Macromolecule-Nanoparticle-Based Hybrid Materials for Biosensor Applications.

Biosensors function as sophisticated devices, converting biochemical reactions into electrical signals. Contemporary emphasis on developing biosensor devices with refined sensitivity and selectivity is critical due to their extensive functional capabilities. However, a significant challenge lies in the binding affinity of biosensors to biomolecules, requiring adept conversion and amplification of interactions into various signal modalities like electrical, optical, gravimetric, and electrochemical outputs. Overcoming challenges associated with sensitivity, detection limits, response time, reproducibility, and stability is essential for efficient biosensor creation. The central aspect of the fabrication of any biosensor is focused towards forming an effective interface between the analyte electrode which significantly influences the overall biosensor quality. Polymers and macromolecular systems are favored for their distinct properties and versatile applications. Enhancing the properties and conductivity of these systems can be achieved through incorporating nanoparticles or carbonaceous moieties. Hybrid composite materials, possessing a unique combination of attributes like advanced sensitivity, selectivity, thermal stability, mechanical flexibility, biocompatibility, and tunable electrical properties, emerge as promising candidates for biosensor applications. In addition, this approach enhances the electrochemical response, signal amplification, and stability of fabricated biosensors, contributing to their effectiveness. This review predominantly explores recent advancements in utilizing macrocyclic and macromolecular conjugated systems, such as phthalocyanines, porphyrins, polymers, etc. and their hybrids, with a specific focus on signal amplification in biosensors. It comprehensively covers synthetic strategies, properties, working mechanisms, and the potential of these systems for detecting biomolecules like glucose, hydrogen peroxide, uric acid, ascorbic acid, dopamine, cholesterol, amino acids, and cancer cells. Furthermore, this review delves into the progress made, elucidating the mechanisms responsible for signal amplification. The Conclusion addresses the challenges and future directions of macromolecule-based hybrids in biosensor applications, providing a concise overview of this evolving field. The narrative emphasizes the importance of biosensor technology advancement, illustrating the role of smart design and material enhancement in improving performance across various domains.

Development and Validation of Novel HPLC Methods for Quantitative Determination of Vitamin D3 in Tablet Dosage Form.

In the present work, an efficient isocratic HPLC method was developed for the precise and accurate estimation of vitamin D3 in tablet form. The chromatographic conditions comprised an L3 silica column (5 µm in particle size, 4.6 mm × 250 mm) with a mobile phase n-hexane/ethyl acetate (85:15 v/v) with a flow rate of 2.0 mL/min and a detection wavelength of 292 nm. The new methodology was validated for accuracy, precision, specificity, robustness, and quantification limits according to an official monograph of USP/BP and ICH guidelines. The peak areas of the six replicates of the homogeneous sample were recorded. The mean value obtained was 67,301, and the relative standard deviation (RSD) was 0.1741. The linearity and range were in the acceptable bounds, i.e., 0.999, which was calculated using regression line analysis. The results show that the method is truly acceptable as the RSD, as the flow rate was 0.81%, while for the mobile phase composition, it was 0.72%, which lies in the acceptable range. The limit of detection (LOD) and the limit of quantification (LOQ) values were 0.0539 µg/mL and 0.1633 µg/mL, respectively. The % RSD of the intra and inter-day precision of the method was deemed acceptable according to the international commission for harmonization guidelines. The developed method has potential to be used for the detection and quantification of vitamin D3 during routine analysis for tablets in dosage form.

Using QuEChERS and HPLC Method to Monitor the Background Concentration of Polycyclic Aromatic Hydrocarbons in Commercial Black Tea Leaves and Infusions in Taiwan.

