Detection Limit Research Articles

Programming the Dynamic Range of Nanochannel Biosensors for MicroRNA Detection Through Allosteric DNA Probes

AbstractSolid‐state nanochannel biosensors are extensively utilized for microRNA (miRNA) detection owing to their high sensitivity and rapid response. However, conventional nanochannel biosensors face limitations in their fixed dynamic range, restricting their versatility and efficacy. Herein, we introduce tunable triblock DNA probes with varying affinities for target miRNA to engineer solid‐state nanochannel biosensors capable of customizable dynamic range adjustment. The triblock DNA architecture comprises a poly‐adenine (polyA) block for adjustable surface density anchoring, alongside stem and loop blocks for modulating structural stability. Through systematic manipulation of these blocks, we demonstrate the ability to achieve diverse target binding affinities and detection limits, achieving an initial 81‐fold dynamic range. By combining probes with various affinities, we extend this dynamic range significantly to 10,900‐fold. Furthermore, by implementing a sequestration mechanism, the effective dynamic range of the nanochannel biosensor is narrowed to only a 3‐fold span of target concentrations. The customizable dynamic range of these advanced nanochannel biosensors makes them highly promising for a broad spectrum of biomedical and clinical applications.

Analytical Assessment of the Quality of Dietary Supplements and Cosmetic Products Containing Xanthohumol by Thin-Layer Chromatography Along with the Estimation of Its Antioxidant Potential

Xanthohumol, a prenylated chalcone in the flavonoid group, naturally occurs in many plants and exhibits antioxidant, anticancer, anti-inflammatory, antibacterial, and antiviral effects. The growing interest in xanthohumol due to its potential therapeutic properties has led to the increase in the pool of products available on the market. The novelty of this study is the proposal of a rapid and cost-effective procedure useful for performing quality control on products containing xanthohumol in the form of dietary supplements and cosmetics as well as testing their stability. For this purpose, the thin-layer chromatography method with densitometric detection was used, which was validated in accordance with ICH (International Conference on Harmonization) guidelines. The mobile phase was toluene, 1,4-dioxane, and glacial acetic acid (37:10:1.5 v/v/v), and TLC silica gel 60 F254 plates were used as the stationary phase. The validation process assessed linearity, with a correlation coefficient (r) of 0.9987. The calculated LOD (limit of detection) and LOQ (limit of quantification) values were 3.82 and 11.57 ng/spot, respectively. Accuracy was evaluated by determining percentage recovery at three concentration levels (80, 100, and 120%), with an average recovery of 100% and RSD below 1%, confirming good accuracy. Precision was indicated by an RSD of less than 2.20%. The average content of xanthohumol in dietary supplements ranged from about 8 to 29% of the content declared by the manufacturers. The stability tests showed that XN decomposes most slowly in water (t0.5 = 10.86 h) compared with acidic (t0.5 = 10.80 h) and alkaline solutions (t0.5 = 7.39 h), as well as in the presence of an oxidizing agent (t0.5 = 18.38 h), at all tested temperatures, which is confirmed by the calculated kinetic parameters. In the tests of antioxidant capacity, xanthohumol shows significantly higher radical scavenging capacity than vitamin C in the entire range of analyzed concentrations (0.03–2.40 mmol/L).

Enhanced industrial wastewater monitoring: method development for non-target screening of highly polar substances using ZIC-HILIC-HRMS.

