Quantification of individual fatty acids in bovine milk by infrared spectroscopy and chemometrics: understanding predictions of highly collinear reference variables.

Predicting individual fatty acids (FA) in bovine milk from Fourier transform infrared (FT-IR) measurements is desirable. However, such predictions may rely on covariance structures among individual FA and total fat content. These covariance structures may change with factors such as breed and feed, among others. The aim of this study was to estimate how spectral variation associated with total fat content and breed contributes to predictions of individual FA. This study comprised 890 bovine milk samples from 2 breeds (455 Holstein and 435 Jersey). Holstein samples were collected from 20 Danish dairy herds from October to December 2009; Jersey samples were collected from 22 Danish dairy herds from February to April 2010. All samples were from conventional herds and taken while cows were housed. Moreover, in a spiking experiment, FA (C14:0, C16:0, and C18:1 cis-9) were added (spiked) to a background of commercial skim milk to determine whether signals specific to those individual FA could be obtained from the FT-IR measurements. This study demonstrated that variation associated with total fat content and breed was responsible for successful FT-IR-based predictions of FA in the raw milk samples. This was confirmed in the spiking experiment, which showed that signals specific to individual FA could not be identified in FT-IR measurements when several FA were present in the same mixture. Hence, predicted concentrations of individual FA in milk rely on covariance structures with total fat content rather than absorption bands directly associated with individual FA. If covariance structures between FA and total fat used to calibrate partial least squares (PLS) models are not conserved in future samples, these samples will show incorrect and biased FA predictions. This was demonstrated by using samples of one breed to calibrate and samples of the other breed to validate PLS models for individual FA. The 2 breeds had different covariance structures between individual FA and total fat content. The results showed that the validation samples yielded biased predictions. This may limit the usefulness of FT-IR-based predictions of individual FA in milk recording as indirect covariance structures in the calibration set must be valid for future samples. Otherwise, future samples will show incorrect predictions.

Flaxseed dietary fibers suppress postprandial lipemia and appetite sensation in young men.

BACKGROUND AND AIM: Dietary fibers (DF) are linked to a reduced risk of life-style diseases, which relate to their physiological effects in the gastrointestinal tract. The aim was to examine whether flaxseed DF-enriched meals suppress postprandial lipemia and reduce appetite. METHODS AND RESULTS: Four different iso-caloric meals were tested in 18 young men in a double-blind randomized crossover design. Test meals were served after an overnight fast. DF content and source were: control (C): 1.4 g/MJ; whole flaxseed (WF): 2.4 g/MJ from whole flaxseeds; low-mucilage dose (LM): 2.4 g/MJ from flaxseed DF; high-mucilage dose (HM): 3.4 g/MJ from flaxseed DF. During the 7 h test day, subjective appetite sensation was assessed using visual analogue scales and appetite-regulating hormones, and lipemia and glycemia were measured, after which ad libitum energy intake was recorded. There was a significant time × meal effect on triacylglycerols (TG) (p = 0.02) and an 18% smaller area under the curve (AUC) for TG after meal HM compared to meal C was observed (p < 0.01). AUC for insulin was smaller after both LM and HM meals compared to C and WF meals. Higher mean ratings of satiety (p < 0.01) and fullness (p = 0.03) was seen following the HM meal compared to meal C. AUC for ghrelin, CCK and GLP-1 and ad libitum energy intake did not differ between meals, but ghrelin response exhibited a different response pattern after the mucilage-containing meals. CONCLUSION: These findings suggest that flaxseed DF may suppress postprandial lipemia and appetite although subsequent energy intake was not affected.

Exploring abiotic stress on asynchronous protein metabolism in single kernels of wheat studied by NMR spectroscopy and chemometrics.

Extreme climate events are being recognized as important factors in the effects on crop growth and yield. Increased climatic variability leads to more frequent extreme conditions which may result in crops being exposed to more than one extreme event within a growing season. The aim of this study was to examine the implications of different drought treatments on the protein fractions in grains of winter wheat using (1)H nuclear magnetic resonance spectroscopy followed by chemometric analysis. Triticum aestivum L. cv. Vinjett was studied in a semi-field experiment and subjected to drought episodes either at terminal spikelet, during grain-filling or at both stages. Principal component trajectories of the total protein content and the protein fractions of flour as well as the (1)H NMR spectra of single wheat kernels, wheat flour, and wheat methanol extracts were analysed to elucidate the metabolic development during grain-filling. The results from both the (1)H NMR spectra of methanol extracts and the (1)H HR-MAS NMR of single kernels showed that a single drought event during the generative stage had as strong an influence on protein metabolism as two consecutive events of drought. By contrast, a drought event at the vegetative growth stage had little effect on the parameters investigated. For the first time, (1)H HR-MAS NMR spectra of grains taken during grain-filling were analysed by an advanced multiway model. In addition to the results from the chemical protein analysis and the (1)H HR-MAS NMR spectra of single kernels indicating that protein metabolism is influenced by multiple drought events, the (1)H NMR spectra of the methanol extracts of flour from mature grains revealed that the amount of fumaric acid is particularly sensitive to water deficits.