Tea is an integral part of Taiwanese culture and is a popular drink as it contains many beneficial compounds. However, during the processing of tea, polycyclic aromatic hydrocarbons (PAHs) may form. This study investigated the concentrations of PAH4 in different black tea leaves and tea infusions based on the origin of the tea. The samples were extracted using QuEChERS, while the content of PAH4 was analyzed by high performance liquid chromatography coupled to a fluorescence detector (HPLC-FLD). The content of PAH4 in the tea leaves ranged from 2.88 µg/kg to 218.2 µg/kg (dry weight), with the highest concentration being found in teas from Vietnam. The concentration of BaP ranged from ND to 47.92 µg/kg. The release of PAH4 from tea leaves to tea infusions was significantly low, with the highest transfer being 25.8%. In this study, all PAH4 compounds in commercial black tea leaves can be detected by QuEChERS extraction with a simple HPLC method.

Summary recommendations on "Analytical methods for substances in the Watch List under the Water Framework Directive".

The Watch List (WL) is a monitoring program under the European Water Framework Directive (WFD) to obtain high-quality Union-wide monitoring data on potential water pollutants for which scarce monitoring data or data of insufficient quality are available. The main purpose of the WL data collection is to determine if the substances pose a risk to the aquatic environment at EU level and subsequently to decide whether a threshold, the Environmental Quality Standards (EQS) should be set for them and, potentially to be listed as priority substance in the WFD. The first WL was established in 2015 and contained 10 individual or groups of substances while the 4th WL was launched in 2022. The results of monitoring the substances of the first WL showed that some countries had difficulties to reach an analytical Limit of Quantification (LOQ) below or equal to the Predicted No-Effect Concentrations (PNEC) or EQS. The Joint Research Centre (JRC) of the European Commission (EC) organised a series of workshops to support the EU Member States (MS) and their activities under the WFD. Sharing the knowledge among the Member States on the analytical methods is important to deliver good data quality. The outcome and the discussion engaged with the experts are described in this paper, and in addition a literature review of the most important publications on the analysis of 17-alpha-ethinylestradiol (EE2), amoxicillin, ciprofloxacin, metaflumizone, fipronil, metformin, and guanylurea from the last years is presented.

Quantification of pendimethalin residue in green fodder and silage of winter cereals using gas chromatography-tandem mass spectrometry.

Use of agro-chemicals in forage crops is restricted due to the fear of direct toxicity to livestock and risk of pesticide residue accumulation in the food chain. Wheat and barley can be used as green fodder and silage, and herbicide residue estimation in green fodder and silage is important for ensuring the safety of dairy cattle. A field experiment was conducted for two years to study pendimethalin residues in the green fodder and silage of wheat and barley. In both cereal crops, pendimethalin (1.125 kg a.i./ha) was applied as pre-emergence along with an unsprayed control. Pendimethalin residues in fodder, silage, and soil were estimated using gas chromatography-tandem mass spectrometry (GC-MS/MS). At harvest, pendimethalin residues in fodder and silage of wheat and barley were below the limit of quantification (<0.01 mg/kg) during both crop seasons. Pendimethalin can be safely used for weed control in winter cereals grown for fodder and silage.

Quantification of Ochratoxin A in 90 spice and herb samples using the ELISA method.

This study addressed the challenge of accurately detecting mycotoxins in herbs and spices, which have gained popularity as alternative medicines but pose health risks due to potential contamination. We used a competitive direct ELISA kit (Art No. 8610), Veratox for Ochratoxin, to quantify Ochratoxin A in the herb and spice samples. The samples were first prepared using solid-liquid extraction with 70% methanol. The resulting filtrate was then subjected to ELISA analysis. The results of the analysis were then further analyzed using principal component analysis (PCA). In this study, PCA was used to classify the concentration levels of Ochratoxin A based on various factors, such as the packaging type, country of origin, shelf life, and sample weight. The limits of detection (LOD) and quantification (LOQ) values indicate the lowest amount of Ochratoxin A that can be detected and quantified, respectively, with high accuracy and precision. The range of the LOD and LOQ values (0.43-0.58 µg/kg and 1.45-1.95 µg/kg, respectively) suggests that the method used was capable of detecting and quantifying Ochratoxin A in the herb and spice samples at different concentrations with a high degree of accuracy and precision. These results suggest that while most of the samples (73.33%) were below the maximum residue limit (MRL) for Ochratoxin A, a significant number of samples (26.67%) had concentrations of Ochratoxin A that were higher than the MRL. This highlights the importance of monitoring Ochratoxin A in herb and spice samples and ensuring the products are safe for consumption.

