Anna Poliwoda

Supported liquid membrane extraction with single hollow fiber for the analysis of fluoroquinolones from environmental surface water samples.

In this work, the simple analytical method for the determination of four fluoroquinolone antibiotics: ciprofloxacin, enrofloxacin, norfloxacin and danofloxacin, in environmental surface water samples is described. Sample pretreatment step was performed by the application of a technique based on supported liquid membrane extraction with the configuration of single hollow fiber (HF-SLM). The HPLC system with diode array detection was used for final analysis of studied analytes. Various parameters affecting the extraction efficiency during HF-SLM enrichment, such as type of membrane diluent, pH of donor (sample) and acceptor phases, as well as an enrichment time and salt content of sample were studied. Using the presented hollow-fiber extraction high recovery (70-80%) was achieved. It gave enrichment factor above 100. The detection limits in surface water samples, for the four target antibiotics, were at range 0.01-0.02 microg/l, when 10 ml samples were processed. The obtained results demonstrate the applicability of presented method for the selective extraction of fluoroquinolones in environmental water samples at ultratrace level. Errors, expressed as relative standard deviation (RSD) were below 8%, for all tested concentration levels.

Surface molecularly imprinted silica for selective solid-phase extraction of biochanin A, daidzein and genistein from urine samples.

Selective molecularly imprinted silica polymer (SiO2MIP) for extraction of biochanin A, daidzein and genistein was synthesized using the surface molecular imprinting technique with the silica gel as a support. Biochanin A (BCA) was used as a template, 3-aminopropyltriethoxysilane (APTES) as a functional monomer, and tetraethoxysilicane (TEOS) as a cross-linker. Non-imprinted polymer with the sol-gel process (SiO2NIP) was also prepared for comparison. The synthesized polymers were characterized by Fourier transform infrared spectrometry (FTIR), scanning electron microscopy (SEM) and a standard Brunauer-Emett-Teller (BET) and Barret-Joyner-Halenda (BJH) analysis. The obtained results indicated the structural differences between imprinted and non-imprinted polymers. Finally, the SiO2MIP and SiO2NIP were adopted as the adsorbents of solid phase extraction for isolation and preconcentration of biochanin A and its structural analogues-daidzein and genistein from aqueous and urine samples. The performance analysis revealed that SiO2MIP displayed better affinity to the three investigated isoflavones compared with SiO2NIP. The recoveries of spiked samples for studied analytes ranged from 65.7% to 102.6% for molecularly imprinted silica polymer and 8.9-16.0% for non-imprinted sorbents.

Sample pretreatment techniques for oligopeptide analysis from natural sources

The analysis of oligopeptides in samples of food, tissues, and body fluids attracts considerable attention. The complexity of such samples requires efficient sample preparation (i.e., concentration and cleanup) procedures to remove interfering endogenous compounds and inorganic or organic salts. The methods of sample pretreatment that enable effective and selective isolation and/or preconcentration of oligopeptides from complex sample matrices have been reviewed. In each case, examples of application were presented and discussed, taking into account selectivity, enrichment, method automation, cleanup, and environmental aspects of the developed methods.

Multivariate optimization of the hollow fibre liquid phase microextraction of muscimol in human urine samples

A liquid phase microextraction based on hollow fibre followed by liquid chromatographic determination was developed for the extraction and quantitation of the hallucinogenic muscimol from urine samples. Method applicability on polar hallucinogens was also tested on two alkaloids, a psychedelic hallucinogen, tryptamine and a polar amino acid, tryptophan which exists in its charged state in the entire pH range. A multivariate design of experiments was used in which a half fractional factorial approach was applied to screen six factors (donor phase pH, acceptor phase HCl concentration, carrier composition, stirring rate, extraction time and salt content) for their extent of vitality in carrier mediated liquid microextractions. Four factors were deemed essential for the effective extraction of each analyte. The vital factors were further optimized for the extraction of single-spiked analyte solutions using a central composite design. When the simultaneous extraction of analytes was performed under universal factor conditions biased towards maximizing the enrichment of muscimol, a good composite desirability value of 0.687 was obtained. The method was finally applied on spiked urine samples with acceptable enrichments of 4.1, 19.7 and 24.1 obtained for muscimol, tryptophan and tryptamine respectively. Matrix-based calibration curves were used to address matrix effects. The r(2) values of the matrix-based linear regression prediction models ranged from 0.9933 to 0.9986. The linearity of the regression line of the matrix-based calibration curves for each analyte was directly linked to the analyte enrichment repeatability which ranged from an RSD value of 8.3-13.1%. Limits of detection for the developed method were 5.12, 3.10 and 0.21ngmL(-1) for muscimol, tryptophan and tryptamine respectively. The developed method has proven to offer a viable alternative for the quantitation of muscimol in human urine samples.

