AAPS PharmSciTech

, Volume 19, Issue 5, pp 2383–2394 | Cite as

Unstructured Formulation Data Analysis for the Optimization of Lipid Nanoparticle Drug Delivery Vehicles

  • Jessica Silva
  • Maria Mendes
  • Tânia Cova
  • João Sousa
  • Alberto Pais
  • Carla VitorinoEmail author
Research Article


Designing nanoparticle formulations with features tailored to their therapeutic targets in demanding timelines assumes increased importance. In this context, nanostructured lipid carriers (NLCs) offer an excellent example of a drug delivery nanosystem that has been broadly explored in the treatment of glioblastoma multiforme (GBM). Distinct fundamental NLC quality attributes can be harnessed to fit this purpose, namely particle size, size distribution, and zeta potential. These critical aspects intrinsically depend on the formulation components, influencing drug loading capacity, drug release, and stability of the NLCs. Wide variations in their composition, including the type of lipids and other surface modifier excipients, lead to differences on these parameters. NLC target product profile involves small mean particle sizes, narrow size distributions, and absolute values of zeta potential higher than 30 mV. In this work, a wealth of data previously obtained in experiments on NLC preparation, encompassing, e.g., results of preliminary studies and those of intermediate formulations, is analyzed in order to extract information useful in further optimization studies. Principal component analysis (PCA) and partial least squares (PLS) are performed to evaluate the influence of NLC composition on the respective characteristics. These methods provide a rapid and discriminatory analysis for establishing a preformulation framework, by selecting the most suitable types of lipids, surfactants, surface modifiers, and drugs, within the set of investigated variables. The results have direct implications in the optimization of formulation and processes.


glioblastoma multiforme (GBM) nanostructured lipid carriers (NLCs) critical quality attributes (CQAs) multivariate analysis 


Funding Information

The study was financially supported by FEDER Funds through the Operational Program Competitiveness Factors - COMPETE 2020 and by the Fundação para a Ciência e a Tecnologia (FCT), the Portuguese Agency for Scientific Research, through the projects no. 016648 (Ref. POCI-01-0145-FEDER-016648) and POCI-01-0145-FEDER-007440 - Center for Neurosciences and Cell Biology (CNC). The Coimbra Chemistry Centre is supported by the FCT through the projects PEst-OE/QUI/UI0313/2014 and POCI-01-0145-FEDER-007630. Maria Mendes and Tânia Cova were also supported, respectively, by the PhD research grants SFRH/BD/133996/2017 and SFRH/BD/95459/2013, assigned by FCT.

Supplementary material

12249_2018_1078_MOESM1_ESM.docx (31 kb)
ESM 1 (DOCX 31 kb)