Non-target screening (NTS) plays a major role in the monitoring and management of water bodies. While the NTS of moderate to non-polar substances is well-established, the screening of highly polar chemicals remains challenging. In this study, a robust separation method for highly polar substances using zwitterionic hydrophilic interaction liquid chromatography coupled with high-resolution mass spectrometry (ZIC-HILIC-HRMS) was developed. This method was specifically designed for the NTS of industrial wastewater, with the objective of capturing a wide range of polar contaminants in each acquisition run. Method validation included assessing key parameters such as repeatability, reproducibility, linearity, and limit of detection (LOD). For repeatability and reproducibility, the average %RSD of intensity and retention time across all substances in different matrices-solvent, influent, and effluent-remained below 6% and 1%, respectively (n = 10). The method demonstrated good linearity (R2 > 0.99) for 75% of the substances, while LODs varied between 0.1 and 40µg/L depending on the compound tested. The method was then applied for NTS analysis of untreated wastewater at various locations within a chemical industrial park. Additionally, the overall influent and effluent of an industrial wastewater treatment plant (WWTP) were monitored over a 10-day period. Principal component analysis (PCA) was performed to interpret the data, identifying irregularities in the wastewater content. Moreover, the method demonstrated the WWTP's ability to achieve an average removal efficiency of approximately 90% for this category of substances in this period, while also detecting their degradation products in the effluent. Finally, the method was successfully integrated into the daily monitoring routine of the WWTP, ensuring continuous surveillance and improved management of wastewater treatment processes.

Pyroelectrically Driven Charge Transfer and its Advantages on SERS and Self‐Cleaning Property

AbstractThe fabrication of reusable SERS substrates with sensitivity at the single‐molecule level remains challenging but promising. Herein, a composite surface‐enhanced Raman scattering (SERS) substrate is proposed that includes Ag nanoparticle (Ag NPs) /Graphene/BaTiO3 (Ag/G/BTO) from a chemical enhancement perspective. The pyroelectric field generated by BaTiO3 drives the charge transfer between the SERS substrate and molecules, achieving a significant improvement in the SERS performance and self‐cleaning properties. The SERS signals of rhodamine 6G (R6G) molecules are further amplified up to 70‐fold. Thus, the detection limit is reduced by three orders of magnitude in this study, reaching 10−14 m after applying a pyroelectric field. In addition, the substrate exhibits a higher degradation efficiency than previous self‐cleaning SERS substrates because of the outstanding catalytic properties of BaTiO3. Target molecules are degraded effectively after several temperature cycles. A detailed mechanism analysis of pyroelectric SERS is conducted based on theoretical simulations and experimental results. This study is considered to deepen the understanding of SERS mechanisms and boost the application of SERS technology.

A Novel Fluorescent Chemosensor for Sensitive and Rapid Determination of Pb(II) Ions in Aqueous Environments.

This investigation devised an eco-friendly spectrofluorometric Pb(II) ion measurement approach. To determine Pb(II) ions, a polymeric fluorescence sensor membrane with excellent selectivity has been developed. It contains photoinitiators, functional monomers that can be cured under UV radiation, and cross-linking agents. After the characterization of the membrane, the most suitable parameters for the measurement of Pb(II) ions were determined. The measurement was performed in a short time-roughly 40s-at emission and excitation wavelengths of 488nm and 277nm, respectively, at pH 6.0. The developed method's detection limit was calculated as 1.42 × 10- 9 mol L- 1, and its calibration range was recorded as 4.83 × 10- 9 to 9.66 × 10- 8 mol L- 1. Recovery studies and the sensor's stability, lifetime, and repeatability have been evaluated. Real sample applications have been implemented using wastewater samples. The suggested approach may be used to quickly and accurately determine Pb(II) ions at low concentrations with good selectivity and sensitivity.

Detection of Ferric Ion by Fluorescent Carbon Nano Dots Synthesized from Forsythia Residue.

To fully utilize the wastes of the traditional Chinese herbs, a highly functionalized fluorescent carbon nano dots (CDs) based ferric ion sensor was prepared from forsythia residue via a one-step hydrothermal method. Under transmission electron microscope (TEM), the CDs were observed to be spherical with the diameter in the range of 5-20nm. Comprehensive analyses documented the CDs' favorable morphology, diverse functional groups, high water solubility, remarkable optical properties, and exceptional stability under various environmental conditions. Moreover, the CDs exhibited good optical properties with vivid green photoluminescence (PL) when they were exposed to ultraviolet (UV) light. Furthermore, the prepared CDs demonstrated selective fluorescence quenching behavior towards ferric ions with satisfactory sensitivity and a low limit of detection (LOD) of 4.3 µM. Additionally, the CDs displayed good selectivity towards Fe3+ and the least interference with several other metal ions. Consequently, this strategy could be effectively applied to real water samples, demonstrating its potential for broader applications.

