Molecular-level insight into hot-melt loading and drug release from mesoporous silica carriers.

Drug amorphisation by loading to inorganic mesoporous carriers represents an emerging area of improving the dissolution rate and bioavailability of poorly water-soluble active pharmaceutical ingredients (APIs). In this work, for the first time, a molecular-level insight into the process of API loading to mesoporous SiO2 (silica) carriers by the hot-melt impregnation method and its subsequent release during dissolution was obtained using ATR-FTIR spectroscopic imaging. A physical mixture of ibuprofen crystals and mesoporous silica particles was heated and the dynamics of melt loading into the silica pore structure was directly observed in situ by ATR-FTIR spectroscopic imaging. The loss of crystallinity, the redistribution of the API in the silica pore network and the subsequent stabilisation of the amorphous form upon cooling were proven. The API was involved in two different kinds of molecular-level interactions: API dimers in the amorphous bulk, and individual API molecules adsorbed on the silica surface. The melt-loaded silica carriers were comprehensively characterised by DSC, SEM and dissolution tests, which proved dissolution rate enhancement due to amorphisation of the API. Drug release form the hot-melt loaded mesoporous silica carriers was observed in real time and the conditions leading to local re-crystallisation of super-saturated solution of the API were identified.

Protein hydration in living cells probed by Fourier transform infrared (FT-IR) spectroscopic imaging.

An application of disrelation mapping to Fourier transform infrared (FT-IR) spectroscopic imaging datasets is provided to reveal different H-bonded water species within a mammalian cell. 2D correlation analysis revealed a disrelation peak at (3420 cm-1, 3220 cm-1), showing the existence of a specific water band at around 3220 cm-1 whose variation of absorbance did not follow the trend of water molecules with a well-coordinated H-bonding network. Disrelation maps constructed with disrelation intensities between (3420 cm-1, 3220 cm-1) and (3420 cm-1, 1540 cm-1) reveal that the disruption of the water network occurs around the interfacial regions between the cell and media, indicating the disintegration of the H-bonding network of bulk water due to the entrapment of water by the protein inside the cell. This hydration effect also becomes apparent around the area adjacent to the cellular nucleus, reflecting the fact that protein synthesis mainly occurs in this region. These results clearly show the presence of different molecular states of water inside living cells, which are not readily identified by conventional analysis methods.

Analyses of trace amounts of dyes with a new enhanced sensitivity FTIR spectroscopic technique: MU-ATR (metal underlayer ATR spectroscopy).

The identification of organic dyes is a challenging task in all the fields such as the forensic and conservation sciences, especially in cases where the amount of sample is extremely small. In this paper we propose a new enhanced FTIR method (MU-ATR metal underlayer ATR spectroscopy), which we believe is the first of its kind, for the analysis of a few ng of dyes. With this method, dyed fiber micro-extracts can be analyzed using a commercial FTIR microscope with a fixed incident angle, obtaining the same separation between the different classes of dyes investigated as we obtained analyzing pure dyes in transmission mode. Moreover, the new enhancement method has been validated on a real sample dated back to the 1893, showing how it can be promising for the analysis of trace amounts of organic substances in artistic samples such as dyes in paintings or textiles, varnishes and organic residues on archaeological objects.

Nanopatterning and tuning of optical taper antenna apex for tip-enhanced Raman scattering performance.

This paper focuses on finding optimal electrochemical conditions from linear sweep voltammetry analysis for preparing highly reproducible tip-enhanced Raman scattering (TERS) conical gold tips with dc-pulsed voltage etching. Special attention is given to the reproducibility of tip apex shapes with different etchant mixtures. We show that the fractional Brownian motion model enables a mathematical description of the decaying current kinetics during the whole etching process up to the cutoff event. Further progress in preparation of highly reproducible smooth and sharp tip apexes is related to the effect of an additive, such as isopropanol, to aqueous acids. A finite-difference time-domain method based near-field analysis provides evidence that TERS performance depends critically on tip orientation relative to a highly focused laser beam. A TERS based criterion for recognizing gold tips able to couple/decouple optical near- and far-fields is proposed.

Generation of chemical movies: FT-IR spectroscopic imaging of segmented flows.

