Chemometrics-Based TLC and GC-MS for Small Molecule Analysis: A Practical Guide

Nowadays, thin-layer chromatography (TLC) and gas chromatography/mass spectrometry (GC-MS) instruments can produce more data than even before. At this point, the use of mathematical and statistical tools has provided the key to resolve the information overload. In this chapter, a practical guide is provided for the TLC and GC-MS analysis of short-chain fatty acids (SCFAs), amino acids, and monosaccharides. A methodology for extracting and transforming the chromatographic data to a suitable format for chemometrics is described. Furthermore, a procedure for chemometric analysis based on principal components analysis and clustering analysis is suggested

Carbohydrate Analysis by NIRS-Chemometrics

Near-infrared spectroscopy (NIRS) is a high-throughput, low-cost, solvent-free, and nondestructive analytical tool. Chemometrics is the science that employs statistical and mathematical methods to explain near-infrared spectra; it has been proven that when they are coupled, their effectiveness highly improved in-depth carbohydrate characterization. This chapter focuses on the fundamentals of near-infrared spectroscopy in the study of carbohydrates, as well as the application of partial least squares regression (PLSR) and principal component analysis (PCA), as the most useful chemometric techniques involved in carbohydrate analysis. The theoretical aspects and practical applications starting from simple to complex carbohydrates mixtures are covered. Indeed, the contributions from different fields extend the implementation of near-infrared spectroscopy from industrial quality control to scientific research

Cyclopalladated Compounds with Polyhalogenated Benzylphosphanes for the Mizoroki-Heck Reaction

Nine partially halogenated benzylphosphanes ArXCH2PR2 (ArX = 3,6-dichlorophenyl, 3,6- difluorophenyl and 3,4,5-trifluorophenyl; R = Ph, Cy, iPr) have been prepared and reacted with palladium acetate to obtain the cyclometallated dimers [Pd(μ-OAc)(κ2-C,P-ArXCH2PR2)]2. The acetate bridge has been exchanged by bromide using lithium bromide and the obtained dimers have been thoroughly characterised. The dimers with the non-halogenated phosphanes PhCH2PR2 (R = Ph, iPr) have also been prepared. Treatment with norbornadiene in the presence of silver tetrafluoroborate has furnished the cationic mononuclear complexes [Pd(κ2-C,P-ArXCH2PR2)(nbd)]BF4 as stable solids. These complexes and some of the bromide dimers have been used as catalytic precursors in the Mizoroki- Heck reaction between bromobenzene and butyl acrylate. The complexes efficiently catalyse this transformation and important differences of activity are found depending on the ligand. In general, fluorinated phosphanes give more active systems than chlorinated analogues

Accumulation and distribution of diterpenic acids in leaves of Montanoa tomentosa

Montanoa tomentosa has been used for at least last five centuries in traditional medicine in Mexico as a remedy for reproductive impairments. The accumulation of diterpenic acids in M. tomentosa leaves was determined. Using Scanning Electron Microscopy, the type and distribution of glandular trichomes (GTs) was observed on the abaxial and adaxial sides of leaves. GTs and non-glandular trichomes (NGTs) were observed on leaf surface, but the latter are confined to the leaves abaxial side. On the adaxial surface, only non-glandular trichomes were observed. Accumulation in GTs and leaf lamina of kaurenoic (KA) and grandiflorenic (GFA) acids was determined by Gas Chromatography coupled to an Electron Impact Mass Spectrometric Detector (GC/EI-MSD). GC-MSD analysis indicated that GTs accumulated KA and GFA, with KA accumulation being at a higher level than GFA in these structures. Attention on GFA and KA is due to their importance as plant growth regulator precursors with potential pharmacological applications.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Fecal Metabolomics Insights of Agavins Intake in Overweight Mice

Targeted and non-targeted metabolite profiling can identify biomarkers after a dietary treatment leading to a better understanding of interactions between diet and health. This study was conducted to establish enriched or depleted metabolites in the feces of overweight mice after a diet shift plus agavins or inulins supplementation, and their possible association with beneficial effects on host health. Thirty-eight male C57BL/6 mice were fed with a high-fat diet for 5 weeks followed by a diet shift to a standard diet supplemented with agavins (HF-ST + A) or inulins (HF-ST + I) for five more weeks. Feces were collected before and after prebiotic supplementation for metabolomics analyses. HF-ST + I group increased the fecal excretion of two methyl esters: linoleic and oleic acid, while HF-ST + A mice showed a substantial augment of 2-decenal, fructose, cyclohexanol, and the acids: 10-undecenoic, 3-phenyllactic, nicotinic, 5-hydroxyvaleric, and lactic. From the metabolites identified in HF-ST + A, only lactic acid has been reported previously and associated with beneficial effects on host health. However, the identification of new metabolites, coming from the microbial fermentation of agavins, opens opportunities to transform this information into practical solutions to tackle overweight and associated metabolic syndrome

