Spectrophotometric Method Research Articles

Selected micro- and macro-element associations with oxidative status markers in common carp (Cyprinus carpio) blood serum and ejaculate: a correlation study

ABSTRACT The aim of this study was to (1) determine complex interactions between macro- and micro-elements present in blood serum and ejaculate of common carp (Cyprinus carpio), and (2) examine the association between alterations in these macro- and micro-elements with markers of oxidative stress. Blood and ejaculate from 10 male carp were collected in the summer period on the experimental pond in Kolíňany (West Slovak Lowland). Reactive oxygen species (ROS), total antioxidant capacity (TAC), protein carbonyls (PC), and malondialdehyde (MDA) levels were measured in blood serum and ejaculate using spectrophotometric methods. The amounts of elements (Ag, Al, Ba, Co, Li, Mo, Ca, K, Na, and Mg) in all samples were quantified using inductively coupled plasma optical emission spectrophotometry. Data demonstrated significant differences in elemental concentrations between blood and ejaculate, specifically significantly higher ejaculate levels were detected for Ag, Al, Ba, Co, Li, Mo, K, and Mg. Potassium was the most abundant macro-element in the ejaculate, while sodium was the most abundant in blood serum. Among the micro-elements, Al was predominant in both types of samples. It is noteworthy that oxidative status markers including ROS, TAC, and MDA were significantly higher in ejaculate indicating the presence of oxidative stress in C. carpio reproductive tissue. The positive correlations between Mg and Ca in blood serum and ejaculate suggest these elements play a functional role in metabolic and physiological processes. In contrast, the positive correlations of Ba and Al with markers of oxidative stress indicated the association of these metals with induction of oxidative stress. Our findings provide insights into the association of metals with biomarkers of physiological function as well as adverse effects in C. carpio.

Intelligent spectrophotometric resolution platforms for the challenging spectra of ipratropium and fenoterol in their combination inhaler with ecological friendliness assessment.

Asthma and chronic obstructive pulmonary disease (COPD) are the most common diagnoses for adults and children with respiratory tract inflammation. Recently, a novel fixed dose combination consisting of Ipratropium and Fenoterol has been released for the management and control of the symptoms of such disorders. The current research has newly developed andoptimizedthree smart, accurate,simple, cost-effective, and eco-friendly spectrophotometric methods that enabled the simultaneous determination of the drugs under study in their combined inhaler dosage form, without the need for any previous separation steps, using water as a green solvent. The strategy employed was based oncalculating oneor two factors as a numerical spectrum or constant, whichprovided thecomplete removal of any component in the mixture that might overlap and the mathematical filtration of the targeted analyte. The methods developed could be classified into twotypesof spectrophotometric windows. Window I; involved absorption spectrum in their original zero-order forms (°D), which included recently designed methods named induced concentration subtraction (ICS) and induced dual wavelength (IDW). While window III focused on the ratio spectrum as theinduced amplitude modulation (IAM) method. The extremely low absorptivity and lack of distinct absorption maximum in the zero-order absorption spectrum of Ipratropium were two intrinsic challenges that were better overcome by the proposed spectrophotometric methods than by the conventionally used ones. According to ICH guidelines, the proposed methods were validated using unified regression over range 2.0-40.0 µg/mL in the ICS method, while the linearity ranges for the IDW and IAM methods were 5.0-40.0 µg/mL of Ipratropium and 2.0-40.0 µg/mL of Fenoterol. Moreover, the three proposed methods were effectively used to assay the co-formulated marketed inhaler and further expanded to confirm the delivered dose uniformityin compliance with the USP guidelines. Finally, the established methods were evaluated for their greenness and blueness, in comparison to the official and reported analysis methods, using advanced cutting edge software metrics. Furthermore, the suggested techniques adhered well to the white analytical chemistry postulates that were recently published.