A Multi-Channel Handheld Diagnostic Device Based on Green Biomimetic Material for Point-of-Care Testing of Vitamin C in Human Fluids.

In the era of "Great Health", maintaining an appropriate level of vitamin C is crucial for human health as it directly impacts the proper functioning of the immune system. In this study, a handheld nonenzymatic electrochemical sensor based on biomimetic material prussian blue functionalized polydopamine is proposed for point-of-care testing (POCT) of vitamin C in human fluids. Particularly, Prussian blue, known as a bio-antidote, and polydopamine, a main component of cuttlefish ink, ensure the safety and reliability of home testing. The experimental results quantitatively demonstrate that this electrochemical POCT sensor enables rapid and accurate determination of vitamin C with a wide linear detection range from 0.2 to 200µm, a low limit of detection at 0.03µm, and high sensitivity at 0.33µAµm-1cm-2. More importantly, the performance evaluation using human urine and saliva samples confirms satisfactory accuracy and recovery rates for vitamin C determination by this electrochemical POCT sensor. Thus, the proposed handheld electrochemical POCT sensor holds potential utility as an affordable tool for VC determination and home-based healthcare.

Easy-to-Engineer Flexible Nanoelectrode Sensor from an Inexpensive Overhead Projector Sheet for Sweat Neuropeptide-Y Detection.

In this paper, we report an inexpensive and easy-to-engineer flexible nanobiosensor electrode platform by exploring a nonconductive overhead projector (OHP) sheet for sweat Neuropeptide-Y (NPY) detection, a potential biomarker for stress, cardiovascular regulation, appetite, etc. We converted a nonconductive OHP sheet into a conductive nanobiosensor electrode platform with a hybrid polymerization method, which consists of interfacial polymerization of pyrrole and a template-free electropolymerization technique to decorate the electrode platform with poly(EDOT-COOH-co-EDOT-EG3) nanotubes. The selection of poly(EDOT-COOH) features an easy conjugation of NPY antibody (NPY-Ab) through EDC/Sulfo-NHS coupling chemistry, while poly(EDOT-EG3) is best known to reduce nonspecific binding of biomolecules. The antibody conjugation on the polymer surface was characterized by a quartz crystal microbalance, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and chronoamperometry techniques. The OHP nanosensor platform exhibited the successful detection of NPY analyte through a chronoamperometry method in phosphate-buffered saline with a wide range of concentrations from 1 pg/mL to 1 μg/mL with a limit of detection of 0.68 pg/mL having good linearity (R2 = 0.9841). The sensor platform exhibited excellent stability, reproducibility, repeatability, and a shelf-life of 13 days. Furthermore, the sensor showed superior selectivity to a 100 pg/mL NPY analyte among other interfering compounds such as tumor necrosis factor α, cortisol, and Interleukin-6. The clinical practicality of the sensor was confirmed through the detection of 100 pg/mL NPY spiked artificial perspiration, highlighting the possibility of integrating the sensor platform to wearable healthcare applications.

Carbon dot-graphene oxide-based luminescent nanosensor for creatinine detection in human urine.

Afluorescence (FL)-based nanosensor has been devised for creatinine (CR) detection in human urine specimens. The proposed nanosensor utilized a nanocomposite (NC) of carbon dots (CDs) and graphene oxide (GO). The formation of CDs/GO NC reduced the CD FL emission (λexcitation = 390nm, λemission = 461nm) by ~ 75%. Withthe introduction of CR to the NC, the CD emission intensity was reinstated by approximately 70%. The linear detection range for CR was 10-5 to 0.1mg dL-1 (R2 = 0.998), with a limit of detection of 4.3 × 10-2mg dL-1. Additionally, CDs/GO NC exhibited outstanding consistency and specificity in recognizing CR within urine specimens from both healthy individuals and patients suffering from chronic kidney disease(CKD). The Bland-Altman assessment (utilizing 25 human urine specimens) displayed remarkable consensus (R2 = 0.995) among theFL approach and the benchmark Jaffe technique. This observation indicates the hands-on usefulness of thenanosensor for identifying CR in biological specimens.