Evaluating Proteomics Imputation Methods with Improved Criteria.

Quantitative measurements produced by tandem mass spectrometry proteomics experiments typically contain a large proportion of missing values. Missing values hinder reproducibility, reduce statistical power, and make it difficult to compare across samples or experiments. Although many methods exist for imputing missing values, in practice, the most commonly used methods are among the worst performing. Furthermore, previous benchmarking studies have focused on relatively simple measurements of error such as the mean-squared error between imputed and held-out values. Here we evaluate the performance of commonly used imputation methods using three practical, "downstream-centric" criteria. These criteria measure the ability to identify differentially expressed peptides, generate new quantitative peptides, and improve the peptide lower limit of quantification. Our evaluation comprises several experiment types and acquisition strategies, including data-dependent and data-independent acquisition. We find that imputation does not necessarily improve the ability to identify differentially expressed peptides but that it can identify new quantitative peptides and improve the peptide lower limit of quantification. We find that MissForest is generally the best performing method per our downstream-centric criteria. We also argue that existing imputation methods do not properly account for the variance of peptide quantifications and highlight the need for methods that do.

Evaluation of Statistical Treatment of Left-Censored Contamination Data: Example Involving Deoxynivalenol Occurrence in Pasta and Pasta Substitute Products.

The handling of data on food contamination frequently represents a challenge because these are often left-censored, being composed of both positive and non-detected values. The latter observations are not quantified and provide only the information that they are below a laboratory-specific threshold value. Besides deterministic approaches, which simplify the treatment through the substitution of non-detected values with fixed threshold or null values, a growing interest has been shown in the application of stochastic approaches to the treatment of unquantified values. In this study, a multiple imputation procedure was applied in order to analyze contamination data on deoxynivalenol, a mycotoxin that may be present in pasta and pasta substitute products. An application of the proposed technique to censored deoxynivalenol occurrence data is presented. The results were compared to those attained using deterministic techniques (substitution methods). In this context, the stochastic approach seemed to provide a more accurate, unbiased and realistic solution to the problem of left-censored occurrence data. The complete sample of values could then be used to estimate the exposure of the general population to deoxynivalenol based on consumption data.

Application of Static Headspace GC-MS Method for Selective 1,4-Dioxane Detection in Food Additives.

Efficient detection methods must be developed for 1,4-dioxane due to its suspected status as a human carcinogen, which is highly mobile in food and environmental resources. In this regard, this experiment has been conducted to develop reliable and selective detection and measurement methods by using static headspace (SH) isolation, followed by gas chromatography-mass spectrometry (GC-MS). A new method was developed for determining the spiked 1,4-dioxane contents in a polyethylene glycol 600 (PEG 600). The optimal condition for SH-GC-MS was discussed. The representative ions of 1,4-dioxane and 1,4-dioxane-d8 in the SIM mode of MS are 88 and 96, respectively, and the peaks of the SIM mode were separated and confirmed. The linear range for the method covers 0.25 to 100 mg/L with a coefficient of determination (R2) ≥ 0.999. The method applicability was demonstrated by spike recovery across a variety of food additives (i.e., chlorine bitartrate, choline chloride, polysorbate 20 and 60, and PEG 1000). All spike recovery from the tested samples was in the range of 89.50-102.68% with a precision of 0.44-11.22%. These findings suggest a new analytical method for food safety inspection, and could be applicable for ensuring the safety of foods and environmental and public health on a broad scale.

Inferring the distribution of norovirus in individual oysters below the limit of quantification by pooled sampling.

