Malgorzata Szultka-Mlynska

Study of silver nanoparticles synthesized by acidophilic strain of Actinobacteria isolated from the of Picea sitchensis forest soil

In the present work the acidophilic actinobacteria strain was used as a novel reducing agent for the cheap, green and single‐step synthesis of nanostructure silver particles. Structural, morphological and optical properties of the synthesized nanoparticles have been characterized by spectroscopy, dynamic light scattering and electron microscopy approach. The antimicrobial activity of silver nanoparticles against clinical strains such as Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis and Salmonella infantis alone and in combination with antibiotics were studied.

Nanoporous Conducting Polymer-Based Coatings in Microextraction Techniques for Environmental and Biomedical Applications.

ABSTRACT Biologically active compounds constitute a wide group of chemicals, therefore it is a big challenge to create sorbents that are sensitive as well as selective. Development of nanoporous sorbents based on conducting polymers has expanded the boundaries of detection and quantification. Additionally, electrochemical synthesis used to deposit polymeric coatings directly on solid supports makes it possible to control physico-chemical properties of such sorbents. Besides the development of new polymeric nanoporous materials, the question of selectivity needs to be addressed. One possibility, successfully adapted to solid-phase microextraction, is molecular imprinting. Coatings created with this technology allow obtaining higher selectivity, with sensitivity at a constantly high level. The main aim of this review is to present comprehensively the concept of nanoporous sorbents based on conducting polymers, possible coating methods with their characteristics, and their various applications. This article focuses on applications in environmental and biomedical analyses.

Electrochemistry-mass spectrometry for in-vitro determination of selected chemotherapeutics and their electrochemical products in comparison to in-vivo approach.

Chemotherapeutics are among the most frequently prescribed medications in modern medicine. They are widely prescribed; however, problems with organisms developing resistance to these drugs means that their efficacy may be lost, so care should be taken to avoid unnecessary prescription. It is therefore of great interest to study the detailed metabolism of these biologically active compounds. This study aimed at developing an efficient analytical protocol for the determination of in-vitro electrochemical products of selected antibiotic drugs (amoxicillin, cefotaxime, fluconazole, linezolid, metronidazole and moxifloxacin). Combination of electrochemistry (EC) and mass spectrometry (MS) was applied for the in-vitro determination of the studied antibiotics and their electrochemical products. To identify the structure of the detected electrochemical products, MS/MS experiments were performed. This was one of the first applications of the EC system for generation of electrochemical products produced from antibiotic drugs. Adjustment of appropriate conditions and such parameters as the potential value, mobile phase (pH), working electrode and temperature had significant influence on electrochemical simulations and the creation of selected derivatives. Consequently, several working electrodes were evaluated for this purpose. In most of the studied cases, mainly two types of products were observed. One corresponded to an increase in mass by 14Da, which can be explained by a process consisting of oxidation (+16 m/z) and dehydrogenation (-2 m/z); The second in turn showed mass reduction by 14Da, which can be attributed to the loss of -CH2 as a result of N-demethylation. The performed experiments consisted of two stages: electrochemical oxidation of the analyzed samples (phase I of metabolic transformation), and addition of glutathione (GSH) for follow-up reactions (phase II conjunction). The electrochemical results were compared to in-vivo experiments by analyzing urine samples from patients after antibiotic drugs have been administered.. Overall, the comparison of electrochemistry to in-vivo experiments shows the high potential of EC-MS as a fast analytical tool in the prediction of electrochemical conversion that could be applied to therapeutic drug monitoring and pharmacokinetic studies as well.

