Routine Analysis Research Articles

Green spectrophotometric approaches applied to tertiary mixture for management of common cold and COVID-19 symptoms

Recently, cold and cough dosage forms have gained significant attention due to their use in the supportive protocols for managing COVID-19 symptoms. In this study, a pharmaceutical formulation containing Paracetamol (PAR), Guaifenesin (GUA), and Phenylephrine hydrochloride (PHE) was investigated for spectral resolution and quantification using advanced spectrophotometric methods. The spectra of these components were significantly overlapped and present in their combined tablet in a challenging ratio of 250:100:5 for PAR, GUA, and PHE, respectively. The established approaches were employed for the simultaneous determination of these drugs in their pharmaceutical formulation without interference from matrix excipients. The study involved various manipulation steps, allowing each component in the combination to be analyzed by more than one approach. Integrating these methods with smart mathematical techniques, the issue of spectral data overlap was resolved without the need for preliminary separation steps. The developed methods are dual wavelength, first derivative, derivative ratio, ratio difference, constant center coupled with spectrum subtraction, and constant multiplication paired with spectrum subtraction. The proposed methods were linear over the concentration range of 3.0–35.0 μg/mL for GUA and 3.0–30.0 μg/mL for PHE. While the PAR ranges for the first derivative and constant multiplication methods were 2.5–35.0 μg/mL and 2.5–25.0 μg/mL, respectively. Excellent linearity of the suggested methods was demonstrated by the high correlation coefficients (R2), ≥ 0.9998 for all the tested compounds. These methodologies were validated according to ICH guidelines. Validation results demonstrated excellent accuracy, with recovery percentages ranging from 98 to 102 %, and precision, with RSD values less than 2 %. The obtained results were statistically compared with the official ones using F-test, Student’s t-test, and one-way ANOVA, revealing no significant differences. The proposed methods are accurate, green, smart, fast, and cost-effective. Their compliance with Green Analytical Chemistry principles was evaluated and compared to a published method using various tools to enable a more holistic evaluation from different perspectives. The promising results revealed that the investigated methods are superior green alternatives for routine analysis of the cited drugs in laboratories with limited resources and without access to expensive instruments.

Ontology modeling for automation of questionnaire data processing

The object of this study is the analysis of questionnaire data using ontological modeling. The task relates to the fact that conventional methods for processing questionnaire data are often insufficiently effective when working with large volumes of information and do not make it possible to automate many analysis processes. As a result of the study, an ontology was designed that structures and analyzes questionnaire data, which allows for a more accurate identification of hidden relationships between variables. Using these theoretical provisions, an information system for assessing the quality of assimilation of preschool children's competencies was built. 150 children from various preschool organizations were involved in the study as respondents. The data integration method proposed in this paper significantly facilitated the process of data analysis both for a group and for an individual respondent. The key difference of the proposed methodology is the automation of routine data analysis operations based on the ontological structure, which significantly simplifies the processing of large volumes of information. This makes it possible to solve the problem of limitations in conventional analysis methods and makes data analysis more scalable and reproducible. The practical application of the results is possible in marketing for analyzing customer satisfaction, market segmentation, and evaluating the effectiveness of advertising campaigns. In the educational domain, the ontology could be used to evaluate the quality of programs and analyze respondents' opinions, and in sociology – to analyze public opinion and conduct research on social phenomena. Thus, the proposed ontology provides an effective tool for analyzing large volumes of questionnaire data, allowing organizations to make more informed decisions and improve their efficiency

Use of stable isotope combined with intact cell lipidomic by routine MALDI mass spectrometry analysis for rapid drug susceptibility assay in mycobacteria.

