Acrylamide is a mutagenic and likely carcinogenic molecule. Incomplete polymerization of commercial acrylamide-based polymers leaves residual acrylamide molecules that pose a threat to human health and the environment. Acrylamide, being a polar molecule, is readily soluble in water but cannot be easily ionized in an aqueous environment. Therefore, the conventional capillary zone electrophoresis method is challenging to apply to acrylamide molecules. In this study, a capillary electrophoresis method that alters the acid-base properties of acrylamide under nonaqueous conditions, thereby imparting electrophoretic mobility to the molecule and increasing sensitivity through sample stacking, was used to detect acrylamide residues in absorbent polymers used in baby diapers. The method exhibited excellent analytical performance, with a linear range of 20-500 µg/L (R[2] = 0.9999), limits of detection and quantification of 5.3 and 17.7 µg/L, respectively, and satisfactory precision and recovery values. The applicability of the proposed approach was demonstrated by analyzing five commercial baby diaper samples. Acrylamide was detected in all samples at concentrations ranging from 0.483 to 1.36 µg/g, indicating the presence of residual monomer associated with polyacrylamide-based superabsorbent polymers. These findings highlight the capability of combining nonaqueous capillary electrophoresis (NACE) with online preconcentration techniques for the sensitive determination of acrylamide in complex polymer-based matrices and emphasize the importance of monitoring such residues in hygiene products intended for infant use.

The environmental persistence of microplastic particles has raised increasing concerns for both human health and ecological systems. Owing to their intrinsic electrical properties and broad size distribution, electrokinetic-based microfluidic separation techniques have attracted increasing attention. In this study, a hybrid dielectrophoresis-bipolar electrode (DEP-BPE) microchannel is proposed to enhance particle separation across multiple size scales. A multiphysics numerical model coupled with particle tracking is developed to investigate the coupled electrokinetic mechanisms governing particle transport, including dielectrophoretic (DEP) force, electrophoretic migration, and electroosmotic flow (EOF)-induced hydrodynamic drag. The results show that increasing the applied voltage amplifies electrokinetic forces and significantly improves removal efficiency. However, the separation performance exhibits a non-monotonic dependence on voltage, arising from the competition between lateral DEP displacement and axial transport driven by electrophoresis and EOF. An optimal voltage condition is identified, under which trajectory differentiation is maximized without over deflection. Electrode spacing is found to primarily regulate the spatial distribution of electrokinetic interactions, exerting a limited influence on overall separation strength but significantly affecting the location of particle deflection and trajectory evolution. These findings demonstrate that particle separation in the DEP-BPE microchannel is governed by the coupling between force magnitude and spatial distribution and highlight the effectiveness of integrating DEP-induced stratification with BPE-driven trajectory divergence. This work provides mechanistic insights and design guidelines for high-efficiency microfluidic separation of polydisperse microplastic particles.

Vitamin D3 (VD3) and its metabolite 25-hydroxyvitamin D3 (25OHD3) are important biological molecules that play a significant role in human well-being. Conventional methods use highly hazardous solvents to extract them from their bound form, which is prior to their analysis in high-performance liquid chromatography (HPLC). Considering the World Health Organization's (WHO's) sustainable development goals (SDGs), there is a need for green solvents to extract valuable biological metabolites such as VD3 from human plasma. This study explored the potential of existing green solvents to liberate VD3 and 25OHD3 in human plasma samples. Five green solvents were shortlisted based on their physicochemical properties and low toxicity level. HPLC-UV chromatography conditions using a C8 column (BEH C8 1.7 µm, 2.1 × 150 mm[2]) were identified to achieve good peak-to-peak separation of 25OHD3 (2.74 min) and VD3 (4.5 min) within 5 min. The optimized method had an excellent limit of detection (LOD) for both VD3 (86 pg/mL) and 25OHD3 (89 pg/mL). Ethyl acetate has outperformed other green solvents and a solvent mixture of acetonitrile and methanol (in different ratios) in extracting both VD3 and 25(OH)D3 from human plasma samples (RSD < 2.2%). The stability of VD3 and 25OHD3 was tested with different plasma storage conditions (5°C, -20°C, -80°C, and lyophilized powder) for up to 60 days. Plasma stored at -20°C has significantly preserved the stability of VD3 and its metabolite 25OHD3, both of which were successfully extracted using ethyl acetate with minimal loss even on the 60[th] day of storage.

