Genotoxicity of selected pharmaceuticals, their binary mixtures, and varying environmental conditions – study with human adenocarcinoma cancer HT29 cell line

Pharmaceutical residues are present in the environment in mixtures and their adverse effects may also result from interactions that occur between compounds. Studies presented in this work focus on genotoxicity of pharmaceuticals from different therapeutic groups in mixtures and in individual solutions impacted with different environmental conditions assessed using comet assay (alkaline approach). Binary mixtures of pharmaceuticals (in different concentration ratios) and in individual solutions impacted with pH change (range from 5.5 to 8.5) or addition of inorganic ions, were incubated with HT29 cells and after 24 h time period cells were tested for the presence of DNA damage. To estimate whether mixtures act more (synergistic) or less (antagonistic) efficiently Concentrations Addition (CA) and Independent Action (IA) approaches were applied followed by a calculation of the Model Deviation Ratio (MDR) to determine deviation from the predicted values. Addition of inorganic ions mainly reduced their genotoxicity. Diclofenac s. was the most susceptible to potassium, fluoride, and bromide ions. Change of the pH of pharmaceutical solutions had significant impact on genotoxicity of diclofenac s. and fluoxetine h. Among mixtures, more commonly observed interactions were synergistic ones, exactly twenty-five cases (ten pairs containing chloramphenicol or oxytetracycline h.) and ten cases of antagonism (four for pairs containing chloramphenicol or fluoxetine h.). The results obtained indicate that interactions between tested compounds occur frequently and can lead to DNA damage. This topic especially concerning in vitro tests using cells is still rare, however, it should not be neglected.

Sample Preparation in Foodomics: Miniaturized Solid-Phase Extraction

Analytical chemists face a challenge to bring comprehensive information on a given food and biological sample by using the best available analytical techniques and meet the requirements of sustainable development and green chemistry at the same time. A key objective of this chapter is to review selected literature data on the utilization of solid-phase extraction techniques with special attention to their miniaturized modes in order to assess the safety, quality and nutritional value of foods. The techniques will be discussed in the context of different samples and analytical information which is about to be obtained.

Detection, identification and determination of chiral pharmaceutical residues in wastewater: Problems and challenges

Chiral pharmaceuticals (CPs) are widely used in different areas of human life, thus they are frequentlydetected in different ecosystems. However, before CPs reach the environment, wastewater is subjected todifferent treatment processes in order to remove them. Nevertheless, such processes may affect thechirality of CPs, thus it is very important to monitor CP levels during the wastewater treatment.This review addresses the present state of knowledge concerning the input, occurrence, fate and ef-fects of CPs in the environment. It focuses primarily on wastewater analysis, problems and challengesconnected with trace levels of CP enantiomers and highly complex matrices of samples. Analytical ap-proaches used in detection, identification and determination of enantiomers are presented. The appli-cation of the results of wastewater analysis to obtain information on the population's health andbehaviour has been included and discussed. Moreover, the prospects of the future trends in greenenantiomeric analysis are described.

Determination of bromhexine and its metabolites in equine serum samples by liquid chromatography – Tandem mass spectrometry: Applicability to the elimination study after single oral dose

Bromhexine (BH), expectorant used in the treatment of respiratory disorders associated with viscid or excessive mucus, is not permitted for use in the competing horse by many authorities in horseracing and Olympic disciplines. Metabolic studies are of the great importance in anti-doping field because they allow for updating the selection of the most appropriate markers for prohibited substances, such as metabolites present at higher concentration levels and/or lasted for a longer period of time in biological samples than a parent drug. This study describes LC-MS/MS-based method for simultaneous determination of BH and its metabolites, including 4-(2-amino-3,5-dibromobenzylamino)cyclohexanol (4-HDMB), 3-(2-amino-3,5-dibromobenzylamino)cyclohexanol (3-HDMB), in equine serum samples. The 2-(2-amino-3,5-dibromobenzylamino)cyclohexanol (2-HDMB) was monitored as well. The assay was validated in terms of linearity (R2 greater than 0.9951), intra- and inter-assay accuracy (91.6 – 109.1%) and precision (CV < 9.6%) as well as recovery (94.8 – 105.65%). The LODs were 0.0052, 0.0053, 0.0056 and 0.0043 ng/mL for BH, 2-HDMB, 3-HDMB and 4-HDMB, respectively. The developed method was applied to determine the time curses of BH and its metabolites concentrations in equine serum collected for 95.25 h following a single oral administration of BH to two healthy mares (in dose of 0.8 mg/kg). The parent drug was found at higher concentration levels than 3-HDMB (major metabolite) and 4-HDMB (minor metabolite), however, both BH metabolites lasted for a longer period of time in equine serum than the parent drug. Thus, both metabolites of BH can be considered as BH abuse markers.

Development and validation of a GC–MS/MS method for the determination of 11 amphetamines and 34 synthetic cathinones in whole blood

