AAPS PharmSciTech

, Volume 14, Issue 1, pp 425–434 | Cite as

Development and Validation of a Discriminative Dissolution Test for Betamethasone Sodium Phosphate and Betamethasone Dipropionate Intramuscular Injectable Suspension

  • Alice Simon
  • Vinícius Raphael de Almeida Borges
  • Lucio Mendes Cabral
  • Valéria Pereira de Sousa
Research Article


The intramuscular administration of the injectable suspension betamethasone sodium phosphate (BSP) and betamethasone dipropionate (BD) has immediate therapeutic activity due to solubilized BSP and prolonged activity resulting from the slow release of BD micro-crystals. The purpose of this study was to develop and validate a dissolution method for BD in intramuscular injectable suspensions with detection by high-performance liquid chromatography (HPLC) method. Five commercial products presented a distribution of particle sizes, ranging between 7.43 and 40.25 μm as measured by laser diffraction. It was also found that particle sizes differed between batches of the same product. The different products were tested using the paddle apparatus, with stirring speeds of 25 and 50 rpm in 300 mL of phosphate buffer; simulated body fluid, muscle fluid, and synovial fluid were used as biorelevant dissolution media at 37 ± 0.5°C. It was verified that not only does average particle size affect the dissolution rate, but also the mode and the polydispersity index of the particles. Discriminatory power was obtained using the in vitro dissolution method with 0.1 M sodium phosphate buffer pH 7.4 containing 0.1% sodium lauryl sulfate and a stirring speed of 50 rpm. The HPLC-method is linear, precise, selective, and accurate for the quantification of BSP and BD in dissolution profile testing. This dissolution method can be utilized as a method to control the quality of these injectable suspensions.

Key words

dipropionate betamethasone dissolution test intramuscular injectable suspensions simulated muscular fluid sodium phosphate betamethasone 



This work was supported by FAPERJ, Edital CAPES Nanobiotecnologia 2008, and CNPq. We are grateful to Michelle Parvatiyar for her English review.