Differentiation of perirenal and omental fat quality of suckling lambs according to the rearing system from Fourier transforms mid-infrared spectra using partial least squares and artificial neural networks analysis.

Fourier transform mid-infrared (FT-IR) spectroscopy was evaluated as a tool to discriminate between carcasses of suckling lambs according to the rearing system. Fat samples (39 perirenal and 67 omental) were collected from carcasses of lambs from up to three sheep dairy farms, reared on either ewes milk (EM) or milk replacer (MR). Fatty acid composition of the samples from each fat deposit was first analyzed and, when discriminant-partial least squares regression (PLS) was applied, a perfect discrimination between rearing systems could be established. Additionally, FT-IR spectra of fat samples were obtained and discriminant-PLS and artificial neural network (ANN) based analysis were applied to data sets, the latter using principal component analysis (PCA) or support vector machines (SVM) as processing procedure. Perirenal fat samples were perfectly discriminated from their FT-IR spectra. However, analysis of omental fat showed misclassification rates of 9-13%, with the ANN approach showing a higher discrimination power.

Quantitative analysis of NMR spectra with chemometrics.

The number of applications of chemometrics to series of NMR spectra is rapidly increasing due to an emerging interest for quantitative NMR spectroscopy e.g. in the pharmaceutical and food industries. This paper gives an analysis of advantages and limitations of applying the two most common chemometric procedures, Principal Component Analysis (PCA) and Multivariate Curve Resolution (MCR), to a designed set of 231 simple alcohol mixture (propanol, butanol and pentanol) (1)H 400 MHz spectra. The study clearly demonstrates that the major advantage of chemometrics is the visualisation of larger data structures which adds a new exploratory dimension to NMR research. While robustness and powerful data visualisation and exploration are the main qualities of the PCA method, the study demonstrates that the bilinear MCR method is an even more powerful method for resolving pure component NMR spectra from mixtures when certain conditions are met.

Application of the NMR-MOUSE to food emulsions.

The application of the NMR-MObile Universal Surface Explorer (NMR-MOUSE) to study food systems is evaluated using oil-in-water emulsions, and the results are compared to those obtained using a conventional low-field NMR (LF-NMR) instrument. The NMR-MOUSE is a small and portable LF-NMR system with a one-sided magnet layout that is used to replace the conventional magnet and probe on a LF-NMR instrument. The high magnetic field gradients associated with the one-sided MOUSE magnet result in NMR signal decays being dominated by molecular diffusion effects, which makes it possible to discriminate between the NMR signals from oil and water. Different data acquisition parameters as well as different approaches to the analysis of the NMR data from a range of oil-in-water emulsions are evaluated, and it is demonstrated how the concentration of oil and water can be determined from the NMR-MOUSE signals. From these model systems it is concluded that the NMR-MOUSE has good potential for the quantitative analysis of intact food products.

The diluted aqueous solvation of carbohydrates as inferred from molecular dynamics simulations and NMR spectroscopy.

The purpose of this paper is to review our understanding of the dilute hydration (aqueous solvation) behaviour of disaccharide compounds. To this end we discuss and scrutinize the results that have been obtained for the three model disaccharides: maltose, sucrose and trehalose from experimental NMR studies and from theoretical molecular dynamics studies in explicit aqueous solutions. The focus is on the description of molecular hydration features that will influence macroscopic entities such as diffusion and relaxation: residence times of hydration waters, hydration numbers and hydration densities. The principles of molecular dynamics simulation are briefly outlined while a detailed presentation is given of the key features that characterise hydration: the solvation of the glycosidic linkage, the radial hydration of the solute, the water density anisotropy around the solute, the residential behaviour of water molecules in the periphery of the solute, and rotational and translational diffusion coefficients. With respect to the use of NMR in characterising the structure and dynamics of the hydration, the hydrodynamic theory of rotational and translational diffusion of biomolecules as well as the use of pulse field gradient spin echo experiments are briefly presented. The NMR-defined rotational diffusion coefficients (D(r)) and the experimentally determined translational diffusion (D(t)) coefficients are reported for 4% (w/w) solutions of sucrose, trehalose and maltose. These results are compared with theoretical data obtained from molecular dynamics simulations of sucrose, trehalose and maltose under identical conditions (concentration, temperature, etc.). With our present level of knowledge we can propose that although carbohydrates share a number of hydration characteristics, evidence is accumulating in support of the notion that it is not the amount or overall hydration but rather the detailed individual carbohydrate-water interaction that is likely to determine carbohydrate structure and functionality.

