Anti-tuberculosis site-specific oral delivery system that enhances rifampicin bioavailability in a fixed-dose combination with isoniazid


The in vivo release segregation of rifampicin (RIF) and isoniazid (INH) has been proposed as a strategy to avoid RIF acid degradation, which is known as one of the main factors for reduced RIF bioavailability and can result in drug-resistant tuberculosis. So far, this strategy has been scarcely explored. The aims of this study were to investigate the stability and bioavailability of RIF after combination of a very fast release matrix of RIF with a sustained delivery system of INH. A series of INH-alginic acid complexes (AA-INH) was obtained and characterized. Independent and sequential release profile of AA-INH at biorrelevant media of pH 1.20 and 6.80 was explored. In addition, AA-INH was combined with a RIF-carboxymethylcellulose very fast release complex (CMC-RIF) obtained previously and subjected to acid dissolution assays to evaluate RIF acid stability and determine RIF and INH dissolution efficiencies. Finally, a pharmacokinetic study in dogs was carried out. The AA-INH was easily obtained in solid-state. Their characterization revealed its ionic nature, with a loading capacity of around 30%. The dissolution efficiencies (15 min) confirmed release segregation in acid media with 7.8 and 65.6% for AA-INH and CMC-RIF, respectively. INH release rate from the AA-INH system was slow in acid media and increased in simulated intestinal media. The complete release of INH was achieved after 2 h in simulated intestinal media in the sequential release experiments. The acid degradation of RIF was significantly reduced (36.7%) when both systems were combined and oral administration to dogs revealed a 42% increase in RIF bioavailability. In conclusion, CMC-RIF and AA-INH may be useful for the formulation of a site-specific solid dosage form to overcome some of the main obstacles in tuberculosis treatment.

Graphical abstract

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Data availability

All data generated or analyzed during this study are included in this published article (and its supplementary information file).


  1. 1.

    World Health Organisation. Global tuberculosis report. Geneva, Switzerland: Licence CC BY- NC-SA 3.0 IGO; 2018.

  2. 2.

    Singh S, Mariappan TT, Shankar R, Sarda N, Singh B. A critical review of the probable reasons for the poor/variable bioavailability of rifampicin from anti-tubercular fixed-dose combination (FDC) products, and the likely solutions to the problem. Int J Pharm. 2001;228:5–17.

    CAS  Article  Google Scholar 

  3. 3.

    World Health Organization. WHO guidelines on tuberculosis infection prevention and control 2019 update. Geneva, Switzerland: CC BY-NC-SA 3.0 IGO; 2019.

  4. 4.

    World Health Organization. Guidelines for treatment of drug-susceptible tuberculosis and patient care, 2017 update. Geneva, Switzerland: Licence: CC BY-NC-SA 3.0 IGO; 2017.

  5. 5.

    Blomberg B, Spinaci S, Fourie B, Laing R. The rationale for recommending fixed-dose combination tablets for treatment of tuberculosis. Bull World Health Organ. 2001;79:61–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Ellard GA, Fourie PB. Rifampicin bioavailability: a review of its pharmacology and the chemotherapeutic necessity for ensuring optimal absorption. Int J Tuberc Lung Dis. 1999;3:S301–8 discussion S317–21. [Accessed 2019 Aug 12]. Available from:

  7. 7.

    Mariappan TT, Singh S. Regional gastrointestinal permeability of rifampicin and isoniazid (alone and their combination) in the rat. Int J Tuberc Lung Dis. 2003;7:797–803.

    CAS  Google Scholar 

  8. 8.

    Singh S, Mariappan TT, Sharda N, Kumar S, Chakraborti AK. The reason for an increase in decomposition of rifampicin in the presence of isoniazid under acid conditions. Pharm Pharmacol Commun. 2000;6:405–10.

    CAS  Article  Google Scholar 

  9. 9.

    Avachat AM, Bhise SB. Tailored release drug delivery system for rifampicin and isoniazid for enhanced bioavailability of rifampicin. Pharm Dev Technol. 2011;16:127–36 Available from:

  10. 10.

