Label-Free Quantification of Nanoencapsulated Piperonyl Esters in Cosmetic Hydrogels Using Raman Spectroscopy

Raman spectroscopy is a well-established technique for the molecular characterisation of samples and does not require extensive pre-analytical processing for complex cosmetic products. As an illustration of its potential, this study investigates the quantitative performance of Raman spectroscopy coupled with partial least squares regression (PLSR) for the analysis of Alginate nanoencapsulated Piperonyl Esters (ANC-PE) incorporated into a hydrogel. A total of 96 ANC-PE samples covering a 0.4% w/w–8.3% w/w PE concentration range have been prepared and analysed. Despite the complex formulation of the sample, the spectral features of the PE can be detected and used to quantify the concentrations. Using a leave-K-out cross-validation approach, samples were divided into a training set (n = 64) and a test set, samples that were previously unknown to the PLSR model (n = 32). The root mean square error of cross-validation (RMSECV) and prediction (RMSEP) was evaluated to be 0.142% (w/w PE) and 0.148% (w/w PE), respectively. The accuracy of the prediction model was further evaluated by the percent relative error calculated from the predicted concentration compared to the true value, yielding values of 3.58% for the training set and 3.67% for the test set. The outcome of the analysis demonstrated the analytical power of Raman to obtain label-free, non-destructive quantification of the active cosmetic ingredient, presently PE, in complex formulations, holding promise for future analytical quality control (AQC) applications in the cosmetics industry with rapid and consumable-free analysis

Estimating the Analytical Performance of Raman Spectroscopy for Quantification of Active Ingredients in Human Stratum Corneum

Confocal Raman microscopy (CRM) has become a versatile technique that can be applied routinely to monitor skin penetration of active molecules. In the present study, CRM coupled to multivariate analysis (namely PLSR—partial least squares regression) is used for the quantitative measurement of an active ingredient (AI) applied to isolated (ex vivo) human stratum corneum (SC), using systematically varied doses of resorcinol, as model compound, and the performance is quantified according to key figures of merit defined by regulatory bodies (ICH, FDA, and EMA). A methodology is thus demonstrated to establish the limit of detection (LOD), precision, accuracy, sensitivity (SEN), and selectivity (SEL) of the technique, and the performance according to these key figures of merit is compared to that of similar established methodologies, based on studies available in literature. First, principal components analysis (PCA) was used to examine the variability within the spectral data set collected. Second, ratios calculated from the area under the curve (AUC) of characteristic resorcinol and proteins/lipids bands (1400–1500 cm−1) were used to perform linear regression analysis of the Raman spectra. Third, cross-validated PLSR analysis was applied to perform quantitative analysis in the fingerprint region. The AUC results show clearly that the intensities of Raman features in the spectra collected are linearly correlated to resorcinol concentrations in the SC (R2 = 0.999) despite a heterogeneity in the distribution of the active molecule in the samples. The Root Mean Square Error of Cross-Validation (RMSECV) (0.017 mg resorcinol/mg SC), The Root Mean Square of Prediction (RMSEP) (0.015 mg resorcinol/mg SC), and R2 (0.971) demonstrate the reliability of the linear regression constructed, enabling accurate quantification of resorcinol. Furthermore, the results have enabled the determination, for the first time, of numerical criteria to estimate analytical performances of CRM, including LOD, precision using bias corrected mean square error prediction (BCMSEP), sensitivity, and selectivity, for quantification of the performance of the analytical technique. This is one step further towards demonstrating that Raman spectroscopy complies with international guidelines and to establishing the technique as a reference and approved tool for permeation studies

Monitoring Dermal Penetration and Permeation Kinetics of Topical Products; the Role of Raman Microspectroscopy

