Developed Method Research Articles

Portable and simultaneous detection of four respiratory pathogens through a microfluidic LAMP and real-time fluorescence assay.

Respiratory pathogen infections are seasonally prevalent and are likely to cause co-infections or serial infections during peak periods of infection. Since they often cause similar symptoms, simultaneous and on-site detection of respiratory pathogens is essential for accurate diagnosis and efficient treatment of these infectious diseases. However, molecular diagnostic techniques for multiple pathogens in this field are lacking. Herein, we developed a microfluidic LAMP and real-time fluorescence assay for rapid detection of multiple respiratory pathogens using a ten-channel microfluidic chip with pathogen primers pre-embedded in the chip reaction well. The microfluidic chip provided a closed reaction environment, effectively preventing aerosol contamination and improving the accuracy of the detection results. Its corresponding detection instrument could automatically collect and display the fluorescence curve in real time, which was more conducive to the interpretation of results. The results showed that the developed method could specifically recognize the nucleic acid of influenza A(H1N1), Mycoplasma pneumoniae, respiratory syncytial virus type A, and SARS-CoV-2 with low detection limits of 104 copies per mL or 103 copies per mL. The test results on clinical samples demonstrated that the developed method has high sensitivity (92.00%) and high specificity (100.00%) and even has the capability to differentiate mixed-infection samples. With simple operation and high detection efficiency, the present portable and simultaneous detection assay could significantly improve the efficiency of on-site detection of respiratory infectious diseases and promote the accurate treatment, efficient prevention and control of the diseases.

Development of a simple HPLC method for the determination of urinary O-aminohippuric acid (OAH) and an establishment of OAH reference interval

Background: O-aminohippuric acid (OAH) is considered a low-abundance urinary fluorescent metabolite with the potential to be an innovative lung cancer biomarker. There is a lack of simple methods for measuring urinary OAH metabolite, and the measurement needs to be normalized by urinary creatinine, which is produced at a constant rate. Thus, the newly developed method must be able to determine urinary creatinine and OAH simultaneously in the same run. Objective: This work aimed to develop and validate a simple high-performance liquid chromatography (HPLC) method for measuring urinary OAH and to establish the reference intervals of OAH in healthy individuals. Materials and methods: We synthesized OAH standard in a simple route and optimized simple HPLC method for simultaneous measurement of creatinine and OAH in a single run. Analysis was performed on a RP-C18 column with a gradient elution system of acetonitrile - ammonium acetate buffer (pH 6.5, 100 mM). After implementing optimal conditions, the procedure was compiled according to the International Council for Harmonisation (ICH) validation parameters. The developed method was used for the establishment of reference intervals of a total of 120 random urine samples of healthy individuals. Results: The linear range of the calibration curve for creatinine and OAH were 1-1000 µg/mL and 0.1-100 µg/mL, respectively. The recoveries ranged for both metabolites were between 91.35 % and 109.12%. The relative standard deviations (RSDs) for the intra-day and inter-day results ranged from 0.11-0.66 % to 0.16-1.73 %, respectively. The limits of detection (LOD) and quantification (LOQ) were 0.258 µg/mL and 0.783 µg/mL for creatinine, while OAH was 0.045 µg/mL and 0.137 µg/mL, respectively. The method was successfully applied to establish reference intervals of OAH in healthy individuals and was defined as 0.420-2.287 mmol/mol creatinine. Conclusion: According to various validated parameters, the proposed method was proven to quantify urinary OAH and creatinine in a single run. It can also be analyzed noninvasively without additional sample processing. Reported herein is the first establishment of OAH reference intervals in healthy individuals, which may benefit the utilization of OAH as a noninvasive biomarker for lung cancer detection in the future.

Analyzing the clinical efficacy and safety of levocetirizine based on its receptor occupancy, intraclass comparison and role in the treatment of CSU: an AROG consensus statement.

Chronic spontaneous urticaria (CSU) is characterized by urticaria persisting for more than 6 weeks and leading to significant morbidity. Antihistamines, especially sgAH (second generation antihistamines) are the first line of treatment for CSU. This consensus aimed to review the existing translational research on the receptor occupancy of antihistamines, including levocetirizine, and establish its role in the treatment of CSU. The consensus was led by the Antihistamine Receptor Occupancy Group (AROG) from India, an expert panel of twelve dermatologists from various regions with a mix of institutional and practitioner backgrounds. This consensus analyzed the existing translational research on the receptor occupancy of levocetirizine to establish its clinical efficacy and safety of levocetirizine in the treatment of CSU using the grading of recommendations assessment, development, and evaluation (GRADE) method as compared to the varied SGAH. Second-generation antihistamines constitute the first step in the therapeutic ladder for managing CSU. Levocetirizine has high bioavailability, a high affinity for and occupancy of the H1 receptor, a rapid onset of action, limited distribution, and minimal hepatic metabolism. It exhibits significant anti-inflammatory effects at clinically relevant concentrations. The marked receptor occupancy translates to better efficacy as compared to similarly dosed SGAH and the lower cost of the molecule makes it an appropriate drug for chronic use. Receptor occupancy should serve as the basis of intra-class head-to-head trials for CSU.

