Optical microcavities exhibit high resonance quality, so that, they are of key interest for the design of low-threshold lasers or for achieving strong coupling regime. But, such systems support modes whose the volume remain diffraction limited.In this manuscript, we are interested in their plasmonic counterparts because they support confined modes at the sub-wavelength scale. First, we study an in-plane plasmonic cavity which is the transposition of 1D optical cavity to surface wave. We characterize the cavity by measuring the fluorescence lifetime of dye molecules deposited inside.Then, we are interested in 3-dimension mode confinement achieved by spherical metal nanoparticles. We discuss on the definition of the mode volume used in cavity quantum electrodynamic and based on the calculation of energy confinement around the particle. We also simulate the fluorescence enhancement of rare-earth ions embedded inside core-shell plasmonic particles. Finally, we disturb the photodynamic emission of a single-photon source by puttingthe extremity of a plasmonic tip nearby the emitter

Η φύση της οπτικής διέγερσης και ο βαθμός της ενδομοριακής μεταφοράς του φορτίου (Intramolecular Charge Transfer, ICT), καθώς επίσης η δυναμική των διεγερμένων στάθμεων, σε δύο νέες οργανικές δομές μορίων , D-π-A (γραμμικό μόριο) και DΑ3 (δενδριμερές - αστεροειδές μόριο με επίκεντρο τον δότη ηλεκτρονίων) έχουν διερευνηθεί με femtosecond φασματοσκοπία χρονικής ανάλυσης (απόσβεση του φθορισμού τους) σε δύο διαφορετικούς διαλύτες, το τολουόλιο (με ασθενή πολικότητα) και το τετραϋδροφουράνιο (με μέτρια πολικότητα). Τα πειραματικά αποτελέσματα με βάση τη σύγκριση των μορίων της ίδιας δομής (DA ή DA3) με προοδευτική αλλαγή των αποδεκτών ηλεκτρονίων (από τους ασθενέστερους προς τους ισχυρότερους) στον ίδιο διαλύτη έδειξαν ότι οι οργανικές δομές με υποκαταστάτες τους ισχυρότερους αποδέκτες ηλεκτρονίων είχαν γρηγορότερη απόσβεση του φθορισμού τους και συνεπώς ισχυρότερη ενδομοριακή μεταφορά του φορτίου. Επιπλέον, η σύγκριση των δύο δομών έδειξε ότι τα δενδριμερή μόρια γενικά παρουσίαζαν ισχυρότερη ενδομοριακή μεταφορά του φορτίου. Επίσης τα πειραματικά αποτελέσματα έδειξαν ότι κατά τη σύγκριση των δύο διαλυτών οι αλληλεπιδράσεις διπόλου - διπόλου μεταξύ της διαλυμένης ουσίας και του διαλύτη επιδρά στις φασματικές ιδιότητες των μορίων μειώνοντας την ενέργεια της διεγερμένης στάθμης αφού παρατηρήθηκε ερυθρή μετατόπιση Stoke (red shift Stoke) των φασμάτων φθορισμού των μορίων και ακολούθως οι δομές με τους ισχυρότερους αποδέκτες ηλεκτρονίων παρουσίαζαν μεγαλύτερη απόσβεση του φθορισμού. / The optic excitement and the degree of Intramolecular Charge Transfer (ICT), as well as the dynamics of excited states, two new organic structures of molecules, D-pi-A (linear molecule) and DA3 (dendrimers – molecule with center an electron donor) have been investigated with femtosecond time resolved spectroscopy in two different solvents, toluene (with weak polarity) and tetrahydrofuran (moderate polarity). The experimental results by comparing the molecules of the same structure (DA or DA3) with gradual change of electrons acceptors (from weakest to strongest) in the same solvent showed that the organic structures with strongest electron acceptors had faster decay of the fluorescence and hence a stronger intramolecular charge transfer. Moreover, the comparison of the two structures showed that the dendrimers molecules generally showed stronger intramolecular charge transfer. Also the experimental results showed that when the solvent effect ( interactions dipole – dipole) on spectral properties of molecules by reducing the energy of the excited levels (red shift Stoke) of the fluorescence spectra of molecules and then the structures with the strongest electron acceptors had faster decay of the fluorescence

