•A new method for effectively reconstructing diffusion-ordered spectroscopy.•It can give a good estimate of the number of chemical components.•It can resolve more chemical components than widely used multivariate methods.•It’s good at analyzing a mixture of mono- and poly-disperse species.2D diffusion-ordered NMR spectroscopy (DOSY) has been widely recognized as a powerful tool for analyzing mixtures and probing inter-molecular interactions in situ. But it is difficult to differentiate molecules with similar diffusion coefficients in presence of overlapped spectra. Its performance is susceptible to the number of chemical components, and usually gets worse when the number of components increases. Here, to alleviate the problem, numerical simultaneous inversion of Laplace transform (SILT) of many related variables is proposed for reconstructing DOSY spectrum (SILT-DOSY). The advantage of the proposed method in comparison to other methods is that it is capable of estimating the number of analytes more accurately and deriving corresponding component spectra, which in turn leads to the more reliable identification of the components.Download high-res image (62KB)Download full-size image

•13C NMR spectroscopy facilitates the assignment and deconvolution.•Sucrose, glucose and glutamate are the main cause of the discrimination.•The addition of caramel and monosodium glutamate was found.Soy sauce a well known seasoning all over the world, especially in Asia, is available in global market in a wide range of types based on its purpose and the processing methods. Its composition varies with respect to the fermentation processes and addition of additives, preservatives and flavor enhancers. A comprehensive 1H NMR based study regarding the metabonomic variations of soy sauce to differentiate among different types of soy sauce available on the global market has been limited due to the complexity of the mixture. In present study, 13C NMR spectroscopy coupled with multivariate statistical data analysis like principle component analysis (PCA), and orthogonal partial least square-discriminant analysis (OPLS-DA) was applied to investigate metabonomic variations among different types of soy sauce, namely super light, super dark, red cooking and mushroom soy sauce. The main additives in soy sauce like glutamate, sucrose and glucose were easily distinguished and quantified using 13C NMR spectroscopy which were otherwise difficult to be assigned and quantified due to serious signal overlaps in 1H NMR spectra. The significantly higher concentration of sucrose in dark, red cooking and mushroom flavored soy sauce can directly be linked to the addition of caramel in soy sauce. Similarly, significantly higher level of glutamate in super light as compared to super dark and mushroom flavored soy sauce may come from the addition of monosodium glutamate.The study highlights the potentiality of 13C NMR based metabonomics coupled with multivariate statistical data analysis in differentiating between the types of soy sauce on the basis of level of additives, raw materials and fermentation procedures.

The effects of temperature and NaCl on the micellization of CHAPS, a zwitterionic detergent widely used in membrane protein studies, have been investigated by NMR spectroscopy. We found that the two apparent critical micelle concentration (cmc) values of CHAPS decrease with the increase of temperature, as well as the NaCl concentration. The thermodynamic parameters derived from the temperature-dependent cmc values show that the micellization process is spontaneous and exothermic, and the van der Waals interaction is likely to be the main factor for the micellization of CHAPS. The micellar hydrodynamic radii remain almost the same in a range of 100−600 mM NaCl, indicating that the aggregate states of CHAPS are not sensitive to the change of the surrounding conditions. In addition, the dependence of nuclear Overhauser effect (NOE) intensities on temperatures further demonstrates the existence of the unique staggered micellar structure of CHAPS at a concentration above the apparent second cmc, which was suggested in our previous work. Our results provide a basis for optimizing CHAPS concentration in the solubilization or stabilization of membrane proteins under nondenaturing conditions and may be helpful to understand its interaction with proteins.

As an effective method for solvent suppression, WATERGATE is widely used in high resolution NMR spectroscopy. It is usually composed of a number of pulses separated by constant intervals. However, theoretical and experimental analyses indicate that narrower bandwidth and lower intensities around the secondary suppression points occur in the excitation profile of the composite WATERGATE. The excitation profile distortion is caused by the chemical shift evolution during the RF pulses. The higher the ratio of pulse duration to the inter-pulse delay is, the severer the profile distorts. Therefore, in high magnetic fields, the effect will be serious when WATERGATE is applied to some biological samples whose resonances distribute over a wide range. As can be seen obviously by applying WATERGATE to detect a RNA–protein mixture sample in an 800 MHz spectrometer, the resonances of the imino protons were partially suppressed by showing decreased intensities, though the intended secondary suppression points were set far away from them. In this article, we proposed an optimized WATERGATE that could effectively compensate the chemical shift evolution during the RF pulses, and relieve the excitation profile distortion. The optimized experiment will be a good way to retain the imino signal intensities when WATERGATE is applied to detect the RNA samples in high magnetic field.

CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) is a zwitterionic surfactant, which has been extensively used in various biological fields. In the present work, the concentration-dependent aggregation of the surfactant in deuterium oxide solution was investigated by NMR spectroscopy. We have found that two break points exist on the basis of the NMR parameters such as chemical shift difference (Δδ), self-diffusion coefficient (D), and relaxation rates (R1, R2). The first break point corresponds to the widely accepted normal critical micelle concentration (cmc). The second one is unexpected and ascribed to the second cmc, indicating that there is another type of micelle at higher concentrations. Further analysis using 1D selective NOESY with spin-diffusion quenching reveals that at the concentration above the second cmc CHAPS may form a two-layer spherical structure of micelles, with the aliphatic groups of CHAPS molecules in the inner layer interact with the steroid groups in the outer layer. The existence of two types of micelles has also been supported by our TEM experiment. The dependence of CHAPS micelle size on concentration explains why some proteins are soluble and stable only at concentrations above the second cmc. Therefore, our finding provides a basis for optimizing CHAPS concentration in functional and structural studies of membrane proteins.

In order to identify the neurochemical alterations in motor associated subcortical nuclei, and enhance our understanding of neurophysiology of progressive reduction in fine motor control with aging, the metabolic changes in striatum and cerebellar cortex in SD rats along with aging were investigated using a metabonomic approach based on high resolution “magic angle spinning” 1H-NMR spectroscopy and partial least squares-discriminant analysis. It was found that there were increased myo-inositol and lactate, and decreased taurine in these two brain regions of old rats. The above changes may be a marker for alterations of neuronal cells, which reduce fine motor control. Besides, some of the metabolites are gender-related and region-specific. Old female rats had decreased glutamate and increased creatine in striatum, while old male rats had increased choline in striatum, and increased GABA in cerebellar cortex, respectively. However, further analyses showed that most of the metabolites in male rats were not distinctively different with those of female ones except choline, which was in a relative lower level in striatum of male rats. All this results suggest that energy metabolism is an important indication of age-related change, which is not only in male, but also in female rats.Highlights► Old rats had increased myo-inositol and lactate in striatum and cerebellar cortex. ► Old female rats had decreased glutamate and increased creatine in striatum. ► Choline in striatum and GABA in cerebellar cortex increased in old male rats. ► Energy metabolism is an important indication of age-related change.