Sensitive Method Research Articles

Pharmacokinetic study of iptacopan and its two acyl glucuronide metabolites in monkey plasma by liquid chromatography combined with electrospray ionization tandem mass spectrometry.

In this study, a simple and sensitive liquid chromatography tandem mass spectrometric method was developed and validated for the determination of iptacopan and two acyl glucuronidation metabolites in monkey plasma. The plasma sample was precipitated with acetonitrile and then separated on an Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm) using 0.1% formic acid and 5 mM ammonium acetate in water and acetonitrile as the mobile phase. The mass spectrometry (MS) detection was performed in positive multiple reactions monitoring (MRM) mode with precursor-to-production transitions. The developed assay was validated over the range of 1-2000 ng/mL for three analytes with correlation coefficient (r) more than 0.99. The validation parameters including accuracy, precision, carryover effect, matrix effect, recovery, and stability were all within the acceptable limits. The validated method has been applied to investigate the pharmacokinetics of iptacopan and its two acyl glucuronidation metabolites in monkey plasma. After intravenous administration, iptacopan showed low clearance (2.75 mL/min/kg) in monkey plasma. After oral administration, the bioavailability was 55.43%. The exposure (AUC0-t) of direct acyl glucuronide (AG) of iptacopan accounts for 9.73% of the iptacopan plasma exposure. The AUC0-t of AG of dealkylated metabolite of iptacopan was present at a lower level, accounting for 0.5% of the iptacopan plasma exposure.

A Quantitative Real-Time PCR Method to Detect the Quinoa Downy Mildew Pathogen, Peronospora variabilis.

Quinoa downy mildew, caused by Peronospora variabilis, is the most devastating disease of quinoa globally. Rapid, sensitive diagnostic methods are needed to detect and quantify this pathogen in seeds and plant tissue. A hydrolysis probe-based quantitative real-time PCR (qPCR) assay including a competitive internal control was developed for P. variabilis detection. This assay could detect as low as 20 ag of DNA or approximately 25 internal transcribed spacer (ITS) copies per reaction with efficiencies ranging from 93.9 to 98.2%. No nontarget amplification was observed when tested against DNA from other downy mildew pathogens and related oomycetes. P. variabilis strains from multiple countries were detected using this assay. The assay was successfully applied to quantify the pathogen in quinoa seeds from a field trial conducted in the state of Washington. Downy mildew disease was recorded on all 14 genotypes, with the genotypes 104.88 and 106.49 recording the highest area under the disease progress curve values (mean ± SE; 3,236 ± 303 and 2,851 ± 198, respectively) and J6 and Dutchess recording the lowest (441 ± 107 and 409 ± 129, respectively). Seed washes obtained from field samples were subjected to the qPCR assay, and the pathogen was detected in all samples. The highest pathogen ITS copy number was recorded with 106.49 (194,934 ± 38,171), and the lowest was observed in Pasto (5,971 ± 1,435) and Riobamba (9,954 ± 4,243). This qPCR assay could lead to improved detection and quantification of P. variabilis as well as increased understanding of quinoa-P. variabilis interactions and epidemiology.

A sensitive and robust analytical method for the determination of enramycin residues in swine tissues using UHPLC-MS/MS.

Enramycin, a common growth promoter utilized in chickens and pigs, is sensitive against Gram-positive bacteria, and the maximum residue limit (MRL) of enramycin set up by is 30 μg/kg. However, the methods have been reported for detecting enramycin have failed to meet the accuracy requirements, with the required limit of quantification being higher than the MRL. To address this issue, we developed a high-sensitive and robust analytical method based on ultrahigh-performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS), to determine enramycin residues in swine tissues, including liver, kidney, pork, and fat. The ENV cartridge was selected to cleanup and enrich analytes after being extracted using a mixture of 55% methanol containing 0.2 M hydrochloric acid. With comprehensively validation, this established method was found great linearity of enramycin in each tissue, with a coefficient of variation above 0.99. Satisfactory recoveries from four different spiking levels were acquired (70.99-101.40%) while the relative standard deviations were all below 9%. The limit of quantification of enramycin in the present study is 5 μg/kg in fat and 10 μg/kg in other tissues, meeting the requirements for conducting the corresponding safety evaluation study. This method was demonstrated with excellent specificity, stability, and high sensitivity. To conclude, this novel approach is sufficiently sensitive and robust for the safety evaluation of enramycin in food products.