Ce rapport de thèse présente une étude de la dynamique transitoire de molécules de taille réduite en solution induite par photoexcitation. Celle-ci est réalisée au travers d'une expérience de mesure de diffusion de rayons X aux grands angles (TR-WAXS). La partie principale de ce rapport s'articule autour de la dertermination des structures transitoires avec l'utilisation de plusieurs schémas structurelles d'optimisation. Certains résultats obtenus par TR-WAXS sont complétés avec ceux obtenus par spectroscopie optique résolue dans le temps (TOAS) ainsi que par spectroscopie d'émission de rayons X de ces molécules (TR-XES) permettant une plus complète description de ces systèmes.Ce travail est composé de 4 differents projets lesquelles peuvent être regroupé en 2 groupes selon le type de réaction declenchées par photoexcitation: dissociation et transfert d'électron. Le premier groupe comprends la molécule triatomique de mercure, halogénure métallique et le triiodide où nous étudions la dynamique de recombinaison des photofragments libres. Cette partie démontre la possibilité de développer de nouvelles connaissances concernant la dynamique de ces modèles, comme par exemple, la découverte de la recombinaison par canaux ou bien par l'apparition de réactions intermédiaires en moins de 100 ps, correspondant à la longueur de l'impulsion de rayons X générée par le synchrotron, mise en évidence en utilisant la technique de découpage temporelle. Le deuxième groupe de systèmes est composé de complexes de métaux de transition, très pertinents pour les applications liées à la récolte de lumière, [Fe(NHC)2]2+ et Ru=Co, où le spin et les structures dynamiques sont étudiées. Ces projets prolongent de précedents travaux à une famille de systèmes plus complexes avec un numéro atomique bas (première colonne de transition metallique) et fait la lumière sur de nouveaux composés en photochimie. / This thesis investigates photoinduced transient dynamics of small molecules in solution with the use of time-resolved wide angle x-ray scattering (TR-WAXS). The core of the thesis is the precise determination of transient structures with the use of various structural optimization schemes. Some of the TR-WAXS results are supplemented with transient absorption optical spectroscopy (TOAS) and time-resolved x-ray emission spectroscopy (TR-XES) measurements allowing for a more complete description of the systems.par The work includes four different projects which can be grouped into two classes according to the type of reaction appearing upon photoexcitation: dissociation and electron transfer. The first class includes the simple triatomic molecules mercury halides and triiodide where we study the recombination dynamics of the free photofragments. This part of the thesis demonstrates the possibility to gain new knowledge about the dynamics of these model systems such as new recombination channels and reaction intermediates appearing in less than 100 ps, the xray pulse length from the synchrotron, by the use of time-slicing technique. The second class of systems includes transition metal complexes that are highly relevant for light harvesting applications, [Fe(NHC)2]2+ and Ru=Co, where the spin and structural dynamics are investigated. These projects extend previous work to a family of more complex systems constituted of relatively low-Z atoms (first row transition metals) and bring new insights into the photochemistry of the compounds.

