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  1. Article: Monitoring doxorubicin cellular uptake and trafficking using in vitro Raman microspectroscopy: short and long time exposure effects on lung cancer cell lines

    Farhane, Zeineb / Franck Bonnier / Hugh J. Byrne

    Analytical and bioanalytical chemistry. 2017 Feb., v. 409, no. 5

    2017  

    Abstract: Raman microspectroscopy is a non-invasive, in vitro analytical tool which is being increasingly explored for its potential in clinical applications and monitoring the uptake, mechanism of action and cellular interaction at a molecular level of ... ...

    Abstract Raman microspectroscopy is a non-invasive, in vitro analytical tool which is being increasingly explored for its potential in clinical applications and monitoring the uptake, mechanism of action and cellular interaction at a molecular level of chemotherapeutic drugs, ultimately as a potential label-free preclinical screening and companion diagnostic tool. In this study, doxorubicin (DOX), a “gold standard” chemotherapeutic drug, is employed as a model in the in vitro lung cancer cell line A549 in order to demonstrate the potential of Raman microspectroscopy to screen and identify spectroscopic markers of its trafficking and mechanism of action. Confocal laser scanning microscopy (CLSM) was used in parallel to illustrate the uptake and subcellular localisation, and cytotoxicity assays were employed to establish the toxicity profiles for early and late exposure times of A549 to DOX. Multivariate statistical analysis, consisting of principal components analysis (PCA), partial least squares regression (PLSR) and independent component analysis (ICA), was used to elucidate the spectroscopic signatures associated with DOX uptake and subcellular interaction. Raman spectroscopic profiling illustrates both drug kinetics and its pharmacodynamics in the cell and associated biochemical changes, demonstrating that DOX is mainly localised in the nuclear area, saturating the nucleolus first, within ~6 h of exposure, before the surrounding nuclear areas after ~12 h, and only accumulates in the cytoplasm after 48 h. PLSR over varying time intervals enables identification of DOX–DNA binding at early stages of exposure (0–12 h), while regression over longer time periods (24–72 h) reveals spectroscopic signatures associated with the metabolic cellular response. Graphical Abstract Subcellular uptake of doxorubicin, and changes in biomolecular signatures in the nucleolus, as monitored by Raman spectroscopy
    Keywords Raman spectroscopy ; cell nucleolus ; confocal laser scanning microscopy ; cytoplasm ; cytotoxicity ; diagnostic techniques ; doxorubicin ; drug therapy ; exposure duration ; independent component analysis ; least squares ; lung neoplasms ; mechanism of action ; models ; monitoring ; pharmacodynamics ; pharmacokinetics ; principal component analysis ; screening
    Language English
    Dates of publication 2017-02
    Size p. 1333-1346.
    Publishing place Springer Berlin Heidelberg
    Document type Article
    ISSN 1618-2642
    DOI 10.1007/s00216-016-0065-0
    Database NAL-Catalogue (AGRICOLA)

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  2. Article ; Online: Estimating the Analytical Performance of Raman Spectroscopy for Quantification of Active Ingredients in Human Stratum Corneum

    Hichem Kichou / Emilie Munnier / Yuri Dancik / Kamilia Kemel / Hugh J. Byrne / Ali Tfayli / Dominique Bertrand / Martin Soucé / Igor Chourpa / Franck Bonnier

    Molecules, Vol 27, Iss 2843, p

    2022  Volume 2843

    Abstract: 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 ... ...