Internal motions and hydration of sucrose in a diluted water solution.

This paper represents a synthesis of our most recent work on the hydration, internal and overall dynamics of sucrose in a diluted water solution. The studies were carried out as a total ensemble of 1.2 nanosecond condensed phase molecular dynamics trajectories. In this study the focus is on a 500 ps trajectory starting with the solute in the crystalline conformation. The presence of water was found to significantly alter the accessible conformational space of the solute. All potential intra-molecular hydrogen bonds were found to be exchanged to surrounding water molecules and the simulations suggest that the sucrose conformation is stabilized by the dynamic presence of two interring bridging water molecules: O-2g...Ow...O-3f and O-2g...Ow...O-lf. The overall shift in conformation of the solute induced by the presence of water was found to improve the theoretical models of experimental traits. It is demonstrated that the hydration structure and the internal and overall motions of sucrose compare extremely well with NMR data such as glycosidic heteronuclear coupling constants and the molecular tumbling time, with X-ray data of two partially hydrated sucrose structures in a protein complex and with translational diffusion coefficients and hydration numbers established from experimental studies.

The mean hydration of carbohydrates as studied by normalized two-dimensional radial pair distributions.

The hydration of carbohydrates plays a key role in many biological processes. Molecular dynamics simulations provide an effective tool for investigating the hydration of complex solutes such as carbohydrates. In this article we devise an algorithm for the calculation of two-dimensional radial pair distributions describing the probability of finding a water molecule in a site defined by two reference atoms. The normalized 2D radial pair distribution is proposed as an effective tool for investigating and comparing localized or ordered water sites around flexible molecules such as carbohydrates when analyzing molecular dynamics simulations and the utility of 2D radial pair distributions is demonstrated using sucrose as an example. In this relatively simple structure, 2D radial pair distributions were able to characterize and quantify the importance of two unique interresidue hydration sites in which a water molecule is forming a bridge between the glycopyranosyl and fructofuranosyl residues. The approach is proposed to be a valuable tool for comparing and understanding the hydration of flexible biomolecules such as carbohydrates.

Travelling on the potential energy surfaces of carbohydrates: comparative application of an exhaustive systematic conformational search with an heuristic search.

The calculated ensembles found by a heuristic conformational search algorithm, CICADA, for three small carbohydrates, ethyl beta-lactoside, methyl alpha-D-galactoside, and methyl beta-D-galactoside, are evaluated in terms of their ability to reproduce time-averaged optical rotation and NMR data. A unique dynamic model for methyl beta-D-galactoside has been obtained by fitting experimental NOESY volumes to the theoretical ones elaborated from the CICADA ensemble internuclear distances with the model-free formalism. In the case of ethyl beta-lactoside, the CICADA ensemble is compared to that of an exhaustive systematic grid-search method. The CICADA algorithm proved to be a very efficient method to find most of the important minima on even very complex potential energy surfaces, and the spectral quality of the CICADA ensemble was found to be of equal quality, if not superior, to that of the exhaustive systematic grid-search method. The CICADA algorithm has several advantages over other conformational search algorithms: (1) It has polynomial dependence of dimensions on computer time in contrast to the grid search, which has exponential dependence, (2) the conformations found are free of artificial harmonic constraint potentials, (3) it passes all barriers amongst families of conformations on conformational hypersurface but spends almost all its time in the essential highly populated areas, (4) the inherent properties of the algorithm make rigorous minimization criteria superfluous and provide good convergence behavior, and (5) as an important spin-off, it provides low-energy interconversion pathways that can, amongst others, be used for estimating adiabatic rotational barriers.

Evaluation of quality changes in walnut kernels (Juglans regia L.) by Vis/NIR spectroscopy.