  1. 1.
    Shah R, Eldridge D, Palombo E, Harding I. Lipid nanoparticles: production, characterization and stability. New York: Springer International Publishing; 2015.Google Scholar
  2. 2.
    Tapeinos C, Battaglini M, Ciofani G. Advances in the design of solid lipid nanoparticles and nanostructured lipid carriers for targeting brain diseases. J Control Release. 2017;264:306–32.CrossRefGoogle Scholar
  3. 3.
    Khan S, Baboota S, Ali J, Khan S, Narang RS, Narang JK. Nanostructured lipid carriers: an emerging platform for improving oral bioavailability of lipophilic drugs. Int J Pharm Investig. 2015;5(4):182–91.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Liu CH, Wu CT. Optimization of nanostructured lipid carriers for lutein delivery. Colloids Surf A Physicochem Eng Asp. 2010;353(2):149–56.CrossRefGoogle Scholar
  5. 5.
    Muchow M, Maincent P, Müller RH. Lipid nanoparticles with a solid matrix (SLN®, NLC®, LDC®) for oral drug delivery. Drug Dev Ind Pharm. 2008;34(12):1394–405.CrossRefGoogle Scholar
  6. 6.
    Song S, Mao G, Du J, Zhu X. Novel RGD containing, temozolomide-loading nanostructured lipid carriers for glioblastoma multiforme chemotherapy. Drug Delivery. 2016;23(4):1404–8.CrossRefGoogle Scholar
  7. 7.
    Qu J, Zhang L, Chen Z, Mao G, Gao Z, Lai X, et al. Nanostructured lipid carriers, solid lipid nanoparticles, and polymeric nanoparticles: which kind of drug delivery system is better for glioblastoma chemotherapy? Drug Delivery. 2016;23(9):3408–16.CrossRefGoogle Scholar
  8. 8.
    Karim R, Palazzo C, Evrard B, Piel G. Nanocarriers for the treatment of glioblastoma multiforme: current state-of-the-art. J Control Release. 2016;227:23–37.CrossRefGoogle Scholar
  9. 9.
    Pourgholi F, Farhad JN, Kafil HS, Yousefi M. Nanoparticles: novel vehicles in treatment of glioblastoma. Biomed Pharmacother. 2016;77:98–107.CrossRefGoogle Scholar
  10. 10.
    Jain K. Use of nanoparticles for drug delivery in glioblastoma multiforme. Expert Rev Neurother. 2007;7(4):363–72.CrossRefGoogle Scholar
  11. 11.
    Tzeng SY, Green JJ. Therapeutic nanomedicine for brain cancer. Ther Deliv. 2013;4(6):687–704.CrossRefGoogle Scholar
  12. 12.
    Iacob G, Dinca EB. Current data and strategy in glioblastoma multiforme. J Med Life. 2009;2(4):386–93.PubMedPubMedCentralGoogle Scholar
  13. 13.
    Urbańska K, Sokołowska J, Szmidt M, Sysa P. Glioblastoma multiforme - an overview. Contemp Oncol. 2014;18(5):307–12.Google Scholar
  14. 14.
    Ching J, Amiridis S, Stylli SS, Morokoff AP, O’Brien TJ, Kaye AH. A novel treatment strategy for glioblastoma multiforme and glioma associated seizures: increasing glutamate uptake with PPARγ agonists. J Clin Neurosci. 2015;22(1):21–8.CrossRefGoogle Scholar
  15. 15.
    Alifieris C, Trafalis DT. Glioblastoma multiforme: pathogenesis and treatment. Pharmacol Ther. 2015;152(Supplement C):63–82.CrossRefGoogle Scholar
  16. 16.
    Kim SS, Harford JB, Pirollo KF, Chang EH. Effective treatment of glioblastoma requires crossing the blood–brain barrier and targeting tumors including cancer stem cells: the promise of nanomedicine. Biochem Biophys Res Commun. 2015;468(3):485–9.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJB, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352(10):987–96.CrossRefGoogle Scholar
  18. 18.
    Lee SY. Temozolomide resistance in glioblastoma multiforme. Genes Dis. 2016;3(3):198–210.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Zhang H, Gao S. Temozolomide/PLGA microparticles and antitumor activity against glioma C6 cancer cells in vitro. Int J Pharm. 2007;329(1):122–8.CrossRefGoogle Scholar
  20. 20.
    Sordillo LA, Sordillo PP, Helson L. Curcumin for the treatment of glioblastoma. Anticancer Res. 2015;35(12):6373–8.Google Scholar
  21. 21.
    Zhuang W, Long L, Zheng B, Ji W, Yang N, Zhang Q, et al. Curcumin promotes differentiation of glioma-initiating cells by inducing autophagy. Cancer Sci. 2012;103(4):684–90.CrossRefGoogle Scholar
  22. 22.
    Luthra PM, Lal N. Prospective of curcumin, a pleiotropic signalling molecule from Curcuma longa in the treatment of glioblastoma. Eur J Med Chem. 2016;109:23–35.CrossRefGoogle Scholar
  23. 23.
    Purkayastha S, Berliner A, Fernando SS, Ranasinghe B, Ray I, Tariq H, et al. Curcumin blocks brain tumor formation. Brain Res. 2009;1266:130–8.CrossRefGoogle Scholar
  24. 24.
    Priyadarsini KI. The chemistry of curcumin: from extraction to therapeutic agent. Molecules. 2014;19(12):20091–112.CrossRefGoogle Scholar
  25. 25.