Dark Current Suppression in Two‐dimensional Histamine Lead Iodine Perovskite Single Crystal for X‐ray Detection and Imaging

AbstractLead halide perovskites have emerged as attractive X‐ray detector materials, owing to properties such as strong X‐ray stopping power, excellent carrier transport, and high sensitivity. Additionally, they can be easily prepared by using solution‐based synthesis approaches. However, traditional 3D (three‐dimensional) perovskites X‐ray detectors have shown limited application due to high dark currents generated under bias voltage as a result of strong ion migration. In this work, an X‐ray detector with a vertical structure device is demonstrated using 2D (two‐dimensional) histamine lead halide perovskite single crystal HAPbI4 (HPI, HA = histamine). Due to the dielectric screening effect of diamine and the vertical structure of the HPI device, the fabricated detector shows a sensitivity of 7737 µC Gyair−1 cm−2 under a bias voltage of 30 V. Furthermore, the detector shows a sensitivity of 293 µC Gyair−1 cm−2 and detection limit of 51.38 nGyair s−1 without bias voltage, wherein the dark current is almost completely suppressed. All of these properties indicate the X‐ray detection device is a promising candidate for next‐generation optoelectronic applications.

Development and Validation of a Stability Indicating Analytical Method for the Estimation of Gliclazide and Sitagliptin in Bulk Drugs and Tablet Dosage Forms by RP-UPLC: An Application of the Quality-by-Design Approach

Sitagliptin and gliclazide are the active ingredients in OpdualagTM. A highly selective DPP-4 inhibitor, sitagliptin is thought to work in type 2 diabetics by delaying the inactivation of incretin hormones, which increases their concentration and lengthens their duration of effect. Sulfonyl urea mostly enhances insulin secretion from pancreatic β-cells by inhibiting ATP-sensitive K+ channels, leading to depolarisation and Ca2+ influx. Reduced serum glucagon levels and enhanced insulin binding to receptors and target tissues are the mechanisms by which it exerts its effects. Objective: This study aims to create and evaluate an RP-UPLC technique for the detection of sitagliptin and gliclazide in pharmaceutical products by applying AQbD principles. Method: Findings By utilising the Design-expert® programme, we were able to execute a central composite design (CCD) consisting of three components arranged in five distinct layers, which greatly improved the chromatographic conditions. One method that oversees the whole analytical procedure life cycle—from method design and optimisation to validation and continual improvement—is Analytical Quality by Design (AQbD). Results: The retention times for sitagliptin were 1.269 minutes and for gliclazide they were 1.572 minutes. The limits of detection (LOD) and quantitation (LOQ) for Sitagliptin and Gliclazide were 0.19 µg/mL, 0.56 µg/mL, 0.28 µg/mL, and 0.86 µg/mL respectively. The recovery percentages for sitagliptin and gliclazide were found to be 99.15% to 100.54% and 99.93% to 100.58%, respectively. Both drugs were found to be susceptible to oxidative and photolytic breakdown in the forced degradation studies. Conclusions A novel RP-UPLC method has been developed for the quantitative detection of sitagliptin and gliclazide in pharmaceutical formulations, utilising the AQbD-based methodology. This approach is straightforward, rapid, precise, particular, and stable-indicating.

Detection of "smoke powder" etomidate and its metabolite etomidate acid in blood and urine by UHPLC-MS-MS: application in authentic cases.