La possibilité de déterminer précisément et de quantifier la composition des milieux biologiques est un défi pour l'imagerie médicale qui permettrait de diagnostiquer certaines maladies ou de mieux étudier les processus physiologiques. La tomographie optique diffuse (DOT) est une technique d'imagerie attrayante qui permet de façon non invasive, non-ionisante et potentiellement avec une grande spécificité de sonder les milieux en profondeur en utilisant la lumière dans le proche infrarouge et de reconstruire en trois dimensions leur composition. Pour obtenir les caractéristiques des chromophores endogènes (oxy- et désoxy-hémoglobine) enfouis dans un milieu très diffusant, une instrumentation optique dans le domaine temporel et multi-longueurs d'onde combinée à un algorithme de reconstruction en trois dimensions a été développée. Des mesures expérimentales ont été menées en géométrie de réflectance en éclairant avec un laser picoseconde un milieu perturbé (contenant une hétérogénéité) et en récupérant, pour plusieurs longueurs d'onde et multi-positions, la lumière rétrodiffusée via deux fibres optiques connectées à deux détecteurs dédiés et couplés à un système de comptage de photon unique. Le traitement des données de ces mesures résolues en temps a été réalisé en supposant que la propagation de la lumière est gouvernée par l'approximation de la diffusion et en utilisant une méthode basée sur la transformée de Mellin-Laplace. Des simulations et des expériences sur une gamme de milieux imitant les milieux biologiques ont démontré que cette technique médicale a le potentiel pour donner des images médicales quantitatives. Nous avons montré que l'on peut déterminer correctement la composition d'objets à 10 mm de profondeur absorbant faiblement. Pour de plus fortes profondeurs et des absorptions plus élevées, la valeur du coefficient d'absorption ou de la concentration est sous-estimée. En outre, l'imagerie multimodale apporte des améliorations dans la précision de la quantification en profondeur et donc peut être une bonne opportunité pour les futures applications cliniques de la DOT. / In medical imaging, the ability to accurately retrieve and quantify the composition of turbid media is challenging and would enable to diagnose some diseases or to better study physiological processes. Diffuse optical tomography (DOT) is an attractive medical imaging technique which permits to probe in depth using near-infrared light and to reconstruct in three dimensions the composition of biological tissues non-invasively, non-ionizing and with potentially high specificity. To obtain endogenous chromophore (oxy- and desoxy-hemoglobin) features in the depth of a highly scattering medium, a multiwavelength time domain optical setup combined to a three-dimensional reconstruction algorithm was developed. Experimental measurements were conducted in reflectance geometry by illuminating a perturbed medium (with a heterogeneity) with a picosecond laser and by collecting, for several wavelengths and multi-positions, the backscattered light via two fibers connected to two dedicated detectors and coupled to a time-correlated single photon counting system. The data processing of these time-resolved measurements and those of a known reference medium was performed by supposing that the propagation of light is governed by the diffusion approximation and using a method based on Mellin-Laplace transform. Numerical and phantom experiments on series of objects similar to biological media demonstrate that this technique has the potential to give quantitative medical images. We have highlighted a correct quantification for the less absorbing objects at 10 mm depth while underestimation results at deeper depths and higher absorptions. Furthermore, the multimodal imaging brings improvements in quantification in depth and thus it can be a good opportunity to DOT for its future clinical applications.

No description available.

In the field of physical chemistry, the relatively recently developed technique of velocity-map imaging has allowed chemical dynamics to be explored with a greater depth than could be previously achieved using other methods. Capturing the scattering image associated with the products resulting from fragmentation of a molecule allows the dissociative pathways and energy landscape to be investigated. In the study of particle physics, the neutron has become an irreplaceable spectroscopic tool due to the unique nature of the interaction with certain materials. Neutron spectroscopy is a non-destructive imaging technique that allows a number of properties to be discerned, including chemical identification, strain tensor measurements and the identification of beneath the sample surface using radiography and tomography. In both of these areas, as well as a multitude of other disciplines, a flight tube is used to separate particles, distinguishing them based upon their mass in the former case and their energy in the latter. The experiments can be vastly enhanced by the ability to record both the position and arrival time of the particle of interest. This thesis describes several new developments made in instrumentation for experiments involving time-of-flight particle imaging. The first development described is the construction of a new velocity-map imaging instrument that utilises electron ionisation to perform both steps of molecular fragmentation and ionisation. Data from CO2 is presented as an example of the ability of the instrument, and a preliminary analysis of the images is performed. The second presented project is the design of a time-resolved and position-resolved detector developed for ion imaging experiments. The hardware, software and firmware are described and presented alongside data from a variety of the experiments showcasing the breadth of investigations that are possible using the sensor. Finally, the modifications made to the detector to allow time-resolved neutron imaging are detailed, with an in-depth description of the various proof-of-concept experiments carried out as part of the development process.