Rapid, accurate, and easy-to-perform diagnostic assays are required to address the current need for the diagnosis of resistant pathogens. That is particularly the case for mycobacteria, such as the human pathogen Mycobacterium tuberculosis, which requires up to 2weeks for the determination of the drug susceptibility profile using the conventional broth microdilution method. To address this challenge, we investigated the incorporation of deuterium, the stable isotope of hydrogen, into lipids as a read out of the drug susceptibility profile. Deuterium is incorporated into newly synthesized proteins or lipids in place of hydrogen as bacterial cells grow, increasing the mass of the macromolecules, which can then be observed via matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). As proof-of-concept, we used the non-pathogenic Mycobacterium smegmatis mc2155 strain, which is susceptible to the aminoglycoside antibiotic kanamycin, and M.smegmatis mc2155 containing the empty vector pVV16, which is kanamycin-resistant. Bacteria were incubated in a culture medium containing 50% of deuterium oxide (D2O) and either 1 or 2 times the minimal inhibitory concentration (MIC50) of kanamycin. Lipids were then analyzed using the MBT lipid Xtract matrix combined with routine MALDI mass spectrometry in the positive ion mode to evaluate the changes in the lipid profile. Using this approach, we were able to distinguish susceptible from resistant bacteria in less than 5h, a process that would take 72 h using the conventional broth microdilution method. We therefore propose a solution for the rapid determination of drug susceptibility profiles using a phenotypic assay combining D2O stable isotope labelling and lipid analysis by routine MALDI mass spectrometry.

Quality by Design Approach Driven Development and Validation of Reverse Phase‐High‐Performance Liquid Chromatography Methodology for Dual Quantification of Aspirin and Rivaroxaban

ABSTRACTClinical trials have shown that combining a modest dose of Rivaroxaban (RIV) with Aspirin (ASP) is more effective than ASP alone in reducing cardiovascular mortality, myocardial infarction, and stroke. A high‐performance liquid chromatography method developed using Quality by Design (QbD) effectively estimates ASP and RIV for routine pharmaceutical analysis. The Plackett‐Burman design assessed key factors including organic phase volume, mobile phase pH, flow rate, temperature, and injection volume using an Ishikawa fish‐bone diagram for risk assessment. These parameters were then optimized via Box‐Behnken design in 15 trials. The method meets the International Council For Harmonization standards with a mobile phase ratio of 45:55 (v/v) acetonitrile: buffer (pH 3, adjusted with orthophosphoric acid), a flow rate of 1 mL/min, a column temperature of 30°C, and an injection volume of 20 µL. A Phenomenex Luna C18 column (250 × 4.6 mm, 5 µm particle size) with detection at 251 nm ultra‐violet wavelength. The retention times of 4.54 min for ASP and 5.81 min for RIV were obtained. Validation confirmed linear ranges: 40–200 µg/mL for ASP and 2–10 µg/mL for RIV, with excellent recovery rates (ASP: 99.30%–101.9%; RIV: 98.52%–101.84%). Developed using QbD principles, the method proved specific, accurate, precise, sensitive, and robust for routine quality control of both compounds in pharmaceutical formulations.

Ion-chromatographic determination of common anions in drinking water in some regions of the Republic of Armenia

The present study investigated the anion composition of drinking water from various cities and villages in Armenia, including spring water and tap water samples. The simultaneous separation was achieved for fluoride, chloride, bromide, nitrite, nitrate, sulfate, and phosphate anions with the correlation coefficients >0.998. Validation of the method was carried out in accordance with the requirements of guidelines in terms of selectivity, specificity, system suitability, linearity, accuracy, precision, lower limits of detection and quantitation, robustness, and stability. The method was suitable for routine drinking water quality assessment in line with the national or World Health Organization (WHO) regulations. The validated method was further used to analyze drinking water samples from different regions of Armenia. The observed concentrations for the anions in drinking water complied with the WHO limits for drinking water with some exceptions. The test method can be submitted to quality control laboratories as a convenient and practical tool for routine analysis and monitoring studies.

RGB Trichromatic Whiteness Assessment of Bio Analytical Chromatographic Tool Using Fluorescence for Quantitation of Semaglutide: Application to Pharmaceutical Preparations and Spiked Plasma.