The study of human skin represents an important area of research and development in dermatology, toxicology, pharmacology and cosmetology, in order to assess the effects of exogenous agents, their interaction, their absorption mechanism, and/or their toxicity towards the different cutaneous structures. The processes can be parameterised by mathematical models of diffusion, of varying degrees of complexity, and are commonly measured by Franz cell diffusion, in vitro, and tape stripping, in vitro or in vivo, techniques which are recognised by regulatory bodies for commercialisation of dermally applied products. These techniques do not directly provide chemically specific measurement of the penetration and/or permeation of formulations in situ, however. Raman microspectroscopy provides a non-destructive, non-invasive and chemically specific methodology for in vitro, and in vivo investigations, in-situ, and can provide a powerful alternative to the current gold standard methods approved by regulatory bodies. This review provides an analysis of the current state of art of the field of monitoring dermal penetration and permeation kinetics of topical products, in vitro and in vivo, as well as the regulatory requirements of international guidelines governing them. It furthermore outlines developments in the analysis of skin using Raman microspectroscopy, towards the most recent demonstrations of quantitative monitoring of the penetration and permeation kinetics of topical products in situ, for in vitro and in vivo applications, before discussing the challenges and future perspectives of the field

Développement de nanovecteurs magnétiques de doxorubicine

Our work deals with the conception and the study of magnetic nanoformulations of doxorubicine (DOX). These are aimed to be part of a biocompatible nanovector administered intravenously. Here is presented the functionalization of superparamagnetic iron oxide nanoparticles (SPION) through the adsorption of a DOX-iron(II) complex. This complex and the various stages of functionalization were studied by spectroscopic techniques (UV-visible, fluorescence, surface enhanced Raman scattering spectroscopies). This way of functionalization leads to interesting DOX loadings. We demonstrate the in vitro reversibility of the association between DOX and SPION, and we prove with in vitro cytotoxicity measurements that DOX anticancer activity is preserved. The cell/particles interaction is explored by optical microscopy and microspectrofluorimetry techniques. The experimental results are promising and the DOX-SPION will by used as the active core of therapeutic nanovectors.Ce travail est consacré à la conception et à l étude de nanoformulations magnétiques de doxorubicine (DOX). Celles-ci feront partie d'un nanovecteur compatible avec une administration intraveineuse. Nous présentons une technique de fonctionnalisation de nanoparticules d oxydes de fer superparamagnétiques (SPION) par adsorption d'un complexe DOX-fer(II). Une étude spectroscopique de ce complexe et des étapes de fonctionnalisation a été réalisée (spectroscopies UV-visible, de fluorescence, de diffusion Raman exaltée de surface). Cette voie de fonctionnalisation conduit à des taux de charge intéressants en DOX. Nous démontrons la réversibilité de la liaison SPION/DOX et nous prouvons par des essais de cytotoxicité in vitro que l'association SPION-DOX conserve une activité antitumorale. L'interaction cellule/SPION-DOX est étudiée (microscopie optique et microspectrofluorimétrie confocale). Les résultats expérimentaux sont prometteurs quant à l'utilisation des SPION-DOX comme noyau actif de nanovecteurs thérapeutiques.TOURS-BU Sciences Pharmacie (372612104) / SudocSudocFranceF

La doxorubicine, 40 ans après

TOURS-BU Sciences Pharmacie (372612104) / SudocSudocFranceF

Etude de la stabilité des produits semi-finis et vrac et établissement d'une stratégie de validation des holding time sur un site de production pharmaceutique

Depuis longtemps, il est obligatoire d étudier la stabilité des produits finis et des principes actifs au cours du développement du médicament mais également tout au long de sa durée de vie. Des directives émanant de plusieurs organismes (ICH, OMS, UE ) donnent les exigences à suivre pour la réalisation de ces études. De plus en plus, les textes recommandent également d inclure la stabilité des produits semi-fini et vrac dans les programmes d études de stabilité. Cette démarche peut s inscrire dans le cadre de l implémentation des approches science-based et de la gestion des risques qualité qui sont décrites dans les ICH Q8 et Q9 transposées depuis 2011 dans les BPF. C est cette stratégie qui a été choisie par une entreprise de sous-traitance pharmaceutique pour l étude de la stabilité de ses produits semi-fini et vrac au cours du stockage et la validation de holding time. Trois exemples de validation de holding time seront ainsi présentés dans cette thèse sur trois formes pharmaceutiques différentes : une poudre, un comprimé nu, et une solution de granulation.TOURS-BU Sciences Pharmacie (372612104) / SudocSudocFranceF