A green approach: Simultaneous spectrophotometric detection of Co-administered levamisole and triclabendazole in Fasciola-infected sheep with environmental sustainability assessment

The goal of this work is to evaluate the greenness of six spectrophotometric approaches that have been presented for determining the binary combination of levamisole (LVA) and triclabendazole (TCZ). To evaluate the suggested approaches, the analytical community has taken an interest in using green analytical chemistry practices. Two tools, called "Analytical GREEness" (AGREE) and "Green Analytical Procedure Index" (GAPI), were utilized to assess the degree of greenness. The zero spectrum of TCZ and LVA is compared, and it is seen that their spectra overlap. This means that the six spectrophotometric methods can be used to find them: zero order (D0), first derivative (D1), second derivative (D2), the area under the curve (AUC), mean centering, and bivariate. Method I: The zero-order method (D0) is used to measure TCZ directly at 306 nm because LVA has no absorption. Method II: the first derivative method (D1) was used at wavelengths of 312 nm and 220 nm for TCZ and LVA respectively, and the linear range for TCZ and LVA was 2–18 μg/mL and 2–14 μg/mL, respectively. Method III: For the TCZ, the second derivative method (D2) was used at a wavelength of 312 nm and a linear range of 1–18 μg/mL. Method IV: The area under the curve (AUC) method was used with wavelengths of 202–207 nm and 218–226 nm, and concentrations of 2–14 μg/mL for LVA and 1–9 μg/mL for TCZ. Method V: we used a mean-centering method that involved dividing the mean-centered ratio spectrum by the spectrum of the 6 μg/mL TCZ and finding the mean-centered ratio values at 241 nm for LVA. Method VI: The bivariate method is based on measuring two components at two chosen wavelengths using the Kaiser Method. The wavelengths 215 and 225 nm gave the best value of K, so they could be used to find the concentrations of LVA and TCZ. It ranged from 2 to 16 μg/mL for TCZ and from 2 to 14 μg/mL for LVA. These methods were developed for analyzing binary mixtures in pharmaceutical dosage form and bulk powders with high recoveries. Following ICH guidelines, the developed techniques underwent validation. The results show that there were no significant differences when all methods were compared statistically to the described method for TCZ and LVA.

A tomato a day keeps the beetle away - the impact of Solanaceae glycoalkaloids on energy management in the mealworm Tenebrio molitor.

Solanine (SOL), chaconine (CHA), and tomatine (TOM) are plant secondary metabolites produced mainly by the species of Solanaceae family, such as tomato Solanum lycopersicum L. These glycoalkaloids (GAs) have a wide range of biological activity, also in insects. However, their mechanisms of action are not precisely understood. The purpose of the study was to investigate how pure GAs and tomato leaf extract (EXT) affect glycolysis, Krebs cycle and β-oxidation of fatty acid pathways in Tenebrio molitor L. beetle. For this purpose, the larvae were injected with SOL, CHA, TOM, and EXT at two concentrations (10-8 and 10-5M). For experiments, fat body, gut, and heamolymph samples were collected 2 and 24h after injection. Then, the changes in the expression level of phosphofructokinase, citrate synthase, and β-hydroxyacyl-CoA dehydrogenase were measured using the RT-qPCR technique. The catalytic activity of these enzymes and the carbohydrate level in insects after GA treatment were determined by spectrophotometric method. Furthermore, the analysis of the amount of amino acids in tissues was performed with a GC-MS technique. The results obtained show that the GAs changed the activity and expression of the genes encoding key enzymes of crucial metabolic pathways. The effect depends on the type of GA compound, the tissue tested, and the incubation time after treatment. Furthermore, TOM and EXT affected trehalose concentration in the insect hemolymph and led to accumulation of amino acids in the fat body. The observed changes may indicate a protein degradation and/or enhanced catabolism reactions for the production of ATP used in detoxification processes. These results suggest that GAs alter energy metabolism in the mealworm T. molitor. The study contributes to our understanding of the mechanisms of action of secondary metabolites of plants in insects. This knowledge may allow the design of new natural biopesticides against insect pests because proper energy metabolism is necessary for the survival of the organism.

