Identification of New Hepatic Metabolites of Miconazole by Biological and Electrochemical Methods Using Ultra-High-Performance Liquid Chromatography Combined with High-Resolution Mass Spectrometry.

The main objective of this study was to investigate the metabolism of miconazole, an azole antifungal drug. Miconazole was subjected to incubation with human liver microsomes (HLM) to mimic phase I metabolism reactions for the first time. Employing a combination of an HLM assay and UHPLC-HRMS analysis enabled the identification of seven metabolites of miconazole, undescribed so far. Throughout the incubation with HLM, miconazole underwent biotransformation reactions including hydroxylation of the benzene ring and oxidation of the imidazole moiety, along with its subsequent degradation. Additionally, based on the obtained results, screen-printed electrodes (SPEs) were optimized to simulate the same biotransformation reactions, by the use of a simple, fast, and cheap electrochemical method. The potential toxicity of the identified metabolites was assessed using various in silico models.

Antifungal drugs in the aquatic environment: A review on sources, occurrence, toxicity, health effects, removal strategies and future challenges.

Fungal infections pose a significant global health burden, resulting in millions of severe cases and deaths annually. The escalating demand for effective antifungal treatments has led to a rise in the wholesale distribution of antifungal drugs, which consequently has led to their release into the environment, posing a threat to ecosystems and human health. This article aims to provide a comprehensive review of the presence and distribution of antifungal drugs in the environment, evaluate their potential ecological and health risks, and assess current methods for their removal. Reviewed studies from 2010 to 2023 period have revealed the widespread occurrence of 19 various antifungals in natural waters and other matrices at alarmingly high concentrations. Due to the inefficiency of conventional water treatment in removing these compounds, advanced oxidation processes, membrane filtration, and adsorption techniques have been developed as promising decontamination methods.In conclusion, this review emphasizes the urgent need for a comprehensive understanding of the presence, fate, and removal of antifungal drugs in the environment. By addressing the current knowledge gaps and exploring future prospects, this study contributes to the development of strategies for mitigating the environmental impact of antifungal drugs and protecting ecosystems and human health.

Electrochemical Simulation of Phase I Hepatic Metabolism of Voriconazole Using a Screen-Printed Iron(II) Phthalocyanine Electrode.

Understanding the metabolism of pharmaceutical compounds is a fundamental prerequisite for ensuring their safety and efficacy in clinical use. However, conventional methods for monitoring drug metabolism often come with the drawbacks of being time-consuming and costly. In an ongoing quest for innovative approaches, the application of electrochemistry in metabolism studies has gained prominence as a promising approach for the synthesis and analysis of drug transformation products. In this study, we investigated the hepatic metabolism of voriconazole, an antifungal medication, by utilizing human liver microsomes (HLM) assay coupled with LC-MS. Based on the obtained results, the electrochemical parameters were optimized to simulate the biotransformation reactions. Among the various electrodes tested, the chemometric analysis revealed that the iron(II) phthalocyanine electrode was the most effective in catalyzing the formation of all hepatic voriconazole metabolites. These findings exemplify the potential of phthalocyanine electrodes as an efficient and cost-effective tool for simulating the intricate metabolic processes involved in drug biotransformation, offering new possibilities in the field of pharmaceutical research. Additionally, in silico analysis showed that two detected metabolites may exhibit significantly higher acute toxicity and mutagenic potential than the parent compound.

Stability of aspartame in the soft drinks: Identification of the novel phototransformation products and their toxicity evaluation.

Photolytic transformation of aspartame - a widely used artificial sweetener - under the simulated sunlight was studied for the first time. The experiments were conducted in pH range of 2.5 - 7.0 and in eight soft drinks available in the market. The highest degradation rate in the tested buffered solutions was observed under the neutral pH conditions. Irradiation of the soft drinks resulted in significantly (up to tenfold) faster degradation of aspartame, regardless of its initial concentration in the beverage. Such considerable acceleration of decomposition, not reported for aspartame so far, was ascribed to influence of the co-occurring ingredients, which can act as the photosensitizers. These findings indicate that some formulations may be particularly unfavorable in the context of aspartame photostability. Qualitative analysis of the studied processes revealed formation of six phototransformation products including three previously not described. In silico estimation of toxicity showed that some of the identified photoproducts, including the novel phenolic derivatives, may be more harmful than the parent compound. Taking into account relatively extensive formation of those products in the soft drinks, such finding may be particularly important from the food safety point of view.