The influence of different pH on the electrophoretic behaviour of Saccharomyces cerevisiae modified by calcium ions

The effect of a different pH on Saccharomyces cerevisiae cells modified with calcium ions was investigated by the capillary zone electrophoresis technique. For the identification of the wild strain of S. cerevisiae, the ribosomal nucleic acid sequencing and internal transcribed spacer sequencing as well as spectrometric approach were applied. The potentiometric titration and Fourier transform infrared spectroscopy have shown the occurrence of active functional groups such as carboxyl, amine/hydroxyl, phosphate/hydrogen phosphate groups on the surface of native yeast cells. Moreover, the spectroscopy study in a medium infrared range was carried out to identify the functional groups of yeast cells that participate in calcium ions binding interaction. Furthermore, the microscopic and spectrometric analysis shows that the pH value of the calcium ions solution has a significant effect on the intensity yeast cells clumping. Additionally, the impact of yeast cell clumping on the electrophoretic behaviours was examined. The modification of surface functional groups by calcium ions significantly affected the efficiency of electrophoretic separation. However, these changes did not affect the accuracy of S. cerevisiae identification by MALDI equipment with BioTyper platform. These results form the analytical solution for coupling of electrophoresis and MALDI-TOF MS technique.

Application of solid phase microextraction followed by liquid chromatography-mass spectrometry in the determination of antibiotic drugs and their metabolites in human whole blood and tissue samples

A sensitive, rapid and specific analytical method using high performance liquid chromatography coupled with mass spectrometry (HPLC-QqQ-MS) was developed to determine selected antibiotic drugs and their metabolites (amoxicillin, cefotaxime, ciprofloxacin, clindamycin and metronidazole; amoxycilloic acid, 4-hydroxyphenyl glycyl amoxicillin, desacetyl cefotaxime, 3-desacetyl cefotaxime lactone, ciprofloxacin N-oxide, N-demethylclindamycin, clindamycin sulfoxide, and hydroxy metronidazole) in human whole blood and vascularized tissue after single oral administration. The samples were prepared by solid phase microextraction with C18 fibers (SPMEC18) and determined on a GRACE analytical C18 column, Vision HT (50 × 2 mm, 1.5 μm) at the flow rate of 0.4 mL min-1 using water and acetonitrile (containing 0.1% formic acid) as the mobile phase. The proposed method was successfully applied in a pharmacokinetic study of the selected antibiotic drugs and their metabolites in real human samples. Additionally, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF-MS) was used for identification and qualification analysis of the target compounds.

Chromatographic behavior of selected antibiotic drugs supported by quantitative structure-retention relationships

As antibiotic drugs have a wide variety of applications, there is a growing demand for their selective determination, a task for which chromatographic methods seem to be appropriate. With this end in view, chromatographic measurements were performed with the use of six different HPLC columns (ACE 5C18, IAM.PC.DD.2, octadecyl, phenyl, alkylamide and cholesteryl-alkylamide) to determine the logkw of selected antibiotic drugs (amoxicillin, cefatoxime, ciprofloxacin, fluconazole, gentamicin, clindamycin, linezolid and metronidazole). The retention behavior of the analytes was investigated as a function of different binary hydro-organic mobile phases containing 10-45% (v/v) acetonitrile. The studied compounds were separated under isocratic conditions. The best results of separation of the tested biologically active compounds were obtained on the commercially available C18 column. Special attention was dedicated to the study of interactions among the stationary phase, mobile phase and the analytes. Another goal was to selecting the best column for separation of the tested biologically active compounds. Finally, QSRR models together with stationary phase characterization provided reliable information on the properties and characteristics of studied columns.

Structural characterization of electrochemically and in vivo generated potential metabolites of selected cardiovascular drugs by EC-UHPLC/ESI-MS using an experimental design approach