Norovirus (NoV) in oysters is a food safety risk of much concern. In order to assess the risk of the exposure, the distribution of the number of NoV copies contained in each oyster should be acquired first for comprehensively quantifying the associated risks. However, the part of the distribution below the limit of quantification cannot be obtained directly by laboratory detecting methods, which hampers accurate assessment. To tackle this challenging problem, a systematic method (Distribution Inference Method by Pooled Sampling) is proposed to infer the unobservable part of distribution based upon all measurements of the pooled samples with n = 2. Using convolutional integrals and real-coded genetic algorithm for inferring, this method has neither requirements for the type or properties of the original distribution, nor requirements for historical data, even nor requirements for the relationship between observable and unobservable parts of the distribution. A series of experiments were conducted on simulated datasets of a variety of types, including normal distribution, uniform distribution, gamma distribution, lognormal distribution, zero-inflated Poisson distribution, their combinations, and even their splicing, covering common distribution types in oyster NoV scenario and more general scenarios. The results show that almost all inferred simulation data and their original counterparts passed Kolmogorov-Smirnov tests, which implies that they are essential of the same distribution. Based on this method, a ready-to-use web system was developed for researchers to infer their original distribution with pooled-sampling measurements from the detection of NoV or even other substances.

Consideration of unmeasured micropollutants released from WWTP for potential impact estimations.

During wastewater treatment, micropollutants are only partly eliminated and may present a risk for human health and aquatic ecosystems. The potential impacts these substances may have are currently underestimated due to the lack in available concentrations that lie below the limit of quantification (LOQ) for an important set of micropollutants. Here, the potential impacts due to 261 organic micropollutants on human health and aquatic environments were investigated at the scale of France. Even with concentrations below the LOQ, certain micropollutants were found to have a significant potential impact. For unmeasured concentrations, a global concentration distribution built from several datasets with different LOQ was used. By disregarding the unmeasured micropollutants, the potential impacts have been underestimated by >300 % on both human health and aquatic environments. Certain substances, such as hydrazine, endrin, or 2,3,7,8-TetraCDD, could lead to very strong potential impacts, even with unmeasured concentration levels. Moreover, the usual convention of LOQ/2 to replace unmeasured concentrations also appeared to overestimate the potential impact. The present work can be adapted to any other compartment or geographical context.

A systematic review and meta-analysis of validated analytical techniques for the determination of total selenium in foods and beverages.

The intricate balance between the beneficial and harmful effects of selenium (Se) intake means that its quantification in food needs to be done correctly. Therefore, in this review, we systematized 105 articles to identify the most studied methodologies, analytical techniques, and food matrices. Among the analytical techniques employed, inductively coupled plasma mass spectrometry (ICP-MS) (n = 29) emerged as the most commonly used method. The most prevalent hydrolysis methodology to digest Se in food matrices involved the use of nitric acid combined with ultrasound, which improved both the yield and digestion time. Optimal recovery values were achieved when total Se quantification accounted for the sum of Se(IV) and Se(VI) (94.4-99.4%) and for SeCys (88-96.5%). These findings are relevant for advancing methodological approaches, and their results emphasize the importance of developing alternative, faster, and lower-cost protocols for Se quantification in foods and beverages.

Quantitative PCR versus metagenomics for monitoring antibiotic resistance genes: balancing high sensitivity and broad coverage.

The widespread occurrence of clinically relevant antibiotic resistance within humans, animals, and environment motivates the development of sensitive and accurate detection and quantification methods. Metagenomics and quantitative PCR (qPCR) are amongst the most used approaches. In this study, we aimed to evaluate and compare the performance of these methods to screen antibiotic resistance genes in animal faecal, wastewater, and water samples. Water and wastewater samples were from hospital effluent, different treatment stages of two treatment plants, and of the receiving river at the discharge point. The animal samples were from pig and chicken faeces. Antibiotic resistance gene coverage, sensitivity, and usefulness of the quantitative information were analyzed and discussed. While both methods were able to distinguish the resistome profiles and detect gradient stepwise mixtures of pig and chicken faeces, qPCR presented higher sensitivity for the detection of a few antibiotic resistance genes in water/wastewater. In addition, the comparison of predicted and observed antibiotic resistance gene quantifications unveiled the higher accuracy of qPCR. Metagenomics analyses, while less sensitive, provided a markedly higher coverage of antibiotic resistance genes compared to qPCR. The complementarity of both methods and the importance of selecting the best method according to the study purpose are discussed.