    Abstract 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 (R 2 = 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 R 2 (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 ...
    Keywords stratum corneum ; high performance liquid chromatography ; confocal Raman spectroscopy ; resorcinol ; quantification ; Organic chemistry ; QD241-441
    Subject code 290
    Language English
    Publishing date 2022-04-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Comparison of Vibrational Spectroscopic Techniques for Quantification of Water in Natural Deep Eutectic Solvents

    Suha Elderderi / Pierre-Yves Sacré / Laura Wils / Igor Chourpa / Abdalla A. Elbashir / Philippe Hubert / Hugh J. Byrne / Leslie Boudesocque-Delaye / Eric Ziemons / Franck Bonnier

    Molecules, Vol 27, Iss 4819, p

    2022  Volume 4819

    Abstract: Vibrational spectroscopic techniques, i.e., attenuated total reflectance infrared (ATR-IR), near infrared spectroscopy (NIRS) and Raman spectroscopy (RS), coupled with Partial Least Squares Regression (PLSR), were evaluated as cost-effective label-free ... ...

    Abstract Vibrational spectroscopic techniques, i.e., attenuated total reflectance infrared (ATR-IR), near infrared spectroscopy (NIRS) and Raman spectroscopy (RS), coupled with Partial Least Squares Regression (PLSR), were evaluated as cost-effective label-free and reagent-free tools to monitor water content in Levulinic Acid/L-Proline (LALP) (2:1, mol/mol) Natural Deep Eutectic Solvent (NADES). ATR-IR delivered the best outcome of Root Mean Squared Error (RMSE) of Cross-Validation (CV) = 0.27% added water concentration, RMSE of Prediction (P) = 0.27% added water concentration and mean % relative error = 2.59%. Two NIRS instruments (benchtop and handheld) were also compared during the study, respectively yielding RMSECV = 0.35% added water concentration, RMSEP = 0.56% added water concentration and mean % relative error = 5.13% added water concentration, and RMECV = 0.36% added water concentration, RMSEP = 0.68% added water concentration and mean % relative error = 6.23%. RS analysis performed in quartz cuvettes enabled accurate water quantification with RMECV = 0.43% added water concentration, RMSEP = 0.67% added water concentration and mean % relative error = 6.75%. While the vibrational spectroscopic techniques studied have shown high performance in relation to reliable determination of water concentration, their accuracy is most likely related to their sensitivity to detect the LALP compounds in the NADES. For instance, whereas ATR-IR spectra display strong features from water, Levulinic Acid and L-Proline that contribute to the PLSR predictive models constructed, NIRS and RS spectra are respectively dominated by either water or LALP compounds, representing partial molecular information and moderate accuracy compared to ATR-IR. However, while ATR-IR instruments are common in chemistry and physics laboratories, making the technique readily transferable to water quantification in NADES, Raman spectroscopy offers promising potential for future development for in situ, sample withdrawal-free analysis for high throughput ...
    Keywords label-free water quantification ; natural deep eutectic solvent ; partial least squares regression ; attenuated total reflection infrared ; Raman spectroscopy ; near infrared spectroscopy ; Organic chemistry ; QD241-441
    Language English
    Publishing date 2022-07-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: In Situ Water Quantification in Natural Deep Eutectic Solvents Using Portable Raman Spectroscopy

    Suha Elderderi / Laura Wils / Charlotte Leman-Loubière / Hugh J. Byrne / Igor Chourpa / Cécile Enguehard-Gueiffier / Emilie Munnier / Abdalla A. Elbashir / Leslie Boudesocque-Delaye / Franck Bonnier

    Molecules, Vol 26, Iss 5488, p

    2021  Volume 5488

    Abstract: Raman spectroscopy is a label-free, non-destructive, non-invasive analytical tool that provides insight into the molecular composition of samples with minimum or no sample preparation. The increased availability of commercial portable Raman devices ... ...