Recently, etomidate has been widely used as an alternative in illicit drug market. It is usually added to regular cigarette tobacco (commonly known as "cigarette powder") or mixed in e-cigarette oil sold through the Internet, retail stores, or entertainment outlets and other channels. An ultra-high performance liquid chromatography tandem mass spectrometry method was developed to quantify etomidate and etomidate acid in human blood and urine. The limit of detection (LOD) of etomidate and etomidate acid in blood is 0.5 and 2 ng/mL, respectively, and the lower limit of quantification (LLOQ) is 1 and 5 ng/mL, respectively. The LOD of etomidate and etomidate acid in urine is 1 and 2 ng/mL, respectively, and the LLOQ is 2 and 5 ng/mL, respectively. The precision, accuracy, recoveries, and matrix effects of etomidate and etomidate acid determinations in blood and urine met the requirements for methodological validation. The method was successfully applied to the identification and quantification of etomidate and etomidate acid in blood and urine of 62 forensic cases. The concentration of etomidate ranged from 1.52 to 8.41 ng/mL (positive cases, n = 5) and the concentration of etomidate acid ranged from 2.76 to 112 ng/mL (positive cases, n = 5) in blood. The concentrations of etomidate and etomidate acid in urine samples were 2.64-79,300 ng/mL (positive cases, n = 59) and 6.11-518,000 ng/mL (positive cases, n = 60), respectively. Therefore, the concentration of etomidate in blood and urine is mostly higher than that of etomidate.

Compact highly sensitive photothermal RT-LAMP chip for simultaneous multidisease detection.

Developing instant detection systems with disease diagnostic capabilities holds immense importance for remote or resource-limited areas. However, the task of creating these systems-which are simultaneously easy to operate, rapid in detection, and cost-effective-remains a challenge. In this study, we present a compact highly sensitive photothermal reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) chip (SPRC) designed for the detection of multiple diseases. The nucleic acid (NA) amplification on the chip is achieved through LAMP driven by either LED illumination or simple sunlight focusing. SPRC performs sample addition and amplification within a limited volume and autonomous enrichment of NA during the sample addition process, achieving a limit of detection (LOD) as low as 0.2 copies per microliter. Through 120 clinical samples, we achieved an accuracy of 95%, with a specificity exceeding 97.5%. Overall, SPRC has achieved promising progress in the application of point-of-care testing (POCT) by using light energy to simultaneously detect multiple diseases.

Precise Genotyping Via Surface‐Enhanced Raman Spectroscopy‐Based Optical Sensing Chip for Guiding Targeted Therapy in Lung Cancer

AbstractThe emergence of “precision medicine” marks a notable shift in cancer treatment, moving from a tumor type–oriented approach to a more targeted, gene‐oriented approach. Detecting low‐abundance mutant genes in blood is challenging but crucial for personalized treatment plans. Herein, a novel platform combining catalytic hairpin self‐assembly (CHA)‐mediated self‐calibrating surface‐enhanced Raman spectroscopy (SERS) with a high‐throughput Raman system (CCSPS) was designed. This platform enables ultrasensitive and rapid genotype analysis of gene mutations. The development of CCSPS specifically targets EGFR mutations, which serve as crucial therapeutic targets for precision therapy in lung cancer. This system shows excellent sensitivity and selectivity, capable of detecting multiple EGFR mutations (Del‐19, L858R, and T790M) with a detection limit as low as attomolar levels. Additionally, precise genotyping analysis was successfully conducted on 42 clinical samples using the CCSPS, yielding results consistent with those obtained through next‐generation sequencing. These results underscore the efficacy of the CCSPS in noninvasively identifying circulating tumor DNA (ctDNA) mutations, facilitating immediate therapeutic decision making at the bedside. In summary, the CCSPS is a fast, accurate, versatile, and compact testing system capable of precisely screening individuals who stand to benefit from targeted therapy, thus promoting personalized and precise healthcare.

Quantum dot-based biomimetic fluorescence immunoassays for enrofloxacin detection in animal-derived foods.

Long-term consumption of foods with excessive enrofloxacin (ENRO) residues may cause the accumulation of ENRO in the human body, thus damaging human health. In this study, quantum dot-based biomimetic fluorescence immunoassays were used for enrofloxacin detection in food of animal origin. Under the most suitable conditions, the detection limit (IC15) of the method in standard solution was 0.13 ± 0.02 ng L-1 and the sensitivity (IC50) was 0.13 ± 0.18 μg L-1. The recoveries for ENRO from four fortified food samples, including chicken, eggs, shrimp, and milk, ranged from 85.74% to 114.19% and the coefficients of variation were 0.01-18.09%. The established method shows good agreement with the results obtained using commercial ELISA kits, with a correlation coefficient of 0.997. The proposed method shows the advantages of high sensitivity, specificity and wide detection range. It can be used as an alternative method for the rapid and sensitive detection of ENRO in food of animal origin.