Semaglutide (SEMG) is one of the most widely used and trending medications to treat type II diabetes and obesity.This work aimed to develop a liquid chromatography with spectroflourimetric detection (HPLC-flourimetry) analysis of SEMG in both its tablet dosage form and plasma. The power of fluorescence detection coupled with HPLC proved its capability as a bioanalytical tool to assay SEMG in plasma samples owing to its simplicity and sensitivity which reached below the Cmax of SEMG. Separation was done using a C18 column with mobile phase of acetonitrile and water acidified with orthophosphoric acid (pH 3.5) (1.41 × 10-5M) in isocratic mode in ratio 57:43 and 1mL/min flow rate after extraction using protein precipitation. Detection was carried out at λ excitation of 238nm and λ emission of 416 and 307nm for SEMG and the internal standard, respectively. Evaluation of greenness of the proposed method was done using AGREE (Analytical GREEnness Metric Approach), ComplexGAPI (Complementary Green Analytical Procedure Index) & the new algorithm RGB 12 model (Red-Green-Blue). They showed that these methods can be a greener alternative with acceptable sensitivity for analysis of SEMG. The developed seven min-assay was validated per ICH as well as FDA bio analytical methods' guidelines to prove its applicability for routine sample analysis and future pharmacokinetic studies.

Four ecofriendly spectrophotometric methods for the determination of perindopril through derivatization with sulphophtalein dyes: application to tablet analysis

Nowadays, there is a need to expand the bank of spectrophotometric methods for the determination of perindopril in dosage forms for the purposes of routine pharmaceutical analysis, which would be simple, express, «green» and inexpensive. In the present work, perindopril in tablets was quantified via a direct simple, «green», and non-extracting spectrophotometric approach based on the formation of ion-pair complexes with sulphophtalein dyes. The absorbances of the colored reaction products were registered at 405 nm (bromocresol green, BCG), 397 nm (bromocresol purple, BCP, and bromothymol blue, BTB) and 598 nm (bromophenol blue, BPB). To achieve the highest intensity of absorbance, optimum conditions were established by the screening of many experimental factors such as optimal concentration and volume of dyes, and the time consumed for the reaction. Beer’s law was obeyed in the ranges of 0.44–3.96 µg/mL (BCG), 3.00–7.00 µg/mL (BCP), 4.00–12.00 µg/mL (BTB) and 0.44–3.52 µg/mL (BPB). All four methods were validated in accordance with ICH guidelines, confirming specificity and linearity, accuracy and precision, limits of detection and quantification, robustness. These validated methods provide a reliable and green approach for the quantitative analysis of perindopril in tablets, contributing to safer and more sustainable laboratory practices in pharmaceutical analysis.

3D Printed Metallic Pillar Nanomechanical Resonators Decorated with TiO2 Nanotubes for Highly Sensitive Environmental Applications

AbstractMicro and nanomechanical devices offer enhanced sensing capabilities for detecting biological and chemical small molecules. However, miniaturization necessitates advanced fabrication processes and complex measurement systems, hindering routine sensor analysis. While alternative methods like 3D printing show promise, challenges such as low device resolution persist due to intrinsic damping of polymer inks. In this study, an array of micrometric pillar resonators is fabricated in Ti6Al4 V alloy using additive manufacturing based on laser powder bed fusion technology. These metallic nanomechanical resonators exhibit a very high quality factor with minimal difference between air and vacuum measurements due to low intrinsic damping. Furthermore, titania nanotubes grown on the pillars via anodic oxidation heighten sensitivity for molecular dye degradation evaluation. Leveraging the weak coupling phenomenon among the pillars in the array, these devices facilitate large‐scale parallelized measurements, here demonstrated with real‐time analysis of dye degradation process. This approach to creating mass sensing devices via metallic additive manufacturing can usher in a new generation of highly performing resonating sensor arrays, offering a cost‐effective and efficient alternative to traditional silicon microfabrication methods.