  1. 1.
    Diprospan® bull. Mantecorp Indústria Química e Farmacêutica LTDA, Rio de Janeiro; 2012.[27314-1-0].PDF. Accessed 10 Sept 2012.
  2. 2.
    Zou J-J, Dai L, Ding L, Xiao D-W, Bin Z-Y, Fan H-W, Liu L, Wang G-J. Determination of betamethasone and betamethasone 17-monopropionate in human plasma by liquid chromatography–positive/negative electrospray ionization tandem mass spectrometry. J Chromatogr B. 2008;873:159–64.CrossRefGoogle Scholar
  3. 3.
    Gazelka HM, Burgher AH, Huntoon MA, Mantilla CB, Hoelzer BC. Determination of the particulate size and aggregation of clonidine and corticosteroids for epidural steroid injection. Pain Physician. 2012;15:87–93.PubMedGoogle Scholar
  4. 4.
    Rasenack N, Hartenhauer H, Müller BW. Microcrystals for dissolution rate enhancement of poorly water-soluble drugs. Int J Pharm. 2003;254:137–45. doi: 10.1016/S0378-5173(03)00005-X.CrossRefPubMedGoogle Scholar
  5. 5.
    Pillay V, Fassihi R. Unconventional dissolution methodologies. J Pharm Sci. 1999;88:843–51. doi: 10.1021/js990139b.CrossRefPubMedGoogle Scholar
  6. 6.
    Yuksel N, Kanik AE, Baykara T. Comparison of in vitro dissolution profiles by ANOVA-based, model-dependent and -independent methods. Int J Pharm. 2000;209:57–67. doi: 10.1016/S0378-5173(00)00554-8.CrossRefPubMedGoogle Scholar
  7. 7.
    Dressman JB, Amidon GL, Reppas C, Shah VP. Dissolution testing as a prognostic tool for oral drug absorption: immediate release dosage forms. Pharm Res. 1998;15:11–22. doi: 10.1023/A:1011984216775.CrossRefPubMedGoogle Scholar
  8. 8.
    Löbenberg R, Krämer J, Shah VP. Dissolution testing as a prognostic tool for oral drug absortion: dissolution behavior of glibenclamide. Pharm Res. 2000;17:439–44. doi: 10.1023/A:1007529020774.CrossRefPubMedGoogle Scholar
  9. 9.
    The United States Pharmacopoeia–National Formulary. <711 > Dissolution. 34th ed, Rockville, MD: USP Convention Inc.; 2011Google Scholar
  10. 10.
    Siewert M, Dressman J, Brown CK, Shah VP, Aiache JM, Aoyagi N, et al. FIP/AAPS Guidelines to dissolution in vitro release testing of novel/special dosage forms. AAPS PharmSciTech. 2003;4(1):E7. doi: 10.1208/pt040107.CrossRefPubMedGoogle Scholar
  11. 11.
    Azarmi S, Roa W, Löbenberg R. Current perspectives in dissolution testing of conventional and novel dosage forms. Int J Pharm. 2007;328:12–21. doi: 10.1016/j.ijpharm.2006.10.001.CrossRefPubMedGoogle Scholar
  12. 12.
    Fonseca LB, Labastie M, Sousa VP, Volpato NM. Development and validation of a discriminative dissolution test for nimesulide suspension. AAPS PharmSciTech. 2009;10:1145–52. doi: 10.1208/s12249-009-9320-4.CrossRefPubMedGoogle Scholar
  13. 13.
    Bisrat M, Nyström C. Physicochemical aspects of drug release. VIII. The relation between particle size and surface specific dissolution rate in agitated suspensions. Int J Pharm. 1988;47:223–31. doi: 10.1016/0378-5173(88)90235-9.CrossRefGoogle Scholar
  14. 14.
    Simões S, Sousa A, Figueiredo M. Dissolution rate studies of pharmaceutical multisized powders—a practical approach using the Coulter method. Int J. 1996;127:283–91. doi: 10.1016/0378-5173(95)04312-8.Google Scholar
  15. 15.
    Burgess DJ, Hussain AS, Ingallinera TS, Chen M. Assuring quality and performance of sustained and controlled release parenteral. AAPS Pharm Sci. 2002;4:134–8. doi: 10.1208/ps040205.CrossRefGoogle Scholar
  16. 16.
    Alam A, Ahuja A, Baboota S, Gidwani SK, Ali J. Formulation and evaluation of pharmaceutically equivalent parenteral depot suspension of methyl prednisolone acetate. Indian J Pharm Sci. 2009;71:30–4. doi: 10.4103/0250-474X.CrossRefPubMedGoogle Scholar
  17. 17.
    The United States Pharmacopoeia–National Formulary 29. <1092 > The dissolution procedure: development and validation. 34th ed, Rockville, MD: USP Convention Inc.; 2011.Google Scholar
  18. 18.
    Simon A, Cabral LM, Sousa VP. Desenvolvimento e validação de método analítico por CLAE para a quantificação simultânea de dipropionato de betametasona e fosfato sódico de betametasona em suspensão injetável. Quim Nova. 2012;35:593–600.CrossRefGoogle Scholar
  19. 19.