Sorption of non‐ferrous metal ions from aqueous solutions by polymer compositions based on polyacrylic acid and bentonite clay

AbstractComposite hydrogels (CH) were obtained by copolymerization of acrylic acid with N,N‐methylene‐bis‐acrylamide in the suspension of bentonite clay (BC) by the method “in situ.” At this CH of cross‐linking structure was formed in which, in the space between parts of polymer hydrogels (PH) on the base of polyacrylic acid (PAAc), were introduced particles of mineral. The morphology and structure of CH have been investigated by scanning electron microscopy, which has shown that particles of BC are distributed throughout the whole volume of the composite. The sorption of Cu2+, Ni2+, and Cr3+ ions from aqueous solutions of their salts by sorbents using the spectrophotometric method was studied. It has been determined that the introduction of BC particles into the composition of PH leads to a noticeable increase in their sorption capacity. Determination of the elemental analysis of the composition of sorbents after sorption by X‐ray fluorescence analysis showed that the sorption of non‐ferrous metal ions increases in the row Cr3+ < Ni2+ < Cu2+.

Spectrophotometric Assessment of Pyrocatechol Drug Employing Oxidative Coupling Reaction with 4-amino antipyrine as a Model of Pharmaceutical Compounds .

A rapid, simple, sensitive, and higher stability spectrophotometric method was developed the determine that it is stable Pyrocatechol drug in pure and pharmaceutical preparations. The method is based on an oxidative coupling reaction among Pyrocatechol drug and 4-amino antipyrine (4AAP) in the presence of potassium iodate (KIO3) in an acid medium to form a color complex that stable violet dye and a maximum absorption at 505 nm. The best optimum conditions and other analytical factors have been evaluated. A calibration curve of absorbance of several concentrations that obey the law of Beer-Lambert over the range concentration of pyrocatechol drug 10 to 100 mg/L, with (LoD =1.2*10-4 mg/L), (LoQ=3.4*10-4 mg/L), The relative standard deviations RSD% of the proposed methods were less than 0.68%). In contrast, recovery average and determination parameters are 99.9%. The method was used successfully for the determination of drugs in a pharmaceutical preparation with the best accuracy and precision. At range about 99.1 to 100.2% with RSD less than 0.3%. The analytical result for the estimation of the pure drug is presented together with the use of the proposed process for the analysis of some commercial pharmaceuticals. The data compare favorably with those of official and reported ways.

Effects of Aqueous Extracts of Costus afer and Vit. C Supplement on Renal Damage in Male Mice Subjected to Sub-chronic Exposure of Dichlorvos

Aim: To evaluate effects of aqueous extracts of Costus afer and Vit. C supplement on renal damage in male mice subjected to sub-chronic exposure of dichlorvos. Study Design: The study was a completely randomized design employing relevant statistical tools for analysis and interpretation. Place and Duration of Study: The study was carried out in the Department of Animal and Environmental Biology, Rivers State University. The experiment lasted for 35 days between April and June 2023. Methodology: A total of 30 male mice were randomly selected into 6 groups (n=5). Group A was the control, B received Dichlorvos@25mg/kg/bw/day, C & D received 100% vitamin C and Costus afer at 250mg/kg/bw/day only. Groups E & F were coadministered Dichlorvos with Vitamin C and 100% extract of Costusaferat 250mg/kg/bw/day respectively. At the end of the treatment period, blood samples were collected for analysis. Total protein was analyzed using the spectrophotometric method of biuret, Bradford and erythrosine – b, albumin was estimated, creatinine and urea was done using enzymatic method.Histological sections of the kidney were mounted on slides, stained with hematoxylin and eosin (H&E). Photomicrographs were generated. Results: All kidney biomarkers examined increased significantly (p=0.05) in group B administered Dichlorvos only but decreased in groups administered the vit C and Costusafer respectively. Histopathological analysis of the kidney shows the regular structure of the kidney architecture in group A. Glomeruli shrinkage, necrosis and degeneration of nephrons were observed in group B, while less pronounced changes were observed in groups C-F co-administered with Vit C and Costus afer. However, group E shows fully regenerated nephrons while few depleted nephronic cells were still observed in group F despite coadministration of Costus afer. Conclusion: Based on the results, Vit C supplement shows more potent, therapeutic properties against kidney damage in male mice subjected to sub-chronic exposure of Dichlorvos compared toCostusafer.

