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Even though dilute (unentangled) polymer solutions cannot act as gel-like sieving media, it has been shown that they can be used to separate DNA molecules in capillary electrophoresis. The separation then comes from sporadic, independent DNA–polymer collisions. We study polymer–polymer collisions in nanochannels (i.e., channels that are smaller than the normal size of the polymers), a situation where a polyelectrolyte is forced to migrate "through" isolated uncharged molecules during electrophoresis. We use Langevin dynamics simulations to explore the nature of these collisions and their effect on the net motion of the two polymer chains. We identify several types of collisions, including some that are unique to nanochannels. When the uncharged polymer is much larger than the polyelectrolyte, the system is reminiscent of gel electrophoresis, and we propose a modified empirical reptation model to explain the data, with an orientation factor that depends on the tube diameter. We also observe that the duration of a collision is a non-monotonic function of the polymer size ratio when the two chains are of comparable size, a surprising resonance-like phenomenon, which, combined with the asymmetric nature of molecular conformations during collision, suggests possible ratchet-like mechanisms that could be used to sort polyelectrolytes in nanodevices. [ABSTRACT FROM AUTHOR]

A capillary zone electrophoresis (CZE) system was coupled to an Orbitrap mass spectrometer operating in a data-independent acquisition (DIA) mode for in-depth proteomics analysis. The performance of this CZE-DIA-MS system was systemically evaluated and optimized under different operating conditions. The performance of the fully optimized CZE-DIA-MS system was subsequently compared to the one by using the same CZE-MS system operating in a data-dependent acquisition (DDA) mode. The experimental results show that the numbers of identified peptides and proteins acquired in the DIA mode are much higher than the ones acquired in the DDA mode, especially with the small sample loading amount. Specifically, the numbers of identified peptides and proteins acquired in the DIA mode are 1.8-fold and 2-fold higher than the ones acquired in the DDA mode by using 12.5 ng Hela digests. The proteins identified in the DIA mode also cover almost all the proteins identified in the DDA mode. In addition, a potential cancer biomarker protein, carbohydrate antigen 125, undetected in the DDA mode, can be easily identified in the DIA mode even with 12.5 ng Hela digests. The performance of the CZE-DIA-MS system for in-depth proteomics analysis with a limited sample amount has been fully demonstrated for the first time through this study. [ABSTRACT FROM AUTHOR]

Understanding the genetic diversity of crops is of fundamental importance for the efficient use and improvement of germplasm resources. Different molecular genotyping systems have been implemented for population structure and phylogenetic relationships analyses, among which, microsatellites (SSRs) and single nucleotide polymorphisms (SNPs) markers have been the most widely used. This study reports the efficacy of SNPs detected via double-digest restriction-site-associated DNA sequencing (ddRADseq) and SSRs analyzed via capillary electrophoresis (CE) and high-resolution melting (HRM) in tomato. In total, 21,020 high-quality SNPs, 20 CE-SSRs, and 17 HRM-SSR markers were assayed in a panel of 72 accessions that included a diversified set of landraces, long-shelf-life cultivars and heirlooms with different origins and fruit typology. The results showed how the population structure analysis was consistent using the three genotyping methods, although SNPs were more efficient in distinguishing cultivar types and in measuring the degree of accessions' similarity. Compared to CE-SSR, the analysis of microsatellites via HRM yielded a slightly higher number of alleles (98 vs. 96). HRM-SSR demonstrated a distinction between European and non-European germplasm, better resolving the collection's diversity and being more consistent with SNP data. Phylogenetic trees drawn with independent marker data, detected specific groups of accessions showing robust clusters, highlighting how heirlooms were less heterogeneous than landraces. In addition, the fixation index (FST) revealed a high genetic differentiation between heirlooms and long-shelf-life cultivars, with SNP and SSR-HRM data emphasizing the distinction between cherry and plum types and CE-SSR data between cherry and oxheart types. In all instances, a greater molecular variance was found within the different considered biological statuses, provenances, and typologies rather than among them. This work presents the first attempt to compare the three tomato genotyping techniques in tomato. Findings highlighted how the markers used are complementary for genetic diversity analysis, with SNPs providing better insight and HRM-SSR as a viable alternative to capillary electrophoresis to dissect the genetic structure. [ABSTRACT FROM AUTHOR]

AbstractMeasuring plasma albumin is a common and important laboratory test. We compared the results obtained with the bromocresol green (BCG) colorimetric, immunoturbidimetric (IT), and capillary electrophoresis (CE) methods and evaluated the clinical reliability of the colorimetric test. Samples from 320 patients including 227 patients with hypoalbuminemia (albumin levels <35 g/L) were analyzed. Results were compared between different patient groups. The BCG method indicated significantly higher plasma albumin levels than the CE and IT methods, especially in patients with elevated C-reactive protein, alpha-1 globulin (a1G), and alpha-2 globulin (a2G) values. A significant proportion of patients with mild hypoalbuminemia tested using the BCG method (albBCG) and were classified as severely hypoalbuminemic (albumin <20 g/L) when switching to the CE or IT method (albCE and albIT). These patients had elevated a1G and/or a2G levels. This change of result implied an additional indication for albumin replacement therapy. The BCG method significantly overestimates albumin levels in patients with inflammation and hypoalbuminemia, which may lead to inappropriate therapeutic decisions. [ABSTRACT FROM AUTHOR]

Neonicotinoids (NNIs) are neuro-active and systemic insecticides widely used to protect crops from pest attack. During the last decades, there has been an increase concern about their uses and toxic effects, especially to beneficial and non-target insects such as pollinators. To assess potential health hazards and the environmental impacts derived from NNIs uses, a great variety of analytical procedures for the determination of their residues and their metabolites at trace level in environmental, biological and food samples have been reported. Due to the complexity of the samples, efficient sample pretreatment methods have been developed, which include mostly clean-up and preconcentration steps. On the other hand, among the analytical techniques used for their determination, high-performance liquid chromatography (HPLC) coupled to ultraviolet (UV) or mass spectrometry (MS) detection is the most widely used, although capillary electrophoresis (CE) has also been employed in the last years, considering some improvements in sensitivity when coupling with new MS detectors. In this review, we present a critical overview of analytical methods based on HPLC and CE reported in the last decade, discussing relevant and innovative sample treatments for the analysis of environmental, food and biological samples. [ABSTRACT FROM AUTHOR]

Erectile dysfunction (ED) is one of the most common chronic diseases affecting men and its incidence increases with aging. Due to its substantial influence on the quality of life, phosphodiesterase type-5 (PDE5) inhibitors have been implemented to treat ED by increasing the penile blood flow that results in improving erection. PDE5 inhibitors is a class of drugs that affects many pharmacological sectors, and it is essential to review the different analytical methods described for their determination. Few reviews were published concerning this group of drugs. For this reason, this review article gathers the different analytical methods used to determine PDE5 inhibitors in pharmaceutical and biological samples over the past 20 years. Different analytical techniques were used to analyze these compounds in different matrices such as separation methods (capillary electrophoresis, LC-MS, UPLC-MS/MS, and GC-MS), spectroscopic methods (UV-visible methods, FT-IR spectroscopy and spectrofluorometry) and electrochemical methods (polarography, voltammetry and potentiometry). This review focuses on the different electrochemical methods and their use in analytical determination of PDE5 inhibitors in pharmaceutical dosage forms and biological samples. Moreover, it discusses the different modified electrodes used for their electroanalytical determination and the behavior of the studied drugs at different modified electrodes. Additionally, this review discusses the pharmacokinetics of the studied compounds and their interactions with other co-administered drugs especially the metabolic interactions between the studied compounds and other co-administered drugs in different matrices. This literature survey would provide a beneficial guide for future analytical investigation of PDE5 inhibitors. [ABSTRACT FROM AUTHOR]