This study presents an affordable micellar electrokinetic chromatography (MEKC) method for the simultaneous analysis of six commonly used antiepileptic drugs: ethosuximide, phenobarbital, lamotrigine, phenytoin, carbamazepine, and diazepam. Systematic optimization of the method resulted in a running buffer containing 80 mM sodium dodecyl sulfate, 5% methanol, 2.5 mM phosphate, and 10 mM borate at pH 8.7, which provided robust and reliable separation. The method demonstrated excellent precision, with migration time %RSD values below 2% for both intraday and inter-day repeatability, and peak area %RSD values within acceptable limits (1.7%-3.7% intraday; 2.9%-6.3% inter-day). Linearity was confirmed for all analytes, with r[2] values exceeding 0.99 and residuals randomly distributed around zero. Robustness testing showed that small variations in electrophoretic conditions did not significantly affect the resolution of the critical analyte pairs, which consistently remained above 1.5. Application of the method to five antiepileptic tablet formulations obtained from the Ethiopian market demonstrated its suitability as a rapid, reliable, and cost-effective screening tool. Overall, the developed MEKC method provides a practical analytical approach for antiepileptic medicines' quality assessment in resource-limited settings, supporting efforts to strengthen regulatory surveillance and safeguard public health.

The potential of multi-capillary electrophoresis (CE) for high-throughput analysis of legacy CE applications was investigated by optimisation of the separation of six pharmaceutical compounds on a single-capillary instrument, followed by transference of the method to an eight-capillary BioPhase 8800 system from SCIEX. Multi-CE had superior performance, allowing construction of calibration curves in a single run, and a five-fold reduction of run time per sample, improved linearity, accuracy and limits of detection. Method transfer from the single-capillary system required minor instrumental parameter adjustments using the same background electrolyte. The potential of multi-CE was shown via quantification of caffeine in a variety of energy drinks, demonstrating broad applicability where high-throughput and rapid analyses are desirable.

Gallium-based liquid metals exhibit attractive properties for precision manipulation, combining high fluidity, superior electrical conductivity, and electrically tunable interfacial behavior. Their deformable liquid interfaces provide a promising route for adaptive contact with irregular, fragile, or soft objects. Conventional rigid fixtures may induce surface microscratches, structural damage, or residual contamination when manipulating precision components, such as glass microspheres or optical elements, limiting their use in high-precision assembly. To address these limitations, we propose a compact flexible gripper system that uses an electrically driven Galinstan droplet as the primary actuating element. By regulating the interface between the droplet and the electrolyte through coupled electrocapillary and electrochemical processes, the liquid metal undergoes controllable deformation, enabling stable gripping, holding, and release. Under reverse voltage, the interfacial state of the droplet recovers, allowing the droplet to retract and release the object without visible residue. Integrated with a compact motion platform, the system enables automated transport of small objects under controlled experimental conditions. The effects of applied voltage, NaOH concentration, electrolyte volume, electrode size, clamp geometry, and Galinstan droplet volume were systematically investigated to evaluate the gripping and release performance. The gripper achieved stable clamping within 0.2-4 s and exhibited a relatively long stable holding duration at a low voltage of approximately 0.15 V. In addition to precision glass microspheres, the gripper successfully transported representative objects with different material properties and geometries, including ceramic spheres, iron spheres, and small fish specimens with characteristic sizes of approximately 2 mm. Load-bearing tests further identified a maximum stable transportable load of 0.4 g for the gripper with a 2.25 mm aperture. This work provides a feasible strategy for electrically controlled and gentle manipulation of small objects, demonstrating the potential of Galinstan droplet grippers in precision assembly and small-scale automated handling.