A novel system with robust compatibility and stability for detecting Sugarcane yellow leaf virus based on CRISPR-Cas12a.

Sugarcane yellow leaf virus (SCYLV) can reduce sugarcane productivity. A novel detection system based on reverse transcription-multienzyme isothermal rapid amplification (RT-MIRA) combined with CRISPR-Cas12a, named RT-MIRA-CRISPR-Cas12a, was developed. This innovative approach employs crude leaf extract directly as the reaction template, streamlining the extraction process for simplicity and speed. Combining RT-MIRA and CRISPR-Cas12a in one reaction tube increases the ease of operation while reducing the risk of aerosol contamination. In addition, it exhibits sensitivity equivalent to qPCR, boasting a lower detection limit of 25 copies. Remarkably, the entire process, from sample extraction to reaction completion, requires only 52-57 minutes, just a thermostat water bath. The result can be observed and judged by the naked eye.IMPORTANCESugarcane yellow leaf disease (SCYLD) is an important viral disease that affects sugarcane yield. There is an urgent need for rapid, sensitive, and stable detection methods. The reverse transcription-multienzyme isothermal rapid amplification combined with CRISPR-Cas12a (RT-MIRA-CRISPR-Cas12a) method established in this study has good specificity and high sensitivity. In addition, the system showed good compatibility and stability with the crude leaf extract, as shown by the fact that the crude extract of the positive sample could still be stably detected after 1 week when placed at 4°C. RT-MIRA-CRISPR-Cas12a, reverse transcription polymerase chain reaction (RT-PCR), and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to detect SCYLV on 33 sugarcane leaf samples collected from the field, and it was found that the three methods reached consistent conclusions. This Cas12a-based detection method proves highly suitable for the rapid on-site detection of the SCYLV.

2D/0D Heterojunction Fluorescent Probe with Schottky Barrier Based on Ti3C2TX MXene Loaded Graphene Quantum Dots for Detection of H2S During Food Spoilage

AbstractHydrogen sulfide (H2S) contamination of food has raised widespread public health concerns, leading to substantial medical and economic burdens. Herein, a 2D/0D heterojunction fluorescent probe (TCTG) with Schottky barriers (SB) is designed and synthesized, utilizing Ti3C2Tx MXene‐loaded graphene quantum dots (GQDs), for the detection of H2S during food spoilage. The microstructures observed through SEM and TEM reveal that uniformly sized GQDs are evenly attached to the surface of a monolayer Ti3C2Tx. The chemisorption between GQDs and Ti3C2Tx facilitates charge transfer and the formation of SB, resulting in intramolecular charge transfer (ICT) effects. With the introduction of H2S, TCTG(50%) exhibits the highest sensitivity, selectivity, and anti‐interference properties, with ultra‐fast fluorescence transient reaction (3s) and remarkably low detection limit of 41.82 ppb as well as noticeable color change. When TCTG(50%) reacted with H2S, the ICT effects are inhibited, leading to the recovery of photoinduced electron transfer (PET) and fluorescence quenching. Notably, probe TCTG is effectively utilized to detect changes in H2S levels in raw foods to assess their quality. Overall, the significance of this study is its potential to revolutionize food spoilage detection, offering a fast, reliable, and sensitive method to ensure food safety and reduce associated health and economic burdens.

The use of an RP-HPLC-UV method for the analysis of oxcarbazepine in the presence of its preservatives; stability studies and application to human plasma samples

AbstractA simple, sensitive, selective, accurate and precise method was developed and fully validated for determination of oxcarbazepine (OXC) in presence of their preservatives and determination of oxcarbazepine (OXC) in human plasma. A reversed phase liquid chromatography (RP-HPLC) with UV detection techniques were applied for separation and quantification of studied drug OXC. Successful separation of the drug from methyl paraben (M.P.), propyl paraben (P.P.) and potassium sorbate (P.ST.) was achieved on a Kromasil C18 column (5 μm particle size, pore size 300 Å, l × I.D. 250 × 4.6 mm). The mobile phase that contain aqueous 0.05M potassium dihydrogen phosphate buffer (pH 7): acetonitrile, (50: 50, %v/v). The method was linear over concentration ranges 5.0–50 μg mL−1 for OXC. Bioanalytical validation of the developed method was carried out according to US-FDA guidelines and revealed a good linear relations over a range of (5.0–50), (0.5–10), (0.05–0.15), and (1.0–10) μg mL−1 for OXC, M.P, P.P, and P.ST, respectively, with a correlation coefficient (R2) of more than 0.999. Limit of detection (LOD) were 1.15, 0.03, 0.01 and 0.04 μg mL−1 for OXC, M.P, P.P, and P.ST, respectively, Intra and inter-day precisions, calculated as percentage relative standard deviation (% RSD), were lower than 2.0%. The developed method can be applied for routine drug analysis, therapeutic drug monitoring and bioequivalence studies through the analysis of plasma samples taken from blood bank.

