Statistical analysis of solid lipid nanoparticles produced by high-pressure homogenization: a practical prediction approach

  • Matilde Durán-Lobato
  • Alicia Enguix-González
  • Mercedes Fernández-Arévalo
  • Lucía Martín-Banderas
Research Paper


Lipid nanoparticles (LNPs) are a promising carrier for all administration routes due to their safety, small size, and high loading of lipophilic compounds. Among the LNP production techniques, the easy scale-up, lack of organic solvents, and short production times of the high-pressure homogenization technique (HPH) make this method stand out. In this study, a statistical analysis was applied to the production of LNP by HPH. Spherical LNPs with mean size ranging from 65 nm to 11.623 μm, negative zeta potential under –30 mV, and smooth surface were produced. Manageable equations based on commonly used parameters in the pharmaceutical field were obtained. The lipid to emulsifier ratio (R L/S) was proved to statistically explain the influence of oil phase and surfactant concentration on final nanoparticles size. Besides, the homogenization pressure was found to ultimately determine LNP size for a given R L/S, while the number of passes applied mainly determined polydispersion. α-Tocopherol was used as a model drug to illustrate release properties of LNP as a function of particle size, which was optimized by the regression models. This study is intended as a first step to optimize production conditions prior to LNP production at both laboratory and industrial scale from an eminently practical approach, based on parameters extensively used in formulation.


Solid lipid nanoparticles High-pressure homogenization Statistical analysis Regression model Particle size prediction Mathematical model Drug release 



M. D. L. thanks University of Seville for a Grant from IV Research Plan of University of Seville. L. M. B. is especially grateful to Junta de Andalucía (Spain) for financial support (Project No. P09-CTS5029). Microscopy Services (Centro de Investigación, Tecnología e Innovación de la Universidad de Sevilla, CITIUS) technical support is also grateful. Authors also thank Dr. Álvarez-Fuentes for technical support.