Dating silk mounting textiles is important for understanding their condition and susceptibility to degradation. Amino acid racemization is a technique currently used for this purpose. The research presented here aimed to clarify when this technique is reliable and when it is not. Silk fibers taken from a cocoon and from Japanese mounting textiles were studied using optical microscopy to document surface roughness. Chiral capillary electrophoresis mass spectrometry for amino acid racemization dating (AAR), liquid chromatography mass spectrometry for dye analysis, and FTIR for analysis of sericin and other materials adhering to the silk were also used. A change from rough fiber surfaces (greater than a hundred years old) to smooth fiber surfaces (less than a hundred years) was found. This correlated with the time of the introduction of Western degumming methods in Japan. Synthetic dyes were not found on the earlier silk fibers with rough surfaces. AAR dating measurements on rough surfaced fibers with remaining sericin correlated better with the estimated age of the silk fabrics than fibers with smooth surfaces, suggesting the technique should only be used for silk fibers with rough surfaces. A screening method for fibers for AAR analysis was outlined. [ABSTRACT FROM AUTHOR]

The reuse of the same chromatography resin to purify multiple products is of high economic and ecologic interest in the pharmaceutical industry. An effective cleaning program to ensure no product carryover from one product to the subsequent is key to enable a multi-product use strategy. The methods utilised to assess product carryover during resin lifetime studies for a given product are not specific enough to detect carryover to support multi-product use. Here the proposed use of liquid chromatography coupled to mass spectrometry (LC-MS) is the analytical method of choice due to its high sensitivity and selectivity. A proof-of-concept study using protein A resin and two distinct IgG product streams produced by CHO cell lines was conducted. The experimental design was developed to determine any carryover from previous products after a multiple product Protein A resin reuse process, using LC-MS and capillary electrophoresis (CE). The capability of LC-MS coupled with bioinformatic analysis to detect carryover where CE with UV detection was not able to detect any carryover was demonstrated. The findings from this approach suggest that the acceptance of chromatographic resin reuse for multiple products will be dependent on the development of analytical/bioinformatics tools such as those proposed in this work. [Display omitted] • A multiple product Protein A resin reuse process was designed. • Capillary electrophoresis with UV detection was not able to detect any carryover. • LC-MS coupled with bioinformatic analysis did detect carryover. • These data provide leverage for chromatographic resin reuse for biopharmaceuticals. [ABSTRACT FROM AUTHOR]

Internal and external quality assurance materials often use highly processed matrixes. This can render the materials non-commutable. Monitoring laboratory methods with patient medians helps in identifying and correcting systematic errors that may affect diagnostic accuracy. The aim of the present study was to use HbA1c patient results for monitoring of method performance over time. Test HbA1c results from 2010 to 2022 was analyzed (n=722,553) regarding changes over time and seasonal variation. The HbA1c testing was initially performed on a Cobas 501 instrument using immunological detection but in May 2017 the method was replaced by capillary electrophoresis on Capillarys 3 Tera. There was a steady decrease in HbA1c values. From 2011 to 2021 the decrease was for 0.10 percentile 6.6 %, lower quartile 7.9 %, median 10.2 %, mean values 9 %, upper quartile 11.2 %, and 0.90 percentile 9.3 %. No clear shift in HbA1c levels was observed due to the shift in methods. The median HbA1c values per month was approximately 44 mmol/mol (6.2 %, DCCT/NGSP). The only month with a median HbA1c that differed by more than 1 mmol/mol was July with a median value of 42 mmol/mol (6.0 %). The patient data showed a similar decrease as in the National Diabetes Register which indicates that the method is stable over time without any sudden changes and that the seasonal variation is low. The continuous decrease in HbA1c values over time is most likely to a shift towards earlier detection of patient with diabetes and improved treatment. [ABSTRACT FROM AUTHOR]

Cerebellar ataxia, neuropathy, and vestibular areflexia syndrome (CANVAS) is caused by biallelic pathogenic expansions, or compound heterozygosity with other pathogenic variants in the RFC1 gene. CANVAS is estimated to be underdiagnosed, both because of the lack of formal diagnostic criteria and molecular challenges that translate to lesser access and high cost of routine testing. Our aim was to address the need for making CANVAS genetic testing routine, by designing a streamlined two‐step PCR consisting of a short‐allele screening PCR and a confirmatory PCR with fragment capillary electrophoresis detection. Exome sequencing of RFC1 was additionally foreseen to resolve potential compound heterozygosity cases. Specificity of our approach was evaluated using ataxia patients with known non‐CANVAS diagnoses, and optimized using Southern blot confirmed CANVAS patients. We evaluated our approach by testing patients consecutively referred for clinically suspected CANVAS using first the two‐step PCR, followed by exome sequencing. Our approach was able to accurately identify negative and confirm positive cases in prospectively collected suspected CANVAS patients presenting with at least three typical clinical signs. The proposed testing approach provides an alternative method able to clearly distinguish between CANVAS negative and positive cases and can be easily incorporated into the genetic diagnostic laboratory workflow. [ABSTRACT FROM AUTHOR]

AbstractStatins are the most prescribed drug for regulating the high cholesterol level in the blood, which can lead to severe complications, such as cardiovascular diseases and other health complications. A wide range of analytical techniques have been employed for the quantification of statins from various origins, including fermentation derived (lovastatin, pravastatin, and compactin), semi-synthetic (simvastatin), and synthetic (atorvastatin, rosuvastatin, and fluvastatin) routes. The presence of more than one structural form and structural analogue generated in the biosynthesis pathway, as well as reaction intermediates and macromolecules in the clinical sample, complicates the quantification of statins. Furthermore, significant concentrations of statins in environmental samples pose serious health and ecology hazards, and estimating statins in those diluted samples is extremely difficult. On the other hand, the: cost, accurate estimation of the desired one from other structural forms, sample complexity, time, limits of detection and quantification, were major criteria distinguishing the usability of each technique. As a result, the current manuscript focuses on analytical techniques such as molecular spectroscopy (normal and derivatives UV-Visible spectrophotometer), chromatography (TLC, HP-TLC, HPLC, GC, swing column, micellar, and supercritical fluid), mass spectroscopy (HPLC-MS/MS and GC-MS/MS), sequential flow injection, capillary electrophoresis, and cyclic voltammetry, as well as their: optimal operating conditions, limits of detection and quantification, advancements, and limitations. Furthermore, various online and offline sample preparations (precipitation, solid phase extraction, liquid-liquid extraction, and micellar extraction) have been highlighted as an essential pretreatment technique to avoid the interference caused by structural analogues and other macromolecules. The greener and more sustainable concepts used in analytical approaches for the quantification statins are also highlighted with current advancements. [ABSTRACT FROM AUTHOR]

Globally, intron 22 inversions (Inv22s) of the factor VIII gene (F8) are the most frequent pathogenic variants and account for 45–50% of severe hemophilia A (SHA) cases, while intron 1 inversion (Inv1) explains 1–5% of SHA cases. The detection of both inversions by an inverse shifting-polymerase chain reaction (IS-PCR) is the first choice worldwide for the diagnosis of patients and carriers of SHA. To improve its sensitivity and reproducibility in the visualization of PCR products, we approached the IS-PCR with fluorescent capillary electrophoresis instead of agarose electrophoresis. Based on the original protocol, we modified two primers by 5'-end labeling with FAM™ fluorescent dye for the detection of the PCR products by capillary electrophoresis. Additionally, the "fast enzymes" BclI and T4-Ligase were incorporated for work saving in the genomic digestion and ligation reactions, respectively. Once we accomplished the standardization and verified the reproducibility of the modified IS-PCR method, we applied it for the diagnosis of a cohort of SHA patients and carriers. The modified IS-PCR by fluorescent capillary electrophoresis for PCR product detection is more sensitive than agarose electrophoresis. The method was also improved by using the new rapid enzymes to save time in the whole process. [ABSTRACT FROM AUTHOR]

