Matrix merging arrangements for the study protein dynamics by time-resolved step-scan Fourier transform infrared spectroscopy and multivariate curve resolution

In this paper, we propose the construction of merging arrangements for combining the information from various runs as a powerful approach to improve the resolution. The bacteriorhodopsin (bR) photocycle has been chosen in this study as an example dealing with the protein dynamics monitored by means of time-resolved step-scan FT-IR spectroscopy. The possibilities of matrix merging are evaluated and results are compared with those from the analysis of individual and augmented matrices. As a conclusion, this strategy provides excellent results for the analysis of this type of time-resolved FT-IR data.     

Time-resolved flow-flash FT-IR difference spectroscopy: the kinetics of CO photodissociation from myoglobin revisited

Fourier-transform infrared (FT-IR) difference spectroscopy has been proven to be a significant tool in biospectroscopy. In particular, the step-scan technique monitors structural and electronic changes at time resolutions down to a few nanoseconds retaining the multiplex advantage of FT-IR. For the elucidation of the functional mechanisms of proteins, this technique is currently limited to repetitive systems undergoing a rapid photocycle. To overcome this obstacle, we developed a flow-flash experiment in a miniaturised flow channel which was integrated into a step-scan FT-IR spectroscopic setup. As a proof of principle, we studied the rebinding reaction of CO to myoglobin after photodissociation. The use of microfluidics reduced the sample consumption drastically such that a typical step-scan experiment takes only a few 10 ml of a millimolar sample solution, making this method particularly interesting for the investigation of biological samples that are only available in small quantities. Moreover, the flow cell provides the unique opportunity to assess the reaction mechanism of proteins that cycle slowly or react irreversibly. We infer that this novel approach will help in the elucidation of molecular reactions as complex as those of vectorial ion transfer in membrane proteins. The potential application to the oxygen splitting reaction of cytochrome c oxidase is discussed. An erratum to this article can be found at     

Ordering and mobility of ferroelectric liquid crystal dimer as studied by FT-IR spectroscopy with 2D-IR correlation analysis

Both a conservative rapid-scan FT-IR technique and a novel step-scan FT-IR technique with 2D correlation analysis were used to study the orientation and the mobility of a ferroelectric liquid crystal dimer during switching under an electric field. The detailed mutual arrangements of different molecular segments (mesogen, poly(methylene) chain, polysiloxane chain) in a smectic C* phase were derived from the static spectra. It was shown that the long mesogen axis, the average poly(methylene) and the average polysiloxane chain axes do not coincide with each other. The hindered rotation of the carbonyl group is confirmed. Time-resolved FT-IR technique was used to follow the segmental motion with a time resolution of 5 μs. The temperature and electric field strength dependencies of the mobility of these segments are discussed. 2D correlation analysis of time-resolved data reveals small differences in the behavior of the carbonyl and the benzoic rings in the mesogen moieties, that can be explained as differences in the orientation distribution functions of these moieties.     

Observations of different triplet conformations in time-resolved infrared spectra of alkyl phenylglyoxylates

Two different conformations of the triplet state of alkyl phenylglyoxylates were observed by means of time-resolved step-scan FT-IR spectroscopy. The amplitude of the peak corresponding to the sterically hindered conformation decreases as the size of the alkyl group increases. Both conformations exhibit similar reactivity in intermolecular hydrogen abstraction, but only one of them undergoes Norrish Type II photoelimination.     

Set-up for time-resolved step-scan FTIR spectroscopy of noncyclic reactions

We have established a novel technique, which allows the application of time-resolved step-scan FTIR difference spectroscopy on noncyclic reactions. Cyclic reactions are ideally suited for the step-scan technique. However, it is difficult to apply the step-scan technique to noncyclic reactions, because the investigated process has to be repeated at about 1000 sampling positions of the interferogram. Consequently, to investigate noncyclic systems the sample has to be renewed at every sampling position. In the presented novel approach the IR-beam and the excitation laser-beam are focused to a very small diameter of 200 μm. Thereby, only a small segment of the sample, which has an overall diameter of 15 mm, is excited and probed. By moving the sample, which is mounted on an x-y-stage, to different nonexcited segments the reaction can be repeated until a complete interferogram data set is recorded. In so far as the typically used flow cells are concerned their optical pathlength is too large to perform difference spectroscopy. We use 4 μm thin films to depress the water background absorption of biological samples. As test, the well known photo-cyclic reactions of bacteriorhodopsin are measured. No systematic errors appear in the difference spectra. Because of intensity loss by the IR-microscope the signal-to-noise ratio is about 5 times less as compared to conventional step-scan measurements. For the first time, the technique is then applied to a noncyclic reaction, the photolysis of caged ATP. The successful performance with 10 μs time-resolution now opens the door for many new applications of step-scan FTIR measurements to noncyclic reactions.     

