Mass spectrometrically detected directly coupled high performance liquid chromatography/nuclear magnetic resonance spectroscopy/mass spectrometry for the identification of xenobiotic metabolites in maize plants

Reconstructed ion chromatograms have been used to identify relevant high performance liquid chromatography (HPLC) peaks in a directly coupled high performance liquid chromatography/nuclear magnetic resonance spectroscopy/mass spectrometry (HPLC/NMR/MS) experiment. This has been applied to a study of the metabolism of a model compound, 5-nitropyridone (2-hydroxy-5-nitropyridine), in maize plants grown hydroponically. By monitoring the on-flow reconstructed ion chromatogram corresponding to the 5-nitropyridone fragment at m/z 143, and additional molecular ions corresponding to metabolites identified as products from similar compounds, relevant peaks were identified rapidly for subsequent stopped-flow 1H NMR spectroscopic analysis. The combination of coupled HPLC/NMR/MS enabled the direct identification of three metabolites, namely the N-glucoside, N-malonylglucoside, and O-malonylglucoside. This work demonstrates the power of HPLC/NMR/MS for the structural elucidation of xenobiotic metabolites in complex biological matrices (such as plant material) with minimal sample preparation. In particular, using mass spectrometry for the initial identification of relevant HPLC peaks allows the analysis of complex samples without the necessity for other spectroscopic markers, such as 19F NMR signal for fluorinated compounds or UV spectroscopy for molecules with strong UV chromophores.     

Investigation of water environments in a C18 bonded silica phase using 1H magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy

High resolution 1H magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra have been obtained on typical C18 bonded silicas used in chromatographic solid-phase extraction separations. It has been shown for the first time that water molecules distributed in distinct physico-chemical environments within the chromatographic system can be detected directly using a simple 1H MAS NMR measurement. The resonances assigned to water protons in differing physico-chemical environments have distinct chemical shifts, line widths, relaxation times (T1 and T2) and also exhibit temperature dependent coalescence behaviour. This novel MAS approach may lead to a better understanding of the environments of other analytes in mixtures during such separations.     

Magnetic resonance imaging in a model of atherosclerosis: use of a collar around the rabbit carotid artery

Transverse cardiac-cycle gated high resolution magnetic resonance images have been obtained from the neck of the New Zealand white rabbit both in normal animals and from those in which a collar had been earlier positioned around one carotid artery. The study included animals fed on normal and on high cholesterol diets with the surgical modification having been demonstrated previously to cause a rapid and reproducible lesion resembling early atherosclerosis. The aim of the work was to investigate the attainable spatial resolution and sensitivity at a field strength of 2 T using a large radiofrequency transmitter system and a surface coil receiver with which spin-echo images have been obtained. Visualization was enhanced using a three-dimensional interpolation technique. An image resolution of 200 microns was readily obtained but was shown to be insufficient for delineating pathological features within the artery wall such as intimal layer thickening. The results have been compared with histopathological findings which confirmed that any morphological changes were within the pixel resolution of the image. Extensions to the methodology are proposed which should be able to detect atherosclerotic changes with a resolution of 50 microns within a feasible imaging time. In addition, the MRI study of how the surgical intervention alters the artery shape and curvature was carried out and the MRI demonstrated that collar implantation in general does not occlude the artery and causes only a slight and gradual degree of curvature to the vessel.     

Heritability of Stroop and flanker performance in 12-year old children

Background  

There is great interest in appropriate phenotypes that serve as indicator of genetically transmitted frontal (dys)function, such as ADHD. Here we investigate the ability to deal with response conflict, and we ask to what extent performance variation on response interference tasks is caused by genetic variation. We tested a large sample of 12-year old monozygotic and dizygotic twins on two well-known and closely related response interference tasks; the color Stroop task and the Eriksen flanker task. Using structural equation modelling we assessed the heritability of several performance indices derived from those tasks.