Мягкие навыки: концепции, проблемы, исследования

<p style="text-align: justify;"><span lang="EN-US">The work is aimed at clarifying the concept of “soft skills”. The materials of the study of concepts, problems, and research of soft skills in a professional environment are presented. The topic of soft skills is extremely relevant, both Russian and foreign students do not exactly understand what soft skills are, which may be explained by the fact that there is currently no single approach to understanding them. The purpose of studying this problem is to review concepts, problems and research on the topic of soft skills in foreign and national scientific literature. The results obtained make it possible to say that soft skills is a broad concept, and does not include a strictly defined list of qualities or abilities. There is evidence that researchers attribute a different set of abilities to soft skills, while different abilities will be most significant for different professional fields. Separately, such abilities can be distinguished as: communication skills, leadership position, decision-making skills, responsibility, empathy, self-esteem, critical thinking, analytical thinking, emotional intelligence (often assessed as part of soft skills in foreign studies). The most effective methods for developing soft skills include: methods for early development, educational tasks related to information search and analysis, critical thinking development, self-management (including the formulation of action plans), teamwork skills, and the presence of social and humanitarian disciplines (“Philosophy" and "Sociology”), as well as specialized courses such as “Team Building” and “Time Management”. It is also emphasized that the level of soft skills development among teachers is important for fostering soft skills in students.</span></p>

Squaraine-Linked Magnetic Covalent Organic Framework as a Solid-Phase Extraction Absorbent to Determine Trace Phenylpyrazoles.

Phenylpyrazoles are widely used pesticides in the food industry. It is highly desirable to develop efficient pre-treatment and analysis methods to extract and detect phenylpyrazoles in complex food matrices. Herein, the study reports novel squaraine-linked zwitterionic core-shell magnetic covalent organic frameworks (MCOFs), which are found to be excellent pretreatment materials for the detection of trace phenylpyrazoles in samples. By coupling MCOFs to magnetic solid-phase extraction (MSPE) with Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) analysis, the detection of phenylpyrazoles (fipronil, fipronil sulfone, fipronil sulfide, fipronil de-sulfoxide, fipronil desulfinyl, ethiprole, and flufiprole) is achieved and shows good linearity at concentrations of 1-800µg L-1 (R2 ≥ 0.9930). The limit of detection (LOD), limit of quantification (LOQ), and recovery rates are 0.01-0.50µg kg-1, 0.04-1.72µg kg-1, and 70.96-115.32%, respectively. More importantly, this method is successfully applied to determine the phenylpyrazoles in commercial egg, poultry, milk, jujube, cabbage, tea, and rice with a detection rate of ≈0.04%. Therefore, the developed method may contribute to a new strategy for the purification and multi-target extraction of complex food matrices.

Pembuatan Aplikasi Transaksi Pembayaran Makanan dan Minuman UMKM Nyonya Buki Kuningan

Food and beverage payment transaction applications are very important in helping MSMEs improve operational efficiency and service quality. This research aims to develop a web-based payment transaction application for Mrs. Buki Kuningan MSMEs. This application is designed using the Customer Relationship Management (CRM) method and tested with a qualitative approach. This research uses the Design and Development Research method. The stages in the design and development methods used are needs analysis, design, development, testing, and evaluation. The data collection techniques used include in-depth interviews, observations, and documentation. Data obtained from various sources will be analyzed using content analysis techniques, descriptive analysis, trial and evaluation. The test results show that this application can increase efficiency, reduce errors, and provide convenience for users. This application is expected to increase MSME income through improving service quality

Distributed learning for kernel mode–based regression

AbstractWe propose a parametric kernel mode–based regression built on the mode value, which provides robust and efficient estimators for datasets containing outliers or heavy‐tailed distributions. To address the challenges posed by massive datasets, we integrate this regression method with distributed statistical learning techniques, which greatly reduces the required amount of primary memory and simultaneously accommodates heterogeneity in the estimation process. By approximating the local kernel objective function with a least squares format, we are able to preserve compact statistics for each worker machine, facilitating the reconstruction of estimates for the entire dataset with minimal asymptotic approximation error. Additionally, we explore shrinkage estimation through local quadratic approximation, showcasing that the resulting estimator possesses the oracle property through an adaptive LASSO approach. The finite‐sample performance of the developed method is illustrated using simulations and real data analysis.

