Isolation and characterisation of quercitrin as a potent anti-sickle cell anaemia agent from Alchornea cordifolia.

Alchornea cordifolia MÜll. Arg. (commonly known as Christmas Bush) has been used traditionally in Africa to treat sickle cell anaemia (a recessive disease, arising from the S haemoglobin [Hb] allele) but the active compounds are yet to be characterised. Herein we describe the use of sequential fractionation coupled with in vitro anti-sickling assays to purify the active component. Sickling was induced in HbSS genotype blood samples using sodium metabisulphite (Na2S2O5) or incubation in 100 % N2. Methanol extracts of A. cordifolia leaves and its sub-fractions showed >70 % suppression of HbSS erythrocyte sickling. Purified compound demonstrated 87.2 ± 2.39 % significant anti-sickling activity and 93.1 ± 2.69 % erythrocyte sickling-inhibition at 0.4 mg/mL. Nuclear magnetic resonance (NMR) spectra and high-resolution mass spectroscopy identified it as quercitrin (quercetin 3-rhamnoside). Purified quercitrin also inhibited the polymerisation of isolated HbS and stabilized sickle erythrocytes membranes. Metabolomic comparisons of blood samples using flow-infusion electrospray-high resolution mass spectrometry indicated that quercitrin could convert HbSS erythrocyte metabolomes to be similar to HbAA. Sickling was associated with changes in anti-oxidants, anaerobic bioenergy, and arachidonic acid metabolism, all of which were reversed by quercitrin. The findings described could inform efforts directed to the development of an anti-sickling drug or quality control assessments of A. cordifolia preparations.

Screening of the Chemical Composition and Identification of Hyaluronic Acid in Food Supplements by Fractionation and Fourier-Transform Infrared Spectroscopy

Hyaluronic acid, together with collagen, vitamins or plant extracts, is a part of many cosmetic and food preparations. For example, this polysaccharide is used in formulation of many food supplements due to its protective effects on human health. In this work, the screening of the chemical composition of three chosen dietary supplements (powder, tablets and capsules) containing hyaluronic acid was carried out using Fourier-transform infrared spectroscopy. Because of the low amount of analyte in all these samples, it was isolated or concentrated prior to the analysis using a suitable sequential fractionation protocol. Individual isolation procedures were established for each sample based on their declared composition. Firstly, the major components such as collagen or vitamins were removed to obtain polysaccharide fractions by the enzymatic treatment and/or washing out with the appropriate solvents. In some cases, the water insoluble part was removed from the rest dissolved in water. Then, hyaluronic acid was precipitated with copper(II) cations and thus separated from the other polysaccharides. Finally, the analyte was identified in the enriched fractions by the characteristic vibrational bands. The amount of hyaluronic acid in the purified fractions was determined in three ways: gravimetrically, spectrophotometrically, and using isotachophoresis. The combination of the appropriate preparative and analytical steps led to the successful evaluation of chemical composition, finding and quantification of hyaluronic acid in all the studied samples.

Zinc Plant Uptake as Result of Edaphic Factors Acting

The influence of soil characteristics on the lability and bioavailability of zinc at both background and phytotoxic concentrations in Albic Retisol soil (Loamic, Ochric) was studied using various methods. Ranges of insufficient, non-phytotoxic, and phytotoxic zinc concentrations in soil solutions were established in an experiment with an aqueous barley culture. It was experimentally revealed that for a wide range of non-toxic concentrations of Zn in the soil corresponding to the indicative type of plant response, there was constancy of the concentration ratio (CR) and concentration factor (CF) migration parameters. As a result, a new method for assessing the buffer capacity of soils with respect to Zn (PBCZn) is proposed. The transformation processes of the chemical forms and root uptake of native (natural) zinc contained in the Albic Retisol (Loamic, Ochric) through the aqueous culture of barley were studied using a cyclic lysimetric installation and radioactive 65Zn tracer. The distribution patterns of Zn(65Zn) between different forms (chemical fractions) in the soil were established using the sequential fractionation scheme of BCR. The coefficients of distribution and concentration factors of natural Zn and 65Zn, as well as accumulation and removal of the metal by plants were estimated. The values of the enrichment factor of natural (stable) Zn contained in sequentially extracted chemical fractions with the 65Zn radioisotope were determined and the amount of the pool of labile zinc compounds in the studied soil was calculated.