Multivariate Chemometric Comparison of Forced Degradation and Electrochemical Oxidation LC-MS Profiles of Maraviroc.

In this study, nine forced degradation products of maraviroc were found using chemometric analysis. This antiretroviral drug was subjected to photolytic, oxidative, as well as neutral, basic and acidic hydrolysis stress conditions. Additionally, its electrochemical transformation on platinum, gold and glassy carbon screen-printed electrodes was examined. This study showed that maraviroc is especially susceptible to UVA, H2O2 and electrochemical degradation, while being resistant to neutral and acidic hydrolysis. A cluster analysis showed that the electrochemical transformation, with particular reference to the platinum electrode, is able to partially simulate the forced degradation processes, especially in the context of redox reactions. These findings indicate that the electrochemical methods can be considered as quick and relatively low-cost supplements to the commonly applied forced degradation procedures.

Efficient removal of anti-HIV drug - maraviroc from natural water by peroxymonosulfate and TiO2 photocatalytic oxidation: Kinetic studies and identification of transformation products.

In this study photochemical transformation of the antiretroviral pharmaceutical maraviroc under the simulated UV-Vis radiation was presented. The drug was shown to be extremely photo-resistant, with a half-life over 250 h, which is particularly significant, considering its presence in the aquatic environments. Addition of the natural river water matrix substantially increased the degradation rate, albeit the process led to formation of numerous phototransformation products. Due to high photostability and presumable environmental persistence of maraviroc, a photocatalytic method of its elimination was proposed. Although titanium dioxide alone presented acceptable results, its combination with peroxymonosulfate enormously accelerated the degradation process, increasing it over 67 000 times in comparison with the direct photolysis. Substitution of ultrapure water with river water resulted in inhibition of the PMS-driven processes, however the decomposition efficiency was still very high. Noteworthy, majority of the identified photoproducts were still present after termination of irradiation in all the experiments, which may indicate necessity of ecotoxicological assessment of those compounds.

Identification of the New Metabolite of Nebivolol Using Liquid Chromatography Coupled with High-Resolution Mass Spectrometry and Chemometrics.

In this study, the phase I hepatic metabolism pathway of a cardiovascular drug nebivolol was proposed on the basis of a human liver microsomes assay with the use of LC-HR-MS coupled with the chemometric method. Six biotransformation products were found with the assistance of chemometric analysis. Five of them were identified as the previously reported products of alicyclic hydroxylation and dihydroxylation, aromatic hydroxylation, as well as alicyclic oxidation of the parent compound. Moreover, one metabolite, not reported so far, was found to be a product of N-dealkylation of nebivolol-2-amino-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl)ethan-1-ol. The novel metabolite was submitted to an in silico toxicity analysis to assess its biological properties. The applied computational methods indicated a significantly elevated risk of its mutagenic activity, compared to the parent molecule. Several metabolites of the nebivolol described in the literature were not detected in this study, indicating their non-hepatic origin.

Comparative analysis of in vivo and in silico toxicity evaluation of the organoiodine compounds towards D.magna using multivariate chemometric approach: A study on the example of amiodarone phototransformation products.

In the present study the photochemical fate of organoiodine compound - amiodarone was performed. The drug turned out to be highly susceptible to UV-Vis irradiation, especially in the presence of humic substances and organic matrix. Qualitative LC-MS analysis revealed formation of twelve - mainly previously unreported - transformation products (TPs). Four major TPs were submitted to the toxicity analysis with the use of D. magna. All of the tested TPs presented higher toxic potential than the parent compound. The phenolic TPs were approximately 100 times more toxic than amiodarone. Toxic properties of the major TPs resulted in steadily increasing toxic potential of the photo-generated mixture over the time of irradiation. Moreover, the experimental toxicity data, concerning the TPs, were compared with results estimated by 6 in silico models with the use of a multivariate chemometric analysis. The results showed that the applied computational methods were able neither to correctly predict toxic properties of the studied compounds, nor the trends in change of their toxic parameters. Additional validation of in silico models ability to predict toxicity of iodinated organic compounds showed that the studied computational methods do not present sufficient prediction ability. Therefore their estimations concerning organoiodines should be verified using experimental tests.

Exploring the Rare Etiology of Severe Anemia in an Immunocompromised Patient.