    Prasad B, Singh S. LC-MS/TOF and UHPLC-MS/MS study of in vivo fate of rifamycin isonicotinyl hydrazone formed on oral co-administration of rifampicin and isoniazid. J Pharm Biomed Anal. Elsevier B.V.; 2010;52:377–83. [Accessed 2019 Aug 12]. Available from:

  11. 11.

    Singh S, Bhutani H, Mariappan TT. Quality problems of anti-tuberculosis fixed-dose combinations (Fdcs ): a way forward. Indian J Tuberc. 2006;53:201–5.

    Google Scholar 

  12. 12.

    Blomberg B, Fourie B. Fixed-dose combination drugs for tuberculosis: application in standardized treatment regimens. Drugs. 2003;63:535–53.

    CAS  Article  Google Scholar 

  13. 13.

    Pillai G, Fourie PB, Padayatchi N, Onyebujoh PC, McIlleron H, Smith PJ, et al. Recent bioequivalence studies on fixed-dose combination anti-tuberculosis drug formulations available on the global market. Int J Tuberc Lung Dis. 1999;3:S309–16 discussion S317-S321. [Accessed 2019 Aug 12]. Available from:

  14. 14.

    Panchagnula R, Agrawal S. Biopharmaceutic and pharmacokinetic aspects of variable bioavailability of rifampicin. Int J Pharm. 2004;271:1–4.

    CAS  Article  Google Scholar 

  15. 15.

    Zhu H, Guo SC, Hao LH, Liu CC, Wang B, Fu L, et al. Relative bioavailability of rifampicin in four Chinese fixed-dose combinations compared with rifampicin in free combinations. Chin Med J. 2015;128:433–7.

    CAS  Article  Google Scholar 

  16. 16.

    Luciani-Giacobbe LC, Ramírez-Rigo M V., Garro-Linck Y, Monti GA, Manzo RH, Olivera ME. Very fast dissolving acid carboxymethylcellulose-rifampicin matrix: Development and solid-state characterization. Eur J Pharm Sci. Elsevier B.V.; 2017;96:398–410. Available from:

  17. 17.

    Sosnik A, Carcaboso ÁM, Glisoni RJ, Moretton M A, Chiappetta D A. New old challenges in tuberculosis: potentially effective nanotechnologies in drug delivery. Adv Drug Deliv Rev. Elsevier B.V.; 2010;62:547–59. Available from:

  18. 18.

    Sosnik A, Seremeta KP, Imperiale JC, Chiappetta DA. Novel formulation and drug delivery strategies for the treatment of pediatric poverty-related diseases. Expert Opin Drug Deliv. 2012;9:303–23.

    CAS  Article  Google Scholar 

  19. 19.

    Bermúdez JM, Jimenez-Kairuz AF, Olivera ME, Allemandi DA, Manzo RH. A ciprofloxacin extended release tablet based on swellable drug polyelectrolyte matrices. AAPS PharmSciTech. 2008;9:924–30.

    Article  Google Scholar 

  20. 20.

    Ramírez Rigo MV, Allemandi DA, Manzo RH. Swellable drug-polyelectrolyte matrices (SDPM) of alginic acid. Characterization and delivery properties. Int J Pharm. 2006;322:36–43.

    Article  Google Scholar 

  21. 21.

    Jimenez-Kairuz AF, Llabot JM, Allemandi DA, Manzo RH. Swellable drug-polyelectrolyte matrices (SDPM) characterization and delivery properties. Int J Pharm. 2005;288:87–99 [Accessed 2019 Aug 2] Available from:

  22. 22.

    Ramírez Rigo M, Allemandi D, Manzo R. Swellable drug-polyelectrolyte matrices of drug-carboxymethylcellulose complexes. Characterization and delivery properties. Drug Deliv. 2009;16:108–15.

    Article  Google Scholar 

  23. 23.