M. Woźniak , L. Banaszkiewicz , M. Wiergowski, E. Tomczak, M. Kata, B. Szpiech, J. Namieśnik , M. Biziuk Forensic Toxicology – 2020
Purpose Psychoactive compounds that contain a phenylethylamine structure (such as amphetamine-type stimulants and synthetic cathinones) are one of the major classes of stimulants on the recreational drug market. Approximately 670 new psychoactive substances (NPS) are monitored only in Europe; however, new psychoactive compounds are being developed for illicit trade each year. In this context, the development of new analytical procedures for the determination of such compounds in biological specimens for forensic toxicology is of great importance. Methods Gas chromatography–tandem mass spectrometry (GC–MS/MS) technique was applied for analysis of amphetamines and synthetic cathinones. The volumes of 200 µL of each whole blood sample and 1 mL of liquid-liquid extraction solvent were used for extraction, followed by pentafuoropropionyl derivatization. Results A high-throughput, robust, rapid, and sensitive procedure involving a simple liquid-liquid extraction for the simultaneous determination of 45 amphetamine-type stimulants and synthetic cathinones in whole blood was developed. The assay was validated based on its recovery (83.2–106%), interday accuracy (89.0–108%), and interday precision (≤8.1%). In view of the low limits of detection (ranged between 0.02 and 0.72 ng/mL) and limits of quantifcation (1 and 2.5 ng/mL), the developed method can serve as a less expensive and more ecologically friendly alternative to the liquid chromatography–tandem mass spectrometric methods. Conclusions To the best of our knowledge, this is the frst work presenting a GC–MS/MS method for the determination of NPS in blood samples. The presented procedure was applied to authentic samples from forensic cases, demonstrating its utility in the quantifcation of a wide number of psychoactive substances in routine toxicological analyses. The developed procedure can also be easily expanded to additional compounds.

Evaluation of the Greenness of Analytical Procedures

This contributions introduces the need to develop the methods to evaluate analytical procedures in the light of green analytical chemistry. Green chemistry metrics are not applicable in analytical chemistry because they refer to the mass of product and no product with mass is generated during analytical determination. Analytical greenness evaluations are based on scoring - such as NEMI or Eco-scale or comparative analysis as it is in case of multicriteria decision analysis. Important aspect is also the assessment of solvents and reagents that are applied in analytical procedures for their greening. The different environmental, safety and health parameters are included in the assessments of solvents and reagents. The contribution presents the mechanisms of evaluation and numerous examples. Some results of reagents and solvents assessments are also presented to help analysts in selection greener alternatives.

Green Chromatography: State-of-the-art, Opportunities and Future Perspectives

It is well known that chromatographic procedures could have a significant impact on the environment if laboratory practice is not in line with the principles of green analytical chemistry (GAC). However, chromatographic techniques have the potential to be greener in all steps of the analysis. The approaches used to make chromatographic separations greener differ depending on the type of chromatographic method. This chapter considers the ways in which chromatography can become greener. Liquid and gas chromatography are compared from the GAC point of view. In addition, the importance of miniaturization in sample preparation and chromatographic separations is pointed out. On-line process analysers and portable chromatographs are also discussed.

Production of certified reference materials - homogeneity and stability study based on the determination of total mercury and methylmercury

Reference materials (RMs) play an important role in all elements of the quality assurance system of measurements. In this work, ``package'' 4 new CRMs (bottom sediment, herring tissue, cod tissue, cormorant tissue) were prepared and characterised to carry out the quality control in monitoring analysis of mercury and methylmercury in environmental samples. Materials - candidates were collected in Poland and south part of the Czech Republic. All materials were freeze-dried, milled, sieved, homogenised, sterilized and distributed in amber bottles. For the homogeneity and stability study of the samples of materials several statistical tests were applied. Materials - candidates for CRMs with certified mercury and methylmercury content meet the heterogeneity requirement and can be considered homogenous both between bottle and within bottle. Each material meets also the requirements for the stability condition of reference material with certified mercury content.

Pulsowo-różnicowa woltamperometria jako narzędzie pozwalające na wyznaczenie wartości mocy przeciwutleniającej

Standardowy potencjał redukcji (E0) jako parametr określający zdolność przeciwutleniaczy, jak np. flawan-3-oli do przyjmowania elektronów wykazywał wysoką korelację z ich aktywnością biologiczną. Jednak, w przypadku oceny całkowitej aktywności przeciwutleniającej mieszanin tych związków, parametr ten może okazać się być niewystarczającym. Bowiem do określenia tzw. mocy przeciwutleniającej ang. antioxidant power (AOP) powinny być brane pod uwagę trzy elementy takie jak: wielkość przenoszonego ładunku, czas jego przeniesienia oraz energia przenoszonego ładunku odzwierciedlająca potencjał piku utlenienia. Sumarycznie elementy te – jako wynik oznaczeń woltamperometrycznych, mogą zostać wyrażone w jednostce mocy prądu elektrycznego (wat) jako parametr mocy przeciwutleniającej. Spośród znanych technik woltamperometrycznych – pulsowo-różnicowa woltamperometria (DPV) charakteryzująca się wysoką czułością jest najbardziej odpowiednia do pomiaru całkowitej aktywności przeciwutleniającej próbki. Wartości AOP dla mieszanin flawan-3-oli, odzwierciedlające matrycę żywieniową – kakao, zostały otrzymane dwiema technikami: elektrochemiczną (DPV) i spektrofotometryczną z wykorzystaniem rodnika DPPH. Pomiary elektrochemiczne przeprowadzono w układzie trójelektrodowym, gdzie elektrodę pracującą stanowiła elektroda szklista węglowa. Otrzymane piki utlenienia – zależność zadanego potencjału od zmierzonego prądu, a także zaproponowany szereg obliczeń pozwoliły na wyznaczenie wartości AOP badanych próbek. Co z kolei pozwoliło na określenie efektu synergistycznego, antagonistycznego czy addytywnego poszczególnych składników matrycy żywieniowej.

Recent Applications of 1D GC-MS and 2D GC-MS in Foodomics Studies

Gas chromatography coupled with mass spectrometry and food chemistry have been tightly linked since the 1960s. This setup is also an integral part of the omics studies when it comes to low mass metabolites. Therefore, the marriage of GC-MS and foodomics is obvious. Two-dimensional gas chromatography coupled with mass spectrometry fits perfectly within the latest trends in foodomics, since it is tailored for holistic food analysis. In this article, the authors will focus on the current trends in 1D and 2D GC-MS-based foodomics.