Storage of walnut kernels in light and at room temperature, as is common practice, is detrimental to their sensory quality and shelf life. This study demonstrates that Vis/NIR spectroscopy, in combination with multivariate data analysis (chemometrics), is a most capable rapid method for monitoring the overall quality deterioration of walnut kernels. Spectral predictions of the sensory attributes nutty and rancid tastes by partial least-squares regression (PLSR) resulted in correlations (r(2)) of 0.77 and 0.86, respectively, whereas with PLSR prediction of the chemical parameter hexanal content a correlation (r(2)) of 0.72 was obtained. The study further establishes that storage in light results in pronounced oxidative changes, especially in walnuts stored at 21 degrees C, whereas dark storage at 5 degrees C results in walnuts without any trace of rancid taste during 25 weeks of storage at accelerated storage conditions (50% oxygen).

Presence and dehydration of ikaite, calcium carbonate hexahydrate, in frozen shrimp shell.

Ikaite, calcium carbonate hexahydrate, has by means of X-ray diffraction analyses of frozen samples been identified as the mineral component of the white spots formed in the shell of frozen shrimp during storage. When the shrimp thaw and the shell material is dried and kept at room temperature, ikaite rapidly transforms into a mixture of anhydrous calcium carbonate forms. X-ray diffraction analyses and Raman spectra of synthetic ikaite as well as the dehydration product confirm the assignments, and the rate constant for dehydration is approximately 7 x 10(-)(4) s(-)(1) at ambient temperature. Differential scanning calorimetry showed that dehydration of synthetic ikaite is an entropy-driven, athermal process and confirms that a single first-order reaction is rate-determining. Ikaite is found to be stable in aqueous solution at temperatures below 5 degrees C and in the shell of frozen shrimps but decomposes on thawing to form anhydrous calcium carbonates.

Conformations of disaccharides by empirical force field calculations. Part V: Conformational maps of beta-gentiobiose in an optimized consistent force field.

A recently optimized set of potential energy functions is used to investigate the conformational flexibility of the beta-(1-->6) glycosidic linkage in beta-gentiobiose. Relaxed Ramachandran maps in vacuo are presented in the torsional angles phi and omega, with torsional angle psi allowed to relax freely, as are all other internal degrees of freedom. The study reveals two almost iso-energetic low energy domains in (phi, psi, omega) space, and cross-sections in the low-energy domains at omega = -60 degrees and omega = 60 degrees show that more than 60% and 70% respectively of the area of the conformational maps are accessible within 40 kJ mol-1. The molecular structure in the crystal, including the exoanomeric effect, is well reproduced. The structure belongs to the potential energy well which includes the global potential energy minimum in vacuo. The most profound structural difference between the crystal structure and the calculated global minimum in vacuo is the 20 degrees deviation of the psi torsional angle (24 degrees from perfect trans). It occurs in the most flexible glucosidic degree of freedom, psi, and is caused by optimization of the hydrogen bonding network and not by the exoanomeric effect.

Prediction of water-holding capacity and composition of porcine meat by comparative spectroscopy.

Four spectroscopic instruments, a fibre optical probe (FOP), a visual (VIS) and near infrared (NIR) reflectance spectrophotometer, a reflectance spectrofluorometer and a low-field (1)H nuclear magnetic resonance (LF-NMR) instrument were used to perform measurements on two muscles (longissimus dorsi and semitendinosous) from 39 pigs, 18 of which were carriers of the Halothane gene. Water-holding capacity (drip loss and filter paper wetness) and chemical composition (intramuscular fat and water) of the muscle samples were determined for spectroscopic calibration. Prediction models were established by partial least squares regression to evaluate the potential of using the spectroscopic techniques in an on-line slaughterhouse system. VIS data gave good prediction models, indicating that current industrial colour systems can be advanced into more specific meat evaluation systems by including the entire visible spectral range. The FOP and fluorescence measurements were less successful, and suffered from sampling problems since they measure only a small area. The best regression models were obtained from LF-NMR data for all reference quality measures and yielded a correlation coefficient of 0.75 with drip loss. LF-NMR proved able to distinguish between the two muscles and the results for their longitudinal relaxation times, T(21), were proportional to their average myofibrillar cross-sectional areas reported in the literature.

Warmed-over flavour in porcine meat - a combined spectroscopic, sensory and chemometric study.