Development and validation of an LC-MS/MS method for the quantification of free and total doxorubicin in human plasma and its clinical application for a novel doxorubicin hydrochloride liposome injection in Chinese patients with breast cancer

AbstractA novel doxorubicin hydrochloride liposome injection was prepared to reduce toxicity and side effects, as well as extend plasma half-life in the treatment of breast cancer. In this study, a rapid and sensitive bioanalytical method was developed and validated to characterize the pharmacokinetic profile of total and free doxorubicin in plasma of 3 Chinese patients after intravenous infusion of this injection. Plasma samples were prepared by protein precipitation for the determination of total doxorubicin, while solid phase extraction was used to determine free doxorubicin. After plasma sample pre-treatment, total and free concentrations were quantified individually using a validated LC-MS/MS method. The calibration curves were found to be linear in the range of 0.20–500.0 ng mL−1 for total doxorubicin and in the range of 1.00–1,000 ng mL−1 for free doxorubicin. The free concentrations in plasma were only one sixth to one quarter of the total levels. Liposomal doxorubicin had a longer apparent half-life (>50 h) than the non-targeted drug (<10 h) reported in the reference. and a lower volume of distribution. This novel injectable formulation steadily released free doxorubicin from liposomes over a long period of time to reduce cardiac toxicity and side effects, while ensuring a clinical curative effect.

Combining Fourier-transform infrared spectroscopy and multivariate analysis for chemotyping of cell wall composition in Mungbean (Vigna radiata (L.) Wizcek)

BackgroundDissection of complex plant cell wall structures demands a sensitive and quantitative method. FTIR is used regularly as a screening method to identify specific linkages in cell walls. However, quantification and assigning spectral bands to particular cell wall components is still a major challenge, specifically in crop species. In this study, we addressed these challenges using ATR-FTIR spectroscopy as it is a high throughput, cost-effective and non-destructive approach to understand the plant cell wall composition. This method was validated by analysing different varieties of mungbean which is one of the most important legume crops grown widely in Asia.ResultsUsing standards and extraction of a specific component of cell wall components, we assigned 1050–1060 cm−1 and 1390–1420 cm−1 wavenumbers that can be widely used to quantify cellulose and lignin, respectively, in Arabidopsis, Populus, rice and mungbean. Also, using KBr as a diluent, we established a method that can relatively quantify the cellulose and lignin composition among different tissue types of the above species. We further used this method to quantify cellulose and lignin in field-grown mungbean genotypes. The ATR-FTIR-based study revealed the cellulose content variation ranges from 27.9% to 52.3%, and the lignin content variation ranges from 13.7% to 31.6% in mungbean genotypes.ConclusionMultivariate analysis of FT-IR data revealed differences in total cell wall (600–2000 cm−1), cellulose (1000–1100 cm−1) and lignin (1390–1420 cm−1) among leaf and stem of four plant species. Overall, our data suggested that ATR-FTIR can be used for the relative quantification of lignin and cellulose in different plant species. This method was successfully applied for rapid screening of cell wall composition in mungbean stem, and similarly, it can be used for screening other crops or tree species.