    Chiu SS, Lui E, Majeed M, Vishwanatha JK, Ranjan AP, Maitra A, et al. Differential distribution of intravenous curcumin formulations in the rat brain. Anticancer Res. 2011;31(3):907–11.PubMedPubMedCentralGoogle Scholar
  26. 26.
    Wu H, Jiang H, Lu D, Xiong Y, Qu C, Zhao D, et al. Effect of simvastatin on glioma cell proliferation, migration and apoptosis. Neurosurgery. 2009;65(6):1087–97.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Grieb P, Ryba MS, Jagielski J, Gackowski W, Paczkowski P, Chrapusta SJ. Serum cholesterol in cerebral malignancies. J Neuro-Oncol. 1999;41(2):175–80.CrossRefGoogle Scholar
  28. 28.
    Tiwari R, Pathak K. Nanostructured lipid carrier versus solid lipid nanoparticles of simvastatin: comparative analysis of characteristics, pharmacokinetics and tissue uptake. Int J Pharm. 2011;415(1):232–43.CrossRefGoogle Scholar
  29. 29.
    Sengupta R, Sun T, Warrington NM, Rubin JB. Treating brain tumors with PDE4 inhibitors. Trends Pharmacol Sci. 2011;32(6):337–44.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Aguilar MV, Otero C. Frontiers in bioactive compounds, Vol 2. Sharjah: Bentham Science Publishers; 2017.Google Scholar
  31. 31.
    Hansel TT, Tennant RC, Tan AJ, Higgins LA, Neighbour H, Erin EM, et al. Theophylline: mechanism of action and use in asthma and chronic obstructive pulmonary disease. Drugs Today. 2004;40(1):55–69.CrossRefGoogle Scholar
  32. 32.
    PubChem [12-27-2017]. Available from:
  33. 33.
    Pais AACC, Sousa JJS, Vitorino C. Simvastatin delivery: challenges and opportunities. UK: Nova Science Pub Inc; 2015.Google Scholar
  34. 34.
    Hathout RM. Using principal component analysis in studying the transdermal delivery of a lipophilic drug from soft nano-colloidal carriers to develop a quantitative composition effect permeability relationship. Pharm Dev Technol. 2014;19(5):598–604.CrossRefGoogle Scholar
  35. 35.
    Pistone S, Qoragllu D, Smistad G, Hiorth M. Multivariate analysis for the optimization of polysaccharide-based nanoparticles prepared by self-assembly. Colloids Surf B: Biointerfaces. 2016;146:136–43.CrossRefGoogle Scholar
  36. 36.
    Martins S, Tho I, Souto E, Ferreira D, Brandl M. Multivariate design for the evaluation of lipid and surfactant composition effect for optimisation of lipid nanoparticles. Eur J Pharm Sci. 2012;45(5):613–23.CrossRefGoogle Scholar
  37. 37.
    Silva SG, Alves C, Cardoso A, Jurado A, Vale M, Marques EF. Synthesis of gemini surfactants and evaluation of their interfacial and cytotoxic properties: exploring the multifunctionality of serine as headgroup. Eur J Org Chem. 2013;2013:1758–69.CrossRefGoogle Scholar
  38. 38.
    Mendes M, Miranda A, Cova T, Gonçalves L, Almeida AJ, Sousa JJ, et al. Modeling of ultra-small lipid nanoparticle surface charge for targeting glioblastoma. Eur J Pharm Biopharm. 2018;117:255–69.Google Scholar
  39. 39.
    Narang AS, Boddu SH. Excipient applications in formulation design and drug delivery. Cham: Springer International Publishing; 2015.Google Scholar
  40. 40.
    Nanjwade BK, Patel DJ, Udhani RA, Manvi FV. Functions of lipids for enhancement of oral bioavailability of poorly water-soluble drugs. Sci Pharm. 2011;79(4):705–27.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Rowe RC, Sheskey PJ, Quinn ME. Handbook of pharmaceutical excipients. 6th ed. London, Chicago: Pharmaceutical Press, American Pharmacists Association; 2009.Google Scholar
  42. 42.
    Sarkar B, Hardenia S. Microemulsion drug delivery system: for oral bioavailability enhancement of glipizide. J Adv Pharm Educ Res. 2011;1(4):195–200.Google Scholar
  43. 43.
    Macedo JPF, Fernandes LL, Formiga FR, Reis MF, Nagashima Júnior T, Soares LAL, et al. Micro-emultocrit technique: a valuable tool for determination of critical HLB value of emulsions. AAPS PharmSciTech. 2006;7(1):E146–E52.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Gattefossé [12-27-2017]. Available from:
  45. 45.
    IOI Oleo GmbH [12-27-2017]. Available from:
  46. 46.
    ChemSpider [12-27-2017]. Available from:
  47. 47.
    Matsaridou I, Barmpalexis P, Salis A, Nikolakakis I. The influence of surfactant HLB and oil/surfactant ratio on the formation and properties of self-emulsifying pellets and microemulsion reconstitution. AAPS PharmSciTech. 2012;13(4):1319–30.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Sigma-Aldrich [12-27-2017]. Available from:
  49. 49.
    Lipoid GmbH [27-12-2017]. Available from:
  50. 50.
    Bravo González RC, Boess F, Durr E, Schaub N, Bittner B. In vitro investigation on the impact of Solutol HS 15 on the uptake of colchicine into rat hepatocytes. Int J Pharm. 2004;279(1):27–31.CrossRefGoogle Scholar