Step-scan near-infrared Fourier-transform Raman spectroscopy with 100 picosecond laser pulses

Time-resolved Fourier-transform Raman scattering with picosecond excitation is reported for the first time. The resonance Raman spectrum of 9,10-diphenylanthracene in its excited single state was obtained by Raman excitation at 1064 nm with 100 ps pulses, following photoexcitation at 355 nm. The implementation and characterization of time-resolved FT-Raman spectroscopy with a step-scan interferometer is discussed. FT-Raman spectra generated with continuous and mode-locked beams in the continuous-scan mode of the interferometer are compared with step-scan FT-Raman spectra generated with 2 kHz pulses.     

Fourier-transform infrared study of the switching process in a ferroelectric liquid crystalline polymer

The implementation of the step-scan technique on our FT-IR spectrometer enabled us to follow the electric-field induced reorientation dynamics of different molecular segments of a ferroelectric liquid crystalline polymer on a sub-millisecond time scale. It was detected that not only the mesogen but also the spacer and at least part of the backbone take part in the reorientation process.     

Segmental orientation and mobility of ferroelectric liquid crystal polymers

IR spectroscopy was used to study the orientation and mobility of different molecular segments in a side chain ferroelectric liquid crystalline polymer (FLCP) in the book-shelf geometry. It was directly shown that the tilt angles for the mesogenic units and the spacers are different. The data obtained allowed us to construct a detailed model of segmental orientation in the SC phase for this FLCP. This model is consistent with the 'zigzag' model for tilted smectic phases. The rotational bias of carbonyl bonds is also confirmed and a possible orientation function for the carbonyl group is discussed. Time-resolved step-scan FTIR spectroscopy enabled us to follow the intra- and inter-molecular response of the FLCP to an external electric field with a time resolution of 5 mus. It was detected that mesogenic moiety, spacer and backbone take part in the reorientation process. The time responses of different molecular segments are similar on the time scale of a few hundred microseconds.     

FT-IR/UATR and FT-IR transmission quantitative analysis of PBT/PC blends

Polymer blends are produced to achieve properties that pure polymers are unable to. The qualitative analysis of these blends is not always satisfactory to point out failures in the final product and, therefore, the determination of their content might be required. The aim of this study is to develop quantitative methodologies using Fourier transform infrared spectroscopy (FT-IR), reflection mode with the universal attenuated total reflectance accessory (UATR) and transmission techniques, for the determination of the content of each polymer present in the polymer blend poly (butylene terephthalate) (PBT) and polycarbonate (PC). FT-IR/UATR presented better results using the relative band (A₁₇₇₂/A₁₇₁₆). FT-IR/UATR presented error of 2.30%, which is a little higher than the accuracy limit of the spectrometer (≤2%), but still adequate for industrial applications.     

Structural Properties and Stress in ZnO Films Obtained from a Nanocolloidal Sol

This paper deals with stress analysis of ZnO films, via substrate curvature measurements. The films were prepared from a nanocolloidal sol by dip-coating and annealed from 150 to 400°C under pure or ozone-enriched oxygen. The chemical film evolution is followed by FT-IR spectroscopy, and the structural properties by X-ray diffraction and optical absorption spectroscopy. It is shown that the films annealed under pure oxygen are in a compressive stress state due to a stuffing effect during grain growth, whereas the films annealed under ozone-enriched oxygen are in a tensile stress state due to pore collapsing. This astonishing difference has been attributed to the skeleton, thinner in the latter than in the former films. Indeed, the acidifying action of ozone on alcohols is believed to be responsible for slowing down the grain growth during annealing.     