Ce travail de thèse s’inscrit dans l’étude des matériaux moléculaires photo-commutables. L’objectif est le contrôle réversible et persistant des propriétés physiques de la matière par application d’un stimulus externe, tel qu’une excitation optique. Le travail de thèse est constitué de deux parties principales. La première partie traite de la photo-isomérisation des ligands NO dans les complexes de ruthénium di-nitrosyle. Une démarche expérimentale systématique a été effectuée sur trois complexes. La spectroscopie infrarouge et la diffraction des rayons-X ont été utilisées conjointement à basse température. De multiples isomères de liaison photo-induits (PLI) ont été détectés. Il a été montré qu’il est possible de photo-commuter chacun des deux ligands nitrosyles indépendamment. De plus, une double photo-isomérisation a été caractérisée. La deuxième partie présente le développement et la mise en œuvre d’une expérience de diffraction des rayons-X résolue en temps en laboratoire. Le dispositif est un diffractomètre prototype avec un détecteur à pixel hybride XPAD, permettant des mesures à l’échelle de la milliseconde. Un système pompe-sonde est couplé au diffractomètre permettant une excitation périodique de l’échantillon et une caractérisation in-situ de la photo-commutation. Cette expérience a été validée par la mesure de l’état métastable MSII du complexe nitrosyle Na2[Fe(CN)5NO].2H2O, d’une durée de vie de 12 ms à 150 K / This work is part of the study of photo-switchable molecular materials. The objective is the reversible and persistent control of the physical properties of matter by the application of an external stimulus, such as optical excitation. The thesis work consists of two main parts. The first part deals with the photoisomerization of NO ligands in ruthenium di-nitrosyl complexes. A systematic experimental approach was performed on three complexes. Infrared spectroscopy and X-ray diffraction were used together at low temperature. Multiple photoinduced linkage isomers (PLI) were detected. It has been shown that it is possible to photo-switch each of the two nitrosyl ligands independently. In addition, a double photoisomerization was characterized. The second part presents the development and implementation of an in-house time-resolved X-ray diffraction experiment. The device is a prototype diffractometer with a XPAD hybrid pixel detector, allowing measurements on a millisecond time-scale. A pump-probe system is coupled to the diffractometer allowing periodic excitation of the sample and an in-situ characterization of the photo-switching. This experiment was validated by measuring the metastable state MSII of the nitrosyl complex Na2[Fe(CN)5NO].2H2O, with a lifetime of 12 ms at 150 K

Dans ce travail, nous avons étudié le mécanisme d'émission de la partie optiquement active du complexe luciférine-luciférase. Ce système bioluminescent est largement utilisé dans un très grand nombre d'approches bioanalytiques. Ce phénomène naturel résulte en l'émission de lumière visible (vert-jaune-rouge) à partir du photoproduit : l’oxyluciférine. Une des hypothèses couramment admise pour expliquer le mécanisme d’émission de l’oxyluciféreine fait intervenir un équilibre complexe entre six espèces chimiques, mais le détail exact du mécanisme reste à élucider. Les principales conclusions présentées ici repose sur l'identification des six formes chimiques de l’oxyluciférine impliquées dans le mécanisme d'émission de fluorescence et la caractérisation d’un point de vu dynamique du transfert de proton à l’état excité. Ces résultats ont été obtenus par l'étude des propriétés optiques de différents analogues structuraux de l’oxyluciférine dans un tampon aqueux. Différent techniques de spectroscopie (état stable et résolue en temps) et des approches chimiométriques ont été appliquées pour étudier ce mécanisme d'émission. En outre, les propriétés photophysiques de l’oxyluciférine en complexe avec l'enzyme luciférase (Luciola cruciata) ont été étudiées également en milieu aqueux. En parallèle, les dérivés présentant des propriétés d’émission sensibles à l’environnement ont été utilisés pour