The current review provides a focused analysis of the application of capillary electrophoresis (CE) techniques to determine the chiral purity of pharmaceuticals, with a specific emphasis on cyclodextrin- (CD) based chiral selectors (CSs), highlighting advancements, methodologies, and trends in this area as reported in studies published from 2010 to 2024. The review emphasizes CE's evolution as a critical tool in this field, discussing its advantages, such as high efficiency, flexibility, relatively low costs, and minimal environmental impact, which make it well-suited for modern pharmaceutical applications. Additionally, it underscores the importance of CE in meeting stringent regulatory requirements for chiral drug substances. A significant shift in method optimization has occurred in the last ten years, shifting from the traditional One-Factor-at-a-Time (OFAT) strategy to the Design-of-Experiments (DoE) approach; this shift has enabled more systematic and robust method development. Furthermore, a common trend in recent years is the application of Quality-by-Design (QbD) principles in method development and optimization, ensuring higher reliability and efficiency. Additionally, there is an increasing focus on developing CE methods capable of detecting both achiral and chiral impurities simultaneously, which enhances the comprehensiveness of the analysis. This review seeks to guide future research and development in optimizing CE methodologies for pharmaceutical applications. [ABSTRACT FROM AUTHOR]

Short‐chain fatty acids (SCFAs) are essential molecules produced by gut bacteria that fuel intestinal cells and may also influence overall health. An imbalance of SCFAs can result in various acute and chronic diseases, including diabetes, obesity and colorectal cancer (CRC). This review delves into the multifaceted roles of SCFAs, including a brief discussion on their source and various gut‐residing bacteria. Primary techniques used for detection of SCFAs, including gas chromatography, high‐performance gas chromatography, nuclear magnetic resonance and capillary electrophoresis are also discussed through this article. This review study also compiles various synthesis pathways of SCFAs from diverse substrates such as sugar, acetone, ethanol and amino acids. The different pathways through which SCFAs enter cells for immune response regulation are also highlighted. A major emphasis is the discussion on diseases associated with SCFA dysregulation, such as anaemia, brain development, CRC, depression, obesity and diabetes. This includes exploring the relationship between SCFA levels across ethnicities and their connection with blood pressure and CRC. In conclusion, this review highlights the critical role of SCFAs in maintaining gut health and their implications in various diseases, emphasizing the need for further research on SCFA detection, synthesis and their potential as diagnostic biomarkers. Future studies of SCFAs will pave the way for the development of novel diagnostic tools and therapeutic strategies for optimizing gut health and preventing diseases associated with SCFA dysregulation. Significance Statement: Understanding the role of short‐chain fatty acids (SCFAs) is crucial because they are fundamental to maintaining colonic health and overall bodily function. Imbalances in SCFAs are linked to serious conditions like diabetes, obesity and colorectal cancer, making it essential to explore their regulation and impact. Additionally, studying the variations in SCFA levels across different ethnicities can provide insights into disease susceptibility and health disparities. Furthermore, advancing detection methods and developing sensors for SCFAs could revolutionize diagnostics, allowing for earlier and more precise intervention in metabolic and gut‐related diseases. Thus, this review addresses significant gaps in knowledge and technology that could lead to better health outcomes. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides take a specific position in the field of antibiotics (ATBs), however, from a large number of available molecules only a few of them were approved and are used in clinics. These therapeutic modalities play a crucial role in the management of diseases caused by multidrug‐resistant bacterial pathogens and represent the last‐line therapy for bacterial infections. Therefore, there is a demand for a rationale use of such ATBs based on optimization of the dosing strategy to minimize the risk of resistance and ensure the sustainable efficacy of the drug in real clinical practice. Therapeutic drug monitoring, as a measurement of drug concentration in the body fluids or tissues, results in the optimization of the patient´s medication and therapy outcome. This strategy is beneficial and could result in tailored therapy for different types of infection and the prolongation of the use and efficacy of ATBs in hospitals. This review paper provides an actual overview of approved antimicrobial peptides used in clinical practice and covers current trends in their analysis by convenient and advanced methodologies used for their identification and/or quantitation in biological matrices for therapeutic drug monitoring purposes. Special emphasis is given to the methods with perspective clinical outcomes. [ABSTRACT FROM AUTHOR]

The increasing awareness of environmental issues and the transition to green analytical chemistry (GAC) have gained popularity among academia and industry in recent years. One of the principles of GAC is the reduction and replacement of toxic solvents with more sustainable and environmentally friendly ones. This review gives an overview of the advances in applying green solvents as an alternative to the traditional organic solvents for peptide and protein purification and analysis by liquid chromatography (LC) and capillary electrophoresis (CE) methods. The feasibility of using greener LC and CE methods is demonstrated through several application examples; however, there is still plenty of room for new developments to fully realize their potential and to address existing challenges. Thanks to the tunable properties of designer solvents, such as ionic liquids and deep eutectic solvents, and almost infinite possible mixtures of components for their production, it is possible that some new designer solvents could potentially surpass the traditional harmful solvents in the future. Therefore, future research should focus mainly on developing new solvent combinations and enhancing analytical instruments to be able to effectively work with green solvents. [ABSTRACT FROM AUTHOR]

In this work, capillary electrophoresis with Gaussian distribution‐based peak fitting for improved peak resolution for simultaneous quantitative analysis of chondroitin sulfate (CS) and dermatan sulfate (DS) was developed for the first time. The effects of different pH, voltage, and peak shapes on the peak fitting were investigated. The results indicate that separation conditions that affect peak shape have a significant impact on peak fitting. Under optimized separation conditions, the peak fitting achieves high accuracy and precision, the relative standard deviation (n = 5) of peak area and migration time of CS and DS were 1.1%, 1.8% and 0.94%, 0.84%, respectively. Furthermore, the results obtained by peak fitting were close to real values. The developed method was used to analyze the purity of CS. The results had a high consistency with the labeled value; furthermore, the peak fitting could point out the number of possible impurities. The developed method has good potential for the analysis of other glycosaminoglycans with peak overlap. [ABSTRACT FROM AUTHOR]

Background: Laboratory rats, as model animals, have been extensively used in the fields of life science and medicine. It is crucial to routinely monitor the genetic background of laboratory rats. The conventional approach relies on gel electrophoresis and capillary electrophoresis (CE) technologies. However, the experimental and data analysis procedures for both of these methods are time consuming and costly. Results: We established a single-nucleotide polymorphism (SNP) typing scheme using multiplex polymerase chain reaction (PCR) and next-generation sequencing (NGS) to address the genetic background ambiguity in laboratory rats. This methodology involved three rounds of PCR and two rounds of magnetic bead selection to improve the quality of the sequencing data. We simultaneously analysed 100 laboratory rats (including rats of 5 inbred strains and 2 in-house closed colonies), and the sequencing depth varied from an average of 108.25 to 5189.89, with sample uniformity ranging from 82.5 to 97.5%. A total of 98.9% of the amplicons were successfully genotyped (≥ 30 reads). Genetic background analysis revealed that all 38 experimental rats from the 5 inbred strains were successfully identified (without a heterozygous allele). For the 2 in-house closed colonies, the average heterozygosity (0.162 and 0.169) deviated from the typical range of 0.5–0.7, indicating a departure from the ideal heterozygosity level. Additionally, we employed multiplex PCR-CE to validate the NGS-based method, which yielded consistent results for all the rat strains. These results demonstrated that this approach significantly improves efficiency, saves time, reduces costs and ensures accuracy. Conclusion: By utilizing NGS technology, our developed method leverages SNP genotyping for genetic background identification in laboratory rats, demonstrating advantages in terms of labour efficiency and cost-effectiveness, thereby rendering it well suited for projects involving extensive sample cohorts. [ABSTRACT FROM AUTHOR]