Statement on the risk assessment of maximum residue levels (MRLs) for oxamyl in view of consumer protection.

In accordance with Article 43 of Regulation (EC) No 396/2005, the European Commission requested EFSA to perform a risk assessment of the existing maximum residues levels (MRLs) for oxamyl considering the new toxicological reference values. Additionally, if needed to ensure adequate consumer protection, lower limits of quantification (LOQs) than those currently established in the legislation should be proposed. EFSA performed various consumer exposure calculation scenarios, considering the risk assessment values as available for the existing uses of oxamyl and the lowering of LOQs for several plant and animal commodities as suggested by the European Union Reference Laboratories for Pesticide Residues (EURLs). Based on the results of the consumer exposure assessment calculated considering the risk assessment values for crops with authorised oxamyl uses and the existing EU MRLs at the LOQ for remaining commodities (scenario 1), chronic consumer intake concerns were identified for 34 diets. Acute exposure concerns were identified for a wide range of crops, including crops with currently authorised oxamyl uses: bananas, potatoes, melons, cucumbers, carrots, watermelons, tomatoes, courgettes, parsnips, salsifies and aubergines/eggplants. Under exposure calculation scenario 3, which considered lowering of all MRLs to the lowest analytically achievable limits of quantification, EFSA concludes that chronic consumer exposure concerns can still not be excluded. Similarly, acute consumer exposure concerns were identified for 16 commodities, including crops with known authorised uses: potatoes, melons, watermelons and tomatoes, even though for these crops a lower LOQ as proposed by the EURLs were considered. Further refinements of the calculated exposure at the current stage were not possible by EFSA, but EFSA identified a list of commodities for which a lower LOQ than routinely achievable is expected to significantly reduce the consumer exposure and for which a risk management decision is required.

Chemical and in vitro toxicological comparison of emissions from a heated tobacco product and the 1R6F reference cigarette.

It has previously been found that, compared with cigarette smoke, the aerosols generated by heated tobacco products contain fewer and lower harmful and potentially harmful constituents (HPHCs) and elicit lower biological activity in in vitro models and lower smoking-related exposure biomarker levels in clinical studies. It is important to accumulate such scientific evidences for heated tobacco products with a novel heating system, because different heating system may affect the quantitative aspect of the amount of HPHCs and the qualitative aspect of the biological activity of the aerosol generated. Here, the chemical properties of, and toxicological responses to aerosols emitted by DT3.0a, a new heated tobacco product with a novel heating system, and cigarette smoke (CS) were compared, using chemical analyses, in vitro battery (standardized genotoxicity and cytotoxicity) assays, and mechanistic (ToxTracker and two-dimensional cell culture) assays. Regular- and menthol-flavored DT3.0a and standard 1R6F reference cigarettes were tested. Selected HPHC yields were lower in DT3.0a aerosol than 1R6F CS. The genotoxicity-related assays indicated that DT3.0a aerosol was not genotoxic, regardless of metabolic activation. The other biological assays indicated that less cytotoxicity induction and oxidative stress response were elicited by DT3.0a aerosol compared with 1R6F CS. Similar results were found for both regular and menthol DT3.0a. Like previous reports for heated tobacco products with other heating systems, the results of this study indicated that DT3.0a aerosols have chemical and biological properties less likely to be harmful than 1R6F CS.

An integrated human health risk assessment framework for alkylphenols due to drinking water and crops' food consumption.