visualiser l'interaction entre biomolécules. En particulier, nous avons démontré que l’oxyluciférine peut être utilisée pour cartographier le pH intracellulaire à l'aide de la microscopie de fluorescence dans des cellules vivantes. Avec l'aide d'un autre dérivé de l'oxyluciférine nous avons été en mesure de caractériser l'interaction entre une protéine du VIH-1 et des séquences d'oligonucléotide au moyen de mesures ratiométriques. Enfin, nous avons développé une approche basée sur le marquage de résidus cystéine pour suivre, in vitro, l'interaction protéine-protéine. / In this work, we investigated the emission mechanism of the optically active part of the firefly luciferin-luciferase complex. This bioluminescent system is widely used in bioanalytical assay. This amazing natural phenomenon results in the emission of visible light (yellow-green-red) from the photoproduct Oxyluciferin. This color tuning mechanism involves six chemical species, but their active involvement in the excited state proton transfer (ESPT) mechanism was poorly understood so far. One of the main finding presented here relies on the identification of six chemical forms of Oxyluciferin involved in the color tuning fluorescence emission mechanism. This result was obtained by studying the optical properties of different structural analogues of firefly Oxyluciferin in aqueous buffer. Different spectroscopic (steady state and time-resolved) and chemometric approaches have been applied to reveal the emission mechanism. In addition, the photophysical properties of Oxyluciferin in complex with the Luciferase enzyme Luciola cruciata have been studied in aqueous buffer as well. In parallel, derivatives displaying environment sensitive emission were used to monitor biomolecular interactions. In particular, we demonstrated that Oxyluciferin can be employed to map intracellular pH by using fluorescence microscopy within living cells. With the help of another Oxyluciferin derivative we were able to monitor the interaction between a HIV-1 protein and different oligonucleotide sequences by means of ratiometric measurements. Finally we develop an approach based on cysteine labeling to monitor in vitro protein-protein interaction.

A compreensão dos caminhos de desativação não radiativa em polímeros conjugados é fundamental para o uso desses materiais em dispositivos luminescentes, células fotovoltaicas e sensores. Nesta tese, os processos não radiativos em filmes automontados de polímero luminescente foram investigados via análise da supressão de intensidade de fotoluminescência, utilizando a técnica de fluorescência estacionária, e através da medida do tempo de vida do estado excitado com fluorescência resolvida no tempo em arranjo de contagem de fótons. A camada emissiva constituída por poli(p-fenileno vinileno) (PPV) obtido da conversão térmica do precursor poli(cloreto de xilideno tetraidrotiofeno) (PTHT) apresentou tempos de vida entre 150 e 250 ps, e fraca dependência com o comprimento de onda de detecção da emissão. Na presença de uma monocamada contendo azocorante Vermelho do Congo (CR) ou ftalocianina tetrassulfonada de níquel (NiTsPc) atuando como receptor de energia foi observada supressão da fluorescência e encurtamento do tempo de vida (50-100ps). À medida que se aumenta a distância entre as camadas de doador e receptor via interposição de bicamadas de espaçador, a dinâmica do sistema tende à dinâmica de decaimento do estado excitado do PPV na ausência de receptor. A dependência da eficiência com a distância 1/r2 ou 1/r3 apontam para transferência de energia ressonante (RET) típica da interação plano-plano ou plano-camada. Os processos não radiativos foram também estudados em função da diluição de segmentos PPV dispersos em matriz de polieletrólito poli(cloreto de dodecildimetilamônio) PDAC. A diluição dos segmentos conjugados aumenta a eficiência quântica de emissão, torna o mecanismo de desativação mais lento (500 ps) e diminui a dependência do tempo de vida com o comprimento de onda de detecção, devido à menor eficiência dos mecanismos de desativação não radiativa. Na presença de uma camada receptora, a