The effectiveness of applying rapid spectral techniques in the prediction of meat quality in relation to pre-slaughter stress and warmed-over flavour (WOF) was investigated. The effect of pre-slaughter stress on the development of WOF is a relatively new area of interest in WOF research. The present study investigated the relationship between pre-slaughter stress and WOF characteristics (after 0 to 5 days' storage) in porcine meat as evaluated by chemical, sensory and spectroscopic methods. Sensory evaluation, visual reflectance spectroscopy (VIS) and low-field (1)H NMR (LF-NMR) proved efficient in describing the different stress groups and the storage period, whereas fluorescence spectroscopy and the TBARS test were only able to follow WOF during storage. Near Infrared Reflectance (NIR) and Raman scatter showed a very weak relationship to pre-slaughter stress and levels of WOF during storage. Good correlations (up to r=0.93) of sensory terms were achieved with VIS and fluorescence spectroscopy and LF-NMR.

icoshift: A versatile tool for the rapid alignment of 1D NMR spectra.

The increasing scientific and industrial interest towards metabonomics takes advantage from the high qualitative and quantitative information level of nuclear magnetic resonance (NMR) spectroscopy. However, several chemical and physical factors can affect the absolute and the relative position of an NMR signal and it is not always possible or desirable to eliminate these effects a priori. To remove misalignment of NMR signals a posteriori, several algorithms have been proposed in the literature. The icoshift program presented here is an open source and highly efficient program designed for solving signal alignment problems in metabonomic NMR data analysis. The icoshift algorithm is based on correlation shifting of spectral intervals and employs an FFT engine that aligns all spectra simultaneously. The algorithm is demonstrated to be faster than similar methods found in the literature making full-resolution alignment of large datasets feasible and thus avoiding down-sampling steps such as binning. The algorithm uses missing values as a filling alternative in order to avoid spectral artifacts at the segment boundaries. The algorithm is made open source and the Matlab code including documentation can be downloaded from www.models.life.ku.dk.

A molecular builder for carbohydrates: application to polysaccharides and complex carbohydrates.

A new procedure (POLYS) for producing three-dimensional structures of polysaccharides and complex carbohydrates is described. This employs a builder concept combining a database of monosaccharide structures with a database containing information on populations of independent neighboring glycosidic linkages in disaccharide fragments. The computer program is written in C, and it can cope with both the complexity and the diversity of carbohydrates and the unique topological features arising from multiple branching. A simple ASCII syntax was developed for describing the primary structures in accordance with IUPAC nomenclature. The translation of the primary structure is made through the combined use of a lexical analyzer and a command interpreter. In this way the program can be considered as a compiler of primary structures of carbohydrates. However, it also generates secondary and tertiary structures in the form of Cartesian coordinates in formats used by most molecular mechanics programs and packages. In our laboratory POLYS was exhaustively tested on standard homopolysaccharide systems such as cellulose and mannan and found to work very well. We now report the ease of use and the efficiency of the molecular builder in applications to more complex carbohydrate systems. These include the structural exploration of a pentaantennary oligosaccharide having 135 residues, the complex family of pectic polysaccharides including the organization and distribution of side chains (arabinan, arabinogalactan, and galactan) on the rhamnogalacturonan backbone.

Modeling polysaccharides: present status and challenges.

The most recent tools that have been developed for modeling the three-dimensional features of polysaccharides and carbohydrate polymers are presented. The presentation starts with a description of the conformations of the monosaccharides, and of the flexible rings such as in the case of five-membered rings, and a thorough description of the conformational space that is available for a disaccharide unit, either in vacuo or in an aqueous phase. The extension to the modeling of the parent polysaccharides is addressed, based on the assumption that owing to the size and relative rigidity of the intervening monosaccharides units, the rotations at a particular linkage can be, under some conditions, considered as independent of nearest neighbor interactions. Appropriate modeling techniques are described that can provide insights into the dimensions of the chain in a solution which is best described as a random coil accompanied by the occurrence of local "helical" regions. With the help of such descriptors such as helical parameters, the ordered state of polysaccharide strands can be readily characterized. The generation of double or triple helices can be then attempted in order to explore the occurrence of such multi-stranded arrangements that may be energetically stable. The final step in the determination of the structure of polysaccharides in the ordered state, is the investigation of the interactions of different helices. This may lead to either the best arrangement(s) between two polymeric chains, or to the prediction of the dimensions, and the symmetry of a three-dimensional lattice. Some of the tools which have been developed should allow automatic scarches for meaningful correlations between structures and functions, through exploratory data analysis. Structure-function or structure-property correlation could be then used to model changes arising from structural alterations. This would open the field of polysaccharide engineering.