Application ICP-OES to Multielement Analysis on Plastic Waste and Blends with Vacuum Gas Oil: Developing a Sample Preparation Protocol

This paper introduces a new methodology for a routine metal analysis of plastic waste (PW) and PW blended with petroleum feedstock such as vacuum gas oil and VGO (PW/VGO). For such purposes, recycled polyethylene and polypropylene plastic were selected to mimic the potential feeds to be integrated at the Fluid Catalytic Cracking unit (FCC) to produce valuable products. Elements such as P, Ca, Al, Mg, Na, Zn, B, Fe, Ti, and Si were included in the method development. Different sample preparation methods were evaluated, such as microwave-assisted acid digestion (MWAD) and dry/wet ashing, followed by a fusion of the ash with lithium borate flux. Some PW homogenization pretreatments, such as cryogenic grinding and hot press molding, were also covered. The finding of this work suggests that MWAD with HNO3 and H2O2 is adequate for both types of samples and is the quickest sample preparation; however, the sample needed to be homogenized, and recoveries for Si and Ti may be biased for PW due to the limited solubilities of these elements in the nitric acid media. Carbon removal is required before fusion sample preparation and analysis due to the amount of carbon in PW samples. The sample needed to be homogenized for wet ash fusion but not for the pre-ash (dry) method. A benefit to the damp ash pretreatment is that the ash for the sample was created in the same crucible used for fusion digestion, avoiding material loss during sample management. Fusion from wet ash or carbon removal allowed for better acid solubility for Si and Ti in PW. The results of the PW samples evaluated matched well with those of both sample preparation methodologies. For most elements, precision was <10% regardless of the sample preparation; however, Fe and P had some variation using wet ash fusion, possibly due to contamination in an open digestion system or variation due to being close to the method limit of quantification (LOQ). The methodology reported here is robust enough to be implemented as routine analysis in any laboratory facility.

Scientifc Validation and Chromatographic Method Development for the Estimation of Piperine in Herbal Formulation Babbularishta: A Quality Control Strategy for Critical Quality Attributes

Background: Babbularishta is an Ayurvedic medicine in liquid form used for the treatment of phthisis, skin diseases, urinary disorders,, and breathing problems. It is also beneficial in cases of blood disorders, acne, dermatitis, hemoptysis, lung injury, asthma, persistent cough, whooping cough, etc. Objective: The functional role of this study is to develop a simple, novel, precise, accurate, reliable, and selective RP-HPLC method for the estimation of piperine content in herbal drug formulations (Babbularista). Methods: A traditional approach to method development could fail to meet the desired separation. Consideration of the current regulatory requirement for analytical method development and RP-HPLC method for routine analysis of piperine in the herbal formulation has been optimized using an analytical Quality by Design (QbD) approach. Three laboratory formulations and three marketed preparations of Babularishta were investigated for organoleptic parameters, physicochemical parameters, and validation. The mobile phase composition (methanol and water % as A), pH (B), and flow rate (mL/min as C) were selected as independent variable,. In contrast, retention time (min. as Y2), area (AU as Y1) is selected as dependent variables. Results: The fingerprinting method was developed and optimized by using the AQbD approach. Piperine content was found to be in Piper nigrum (Marica) 3.01±0.01527 % w/w and in Piper longum (Pipali) 2.74±0.01154% w/w in Babbularista. Conclusion: An attempt has been made to develop quality control parameters of Babbularista by developing a suitable and sensitive HPLC methods for the quantitation of piperine in herbal drug formulations (Babbularista). The developed method was found to be simple, accurate, precise,, and economical for the estimation of piperine in herbal formulations.

Reconstruction of reservoir rock using attention-based convolutional recurrent neural network

The digital reconstruction of reservoir rock or porous media is important as it enables us to visualize and explore their real internal structures. The reservoir rocks (such as sandstone and carbonate) contain both spatial and temporal characteristics, which pose a big challenge in their characterization through routine core analysis or x-ray microcomputer tomography. While x-ray micro-computed tomography gives us three-dimensional images of the porous media, it is often impossible to quantify the variability of the pore, grains, structure, and orientation experimentally. Recently, machine learning has successfully demonstrated the reconstruction ability of reservoir rock images or any porous media. These reservoir rock images are crucial for the digital characterization of the reservoir. We propose a novel algorithm consisting of the convolutional neural network, an attention mechanism, and a recurrent neural network for the reconstruction of reservoir rock or porous media images. The attention-based convolutional recurrent neural network (ACRNN) can reconstruct a representative sample of reservoir rocks. The reconstructed image quality was checked by comparing them with the original Parker sandstone, Leopard sandstone, carbonate shale, Berea sandstone, and sandy medium images. We evaluated the reconstruction by measuring pore and throat properties, two-point probability function, and structural similarity index. Results show that ACRNN can reconstruct reservoir rock or porous media of different scales with approximately the same geometrical, statistical, and topological parameters of the reservoir rock images. This deep learning method is computationally efficient, fast, and reliable for synthetic image realizations. The model was trained and validated on real images, and the reconstructed images showed excellent concordance with the real images having almost the same pore and grain structures. The deep learning-based digital rock reconstruction of reservoir rock or porous media images can aid in rapid image generation to better understand reservoir rock or subsurface formation.

