The aim of this work was to load an anticancer drug, paclitaxel (PTX), on Silk Fibroin Nanoparticles (SFNs) by using an exogenous approach. SFNs were produced, freeze-dried and then loaded with PTX. An exogenous method allowed us to reduce both drug loss and environmental impact. In order to quantify PTX loaded in SFNs, a simple and reliable method using reversed phase liquid chromatography coupled to tandem mass spectrometry (rp-UHPLC-MS/MS) was developed. This methodology was validated by the determination of spiked QC samples in three consecutive days. Good accuracy and precision of the method were obtained, while the intra-day and inter-day precisions were less than 10.3%. For PTX, the limit of quantitation (LOQ) was 5.0 ng/mL. Recovery from the matrix (SFNs-PTX pellets) was calculated (81.2% at LOQ value) as PTX was entrapped in a new matrix like the polymer silk fibroin-based. This method was successfully applied to determine the encapsulation effciency (1.00 ± 0.19%) and the nanoparticle loading (0.12 ± 0.02% w/w). The in vitro anticancer activity of SFNs-PTX was tested against CFPAC-1 cancer cells; results demonstrated a very high cytotoxic activity of SFNs-PTX, with a dose dependent inhibition of CFPAC-1 proliferation, confirmed by the IC50 value of 3450 ± 750 ng/mL.
Paclitaxel-loaded silk fibroin nanoparticles: Method validation by UHPLC-MS/MS to assess an exogenous approach to load cytotoxic drugs / S. Perteghella, C. Sottani, V. Cocce', S. Negri, L. Cavicchini, G. Alessandri, D. Cottica, M.L. Torre, E. Grignani, A. Pessina. - In: PHARMACEUTICS. - ISSN 1999-4923. - 11:6(2019 Jun 17), pp. 285.1-285.15.
|Titolo:||Paclitaxel-loaded silk fibroin nanoparticles : Method validation by UHPLC-MS/MS to assess an exogenous approach to load cytotoxic drugs|
|Parole Chiave:||Method validation; Nanoparticles; Paclitaxel; Silk fibroin; UHPLC-MS/MS|
|Settore Scientifico Disciplinare:||Settore BIO/13 - Biologia Applicata|
|Data di pubblicazione:||17-giu-2019|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.3390/pharmaceutics11060285|
|Appare nelle tipologie:||01 - Articolo su periodico|