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Open AccessArticle

Insights on Ultrafiltration-Based Separation for the Purification and Quantification of Methotrexate in Nanocarriers

1
LAQV, REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira n 228, 4050-313 Porto, Portugal
2
Escola Superior de Saúde, Instituto Politécnico do Porto, 4200-072 Porto, Portugal
3
Centro de Espetrometria de Massa, Departamento de Química & QOPNA, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal
4
Departamento de Química & CESAM & ECOMARE, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal
*
Author to whom correspondence should be addressed.
Molecules 2020, 25(8), 1879; https://doi.org/10.3390/molecules25081879
Received: 15 March 2020 / Revised: 6 April 2020 / Accepted: 14 April 2020 / Published: 18 April 2020
(This article belongs to the Section Analytical Chemistry)
The evaluation of encapsulation efficiency is a regulatory requirement for the characterization of drug delivery systems. However, the difficulties in efficiently separating nanomedicines from the free drug may compromise the achievement of accurate determinations. Herein, ultrafiltration was exploited as a separative strategy towards the evaluation of methotrexate (MTX) encapsulation efficiency in nanostructured lipid carriers and polymeric nanoparticles. The effect of experimental conditions such as pH and the amount of surfactant present in the ultrafiltration media was addressed aiming at the selection of suitable conditions for the effective purification of nanocarriers. MTX-loaded nanoparticles were then submitted to ultrafiltration and the portions remaining in the upper compartment of the filtering device and in the ultrafiltrate were collected and analyzed by HPLC-UV using a reversed-phase (C18) monolithic column. A short centrifugation time (5 min) was suitable for establishing the amount of encapsulated MTX in nanostructured lipid carriers, based on the assumption that the free MTX concentration was the same in the upper compartment and in the ultrafiltrate. The defined conditions allowed the efficient separation of nanocarriers from the free drug, with recoveries of >85% even when nanoparticles were present in cell culture media and in pig skin surrogate from permeation assays. View Full-Text
Keywords: drug delivery; encapsulation efficiency; permeation; separative methods; ultrafiltration drug delivery; encapsulation efficiency; permeation; separative methods; ultrafiltration
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MDPI and ACS Style

Marques, S.S.; Ramos, I.I.; Fernandes, S.R.; Barreiros, L.; Lima, S.A.C.; Reis, S.; Domingues, M.R.M.; Segundo, M.A. Insights on Ultrafiltration-Based Separation for the Purification and Quantification of Methotrexate in Nanocarriers. Molecules 2020, 25, 1879.

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