Accurate particle characterization is critical for biomedical research, yet electrical impedance flow cytometry suffers from measurement inaccuracies due to the positional heterogeneity of particles within microfluidic channels. In large-scale microchannels, off-axis particle trajectories lead to significant waveform distortion and the attenuation of key characteristic features, undermining measurement precision. To address this issue, this study presents a novel signal-processing framework centered on a full-waveform calibration factor (CF). The CF is theoretically grounded in the exponential decay of the electric field along the vertical axis of coplanar electrodes and is derived from comprehensive waveform features rather than localized extrema, ensuring robustness against the distortions prevalent in high-flow conditions. Coupled with a dedicated real-time algorithm, it corrects for vertical position-induced errors in particle sizing. The effectiveness of this method was validated through a combination of finite element simulation and experimental impedance measurement. The system's performance is demonstrated by accurately discriminating polystyrene microsphere populations (10 μ m , 15 μ m , and their mixture) and by resolving the size distributions of viable versus non-viable HEK-293T cells. Statistical analysis (N ≥ 500 events per group, p < 0.001) confirms significant improvement in size-distribution accuracy after calibration. Furthermore, this study designs an inverted microfluidic chip enabling real-time, optical sensing of particle stream height within the impedance detection zone and supporting position-independent impedance cytometry applicable to continuous-flow particle streams. Experimental calibration achieves a strong linear correlation (R 2 = 0.970) between the sensor response and measured height, confirming the method's utility for spatially resolved impedance cytometry.

In this study, a capillary electrophoresis (CE) method was developed for the determination of naltrexone, an opioid antagonist, and its primary metabolite, 6-β-naltrexol (6-β-NTX). Naltrexone is commonly used in the treatment of opioid and alcohol dependence. Accurate quantification of such drugs and their metabolites is essential for ensuring therapeutic efficacy and safety. Reliable determination of NTX and 6-β-NTX in plasma is important for bioanalytical and clinical investigations involving naltrexone therapy. Method optimization was carried out using Box-Behnken design within the framework of response surface methodology to achieve maximum resolution between naltrexone and 6-β-naltrexol. Separation was performed at 15°C using a 10 mM phosphate buffer (pH 2.0) under an applied voltage of 20 kV, without the addition of organic modifiers. The method was validated in accordance with bioanalytical validation guidelines, providing a validated lower limit of quantification (LLOQ) of 0.05 µg/mL for both analytes, with precision (% relative standard deviation [RSD]) below 7% at the LLOQ. The validated method was successfully applied to plasma samples obtained from patients receiving treatment for alcohol and opioid use disorders. Furthermore, the environmental sustainability of the proposed CE method and its sample preparation steps was assessed using green analytical chemistry metrics, such as AGREE, AGREEprep, and MoGAPI, demonstrating its greener profile compared to reported methods.

Globally, cancer morbidity and death are rising quickly, making it imperative to develop a workable plan for early cancer patient identification and treatment outcome prediction. Liquid biopsy, a minimally invasive and repeatable technique, can replace the restrictions of tissue biopsy by detecting, assessing, and tracking cancer in any bodily fluid, including human blood. This novel diagnostic technique offers crucial data for tracking and recognizing tumor genomes in samples of physiological fluids. Through minimally invasive methods, the identification of tumor-origin biological components, such as circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), circulating microRNAs (miRNAs), exosomes, and autoantibodies in serum, plasma, saliva, and urine, allows for disease monitoring, early diagnosis, and assessment of treatment response. The goal of this study is to provide a comprehensive summary of the most recent discoveries and applications of liquid biopsy technology for detecting cancer biomarkers, with an emphasis on the years 2005-2025. When used in conjunction with other test results, liquid biopsy is also advantageous for early recurrence detection, monitoring patients who are considered to be at high risk of getting cancer and improving diagnosis. Combining patient history, clinical results, and biopsy biomarker amounts will enable more precise and timely diagnosis thanks to developments in analytical methods and informatics. This review thoroughly examined the biosynthesis, significance, and potential role of four well-known biological markers in tumor diagnosis and treatment. In addition, the advances and challenges in detection technologies for these analytes, as well as potential solutions to address these issues, were also discussed. Finally, we discuss the advantages and disadvantages of liquid biopsy, along with suggestions for its future advancement.