Context.--Existing targeted cystic fibrosis screening assays miss important pathogenic CFTR variants in the ethnically diverse US population. Objective.--To evaluate the analytic performance of a multiplex polymerase chain reaction (PCR)/capillary electropho-resis (CE) CFTR assay panel that simultaneously interrogates primary pathogenic variants of different ethnic/ancestral groups. Design.--Performance characteristic assessment and variant coverage comparison of the panel with a focus on ethnicity-specific CFTR variants were performed. Sample DNA was primarily from whole blood or cell lines. Detection of CFTR carriers was compared across several commercially available CFTR kits and recommended variant sets based on panel content. Results.--The panel interrogated 65 pathogenic CFTR variants representing 92% coverage from a recent genomic sequencing survey of the US population, including 4 variants with top 5 frequency in African or Asian populations not reflected in other targeted panels. In simulation studies, the panel represented 95% of carriers across the global population, resulting in a 6.9% to 19.0% higher carrier detection rate compared with 10 targeted panels or variant sets. Precision and sensitivity/specificity were 100% concordant. Multisite sample-level genotyping accuracy was 99.2%. Across PCR and CE instruments, sample-level genotyping accuracy was 97.1 %, with greater than 99% agreement for all variant-level metrics. Conclusions.--The CFTR assay achieves 92% or higher coverage of CFTR variants in diverse populations and provides improved pan-ethnic coverage of minority subgroups of the US populace. The assay can be completed within 5 hours from DNA sample to genotype, and performance data exceed acceptance criteria for analytic metrics. This assay panel content may help address gaps in ancestry-specific CFTR genotypes while providing a streamlined procedure with rapidly generated results. [ABSTRACT FROM AUTHOR]

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A moment analysis method was applied to study intermolecular interactions between phenylboronic acid and monosaccharides. The values of first absolute and second central moments of elution peaks, measured by affinity capillary electrophoresis (ACE), were analyzed on the basis of the moment theory to determine those of association equilibrium constant and association and dissociation rate constants of the intermolecular interactions. Their values were fairly in agreement with results reported in previous studies. The moment analysis method using ACE has some advantages in comparison with conventional spectroscopic methods. For example, neither immobilization nor fluorescence labelling of solute and/or ligand molecules is required when ACE data are measured. Because the principles of the method are different from those of ordinary spectroscopic ones, this method contributes to the kinetic study of intermolecular interactions from various perspectives. A comparison was made between intrinsic characteristics of the moment analysis method and those of two historical ones developed by the Whitesides and Krylov groups. Some restrictive features of the moment analysis method were also explained. It is concluded that the moment analysis method using ACE is effective as one experimental method for the analysis of intermolecular interactions. [ABSTRACT FROM AUTHOR]

This paper describes the use of PolyJet 3D printing to fabricate microchip electrophoresis devices with integrated microwire electrodes for amperometric detection. The fabrication process involves 3D printing of two separate pieces, a channel layer and an electrode layer. The channel layer is created by 3D printing on a pre-fabricated mold with a T-intersection. For the electrode layer, a stencil design is printed directly on the printing tray and covered with a piece of transparent glass. Microwire electrodes are adhered over the glass piece (guided by underlaying stencil) and a CAD design of the electrode layer is then printed on top of the microwire electrode. After delamination from the glass after printing, the microwire is embedded in the printed piece, with the stencil design ensuring that alignment and positioning of the electrode is reproducible for each print. After a thermal bonding step between the channel layer and electrode layer, a complete electrophoresis device with integrated microelectrodes for amperometric detection results. It is shown that this approach enables different microwire electrodes (gold or platinum) and sizes (100 or 50 µm) to be integrated in an end-channel configuration with no gap between the electrode and the separation channel. These devices were used to separate a mixture of catecholamines and the effect of separation voltage on the potential voltage applied on the working electrode was also investigated. In addition, the effect of electrode size on the number of theoretical plates and limit of detection was studied. Finally, a device that contains different channel heights and a detection electrode was 3D-printed to integrate continuous flow sampling with microchip electrophoresis and amperometric detection. [ABSTRACT FROM AUTHOR]

The significance and necessity of separating enantiomers in food, pharmaceuticals, pesticides, and other samples remains constant and unrelenting. The successful chiral separation usually includes the application of a chiral auxiliary compound, known also as a chiral selector (CS), that forms complexes with enantiomers of different physicochemical properties, enabling efficient separation. While both native and substituted cyclodextrins (CDs) are commonly used as CSs, β-CD is undoubtedly the most popular one among them. This review includes recent advancements in the application of β-CD as a CS. While the theoretical background behind the enantioseparation is also part of this work, the main emphasis is put on the factors that affect the efficacy of this process such as temperature, pH, solvent, and the choice of other additives. Also, the different analytical methods: Nuclear Magnetic Resonance (NMR) spectroscopy, Capillary Electrophoresis (CE), fluorescence spectroscopy (FS), High-Performance Liquid Chromatography (HPLC), Isothermal Titration Calorimetry (ITC), and UV–vis spectroscopy, used for enantioseparation with the aid of β-CD as CS, are thoroughly compared. Also, since some of the chiral compounds have been studied in the context of their enantioseparation more than once, those works are compared and critically analyzed. In conclusion, while β-CD can be in most cases used as CS, the choice of the experimental conditions and method of analysis is crucial to achieve the success. [ABSTRACT FROM AUTHOR]

Multiple‐step on‐line preconcentration, a combination of at least two stacking techniques has been developed to increase the sensitivity in capillary electrophoresis (CE) for analytes in various samples. It is usually conducted sequentially, or in some cases, synergistically, where different stacking modes occur simultaneously. Multiple‐step techniques allow simultaneous preconcentration and separation of various kinds of analytes in different complex samples in a single CE run. This review aims to provide recent advances in multiple‐step on‐line preconcentration techniques in CE. We critically review technical papers published for the last 7 years up until July 2024, subsequently organized according to the combination of the main stacking techniques, that is, field amplification, large volume sample stacking, transient isotachophoresis, micelle to solvent or micelle to cyclodextrin stacking, and others. The procedures, fundamental mechanism, analytical figures of merits achieved, and their feasibility for complicated sample matrices are reviewed. [ABSTRACT FROM AUTHOR]

In forensic genetics, utilizing massively parallel sequencing (MPS) to analyze short tandem repeats (STRs) has demonstrated several advantages compared to conventional capillary electrophoresis (CE). Due to the current technical limitations, although flanking region polymorphisms had been mentioned in several previous studies, most studies focused on the core repeat regions of STRs or the variations in the adjacent flanking regions. In this study, we developed an MPS system consisting of two sets of multiplex PCR systems to detect not only the STR core repeat regions but also to observe variants located at relatively distant positions in the flanking regions. The system contained 42 commonly used forensic STRs, including 21 autosomal STRs (A-STRs) and 21 Y-chromosomal STRs (Y-STRs), and a total of 350 male individuals from a Chinese Han population were genotyped. The length and sequence variants per locus were tallied and categorized based on length (length-based, LB), sequence without flanking region (core repeat regions sequence-based, RSB), and sequence with flanking region (core repeat and flanking regions sequence-based, FSB), respectively. Allele frequencies, Y-haplotype frequencies, and forensic parameters were calculated based on LB, RSB, and FSB, respectively, to evaluate the improvement in discrimination power, heterozygosity, and effectiveness of forensic systems. The results suggested the sequence variations have more influence on A-STRs and could improve the identification ability of MPS-STR genotyping. Concordance between MPS and CE methods was confirmed by using commercial CE-based STR kits. The impact of flanking region variations on STR genotype analysis and potential factors contributing to discordances were discussed. A total of 58 variations in the flanking regions (53 SNPs/SNVs and 5 InDels) were observed and most variations (48/58) were distributed in A-STRs. In summary, this study delved deeper into the genetic information of forensic commonly used STR and advanced the application of massively parallel sequencing in forensic genetics. [ABSTRACT FROM AUTHOR]