  1. Araujo J, Gonzalez-Mira E, Egea MA, Garcia ML, Souto EB (2010) Optimization and physicochemical characterization of a triamcinolone acetonide-loaded NLC for ocular antiangiogenic applications. Int J Pharm 393:167–175CrossRefGoogle Scholar
  2. Biasutti M, Venir E, Marchesini G, Innocente N (2010) Rheological properties of model dairy emulsions as affected by high pressure homogenization. Innov Food Sci Emerg Technol 11:580–586CrossRefGoogle Scholar
  3. Bondì ML, Craparo EF, Giammona G, Drago F (2010) Brain-targeted solid lipid nanoparticles containing riluzole: preparation, characterization and biodistribution. Nanomedicine (Lond, Engl) 5:25–32CrossRefGoogle Scholar
  4. Charcosset C, El-Harati A, Fessi H (2005) Preparation of solid lipid nanoparticles using a membrane contactor. J Control Release 108:112–120CrossRefGoogle Scholar
  5. Chattopadhyay P, Shekunov BY, Yim D, Cipolla D, Boyd B, Farr S (2007) Production of solid lipid nanoparticle suspensions using supercritical fluid extraction of emulsions (SFEE) for pulmonary delivery using the AERx system. Adv Drug Deliv Rev 59:444–453CrossRefGoogle Scholar
  6. Das S, Chaudhury A (2011) Recent advances in lipid nanoparticle formulations with solid matrix for oral drug delivery. AAPS PharmSciTech 12:62–76CrossRefGoogle Scholar
  7. El-Harati AA, Charcosset C, Fessi H (2006) Influence of the formulation for solid lipid nanoparticles prepared with a membrane contactor. Pharm Dev Technol 11:153–157CrossRefGoogle Scholar
  8. Floury J, Desrumaux A, Lardières J (2000) Effect of high-pressure homogenization on droplet size distributions and rheological properties of model oil-in-water emulsions. Innov Food Sci Emerg Technol 1:127–134CrossRefGoogle Scholar
  9. García-Fuentes M, Torres D, Alonso MJ (2002) Design of lipid nanoparticles for the oral delivery of hydrophilic macromolecules. Colloid Surf B 27:159–168CrossRefGoogle Scholar
  10. Gasco MR (1997) Solid lipid nanospheres from warm microemulsion. Pharm Technol Eur 9:32–42Google Scholar
  11. Håkansson A, Fuchs L, Innings F, Revstedt J, Trägårdh C (2011) On flow-fields in a high pressure homogenizer and its implication on drop fragmentation. Procedia Food Sci 1:1353–1358CrossRefGoogle Scholar
  12. Heurtault B, Saulnier P, Pech B, Proust JE, Benoit JP (2002) A novel phase inversion-based process for the preparation of lipid nanocarriers. Pharm Res 19:875–880CrossRefGoogle Scholar
  13. Hu FQ, Yuan H, Zhang HH, Fang M (2002) Preparation of solid lipid nanoparticles with clobetasol propionate by a novel solvent diffusion method in aqueous system and physicochemical characterization. Int J Pharm 239:121–128CrossRefGoogle Scholar
  14. Innocente N, Biasutti M, Venir E, Spaziani M, Marchesini E (2009) Effect of high-pressure homogenization on droplet size distribution and rheological properties of ice cream mixes. J Dairy Sci 92:1864–1875CrossRefGoogle Scholar
  15. Jafari S, Assadpoor E, He Y, Bhandari B (2008) Re-coalescence of emulsion droplets during high-energy emulsification. Food Hydrocoll 22:1191–1202CrossRefGoogle Scholar
  16. Kluge J, Muhrer G, Mazzotti M (2012) High pressure homogenization of pharmaceutical solids. J Supercrit Fluids 66:380–388CrossRefGoogle Scholar
  17. Liedtke S, Wissing S, Müller RH, Mäder K (2000) Influence of high pressure homogenisation equipment on nanodispersions characteristics. Int J Pharm 196:183–185CrossRefGoogle Scholar
  18. Liu J, Hu W, Chen H, Ni Q, Xu H, Yang X (2007) Isotretinoin-loaded solid lipid nanoparticles with skin targeting for topical delivery. Int J Pharm 328:191–195CrossRefGoogle Scholar
  19. Maindarkar SN, Raikar NB, Bongers P, Henson MA (2012) Incorporating emulsion drop coalescence into population balance equation models of high pressure homogenization. Colloids Surf A 396:63–73CrossRefGoogle Scholar
  20. McClements DJ (2005) Food emulsions principles, practices, and techniques, 2nd edn. CRC Press, Boca RatonGoogle Scholar
  21. Mehnert W, Mäder K (2001) Solid lipid nanoparticles: production, characterization and applications. Adv Drug Deliv Rev 47:165–196CrossRefGoogle Scholar
  22. Mishra DK, Dhote V, Bhatnagar P, Mishra PK (2012) Engineering solid lipid nanoparticles for improved drug delivery: promises and challenges of translational research. Drug Deliv Transl Res 2:238–253CrossRefGoogle Scholar
  23. Mohr K-H (1987a) High-pressure homogenization. Part I. Liquid–liquid dispersion in turbulence fields of high energy density. J Food Eng 6:177–186CrossRefGoogle Scholar
  24. Mohr K-H (1987b) The influence of cavitation on liquid–liquid dispersion in turbulence fields of high energy density. J Food Eng 6:311–324CrossRefGoogle Scholar
  25. Mucho M, Maincent P, Müller RH (2008) Lipid nanoparticles with a solid matrix (LNP, NLC, LDC) for oral drug delivery. Drug Dev Ind Pharm 34:1394–1405CrossRefGoogle Scholar
  26. Müller RH, Schwarz C, Zur Mühlen A, Mehnert W (1994) Incorporation of lipophilic drugs and drug release profiles of solid lipid nanoparticles (LNP). Proc Int Symp Control Release Bioact Mater 21:146Google Scholar
  27. Müller RH, Maassen S, Schwarz C, Mehnert W (1997) Solid lipid nanoparticles (LNP) as potential carrier for human use: interaction with human granulocytes. J Control Release 47:261–269CrossRefGoogle Scholar