Fatty Acid β-Oxidation May Be Associated with the Erythropoietin Resistance Index in Stable Patients Undergoing Haemodialysis.

Lipid metabolism and adiponectin modulate erythropoiesis in vitro and in general population studies and may also affect responsiveness to erythropoietin in patients undergoing haemodialysis (HD). However, little is known about the impact of lipid-associated biomarkers on reticulocyte production and erythropoietin resistance index (ERI) in patients undergoing HD. Therefore, we aimed to investigate their impacts in 167 stable patients undergoing HD. Pre-dialysis blood samples were collected and analysed for reticulocyte counts and serum lipid profiles by routine analyses and serum carnitine profiles (C0-C18) by LC-MS/MS. ERI was calculated as erythropoietin dose/kg/week normalized for haemoglobin levels. The independent positive determinants of reticulocyte count were log [Triglyceride (TG)] and logC18:1. A large proportion of longer-chain acylcarnitines was positively correlated with reticulocyte counts, possibly resulting from the accumulation of acylcarnitines in mitochondria undergoing fateful exocytosis from reticulocytes. These results indicate a possible association between reticulocyte formation and reduced β-oxidation, which occurs during the peripheral phase of erythroblast enucleation. Total cholesterol (TC) and log [C2/(C16 + C18:1)] as a putative marker of β-oxidation efficiency were negative independent determinants of ERI. Moreover, acyl chain length had a significantly positive impact on the correlation coefficients of individual acylcarnitines with ERI, suggesting that enhanced β-oxidation may be associated with reduced ERI. Finally, adiponectin had no independent association with reticulocyte counts or ERI despite its negative association with HDL-C levels. Enhanced fatty acid β-oxidation and higher TC levels may be associated with lower ERI, whereas higher TG levels and longer acylcarnitines may be related to the latest production of reticulocytes in stable patients undergoing HD.

Localization and discrimination of GG mismatch in duplex DNA by synthetic ligand-enhanced protein nanopore analysis.

Mismatched base pairs in DNA are the basis of single-nucleotide polymorphism, one of the major issues in genetic diseases. However, the changes of physical and chemical properties of DNA caused by single-site mismatches are often influenced by the sequence and the structural flexibility of the whole duplex, resulting in a challenge of direct detection of the types and location of mismatches sensitively. In this work, we proposed a synthetic ligand-enhanced protein nanopore analysis of GG mismatch on DNA fragment, inspired by in silico investigation of the specific binding of naphthyridine dimer (ND) on GG mismatch. We demonstrated that both the importing and unzipping processes of the ligand-bound DNA duplex can be efficiently slowed down in α-hemolysin nanopore. This ligand-binding induced slow-down effect of DNA in nanopore is also sensitive to the relative location of the mismatch on DNA duplex. Especially, the GG mismatch close to the end of a DNA fragment, which is hard to be detected by either routine nanopore analysis or tedious nanopore sequencing, can be well differentiated by our ND-enhanced nanopore experiment. These findings provide a promising strategy to localize and discriminate base mismatches in duplex form directly at the single-molecule level.

Multi-Characteristic Opsin Therapy to Functionalize Retina, Attenuate Retinal Degeneration, and Restore Vision in Mouse Models of Retinitis Pigmentosa.