【Objective】 Genetic diversity analysis and evaluation of introduced potato varieties is a necessary method for screening good parental materials. 【Method】SSR fluorescent marker capillary electrophoresis was used to analyze the genetic diversity of 51 introduced potato varieties and construct molecular ID cards. 【Result】The 41 pairs of primers amplified a total of 327 alleles, and 311 alleles were differentiated at the same position, and the percentage of polymorphism was 95.11%. The primer polymorphism information index (PIC) ranged from 0 to 0.89, with an average of 0.71. Shannon's (I) ranged from 0 to 0.54, with an average of 0.32. The genetic similarity coefficient (GS) of the introduced potato varieties was 0.32. In the genetic similarity coefficient GS of 0.76, the test materials were categorized into six major groups: Group I having 14 varieties, group II having 30 varieties, group III having three varieties, group V having two varieties, and group IV and VI having one variety each. Cluster analysis revealed that four varieties, Vilas, Dingshu 4, Kexin 33 and Dongnong 312 could be used as potato breeding materials. 【Conclusion】Three pairs of core primers, STM0037, SSR0387 and SSR01054, were selected to construct fingerprints and molecular IDs of 51 potato varieties. [ABSTRACT FROM AUTHOR]

A fast, simple and cost-effective capillary electrophoresis (CE) method was developed for caffeine determination in different beverages (energy drinks, soft drinks). Because caffeine is neutral from electrophoretic point of view and migrates with the electroosmotic flow (EOF), micellar electrokinetic chromatography (MEKC) was chosen as the separation method. The optimum separation conditions consisted of 25 mM sodium tetraborate, 100 mM sodium dodecyl sulphate, pH 9.30, 20°C temperature, 20 kV voltage, 50 mbar/sec hydrodynamic injection, UV detection at 270 nm. Employing the optimized conditions caffeine was quantified in less than 3 minutes. The analytical performances of the method were verified in terms of accuracy, linearity, limit of detection and quantification, precision and robustness. The method was applied also to detect caffeine in coffee and tea. The advantage of MEKC over other analytical methods, particularly compared with the more frequently used HPLC methods, lies in its lower operating costs and higher environmental friendliness. [ABSTRACT FROM AUTHOR]

A simple, fast, and low-cost pre-concentration methodology based on the application of solid-phase extraction coupled to layered double hydroxides (LDHs) and capillary electrophoresis was developed for the determination of naproxen (NPX), diclofenac (DFC), and ibuprofen (IBP) in environmental sample waters. A systematic study of the LDH composition was designed, including the effects of interlayer anions (NO3−, Cl−, CO32−, BenO−, and SDS−) and the effect of molar ratio (Mg:Al). The optimal composition of MgAl/Cl−-LDH (Mg:Al; 1.5:1.0) was coupled to an SPE system: pH (neutral pH), LDH amount (15 mg), and extraction capacity ranged from 79.71 to 83.11% for the three anti-inflammatory non-steroidal agents analyzed. A recovery rate of up to 80.87% was obtained when 0.01 M chloride acid in methanol was used as the eluent and 50 mL of sample was used. Under optimal conditions, the linear range of the calibration curve ranges from 18.02 to 200 μg L−1, with limits of detection ranging from 6.03 to 18.02 μg L−1 for the three NSAIDs. The precision of the methodology was evaluated in terms of inter- and intra-day repeatability, with %RSD < 10% in all cases. The proposed method was applied to analyze environmental water samples (bottle, tap, cistern, well, and river water samples). The developed method is a robust technique capable of combining with other analytical methods to quantitatively determine anti-inflammatory non-steroidal agents. [ABSTRACT FROM AUTHOR]

In recent years, the consumption of dietary supplements, particularly those incorporating plant-based ingredients, has increased greatly, driven by the perception of their natural origins and purported minimal health risks. However, one significant safety concern revolves around the adulteration of dietary supplements, wherein unscrupulous manufacturers may illegally incorporate pharmaceutical substances or their analogs into these products to achieve increased efficiency and bolster sales. This review assesses the role of capillary electrophoresis (CE) in ensuring the quality control of dietary supplement products over the past two decades. This study provides an overview of various applications of CE in analyzing dietary supplements, outlining the typical attributes of natural product analysis using CE. These analyses demonstrate the broad versatility of CE, exemplified by its diverse applications and detection modes. Moreover, the review highlights the growing prominence of CE as a separation technique in quality control, by comparison with more conventional methods like high-performance liquid chromatography (HPLC). Through this exploration, the review elucidates the pivotal role of CE in upholding the integrity and safety of dietary supplements, in connection with a landscape of evolving regulatory challenges and consumer demands. [ABSTRACT FROM AUTHOR]

HbA1c is a pivotal biomarker in diabetes management, reflecting long-term glycaemic control. HbA1c is often measured with capillary electrophoresis, which generally is a very precise technique, but there can be interference, especially in the case of haemoglobin diseases. Thus, in patients with underlying conditions, the accurate measurement of HbA1c can be challenging. We present a case of special interference in a 74-year-old female patient referred to a HbA1c test, in whom the measurement was thought to be significantly affected by hyperleukocytosis and led to an unexpected diagnosis of leukemic low-grade lymphoma. This case report highlights the underrecognized potential interference of leukocytosis in HbA1c measurement. It underscores the importance of clinical vigilance when interpreting HbA1c results in patients with underlying haematological conditions. [ABSTRACT FROM AUTHOR]

Background/Objectives: The objective of this study was to investigate the metabolomic profiles of patients with colorectal cancer (CRC) across various stages of the disease. Methods: The plasma samples were obtained from 255 subjects, including patients with CRC in stages I-IV, polyps, and controls. We employed capillary electrophoresis time-of-flight mass spectrometry and liquid chromatography triple quadrupole mass spectrometry to analyze hydrophilic metabolites comprehensively. The data were randomly divided into two groups, and consistent differences observed in both groups were analyzed. Results: Acetylated polyamines, such as N1-acetylspermine and N1, N12-diacetylspermine, consistently showed elevated concentrations in stage IV compared to stages I-III. Non-acetylated polyamines, including spermine and spermidine, exhibited increasing trends from polyp to stage IV. Other metabolites, such as histidine and o-acetylcarnitine, showed decreasing trends across stages. While acetylated polyamines have been reported as CRC detection markers, our findings suggest that they also possess diagnostic potential for distinguishing stage IV from other stages. Conclusions: This study showed stage-specific changes in metabolic profiles, including polyamines, of colorectal cancer. [ABSTRACT FROM AUTHOR]

Background Hemoglobin (Hb) Hekinan II (A27; Glu-Asp) is an α-chain variant, and its interaction with the common Southeast Asian (--SEA/) α-thalassemia (α-thal) deletion is rarely reported. This study provides a clinical update of Hb Hekinan II associated with (--SEA/) α-thal. Methods A total of 11 simple heterozygotes and 20 composite heterozygotes for Hb Hekinan II and (--SEA/) α-thal were included based on molecular diagnosis. Results Hb Hekinan II exhibited a significant increase in hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin content, but a decrease in red blood cell level compared with α+ thalassemia deletion. Compared with (--SEA/) α-thal, composite heterozygotes for Hb Hekinan II and (--SEA/) α-thal showed similar erythrocyte parameters. Both heterozygotes with and without (--SEA/) α-thal showed low Hb A2 level. Hb Hekinan II showed abnormal performance in high-performance liquid chromatography but not in capillary electrophoresis. Conclusion Hb Hekinan II is a benign Hb variant. The heterozygotes exhibit clinically asymptomatic coinheritance with (--SEA/) α-thal having comparable hematological phenotype to simple (--SEA/) α-thal. The combination of hematological and molecular analysis helped to improve the detection rate of this rare variant. [ABSTRACT FROM AUTHOR]