Pure red cell aplasia (PRCA) is a rare cause of profound anemia, marked by very low reticulocyte count and near to complete absence of erythroid precursor cells in the bone marrow. PRCA can be congenital such as in the case of children with Diamond- Blackfan anemia or acquired, which is often triggered by exposure to certain viruses or drugs. Management depends on the underlying etiology of PRCA. Here, we present the case of a young male with underlying acquired immunodeficiency syndrome, who presented with a hemoglobin of 2.6 g/dL, initially thought to be secondary to gastrointestinal blood loss, but was later discovered to have parvovirus-induced PRCA.

Characterization of Phase I Hepatic Metabolites of Anti-Premature Ejaculation Drug Dapoxetine by UHPLC-ESI-Q-TOF.

Determination of the metabolism pathway of xenobiotics undergoing the hepatic pass is a crucial aspect in drug development since the presence of toxic biotransformation products may result in significant side effects during the therapy. In this study, the complete hepatic metabolism pathway of dapoxetine established according to the human liver microsome assay with the use of a high-resolution LC-MS system was described. Eleven biotransformation products of dapoxetine, including eight metabolites not reported in the literature so far, were detected and identified. N-dealkylation, hydroxylation, N-oxidation and dearylation were found to be the main metabolic reactions for the investigated xenobiotic. In silico analysis of toxicity revealed that the reaction of didesmethylation may contribute to the increased carcinogenic potential of dapoxetine metabolites. On the other hand, N-oxidation and aromatic hydroxylation biotransformation reactions possibly lead to the formation of mutagenic compounds.

Photochemical transformation of fentanyl under the simulated solar radiation - Enhancement of the process by heterogeneous photocatalysis and in silico analysis of toxicity.

In this study the photochemical transformation of fentanyl-a very potent opioid drug-under simulated solar radiation was investigated for the first time. This pharmaceutical is frequently detected in various environment samples at concentrations that should be regarded as potentially harmful. Nevertheless, to date, no drug phototransformation products (TPs) have been reported. Considering fentanyl's exceptionally high toxicity, knowledge of the properties of these potential TPs is essential in order to properly assess its pollution impact. In this study, all photolytic experiments were performed using a xenon lamp (D65 filter) and RP-UHPLC coupled with the ESI-high-resolution tandem mass spectrometry. The phototransformation of fentanyl in natural river water and the application of heterogeneous photocatalysis as a possible way of decontaminating water were also investigated. Fentanyl turned out to be photostable, but twenty-six previously unreported TPs (formed mainly as a consequence of hydroxylation and oxidation) were found and characterized. The applied catalysts-TiO2 and ZnO-showed very high effectiveness, and the presence of the natural water matrix further increased the photodecomposition rate (up to 600 times) relative to direct photolysis. Importantly, the almost complete degradation of the parent compound as well as its TPs after 16 min of irradiation indicated that heterogeneous photocatalysis can be considered an efficient way of treatment of fentanyl-contaminated water. The computational analysis of toxicity showed that fentanyl may be more harmful to rodents and aquatic species than its TPs. However, some of these products are probably more mutagenic and developmentally toxic. Additionally, one product in particular may be a strong estrogenic compound, proving the importance of assessing TPs' toxic properties. The evaluation of bioaccumulation, bioconcentration and biodegradability revealed that fentanyl possesses unfavorable properties compared to TPs.

Influence of the UV-Vis irradiation on the acute toxicity to zebrafish and mutagenicity of the selected psychotropic drugs.

The influence of the UV-Vis radiation on the toxicity of agomelatine, loxapine, clozapine and tiapride was studied. The phototransformation procedure was conducted with the use of simulated solar radiation. In the case of each compound irradiation time necessary to decompose half of the initial concentration was chosen. The embryotoxicity and acute toxicity were evaluated using zebrafish (Danio rerio) embryos and larvae. The mutagenicity assay was done with the use of a micro-plate Ames test. Generally, the embryotoxicity decreased after the irradiation procedure. The obtained results showed that tiapride is the least toxic compound to zebrafish, however, its toxicity toward larvae increases after the irradiation. Similarly, the mutagenic potential of the mixture of tiapride photoproducts is higher than in the case of parent compound. The phototransformation of loxapine resulted in the change of the acute toxicity profile and increased the rate of reverse mutations in the Ames test. Oppositely, the irradiation of agomelatine caused the decrease of mutagenic potential and acute toxicity was also lower in the postirradiated mixture. The phototransformation of clozapine did not alter the mutagenicity and decreased the acute toxicity to the zebrafish larvae. In silico calculations showed a satisfactory prediction ability in some instances, especially in the case of mutagenic potential of the tiapride phototransformation products.