Eco-Friendly Synchronous Spectrofluorimetric Determination of Imipenem, Cilastatin, and Relebactam; Application to Market Formulations and Biological Fluids; Greenness Assessment.

The proposed study introduces a rapid, sensitive, and simple synchronous spectrofluorimetric technique for simultaneous quantification of relebactam, cilastatin, and imipenem in marketed pharmaceutical forms and biological fluids. Using synchronous fluorescence spectroscopy at Δ λ = 110nm, cilastatin was detected at 360nm. Fourier Self-Deconvolution was subsequently applied to the spectrum to estimate relebactam and imipenem at 430nm and 470nm, respectively after detection of cilastatin at 360nm ensuring no cross-interference. The pH was adjusted to 8.0 using 2.0 mL of alkaline borate buffer. This approach allowed for the precise quantification of relebactam, cilastatin, and imipenem through ranges of 50-400 ng mL- 1, 20-500 ng mL- 1, and 50-500 ng mL- 1 respectively. The lower detection and quantitation limits were 9.9 and 29.7 ng mL- 1 for REL, 4.5 and 13.6 ng mL- 1 for CIL and 5.5 and 16.5 ng mL- 1 for IMP. The proposed method was successfully applied for the determination of studied drugs in their pharmaceutical formulations with a high degree of accuracy and without interference from common excipients. This approach allowed for the precise quantification of relebactam, cilastatin, and imipenem through ranges of 50-400 ng mL- 1, 20-500 ng mL- 1, and 50-500 ng mL- 1, respectively. The proposed method was rigorously validated according to ICH guidelines. Furthermore, the method's environmental impact was assessed using Eco-scale and Green Analytical Procedure Index (GAPI) techniques.

The AstroSat UV Deep Field South. I. Far- and Near-ultraviolet Source Catalog of the GOODS South Region

We present the AstroSat UV Deep Field south (AUDFs), an imaging survey using the wide-field Ultraviolet Imaging Telescope on board AstroSat. AUDFs cover ∼236 arcmin2 of the sky area, including the Great Observatories Origins Deep Survey (GOODS) South field in F154W and N242W filters. The deep and shallow parts of AUDFs have an exposure time ∼62,000 and ∼31,000 s, respectively, in the F154W filter, while in the N242W filter, they are ∼64,000 and ∼34,000 s. These observations reached a 3σ depth of 27.2 and 27.7 AB mag with a 50% completeness limit of 27 and 27.6 AB mag in the F154W and N242W filters, respectively. With the acquired depth, AUDFs is the deepest far- and near-UV imaging data covering the largest area known to date at 1.″2–1.″6 spatial resolution. Two primary catalogs were constructed for the F154W and N242W filters, each containing 13,495 and 19,374 sources brighter than the 3σ detection limit, respectively. Our galaxy counts of a power-law slope of ∼0.43 dex mag−1 in the N242W filter match well with Hubble Space Telescope/Wide-Field Camera 3/UVIS observations. A wide range of extragalactic science can be achieved with this unique data, such as providing a sample of galaxies emitting ionizing photons in the redshift range of z ∼1–3 and beyond, constraining the UV luminosity function, investigating the extended UV emission around star-forming galaxies and UV morphologies for z < 1. The UV catalog will enhance the legacy value of the existing optical/IR imaging and spectroscopic observations from ground- and space-based telescopes on the GOODS South field.

Ion‐Selective Mobility Differential Amplifier: Enhancing Pressure‐Induced Voltage Response in Hydrogels

Piezoionics is an emerging mechanical‐electrical energy conversion paradigm that enables self‐powered sensing systems for next‐generation intelligent wearable electronics. However, there are currently no rational design approaches to enhance the stimulus response of piezoionic devices. Here, we present a strategy using crown ether as ion‐selective mobility differential amplifiers for enhancing the pressured‐induced voltage response in ionic polyvinyl alcohol (PVA) hydrogels. The crown ether grafted PVA (PVA‐CE) hydrogel prototype achieves a 30‐fold amplified piezoionic coefficient of 1490 nV Pa−1 within 0 – 1 kPa, compared to 49 nV Pa−1 of the unmodified PVA. The PVA‐CE exhibits an ultra‐low pressure detection limit of 0.2 Pa with a fast response time of 18.1 ms. Leveraging these properties, we further demonstrate a human somatosensory network analogous PVA‐CE piezoionic skin using arrayed pressure sensing. These capabilities hold great promise for emerging healthcare applications such as synthetic biology, soft robotics, and beyond.

