Influence of lipid concentration on encapsulation and particle size in the development of liposomal irinotecan


  • A. V. Stadnichenko National University of Pharmacy,
  • Y. M. Krasnopolsky National Technical Univercity «Kharkiv Politechnical Institute»,
  • T. G. Yarnykh National University of Pharmacy,



liposomes, irinotecan, lipid bilayers, phosphatidylcholine, cholesterol, high pressure homogenization


Purpose. For creation of liposomal irinotecan form, the influence of lipid concentration on the encapsulation of the active substance and nanoparticle size  was necessary to investigate.

Materials and methods. We used «chemical gradient" method for liposomes formulation, in the variety of “pH gradient”. Ammonium citrate at pH 2.5 was used as internal buffer. Lipid film was obtained, by evaporation technics with further high pressure homogenization with Microfluidics Microfluidizer M-110P apparatus. “Chemical gradient” was created by ultrafiltration, with “Minim 2” apparatus. Ultrafiltration cartridge with an upper cut-off 30 kDa was used. Encapsulation was measured using HPLC methods developed in variant of gel chromatography, with Shimadzu LC-20 instrument. The particle size was measured by laser diffraction method withe “Zetasizer Nano ZS” instrument.

Results. For the preparation of liposomes was applied a constant lipid ratio with varying of total lipid concentration. The ratio of lipids in the experiment was a phosphatidylcholine / cholesterol 80/20 % by weight. A total concentration was investigated in range from 10 mg/ml to 30 mg/ml. The number of extrusion cycles consisted from 3 cycles at 1500 bar, in case of 10 mg/ml concentration, to 17 cycles at 1500 bar, in case of 30 mg/ml concentration.

Conclusions. It was shown that the lipid concentration from 25 mg/ml led to particles formation with size more than 5000 nm, and it was not possible to reduce them by high pressure homogenization method. It was proven that the most optimal, in terms of technology and the final characteristics of liposomes, was lipids in concentration 20 mg/ml. The degree of encapsulation in this case was 82 ± 0.98 %. The size of the liposomes was 106 nm, 5000 nm particles were absent.


Frenzel, M., & Steffen-Heins, A. (2015) Impact of quercetin and fish oil encapsulation on bilayer membrane and oxidation stability of liposomes. Food Chemistry, 185, 48–57. doi: 10.1016/j.foodchem.2015.03.121.

Liau, J. J., Hook, S., Prestidge, C. A., & Barnes, T. J. (2015) A lipid based multi-compartmental system: Liposomes-in-double emulsion for oral vaccine delivery. European Journal of Pharmaceutics and Biopharmaceutics, 97, 15–21. doi: 10.1016/j.ejpb.2015.09.018.

Rizzitelli, S., Giustetto, P., Faletto, D., Delli Castelli, D., Aime, S., & Terreno, E. (2016) The release of Doxorubicin from liposomes monitored by MRI and triggered by a combination of US stimuli led to a complete tumor regression in a breast cancer mouse model. Journal of Controlled Release, 230, 57–63. doi: 10.1016/j.jconrel.2016.03.040.

Ron-Doitch, S., Sawodny, B., Kühbacher, A., David, M. M., Samanta, A., Phopase, J., et al. (2016) Reduced cytotoxicity and enhanced bioactivity of cationic antimicrobial peptides liposomes in cell cultures and 3D epidermis model against HSV. Journal of Controlled Release, 10(229), 163–171. doi: 10.1016/j.jconrel.2016.03.025.

Kulkarni, P. R., Yadav, J. D., & Vaida, K. A. (2011) Liposomes: a novel drug delivery system. International Journal of Current Pharmaceutical Research, 3(2), 32–41.

Li, Y., Liu, R., Yang, J., Shi, Y., Ma, G., Zhang, Z., & Zhang, X. (2015) Enhanced retention and anti-tumor efficacy of liposomes by changing their cellular uptake and pharmacokinetics behavior. Biomaterials, 41, 1–14. doi: 10.1016/j.biomaterials.2014.11.010.

Torchilin, V. (2011) Tumor delivery of macromolecular drugs based on the EPR effect. Advanced Drug Delivery Reviews, 63, 131–135. doi: 10.1016/j.addr.2010.03.011.

Stadnichenko, A. V., Krasnopoljskij, Y. M., & Shvets, V. I. (2014) Tekhnologiya polucheniya liposomal'nykh form irinotekana (obzor). [Technology for production of liposomal irinotecan (a review)]. Biofarmacevticheskij zhurnal, 6(6), 3–9. [in Russian].

West-Ward Pharmaceuticals Corp. (2016) Irinotecan prescribing information. Retrieved from: irinotecan.html.

Paulіk, A., Grim, J., & Filip, S. (2012) Predictors of Irinotecan toxicity and efficancy in treatment of metastatic colorectal cancer. Acta Medica, 55(4), 153–159. doi: 10.14712/18059694.2015.39.

How to Cite

Stadnichenko AV, Krasnopolsky YM, Yarnykh TG. Influence of lipid concentration on encapsulation and particle size in the development of liposomal irinotecan. Current issues in pharmacy and medicine: science and practice [Internet]. 2017Feb.20 [cited 2024May24];(1). Available from:



Pharmaceutical manufacturing