Objectives Voxelotor can increase hemoglobin levels in patients living with sickle cell disease (SCD). A clinician who is monitoring voxelotor response may want to know whole-blood voxelotor concentration, but this cannot be measured in most clinical settings. However, voxelotor has been demonstrated to cause "peak splitting" in common methods of hemoglobin measurement such as capillary zone electrophoresis (CZE) and high-performance liquid chromatography (HPLC). We hypothesized that we could use the size of the peak split to estimate the whole-blood concentration. Methods Blood from people with SCD was dosed with known concentrations of voxelotor, and multiparameter regression was used to derive the relationship of voxelotor concentration to the degree of peak splitting observed. To validate these equations, 21 patients started on voxelotor at 1500 mg/d had blood samples drawn at days 0, 14, 30, and 60. Samples were sent out for gold standard voxelotor concentration testing. The derived equations were then used to calculate voxelotor concentration. Results Calculated concentrations correlated strongly with measured concentrations for both CZE (R 2 = 0.83, P <.001) and HPLC (R 2 = 0.76, P <.001). Voxelotor concentration also had a significant effect on increases in hemoglobin (R 2 = 0.40, P <.001). Conclusions Thus, peak splitting CZE and HPLC can be used to estimate voxelotor concentration. [ABSTRACT FROM AUTHOR]

Sulfonylurea herbicides (SUHs) are widely used in agriculture because of their low dosage, low cost, and high selectivity. However, due to improper use and lack of effective management, their residues pose a threat to the human health through environment and food pollution. Therefore, there is a need for simple, quick, economical, and effective methods to analyze SUHs in plant-derived foods, crops, and environmental samples. The present article presents a comprehensive review of the pretreatment and analytical technologies used for SUHs in various sample matrices, focusing on the developments since 2010. The main pretreatment methods include liquid–liquid extraction, solid-phase extraction, QuEChERS, and different microextraction methods, whereas analytical methods mainly include liquid chromatography coupled with different detectors, capillary electrophoresis, among others. In addition, the present study also compared the advantages and disadvantages of the methods and the future development is prospected. Highlight: Progress in pretreatment and analytical methods of sulfonylurea herbicides are summarized. Pros and cons of the pretreatment and determination methods have been given. Novel materials in solid-phase extraction and solid-phase microextraction are depicted. [ABSTRACT FROM AUTHOR]

Commercial short tandem repeat (STR) kits exclusively contain human-specific primers; however, various non-human organisms with high homology to the STR kit's primer sequences can cause cross-reactivity. Owing to the proprietary nature of the primers in STR kits, the origins and sequences of most non-specific peaks (NSPs) remain unclear. Such NSPs can complicate data interpretation between the casework and reference samples; thus, we developed "NSPlex", an efficient method to discover the biological origins of NSPs. We used leftover STR kit amplicons after capillary electrophoresis and performed advanced bioinformatics analyses using next-generation sequencing followed by BLAST nucleotide searches. Using our method, we could successfully identify NSP generated from PCR amplicons of a sample mixture of human DNA and DNA extracted from matcha powder (finely ground powder of green tea leaves and previously known as a potential source of NSP). Our results showed our method is efficient for NSP analysis without the need for the primer information as in commercial STR kits. [ABSTRACT FROM AUTHOR]

Electroosmotic flow significantly impacts the resolution of separations in capillary electrophoresis and its modification is often necessary. Coating of the inner capillary surface either dynamically or permanently offers a way to alter the electroosmotic flow, potentially reducing the adsorption of analytes, in our case liposomes, to the capillary wall. At first, we measured fluorescently labeled liposomes in an uncoated capillary by capillary electrophoresis with laser-induced fluorescence detection. We used a special procedure of the electrophoretic experiment allowing us to observe the development of peak shape at the early stages of migration. We proved that the liposomes were adsorbed to the capillary wall, which led to a very quick and severe dispersion of their peak during their electromigration. For this reason, we used a commercially coated capillary with polyvinyl alcohol, where at the same separation conditions, we observed the peak of the liposome with a stable shape during the migration. However, this capillary is costly, thus four simple dynamic coating methods were tested for four polymers, namely Pluronic F-127, polyvinyl pyrrolidone K30, polyethylene glycol, and polydiallyldimethylammonium chloride. Among them, we chose a method where we flushed the capillary with a 10% solution of polyvinyl pyrrolidone K30 before the first measurement. In addition, in-house made permanent coating with linear polyacrylamide was investigated, which has led to effective suppression of the electroosmotic flow and stable liposome peak, not dispersed during its migration. Liposome separation using this coating fully confirmed that the studied liposomes are negatively charged and migrate in the anodic direction. [ABSTRACT FROM AUTHOR]

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The present study aimed to conduct a comparative investigation of the biological properties of phenolic and polysaccharide extracts obtained using an ultrasound-assisted technique from Aloe vera gel and their effects on each stage of the wound healing process in in vitro experimental models. HPLC analysis showed that the phenolic extract contained aloin, ferulic, and caffeic acid, as well as quercetin dihydrate, as major compounds. Capillary zone electrophoresis indicated the prevalence of mannose and glucose in the polysaccharide extract. Cell culture testing revealed the anti-inflammatory properties of the phenolic extract at a concentration of 0.25 mg/mL through significant inhibition of pro-inflammatory cytokines—up to 28% TNF-α and 11% IL-8 secretion—in inflamed THP-1-derived macrophages, while a pro-inflammatory effect was observed at 0.5 mg/mL. The phenolic extract induced 18% stimulation of L929 fibroblast proliferation at a concentration of 0.5 mg/mL, enhanced the cell migration rate by 20%, and increased collagen type I synthesis by 18%. Moreover, the phenolic extract exhibited superior antioxidant properties by scavenging free DPPH (IC50 of 2.50 mg/mL) and ABTS (16.47 mM TE/g) radicals, and 46% inhibition of intracellular reactive oxygen species (ROS) production was achieved. The polysaccharide extract demonstrated a greater increase in collagen synthesis up to 25%, as well as antibacterial activity against Staphylococcus aureus with a bacteriostatic effect at 25 mg/mL and a bactericidal one at 50 mg/mL. All these findings indicate that the phenolic extract might be more beneficial in formulations intended for the initial phases of wound healing, such as inflammation and proliferation, while the polysaccharide extract could be more suitable for use during the remodeling stage. Moreover, they might be combined with other biomaterials, acting as efficient dressings with anti-inflammatory, antioxidant, and antibacterial properties for rapid recovery of chronic wounds. [ABSTRACT FROM AUTHOR]

Background: Application of electromembrane extraction coupled with capillary electrophoresis was studied for ofloxacin extraction from plasma samples. Methods: Ofloxacin migrated from acidic plasma samples through a thin layer of 1-octanol immobilized in the pores of a porous hollow fiber wall into a 10 µL acidic aqueous acceptor solution located inside the lumen of the fiber. Results: Under optimum conditions influencing electromigration (i.e., 20 min of the operation time, stirring speed of 750 rpm, donor phase pH at 4.0, acceptor pH at 3.0, and applied voltage of 30 V across the supported liquid membrane), ofloxacin was extracted from plasma samples with an enrichment factor of 100-fold corresponding to extraction percent of 24%. The calibration curve showed acceptable linearity in the range of 0.2-7.0 µg.mL-1 (R=0.9993). The limits of detection and quantification of 0.05 and 0.2 µg.mL-1 were obtained, respectively. The inter- and intra-day precision and accuracy were obtained below 8.65%. Conclusion: The validated method was successfully processed for the ofloxacin determination in the plasma samples of patients under ofloxacin therapy. There were no interfering peaks which indicates the great selectivity of the developed method.