Identification and characterization of citalopram new metabolites with the use of UHPLC-Q-TOF technique: In silico toxicity assessment of the identified transformation products.

In this study the metabolite profiling of citalopram with the use of human liver microsomes as well as the complementary photocatalytic method were established. This strategy allowed the detection of five metabolites of citalopram including 3-hydroxycitalopram and 3-oxocitalopram which were found as a new and not previously described metabolites of this drug The photocatalytic simulation of metabolism was carried out using tungsten (VI) oxide nanopowders with the different particle sizes, which allowed to examine the effect of this photocatalyst parameter on the mapping of metabolic processes. The accurate characterization of all observed structures was possible due to the use of ultra-high-pressure liquid chromatography and high-resolution mass spectrometry combined system as a highly useful technique in drug metabolism studies. In order to perform the toxicity prediction of citalopram and its metabolites, the acute toxicity to rodents, as well as genotoxicity, carcinogenicity, developmental toxicity and receptor-mediated toxicity was calculated basing on the in silico tools.

Simulation of phase I metabolism reactions of selected calcium channel blockers by human liver microsomes and photochemical methods with the use of Q-TOF LC/MS.

The in vitro phase I metabolism of perhexiline and flunarizine, two calcium channel blockers was investigated during this study with the use of human liver microsomes (HLM) method compared with TiO2, WO3 and ZnO catalyzed photochemical reaction. In order to determine the structures of metabolites an quadrupole time-of-flight mass spectrometry combined with liquid chromatography (Q-TOF LC/MS) system was used. The obtained high resolution mass spectra enabled to identify thirteen products of metabolism of selected drugs including three not yet described metabolites of perhexiline and two new metabolites of flunarizine. The vast majority of metabolites were confirmed also with the participation of photocatalytic approach of the drug metabolism simulation. The comparison of all metabolic profiles made with the use of computational methods drew attention particularly to TiO2 and WO3 catalyzed photochemical reaction as similar to HLM incubation. Additionally, in silico toxicity assessment of the detected transformation products of the analyzed substances was also evaluated.

Photodegradation Study of Sertindole by UHPLC-ESI-Q-TOF and Influence of Some Metal Oxide Excipients on the Degradation Process.

The evaluation of the influence of the excipients present in the pharmaceutical formulations on the drug stability is an important part of quality control of medicines. One of the most commonly applied group of excipients are pigments, such as titanium dioxide or various forms of iron oxides, which are well-known photocatalytic agents. Therefore, the photostability of an atypical antipsychotic drug sertindole and the influence of pigments commonly used in the pharmaceutical formulations (FeOOH, Fe2O3, and TiO2) on this process were studied. The quantitative and qualitative analysis of the process was performed with the use of ultra high pressure liquid chromatography with diode array detection (UHPLC-DAD) system coupled with a high resolution hybrid electrospray ionization quadrupole time-of-flight (ESI-Q-TOF) mass spectrometer. Sertindole turned out to be a highly photolabile molecule. Overall 18 transformation products were found, mainly formed as a consequence of dechlorination, hydroxylation, and dehydrogenation. In all the experiments, except the TiO2-mediated photocatalysis, the product of chlorine substitution with a hydroxyl group was the major product. The presence of Fe2O3 and TiO2 accelerated the degradation process, while FeOOH served as its inhibitor. The experiments conducted with the use of the pharmaceutical formulations confirmed the catalytic activity of the used excipients. The exploration of the obtained phototransformation profiles with the use of principal component analysis (PCA) revealed that the presence of both iron oxides could influence the qualitative and quantitative aspect of the studied processes. In silico assessment of the properties showed that the transformation products are generally less toxic to rodents, possess lower hERG blocking potential, but could be more mutagenic than the parent molecule.

Multivariate comparison of photocatalytic properties of thirteen nanostructured metal oxides for water purification.

Although the environmental photocatalysis is being developed for many years, the relationships between simple metal oxides have not been explored so far. In this study a multivariate comparison of thirteen nanostructured metal oxides (Bi2O3, CeO2, Co3O4, Fe2O3, NiO, Pr6O11, SnO2, SrTiO3, TiO2, WO3, ZnFe2O4, ZnO and ZrO2) was performed. The solution containing twenty-six psychotropic pharmaceuticals was used as a model mixture. In order to ensure the influence of the dissolved organic matter on the process, all the experiments were conducted in the river water. Simulated solar radiation was applied as the most environmentally relevant. The high-resolution LC-MS profiles, obtained for the photocatalytic samples after 1 h of irradiation, were then submitted to the multivariate chemometric analysis. Graphical representations of principal component analysis and hierarchical cluster analysis enabled visualization of the relationships between the studied oxides. The registered degradation profiles were compared qualitatively and discussed.