eficiência de transferência de energia é maior para amostras com segmentos mais diluídos indicando que esse mecanismo compete com os processos internos à própria camada emissiva. O tempo de vida aumenta em filmes com mais baixo grau de conjugação e a dependência com o comprimento de onda de emissão é mais forte. Em resumo, a técnica de automontagem se mostrou adequada para obter filmes com propriedades fotofísicas controláveis no nível molecular, que puderam ser estudadas com espectroscopias de fluorescência. / Understanding the pathway of excitation in conjugated polymers is important for using these materials as active layer in devices, photovoltaic cells and sensors. In this thesis, non-radiative processes in layer-by-layer (LbL) films were investigated with measurements of fluorescence quenching in steady-state fluorescence spectroscopy and of the singlet excited state lifetime using time-resolved fluorescence spectroscopy in a single photon counting (SPC) apparatus. The emissive layer of poly(p-phenylene vinylene) (PPV) obtained via thermal conversion of the soluble precursor poly(xylylidene tetrahydrothiophenium) chloride (PTHT) showed lifetime 150-250 ps and weak dependence on the emission wavelength. In the presence of an acceptor layer of either Congo Red (CR) or nickel tetrasulfonated phthalocyanine (NiTsPc) the PL intensity was quenched and the lifetime was shorter (50 -100 ps). Upon increasing the distance between the energy donor (PPV) and acceptor layers by interposing bilayers of inert polyelectrolytes the dynamics tended to the PPV dynamics. The distance-dependence of the energy transfer efficiency (1/r2 or 1/r3) points to a resonant process (RET) with a plane-to-plane or plane-to-slab interaction. The non-radiative processes were also studied for varying dilutions of the PPV segments in a matrix of the polyelectrolyte poly(dodecyl methylamonium chloride) (PDAC). This dilution increased the quantum efficiency and led to longer lifetimes (500 ps) owing to a less effective non-radiative energy transfer. When an energy acceptor layer is present, energy transfer is more efficient for the most diluted systems, indicating that such transfer competes with internal processes in the emissive layer. The lifetime increased in films with low degree of conjugation, for which there was a stronger dependence on the emission wavelength. It is concluded that the LbL methodology was suitable to obtain photophysical film properties that could be controlled at the molecular level, and investigated with varied fluorescence spectroscopies.

Submitted by Geandra Rodrigues (geandrar@gmail.com) on 2017-12-20T13:20:23Z No. of bitstreams: 1 israelnovaesdemoura.pdf: 1420476 bytes, checksum: 6634c39e49385490d2f7b019fb451af1 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-12-21T11:59:42Z (GMT) No. of bitstreams: 1 israelnovaesdemoura.pdf: 1420476 bytes, checksum: 6634c39e49385490d2f7b019fb451af1 (MD5) / Made available in DSpace on 2017-12-21T11:59:42Z (GMT). No. of bitstreams: 1 israelnovaesdemoura.pdf: 1420476 bytes, checksum: 6634c39e49385490d2f7b019fb451af1 (MD5) Previous issue date: 2017-08-04 / O objetivo deste trabalho foi avaliar a caracterização e eficácia de diferentes técnicas da espectroscopia de fluorescência na detecção de adulterações em formulações de Whey Protein concentrado (WPC) em pó, a partir de sua mistura com substâncias de diferentes origens. Foram estudados dois diferentes lotes de WPC provenientes do mesmo fornecedor. Adulterações foram realizadas a partir da adição individual de Cafeína (Tratamento 1), Creatina (Tratamento 2) e Lactose (Tratamento 3) a 30% (m/m) em WPC e submetidas às análises de espectroscopia de fluorescência estacionária e resolvida no tempo, utilizando-se os comprimentos de onda de excitação de 275 e 335 nm. Quando excitadas a 275 nm, as amostras apresentaram pico de emissão a 335 nm aproximadamente, com uma banda de emissão em torno de 380 nm, característica apenas na amostra contendo cafeína, enquanto lactose e creatina não induziram alterações no espectro do WPC. Quando excitadas a 335 nm, as amostras apresentam picos de emissão com máximo