N-glycosylation can have a profound effect on the quality of mAb therapeutics. In biomanufacturing, one of the ways to influence N-glycosylation patterns is by altering the media used to grow mAb cell expression systems. Here, we explore the potential of machine learning (ML) to forecast the abundances of N-glycan types based on variables related to the growth media. The ML models exploit a dataset consisting of detailed glycomic characterisation of Anti-HER fed-batch bioreactor cell cultures measured daily under 12 different culture conditions, such as changes in levels of dissolved oxygen, pH, temperature, and the use of two different commercially available media. By performing spent media quantitation and subsequent calculation of pseudo cell consumption rates (termed media markers) as inputs to the ML model, we were able to demonstrate a small subset of media markers (18 selected out of 167 mass spectrometry peaks) in a Chinese Hamster Ovary (CHO) cell cultures are important to model N-glycan relative abundances (Regression - correlations between 0.80–0.92; Classification - AUC between 75.0–97.2). The performances suggest the ML models can infer N-glycan critical quality attributes from extracellular media as a proxy. Given its accuracy, we envisage its potential applications in biomaufactucuring, especially in areas of process development, downstream and upstream bioprocessing.

An in-line sample concentration method for capillary electrophoresis called admicelle to solvent microextraction was proposed. In this technique, analytes were trapped in the cetyltrimethylammonium bromide admicelles formed in situ on the negatively charged capillary surface. A solvent plug was then partially injected hydrodynamically to collapse the admicelles, which liberated and focused the analytes at the solvent front. Voltage was applied across the capillary, completing the stacking process. Various solvents, namely, methanol, ethanol, and acetonitrile, were investigated. The optimal solvent for solvent to admicelle microextraction was 30% acetonitrile in 24 mM sodium tetraborate (pH 9.2). Sample injection time and solvent to sample injection ratio were also optimised. For this demonstration, the optimum sample injection time and solvent to sample injection ratio were 320 s and 1:2, respectively. Using the optimum conditions, UV detection sensitivity was enhanced 132–176-fold for the model anions. The LOQ, %intra-/inter-day (n = 6/n = 12, 2 days) repeatability, and linearity (R2) of admicelle to solvent microextraction were 0.08–2 µg/mL, 1.9–3.9%, 2.8–4.9%, and 0.992, respectively. Admicelle to solvent microextraction was applied to the analysis of various fortified water samples, with good repeatability (%RSD = 0.5–3.6%), and no matrix interferences. [ABSTRACT FROM AUTHOR]

Genetically modified (GM) crops and their products are regulated in many countries. To meet increasing regulatory requirements for a growing number of GM soybean events, high-throughput and cost-effective detection methods are indispensable. In this study, 14 widely commercialized GM soybean events, including GTS40-3-2, MON89788, CV127, A2704-12, A5547-127, 305423, 356043, MON87701, MON87705, MON87708, MON87769, SYHT0H2, FG72, and DAS-444Ø6-6 were utilized as targets for event-specific identification. A high-throughput detection method was established by integrating a single universal primer multiplex PCR (SUP-M-PCR) with capillary electrophoresis technique. This method enables the identification of 14 soybean events in a single PCR reaction, addressing the limitations of conventional multiplex PCR such as self-inhibition and amplification variations between different primers. Furthermore, it significantly enhances the sensitivity and accuracy of previous multiple detection methods, achieving a detection sensitivity of 0.05% (w/w). Overall, this approach greatly improves detection efficiency and holds promising potential for the identification and detection of 14 types of GM soybean events and their derivatives. [ABSTRACT FROM AUTHOR]

BACKGROUND: Phytophthora spp. represent a pivotal genus of plant pathogens with a global distribution, exerting significant deleterious effects on food safety and forestry ecosystems. Numerous pathogenic and invasive Phytophthora species, introduced through imported fruits, have been frequently detected at Chinese ports. With the rise in global trade activities, the plant quarantine of imported fruits is becoming increasingly important but challenging. Fast, simple, and labor‐saving techniques are necessary and anticipated. RESUITS: A polymerase chain reaction restriction fragment length polymorphism capillary electrophoresis (PCR‐RFLP‐CE) technology‐based quarantine approach was developed for 16 Phytophthora species associated with the imported fruits in China. The Ypt1 gene, exhibiting abundant interspecific variations, was selected as the marker gene for PCR. The restriction endonuclease AluI was proven to be capable and compatible in simultaneously separating different Phytophthora species during CE. By combining with a fast and efficient DNA extraction kit, the developed PCR‐RFLP‐CE technique was successfully applied to identify Phytophthora species in artificially infested fruits. CONCLUSION: We provide a quick, practical, and high‐throughput detection approach for hazardous and invasive Phytophthora species associated with imported fruits in China. This strategy can give good convenience and technological support for carrying out massive quarantine activities at Chinese ports. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Ageing is a complex biological process with variations among individuals, leading to the development of ageing clocks to estimate biological age. Glycans, particularly in immunoglobulin G (IgG), have emerged as potential biomarkers of ageing, with changes in glycosylation patterns correlating with chronological age. For precision analysis, three different plasma pools were analysed over 26 days in tetraplicates, 312 samples in total. In short-term variability analysis, two cohorts were analysed: AstraZeneca MFO cohort of 26 healthy individuals (median age 20) and a cohort of 70 premenopausal Chinese women (median age 22.5) cohort monitored over 3 months. Long-term variability analysis involved two adult men aged 47 and 57, monitored for 5 and 10 years, respectively. Samples were collected every 3 months and 3 weeks, respectively. IgG N-glycan analysis followed a standardized approach by isolating IgG, its subsequent denaturation and deglycosylation followed by glycan cleanup and labelling. Capillary gel electrophoresis with laser-induced fluorescence (CGE-LIF) and ultra-performance liquid chromatography analyses were employed for glycan profiling. Statistical analysis involved normalization, batch correction, and linear mixed models to assess time effects on derived glycan traits. The intermediate precision results consistently exhibited very low coefficient of variation values across all three test samples. This consistent pattern underscores the high level of precision inherent in the CGE method for analysing the glycan clock of ageing. The AstraZeneca MFO cohort did not show any statistically significant trends, whereas the menstrual cycle cohort exhibited statistically significant trends in digalactosylated (G2), agalactosylated (G0) and fucosylation (F). These trends were attributed to the effects of the menstrual cycle. Long-term stability analysis identified enduring age-related trends in both subjects, showing a positive time effect in G0 and bisected N-acetylglucosamine, as well as a negative time effect in G2 and sialylation, aligning with earlier findings. Time effects measured for monogalactosylation, and F remained substantially lower than ones observed for other traits. The study found that IgG N-glycome analysis using CGE-LIF exhibited remarkably high intermediate precision. Moreover, the study highlights the short- and long-term stability of IgG glycome composition, coupled with a notable capacity to adapt and respond to physiological changes and environmental influences such as hormonal changes, disease, and interventions. The discoveries from this study propel personalized medicine forward by deepening our understanding of how IgG glycome relates to age-related health concerns. This study underscores the reliability of glycans as a biomarker for tracking age-related changes and individual health paths. [ABSTRACT FROM AUTHOR]

While ice has very low solubility for salts compared to water, small amounts of ions are doped into ice crystals. These small ion dopants can alter the fundamental physical and chemical properties of ice, such as its structure and electrical conductivity. Therefore, these results could have a direct impact on the chemical reactivity of ice and ice surfaces. Here, we examine the influence of the uptake of three salts—ammonium chloride (NH4Cl), sodium chloride (NaCl), and ammonium sulfate [(NH4)2SO4]—on ice Ih formation using capillary electrophoresis. Using both cation and anion modes, we observed and quantified the uptake of individual ions into the ice. Our results indicate that anions have a higher propensity for uptake into ice Ih crystals. [ABSTRACT FROM AUTHOR]