Retinal degeneration 1 and 10 (rd1 and rd10) mice are useful animal models of retinitis pigmentosa (RP) with rapidly and slowly progressive pathologies, respectively. Our study aims were to determine the effect of adeno-associated viral vector 2 (AAV2)-delivered multi-characteristic opsin (MCO-010; under the control of a metabotropic glutamate receptor-6 promoter enhancer) on the morphological and functional characteristics of vision in both rd1 and rd10 mice. Various retinal measures of MCO-010 transduction and electrophysiological, behavioral, and other routine blood analyses were performed in the rd1 and/or rd10 mice after intravitreal injection of 1µL of MCO-010 or AAV2 vehicle. Functional tests included electroretinogram, visually evoked potential, and behavior assay (optomotor and water maze). Retinal thickness, intraocular pressure, and plasma cytokine levels were also determined. Following intravitreal MCO-010 injection, approximately 80% of bipolar cells were transduced in the retina, and no alterations in retinal thickness were observed at 4months post-injection. However, retinal thickness significantly decreased in control mice. MCO-010 treatment increased head movements and induced faster navigation of mice to the platform in a water-maze test. The MCO-010 gene therapy helped preserve visually evoked electrical response in the retina and visual cortex. No ocular toxicity, immunotoxicity, or phototoxicity was observed in the MCO-010-treated mice, even under chronic intense light conditions. Intravitreal MCO-010 was well tolerated in rd1 and rd10 mice models of RP, and it appeared to attenuate retinal photoreceptor degeneration based on retinal structure and functional outcome measures. As reported here, optogenetic treatment of the inner retina attenuates further retinal degeneration in addition to photosensitizing higher order neurons, and this disease-modifying aspect should be evaluated in optogenetic clinical trials.

Gut Analysis Toolbox - automating quantitative analysis of enteric neurons.

The enteric nervous system (ENS) consists of an extensive network of neurons and glial cells embedded within the wall of the gastrointestinal (GI) tract. Alterations in neuronal distribution and function are strongly associated with GI dysfunction. Current methods for assessing neuronal distribution suffer from undersampling, partly due to challenges associated with imaging and analyzing large tissue areas, and operator bias due to manual analysis. We present the Gut Analysis Toolbox (GAT), an image analysis tool designed for characterization of enteric neurons and their neurochemical coding using two-dimensional images of GI wholemount preparations. GAT is developed in Fiji, has a user-friendly interface, and offers rapid and accurate segmentation via custom deep learning (DL)-based cell segmentation models developed using StarDist, as well as a ganglia segmentation model in deepImageJ. We apply proximal neighbor-based spatial analysis to reveal differences in cellular distribution across gut regions using a public dataset. In summary, GAT provides an easy-to-use toolbox to streamline routine image analysis tasks in ENS research. GAT enhances throughput, allowing rapid unbiased analysis of larger tissue areas, multiple neuronal markers and numerous samples.

Clinical Utility of Recently FDA Approved IntelliSep test (Sepsis biomarker) for early Diagnosis of Sepsis: Comparison with other Biomarkers and von Willebrand Factor/ADAMTS13 Ration