    ICH, validation of analytical procedures: text and methodology (Q2R1), in International Conference on Harmonization; November 2005.Google Scholar
  20. 20.
    Shabir GA. Validation of high-performance liquid chromatography methods for pharmaceutical analysis: understanding the differences and similarities between validation requirements of the US Food and Drug Administration, the US Pharmacopeia and the International Conference on Harmonization. J Chromatogr A. 2003;987:57–66. doi: 10.1016/S0021-9673(02)01536-4.CrossRefPubMedGoogle Scholar
  21. 21.
    Rossi RC, Dias CL, Bajerski L, Bergold AM, Fröehlich PE. Development and validation of discriminating method of dissolution for fosamprenavir tablets based on in vivo data. J Pharm Biomed Anal. 2011;54:439–44. doi: 10.1016/j.jpba.2010.09.004.CrossRefPubMedGoogle Scholar
  22. 22.
    Lindenberg M, Wiegand C, Dressman JB. Comparison of the adsorption of several drugs to typical filter materials. Dissolution Technol. 2005;12:22–5.Google Scholar
  23. 23.
    Sarkar A, Rano R, Mishra KK, Sinha IN. Particle size distribution profile of some Indian fly ash—a comparative study to assess their possible uses. Fuel Process Technol. 2005;86:1221–38. doi: 10.1016/j.fuproc.2004.12.002.CrossRefGoogle Scholar
  24. 24.
    Ghazal HS, Dyas AM, Ford JL, Hutcheon GA. In vitro evaluation of the dissolution behavior of itraconazole in bio-relevant media. Int J Pharm. 2009;366:117–23. doi: 10.1016/j.ijpharm.2008.09.003.CrossRefPubMedGoogle Scholar
  25. 25.
    Marques MRC, Loebenberg R, Almukainzi M. Simulated biological fluids with possible application in dissolution testing. Dissolution Technol. 2011;18:15–28.Google Scholar
  26. 26.
    Fogh-Andersen N, Altura BM, Altura BT, Siggaard-Andersen O. Composition of interstitial fluid. Clin Chem. 1995;41:1522–5.PubMedGoogle Scholar
  27. 27.
    Sorenson MM, Silva AC, Gouveia CS, Sousa VP, Oshima W, Ferro JA, et al. Concerted action of the high affinity calcium binding sites in skeletal muscle troponin C. J Biol Chem. 1995;270:9770–7.CrossRefPubMedGoogle Scholar
  28. 28.
    Sousa VP, Pinto JRDO, Sorenson MM. Ionic interventions that alter the association of troponin c c-domain with the thin filaments of vertebrate striated muscle. Biochim Biophys Acta. 2006;1760:272–82. doi: 10.1016/j.bbagen.2005.09.014.CrossRefPubMedGoogle Scholar
  29. 29.
    FDA. Guidance for industry: dissolution testing of immediate release solid oral dosage forms. U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER); August 1997.Google Scholar
  30. 30.
    Dokoumetzidis A, Macheras P. A century of dissolution research: from Noyes and Whitney to the Biopharmaceutics Classification System. Int J Pharm. 2006;321:1–11. doi: 10.1016/j.ijpharm.2006.07.011.CrossRefPubMedGoogle Scholar
  31. 31.
    Shen J, Burgess DJ. Accelerated in-vitro release testing methods for extended-release parenteral dosage forms. J Pharm Pharmacol. 2012;64:986–96.CrossRefPubMedGoogle Scholar
  32. 32.
    Larsen C, Ostergaard J, Larsen SW, Jensen H, Jacobsen S, Lindegaard C, et al. Intra-articular depot formulation principles: role in the management of postoperative pain and arthritic disorders. J Pharm Sci. 2008;97:4622–54.CrossRefPubMedGoogle Scholar
  33. 33.
    R. M. Patel. Parenteral suspension: an overview. Int J Curr Pharm Res. 2010;2,3:3–13.Google Scholar
  34. 34.
    The United States Pharmacopoeia, National Formulary 29. 34th ed., Supp. 1. Rockville, MD: U.S. Pharmacopoeial Convention; 2011.Google Scholar
  35. 35.
    Conzone SD, Brown RF, Day DE, Ehrhardt GJ. In vitro and in vivo dissolution behavior of a dysprosium lithium borate glass designed for the radiation synovectomy treatment of rheumatoid arthritis. J Biomed Mater Res. 2002;60:260–8. doi: 10.1002/jbm.10047.CrossRefPubMedGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2013

Authors and Affiliations

  • Alice Simon
    • 1
  • Vinícius Raphael de Almeida Borges
    • 1
  • Lucio Mendes Cabral
    • 1
  • Valéria Pereira de Sousa
    • 1
  1. 1.Department of Pharmaceutics, Faculty of PharmacyFederal University of Rio de JaneiroRio de JaneiroBrazil

Personalised recommendations