Analysis of chemical composition, antimicrobial, antioxidant activities of pomegranate fruit thick and green tea leaf extracts

Infection diseases is a worldwide important problem for medicine and pharmacy. The purpose of this work was to study and compare phytochemical composition, antimicrobial, antioxidant potential of pomegranate fruit thick and green tea leaf liquid extracts. The quantification of biologically active substances (BAS) was performed using spectrophotometric, titrimetric, and HPLC analysis methods. Antioxidant activity was measured through a potentiometric method, while antimicrobial and antifungal effects were assessed using the well method and determining the minimum inhibition concentration. The total content of phenolic compounds was 0.40 and 10.10%, organic acids – 5.80 and 1.60% for pomegranate fruit thick and green tea leaf extract. The total content of catechins in the green tea leaf extract was 10500.0 mg/100 g, where epicatechin-3-O-gallate was dominated (3730.0±2.00 mg/100 g). The total content of anthocyanins in the pomegranate fruit thick extract was 107.10 mg/100 g, where cyanidin-3-O-glucoside was dominated (41.74 ± 0.10 mg/100 g), whereas delphinidin-3-O-glucoside had a lowest content (4.99 ± 0.10 mg/100 g). Both extracts possessed a high antioxidant potential, and effective antimicrobial effects. The antioxidant, antimicrobial activity of pomegranate fruit extract was higher than green tea leaf extract. In addition, we assumed that anthocyanins had higher antioxidant, antimicrobial properties than catechins. These findings would promote application of pomegranate fruits extract as pharmaceuticals and nutraceuticals.

Potential benefit of Lactobacillus acidophilus supplementation to rats fed with a high-fat diet on serum lipid profile, kidney amyloid protein and tumor necrosis factor-alpha level

AimIt was aimed to determine the potential effect of Lactobacillus acidophilus supplementation on rats exposed to an experimental high-fat diet on serum lipid profile and kidney total beta amyloid protein (TBAP) and Tumor Necrosis Factor-alpha (TNF-α) levels. Methods24 male Sprague-Dawley rats were used in the study to establish 4 groups. Standard rat food (SD) was provided to Group 1 as the control; Group 2 was fed a high-fat diet (HFD); Group 3 consumed SD and received L. acidophilus probiotics; Group 4 was fed HFD and received L. acidophilus probiotics. Body weights were determined weekly during the 12-week trial period. At the end of the experiment, TBP and TNF-α levels in the serum and kidney tissue of the rats were measured by ELISA method. Serum total cholesterol (TC), triglyceride (TG), HDL, LDL, urea and creatinine levels and paraoxanase, amylase and lipase activities were determined by spectrophotometric method on the analyzer device. ResultsWhen the control (Group 1) group and Group 2 were compared at the end of the experiment, it was found that Group 2 had gained the most weight and that both the blood and kidney tissue levels of TNF-α and TBAP, as well as the quantities of TG, TK, LDL, and urea, were significantly greater (P<0.05). Serum HDL, PON and amylase levels were found to be significantly low (P>0.05). TG, TK, LDL, urea, and the levels of TNF-α and TBAP in serum and renal tissue were shown to be lower in the groups who received L. acidophilus probiotics (Groups 3, 4) when compared to Group 2 (P>0.05). It was observed that HDL, PON and amylase levels increased and approached the control group (P<0.05). ConclusionThe study's findings showed that probiotic supplementation improved blood levels of TG, TC, HDL, LDL, urea, PON, and amylase as well as serum and kidney tissue levels of TNF-α and TBAP in obese rats fed a high-fat diet.

Chemical Derivatization and Paper Spray Ionization Mass Spectrometry for Fast Screening of Retinoic Acid in Cosmetics.