    Abstract Raman spectroscopy is a label-free, non-destructive, non-invasive analytical tool that provides insight into the molecular composition of samples with minimum or no sample preparation. The increased availability of commercial portable Raman devices presents a potentially easy and convenient analytical solution for day-to-day analysis in laboratories and production lines. However, their performance for highly specific and sensitive analysis applications has not been extensively evaluated. This study performs a direct comparison of such a commercially available, portable Raman system, with a research grade Raman microscope system for the analysis of water content of Natural Deep Eutectic Solvents (NADES). NADES are renewable, biodegradable and easily tunable “green” solvents, outcompeting existing organic solvents for applications in extraction from biomass, biocatalysis, and nanoparticle synthesis. Water content in NADES is, however, a critical parameter, affecting their properties, optimal use and extraction efficiency. In the present study, portable Raman spectroscopy coupled with Partial Least Squares Regression (PLSR) is investigated for rapid determination of water content in NADES samples in situ, i.e., directly in glassware. Three NADES systems, namely Betaine Glycerol (BG), Choline Chloride Glycerol (CCG) and Glucose Glycerol (GG), containing a range of water concentrations between 0% ( w / w ) and 28.5% ( w / w ), were studied. The results are directly compared with previously published studies of the same systems, using a research grade Raman microscope. PLSR results demonstrate the reliability of the analysis, surrendering R 2 values above 0.99. Root Mean Square Errors Prediction (RMSEP) of 0.6805%, 0.9859% and 1.2907% w / w were found for respectively unknown CCG, BG and GG samples using the portable device compared to 0.4715%, 0.3437% and 0.7409% w / w previously obtained by analysis in quartz cuvettes with a Raman confocal microscope. Despite the relatively higher values of RMSEP observed, the comparison of the percentage of relative errors in the predicted concentration highlights that, overall, the portable device delivers accuracy below 5%. Ultimately, it has been demonstrated that portable Raman spectroscopy enables accurate quantification of water in NADES directly through glass vials without the requirement for sample withdrawal. Such compact instruments provide solvent and consumable free analysis for rapid analysis directly in laboratories and for non-expert users. Portable Raman is a promising approach for high throughput monitoring of water content in NADES that can support the development of new analytical protocols in the field of green chemistry in research and development laboratories but also in the industry as a routine quality control tool.
    Keywords deep eutectic solvent ; portable Raman spectroscopy ; label free water quantification ; in situ analysis ; partial least squares regression ; Organic chemistry ; QD241-441
    Subject code 290
    Language English
    Publishing date 2021-09-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Confocal Raman Spectroscopic Imaging for Evaluation of Distribution of Nano-Formulated Hydrophobic Active Cosmetic Ingredients in Hydrophilic Films

    Louise Van Gheluwe / Emilie Munnier / Hichem Kichou / Kamilia Kemel / Frédéric Mahut / Marylène Vayer / Christophe Sinturel / Hugh J. Byrne / Florent Yvergnaux / Igor Chourpa / Franck Bonnier

    Molecules, Vol 26, Iss 7440, p

    2021  Volume 7440

    Abstract: Film-forming systems are highly relevant to the topical administration of active ingredients (AI) to the body. Enhanced contact with the skin can increase the efficacy of delivery and penetration during prolonged exposure. However, after the evaporation ... ...