Do Differences in Chemical Composition of Stem and Cap of Amanita muscaria Fruiting Bodies Correlate with Topsoil Type

Fly agaric (Amanita muscaria) was investigated using a 1H NMR-based metabolomics approach. The caps and stems were studied separately, revealing different metabolic compositions. Additionally, multivariate data analyses of the fungal basidiomata and the type of soil were performed. Compared to the stems, A. muscaria caps exhibited higher concentrations of isoleucine, leucine, valine, alanine, aspartate, asparagine, threonine, lipids (mainly free fatty acids), choline, glycerophosphocholine (GPC), acetate, adenosine, uridine, 4-aminobutyrate, 6-hydroxynicotinate, quinolinate, UDP-carbohydrate and glycerol. Conversely, they exhibited lower concentrations of formate, fumarate, trehalose, α- and β-glucose. Six metabolites, malate, succinate, gluconate, N-acetylated compounds (NAC), tyrosine and phenylalanine, were detected in whole A. muscaria fruiting bodies but did not show significant differences in their levels between caps and stems (P value>0.05 and/or OPLS-DA loading correlation coefficient <0.4). This methodology allowed for the differentiation between the fruiting bodies of A. muscaria from mineral and mineral-organic topsoil. Moreover, the metabolomic approach and multivariate tools enabled to ascribe the basidiomata of fly agaric to the type of topsoil. Obtained results revealed that stems metabolome is more dependent on the topsoil type than caps. The correlation between metabolites and topsoil contents together with its properties exhibited mutual dependences.

Determination of muscimol and ibotenic acid in mushrooms of Amanitaceae by capillary electrophoresis

In this study, the CZE method for rapid quantitative and qualitative determination of ibotenic acid and muscimol in Amanita mushrooms naturally grown in Poland was developed. The investigations included the species of A. muscaria, A. pantherina, and A. citrina, collected in southern region of Poland. The studied hallucinogenic compounds were effectively extracted with a mixture of methanol and 1 mM sodium phosphate buffer at pH 3 (1:1 v/v) using ultrasound‐assisted procedure. The obtained extracts were separated and determined by CZE utilizing a 25 mM sodium phosphate running buffer adjusted to pH 3 with 5% content of acetonitrile v/v. The calibration curves for both analytes were linear in the range of 2.5–7000 μg/mL. The intraday and interday variations of quantitative data were 1.0 and 2.5% RSD, respectively. The recovery values of analyzed compounds were over 87%. The identities of ibotenic acid and muscimol were confirmed by UV spectra, migration time, and measurements after addition of external standard.

An Adventive Panaeolus antillarum in Poland (Basidiomycota, Agaricales) with Notes on Its Taxonomy, Geographical Distribution, and Ecology

Abstract Coprophilous fungus, Panaeolus antillarum rarely recorded in Europe, is reported here for the first time from the Augustów Plane, north-eastern Poland. This thermophilic species was found outdoors in August on horse dung mixed with straw. A chemical analysis did not confirm the presence of the psychoactive alkaloids in collected material. A complete description and illustration of the species based on Polish specimens are presented and notes on its taxonomy, ecology, world distribution and comparison with similar taxa — P. semiovatus var. semiovatus, P. semiovatus var. phalaenarum, and others are also provided.