Study of the Synchronization and Transmission of Intracellular Signaling Oscillations in Cells Using Bispectral Analysis

The oscillation synchronization analysis in biological systems will expand our knowledge about the response of living systems to changes in environmental conditions. This knowledge can be used in medicine (diagnosis, therapy, monitoring) and agriculture (increasing productivity, resistance to adverse effects). Currently, the search is underway for an informative, accurate and sensitive method for analyzing the synchronization of oscillatory processes in cell biology. It is especially pronounced in analyzing the concentration oscillations of intracellular signaling molecules in electrically nonexcitable cells. The bispectral analysis method could be applied to assess the characteristics of synchronized oscillations of intracellular mediators. We chose endothelial cells from mouse microvessels as model cells. Concentrations of well-studied calcium and nitric oxide (NO) were selected for study in control conditions and well-described stress: heating to 40 °C and hyperglycemia. The bispectral analysis allows us to accurately evaluate the proportion of synchronized cells, their synchronization degree, and the amplitude and frequency of synchronized calcium and NO oscillations. Heating to 40 °C increased cell synchronization for calcium but decreased for NO oscillations. Hyperglycemia abolished this effect. Heating to 40 °C changed the frequencies and increased the amplitudes of synchronized oscillations of calcium concentration and the NO synthesis rate. The first part of this paper describes the principles of the bispectral analysis method and equations and modifications of the method we propose. In the second part of this paper, specific examples of the application of bispectral analysis to assess the synchronization of living cells in vitro are presented. The discussion compares the capabilities of bispectral analysis with other analytical methods in this field.

Genetic evolutionary analysis of a strain of Senecavirus A in Anhui and the establishment of its detection method

BackgroundSenecavirus A (SVA) is the only member of the genus Senecavirus in the family Picornaviridae, and is one of the pathogens of porcine blistering disease. SVA has been reported in the United States, Canada, China, Thailand, and Colombia. MethodsIn this study, positive SVA infection was detected by RT-PCR in sick materials collected from pig farms of different sizes in Anhui Province. ResultsIn this study, a virulent strain of SVA was successfully obtained by viral isolation on BHK21 cells and named SVA-CH-AHAU-1. Meanwhile, a simple, rapid and accurate nano-PCR method for the detection of SVA infection was established in this study, using the recombinant plasmid pClone-SVA-3D as a template. ConclusionsThe complete genome of SVA-CH-AHAU-1 is 7286 bp, including a 5′ non-coding region (UTR), an open reading frame (ORF) of 6546 nucleotides, encoding 2182 amino acids (aa), and a 3’ UTR with Poly(A) features, and phylogenetic analysis showed that this isolate had the highest nucleotide homology (97.9 %) with the US isolate US-15-41901SD. In this study, the virulent strain SVA-CH-AHAU-1 was found to recombine in the ORF region with isolates SVA-CH-SDGT-2017 and SVA/Canada/ON/FMA-2015-0024 T2/2015. The complete genome has been submitted to GeneBank with the accession number OM654411. In addition, our results suggest that the established nano-PCR assay can be used as an economical, reliable and sensitive method for the field diagnosis of SVA method, especially in resource-limited areas.

Simultaneous Determination of Tobacco Smoke Exposure and Stress Biomarkers in Saliva Using In-Tube SPME and LC-MS/MS for the Analysis of the Association between Passive Smoking and Stress.

Passive smoking from environmental tobacco smoke not only increases the risk of lung cancer and cardiovascular disease but may also be a stressor triggering neuropsychiatric and other disorders. To prevent these diseases, understanding the relationship between passive smoking and stress is vital. In this study, we developed a simple and sensitive method to simultaneously measure nicotine (Nic) and cotinine (Cot) as tobacco smoke exposure biomarkers, and cortisol (CRT), serotonin (5-HT), melatonin (MEL), dopamine (DA), and oxytocin (OXT) as stress-related biomarkers. These were extracted and concentrated from saliva by in-tube solid-phase microextraction (IT-SPME) using a Supel-Q PLOT capillary as the extraction device, then separated and detected within 6 min by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using a Kinetex Biphenyl column (Phenomenex Inc., Torrance, CA, USA). Limits of detection (S/N = 3) for Nic, Cot, CRT, 5-HT, MEL, DA, and OXT were 0.22, 0.12, 0.78, 0.39, 0.45, 1.4, and 3.7 pg mL-1, respectively, with linearity of calibration curves in the range of 0.01-25 ng mL-1 using stable isotope-labeled internal standards. Intra- and inter-day reproducibilities were under 7.9% and 14.6% (n = 5) relative standard deviations, and compound recoveries in spiked saliva samples ranged from 82.1 to 106.6%. In thirty nonsmokers, Nic contents positively correlated with CRT contents (R2 = 0.5264, n = 30), while no significant correlation was found with other biomarkers. The standard deviation of intervals between normal beats as the standard measure of heart rate variability analysis negatively correlated with CRT contents (R2 = 0.5041, n = 30). After passive smoke exposure, Nic levels transiently increased, Cot and CRT levels rose over time, and 5-HT, DA, and OXT levels decreased. These results indicate tobacco smoke exposure acts as a stressor in nonsmokers.