We have previously demonstrated that FT-IR spectroscopic imaging can be used as a powerful, label-free detection method for studying laminar flows. However, to date, the speed of image acquisition has been too slow for the efficient detection of moving droplets within segmented flow systems. In this paper, we demonstrate the extraction of fast FT-IR images with acquisition times of 50 ms. This approach allows efficient interrogation of segmented flow systems where aqueous droplets move at a speed of 2.5 mm/s. Consecutive FT-IR images separated by 120 ms intervals allow the generation of chemical movies at eight frames per second. The technique has been applied to the study of microfluidic systems containing moving droplets of water in oil and droplets of protein solution in oil. The presented work demonstrates the feasibility of the use of FT-IR imaging to study dynamic systems with subsecond temporal resolution.

pH-sensitive polymer hydrogels derived from morpholine to prevent the crystallization of ibuprofen.

Various pharmaceutical formulations of ibuprofen are available including oral tablets, suspensions, suppositories and transdermal gels and creams. Aqueous ibuprofen solubility is dependent on pH increasing solubility when increasing pH. The very low aqueous solubility of ibuprofen can lead to a segregation of the drug in to microdomains and possible crystallization which will affect the release profile even leading to possible damage of the mucous membrane of the stomach. Therefore, homopolymers and copolymers of N-ethylmorpholine methacrylamide (EMA) and N, N-dimethylacrylamide (DMA) hydrogels have been prepared to be applied as matrices for ibuprofen release as a method for minimizing these issues. The hydrogels were loaded with ibuprofen and the release over time was tested at 37°C and at different pH values: 2, 5 and 7.4. Fourier-transform infrared spectroscopy with attenuated total reflection (FTIR-ATR) was performed in order to identify the stretching vibrations of the carbonyl group for each component. Dissolution of the ibuprofen from the formulations at different pHs was studied using the ATR-FTIR spectroscopic imaging. The results showed that for DMA hydrogels, most of ibuprofen was released as non crystalline ibuprofen at pH 7.4 but it was not able to prevent crystallization at pH 2 and 5. In contrast, the EMA hydrogels were able to prevent crystallization of the ibuprofen at all pHs. Finally, it was demonstrated that there is a minimum concentration at which the polymer becomes ineffective.

ATR-FTIR spectroscopy and spectroscopic imaging of solvent and permeant diffusion across model membranes.

The uptake and diffusion of solvents across polymer membranes is important in controlled drug delivery, effects on drug uptake into, for example, infusion bags and containers, as well as transport across protective clothing. Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) spectroscopy has been used to monitor the effects of different solvents on the diffusion of a model compound, 4-cyanophenol (CNP) across silicone membrane and on the equilibrium concentration of CNP obtained in the membrane following diffusion. ATR-FTIR spectroscopic imaging of membrane diffusion was used to gain an understanding of when the boundary conditions applied to Fick's second law, used to model the diffusion of permeants across the silicone membrane do not hold. The imaging experiments indicated that when the solvent was not taken up appreciably into the membrane, the presence of discrete solvent pools between the ATR crystal and the silicone membrane can affect the diffusion profile of the permeant. This effect is more significant if the permeant has a high solubility in the solvent. In contrast, solvents that are taken up into the membrane to a greater extent, or those where the solubility of the permeant in the vehicle is relatively low, were found to show a good fit to the diffusion model. As such these systems allow the ATR-FTIR spectroscopic approach to give mechanistic insight into how the particular solvents enhance permeation. The solubility of CNP in the solvent and the uptake of the solvent into the membrane were found to be important influences on the equilibrium concentration of the permeant obtained in the membrane following diffusion. In general, solvents which were taken up to a significant extent into the membrane and which caused the membrane to swell increased the diffusion coefficient of the permeant in the membrane though other factors such as solvent viscosity may also be important.

Local examination of skin diffusion using FTIR spectroscopic imaging and multivariate target factor analysis.

In the context of trans-dermal drug delivery it is very important to have mechanistic insight into the barrier function of the skin's stratum corneum and the diffusion mechanisms of topically applied drugs. Currently spectroscopic imaging techniques are evolving which enable a spatial examination of various types of samples in a dynamic way. ATR-FTIR imaging opens up the possibility to monitor spatial diffusion profiles across the stratum corneum of a skin sample. Multivariate data analyses methods based on factor analysis are able to provide insight into the large amount of spectroscopically complex and highly overlapping signals generated. Multivariate target factor analysis was used for spectral resolution and local diffusion profiles with time through stratum corneum. A model drug, 4-cyanophenol in polyethylene glycol 600 and water was studied. Results indicate that the average diffusion profiles between spatially different locations show similar profiles despite the heterogeneous nature of the biological sample and the challenging experimental set-up.