Rapid degradation of clozapine by heterogeneous photocatalysis. Comparison with direct photolysis, kinetics, identification of transformation products and scavenger study.

In this study TiO2-mediated photocatalytic degradation of the persistent drug clozapine under the simulated solar radiation was studied for the first time. The experiments were conducted both in the ultrapure and river water, which enabled the assessment of the organic matrix impact. The direct and indirect photolysis experiments were conducted for a comparison. Influence of the catalyst loading on the efficiency of the process was also assessed, and the highest catalyst loading (300 mg L-1) was found to be the most effective. The TiO2 photocatalysis was extremely effective for clozapine degradation - the decomposition was almost 300 times faster in comparison to the direct photolysis (t1/2 = 1.7 min, neither clozapine, nor the intermediates were detected after 20 min of irradiation), and presence of the organic matrix did not negatively affect the process. Nevertheless the photocatalytic process turned out to be highly sensitive to act of the ROS scavengers. Thirteen transformation products (TPs) were found and their structures were elucidated by the means of high resolution mass spectrometry. Properties - toxicity, biodegradability, BCF and BAF - of TPs and the parent molecule were estimated with the use of computational methods. Identified TPs were found as generally less toxic and more biodegradable than clozapine.

Photolytic and photocatalytic transformation of an antipsychotic drug asenapine: Comparison of kinetics, identification of transformation products, and in silico estimation of their properties.

The photolytic and photocatalytic transformation of an antipsychotic drug asenapine with the use of H2O2 and TiO2 was studied. A method employing irradiation with a simulated full solar spectrum in the photostability chamber was applied, then the reverse-phase ultra high performance liquid chromatography with diode array detector, coupled with electrospray quadrupole time-of-flight mass spectrometer (RP-UHPLC-DAD - ESI-Q-TOF) was used for the quantitative and qualitative analysis of the processes. The developed quantitative method was fully validated, according to the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines, and the kinetic parameters of asenapine photodecomposition were compared. Nineteen phototransformation products were detected, and their probable structures - mainly hydroxylated and oxidized asenapine derivatives - were suggested. On the basis of the elucidated structures the computational prediction of their toxicity at the various endpoints, as well as bioconcentration factors and biodegradability was performed. The obtained results were then subjected to the principal component analysis (PCA). This chemometric technique facilitated comparison of the applied models, calculated properties of the TPs, and enabled visualization of relationships between them.

Characterization of forced degradation products of toloxatone by LC-ESI-MS/MS.

Forced degradation of toloxatone in solutions under basic, acidic, neutral, photo UV-VIS, photo UVC and oxidative stress conditions was investigated and structural elucidation of its degradation products was performed with the use of UHPLC system coupled ESI-Q-TOF mass spectrometer. Eight degradation products were found and their masses and formulas were obtained with high accuracy (0.09-3.79 ppm). The structure of unknown degradation products were elucidated from MS/MS fragmentation spectra of all analyzed compounds. Additionally, whole signals of decomposed substances were compared chemometrically. It was found that toloxatone is fragile towards basic hydrolysis, oxidative conditions and UVC irradiation. Finally, the toxicity of transformation products was computationally evaluated and compared in multivariate manner.

Investigation of the photolysis and TiO2, SrTiO3, H2O2-mediated photocatalysis of an antipsychotic drug loxapine - Evaluation of kinetics, identification of photoproducts, and in silico estimation of properties.

The photolytic and photocatalytic transformation of loxapine with the use of H2O2, TiO2 and SrTiO3 under the simulated solar radiation was studied. A micro-scale method for simultaneous irradiation of multiple samples in photostability chamber was applied. RP-UHPLC-DAD coupled with ESI-Q-TOF mass spectrometer was used for the quantitative and qualitative analysis of the processes. Influence of catalysts concentration on kinetic parameters of loxapine photodecomposition was evaluated, and TiO2 at medium concentration (100 mg L-1) turned out to be the most effective. Sixteen transformation products were detected and their structures were elucidated. On the basis of the elucidated structures, computational evaluation of toxicity, bioconcentration and bioaccumulation factors as well as biodegradability of transformation products were conducted. The multivariate chemometric method (principal component analysis) was used to compare the calculated properties as well as the applied methods. Most of the transformation products were generally less toxic and more biodegradable than the parent compound.