Understanding Latent Variables in EFL Contexts

Understanding latent variables is essential in EFL research. This article examines key latent variables, such as linguistic competence, cognitive ability and socio-cultural factors. These variables play a crucial role in shaping EFL learning experiences and outcomes. Researchers can use methods such as exploratory factor analysis (EFA), confirmatory factor analysis (CFA) and structural equation modeling (SEM) to uncover these variables. By combining theoretical findings with advanced statistical methods, such research provides an understanding of the complex processes involved in language learning. The article points to the need for longitudinal studies and mixed methods to capture the dynamic nature of EFL learning. This approach could contribute to the development of more effective educational strategies that improve teaching practices and learner outcomes. The article also emphasizes the importance of considering the sociocultural context and developing culturally sensitive teaching methods to create an inclusive learning environment. Moreover, this article serves as a guide for future research and practice in language learning.

One-pot derivatization spectrofluorimetric method for detection of aspartame in sweetener commercial tablets and soft drinks: Greenness assessments of the methodology.

Aspartame is an artificial sweetener used in drinks and many foods. International Agency for Research on Cancer classified aspartame as possibly carcinogenic to humans (IARC Group 2B). In this study, a sensitive and selective spectrofluorimetric method was developed to detect aspartame. The method is based on switching on the fluorescence activity of aspartame upon its condensation with O-phthalaldehyde (Roth's reaction) in the presence of 2-mercaptoethanol. The reaction product was detected fluorometrically at λem of 438 nm after λex of 340 nm. All reaction conditions required to yield the optimal fluorescence intensity were observed and investigated. Furthermore, the approach was validated according to ICH guidelines. Upon plotting the concentrations of aspartame against their associated fluorescence intensity values, the relationship between the two variables was linear within the range of 0.5-3.0μg/mL. Furthermore, the method was employed to analyze the quantity of aspartame in commercial packages and soft drinks with an acceptable level of recovery. In addition, the Green Solvents Selecting Tool, Complementary Green Analytical Procedure Index, and the Analytical Greenness Metric tool were used to evaluate the sustainability and the greenness of the developed methodology.

Blueness, Whiteness, and greenness Appraisal of sensitive chromatographic approach for determination of semaglutide concurrently with Pioglitazone and metformin using green mobile Phase; alongside with pharmacokinetic study

The replacement of hazardous solvents with greener alternatives was under consideration to achieve a sustainable and green approach. The assessment of reliable white analytical method for the determination of semaglutide as a novel member in glucagon-like peptide agonists class is still of a great importance. That is because pharmacological, clinical, and therapeutic drug monitoring purposes. This research proposes and fully validates a simple and sensitive RP-HPLC method for the determination of semaglutide simultaneously with pioglitazone and metformin. The suggested procedures utilizing a green mobile phase of 8 % glycerol in 0.01 M phosphate buffer (pH 6). Separation was achieved using a cyano (5.0 μm, 150 × 4.6 mm) column maintained at a temperature of 40 °C. The mixture was isocratically eluted under a flow rate of 1 mL/min with a total run time of less than 8 min. The UV-detector was adjusted at 215 nm. The method provides low detection (ranging from 0.003 to 0.63 µg/mL) and quantification (ranging from 0.008 to 1.9 µg/mL) limits, with good precision (%RSD ranging from 0.66 % to 0.89 %). The suggested technique has been effectively used to determine each drug in its pharmaceutical preparation and spiked human plasma. Beside the remarkable sensitivity and sustainability of the evolved approach, the relevance of therapeutic drug monitoring motivated us to perform a pharmacokinetics study for both semaglutide and metformin in real human plasma samples. Greenness, whiteness and blueness assessment of the developed method have been carried out using different tools (Green Analytical Procedure Index (GAPI), the analytical GREEnness metric approach (AGREE), RGB-12 model and BAGI). The proven green and white profile of the evolved method could open future prospective for the clinical research and regular quality control examination of the investigated mixture with the least negative environmental impacts.