Introduction. Currently, there is an increasing trend towards assessing the full spectrum of chemical compounds in plant materials. One of the first steps in the formation of secondary metabolites of complex structure, participating in biochemical processes in the plant body, which have their own effect on the human and animal body, are organic acids (OA) and amino acids (AA). However, due attention is not always given to these compounds when studying the component composition of plant raw materials.Aim. The purpose of the work was to study the profile and quantitative assessment of the content of organic acids and amino acids of Aesculus hippocastanum L. flowers.Materials and methods. The object of the study were horse chestnut flowers, collected during flowering in the Voronezh region in 2021 and dried by air-shade method. Quantitative assessment of the amount of TC in terms of malic acid and ascorbic acid was carried out titrimetrically according to the methods presented in the State Pharmacopoeia of the Russian Federation, XIV edition. The composition of the OA and AA profiles of horse chestnut flowers was determined using the capillary electrophoresis method ("Kapel®-105/105M", Group of Companies "Lumex", Russia).Results and discussion. The method of capillary electrophoresis revealed the presence of 12 compounds of the OA group in a total amount of 4.7 % (in terms of absolutely dry raw materials). The rationality of recalculating the content of the sum of OA to the major component – citric acid, which amounted to 1.65 ± 0.04 %, was shown. The content of ascorbic acid was low (0.033 ± 0.007) %. Using the TLC method, the presence of 5 zones of compounds belonging to the AA class was established in the aqueous extract of horse chestnut flowers. The quantitative content of the sum of free AA in terms of glutamine, determined by the spectrophotometric method, was 2.25 ± 0.07 % (n = 5). The presence of 17 AA was determined by capillary electrophoresis, which amounted to 7.13 %. Glutamic acid is present in the greatest quantity in horse chestnut flowers (1.19 %). Essential amino acids amounted to 2.27 %; leucine was present in the predominant amount (0.58 %).Conclusion. A study was carried out of the qualitative composition and quantitative content of organic acids and amino acids of horse chestnut flowers. The composition of the profile and the quantitative content of organic acids and amino acids have been established. It was revealed that citric acid prevails in the total organic acids; glutamic, aspartic acid, arginine and proline are found in greater quantities in the total amino acids.

Ya-Ping Chen,1,2 Heng Wang,1,3 Lei Wang,4 Dan Xie,4 Min Guo,4 Jiang-Fen Wu,5 Bang-Quan An,6 Sheng-Wen Huang1,5 1School of Clinical Laboratory Science, Guizhou Medical University, Guiyang, Guizhou, People’s Republic of China; 2Department of Clinical Laboratory, the Second People’s Hospital of Guiyang, Guiyang, Guizhou, People’s Republic of China; 3Guiyang Public Health Clinical Center, Guiyang, Guizhou, People’s Republic of China; 4Medical College of Guizhou University, Guiyang, Guizhou, People’s Republic of China; 5Medical Genetics Department, Guizhou Provincial People’s Hospital, Guiyang, Guizhou, People’s Republic of China; 6Department of Blood Transfusion, Guizhou Provincial People’s Hospital, Guiyang, Guizhou, People’s Republic of ChinaCorrespondence: Sheng-Wen Huang, Medical Genetics Department, Guizhou Provincial People’s Hospital, No. 83, East Zhongshan Road, Nanming District, Guiyang, Guizhou, 550002, People’s Republic of China, Tel +86-851-85935128, Fax +86-851-85937194, Email hsw713@sina.comPurpose: To analyze the composition of abnormal hemoglobin and the relationship between genotype and phenotype by screening abnormal hemoglobin in a subpopulation of Guizhou, China.Patients and Methods: Routine blood evaluation, capillary electrophoresis of hemoglobin, and mutation of α - and β - thalassemia genes were evaluated in 19,976 individuals for thalassemia screening in Guizhou. Sanger sequencing of HBA1, HBA2 and HBB genes was performed in samples with abnormal bands or unexplained increases of normal bands. The types of abnormal hemoglobin were obtained by sequence analysis.Results: Abnormal hemoglobin was detected in 84 individuals (detection rate, 0.42%). Ten types each of α and β globin chain variants were detected, including most commonly Hb E, Hb New York and Hb Port Phillip. In this study, the abnormal Hb Mizuho was identified for the first time in a Chinese population, and a novel abnormal hemoglobin Hb Guiyang (HBA2: c.151C > A) was detected for the first time. Except for Hb Mizuho, other abnormal hemoglobin heterozygotes without thalassemia or iron deficiency had no significant hematological changes.Conclusion: This study enriched the molecular epidemiological data of abnormal hemoglobin in Guizhou, China and provided reference data for genetic counseling and prenatal diagnosis of abnormal hemoglobin.Keywords: abnormal hemoglobin, routine blood examination, capillary electrophoresis, DNA sequencing, Hb Mizuho, HBA2

Chronic myeloid leukemia (CML) is associated with several breakpoint regions that result in different BCR::ABL1 fusion transcripts. These include the major breakpoint region (M‐BCR), minor breakpoint region (m‐BCR), and mu breakpoint region (u‐BCR) corresponding to p210, p190, and p230 fusion transcripts, respectively. This patient is a 38‐year‐old female with a new diagnosis of CML in chronic phase. A novel p210 fusion transcript splice variant was detected with qualitative reverse transcription PCR and capillary electrophoresis. Subsequent FISH study was performed, which revealed 86.5% positive for the BCR::ABL1 fusion. Quantitative real‐time polymerase chain reaction (PCR) showed a negative result for the p210 fusion transcript. The variant was further characterized by Sanger sequencing. This variant is in‐frame and predicted to be functional. This case illustrates the need for a combination of different testing techniques to fully characterize the rare BCR::ABL1 fusion transcripts. [ABSTRACT FROM AUTHOR]

Gangliosides are sialic acid-containing glycosphingolipids that play an essential role in many biological and pathophysiological processes. They are present in high amounts in the central nervous system and their abnormal metabolism or expression has been observed in many diseases. We have developed and validated a sensitive capillary electrophoresis laser-induced fluorescence (CE-LIF) method for the separation and quantification of oligosaccharides digested from nine gangliosides of high biological relevance. APTS was used for the labeling of the glycans. Reverse polarity CE was performed for the separation of the labeled glycans bearing negative charges. The optimized background electrolyte is a 15 mM lithium acetate buffer with pH of 5 containing 5% w/v linear polyacrylamide, which allows for the separation of all nine gangliosides. Validation parameters including linearity, precision, and accuracy were evaluated. LOQ and LOD were in the nM range, comparable to those of LC-MS techniques. The method was used to identify and quantify the ganglioside pattern of glioblastoma and neuroblastoma cell lines. The presented method is a valuable tool for further investigations aiming at understanding the role of gangliosides in various neurological diseases or CNS tumors. [ABSTRACT FROM AUTHOR]

Hemoglobin Shaare Zedek (Hb SZ) is a rare structural α-Hb variant. Characterizing its genotype–phenotype relationship and genetic origin enhances diagnostic and clinical management insights. We studied a proband and six family members using high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), PCR, and sequencing to analyze α- and β-globin genes and α-globin haplotypes. Pathogenicity predictions and a rapid diagnostic method were developed. The proband, his father, grandfather, and aunt had Hb migrating to the HbH-zone on CE and elevated fetal hemoglobin (HbF) on HPLC. Direct sequencing identified an A to G mutation at codon 56 of the α2-globin gene, characteristic of Hb SZ. Additionally, the proband carried a β-globin gene mutation [HBB.52A>T]. Mild thalassemia-like changes were observed in the proband, whereas individuals with only the Hb SZ variant did not exhibit these changes. Pathogenicity predictions indicated that Hb SZ is benign. The variant can be identified using restriction fragment length polymorphism (RFLP) and allele-specific PCR. The Thai variant of Hb SZ is associated with the haplotype [- - M - - - -]. Hb SZ is a non-pathological variant that minimally affects red blood cell parameters, even when it coexists with β0-thalassemia. HPLC and CE systems cannot distinguish it from other Hbs, necessitating DNA analysis for accurate diagnosis. [ABSTRACT FROM AUTHOR]