The increasing overexploitation and pollution of freshater resources are potential threats for public health, causing cross-contamination among the interconnected environmental compartments (freshwater, soil, crops). In particular, contaminants of emerging concern (CECs) originating from anthropic activities are not completely removed by wastewater treatments plants. This leads to their presence in drinking water (DW) sources, soil and crops intended for human consumption due to discharges of treated wastewater in surface waters and direct wastewater reuse practices. Currently, health risk assessments are limited to single exposure sources without considering the multiple exposure routes to which humans are subjected. For instance, among CECs, bisphenol A (BPA) and nonylphenol (NP), respectively, adversely affect immune and renal systems and have been frequently detected in DW and food, their major exposure sources for humans. Here, an integrated procedure is proposed to quantitatively assess health risk from CECs due to multiple exposure from the consumption of both DW and food, considering the relevant inter-connected environmental compartments. This procedure was applied to BPA and NP to calculate their probabilistic Benchmark Quotient (BQ), showing its potential in quantitatively apportioning the risk between contaminants and exposure sources, and its use as a decision support tool for prioritizing mitigation measures. Our results indicate that, even though the human health risk due to NP is not negligible, the estimated risk due to BPA is significantly higher, and the consumption of food from edible crops determines a higher risk compared to tap water. Hence, BPA is undoubtedly a contaminant to be prioritized, especially through mitigation actions aimed at its prevention and removal from food.

Pre-analytical sample handling standardization for reliable measurement of metabolites and lipids in LC-MS-based clinical research.

The emerging disciplines of lipidomics and metabolomics show great potential for the discovery of diagnostic biomarkers, but appropriate pre-analytical sample-handling procedures are critical because several analytes are prone to ex vivo distortions during sample collection. To test how the intermediate storage temperature and storage period of plasma samples from K3EDTA whole-blood collection tubes affect analyte concentrations, we assessed samples from non-fasting healthy volunteers (n = 9) for a broad spectrum of metabolites, including lipids and lipid mediators, using a well-established LC-MS-based platform. We used a fold change-based approach as a relative measure of analyte stability to evaluate 489 analytes, employing a combination of targeted LC-MS/MS and LC-HRMS screening. The concentrations of many analytes were found to be reliable, often justifying less strict sample handling; however, certain analytes were unstable, supporting the need for meticulous processing. We make four data-driven recommendations for sample-handling protocols with varying degrees of stringency, based on the maximum number of analytes and the feasibility of routine clinical implementation. These protocols also enable the simple evaluation of biomarker candidates based on their analyte-specific vulnerability to ex vivo distortions. In summary, pre-analytical sample handling has a major effect on the suitability of certain metabolites as biomarkers, including several lipids and lipid mediators. Our sample-handling recommendations will increase the reliability and quality of samples when such metabolites are necessary for routine clinical diagnosis.

A human monoclonal antibody against HBsAg for the prevention and treatment of chronic HBV and HDV infection.

Background & Aims: Elimination of chronic HBV/HDV infection remains a major global health challenge. Targeting excessive hepatitis B surface antigen (HBsAg) release may provide an interesting window of opportunity to break immune tolerance and to achieve a functional cure using additional antivirals. Methods: We evaluated a HBsAg-specific human monoclonal antibody, as part of either a prophylactic or therapeutic strategy, against HBV/HDV infection in cell culture models and in human-liver chimeric mice. To assess prophylactic efficacy, mice were passively immunized prior to infection with HBV or HBV/HDV (coinfection and superinfection setting). Therapeutic efficacy was assessed in HBV and HBV/HDV-coinfected mice receiving 4 weeks of treatment. Viral parameters (HBV DNA, HDV RNA and HBsAg) were assessed in mouse plasma. Results: The antibody could effectively prevent HBV/HDV infection in a dose-dependent manner with IC50 values of ∼3.5 ng/ml. Passive immunization showed complete protection of mice from both HBV and HBV/HDV coinfection. Moreover, HDV superinfection was either completely prevented or at least attenuated in HBV-infected mice. Finally, antibody treatment in mice with established HBV/HDV infection resulted in a significant decline in viremia and a concomitant drop in on-treatment HBsAg, with a moderate viral rebound following treatment cessation. Conclusion: We present data on a valuable antibody candidate that could complement other antivirals in strategies aimed at achieving functional cure of chronic HBV and HDV infection. Impact and implications: Patients chronically infected with HBV may eventually develop liver cancer and are at great risk of being superinfected with HDV, which worsens and accelerates disease progression. Unfortunately, current treatments can rarely eliminate both viruses from chronically infected patients. In this study, we present data on a novel antibody that is able to prevent chronic HBV/HDV infection in a mouse model with a humanized liver. Moreover, antibody treatment of HBV/HDV-infected mice strongly diminishes viral loads during therapy. This antibody is a valuable candidate for further clinical development.