    Abstract Film-forming systems are highly relevant to the topical administration of active ingredients (AI) to the body. Enhanced contact with the skin can increase the efficacy of delivery and penetration during prolonged exposure. However, after the evaporation of volatile solvents to form a thin film, the distribution of the ingredient should remain homogenous in order to ensure the effectiveness of the formula. This is especially critical for the use of hydrophobic molecules that have poor solubility in hydrophilic films. In order to address this concern, hydroxyphenethyl esters (PHE) of Punica granatum seed oil were prepared as a nanosuspension stabilised by poloxamers (NanoPHE). NanoPHE was then added to a formulation containing polyvinyl alcohol (PVA) as a film forming agent, Glycerol as a plasticiser and an antimicrobial agent, Sepicide TM HB. Despite their reliability, reference methods such as high-performance liquid chromatography are increasingly challenged due to the need for consumables and solvents, which is contrary to current concerns about green industry in the cosmetics field. Moreover, such methods fail to provide spatially resolved chemical information. In order to investigate the distribution of ingredients in the dried film, Confocal Raman imaging (CRI) coupled to Non-negatively Constrained Least Squares (NCLS) analysis was used. The reconstructed heat maps from a range of films containing systematically varying PHE concentrations highlighted the changes in spectral contribution from each of the ingredients. First, using NCLS scores it was demonstrated that the distributions of PVA, Glycerol, Sepicide TM HB and PHE were homogenous, with respective relative standard deviations (RSD) of 3.33%, 2.48%, 2.72% and 6.27%. Second, the respective relationships between ingredient concentrations in the films and their Raman responses, and the spectral abundance were established. Finally, a model for absolute quantification for PHE was be constructed using the percentage of spectral abundance. The prepared % w ...
    Keywords active cosmetic ingredients ; nanosuspension ; polyvinyl alcohol ; film ; confocal Raman imaging ; multivariate analysis ; Organic chemistry ; QD241-441
    Subject code 333
    Language English
    Publishing date 2021-12-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article: Quantitative analysis of curcumin-loaded alginate nanocarriers in hydrogels using Raman and attenuated total reflection infrared spectroscopy

    Miloudi, Lynda / Franck Bonnier / Dominique Bertrand / Hugh J. Byrne / Xavier Perse / Igor Chourpa / Emilie Munnier

    Analytical and bioanalytical chemistry. 2017 July, v. 409, no. 19

    2017  

    Abstract: Core-shell nanocarriers are increasingly being adapted in cosmetic and dermatological fields, aiming to provide an increased penetration of the active pharmaceutical or cosmetic ingredients (API and ACI) through the skin. In the final form, the ... ...

    Abstract Core-shell nanocarriers are increasingly being adapted in cosmetic and dermatological fields, aiming to provide an increased penetration of the active pharmaceutical or cosmetic ingredients (API and ACI) through the skin. In the final form, the nanocarriers (NC) are usually prepared in hydrogels, conferring desired viscous properties for topical application. Combined with the high chemical complexity of the encapsulating system itself, involving numerous ingredients to form a stable core and quantifying the NC and/or the encapsulated active without labor-intensive and destructive methods remains challenging. In this respect, the specific molecular fingerprint obtained from vibrational spectroscopy analysis could unambiguously overcome current obstacles in the development of fast and cost-effective quality control tools for NC-based products. The present study demonstrates the feasibility to deliver accurate quantification of the concentrations of curcumin (ACI)-loaded alginate nanocarriers in hydrogel matrices, coupling partial least square regression (PLSR) to infrared (IR) absorption and Raman spectroscopic analyses. With respective root mean square errors of 0.1469 ± 0.0175% w/w and 0.4462 ± 0.0631% w/w, both approaches offer acceptable precision. Further investigation of the PLSR results allowed to highlight the different selectivity of each approach, indicating only IR analysis delivers direct monitoring of the NC through the quantification of the Labrafac®, the main NC ingredient. Raman analyses are rather dominated by the contribution of the ACI which opens numerous perspectives to quantify the active molecules without interferences from the complex core-shell encapsulating systems thus positioning the technique as a powerful analytical tool for industrial screening of cosmetic and pharmaceutical products. Graphical abstract Quantitative analysis of encapuslated active molecules in hydrogel-based samples by means of infrared and Raman spectroscopy
    Keywords Raman spectroscopy ; absorption ; alginates ; cost effectiveness ; curcumin ; drugs ; encapsulation ; hydrogels ; infrared spectroscopy ; ingredients ; least squares ; monitoring ; nanocarriers ; quality control ; quantitative analysis ; screening ; spectral analysis ; topical application
    Language English
    Dates of publication 2017-07
    Size p. 4593-4605.
    Publishing place Springer Berlin Heidelberg
    Document type Article
    ISSN 1618-2642
    DOI 10.1007/s00216-017-0402-y
    Database NAL-Catalogue (AGRICOLA)

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