiva KT, Harika L, Kishant Kumar P. Method development and validation of sulphadiazine in bulk and pharmaceutical dosage form by UV-spectrophotometric method. Int J Pharm Biol Arch 2011;2:1167-71.
7. Salim AM, Haseeb YSZ. Spectrophotometric determination of sulfadiazine via diazotization and coupling reaction-application to pharmaceutical preparations. Raf J Sci 2013;24:61-73.
8. Goran S, Elizabeta DS, Marija S, Romel V. Optimization, validation and application of UV-Vis spectrophotometric-colourimetric methods for determination of trimethoprim in different medicinal products. Mac Vet Rev 2016;39:65-76.
9. Shamsa F, Amani L. Determination of sulfamethoxazole and trimethoprim in pharmaceuticals by visible and UV spectrophotometry. Iran J Pharm Res 2006;1:31-6.
10. Behzadian NG, Rezaee A, Kebriaeezadeh A. High-performance liquid chromatographic determination of trimethoprim in mouse liver. Pharm Pharmacol Commun 1998;4:439-41.
11. Nordholm L, Dalgard LA. Assay of trimethoprim in plasma and urine by high-performance liquid chromatography. J Chromatogr 1982;233:427-31.
12. Springolo V, Coppi G. HPLC determination of tetroxoprim and sulphadiazine in pharmaceutical dosage forms and in biological fluids. J Pharm Biomed Anal 1989;7:57-65.
13. Goulas V, Anisimova AT, Angastinioti MC, Tzamaloukas O. A rapid HPLC method for the determination of sulphonamides and trimethoprim in feed premixes. J Anim Feed Sci 2014;23:185–9.
14. Mashhour G, Saleh AL. Development and validation of an RP-HPLC method for the simultaneous determination of trimethoprim, sulfadimidine sodium and tylosin tartrate in an injectable solution formulation. J Appl Pharm Sci 2015;5:94-8.
15. Abbas R, Qorban BN, Abbas K. Simultaneous analysis of trimethoprim and sulphamethoxazole drug combinations in dosage forms by high-performance liquid chromatography. Iran Biomed J 2000;4:75-8.
16. Gurumurthy T, Monika M, Ashwini V. Development and validation RP-HPLC method for simultaneous estimation of sulfamethoxazole and trimethoprim. Indian J Res Pharm Biotechnol 2017;5:235-8.
17. Bonazzi D, Anddrisano V, Di Pietra AM, Cavrini V. Analysis of trimethoprim--sulfonamide drug combinations in dosage forms by UV spectroscopy and liquid chromatography (HPLC). Farmaco 1994;49:381-6.
18. Nachilobe P, Cassidy RM, Fesser ACE. Determination of trimethoprim in bovine serum high-performance liquid chromatography with confirmation by thermospray liquid chromathography-mass spectrometry. J Chromathogr 1993;616:243-52.
19. Gentleman MS, Burt HM, Kitts DD, McErlane KM. High-performance liquid chromatographic determination of sulphadiazine and trimethoprim in chinook salmon muscle tissue. J Chromatogr A 1993;633:105–10.
20. Batzias GC, Botsoglou NA, Kotsaki-Kovatsi VP, Kounenis G. New simple liquid chromatographic method for the determination of trimethoprim, sulfadiazine and n4-acetylsulfadiazine in plasma of broilers. J Chromatogr B Anal Technol Biomed Life Sci 2002;769:253-9.
21. Mashhour MG, Saleh AA. Development and validation of a stability-indicating HPLC method for the simultaneous determination of sulfadiazine sodium and trimethoprim in injectable solution formulation. Sci Pharm 2013;81:167-82.
22. Baere SD, Baert K, Croubels S, Busser JD, Wasch K, Backer PD. Determination and quantification of sulfadiazine and trimethoprim in swine tissues using liquid chromatography with ultraviolet and mass spectrometric detection. Analyst 2000;125:409-15.
23. Croubels S, Pascal W, Backer PD. Simultaneous determination of sulfadiazine and trimethoprim in animal feed by liquid chromatography with UV and tandem mass spectrometric detection. Anal Chim Acta 2002;473:183-94.
24. Croubels S, Baere SD, Backer PD. Comparison of a liquid chromatographic method with ultraviolet and ion-trap tandem mass spectrometric detection for the simultaneous determination of sulfadiazine and trimethoprim in plasma from dogs. J Chromatogr B 2003;788:167-78.
25. Sorensen LK, Elbaek TH. Simultaneous determination of trimethoprim, sulfadiazine, florfenicol and oxolinic acid in surface water by liquid chromatography-tandem mass spectrometry. Chromatographia 2004;60:287-91.
26. International Conference of Harmonization. Validation of Analytical Procedures: Text and Methodology. Geneva, Switzerland: ICH Q2 (R1); 2005.
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How to Cite
Pokala, R. V., K. Kumari, and H. B. Bollikola. “UV SPECTROPHOTOMETRY METHOD DEVELOPMENT AND VALIDATION OF SULFADIAZINE AND TRIMETHOPRIM IN COMBINED DOSAGE FORM”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 1, Jan. 2018, pp. 103-7, doi:10.22159/ijpps.2018v10i1.21767.
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