As a prescription drug, retinoic acid is listed as a banned cosmetic additive in the EU and China regulations. Currently, spectrophotometric methods, including thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and HPLC-MS/MS, are commonly used for the determination of retinoic acid. As these conventional methods require complex pretreatment and are time-consuming, chemical derivatization combined with paper spray ionization mass spectrometry was developed for the fast detection of retinoic acid in cosmetics. N,N-dimethylpiperazine iodide (DMPI) was utilized as a derivatization reagent. Carboxylic acid in retinoic acid was derivatized to carry a positive charge and was subjected to mass spectrometry analysis. Results showed that compared with non-derivatized compounds, the detection limit was increased by about 50 times. The linearity in the range of 0.005-1 μg·mL-1 was good. The limit of detection (LOD) was 0.0013 μg·mL-1, and the limit of quantification (LOQ) was 0.0043 μg·mL-1. The recoveries of spiked samples were in the range of 95-105%, and the RSDs were below 5%. Derivatization and paper spray ionization MS render a quick, sensitive, and accurate method for the detection of retinoic acid in a complex matrix.

Phytochemical Analysis and Nanoparticle Formulations of Extracts Myristica fragrans Houtt Leaves as Antibacterial

Myristica fragrans Houtt leaves contain secondary metabolites such as flavonoids, terpenoids and alkaloids, which have antibacterial properties. At the nanoscale, the particles have a bigger surface contact area, resulting in increased amounts and solubility of the active chemical. This leads to a stronger antibacterial activity. This study aims to examine the influence of several chitosan variations on particle size, antibacterial properties and the form of nanoparticles produced from Myristica fragrans leaves ethanol extract. Ethanol extract was analyzed using the UV-Vis spectrophotometric method to quantify total flavonoid content. Nanoparticles were synthesized using the ionic gelation technique with several ratios of chitosan and sodium tripolyphosphate, specifically 8:1, 10:1 and 12:1. Nanoparticles were analyzed with a Particle Size Analyzer (PSA) and Scanning Electron Microscopy (SEM). Agar diffusion test for bacteria germs using a paper backer. Myristica fragrans leaves ethanol extract was 50, 40 and 30 %, while nanoparticle extract was 5, 4 and 3 %. The overall flavonoid concentration in the QE/ethanol extract was 39.7252 ± 1.9596 mg. Antibacterial test results against Staphylococcus aureus and Escherichia coli bacteria and Myristica fragrans leaves extract limit inhibition area at 30 % (13.35 mm), 40 % (13.68 mm) and 50 % (13.93 mm) for Staphylococcus aureus and 30 (13.28), 40 (13.45) and 50 % (13.81 mm) for Escherichia coli. Myristica fragrans leaves extract chitosan nanoparticles included 3 (14.70), 4 (15.30) and 5 % (15.56 mm) for Staphylococcus aureus and 3 (14.60), 4 (14.65) and 5 % (15.05 mm) for Escherichia coli. Ionic gelation can create nanoparticles of Myristica fragrans leaves ethanol extract with chitosan and sodium tripolyphosphate. Myristica fragrans leaves nanoparticle ethanol extract exhibits better antibacterial activity than ethanol extract. Increasing chitosan concentration affects particle size. The ethanol extract of Myristica fragrans nanoparticle leaves showed slightly better antibacterial activity than the ethanol extract. HIGHLIGHTS Nanoparticles of extracts and extracts have proven excellent antibacterial activity against human pathogens through paper disc diffusion method. Antibacterial test results against Staphylococcus aureus and Escherichia coli bacteria and Myristica fragrans leaves extract limit inhibition area at 30 % (13.35 mm), 40 % (13.68 mm), and 50 % (13.93 mm) for Staphylococcus aureus and 30 % (13.28 mm), 40 % (13.45 mm), and 50 % (13.81 mm) for Escherichia coli. Myristica fragrans leaves extract chitosan nanoparticles included 3 % (14.70 mm), 4 % (15.30 mm), and 5 % (15.56 mm) for Staphylococcus aureus and 3 % (14.60 mm), 4 % (14.65 mm), and 5 % (15.05 mm) for Escherichia coli. The synthesized extract nanoparticles can be used against human pathogens acts as antibacterial drugs. GRAPHICAL ABSTRACT

Sustainable and technically smart spectrophotometric determination of PAXLOVID: a comprehensive ecological and analytical performance rating