Detection, isolation and characterization of phage-host complexes using BONCAT and click chemistry

IntroductionPhages are viruses that infect prokaryotes and can shape microbial communities by lysis, thus offering applications in various fields. However, challenges exist in sampling, isolation and accurate prediction of the host specificity of phages as well as in the identification of newly replicated virions in response to environmental challenges.MethodsA new workflow using biorthogonal non-canonical amino acid tagging (BONCAT) and click chemistry (CC) allowed the combined analysis of phages and their hosts, the identification of newly replicated virions, and the specific tagging of phages with biotin for affinity chromatography.ResultsReplication of phage λ in Escherichia coli was selected as a model for workflow development. Specific labeling of phage λ proteins with the non-canonical amino acid 4-azido-L-homoalanine (AHA) during phage development in E. coli was confirmed by LC–MS/MS. Subsequent tagging of AHA with fluorescent dyes via CC allowed the visualization of phages adsorbed to the cell surface by fluorescence microscopy. Flow cytometry enabled the automated detection of these fluorescent phage-host complexes. Alternatively, AHA-labeled phages were tagged with biotin for purification by affinity chromatography. Despite biotinylation the tagged phages could be purified and were infectious after purification.DiscussionApplying this approach to environmental samples would enable host screening without cultivation. A flexible and powerful workflow for the detection and enrichment of phages and their hosts in pure cultures has been established. The developed method lays the groundwork for future workflows that could enable the isolation of phage-host complexes from diverse complex microbial communities using fluorescence-activated cell sorting or biotin purification. The ability to expand and customize the workflow through the growing range of compounds for CC offers the potential to develop a versatile toolbox in phage research. This work provides a starting point for these further studies by providing a comprehensive standard operating procedure.

Extended B‐spline‐based implicit material point method for saturated porous media

AbstractThe large deformation and fluidization process of a solid–fluid mixture includes significant changes to the temporal scale of the phenomena and the shape and properties of the mixed material. This paper presents an extended B‐spline (EBS)‐based implicit material point method (EBS‐MPM) for the coupled hydromechanical analysis of saturated porous media to enhance the overall versatility of MPM in addressing such diverse phenomena. The proposed method accurately represents phenomena such as high‐speed motion in both the quasi‐static and dynamic states by employing a full formulation of coupled hydromechanical modeling. The weak imposition of boundary conditions based on Nitsche's method allows representing the boundary conditions independent of the relative position of the particles and computational grid. In addition, it enables dynamic changes in the boundary domain based on the deformation. The robustness of this boundary representation is reinforced using EBS basis functions, which prevent the degradation of the condition number of the system matrices regardless of the position of the boundary domain with respect to the computational grid. Furthermore, a stabilization method based on a variational multiscale method (VMS) approach is employed to provide the flexibility in choosing arbitrary basis functions for spatial discretization, facilitating the effective construction of EBS. Numerical examples including comparisons between a full formulation and a simplified formulation are presented to demonstrate the performance of the developed method under various boundary conditions and loading states across different time scales.

Development and Validation of a Simple UV–HPLC Method to Quantify the Memantine Drug Used in Alzheimer’s Treatment

Memantine, a non-competitive NMDA receptor antagonist, is used to treat Alzheimer’s disease. Therefore, loading memantine in nanoparticles (NPs) could be an essential tool to improve the treatment effectiveness while reducing drug toxicity. Even though some approaches have been described to quantify memantine, none reported optimized methods using high-performance liquid chromatography resorting to ultraviolet detection (UV–HPLC) to determine encapsulation in NPs. The present research developed a HPLC method using pre-column derivatization for quantitatively analyzing memantine hydrochloride in NPs. Memantine was derivatized using 9-fluorenylmethyl chloroformate (FMOC). The developed method was fully validated regarding suitability, specificity, linearity, sensitivity, precision, accuracy, and robustness according to the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The retention time of memantine was 11.393 ± 0.003 min, with a mean recovery of 92.9 ± 3.7%. The new chromatographic method was validated and found to respond linearly over 5–140 μg/mL, with a high coefficient of determination. Intraday precision lay between 3.6% and 4.6%, and interday precision between 4.2% and 9.3%. The stability of memantine was also tested at 4 °C and −20 °C, and no signs of decay were found for up to 6 months. The new method was properly validated and proved simple, sensitive, specific, accurate, and precise for determining memantine encapsulation efficiency in lipid NPs. Greenness was evaluated, presenting a final score of 0.45. In the future, this methodology could also be applied to quantify memantine in different nanoformulations.