Application of Molecular Imprinted Polymers for Selective Solid Phase Extraction of Bisphenol A

Abstract Selective molecularly imprinted polymers (MIPs) with bisphenol A as template were synthesized using the non-covalent imprinting approach. MIPs were prepared using thermally initiated polymerization with 1,1’-azobis(cyclohexanecarbonitryle) (ACHN) as initiator and ethylene glycol dimethacrylate (EDMA) as a cross-linking agent. The tested functional monomers included methacrylic acid, acrylamide, and 4-vinylpyridine. The selectivity of the BPA-MIP for the solid phase extraction of bisphenol A was tested in samples containing other related alkylphenols. The polymers prepared in acetonitrile using methacrylic acid or acrylamide as monomer showed the highest selectivity towards target analyte (the selectivity ratio 8:1, respectively for MIP and NIP). The proposed procedure has been proven to be an effective for selective extraction of bisphenol A in aqueous samples (recoveries over 85%) enabling detection and quantification limits of 25 and 70 μg/dm3, respectively based on 10 cm3 of sample volume, with relative standard deviations (RSD) lower than 6%. The obtained molecularly imprinted material showed interesting properties for selective extraction and preconcentration of studied analyte from large volumes of aqueous samples without any problems of cartridge clogging.

Chapter 5 - Bioactive Alkaloids of Hallucinogenic Mushrooms

The aim of our chapter is to review recent developments in a group of medicinally important natural products–alkaloids, with reference to the structure–activity studies in respect of certain diseases. Alkaloids covered by our review come from mushrooms called “hallucinogenic.” Hallucinogenic compounds have been chemically identified in mushrooms belonging to various genera, e.g., Agrocybe, Amanita, Conocybe, Galerina, Gymnopilus, Hypholoma, Inocybe, Panaeolus, Psilocybe, Pholiotina, Pluteus, and Weraroa [J.W. Allen, Ethnomycol. J. Sacred Mushroom Stud. 9 (2012) 130–175]. One of the largest classes of alkaloids is indole alkaloids. Indoles are probably the most widely distributed heterocyclic compounds in nature having medicinal importance [K.N. Kumar et al., Molecules 18 (2013) 6620–6662]. Two of simple indole alkaloids: psilocin (3-[2 (dimethylamino)ethyl]-4-indolol) and psilocybin ([3-(2-dimethylaminoethyl)-1H-indol-4-yl] dihydrogen phosphate) are present in most psychedelic mushrooms. Psilocin is a serotonin agonist – psilocybin/psilocin caused effects are thought to be mediated mainly by activation of 5-HT2A receptor. Ligands for the 5-HT2A receptor may be extremely useful tools for future cognitive neuroscience research [D.E. Nichols, Pharmacol. Ther. 101 (2004) 131–181]. They are also other analogs of psilocybin: baeocystin, norbaeocystin, bufotenin, and aeruginascin that are found in hallucinogenic mushrooms. Bufotenin occur also in some animal species (genus Bufo) and plants. Some of the mushroom belonging to genera Gymnopilus and Pholiota were shown to possess bisnoryangonin and hispidin, alkaloids with antimicrobial [K. Shinto et al., J. Home Econ. Jpn. 58 (9) (2007) 563–568] and antioxidant [Lee In-K et al., Mycobiology, 36 (1) (2008) 55–59] activity. Also psychoactive species of genus Amanita, contain the alkaloids (muscimol, ibotenic acid, and muscazone) that react with neurotransmitter receptors in the central nervous system. Isoxazoles, to which these alkaloids belong, have been found to inhibit voltage-gated sodium channels to control pain, enable the construction of tetracycline antibiotic derivatives, and as treatments for depression. The information within this review is intended to serve as a reference tool to better enable future research into important and fascinating area of pharmacognostic science as well as other parts of medical science.

Antioxidant Phenolic Compounds in Salvia officinalis L. and Salvia sclarea L.

Abstract The differences in the chemical composition of the phenolic compounds of Salvia officinalis versus Salvia sclarea growing in different habitats, were studied. First, the optimal solvent - methanol - for ultrasonic extraction of phenolic compounds from these plants was chosen experimentally. Total phenolic content and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays were determined spectrophotometrically. Total phenolic content was in the range of 63.9 to 134.4 mg GAE/g of plant depending on the studied species. The highest antiradical activity was displayed by the methanol fractions of S. sclarea varieties (83 and 67%). HPLC-DAD analysis of extracts was done in order to identify the presence of individual phenolic compounds. This was done by comparison of their retention times with those observed for 17 commercially available standard compounds. The results showed differences in the phenolics composition, with plants collected on Crimean peninsula accumulating more phenolic acids than Polish varieties. In turn, the Polish varieties of sage contain bigger amount of flavonoids in their tissues.