The Food and Drug Administration (FDA) authorized the administration of ritonavir (RIT)-boosted nirmatrelvir (NMV) on May 25, 2023, for the treatment of mild to moderate COVID-19 in patients who are at high risk of developing severe COVID-19. In accordance with sustainability and environmental friendliness, simple, eco-friendly, and sustainable spectrophotometric methods were established for concurrently estimating RIT and NMV in newly launched copackaged pills. The suggested solutions for resolving the spectral overlap between RIT and NMV involve the following mathematical methods: the first derivative method (1D), second derivative method (2D), and dual-wavelength zero-order method (DWZ). When ethanol was used as a green dilution solvent, the linearity range was adjusted (10–250 µg/mL) for both drugs. The procedures resulted in a high correlation coefficient (not less than 0.9996) and satisfactory levels of detection and quantification. Additionally, method validation was performed in accordance with International Council for Harmonization norms. Moreover, a detailed ecological and sustainability evaluation protocol was established to confirm the greenness and whiteness of the methods. Finally, the proposed method, along with previously reported methods for analysing NMV and RIT, were reviewed analytically and ecologically.

Exploring antioxidative properties of xanthohumol and isoxanthohumol: An integrated experimental and computational approach with isoxanthohumol pKa determination

This study explores the antioxidative activities of xanthohumol (XN) and isoxanthohumol (IXN), prenylated flavonoids from Humulus lupulus (family Cannabaceae), utilizing the oxygen radical absorption capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays along with computational Density Functional Theory methods. Experimentally, XN demonstrated significantly higher antioxidative capacities than IXN. Moreover, we determined IXN pKa values using the UV/Vis spectrophotometric method for the first time, facilitating its accurate computational modeling under physiological conditions. Through a thermodynamic approach, XN was found to efficiently scavenge HOO• and CH3O• radicals via Hydrogen Atom Transfer (HAT) and Radical Adduct Formation (RAF) mechanisms, while CH3OO• scavenging was feasible only through the HAT pathway. IXN exhibited its best antioxidative activity against CH3O• via both HAT and RAF mechanisms and could also scavenge HOO• through RAF. Both Single Electron Transfer (SET) and Sequential Proton Loss-Electron Transfer (SPLET) mechanisms were thermodynamically unfavorable for all radicals and both compounds.

In vitro and in silico therapeutic properties of Artepillin C and Aromadendrin as collagenase and elastase inhibitors and investigation of anti-Ovarian cancer effects and antioxidant potential

In this investigation, the enzymes collagenase and elastase were inhibited by artepillin.C and aromadendrin molecules with excellent to good IC50 values of 16.65, and 0.79 μM for artepillin.C and 7.31, 19.38, and 10.56 μM for aromadendrin. Using the molecular modeling study, the chemical activity of these compounds against the enzymes, collagenase, and elastase were evaluated. The anti-cancer properties of the compounds were evaluated using the following cell lines: NIH: OVCAR-3, ES-2, UACC-1598, Hs 832(C).T [Hs832.Tc], TOV-21G, UWB1.289. The DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical scavenging assay was used to measure antioxidant activity, and the spectrophotometric method was used in this work. The chemical activities of artepillin.C and aromadendrin against collagenase and elastase were investigated utilizing the molecular docking study. The anti-cancer activities of the compounds were evaluated against NIH: OVCAR-3, ES-2, UACC-1598, Hs 832(C).T [Hs832.Tc], TOV-21G, UWB1.289 cell lines. Also, some cell lines had best results for aromadendrin like NIH: OVCAR-3, ES-2, UACC-1598, Hs 832(C).T [Hs832.Tc], TOV-21G, UWB1.289 (10.54 ± 0.94, 22.11 ± 2.52, 31.85 ± 4.73, 8.14 ± 1.52, 17.94 ± 1.88, and 24.31 ± 2.64 μM). The chemical activities of artepillin.C and aromadendrin against some of the expressed surface receptor proteins (folate receptor, CD44, EGFR, Formyl Peptide Receptor–Like 1, M2 muscarinic receptor, and estrogen receptors) in the mentioned cell lines were assessed using the molecular docking calculations. The outcomes provided information on potential interactions and their atomic-level properties. According to the docking scores, the compounds show a high affinity for binding to proteins and enzymes. Furthermore, these substances made good contact with the receptors and enzymes. As a result, these substances may have the ability to suppress cancer cells and enzymes.

