Environmentally Sustainable Achiral and Chiral Chromatographic Analysis of Amino Acids in Food Supplements

Two LC methods were developed for the achiral and chiral reversed-phase (RP) analysis of an amino acid (AA) pool in a food supplement, in compliance with the main paradigms of Green Chromatography. A direct achiral ion-pairing RP-HPLC method was optimized under gradient conditions with a water-ethanol (EtOH) eluent containing heptafluorobutyric acid (0.1%, v/v), to quantify the eight essential AAs (Ile, Leu, Lys, Met, Phe, Thr, Trp, and Val) contained in the food supplement. Thus, the usually employed acetonitrile was profitably substituted with the less toxic and more benign EtOH. The method was validated for Leu and Phe. The chiral LC method performed with a teicoplanin chiral stationary phase was developed with a water-EtOH (60:40, v/v) eluent with 0.1%, v/v acetic acid. The enantioselective analysis was carried out without any prior derivatization step. Both developed methods performed highly for all eight AAs and revealed that: (i) the content of six out of eight AAs was consistent with the manufacturer declaration; (ii) only L-AAs were present. Furthermore, it was demonstrated that a two-dimensional achiral–chiral configuration is possible in practice, making it even more environmentally sustainable. A molecular modelling investigation revealed interesting insights into the enantiorecognition mechanism of Lys.     

Optimization of DNA extraction from dental remains

Efficient DNA extraction procedures is a critical step involved in the process of successful DNA analysis of such samples. Various protocols have been devised for the genomic DNA extraction from human tissues and forensic stains, such as dental tissue that is the skeletal part that better preserves DNA over time. However DNA recovery is low and protocols require labor‐intensive and time‐consuming step prior to isolating genetic material. Herein, we describe an extremely fast procedure of DNA extraction from teeth compared to classical method. Sixteen teeth of 100‐year‐old human remains were divided into two groups of 8 teeth and we compared DNA yield, in term of quantity and quality, starting from two different sample preparation steps. Specifically, teeth of group 1 were treated with a classic technique based on several steps of pulverization and decalcification, while teeth of group 2 were processed following a new procedure to withdraw dental pulp. In the next phase, the samples of both group underwent the same procedure of extraction, quantification and DNA profile analysis. Our findings provide an alternative protocol to obtain a higher amount of good quality DNA in a fast time procedure, helpful for forensic and anthropological studies.     

Enzyme‐Mediated Preparation of Chiral 1,3‐Dioxane Odorants

The enantiomerically enriched diastereoisomers of the chiral 1,3‐dioxane odorants Floropal^® ( 1 ) and Magnolan^® ( 2 ) were prepared by an enzyme‐mediated approach. Their olfactory properties were evaluated to investigate differences in the odor perception for the stereoisomers.     

Metal‐ion complexes in the angiogenetic effect

The complex formation between two polysaccharides, hyaluronic acid (Hyal) and its sulphated derivative (HyalS), and two metal ions, Cu²⁺ and Zn²⁺ was investigated in aqueous solution by thermodynamic and spectroscopic techniques. A stoichiometry for the complex species in solution was obtained. The bioactivity of the metal‐polysaccharide complexes was then evaluated in terms of their influence on endothelial cell migration and adhesion. The biological response of the complex species was found to be dependent of both the polysaccharide (Hyal or HyalS) and the metal ion (Cu²⁺ or Zn²⁺).     

Two-step elution of human serum proteins from different glass-modified bioactive surfaces: a comparative proteomic analysis of adsorption patterns

Plasma protein adsorption patterns on surfaces may give vital information to evaluate biocompatibility of biomaterials designed for direct blood-contacting applications or tissue integration. Adsorption of human serum proteins on four different types of biomaterials (glass, aminosilanized glass, hyaluronan and sulfated hyaluronan) was analyzed by two-dimensional electrophoresis. Desorption of proteins from the surfaces was first classically achieved by sodium dodecyl sulfate (SDS) elution. We introduced a second elution step (by use of isoelectric focusing (IEF) sample buffer consisting of urea, 3-[(3-cholamidopropyl)dimethylammonio]-1-propansulfonate, and dithioerythritol) which allows more stringent elution conditions and is a tool to evaluate the protein adsorption strength to biomaterials. Moreover, the two-step elution may discriminate between irreversible and reversible adsorption of plasma proteins for biomaterials, thus helping to elucidate the structure of protein multilayers which form a complex system at the surfaces. The IEF sample buffer proved not to alter the biomaterial structure and integrity. Hydrophobic bonds resulted to be the main strength driving protein adsorption onto our biomaterials. Apolipoproteins were the most important proteins interacting with the surfaces suggesting that high-density lipoprotein (HDL) particles could play a role in biocompatibility due to their beneficial effects on endothelial cells.     

Determination and assessment of the contents of essential and potentially toxic elements in Ayurvedic medicine formulations by inductively coupled plasma-optical emission spectrometry

Traditional Ayurvedic remedies are easily available nowadays not only in India, their country of origin, but also in Western countries. Some of these products contain high concentrations of potentially toxic elements as main or secondary ingredients, in addition to elements essential for human health; for these reasons, it is interesting to determine their elemental composition. In this study we assessed the concentrations of fifteen elements (Al, As, Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Pb, Si and Zn) in five products of the Parpati family, a group of Ayurvedic medicines containing high concentrations of mercury, manufactured in various places in India. Concentrations were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES) or (for Pb and Cd) by graphite furnace atomic absorption spectrometry (GF-AAS) after sample mineralization. We compared the calculated daily intake of each element with reference values, considering maximum tolerable intake levels or recommended nutrient amounts. The experimental results were treated with chemometric pattern recognition techniques. We found differences in the composition of products of the same denomination manufactured by different companies and strong correlations among groups of variables. As expected, the daily intake of mercury upon consumption of Parpati medicines largely exceeded the tolerable intake level of this element.     

The role of chemometrics in single and sequential extraction assays: a review. Part II. Cluster analysis, multiple linear regression, mixture resolution, experimental design and other techniques

Single and sequential extraction procedures are used for studying element mobility and availability in solid matrices, like soils, sediments, sludge, and airborne particulate matter. In the first part of this review we reported an overview on these procedures and described the applications of chemometric uni- and bivariate techniques and of multivariate pattern recognition techniques based on variable reduction to the experimental results obtained. The second part of the review deals with the use of chemometrics not only for the visualization and interpretation of data, but also for the investigation of the effects of experimental conditions on the response, the optimization of their values and the calculation of element fractionation. We will describe the principles of the multivariate chemometric techniques considered, the aims for which they were applied and the key findings obtained. The following topics will be critically addressed: pattern recognition by cluster analysis (CA), linear discriminant analysis (LDA) and other less common techniques; modelling by multiple linear regression (MLR); investigation of spatial distribution of variables by geostatistics; calculation of fractionation patterns by a mixture resolution method (Chemometric Identification of Substrates and Element Distributions, CISED); optimization and characterization of extraction procedures by experimental design; other multivariate techniques less commonly applied.     

N-Demethyl-sauroxine,a novel Lycodine Group alkaloid from Huperzia saururus

The present study describes the isolation and identification of N-demethyl-sauroxine, a novel Lycopodium alkaloid obtained from Huperzia saururus (Lam.) Trevis. (Lycopodiaceae). Its structure and relative stereochemistry were elucidated on the basis of its spectral data and chemical correlations. Additionally, acetylcholinesterase inhibitory activity was evaluated (IC₅₀ = 209.6 ± 1.1 μM). The structure of the already identified alkaloid sauroxine was also re-validated through two dimensional NMR data.