We developed an imaging-based approach to directly visualize and monitor molecular migration during capillary electrophoresis and applied it to the separation of monoclonal antibodies (mAbs) by capillary gel electrophoresis with sodium dodecyl sulfate (CGE-SDS). Proteins were detected through their intrinsic fluorescence at 350 nm following excitation at 280 nm, enabling real-time observation of migration behavior without additional labeling. Time-resolved imaging allowed direct visualization of both migration and band broadening of mAbs under reducing and non-reducing conditions throughout the separation process. Analysis of electrophoretic mobilities suggests that mAb separation in the gel matrix is largely governed by the Ogston sieving mechanism. Quantification of band broadening revealed that peak dispersion increased with applied electric field strength, as evidenced by the marked reduction in broadening when the separation voltage was removed. Qualitative evaluation of the dispersion behavior suggests that mAb band broadening arises from the combined effects of electromigration dispersion (EMD) and gel-sieving-induced dispersion, with glycosylation-related heterogeneity providing an additional contribution for glycosylated species such as the heavy chain (HC) and intact mAb. These results demonstrate the utility of real-time imaging for elucidating separation mechanisms and dispersion phenomena in CGE-SDS and provide new insights into factors governing the electrophoretic behavior of therapeutic antibodies.

Thyroglobulin (Tg) is a clinically established biomarker for thyroid cancer; however, its diagnostic specificity remains limited due to confounding benign conditions. This study investigated changes in the site-specific N-glycosylation profiles of thyroglobulin protein derived from papillary thyroid cancer and adjacent healthy tissues. Using a bottom-up mass spectrometry-based glycoproteomic approach, we analyzed site-specific glycan compositions in retrospective patient tissue samples. Nine distinct N-glycosylation sites were identified and quantified in this study. Statistical analysis revealed significant differences in the site-specific glycosylation profiles. The glycan compositions HexNAc(3)Hex(5) and HexNAc(3)Hex(6) at the Asn1365 site exhibited a discriminatory potential. Dimensionality reduction and hierarchical clustering based on the top four glycopeptide variants showed observable grouping of tumor versus normal samples. Furthermore, supervised machine learning models were used to evaluate these features, with a neural network achieving the highest diagnostic performance (area under the curve [AUC] = 0.83). Feature importance analysis indicated that Asn1365-HexNAc(3)Hex(6) was the strongest contributor to the predictive power of the model. The observed alterations in specific glycan compositions, combined with machine learning evaluations, may offer improved diagnostic specificity and provide promising avenues for personalized thyroid cancer management.

Free-flow electrophoresis is a core technology for biological particle separation, but traditional devices suffer from low automation, poor flow stability, and limited throughput, hindering industrial-scale applications. Herein, a novel fully automatic reciprocating free-flow isoelectric focusing (RFFIEF) system, a batch-type of automated closed-loop free-flow isoelectric focusing (FFIEF), was developed, integrating a 24-channel separation chamber, closed gas-liquid circulation drive, and intelligent control system to achieve unattended standardized operation. Standard protein verification showed excellent flow stability and pH gradient reproducibility (pooled standard deviation [SD] = 0.19 pH units excluding ampholyte-induced outliers). The system was applied to large-scale separation of milk exosomes, which were effectively focused at pH 5.0-7.5. Exosome subpopulations were fractionated by surface charge: CD81-positive subpopulations concentrated at pH 5.8-6.5, whereas CD63-positive subpopulations were highly enriched at pH 6.9. This system breaks through the automation bottleneck of conventional devices, providing an efficient platform for large-scale biological particle separation with broad industrial prospects.