    United States Pharmacopeial Convention. The United States Pharmacopeia, The National Formulary [USP 42 NF 37]. Rockville: United States Pharmacopoeial Convention; 2019.

    Google Scholar 

  24. 24.

    Singh H, Bhandari R, Kaur IP. Encapsulation of rifampicin in a solid lipid nanoparticulate system to limit its degradation and interaction with isoniazid at acidic pH. Int J Pharm. Elsevier B.V.; 2013;446:106–11. Available from:

  25. 25.

    WHO Expert Committee on Specifications for Pharmaceutical Preparations. Annex 11 Guidance on the selection of comparator pharmaceutical products for equivalence assessment of interchangeable multisource (generic) products. WHO Tech Rep Ser. 2002;161–80.

  26. 26.

    Sankar R, Sharda N, Singh S. Behavior of decomposition of rifampicin in the presence of isoniazid in the pH range 1–3. Drug Dev Ind Pharm. 2003;29:733–9.

    CAS  Article  Google Scholar 

  27. 27.

    Moretton MA, Hocht C, Taira C, Sosnik A. Rifampicin-loaded “flower-like” polymeric micelles for enhanced oral bioavailability in an extemporaneous liquid fixed-dose combination with isoniazid. Nanomedicine. 2014;9:1635–50.

    CAS  Article  Google Scholar 

  28. 28.

    Guzman ML, Romañuk CB, Sanchez MF, Giacobbe LCL. Urinary excretion of ciprofloxacin after administration of extended release tablets in healthy volunteers . Swellable drug-polyelectrolyte matrix versus bilayer tablets. Drug Deliv Transl Res. 2017.

  29. 29.

    Luciani-Giacobbe LC, Guzman ML, Manzo RH, Olivera ME. Validation of a simple isocratic HPLC-UV method for rifampicin and isoniazid quantification in human plasma. J Appl Pharm Sci. 2018;8:93–9 [Accessed 2019 Aug 2] Available from:

  30. 30.

    García MC, Guzman ML, Himelfarb MA, Litterio NJ, Olivera ME, Jimenez-Kairuz A. Preclinical pharmacokinetics of benznidazole-loaded interpolyelectrolyte complex-based delivery systems. Eur J Pharm Sci. Elsevier; 2018;122:281–91. Available from:

  31. 31.

    Borba A, Gómez-Zavaglia A, Fausto R. Molecular structure, infrared spectra, and photochemistry of isoniazid under cryogenic conditions. J Phys Chem A. 2009;113:9220–30.

    CAS  Article  Google Scholar 

  32. 32.

    Rastogi R, Sultana Y, Aqil M, Ali A, Kumar S, Chuttani K, et al. Alginate microspheres of isoniazid for oral sustained drug delivery. Int J Pharm. 2007;334:71–7.

    CAS  Article  Google Scholar 

  33. 33.

    Lavor EP, Freire FD, Aragão CFS, Raffin FN, De Lima E Moura TFA. Application of thermal analysis to the study of anti-tuberculosis drug compatibility. Part 1. J Therm Anal Calorim. 2012;108:207–12.

    CAS  Article  Google Scholar 

  34. 34.

    Alves R. Estudo termoanalítico e de compatibilidade fármaco-excipiente de rifampicina e alguns medicamentos utilizados na terapêutica da tuberculose. Universidade de Sao Paulo; 2007.

  35. 35.

    Soares JP, Santos JE, Chierice GO, Cavalheiro ETG. Thermal behavior of alginic acid and its sodium salt. Ecletica Quim. 2004;29:57–63.

    CAS  Article  Google Scholar 

  36. 36.

    Guzmán ML, Manzo RH, Olivera ME. Eudragit E100 as a drug carrier: the remarkable affinity of phosphate ester for dimethylamine. Mol Pharm. 2012;9:2424–33.

    Article  Google Scholar 

  37. 37.

    Ramírez Rigo M, Allemandi D, Manzo R. A linear free energy relationship treatment of the affinity between carboxymethylcellulose and basic drugs. Mol Pharm. 2004;1:383–6.

