Analytical Methods最新文献

Correction for ‘A portable fluorescent aptamer sensor for rapid quantitative detection of Hg²⁺’ by Jiayi Li et al., Anal. Methods, 2025, 17, 4461–4469, https://doi.org/10.1039/D5AY00115C.

In this research, we developed a rapid assay for detecting human adenovirus (HAdV) and its prevalent subtypes (including HAdV3, HAdV4, HAdV7, HAdV14, HAdV11, and HAdV55) based on recombinase-mediated isothermal amplification combined with lateral flow chromatography (RAA-LFD). We designed specific primers and probes targeting conserved regions of the Hexon genes of HAdV and its common subtypes, and established the recombinase-aided amplification (RAA) reaction system. The optimal primer and probe combinations were identified utilizing real-time fluorescence RAA. Subsequently, the RAA-lateral flow dipstick (RAA-LFD) reaction system was developed and optimized for reaction temperature. At the same time, the sensitivity and specificity of the method were evaluated, and 50 clinical samples were analyzed. The optimal amplification temperature for the RAA-LFD assay was determined to be 37 °C, with a detection limit of 10–100 copies per µL, high specificity, and no cross-reactivity with other pathogens. Following processing of 50 clinical respiratory specimens using our laboratory's proprietary trehalose ester-based extraction-free reagent, the RAA-LFD assay was performed and compared with conventional qPCR, showing complete agreement between the two methods (kappa value of 1). In conclusion, this study established a simple, rapid, visual, sensitive, and specific method for the detection of HAdV and its common subtypes.

Illegal adulteration of traditional Chinese medicines has become a critical global safety concern, driven by the covert addition of synthetic pharmaceuticals such as phosphodiesterase type five inhibitors, anorectic agents, nonsteroidal anti-inflammatory drugs, corticosteroids, and sedatives. This review critically examines the rapid expansion of ambient ionization mass spectrometry technologies and evaluates their performance in detecting diverse adulterants across powders, pills, decoctions, creams, and botanical tissues. Techniques including desorption electrospray ionization, direct analysis in real time, wooden tip electrospray ionization, paper spray ionization, thermal desorption electrospray ionization, low temperature plasma ionization, and dielectric barrier discharge ionization demonstrate high sensitivity, structural specificity through tandem mass spectrometry, and near zero sample preparation, enabling analysis within seconds. Evidence from the past decade shows strong concordance between ambient ionization mass spectrometry screening and laboratory based chromatographic confirmation, highlighting its transformative role in high throughput surveillance, border inspection, and emergency toxicology diagnostics. The review further analyzes limitations related to matrix effects, quantitative variability, identification of novel analogues, and challenges in regulatory acceptance. Overall, ambient ionization mass spectrometry represents a significant advance for rapid front line detection of pharmaceutically adulterated herbal products, offering a scalable and versatile platform that strengthens public health protection.

Staphylococcal enterotoxin B (SEB) is a major pathogenic factor in foodborne diseases, and its rapid and sensitive detection is crucial for early warning of food safety risks and public health protection. In this study, a colorimetric immunosensing system for SEB detection was constructed based on a magnetic covalent organic framework (COF) composite loaded with platinum nanoparticles (Pt NPs) that exhibit dual enzymatic activities. The effective loading of Pt NPs endows the composite with excellent peroxidase (POD) and oxidase (OXD)-like activities. The COF matrix not only provides abundant binding sites for Pt NP dispersion but also enhances substrate adsorption, thereby significantly improving catalytic efficiency. By introducing an Fe₃O₄ core, the nanocomposite enables facile preparation of antibody–nanoparticle conjugates through separation and enrichment functions. The Fe₃O₄@COF@Pt nanocomposite demonstrates excellent peroxidase- and oxidase-like activities, capable of decomposing H₂O₂ and O₂ to generate hydroxyl radicals (˙OH) and superoxide anions (O₂˙⁻), which catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxidized TMB (_OXTMB), resulting in a visible color change for signal output. The sensor achieves specific detection by utilizing its peroxidase-mimicking activity, exhibiting a linear detection range of 1–200 ng mL⁻¹ with a detection limit as low as 0.395 ng mL⁻¹. This nanozyme-linked immunosorbent assay (NLISA) based on Fe₃O₄@COF@Pt offers a highly sensitive and convenient platform for SEB detection.