  28. Müller RH, Dingler A, Schneppe T, Gohla S (2000a) Large scale production of solid lipid nanoparticles (SLN) and nanosuspensions (DissoCubes). In: Wise D (ed) Handbook of pharmaceutical controlled release technology [e-book]. Marcel Dekker, New York, pp 359–376Google Scholar
  29. Müller RH, Mäder K, Gohla S (2000b) Solid lipid nanoparticles (LNP) for controlled drug delivery—a review of the state of the art. Eur J Pharm Biopharm 50:161–177CrossRefGoogle Scholar
  30. Pardeike J, Hommoss A, Müller RH (2009) Lipid nanoparticles (LNP, NLC) in cosmetic and pharmaceutical dermal products. Int J Pharm 366:170–184CrossRefGoogle Scholar
  31. Priano L, Esposti D, Castagna G, De Medici C, Fraschini F, Gasco MR, Mauro A (2007) Solid lipid nanoparticles incorporating melatonin as new model for sustained oral and transdermal delivery systems. J Nanosci Nanotechnol 7:3596–3601CrossRefGoogle Scholar
  32. Puglia C, Blasi P, Rizza L, Schoubben A, Bonina F, Rossi C, Ricci M (2008) Lipid nanoparticles for prolonged topical delivery: an in vitro and in vivo investigation. Int J Pharm 357:295–304CrossRefGoogle Scholar
  33. Qian C, McClements DJ (2010) Formation of nanoemulsions stabilized by model food-grade emulsifiers using high-pressure homogenization: factors affecting particle size. Food Hydrocoll 25:1000–1008CrossRefGoogle Scholar
  34. Saheki A, Seki J, Nakanishi T, Tamai I (2012) Effect of back pressure on emulsification of lipid nanodispersions in a high-pressure homogenizer. Int J Pharm 422:489–494CrossRefGoogle Scholar
  35. Salmaso S, Elvassore N, Bertucco A, Caliceti P (2009) Production of solid lipid submicron particles for protein delivery using a novel supercritical gas-assisted melting atomization process. J Pharm Sci 98:640–650CrossRefGoogle Scholar
  36. Schäfer-Korting M, Mehnert W, Korting HC (2007) Lipid nanoparticles for improved topical application of drugs for skin diseases. Adv Drug Deliv Rev 59:427–443CrossRefGoogle Scholar
  37. Schubert MA, Müller-Goymann CC (2003) Solvent injection as a new approach for manufacturing lipid nanoparticles—evaluation of the method and process parameters. Eur J Pharm Biopharm 55:125–131CrossRefGoogle Scholar
  38. Sebti T, Amighi K (2006) Preparation and in vitro evaluation of lipidic carriers and fillers for inhalation. Eur J Pharm Biopharm 63:51–58CrossRefGoogle Scholar
  39. Severino P, Santana MHA, Souto EB (2012) Optimizing LNP and NLC by 22 full factorial design: effect of homogenization technique. Mater Eng C 32:1375–1379CrossRefGoogle Scholar
  40. Sinha VR, Srivastava S, Goel H, Jindal V (2010) Solid lipid nanoparticles (SLN’S)—trends and implications in drug targeting. Int J Adv Pharm Sci 1:212–238Google Scholar
  41. Sjöstrom B, Bergenstahl B (1992) Preparation of submicron drug particles in lecithin-stabilized o/w emulsions: I. Model studies of the precipitation of cholesteryl acetate. Int J Pharm 84:107–116CrossRefGoogle Scholar
  42. Souto EB, Müller RH (2006) Investigation of the factors influencing the incorporation of clotrimazole in LNP and NLC prepared by hot high-pressure homogenization. J Microencapsul 23:377–388CrossRefGoogle Scholar
  43. Trombino S, Cassano R, Muzzalupo R, Pingitore A, Cione E, Picci N (2009) Stearyl ferulate-based solid lipid nanoparticles for the encapsulation and stabilization beta-carotene and alpha-tocopherol. Colloids Surf B 72:181–187CrossRefGoogle Scholar
  44. Trotta M, Debernardi F, Caputo O (2003) Preparation of solid lipid nanoparticles by a solvent emulsification–diffusion technique. Int J Pharm 257:153–160CrossRefGoogle Scholar
  45. Varshosaz J, Ghaffari S, Khoshayand MR, Atyabi F, Azarmi S, Kobarfard F (2010) Development and optimization of solid lipid nanoparticles of amikacin by central composite design. J Liposome Res 20:97–104CrossRefGoogle Scholar
  46. Vitorino C, Carvalho FA, Almeida AJ, Sousa JJ, Pais AACC (2011) The size of solid lipid nanoparticles: an interpretation from experimental design. Colloids Surf B 84:117–130CrossRefGoogle Scholar
  47. Walstra P (1993) Principles of emulsion formation. Chem Eng Sci 48:333–349CrossRefGoogle Scholar
  48. Wooster TJ, Golding M, Sanguansri P (2008) Impact of oil type on nanoemulsion formation and Ostwald ripening stability. Langmuir 24:12758–12765CrossRefGoogle Scholar
  49. Zur Mühlen A, Mehnert W (1998) Drug release and release mechanism of prednisolone loaded solid lipid nanoparticles. Pharmazie 53:552–555Google Scholar
  50. Zur Mühlen A, Schwarz C, Mehnert W (1998) Solid lipid nanoparticles (LNP) for controlled drug delivery–drug release and release mechanism. Eur J Pharm Biopharm 45:149–155CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Matilde Durán-Lobato
    • 1
  • Alicia Enguix-González
    • 2
  • Mercedes Fernández-Arévalo
    • 1
  • Lucía Martín-Banderas
    • 1
  1. 1.Dpto. Farmacia y Tecnología Farmacéutica, Facultad de FarmaciaUniversidad de SevillaSevilleEspaña
  2. 2.Dpto. Estadística e Investigación Operativa, Facultad de MatemáticasUniversidad de SevillaSevilleEspaña

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