Association of antibiotic resistance and biofilm formation in Escherichia coli ST131/O25b.

Urinary tract infections are becoming difficult to treat every year due to antibiotic resistance. Uropathogenic Escherichia coli (UPEC) isolates pose a threat with a combined expression of multidrug-resistance and biofilm formation. ST131 clone is a high-risk pandemic clone due to its strong association with antimicrobial resistance, which has been reported frequently in recent years. This study aims to define risk factors, clinical outcomes, and bacterial genetics associated with ST131/O25b UPEC. In this study, antibiotic susceptibility and species-level identification of 61 clinical E. coli strains were determined by automated systems. Detection of extended-spectrum beta-lactamases was assessed by double-disk synergy test. Biofilm formation was quantified by spectrophotometric method. Virulence genes (iutA, sfa cnf-1, iroN, afa, papA, fimA), antibiotic resistance genes (blaCTX-M, blaTEM, blaSHV, blaOXA, qnrA, qnrB, qnrS, ant(2')-Ia, ant(3)-Ia, aac(3)-IIa, mcr-1, mcr-2, mcr-3, mcr-4) were investigated by PCR. The following beta-lactamase genes were identified, blaTEM (n = 53, 86.8%), blaCTX-M (n = 59, 96.7%), blaSHV (n = 47, 77.0%), and blaOXA-1 (n = 27, 44.2%). Our data revealed that 93.4% of (57/61) E. coli isolates were biofilm-producers. O25pabBspe and trpA2 were investigated for the presence of ST131/O25b clone. Among multidrug resistant isolates, co-existence of O25pabBspe and trpA2 was detected in 29 isolates (47.5%). The fimH30 and H30Rx subclones were detected in four isolates that are strong biofilm-producers. These results suggest that clinical E. coli strains may become reservoirs of virulence and antibiotic resistance genes. This study demonstrates a significant difference in biofilm formation between E. coli ST131 and non-ST131 isolates. Moreover, 86.21% (n = 25) of ST131 isolates produced strong to moderate biofilms, while only 43.75% (n = 14) of non-ST131 isolates showed the ability to form strong biofilms. Presence of iutA and fimA genes in the majority of ST131 strains showed an important role in biofilm formation. These findings suggest application of iutA and fimA gene suppressors in treatment of infections caused by biofilm-producing drug-resistant ST131 strains.

Analytical Methods for Assessing Thiol Antioxidants in Biological Fluids: A Review.

Redox metabolism is an integral part of the glutathione system, encompassing reduced and oxidized glutathione, hydrogen peroxide, and associated enzymes. This core process orchestrates a network of thiol antioxidants like thioredoxins and peroxiredoxins, alongside critical thiol-containing proteins such as mercaptoalbumin. Modifications to thiol-containing proteins, including oxidation and glutathionylation, regulate cellular signaling influencing gene activities in inflammation and carcinogenesis. Analyzing thiol antioxidants, especially glutathione, in biological fluids offers insights into pathological conditions. This review discusses the analytical methods for biothiol determination, mainly in blood plasma. The study includes all key methodological aspects of spectroscopy, chromatography, electrochemistry, and mass spectrometry, highlighting their principles, benefits, limitations, and recent advancements that were not included in previously published reviews. Sample preparation and factors affecting thiol antioxidant measurements are discussed. The review reveals that the choice of analytical procedures should be based on the specific requirements of the research. Spectrophotometric methods are simple and cost-effective but may need more specificity. Chromatographic techniques have excellent separation capabilities but require longer analysis times. Electrochemical methods enable real-time monitoring but have disadvantages such as interference. Mass spectrometry-based approaches have high sensitivity and selectivity but require sophisticated instrumentation. Combining multiple techniques can provide comprehensive information on thiol antioxidant levels in biological fluids, enabling clearer insights into their roles in health and disease. This review covers the time span from 2010 to mid-2024, and the data were obtained from the SciFinder® (ACS), Google Scholar (Google), PubMed®, and ScienceDirect (Scopus) databases through a combination search approach using keywords.