Glycosylated RNAs (glycoRNAs) are a groundbreaking discovery in glycobiology, extending the diversity of known glycoconjugates beyond proteins and lipids. Accumulating evidence indicates their pivotal roles in cellular communication, immune regulation, and disease pathogenesis, highlighting their promise as novel biomarkers and therapeutic targets. Robust and sensitive detection methods are crucial for deciphering glycoRNA glycosylation patterns and elucidating their physiological and pathological functions. Despite the development of diverse analytical techniques, a systematic evaluation of glycoRNA detection strategies remains lacking. To address this gap, in this review, we comprehensively summarize recent methodological advances, categorizing the published approaches based on their glycan-targeting recognition mechanisms and critically assessing their principles, applications, strengths, and limitations. We also sketch the key challenges and future development directions. This review aims to provide a timely and informative guide for researchers in this rapidly evolving area.

A rapid method has been developed for the determination of 17α-ethinylestradiol and drospirenone in contraceptive formulations. The method is based on the direct coupling of a programmed temperature vaporizer inlet to a quadrupole mass spectrometer via a deactivated fused silica tube (10 m × 0.18 mm) that is maintained at 275 °C throughout the entire analysis. The inlet is equipped with a baffled glass liner coated with Siltek™ and the injection is performed in split mode (ratio 1 : 10). The data acquisition time was only 1.0 min per sample, allowing for the high-throughput quantification of active principles in contraceptive pills in minimal time. The goal is to apply this strategy as a screening tool in quality control processes, limiting the use of the more costly and time-consuming chromatographic methods to those pills showing anomalous results, thus optimizing laboratory resources. Sample treatment involved a fast ultrasound-assisted solid–liquid extraction, and both sample preparation and instrumental conditions were optimized. The determination of the active principles was performed using the standard addition method. Accuracy, expressed as recovery percentage relative to the mass of active principle declared by the manufacturer, varied between 80% and 120%. Intra- and inter-day precision were adequate, with values of relative standard deviation (% RSD) equal to or less than 9% and 11%, respectively. Furthermore, the sustainability profile of the proposed rapid method was evaluated using the HEXAGON algorithm, which demonstrated that the PTV-MS method contributes to sustainable development by benefiting both the environment and society.

Camellia oil is highly valued for its nutritional benefits, but its premium market position has led to frequent mislabeling of geographical origin. To assess the feasibility of tracing camellia oil origin using GC-IMS combined with chemometrics, 66 samples from 11 representative production areas across 7 provinces in China were collected and analyzed. Clustering and discriminant analyses were conducted at both the province and city levels. PCA revealed clear clustering by province, with even greater differentiation when the origin was specified at the city level. The PLS-DA model based on VOC profiles achieved recognition accuracies of 96.7% (province level) and 100% (city level), while the successive projections algorithm (SPA)-PLS-DA model achieved 100% and 98.5%, respectively. The findings demonstrate that GC-IMS combined with chemometrics is a feasible and effective method for camellia oil origin traceability, providing technical support for the certification of geographical indications and food safety supervision.

Adsorbents with significant adsorption capacity and high selectivity are required to identify and eliminate heavy metal ions from the environmental and food samples under study. For this purpose, in the present work a new adsorbent, graphitic carbon nitride g-C₃N₄ hybrid with CaAl layered double hydroxide, was effectively synthesised using an easy hydrothermal co-precipitation procedure and applied for Pb²⁺ determination in food and water samples using dispersive solid phase-microextraction (DSP-µE) via FAAS. The as synthesized CaAl-LDH@g-C₃N₄ nanocomposite characteristics were examined using XRD, SEM, SEM-EDX and FTIR. Significant analytical parameters were optimised such as pH 8, adsorbent quantity 10 mg, and adsorption and elution times 30 and 60 s, respectively. Additionally, a sample volume of 20 mL and an eluent volume of 2 mL were used with an elution solvent of 0.5 M HNO₃. A matrix effect study was also performed as verification of the developed method. A limit of detection (LOD) of 0.54 µg L⁻¹, a limit of quantification (LOQ) of 1.79 µg L⁻¹, an enrichment factor (EF) of 10.69 and a preconcentration factor (PF) of 10 were determined. The % RSD (< 10%) with an R² value of 0.999 has proven the linearity and high accuracy of the developed method. ComplexMoGAPI evaluation (score 79) confirmed the excellent greenness of the proposed method. Standard reference materials (SRMs) were used to affirm the accuracy of the method. Different instant noodle samples, and tap water samples were analysed for their Pb²⁺ content, allowing for the successful determination of Pb²⁺ levels in them.