Cobalt oxyhydroxide nanoflakes enable ratiometric fluorescent assay of gallic acid

Gallic acid (GA) is widely used in beverages, food, and other fields as antioxidant. However, GA is slightly toxic and the accumulation of GA is harmful to human body. Therefore, it's vital to develop simple and sensitive detection methods for GA. In this work, a novel ratiometric fluorescent nanoprobe (named CoOOH/OPD/SiNPs) for the GA detection in different foods was designed and prepared. The fluorescence of silicon nanoparticles (SiNPs) at 443 nm would be quenched by cobalt oxyhydroxide (CoOOH) nanoflakes. o-phenylenediamine (OPD) would be oxidized to 2,3-diaminophenazine (DAP) by CoOOH nanoflakes that have peroxidase-like activity, which produces a new fluorescent peak at 556 nm. Meanwhile, SiNPs' fluorescence would be quenched through DAP due to inner filter effect (IFE). With the addition of GA, the reductive decomposition of CoOOH decreased DAP level, causing IFE being restrained. The concentration of GA indicates an excellent linear relationship with fluorescence ratio (F443/F556) in range of 0.4–12 μM (R2 = 0.9937) with 0.16 μM detection limit. This nanoprobe is applied to GA detection in water, tea leaves, fruits and nut fruits, which would be expected to act as a portable device for complex substances analysis.

Zirconium-based metal-organic framework composites for solid phase extraction of brevetoxin-A from seawater

Red tides are a type of natural marine disaster caused by harmful algae characterized by a high toxicity, wide distribution, and long duration. Since the concentration of algal toxins in seawater increases with the occurrence of red tides, algal toxins detected in seawater could be used to predict the occurrence and evolution of red tides. Brevetoxin-A (BTX-A) is a secondary metabolite produced by the harmful algae Karenia brevis, whose detection in seawater could form the basis of an accurate warning system for incoming red tides. However, due to the inherent complexity of the seawater matrix and the extremely low levels of BTX-A in seawater, the use of instruments for its direct detection is difficult. Therefore, there is an urgent need to develop a sample pretreatment method for the efficient enrichment of BTX-A in seawater. In this study, a metal-organic backbone material (UiO-66) and its composite with silica microspheres (SiO2@UiO-66) were successfully synthesized using the solvothermal method. The prepared SiO2@UiO-66 exhibited good hydrophilicity, water stability, and large specific surface area. Furthermore, it also exhibited hydrogen bonding and electrostatic interactions with BTX-A, had a strong affinity for BTX-A, and was able to efficiently adsorb BTX-A in complex matrices. Therefore, SiO2@UiO-66 showed potential as a novel packing material for the extraction of BTX-A from solid phase extraction columns. Combined with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), a highly sensitive detection method for the determination of BTX-A in marine water was established. The established analytical method had a low detection limit (3.0 pg/mL), a wide linear range (10.0 -200.0 pg/mL), and a good linear relationship (R=0.9992). Combined with the Fujian Province Red Tide Monitoring and Early Warning Information 2021 issued by the Fujian Provincial Oceanic and Fisheries Bureau, the analytical method established herein was successfully applied to analyze and monitor the content of BTX-A in actual seawater samples. This highlights the proposed system's potential for use as an early warning factor in the monitoring of red tides, representing a simple and fast pretreatment methodology for the detection of BTX-A in seawater.

A Machine Learning-Driven Surface-Enhanced Raman Scattering Analysis Platform for the Label-Free Detection and Identification of Gastric Lesions.

Gastric lesions pose significant clinical challenges due to their varying degrees of malignancy and difficulty in early diagnosis. Early and accurate detection of these lesions is crucial for effective treatment and improved patient outcomes. This paper proposed a label-free and highly sensitive classification method for serum of patients with different degrees of gastric lesions by combining surface-enhanced Raman scattering (SERS) and machine learning analysis. Specifically, we prepared Au lotus-shaped (AuLS) nanoarrays substrates using seed-mediated and liquid-liquid interface self-assembly method for measuring SERS spectra of serum, and then the collected spectra were processed by principal component analysis (PCA) - multi-local means based nearest neighbor (MLMNN) model to achieve differentiation. By employing this pattern analysis, AuLS nanoarray substrates can achieve fast, sensitive, and label-free serum spectral detection. The classification accuracy can reach 97.5%, the sensitivity is higher than 96.7%, and the specificity is higher than 95.0%. Moreover, by analyzing the PCs loading plots, the most critical spectral features distinguishing different degrees of gastric lesions were successfully captured. This discovery lays the foundation for combining SERS with machine learning for real-time diagnosis and recognition of gastric lesions.