Rapid on-site detection of viable Vibrio parahaemolyticus in seafood using cis-diamminedichloroplatinum and colorimetric loop-mediated isothermal amplification (CDDP-LAMP).

Vibrio parahaemolyticus in seafood and marine environments poses significant health risks, causing gastroenteritis worldwide. Current detection methods fail to differentiate live from dead cells, leading to inaccuracies in food safety assessments. This study introduces a novel method combining cis-diamminedichloroplatinum (CDDP) with direct colorimetric loop-mediated isothermal amplification (LAMP) for rapid and accurate detection of viable V. parahaemolyticus cells in seafood samples. CDDP treatment at 37°C for 30min selectively inhibits DNA from dead cells, enhancing the specificity of the assay by ensuring only live cell DNA is amplified. The optimized CDDP-LAMP procedure detects alive V. parahaemolyticus within 1h, with results observable through a color change. The CDDP-LAMP assay demonstrates excellent specificity, identifying live V. parahaemolyticus cells while excluding dead cells and other bacteria. It shows a detection limit of 2.348CFU per reaction and successfully detects V. parahaemolyticus in seafood samples across different food matrices. This study is the first to combine CDDP with colorimetric LAMP for direct detecting viable bacteria in food, enhancing specificity by eliminating signals from dead cells. The CDDP-LAMP assay provides a rapid, accurate process for detecting viable V. parahaemolyticus cells, especially in resource-limited settings. It also gives a model for screening different bacterial pathogens, speeding up and improving foodborne illness risk assessments.

Optimization of the Combined Use of Z-Sep Plus and EMR-Lipid in QuEChERS Procedure for the Analysis of Eight Pesticides in Real Milk Samples

AbstractOne of the most applied procedures for the determination of trace analytes in complex matrices is QuEChERS (an acronym for Quick, Easy, Cheap, Effective, Rugged, and Safe). QuEChERS procedures include an extraction step followed by a dispersive solid-phase extraction (dSPE) for analytes cleaning-up from the matrix components. A challenging task in QuEChERS procedures is extracting and determining pesticides from samples of high fat such as milk samples. This challenge induced the innovation of new adsorbents for the clean-up step such as Z-Sep Plus® and EMR-Lipid® to enable removal of fatty matrix components without affecting the recovery of hydrophobic analytes. This work aims to apply experimental design to optimize the combined application of both QuEChERS clean-up adsorbents; Z-Sep Plus® and EMR-Lipid® in addition to other QuEChERS parameters in the determination of eight pesticides: hexachlorocyclohexane, dichlorodiphenyldichloroethane, dichlorodiphenyltrichloroethane, primiphos ethyl, diazinon, malathion, endrin, and dimethoate in milk matrix. This was augmented by optimization of GC–MS/MS and UPLC-MS/MS to detect and determine analytes in extracts. The experimental design of QuEChERS procedure enabled the optimization of Z-Sep Plus®- and EMR-Lipid®-added adsorbent amounts with other method parameters to enable the maximum recovery of analytes. Furthermore, the optimized methods enabled low detection limits of the studied pesticides within a short analysis time (28 min for GC and 12 min for LC methods, respectively). The procedure was validated according to European SANTE/11312/2021 Guideline. Quantitation limit ranged from 1.7 to 3.2 ng/mL for GC–MS/MS method and from 1.7 to 3 ng/mL for UPLC-MS/MS method. Greenness assessment of the methods followed four approaches indicating an excellent value of greenness for the proposed methods. Furthermore, 45 real milk samples collected from the Egyptian market were tested with the developed procedure for the presence of pesticides.