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Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs

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 quantif...

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Autores principales: Perteghella, Sara, Sottani, Cristina, Coccè, Valentina, Negri, Sara, Cavicchini, Loredana, Alessandri, Giulio, Cottica, Danilo, Torre, Maria Luisa, Grignani, Elena, Pessina, Augusto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631745/
https://www.ncbi.nlm.nih.gov/pubmed/31213025
http://dx.doi.org/10.3390/pharmaceutics11060285
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author Perteghella, Sara
Sottani, Cristina
Coccè, Valentina
Negri, Sara
Cavicchini, Loredana
Alessandri, Giulio
Cottica, Danilo
Torre, Maria Luisa
Grignani, Elena
Pessina, Augusto
author_facet Perteghella, Sara
Sottani, Cristina
Coccè, Valentina
Negri, Sara
Cavicchini, Loredana
Alessandri, Giulio
Cottica, Danilo
Torre, Maria Luisa
Grignani, Elena
Pessina, Augusto
author_sort Perteghella, Sara
collection PubMed
description 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 efficiency (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.
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spelling pubmed-66317452019-08-19 Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs Perteghella, Sara Sottani, Cristina Coccè, Valentina Negri, Sara Cavicchini, Loredana Alessandri, Giulio Cottica, Danilo Torre, Maria Luisa Grignani, Elena Pessina, Augusto Pharmaceutics Article 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 efficiency (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. MDPI 2019-06-17 /pmc/articles/PMC6631745/ /pubmed/31213025 http://dx.doi.org/10.3390/pharmaceutics11060285 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Perteghella, Sara
Sottani, Cristina
Coccè, Valentina
Negri, Sara
Cavicchini, Loredana
Alessandri, Giulio
Cottica, Danilo
Torre, Maria Luisa
Grignani, Elena
Pessina, Augusto
Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
title Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
title_full Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
title_fullStr Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
title_full_unstemmed Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
title_short Paclitaxel-Loaded Silk Fibroin Nanoparticles: Method Validation by UHPLC-MS/MS to Assess an Exogenous Approach to Load Cytotoxic Drugs
title_sort paclitaxel-loaded silk fibroin nanoparticles: method validation by uhplc-ms/ms to assess an exogenous approach to load cytotoxic drugs
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631745/
https://www.ncbi.nlm.nih.gov/pubmed/31213025
http://dx.doi.org/10.3390/pharmaceutics11060285
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