Determination of oxidation products in transformer oils using FT-IR spectroscopy

Transformer oils are subject to a lot of control checks in the process of their exploitation, because the presence of oxidized products causes damages in the electrical equipment. In the process of exploitation of transformer oils anti-oxidation additives get exhausted and products of oxidation are formed. This paper describes the development method for study of oxidation transformer oils by Fourier transform infrared (FT-IR) spectroscopy. The aim of this paper is to determine the main products of oxidation of transformer oils and to increase the sensitivity of FT-IR analysis by the extraction method. The method which we suggest to register all oxidized products is of great interest. The problem is the detection limit of routine apparatus. For this reason we have developed an extraction method, which can identify precisely all oxidized products qualitatively and quantitatively in the region 1800-1650 cm⁻¹. Thus a manifold increase of the FT-IR analysis is achieved, combined with increased sensitivity, precision and the possibilities for registering micro-amounts otherwise beyond the routine technique. FT-IR spectroscopy is a powerful method with great practical significance for investigation of carbonyl- and carboxylic degradation products from transformer oils.     

Evaluation of the renal calculi compositions

Thermal analysis and infrared (IR) spectroscopy are modern physical–chemical methods suitable for the investigation of the kidney stones composition. The applications of these methods in our work were anticipated by performing the standard thermal analysis and standard infrared spectra on pure compounds: oxalates, phosphate, carbonate, and uric acid. This work reveals a logical algorithm for correlating the experimental data regarding urolithiasis types and compositions by thermal analysis methods, Fourier Transform Infrared (FT-IR) spectroscopy, and second derivative FT-IR spectra. The limits and performance of each analysis method have been highlighted, and by correlating the results of both methods we have obtained comprehensive information for the identification and/or determination of the main components as well as of organic and/or inert impurities in calculi composition. The data regarding the urinary calculi composition are important for the clinical guideline in the prophilaxy and methaphylaxy of urolithiasis.     

一种面向多组分定量的通用谱图解析方法

在一个多组分系统中各组分的定量分析是非常重要的。现代光谱如拉曼光谱(RS)、傅里叶变换红外(FT-IR)光谱、紫外-可见(UV-Vis)光谱、核磁共振(NMR)、质谱(MS)等,可通过获取丰富峰信号的谱图对样品进行各角度的详尽描述。然而,由于谱图的复杂性及其解析工作的繁重,使得仅通过样品谱图来同时量化混合物中的每个组分成为很具挑战性的工作。在这项研究中,我们首次介绍了一个名为定量主成分分析(q PCA)的可靠策略,快速计算混合物中每个成分的比例,而不需要任何手动解谱。通过使用纯组分的谱图作为参考,多组分系统的谱图可以通过PCA自动分辨并解析,然后就可以使用我们的计算方法来计算每个组分的比例。计算机建模实验和RS、FT-IR、UV-Vis、NMR、MS实验都证明了这一策略胜任多组分系统的快速定量工作。     

Photoacoustic infrared spectroscopy of Syncrude post-extraction oil sand

Rapid- and step-scan photoacoustic (PA) infrared spectra of three fractions of a Syncrude post-extraction oil sand were analyzed in detail in this work. The rapid-scan spectra showed that the samples were comprised primarily of kaolinite, quartz, silica, siderite, and residual hydrocarbons, and that the proportions of these constituents were different for each fraction. Depth profiling of the three post-extraction oil sands was accomplished using both rapid- and step-scan PA infrared spectroscopy. The results confirmed that kaolinite is more abundant in the near-surface region, whereas quartz and hydrocarbons are concentrated at greater depths. The modulation frequency dependence of the PA intensities for all three fractions was consistent with a model in which the samples are thermally thick; in other words, the thermal diffusion length (roughly equal to the sampling depth) was less than the particle sizes of all three samples. The results of this study are consistent with published reports on the PA infrared spectra of fine tailings generated during bitumen extraction and the spectroscopic and thermophysical characterization of clay soils and an appropriate model clay.     

Liquid exclusion adsorption chromatography, a new technique for isocratic separation of nonionic surfactants. III. Two-dimensional separation of fatty alcohol ethoxylates

In this paper, a novel procedure for qualitative and quantitative analysis of the two-dimensional data obtained from GC-MS is investigated to determine chemical components of essential oils in Cortex Cinnamomi from four different producing areas. A new method named iterative optimization procedure (IOP) specially used to resolve embedded peaks is also developed. With the help of IOP and other chemometric techniques, such as heuristic evolving latent projections, evolving factor analysis, sub-window factor analysis and orthogonal projection resolution, and etc., the detection of the purity of chromatographic peaks can be first addressed, and then the overlapping peaks are resolved into the pure chromatogram and mass spectrum of each component. The similarity searches in the MS database are finally conducted to qualitatively determine the chemical components. The results obtained showed that the accuracy of qualitative and quantitative analysis could be greatly enhanced by chemometric resolution methods. The chemometric resolution techniques upon the two-dimensional data can be quite promising tools for the analysis of the complex samples like traditional Chinese medicine.     