Accumulation and distribution of diterpenic acids in leaves of Montanoa tomentosa

Montanoa tomentosa has been used for at least last five centuries in traditional medicine in Mexico as a remedy for reproductive impairments. The accumulation of diterpenic acids in M. tomentosa leaves was determined. Using Scanning Electron Microscopy, the type and distribution of glandular trichomes (GTs) was observed on the abaxial and adaxial sides of leaves. GTs and non-glandular trichomes (NGTs) were observed on leaf surface, but the latter are confined to the leaves abaxial side. On the adaxial surface, only non-glandular trichomes were observed. Accumulation in GTs and leaf lamina of kaurenoic (KA) and grandiflorenic (GFA) acids was determined by Gas Chromatography coupled to an Electron Impact Mass Spectrometric Detector (GC/EI-MSD). GC-MSD analysis indicated that GTs accumulated KA and GFA, with KA accumulation being at a higher level than GFA in these structures. Attention on GFA and KA is due to their importance as plant growth regulator precursors with potential pharmacological applications.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Molecular and Functional Characterization of Novel Fructosyltransferases and Invertases from Agave tequilana

Fructans are the main storage polysaccharides found in Agave species. The synthesis of these complex carbohydrates relies on the activities of specific fructosyltransferase enzymes closely related to the hydrolytic invertases. Analysis of Agave tequilana transcriptome data led to the identification of ESTs encoding putative fructosyltransferases and invertases. Based on sequence alignments and structure/function relationships, two different genes were predicted to encode 1-SST and 6G-FFT type fructosyltransferases, in addition, 4 genes encoding putative cell wall invertases and 4 genes encoding putative vacuolar invertases were also identified. Probable functions for each gene, were assigned based on conserved amino acid sequences and confirmed for 2 fructosyltransferases and one invertase by analyzing the enzymatic activity of recombinant Agave protein s expressed and purified from Pichia pastoris. The genome organization of the fructosyltransferase/invertase genes, for which the corresponding cDNA contained the complete open reading frame, was found to be well conserved since all genes were shown to carry a 9 bp mini-exon and all showed a similar structure of 8 exons/7 introns with the exception of a cell wall invertase gene which has 7 exons and 6 introns. Fructosyltransferase genes were strongly expressed in the storage organs of the plants, especially in vegetative stages of development and to lower levels in photosynthetic tissues, in contrast to the invertase genes where higher levels of expression were observed in leaf tissues and in mature plants

Synthesis, Characterization, and Biological Activity Studies of Copper(II) Mixed Compound with Histamine and Nalidixic Acid

A mixed copper complex with deprotonated nalidixic acid (nal) and histamine (hsm) was synthesized and characterized by FTIR, UV-Vis, elemental analysis, and conductivity. The crystal structure of [Cu(hsm)(nal)H2O]Cl·3H2O (chn) showed a pentacoordinated cooper(II) in a square pyramidal geometry surrounded by two N atoms from hsm, two O atoms from the quinolone, and one apical water oxygen. Alteration of bacterial DNA structure and/or associated functions in vivo by [Cu(hsm)(nal)H2O]Cl·3H2O was demonstrated by the induction of a recA-lacZ fusion integrated at the amyE locus of a recombinant Bacillus subtilis strain. Results from circular dichroism and denaturation of calf thymus DNA (CT-DNA) suggested that increased amounts of copper complex were able to stabilize the double helix of DNA in vitro mainly by formation of hydrogen bonds between chn and the sugars of DNA minor groove. In vivo and in vitro biological activities of the chn complex were compared with the chemical nuclease [Cu(phen)(nal)H2O]NO3 · 3H2O (cpn) where phen is phenanthroline

Synthesis and pharmacology of alkanediguanidinium compounds that block the neuronal nicotinic acetylcholine receptor

Taking as models the polyamine toxin fraction FTX from the funnel-web spider venom, and the guanidinium moiety of guanethidine, a series of azaalkane-1,omega-diguanidinium salts were obtained. Some of them blocked ion fluxes through the neuronal nicotinic receptors for acetylcholine (nAChR). The blockade was exerted at submicromolar concentrations, suggesting a highly selective interaction with the nAChR. In fact, the active compounds on the nAChR ion channel did not recognize the voltage-dependent Na+ or Ca2+ channels of bovine adrenal chromaffin cells. Therefore, these compounds may be useful tools to clarify the functions of nAChR receptors in the central and peripheral nervous systems.Fundación Ramón Arece