Abstract Background Sepsis, a relatively common presentation in emergency departments, is a life-threatening disease. Recently, IntelliSep test by Cytovale (San Francisco, CA) received FDA approval as sepsis biomarker. We investigated clinical utility of this test using 44 patients (19 disease group and 25 non-disease group). Materials and Methods IntelliSep assesses cellular host response via deformability cytometry of leukocyte biophysical properties and intended for use in conjunction with clinical assessments and laboratory findings for early detection of sepsis. Test results are available within 10 minutes. Results consists of IntelliSep Band 1 (low risk, score: 0.1-4.9) to Band 3 (high risk, score: 6.3-10; total scale: 0.1-10). We measured IntelliSep score in 44 patients using EDTA blood. Left over plasma was used for measuring both plasma von Willebrand factor (vWF) antigen and ADAMTS13 antigen by ELISA assays. Monocyte distribution width (MDW) were obtained during routine CBC analysis using Beckman hematology analyzer. Lactate and high sensitivity troponin I were measured using Beckman analyzer. Results The median IntelliSep score was two-fold higher in the disease group (mean score 7.1, SD: 1.5, median: 7.4) compared to non-disease group (mean score 4.0, SD: 1.4, Median: 3.7) indicating that this new test can identify patients with high risk of developing sepsis and also rule out low risk patients (p<0.01 by both Mann-Whitney U test two tailed and t-test two tailed). The median MDW was also 34.8 % higher in the disease group (mean:25.6, SD: 5.0, Median: 24.4) compared to non-disease group (mean: 18.9, SD: 2.4, Median: 18.1) and difference was also statistically significant (p < 0.01 by both Mann-Whitney U test and t-test, two tailed). However, correlation between IntelliSep scores and MDW was only modest (overall, r= 0.66). Both lactate and procalcitonin concentrations were significantly higher in the disease group compared to the non-disease group. Interestingly, high-sensitivity troponin I concentrations were also significantly higher in the disease group. In addition, we observed significantly higher plasma vWF antigen in the disease group compared to the non-disease group (mean: 67.4 ug/mL in disease group vs 34.1 ug/mL in the non-disease group. However, in some patients in the disease group plasma vWF values were 6.8-fold higher than normal. In addition, vWF/ADAMTS13 ratio was also significantly elevated in the disease group compared to non-disease group. Discussion New IntelliSep test is useful in early prediction of sepsis in patients admitted to emergency department and may be superior to MDW and lactate for early diagnosis. Significant increase in plasma vWF in these patients indicate that vWF may also be a surrogate marker for endotheliopathy in sepsis. Lack of correlation between plasma VWF and Intellisep results indicates that these two markers probably measure different pathways for sepsis (endothelial activation vs neutrophil activation).

Lipid and Corticosteroid Biomarkers Under the Influence of Bisphosphonates.

Detecting the use of bisphosphonates (BPs) in equine athletes is of interest to regulators and laboratories due to the threat to welfare issues for the potential to provide analgesic effects and manipulating bone structure. The detection of BPs in biological matrices is challenging due to erratic biological elimination and inconsistent analytical recoveries. Therefore, complementary approaches are needed to provide evidence of their misuse in racehorses. BPs have two sub-classes: nitrogenous and non-nitrogenous. This study investigated plasma elimination following administration of one example from each sub-class, together with changes in endogenous eicosanoid and corticosteroids. Zoledronic acid (ZA) and tiludronic acid (TA) were administered by IV infusion to 8 thoroughbred horses with an 11-month washout period between each administration. Sample preparation for quantification of BPs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) utilised a two-step solid phase extraction (SPE) consisting of polymeric reversed-phase followed by weak anion exchange prior to derivatisation using trimethyl orthoacetate. Endogenous biomarkers were analysed after protein precipitation and SPE with polymeric reversed-phase prior to liquid chromatography-high resolution mass spectrometry (LC-HRMS) using data independent acquisition. The LC-MS/MS analysis showed ZA was undetectable after 8 h post-administration while TA was detected up to the final collection point of 28 days post-administration. The LC-HRMS analysis utilised targeted (i.e., prior inclusion list of compounds) approaches to monitor level changes of eicosanoid and corticosteroid biomarkers. Putative biomarkers were identified and now subject to validation for translation into routine sample analysis for improved retrospectivity to detecting BP misuse in equine plasma.

First fluorescence method for monitoring the doping stimulant drug solriamfetol in plasma and urine: A pharmacokinetic study in volunteers’ urine with greenness, whiteness, and blueness assessments