Eco-friendly second derivative synchronous fluorescence spectroscopic method for simultaneous determination of rosuvastatin and ezetimibe in pharmaceutical capsules

The fixed dose combination of Rosuvastatin and ezetimibe has recently received approval from the FDA for the treatment of elevated levels of low-density lipoprotein cholesterol in adults. Herein, an eco-friendly and highly sensitive spectrofluorimetric method was developed and validated for simultaneous determination of rosuvastatin and ezetimibe in commercial capsules. The developed method involved synchronous fluorescence spectroscopy combined with second derivative spectroscopy to resolve the overlapping fluorescence spectra of rosuvastatin and ezetimibe. The studied drugs were measured in synchronous mode at Δλ of 40 and their recorded synchronous fluorescence spectra were derivatized into second-order spectra, enabling the selective quantification of rosuvastatin and ezetimibe at 370 nm and 312 nm, respectively. Optimization studies regarding to the influence of buffer pH, incorporation of surfactant, choice of diluting solvent, and synchronous Δλ were carried out. The method was validated using the validation characteristics listed in ICH Q2(R1). The calibration curves displayed satisfactory linear relationships across the calibration range of 0.1–2 µg/mL for rosuvastatin and 0.05–3 µg/mL for ezetimibe. The methodology demonstrated robustness to minor modifications in the procedural parameters and selectivity in quantifying the studied drugs in synthetic mixed solutions and commercial capsules without interference. Furthermore, the level of environmental friendliness and sustainability of the suggested spectrofluorimetric approach was assessed in relation to two previously documented methodologies utilizing the AGREE metric. The findings indicated that the suggested method demonstrated a notably superior level of sustainability in comparison to the documented methods.

Multivariate optimization of the extraction and cleaning procedure to determine neonicotinoids in honey from intensive livestock and agricultural production areas by UHPLC-MS/MS

Neonicotinoid insecticides have been associated with the decline of bee population. This study aimed to investigate the occurrence of neonicotinoids in honey samples from apiaries located in areas of intensive livestock and agricultural production in Argentina. For this purpose, a QuEChERS-based analytical method was developed and optimized using multivariate strategies to reduce the processing time and the necessary amount of salts and sorbents. Validation was based on SANTE 11312/2021 guidelines, with recovery percentages between 79 and 120 %, and limits of quantitation between 0.25 and 1.00 µg kg−1. The present study provides a highly sensitive and reliable method for ascertaining residue levels of eight insecticides in 151 honey samples using liquid chromatography coupled to mass spectrometry (UHPLC-MS/MS). The presence of three of the target analytes was determined in 13 % of the samples, indicating that honey bee colonies were exposed to neonicotinoids, especially imidacloprid and thiamethoxam. The maximum residue limits were not exceeded in any of the samples; therefore, there is no risk to consumers from the public health point of view.

A dual-color ratiometric fluorescence sensor of biogenic amines with dye encapsulated covalent organic framework for meat freshness

Developing sensitive, nondestructive and real-time methods for assessing meat freshness is vital for ensuring food safety. Traditional monochromatic fluorescent probes for monitoring meat freshness markers like biogenic amines often face challenges in sensitivity, interference, visualization and portability for practical applications. Herein, we constructed a new Ru-Flu@COF fluorescence sensor, by combining a green fluorescent dye-encapsulated covalent organic framework (Flu@COF) with a red fluorescent compound (Ru(bpy)32+), exhibiting a dual inverse ratiometric fluorescence signal towards essential biogenic amines His and NH3·H2O, thus achieving a rich color gradient and spoilage-sensitive color discrimination. The detection linear range for His and NH3·H2O was 0.6∼200 μM and 0.5∼60 mg/L, with limits of detection of 0.5 μM and 0.2 mg/L, respectively. Furthermore, Ru-Flu@COF gel labels together with a smartphone enable portable, in situ and naked-eye monitoring freshness of pork, fish and shrimp, where the freshness level can be visually identified through the gel label color from red (fresh) to orange (less fresh), yellow (spoilage) and green (severe spoilage). The proposed smart label has the advantages of high sensitivity, excellent visual discrimination and portability, and it can be a promising platform for in situ nondestructive monitoring freshness of various kinds of meat for food safety inspection in the future.