In the last few years, a number of studies were conducted which aimed at understanding the mechanisms of cardiovascular drug, metabolism, and there is still the need to determine the metabolites of cardiac drugs for the purpose of metabolism control. In this study, we employ a direct combination of electrochemical oxidation and mass spectrometric (EC-MS) identification for monitoring the oxidation pathway of ten cardiovascular drugs (metoprolol, propranolol, propafenone, mexiletine, oxprenolol, pirbuterol, pindolol, cicloprolol, acebutolol and atenolol). Oxidation was accomplished in an electrochemical thin-layer cell coupled on-line to electrospray ionization mass spectrometry (EC/ESI-MS). For further characterization of electrochemical products, the approach involving liquid chromatography linked to tandem mass spectrometry was used. Appropriate conditions for oxidation and identification processes with such parameters as the potential value, mobile phase (type and pH) and working electrode were optimized. Optimization was performed with the use of central composite design (CCD). Besides electrochemical oxidation of analytes (phase I of metabolic transformation), addition of glutathione (GSH) for follow-up reactions (phase II conjunction) was also investigated. The electrochemical results were compared to in-vivo experiments by analyzing plasma and urine samples from patients who had been administered selected cardiovascular drugs. These results show that electrochemistry coupled to mass spectrometry turned out to be an analytical tool suitable to procure a feasible analytical base for the envisioned in vivo experiments.

Electrochemical simulation of three novel cardiovascular drugs phase I metabolism and development of a new method for determination of them by liquid chromatography coupled with tandem mass spectrometry

In this study electrochemistry (EC) coupled with electrospray ionization mass spectrometry (ESI-MS) was used to study the metabolic fate of three novel cardiovascular drugs: rivaroxaban (RIV), aliskiren (ALS), and prasugrel (PRS). Mimicry of the oxidative phase I metabolism was achieved in a simple amperometric thin-layer cell equipped with a boron-doped diamond (MD) working electrode. Structures of the electrochemically-generated metabolites were elucidated from MS/MS experiments. Additionally, a sensitive, specific, and rapid ultra-high performance liquid chromatography-tandem mass spectrometer (UHPLC-MS/MS) method has been developed and validated for the selected drugs in human urine samples. Three different sample preparation methods were compared and finally, sample preparation was accomplished through an ultrasound-assisted emulsification microextraction process (USAEME). The drugs were detected using a triple quadrupole tandem mass spectrometer by multiple reaction monitoring via an electrospray ionization source with positive ionization mode (ESI(+)). The results obtained by EC-MS were compared with conventional in vivo studies by analyzing urine samples from patients. Data from in vivo experiments showed good agreement with the data from electrochemical oxidation. Thus, EC-MS is very well-suited for the simulation of the oxidative metabolism of rivaroxaban, aliskiren, and prasugrel as well. Moreover, electrochemical conversion of target compounds appears to be a new in vitro technology for the prediction of potential metabolites.

Antimicrobial Effectiveness of Bioactive Silver Nanoparticles Synthesized by Actinomycetes HGG16n Strain

BACKGROUND Biologically synthetized silver nanoparticles are promising antimicrobial agent. Flow cytometry, well diffusion methods, colony-forming units (CFU) and spectroscopic approach are commonly used in antimicrobial study. The aim of this study was investigation of effectiveness of Bio- AgNPs synthesized by Actinomycetes HGG16n using fluorescence flow cytometry method as an alternative to the standard ones (well and disc diffusion method). METHODS Flow cytometry technique was applied to monitor the antibacterial effect of biocolloidal silver and its combination with various commercial antibiotics against selected pathogens. The observed effectiveness was confirmed by fluorescence micrographs. RESULT Silver nanoparticles synthesized by Actinomycetes HGG 16n strain were as effective antimicrobial agent as the tested commercial antibiotics. Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Salmonella infantis and Bacillus subtilis strains are not able to create the resistant mechanisms under treatment of biocolloidal silver. Furthermore, the flow cytometry technique was more sensitive than disc and well diffusion and confirmed the effectiveness of BioAgNPs against all tested bacterial cells. Precipitation and limited diffusion of biocolloidal silver was observed by using well diffusion method. CONCLUSION Specificity and selection of antimicrobial approach are related with different nature of lowmolecular compounds (e.g. antibiotic) compared with biocolloids. An alternative method, flow cytometry was designed for antimicrobial study of biocolloidal silver nanoparticles and compared to the classical microbial techniques. Moreover, this work highlights the development of novel and inexpensive antimicrobial agent for most oral and skin infections caused by Gram (+) and Gram (-) bacteria.