    Article  Google Scholar 

  38. 38.

    Quinteros DA, Rigo VR, Kairuz AFJ, Olivera ME, Manzo RH, Allemandi DA. Interaction between a cationic polymethacrylate (Eudragit E100) and anionic drugs. Eur J Pharm Sci. 2008;33:72–9.

    CAS  Article  Google Scholar 

  39. 39.

    Olivera ME, Manzo RH, Alovero F, Alvaro F. Polyelectrolyte-drug ionic complexes as nanostructured drug carriers to design solid and liquid oral delivery systems. Nanostruct Oral Med. Amsterdam, Oxford, Cambridge: Elsevier. 2017:365–408.

  40. 40.

    Liew CV, Chan LW, Ching AL, Heng PWS. Evaluation of sodium alginate as drug release modifier in matrix tablets. Int J Pharm. 2006;309:25–37.

    CAS  Article  Google Scholar 

  41. 41.

    Gohel MC, Sarvaiya KG. A novel dosage form of rifampicin and isoniazid with improved functionality. AAPS PharmSciTech. 2007;44:22–7.

    Google Scholar 

  42. 42.

    Wang Y, Liu H, Liu K, Sun J, He Z. Design and evaluation of enteric-coated tablets for rifampicin and isoniazid combinations. Pharm Dev Technol. 2013;18:401–6 [Accessed 2019 Aug 2] Available from:

  43. 43.

    Genina N, Boetker JP, Colombo S, Harmankaya N, Rantanen J, Bohr A. Anti-tuberculosis drug combination for controlled oral delivery using 3D printed compartmental dosage forms: from drug product design to in vivo testing. J Control Release. Elsevier; 2017;268:40–8. Available from:

  44. 44.

    Panacea Biotec. Xeed. 2000 [cited 2016 Jan 2]. [Accessed 2019 Aug 2] Available from:

  45. 45.

    Clyna S.A. P.R. vademecum [Internet]. 2019 [Accessed 2019 Aug 2]. Available from:

  46. 46.

    U.S. Food and Drug Administration. Drugs@FDA: FDA Approved Drug Products [Internet]. Search Results for “rifampin.” 2019 [Accessed 2019 Aug 2]. p. 1. Available from:

  47. 47.

    Long MW, Snider DEJ, Farer LS. U.S. Public Health Service Cooperative trial of three rifampin-isoniazid regimens in treatment of pulmonary tuberculosis. Am Rev Respir Dis. 1979;119:879–94. [Accessed 2019 Aug 2] Available from:

  48. 48.

    Dressman JB. Comparison of canine and human gastrointestinal physiology. Pharm Res. 1986;3:123–31.

    CAS  Article  Google Scholar 

  49. 49.

    World Health Organization. Global tuberculosis report 2014. Geneva: WHO; 2014. [Accessed 2019 Aug 2] Available from:

Download references


Luciani-Giacobbe thanks CONICET for her postdoctoral scholarship. We thank Dr. Paul Hobson, native speaker, for revision of the manuscript.


This work was supported by Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, grant number 11220120100461), the Fondo para la Investigación Científica y Tecnológica (FonCyT, grant number PICT 0173), and the Universidad Nacional de Córdoba (SECYT-UNC, grant number 162/12).

Author information



Corresponding author

Correspondence to María Eugenia Olivera.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal rights and informed consent

All institutional and national guidelines for the care and use of laboratory animals were followed.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(PPTX 207 kb).

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Luciani-Giacobbe, L.C., Lorenzutti, A.M., Litterio, N.J. et al. Anti-tuberculosis site-specific oral delivery system that enhances rifampicin bioavailability in a fixed-dose combination with isoniazid. Drug Deliv. and Transl. Res. 11, 894–908 (2021).

Download citation


  • Swellable drug-polyelectrolyte matrices
  • Solid state characterization
  • Dissolution
  • Stability
  • Pharmacokinetics