Phosphonothrixin (PPX), a phosphonate natural product known for its potent herbicidal activity, has attracted renewed interest because of the need for alternative herbicides. Its biological activity is confined to the (S)-enantiomer. Although the only reported enantioselective synthesis is complex, labor-intensive, and low yielding, direct chiral liquid chromatographic separation is likewise challenging because of its high polarity and lack of aromatic moieties, which limit interactions with commonly used chiral stationary phases (CSPs). In addition, phosphonic acid-containing compounds tend to chelate metal ions and adsorb to metal surfaces, further complicating chromatographic separation by causing pronounced peak tailing. Here, we describe a reliable and reproducible analytical and preparative method for the direct chiral separation of PPX using a zwitterionic sulfohexyl quinidine and quinine carbamate CSPs, as well as two synthetic phosphonate derivatives using a cellulose tris(3,5-dichlorophenylcarbamate) stationary phase as an alternative approach. The best separation of PPX was achieved with the zwitterionic sulfohexyl quinidine CSP (Rs = 2.4). The separated enantiomers were isolated, and their chemical and stereochemical identities were confirmed by mass spectrometry, optical rotation, and electronic circular dichroism spectroscopy. This approach enabled the successful isolation of enantiomerically pure (S)-(-)- and (R)-(+)-PPX.

This study evaluated kinship identification using 19 short tandem repeats (STRs) from AGCU EX22 kit, 119 microhaplotypes (52 in Panel A + 67 in Panel B), and a theoretical 9622 genome-wide single nucleotide polymorphisms (SNPs) panel (The 9K). The system power was assessed accounting for linkage on the basis of 198 Chinese Southern Han (CHS) family samples and simulated data. STR + Panels A and B distinguished full-sibling (FS) from unrelated (un), with second-degree effectiveness above 0.96 when log10(LR) threshold was 4 and low false positive rate (<0.06%), but were insufficient for more distant kinship identification alone. The 9K panel was sufficient to separate third-degree from un but not for fourth-degree or more distant relatives. For discrimination among different kinships, STR + Panels A and B successfully distinguished all FS from half-siblings (HS), uncle/aunt-nephew/niece (UN), and grandparents-grandson/granddaughter (GS) in real data, with minimal overlaps (<0.2%). Discriminating second- and third-degree or within second-degree relatives seems very challenging, even for 9K panel. Including a carefully selected additional relative (AR) may improve discrimination between GS and UN/HS, such as AR (GS-un, which is GS for one person and un for the other) and AR (parent/child (PC)-UN). Similarly, an AR (PC-GS) may help distinguish UN from GS/HS, but there was slightly improved efficiency for HS.

Electrokinetic extraction (EKE), coupled with polymer inclusion membranes (PIMs) for colourimetric detection, offers a promising approach for extracting metal ions from solid matrices, such as sand, without sample pretreatment. Our EKE system consists of a formic acid gel positioned at the cathode side to neutralise hydroxide ions generated by electrolysis during the EKE of Cu(II), whereas the anode side contains the sample compartment, prepared from a syringe tube into which the sample is placed. The PIM is sandwiched between these compartments to accumulate Cu(II) via EKE through the sand and into the PIM within which it complexes with a colour-forming reagent, 1-(2-pyridylazo)-2-naphthol (PAN), enabling quantification via colourimetric analysis. The system has been optimised to expand the quantification range of Cu(II), thus allowing greater applicability to a wider range of samples with different levels of heavy metals. Increasing the column diameter extended the upper limit of the quantification range from 7.5 mg kg[-1] (4.5 mm column diameter) to 30.0 mg kg[-1] (13.5 mm column diameter). Concurrently, optimisation of the PIM composition, containing 43.3 wt% cellulose triacetate (CTA)-56.0 wt% di-(2-ethylhexyl) phosphoric acid (D2EHPA)-0.8 wt% PAN, enhanced the overall recovery of Cu(II) from 64% ± 7% to 90% ± 4%. Overall, the optimised system has achieved a quantification range of 2.8-30.0 mg kg[-1] of Cu(II), with an LOD of 0.9 and LOQ of 2.8 mg kg[-1] of Cu(II).