Dipeptidyl peptidase-IV (DPP-IV) is critical for drug metabolism and physiological regulation. In terms of differences in DPP-IV among species, mouse and human DPP-IV share a high degree of similarity, and mice are typically used as the optimal animal model for conducting the relevant pharmacological evaluations and biological function studies. Herein, a detection method is developed using the fluorescent probe GP-BAN and mouse tissue S9 fraction (enzyme source) to rapidly quantify DPP-IV activity in mouse tissues. The method was validated for specificity, linearity and precision, and the metabolite BAN showed good linearity in the 0–20 µM range using a weighted (1/x) least squares linear regression model (r² = 0.9996), with an LOD of 5.5 nM and LOQ of 16.7 nM. In C57 mice, the DPP-IV activity in 14 tissues/organs (including the liver, kidney, thymus, and small intestine) differed significantly: the thymus had the highest activity (2.64 nM µg⁻¹ protein per min), followed by the liver, kidney and small intestine. Enzyme kinetics showed that the K_m of GP-BAN for mouse DPP-IV was 34.05 µM, which is close to that of human liver microsomes (HLM, 41.46 µM), indicating cross-species substrate binding consistency. ELISA confirmed that DPP-IV protein expression correlated positively with activity in the liver, kidney and thymus (r > 0.92, p < 0.001). This sensitive, species-translatable assay and its organ-specific activity/kinetic data support mouse models in preclinical DPP-IV studies, improving cross-species extrapolation predictability in physiological research.

This paper describes a new analytical approach for determining gamma-hydroxybutyric acid (GHB) in biosamples using liquid chromatography-mass spectrometry (LC-MS) following derivatization with N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDC). The derivatization procedure is simple, rapid, reproducible, inexpensive, and safe. It allows for a complete variation of GHB chemical characteristics (from hydroxyacid to N-acylurea) with a molecular weight increase from 104 to 259 u and the introduction of 3 nitrogen atoms. These modifications promote the protonation of the analytes in the MS electrospray ion source, enabling MS detection under positive ionization conditions. Liquid chromatography-high-resolution accurate-mass Orbitrap mass spectrometry (LC-HRAM-Orbitrap-MS) measurements demonstrated that derivatization produces two GHB-EDC derivatives with the same exact mass (MH⁺ ions at m/z 260.1968) and experimental isotopic patterns overlapping each other and superimposable to the calculated one. Thus, they share an identical elemental composition (C₁₂H₂₅O₃N₃) but have different molecular structures (GHB-EDC_A and GHB-EDC_B). Equivalent results were obtained for D₆-GHB: the production of two deuterated N-acylureas (D₆-GHB-EDC_A and D₆-GHB-EDC_B), with MH⁺ ions at m/z 266.2341, superimposable experimental and calculated isotopic patterns, elemental composition C₁₂H₁₉D₆O₃N₃, and different molecular structures, mirroring those of GHB-EDC_A and GHB-EDC_B. After optimizing the derivatization conditions (reaction solvent, reaction temperature and time, and volume and concentration of the derivatizing agent) the final procedure involves reacting with 10 mM aqueous EDC, at 45 °C for 15 minutes. GHB-EDC derivatives were found to be highly stable over time (at least 15 days), even at room temperature. Three preliminary analytical methods for the determination of endogenous and exogenous GHB levels in urine, blood, and hair samples were developed.