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Non-viral delivery using cationic solid lipid nanoparticles (SLNs) represents a useful strategy to introduce large DNA and RNA molecules to target cells. A careful selection of components and their amounts is critical to improve transfection efficiency. In this work, a selected and optimized formulation of SLNs was used to efficiently transfect circular DNA and linear RNA molecules into cells. We characterized the main physicochemical characteristics and binding capabilities of these SLNs and show that they deliver DNA and RNA molecules into cells where they display full bioactivity at nontoxic concentrations using fluorescence-and luminescence-based methodologies. Hence, we established a novel and simple SLN formulation as a powerful tool for future therapeutic use. (C) 2016 Elsevier B.V. All rights reserved., This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (grant number BFU2014-54660-R) and the Andalusian Government (Excellence Project BIO-2515/2012) to C.S. and from the Spanish Ministry of Economy and Competitiveness (grant number BFU2013-44660-R) and the Andalusian Government (Excellence Project CTS-6587) to C.H.M. Support from the European Region Development Fund (ERDF [FEDER]), is also acknowledged.

A new and simple reverse phase high performance liquid chromatography method was developed for the simultaneous determination of the major compounds of skin-whitening pharmaceutical cream, i.e., two active pharmaceutical ingredients: hydroquinone and kojic acid, two sunscreens: octinoxate and avobenzone and two antioxidants: butylated hydroxyanisole and butylated hydroxytoluene. Separation was achieved on a Zorbax SB-Phenyl (250 × 4.6 mm; 5 μm) column, using a mobile phase consisting of water with 0.1% acetic acid and acetonitrile, in a dual-mode gradient for 14 minutes. The column was maintained at 40 °C and detection was carried out at 230 nm by using a diode-array detector. The extraction procedure was studied by a design of experiments demonstrating its suitability. The structure of an additional unknown peak eluting in all chromatograms in the same proportion was identified by HPLC/MS as the keto tautomer of avobenzone. The method developed was successfully validated in accordance with ICH guidelines with regard to selectivity, linearity, accuracy, precision and robustness. Selectivity was found to be satisfactory. The linear regression analysis data for all compounds showed a good linear relationship with r2 > 0.999 in the concentration range from 50% to 120% of the label claimed for each compound. The percentage RSD for precision and accuracy of the method was found to be less than 2% for all compounds. The proposed method was applied for the determination of six major compounds in skin-whitening cream prototypes developed during the formulation development stage.

Background: Cationic solid lipid nanoparticles (SLNs) have been given considerable attention for therapeutic nucleic acid delivery owing to their advantages over viral and other nanoparticle delivery systems. However, poor delivery efficiency and complex formulations hinder the clinical translation of SLNs. Aim: The aim of this study was to formulate and characterize SLNs incorporating the cholesterol derivative cholesteryl oleate to produce SLN-nucleic acid complexes with reduced cytotoxicity and more efficient cellular uptake. Methods: Five cholesteryl oleate-containing formulations were prepared. Laser diffraction and laser Doppler microelectrophoresis were used to evaluate particle size and zeta potential, respectively. Nanoparticle morphology was analyzed using electron microscopy. Cytotoxicity and cellular uptake of lipoplexes were evaluated using flow cytometry and fluorescence microscopy. The gene inhibition capacity of the lipoplexes was assessed using siRNAs to block constitutive luciferase expression. Results: We obtained nanoparticles with a mean diameter of approximately 150-200 nm in size and zeta potential values of 25-40 mV. SLN formulations with intermediate concentrations of cholesteryl oleate exhibited good stability and spherical structures with no aggregation. No cell toxicity of any reference SLN was observed. Finally, cellular uptake experiments with DNA- and RNA-SLNs were performed to select one reference with superior transient transfection efficiency that significantly decreased gene activity upon siRNA complexation. Conclusion: The results indicate that cholesteryl oleate-loaded SLNs are a safe and effective platform for nonviral nucleic acid delivery., We are grateful to many colleagues for their helpful suggestions, critical discussions, and comments. We thank Jose Alcami and Mayte Coiras (National Microbiology Center, Majadahonda, Madrid, Spain) for the TZM-bl cells. This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (grant number BFU2014-54660-R) and the Andalusian Government (Excellence Project BIO-2515/2012) to CS and from the Spanish Ministry of Economy and Competitiveness (grant number BFU2016-79699-P) to CHM. Support from the European Region Development Fund (ERDF [FEDER]) is also acknowledged. MSP was supported by a fellowship from the Spanish Ministry of Education (FPU Program [Training Program for Academic Staff]).

Risk analysis tools can be applied in the early stages of the research, development, and innovation of pharmaceutical drugs. We used a risk ranking and filtering method to optimize time resources in internal audits of project development documents in order to ensure traceability in a university pilot plant. Data gathered during audits undertaken over a 14 month period were classified according to risk factors at several levels. Consequently, time resources for this type of internal audit can be optimised by focusing on aspects that are objectively determined to be critical to traceability. In addition, quality can be ensured by paying more attention to the most critical aspects, rather than the most commonly observed findings in historical data. LAY ABSTRACT: One of the most important issue for a research, development, and innovation centre is the traceability of both experimental tasks and their documentation. This study describes an analysis of the findings revealed during internal audits carried out over a 14 month period, in order to establish its criticality and to highlight those important aspects to consider in future audits, while saving both personnel and time resources to carry out internal audits on development of projects.