  51. 51.
    Varmuza K, Filzmoser P. Introduction to multivariate statistical analysis in chemometrics. Boca Raton: CRC Press/Taylor & Francis; 2009.Google Scholar
  52. 52.
    Cova TFGG, Pereira JLGFSC, Pais AACC. Is standard multivariate analysis sufficient in clinical and epidemiological studies? J Biomed Inform. 2013;46(1):75–86.CrossRefGoogle Scholar
  53. 53.
    Jolliffe IT. Principal component analysis. 2nd ed. New York, Berlin: Springer Series in Statistics; 2002.Google Scholar
  54. 54.
    Abdi H, Williams LJ. Principal component analysis. Wiley Interdiscip Rev Comput Stat. 2010;2(4):433–59.CrossRefGoogle Scholar
  55. 55.
    Mutihac L, Mutihac R. Mining in chemometrics. Anal Chim Acta. 2008;612(1):1–18.CrossRefGoogle Scholar
  56. 56.
    Abdi H. Partial least squares regression and projection on latent structure regression (PLS regression). Wiley Interdiscip Rev Comput Stat. 2010;2(1):97–106.CrossRefGoogle Scholar
  57. 57.
    Singh R, Lillard JW. Nanoparticle-based targeted drug delivery. Exp Mol Pathol. 2009;86(3):215–23.CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Mehnert W, Mäder K. Solid lipid nanoparticles: production, characterization and applications. Adv Drug Deliv Rev. 2001;47(2):165–96.CrossRefGoogle Scholar
  59. 59.
    Martins S, Tho I, Ferreira D, Souto E, Brandl M. Physicochemical properties of lipid nanoparticles: effect of lipid and surfactant composition. Drug Dev Ind Pharm. 2011;37(7):815–24.CrossRefGoogle Scholar
  60. 60.
    Gaumet M, Vargas A, Gurny R, Delie F. Nanoparticles for drug delivery: the need for precision in reporting particle size parameters. Eur J Pharm Biopharm. 2008;69(1):1–9.CrossRefGoogle Scholar
  61. 61.
    Müller RH, Jacobs C, Kayser O. Nanosuspensions as particulate drug formulations in therapy: rationale for development and what we can expect for the future. Adv Drug Deliv Rev. 2001;47(1):3–19.CrossRefGoogle Scholar
  62. 62.
    Martins S, Tho I, Reimold I, Fricker G, Souto E, Ferreira D, et al. Brain delivery of camptothecin by means of solid lipid nanoparticles: formulation design, in vitro and in vivo studies. Int J Pharm. 2012;439(1):49–62.CrossRefGoogle Scholar
  63. 63.
    Pathak K, Keshri L, Shah M. Lipid nanocarriers: influence of lipids on product development and pharmacokinetics. Crit Rev Ther Drug Carrier Syst. 2011;28(4):357–93.CrossRefGoogle Scholar
  64. 64.
    Müller RH, Radtke M, Wissing SA. Solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) in cosmetic and dermatological preparations. Adv Drug Deliv Rev. 2002;54:S131–S55.CrossRefGoogle Scholar
  65. 65.
    Abdelbary G, Fahmy RH. Diazepam-loaded solid lipid nanoparticles: design and characterization. AAPS PharmSciTech. 2009;10(1):211–9.CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Hou D, Xie C, Huang K, Zhu C. The production and characteristics of solid lipid nanoparticles (SLNs). Biomaterials. 2003;24(10):1781–5.CrossRefGoogle Scholar
  67. 67.
    Safwat S, Ishak RA, Hathout RM, Mortada ND. Nanostructured lipid carriers loaded with simvastatin: effect of PEG/glycerides on characterization, stability, cellular uptake efficiency and in vitro cytotoxicity. Drug Dev Ind Pharm. 2017;43(7):1112–25.CrossRefGoogle Scholar
  68. 68.
    Miranda A, Blanco-Prieto M, Sousa J, Pais A, Vitorino C. Breaching barriers in glioblastoma. Part I: molecular pathways and novel treatment approaches. Int J Pharm. 2017;531(1):372–88.CrossRefGoogle Scholar
  69. 69.
    Huang W, Tsui GC, Tang C, Yang M. Optimization strategy for encapsulation efficiency and size of drug loaded silica xerogel/polymer core-shell composite nanoparticles prepared by gelation-emulsion method. Polym Eng Sci. 2017;58(5):742–51.Google Scholar
  70. 70.
    Vitorino C, Almeida J, Gonçalves L, Almeida A, Sousa J, Pais A. Co-encapsulating nanostructured lipid carriers for transdermal application: from experimental design to the molecular detail. J Control Release. 2013;167(3):301–14.CrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Jessica Silva
    • 1
    • 2
  • Maria Mendes
    • 1
    • 2
  • Tânia Cova
    • 3
  • João Sousa
    • 1
    • 4
  • Alberto Pais
    • 3
  • Carla Vitorino
    • 1
    • 2
    • 4
    Email author
  1. 1.Faculty of PharmacyUniversity of CoimbraCoimbraPortugal
  2. 2.Centre for Neurosciences and Cell Biology (CNC), Faculty of MedicineUniversity of CoimbraCoimbraPortugal
  3. 3.Coimbra Chemistry Center, Department of ChemistryUniversity of CoimbraCoimbraPortugal
  4. 4.LAQV. REQUIMTE, Group of Pharmaceutical TechnologyPortoPortugal

Personalised recommendations