Pollution des différentes ressources en eau par les médicaments (risques pour l'environnement et la santé humaine et rôle du pharmacien d'officine dans la collecte des médicaments non utilisés)

Des médicaments chimiquement très divers sont largement consommés à travers le monde à la fois en médecine humaine et vétérinaire. Une fois métabolisés ces médicaments sont éliminés par voie fécale ou urinaire, sous forme de métabolites voire sous forme inchangée. Ainsi les effluents agricoles et piscicoles, les effluents hospitaliers, les effluents domestiques en contiennent. Par ailleurs les médicaments non utilisés sont souvent rejetés avec les ordures ménagères, ou dans les toilettes et les éviers. Cet état de fait est responsable de la présence de résidus médicamenteux dans l environnement et plus particulièrement dans les eaux. Or, les procédés de traitement des effluents en stations d épuration s avèrent insuffisants pour les éliminer, de plus les effluents d élevage ne subissent pas de traitement. On s interroge donc sur leur comportement dans les différentes ressources en eaux et sur les risques potentiels pour la santé humaine ainsi que pour la faune et la flore. Des programmes de recherche et des campagnes de mesures sont menés afin d identifier et d analyser quantitativement et qualitativement les résidus médicamenteux détectés. Le pharmacien d officine a un rôle non négligeable à jouer car il est à même de collecter les médicaments périmés ou non utilisés via le programme CYCLAMED. Ces médicaments sont alors détruits. Il est d une importance capitale de sensibiliser le public au bien-être de la planète et déjà les inciter à ramener leurs médicaments non utilisés ou périmés chez leur pharmacien est un pas vers une moindre contamination de l environnementTOURS-BU Sciences Pharmacie (372612104) / SudocSudocFranceF

Recent advances in theranostic nanocarriers of doxorubicin based on iron oxide and gold nanoparticles.

International audienceHybrid (organic/inorganic) nanoparticles emerged as a simple solution to build "theranostic" systems. Due to their physical properties, superparamagnetic iron oxide nanoparticles (SPIONs) and plasmonic gold nanoparticles (Au-NPs) are extensively studied as a part of diagnostic and therapeutic strategies in cancer treatments. They can be used as agents for in vitro or in vivo imaging, for magnetic drug targeting and/or thermal therapy. Their functionalization with organic shells enhances their potential performance in tumor targeting and drug delivery. The advances in such hybrid nanocarriers are well illustrated with the example of the anticancer drug doxorubicin (DOX). The aim of this review is to give a multidisciplinary overview of such smart nanosystems loaded with DOX, based on examples taken from recent publications. From a physico-chemical point of view, we discuss the choices for the strategies for loading DOX and the consequences on drug release. From a biological point of view, we analyze the in vitro and in vivo assays concerning tumor imaging, targeted drug delivery and anticancer efficiency. Future opportunities and challenges are also addressed

Versatile electrostatically assembled polymeric siRNA nanovectors: Can they overcome the limits of siRNA tumor delivery?

International audienceThe application of small interfering RNA (siRNA) cancer therapeutics is limited by several extra- and intracellular barriers including the presence of ribonucleases that degrade siRNA, the premature clearance, the impermeability of the cell membrane, or the difficulty to escape endo-lysosomal degradation. Therefore, several delivery systems have emerged to overcome these limitations and to successfully deliver siRNA to the tumor site. This review is focused on polymer-based siRNA nanovectors which exploit the negative charge of siRNA, representing a major challenge for siRNA delivery, to their advantage by loading siRNA via electrostatic assembly. These nanovectors are easy to prepare and to adapt for an optimal gene silencing efficiency. The ability of electrostatically assembled polymeric siRNA nanovectors (EPSN) to improve the half-life of siRNA, to favor the specificity of the delivery and the accumulation in tumor and to enhance the cellular uptake and endosomal escape for an efficient siRNA delivery will be discussed. Finally, the influence of the versatility of the structure of these nanovectors on the protein down-regulation will be evaluated