Investigation of the photoactivation effect of TiO2 onto carbon-clay paste electrode by cyclic voltammetry analysis.

In this work, a cyclic voltammetry analysis for the detection of Ascorbic Acid (AA) based on a carbon-clay paste electrode modified with titanium dioxide (CPEA/TiO2) is presented. The electrochemical sensor was prepared using clay and carbon graphite, mixed with TiO2 to investigate the electrode behavior towards the detection of AA. Comprehensive characterization approaches including X-ray diffraction (XRD), Selected area electron diffraction (SAED), Transmission electron microscopy (TEM), Fourier transform infra-red spectroscopy (FTIR) were carried out on different samples. The results indicated that, the electrode has been effectively modified, while the electrochemical parameters of AA on CPEA/TiO2/UV such as the charge transfer coefficient (α a ), number of electrons (n) transferred and standard potential were calculated. CPEA/TiO2/UV exhibit better photoactivity and also higher electronic conductivity under light radiation (100 W). The linear range for AA was determined between 0.150µM and 0.850 µM with the straight-line equation equivalent to I p a ( µ A ) = 2.244 [ A A ] + 1.234 (n = 8, R2 = 0.993). The limit of detection was 0.732 µM (3σ) and limit of quantification was 2.440 µM. For the analytical applications, pharmaceutical tablets such as Chloroquine phosphate, Azithromycin and Hydroxychloroquine sulfate were performed. In addition, interference study in the analytical application was performed, and it was found that the electroanalytical method used can be well adopted for simultaneous electrochemical detection of AA and Azithromycin.

Analytical performance of Abbott C-16000 analyser haemolysis index and its potential use in measuring plasma cell-free haemoglobin.

Purpose: Measurement of the haemolysis index (HI) is usually performed in clinical chemistry laboratories in order to inform about whether biological analyses are influenced by in vivo or in vitro haemolysis of the specimen. Our aim was to evaluate the analytical performance of Abbott C-16000 analyser HI measurement in order to determine whether this could be used to reliably measure cell-free haemoglobin (fHB) in plasma samples. Methods: The repeatability, reproducibility, lower limit of detection (LLOD) and lower limit of quantification (LLOQ) of C-16000 HI measurement were determined as well as the potential interference of bilirubin, triglycerides and myoglobin. C-16000 HI values of biological samples with various ranges of fHB were compared to those measured using the established reference method, second-derivate spectroscopy. Results: Results: C-16000 HI determination showed excellent linear correlation with the reference method (y = 1.0043x ­ 1.248, R² = 0.998), a broad analytical measurement range (400-20,000 mg/L; y = 0.9904x + 72.972, R² = 0.999), clinically relevant LLOD (56 mg/L) and LLOQ (84 mg/L), good repeatability (coefficient of variation (CV) = 1-15%) and good reproducibility (CV = 5-7%). No interference was observed with myoglobin at concentrations as high as 35,447 mg/L, unconjugated and conjugated bilirubin (at concentrations up to 500 mg/L and 375 mg/L, respectively) or triglycerides up to 6.8 mmol/L. However, a significant underestimation of fHB concentrations was observed at higher triglyceride levels. Conclusion: This study demonstrates that Abbott C-16000 analyser HI is reliable and accurately measures plasma fHB concentrations under pathophysiological conditions except when there are high blood concentrations of triglycerides.