Influence of Soil Properties on P Pools and Its Effect on Forest Productivity in Mediterranean Calcareous Soils

Although soil phosphorus is essential for vegetation growth it is not always found in labile forms, hampering its absorption by plants, and is limited in forest ecosystems. This study explores soil P state and availability in calcareous soils, determining which processes affect its different pools and which soil parameters influence forest productivity of a Mediterranean pine species. We used a sequential fractionation method to determine P fractions in the soil according to their lability and their organic or inorganic nature. Those fractions were related to different soil and climatic parameters and to the site index in 32 Pinus halepensis plots of the National Spanish Forest Inventory. Soil texture, carbonates content, water retention capacity and organic matter substantially affected P fractions in the calcareous soils studied. Membrane extractable P was mainly influenced by organic matter-related parameters while the predominant P fraction in these soils, the primary P, was linked to the carbonates content. The biological mineralization processes played a key role in the soil P cycle. Total P, carbonates content, soil texture and water availability drove forest productivity in the studied plots.

Physical-chemical characterization of uranium containing sediments

The presented paper is intended to study the chemical behaviour of combined geogenicanthropogenic uranium content in specific stream sediments. The sampling points have been chosen with respect to the natural conditions in the locality of groundwater outflow from a former uranium mine adit in Považský Inovec mountain range, near Kálnica village. Besides the total uranium determination and physical-chemical characterization of the relevant water- and sediment samples we carried out modified Tessier type sequential fractionation extractions of natural- and artificially contaminated sediment samples after time dependent agitation in air and in the atmosphere of argon. The obtained results have been compared with those fulfilled with montmorillonite K-10. The total uranium concentrations of the relevant groundwater samples as well as of stream sediments have been determined by ICP-MS using HP 4500. The determinations of uranium in extracts have been accomplished according STN757614, utilizing arsenazo III as a selective complex forming reagent for spectrophotometric determination of uranyl-ions at 650 nm. The total uranium concentration of the groundwater outflow and in the sediment taken in its immediate vicinity has been 31.75±0.35 μg dm-3 and 38.0±2.7 μg g-1 respectively. Unlike montmorillonit K-10, in which the carbonate-bound fraction of uranium after 1 week aeration and agitation in argon atmosphere represents 22.8% and 18.6% respectively, uranium in investigated sediments has been present predominantly in carbonate-bound fraction-reaching under similar conditions 38.6% and 26.6%, respectively.

Terra Preta Properties in Northwestern Amazonia (Colombia)

Whereas many researchers still approach Terra Preta (TP) as a soil category, new evidence suggests that TP refers to a directional grading of soil property changes (i.e., color, pH, nutrients, etc.) within human-made soils, originating from human activities in pre-Columbian times. Currently, most TP research focuses on the Brazilian part of the Amazon basin, but only little information is available on TP soils in the Colombian Amazon. Here, we sampled four TP and surrounding soils in the Colombian Amazon region at different soil depths and analyzed them for (i) general soil properties such as color, pH and texture, (ii) soil organic carbon and black carbon (BC) contents, the latter using benzene polycarboxylic acids as molecular marker, (iii) phosphorus availability based on sequential fractionation, and (iv) microbial residue contents using amino sugars. Our data from Colombia’s middle Caquetá River and Leticia confirmed that SOC, BC, and total P were present in significantly higher concentrations in the TP areas than the surrounding soils, while pH values and microbial residue contents were unchanged. The enrichment of P forms comprised both easily extractable and stable P pools, which both dominated to a different degree, both in TP and adjacent soils. The different degree of SOC, BC and P enrichment suggests different amounts of waste disposal by the ancient populations at different TP sites, now warranting further research for reconstructing ancient population sizes from TP chemical analyses.