Monitoring of molecular transformations in acid-base reactions by evolving factor analysis of Fourier transform infrared spectral data

An evolving factor analysis procedure with concentration constraints (gradient concentration window) was applied to the analysis of data sets of aqueous Fourier transform infrared (FT-IR) spectra of carboxylic acids (acetic, malonic and succinic acids) collected in experiments with varying pH. Besides the calculation of the number of acid-base systems, this procedure allowed the calculation of the FT-IR spectra of the acid-base species present in equilibrium as well as the corresponding pK(a) values.     

电化学原位步进扫描时间分辨显微镜FTIR反射光谱

结合步进扫描傅立叶变换红外光谱仪和红外显微镜，建立了电化学原位步进扫描时间分辨显微镜FTIR反射光谱.采用微电极和利用纳米结构表面的异常红外效应，显著提高表面吸附物种的红外检测灵敏度和红外电极表面对极化电位的响应速度，使广泛应用的外反射型薄层红外电解池的时间常数(τ=R1Cd)从使用常规盘电极的约40 ms降低到0.65 ms.研究了酸性介质中CO在Pt电极上的吸附过程，谱图的时间分辨率达50 μs.     

X-ray absorption spectroscopic investigation of partially reduced cobalt species in Co-MCM-41 catalysts during synthesis of single-wall carbon nanotubes

Chemometric tools were employed to analyze the in-situ dynamic X-ray absorption spectroscopy data to probe the state of Co-MCM-41 catalysts during reduction in pure hydrogen and under single-wall carbon nanotube synthesis reaction conditions. The use of the progressive correlation analysis established the sequence in which changes in the spectral features near the Co K edge occurred, and the evolving factor analysis provided evidence for the formation of an intermediate Co(1+) ionic species during reduction of the Co-MCM-41 catalyst in pure hydrogen up to 720 degrees C. This intermediate species preserves the tetrahedral environment in the silica framework and is resistant to complete reduction to the metal in H(2). While the Co(2+) species is resistant to reduction in pure CO, the intermediate Co(1+) species is more reactive in CO most likely forming cobalt carbonyl-like compounds with high mobility in the MCM-41. These mobile species are the precursors of the metallic clusters growing carbon nanotubes. Controlling the rates of each step of this two-stage reduction process is key to controlling the size of the metallic Co clusters formed in Co-MCM-41 catalysts.     

Spectroscopic characterisation and identification of ecdysteroids using high-performance liquid chromatography combined with on-line UV--diode array, FT-infrared and 1H-nuclear magnetic resonance spectroscopy and time of flight mass spectrometry

A prototype multiply hyphenated reversed-phase HPLC system has been applied to the analysis of a mixture of pure ecdysteroids and an ecdysteroid-containing plant extract. Characterisation was achieved via a combination of diode array UV, ¹H NMR, FT-IR spectroscopy and time of flight (TOF) mass spectrometry. This combination of spectrometers allowed the collection of UV, ¹H NMR, IR and mass spectra for a mixture of pure standards enabling almost complete structural characterisation to be performed. The technique was then applied to a partially purified plant extract in which 20-hydroxyecdysone and polypodine B were identified despite incomplete chromatographic resolution and the presence of co-chromatographing interferents. The experimental difficulties in the use of such a systems for these analytes are described.     

Infrared study of chlorophenols and products of their photodegradation

The presents study of the 3-chlorophenol, 2,4-dichlorophenol, 2,3,4,5-tetrachlorophenol, pentachlorophenol and products of their photodegradation using FT-IR spectroscopy and GC/MS. Spectra of pure chlorophenols with the spectra of their solutions after photodegradation were compared. FT-IR spectra of pure chlorophenols investigated in the region of 3700–3000 cm⁻¹ show that in particular cases the position and shape of bands corresponding to stretching vibration of hydroxyl groups are different. In all cases, the differences between spectra of pure chlorophenols and irradiated solutions were observed. It was confirmed that different distribution of types of hydrogen bonds appearing in particular chlorophenols has strong influence on the process of irradiation reaction and final products.