Application of principal component analysis to the thermal characterization of silanized nanoparticles obtained at supercritical carbon dioxide conditions.

Samples resulting from reaction of TiO(2) with octyltriethoxysilane, developed using a supercritical carbon dioxide procedure, have been studied by infrared and Raman spectroscopies and thermogravimetric analysis. Different reaction conditions have been applied to the preparation of samples in order to study the influence of experimental factors on the sample properties. Vibrational techniques have first been used to verify the presence of silanized structures on the surface of TiO(2) through the detection of specific bands characteristic of the Si-O-Si cross-linking. Thermogravimetric profiles consisting of weight loss values as a function of temperature have been analyzed by principal component analysis to extract information about the characteristics of the linkage between silane and TiO(2) as well as the thermal stability of the prepared materials. The mathematical treatment of data has provided conclusions on the properties of the samples and analogies and differences with respect to the commercial material.

Applications of ATR-FTIR spectroscopic imaging to biomedical samples.

FTIR spectroscopic imaging in ATR (Attenuated Total Reflection) mode is a powerful tool for studying biomedical samples. This paper summarises recent advances in the applications of ATR-FTIR imaging to dissolution of pharmaceutical formulations and drug release. The use of two different ATR accessories to obtain chemical images of formulations in contact with water as a function of time is demonstrated. The innovative use of the diamond ATR accessory allowed in situ imaging of tablet compaction and dissolution. ATR-FTIR imaging was also applied to obtain images of the surface of skin and the spatial distribution of protein and lipid rich domains was obtained. Chemical images of cross-section of rabbit aorta were obtained using a diamond ATR accessory and the possibility of in situ imaging of arterial samples in contact with aqueous solution was demonstrated for the first time. This experiment opens an opportunity to image arterial samples in contact with solutions containing drug molecules. This approach may help in understanding the mechanisms of treatment of atherosclerosis.

Fourier transform infrared imaging of human hair with a high spatial resolution without the use of a synchrotron.

The cross-section of a human hair has been imaged for the first time using the micro attenuated total reflection (ATR) Fourier transformed infrared (FT-IR) method in combination with a focal plane array (FPA) detector. A rigorous approach was applied to determine the spatial resolution, namely, measuring the distance over which the band absorbance changes from 95 to 5% of the maximum absorbance when passing through a sharp interface. The measured value for IR transmission was approximately 16 microm, while the value obtained using ATR imaging was approximately 5 microm. The enhanced spatial resolution achieved by this method allows the medulla of the hair (approximately 8 microm in diameter) to be imaged clearly without the need for a synchrotron source. The spatial resolution of transmission and ATR imaging is compared, and advantages of ATR imaging are discussed.

Effects of particle size on near-wall depletion in mono-dispersed colloidal suspensions.

In this work we investigate the change in particle concentration near a solid boundary for colloidal dispersions in pressure driven flow, commonly referred to as wall depletion. In particular we determine the effect of Peclet number on the strength and spatial extent of the depleted layer. The change in concentration near the solid boundary is measured with attenuated total reflection infrared (ATR-IR) spectroscopy described previously (P.J.A. Hartman Kok et al., J. Rheol. 46 (2002) 481). The method is capable of measuring the concentration of particles at distances ranging from 0.2 to 1.0 mum from the boundary. The suspensions investigated consisted of mono-dispersed polystyrene particles in water. Particles of four different sizes were used, with radius, a, of 30, 54, 105, and 197 nm. (The ratio H/a was in the range 2500-17,000 with H being the height of the flow cell.) This enabled us to measure the wall depletion effect over a wide range of Peclet numbers, ranging from 0.01 to 45. We found that wall depletion was not significant for Peclet numbers smaller than unity. Estimates of the wall slip layer thickness obtained from rheological experiments were consistent with the results obtained by ATR-IR spectroscopy.

Spectroscopic imaging of arteries and atherosclerotic plaques.