Quantification of morphine in exhaled breath condensate using a double network polymeric hybrid hydrogel functionalized with AuNPs

BackgroundMorphine serves as a foundation for creating other opioid derivatives, such as hydro/oxymorphine and heroin, which possess enhanced pain-relieving properties but are also prone to addiction and abuse. In cases of morphine overdose, it not only affects multiple immune functions but can also cause severe health complications. Given these concerns and the widespread use of morphine, it is crucial to develop efficient, uncomplicated, and precise methods for accurately detecting morphine in various biological and pharmaceutical samples.ResultsIn this investigation, a novel gold nanoparticle (AuNPs)-based double network hydrogel (DNH) nanoprobe has been fabricated for sensitive quantification of morphine in exhaled breath condensate samples. For that, gelatin/agarose DNH was fabricated through a one-step heating-cooling method in the presence of AuNPs, providing not only chemical stability but also prevent the AuNPs aggregation during synthesis process. In this method, the absorbance intensity of the nanoprobe gradually decreased with increasing morphine concentration due to the interaction morphine with AuNPs surface plasmon. The aggregation of AuNPs by addition of morphine was verified by UV-Vis spectrophotometry. The sensor displayed high sensitivity with detection limit of 0.006 µg.mL-1 in the linear range from 0.01 to 1.0 µg.mL-1. A reliable performance was attained for the spectrophotometric method for determination of morphine in the real samples.

Determination of ammonium and nitrate in soils by digital colorimetry.

A method of digital colorimetric determination of ammonium and nitrate in soils is proposed. The method is based on corresponding photometric procedures of ammonium and nitrate determination after potassium chloride extraction from soil samples. Ammonium is determined as an indophenol dye, and nitrate is determined as an azo dye. The original procedures were modified to overcome the lower sensitivity of the digital colorimetric method. For ammonium determination, the time required for the reaction to proceed completely was studied. Along with the use of a 96-well microplate protected from ambient light by a special frame, mathematical correction of scattered radiation using black ink and taking the images by a scanner in transmission mode without any post-processing, the resulting colorimetric methods proved to provide accuracy and sensitivity close to those of the spectrophotometric method, and the overall analysis speed for tens of samples was even higher. Limits of detection and quantitation for NO3- were 0.42 and 1.4mg/kg, and for NH4+, they were 1.1 and 3.7mg/kg, which is lower than for standard methods. The methods' validity was proven by the analysis of standard samples and by the analysis of soil samples collected in several districts of the Moscow region.

Development and validation of a green spectrophotometric method for simultaneous determination of combined pharmaceutical dosage form (paracetamol and caffeine) using chemometrics technique in comparison with HPLC

Development and validation of a green spectrophotometric method for simultaneous determination of combined pharmaceutical dosage form (paracetamol and caffeine) using chemometrics technique in comparison with HPLC

Development of an affordable light emitting diode spectrophotometer paired with a Python program for calibration and linearity testing and the measurement of uranium(VI).

Uranium (U) is a radiologically and chemically toxic element that occurs naturally in water, soil, and rock at generally low levels. However, anthropogenic uranium can also leach into groundwater sources due to mining, ore refining, and improper nuclear waste management. Over the last few decades, various methods for measuring uranium have emerged; however, most of these techniques require skilled scientists to run samples on expensive instrumentation for detection or require the pretreatment of samples in complex procedures. In this work, a Schiff base ligand (P1) is used to develop a simple spectrophotometric method for measuring the concentration of uranium (VI) with an accurate and affordable light-emitting diode (LED) spectrophotometer. A test for a higher-order polynomial relationship was used to objectively determine the calibration data's linearity. This test was done with a Python program on a Raspberry Pi computer that captured the spectrophotometer's calibration and sample measurement data.