A green, sustainable, simple, and highly sensitive spectrofluorimetric method was developed to determine solriamfetol hydrochloride for the first time in biological fluids and dosage form. To the best of our knowledge, no Fluorimetric methods have been reported for solriamfetol determination in plasma and urine. The method is based on measuring the intrinsic fluorescence of solriamfetol hydrochloride at 522 nm after excitation at 260 nm in ammonium acetate buffer solution at pH 4. All factors influencing the response have been carefully investigated. The method was linear over a concentration range of 10 to 1800 ng/mL (r = 0.9998). The limit of detection (LOD) and limit of quantification (LOQ) values were found to be 3.2 and 9.8 ng/mL, respectively, indicating the excellent sensitivity of the proposed method. The pharmaceutical dosage form, spiked human plasma, and urine samples were successfully analyzed using the described method with average recoveries of 99.79 ± 0.739, 99.27 ± 1.885, and 99.57 ± 1.701, respectively. Plasma samples were first subjected to protein precipitation using 10 % perchloric acid, while urine samples were analyzed directly without any pretreatment. Furthermore, the suggested method was applied effectively on real human volunteers’ urine to measure renal clearance, the cumulative amount of solriamfetol excreted unchanged in the urine, and the percent of the dose recovered. The eco-friendly nature and sustainability of this work were evaluated using trending greenness, whiteness, and blueness assessment metrics. The greenness assessment tools, namely the Analytical GREEnness metric tool (AGREE), Analytical Eco-scale, and complementary green analytical procedure index (complex GAPI) were selected to prove the eco-friendly nature of the method. Additionally, whiteness and blueness assessments were conducted using the Red-Green-Blue model (RGB model), and high Blue Applicability Grade Index (BAGI), respectively. The proposed method is a promising analytical tool for routine analysis in quality control labs, therapeutic drug monitoring, and pharmacokinetics studies.

The predictive role of the TAPSE/sPAP ratio for cardiovascular events and mortality in systemic sclerosis with pulmonary hypertension.

Reduced TAPSE/sPAP ratio has recently emerged as a predictive parameter risk factor for PH, however its role in SSc has been poorly investigated. The aim of the study was to investigate the prognostic value of the TAPSE/sPAP ratio for the prediction of mortality and cardiovascular events in patients with SSc complicated by PH. A comparison between SSc patients with PAH (SSc-PAH) and those with PH and significant ILD (SSc-PH) was also carried out. A retrospective single-center study in which all patients having SSc-complicated by PH-referring to the Scleroderma-Unit of the AOU Policlinico of Modena, from October 2013 to October 2023 were evaluated. All SSc patients underwent recurrent clinical examination, routine blood chemistry analysis, functional, instrumental evaluation. 61 SSc patients (F/M 52/9) were enrolled. During the follow-up, 60.1% of patients experienced at least one cardiovascular event and 62% died. The main causes of death were PH (39.4%) and other heart-related events (39.4%). The TAPSE/sPAP ratio was significantly lower in deceased patients compared to survivors (mm/mmHg 0.3 ± 0.12SD vs. 0.48 ± 0.17SD, p < 0.001). Compared to the SSc-PAH subgroup, the SSc-PH patients had lower survival rates (55.3 ± 31.2 SD months vs. 25 ± 19 SD, p = 0,05). At the multivariate analysis, TAPSE/sPAP ratio <0.32 mm/mmHg, male gender, and the presence of significant ILD were identified as independent predictors of mortality and cardiovascular events. Our work confirmed the predictive role of the TAPSE/sPAP ratio for mortality and cardiovascular events in patients with SSc complicated by PH.

Enhancing low-level tritium detection in environmental waters: assessing the Hidex ULLA liquid scintillation counter

Tritium (³H) is a vital tracer in isotope hydrology and environmental studies, necessitating accurate and precise low-level detection methods. This study evaluated the performance of the new Hidex ULLA Liquid Scintillation Counter for ultra-low-level tritium assays in environmental water samples. The optimization of instrumental parameters enabled accurate and reproducible 3H measurements. Test samples (10–132 TU) were analyzed in triplicate, including 3H-free water and NIST traceable standards. The results demonstrated excellent accuracy and reproducibility, as evidenced by Zeta scores and linearity across these tritium concentrations. The system's stability over extended measurement time periods ensured reliable performance for routine ultra-low level 3H analysis. The optimization of key parameters such as coincidence time and region of interest (ROI) achieved the highest figure of merit (FoM) by using a 25 ns coincidence time and the DigPb open mode for optimal spectral fittings. Detection limits (DL) based on ISO11929 recommendations were significantly improved but highlighted the need for sample enrichment for most low-level environmental applications, with LSC counting times preferably exceeding 1000 min. Overall, the Hidex ULLA Liquid Scintillation Counter proved to be a robust and reliable instrument suitable for ultra-low-level tritium detection in environmental water samples.