Fourier transform infrared (FTIR) spectroscopic imaging using a focal plane array detector has been used to study atherosclerotic arteries with a spatial resolution of 3-4 microm, i.e., at a level that is comparable with cellular dimensions. Such high spatial resolution is made possible using a micro-attenuated total reflection (ATR) germanium objective with a high refractive index and therefore high numerical aperture. This micro-ATR approach has enabled small structures within the vessel wall to be imaged for the first time by FTIR. Structures observed include the elastic lamellae of the tunica media and a heterogeneous distribution of small clusters of cholesterol esters within an atherosclerotic lesion, which may correspond to foam cells. A macro-ATR imaging method was also applied, which involves the use of a diamond macro-ATR accessory. This study of atherosclerosis is presented as an illustrative example of the wider potential of these ATR imaging approaches for cardiovascular medicine and biomedical applications.

New opportunities in micro- and macro-attenuated total reflection infrared spectroscopic imaging: spatial resolution and sampling versatility.

New opportunities exist to obtain chemical images using attenuated total reflection infrared (ATR-IR) spectroscopy. This paper shows the feasibility of obtaining FT-IR images with a spatial resolution of at least 3-4 microm using a Ge ATR objective coupled with an infrared microscope. The improved spatial resolution compared to FT-IR images obtained by the transmission method is due to the high refractive index of the ATR crystal, which gives a high numerical aperture and hence, a higher spatial resolution. FT-IR imaging with a conventional diamond ATR accessory has been investigated. This is the first time that FT-IR imaging is reported using such a versatile accessory based on a diamond ATR crystal. These results showed that a spatial resolution up to 13 microm can be achieved without the use of infrared microscope objectives. One advantage of the diamond element is that it allows pressure to be applied and hence, good contact to be obtained over the whole field of view.

Confocal Raman microscopy of supercritical fluid dyeing of polymers.

Supercritical CO2 was used as a medium to introduce azo-dyes into polymers (PMMA and PET). Confocal Raman microscopy has been applied using dry metallurgical and oil objectives to measure the concentration of dye as a function of distance from the surface to the core of the polymer samples. A significant gradient of dye concentration normal to the surface, down to the depth of ca. 100 microm, has been observed. In addition, cross-sections of cut dyed polymer samples have been analysed using a conventional mapping approach. The comparison between confocal depth and conventional mapping approaches has demonstrated that the use of the oil objective can provide reliable qualitative information on dye distribution in these systems. These confocal Raman experiments demonstrate the potential of this method in applications to polymeric materials processed with supercritical fluids.

Spectroscopy of polymer/drug formulations processed with supercritical fluids: in situ ATR-IR and Raman study of impregnation of ibuprofen into PVP.

In situ ATR (attenuated total reflectance)-IR spectroscopy has been used to study poly(vinylpyrrolidone) (PVP) films subjected to a solution of ibuprofen in supercritical CO2. The process of impregnation of ibuprofen into PVP has been monitored in situ. It has been shown that the supercritical fluid impregnation process results in ibuprofen being molecularly dispersed in a polymer matrix with ibuprofen molecules interacting with the C=O group of PVP. Raman spectra of ibuprofen impregnated into PVP from supercritical fluid solution have also been measured and compared with the Raman spectra of crystalline ibuprofen. ATR-IR spectroscopy has also revealed specific interactions between the C=O groups of PVP and CO2. Impregnation of ibuprofen into PVP makes the C=O groups of PVP less available for interactions with CO2. It has also been demonstrated that the presence of ibuprofen in PVP also affects sorption of water into PVP.

Membrane transport of hydrocortisone acetate from supersaturated solutions; the role of polymers.

Permeation of hydrocortisone acetate (HA) from supersaturated solutions was studied across a model silicone membrane. Supersaturated solutions were prepared using the cosolvent technique with propylene glycol and water (or aqueous polymer solutions) as the cosolvents. In the absence of the polymer, the flux of HA was similar at all degrees of saturation and was not significantly different from the value obtained for a saturated solution. Flux enhancement, as a result of supersaturation, was observed with all the polymers. The flux increased with increasing polymer concentration, reached a maximum and decreased at higher polymer percentages. The amount of polymer required for maximum enhancement differed for each polymer. The decrease of flux at high polymer concentrations is attributed to changes in microviscosity and a marginal increase in solubility. The infrared spectroscopic and differential scanning calorimetry data suggest that HA-polymer interactions occurred through hydrogen bonding thus explaining the proposed mechanism of the anti-nucleant properties of the polymers.