em 425 e 470 nm, sem diferenciação por simples observação do espectro. Análise da distância Euclidiana evidenciou que, quando excitadas a 275 nm, os espectros completos dos tratamentos 2 e 3 foram semelhantes ao Controle 1 enquanto o Tratamento 1 foi diferente. Já na excitação a 335 nm todos os espectros foram semelhantes. Análise de Componentes Principais (PCA) confirmou a diferenciação do Tratamento 1 a 275 nm mas foi inconclusiva com a excitação de 335 nm, porém determinou pontos de interesse para estudos das derivadas dos espectros. Com as derivadas, foi possível a diferenciação entre os Tratamentos 2 e 3 nos dois comprimentos de onda, com enfoque em comprimentos de ondas específicos que podem ser decisivos na diferenciação das adulterações. Em relação a espectroscopia resolvida no tempo, o Tratamento 1 demonstrou valores da intensidade média do tempo de vida de emissão superior aos tratamentos 2 e 3 nos dois comprimentos de onda de excitação empregados. A adulteração com cafeína foi realizada na amostra Controle 2 e foi observado resultado semelhante quando comparada ao Controle 1. De maneira geral, a aplicação das técnicas de espectroscopia de fluorescência estacionária e resolvida no tempo possibilitou a caracterização das amostras utilizadas no estudo. Além disso, possibilitou a observação de diferenças entre as amostras controles e aquelas adulteradas, especialmente a adicionada de cafeína e excitada no comprimento de onda 275 nm, com ajuda de ferramentas matemáticas. Os resultados aqui obtidos corroboram com o fato de que as técnicas empregadas podem ser importantes na detecção de fraudes em produtos lácteos. / The objective of this work was to evaluate the characterization and efficacy of different fluorescence spectroscopy techniques in the detection of adulterations in formulations of Whey Protein Concentrate (WPC) powder, from its mixture with substances of different origins. Two different batches of WPC from the same supplier were studied. Adulterations were performed from the individual addition of Caffeine (Treatment 1), Creatine (Treatment 2) and Lactose (Treatment 3) at 30% (w / w) in WPC and subjected to stationary fluorescence spectroscopy and time-resolved fluorescence, using the excitation wavelengths of 275 and 335 nm. When excited at 275 nm, the samples showed an emission peak at approximately 335 nm, with an emission band around 380 nm, characteristic only in the sample containing caffeine, while lactose and creatine did not induce alterations in the WPC spectrum. When excited at 335 nm, the samples showed peak emission at 425 and 470 nm, without differentiation by simple observation of the spectrum. Euclidean distance analysis showed that, when excited at 275 nm, the complete spectra of treatments 2 and 3 were similar to Control 1 while Treatment 1 was different. Regarding the excitation at 335 nm, all spectra were similar. Principal Component Analysis (PCA) confirmed the differentiation of Treatment 1 at 275 nm but it was inconclusive with 335 nm excitation, however, it determined points of interest for spectra derivative studies. With the derivatives, it was possible to differentiate between Treatments 2 and 3 in the two wavelengths, focusing on specific wavelengths that can be decisive in the differentiation of adulterations. In relation to time resolved fluorescence spectroscopy, Treatment 1 demonstrated values of the mean emission lifetime higher over Treatments 2 and 3 at the two excitation wavelengths employed. The caffeine adulteration was performed in the Control 2 sample and a similar result was observed when compared to Control 1. In general, the application of stationary and time resolved fluorescence spectroscopy techniques allowed the characterization of the samples used in the study. In addition, it made possible the observation of differences between the control and adulterated samples, especially the one with caffeine added and excited at the wavelength 275 nm, with the help of mathematical tools. The results obtained here corroborate the fact that the techniques employed may be important in the detection of fraud in dairy products.