Pengaruh Penggunaan Media Assembler Terhadap Motivasi dan Prestasi Belajar Siswa pada Materi Sistem Pernapasan

Media is a form of communication that aims to send messages from the sender to the recipient in oral or written form. According to Sriyanti, the types of media that are often used in learning include; print media, practical media, interactive media, E-Learning and M-Learning. The aim of the research carried out was to determine the effect of using assembler media on student motivation and learning achievement in respiratory material. This research used research and development methods. This is a research method used for a particular product and testing the effectiveness of the product. The increase in students' conceptual understanding reflects the positive influence of Assembler media on student learning achievement. Apart from that, the use of this media also has the potential to increase students' learning motivation, although direct evaluation of motivation was not carried out in this research. However, positive responses from students to such media can be considered as a potential indicator of increased motivation

Pengembangan Media Pembelajaran Multimedia Interaktif pada Materi Ekologi dan Keanekaragaman Hayati Indonesia untuk Peserta Didik Kelas VII di SMP Negeri 2 Darma

The purpose of this research is to develop and determine the feasibility of interactive multimedia content in science class VII of Indonesia's ecology and biodiversity material. This research uses a Research and Development (R&D) method with a development model called ADDIE. This development model has five phases, namely analysis, design, development, implementation, and evaluation. The sample in this study was taken from 1 (one) class at SMP Negeri 2 Darma which amounted to 25 students. The data collection techniques in this study are observation, interviews, and questionnaires. The end result of this research is the development of products in the form of interactive multimedia content. The development of interactive multimedia was tested through a questionnaire for validation by media experts and obtained a feasibility level of 85% which is included in the "Very Feasible" feasibility level, and material experts realized that they got a feasibility percentage of 92% including the "Very Feasible" category. A small group experiment involving five students of SMP Negeri 2 Darma produced a result of 93% including the "Very Feasible" category. The results of the large group test with a total of 20 students showed a feasibility of 93% including the "Very Feasible" category. The average percentage of results from media experts, material experts, small-scale and large-scale trials got 91% with the category of "Very Feasible"

Palladium-Catalyzed Cross-Coupling of Aryl Bromides and Chlorides with Trimethylsilylalkynes under Mild Conditions.

Herein, we disclose a palladium-catalyzed cross-coupling of aryl bromides and chlorides with trimethylsilylalkynes under mild reaction conditions. This method utilizes commercially available and air stable palladium precatalysts and avoids the use of copper cocatalysts. Moreover, it allows for the synthesis of a wide range of disubstituted alkynes in high yields with excellent functional group tolerance. The utility of the developed method was further demonstrated via the late-stage alkynylation of pharmaceuticals and natural bioactive compounds.

Development of an Internet of Things-based Environmental Information System for Realtime Monitoring of Air Conditions

This study aims to develop an environmental information system called the Environmental Monitoring System (EENNOS) to monitor air environmental conditions. ENNOS is an Internet of Things (IoT) based system that monitors air environmental conditions in the form of temperature, humidity and carbon dioxide. The research was conducted using the 4D development method which consisted of the Define, Design, Develop, and Disseminate stages. EENNOS was developed using a DHT 22 sensor for DHT 22 temperature and humidity and a carbon dioxide sensor with MQ-135. The controller used is Arduino uno ESP8266 with output sending using the internet to the database and presented on the dashboard and displayed on a 16x2 liquid crystal screen. The output is in the form of dashboard innovations that are registered with copyrights and scientific publications in the form of articles in the Journal of Environmental Sciences.

Hybrid Visual Odometry Algorithm Using a Downward-Facing Monocular Camera

The increasing interest in developing robots capable of navigating autonomously has led to the necessity of developing robust methods that enable these robots to operate in challenging and dynamic environments. Visual odometry (VO) has emerged in this context as a key technique, offering the possibility of estimating the position of a robot using sequences of onboard cameras. In this paper, a VO algorithm is proposed that achieves sub-pixel precision by combining optical flow and direct methods. This approach uses only a downward-facing, monocular camera, eliminating the need for additional sensors. The experimental results demonstrate the robustness of the developed method across various surfaces, achieving minimal drift errors in calculation.

Penerapan Aplikasi Manajemen Laboratorium Dinas Lingkungan Hidup Kota Palembang

Waste samples that is managed properly will generate added value and provide benefits for environmental management. The purpose of this study is to design a web-based waste water sample data management information system so that it can make it easier for waste laboratory staff and the public as customers and admins to manage and access waste water sample data and transactions. The method used in data collection is the DevOps method and it is also not important as a researcher to interview officials from the Palembang city environmental service regarding the laboratory management application program.. In making the website, the author uses the PHP and MySQL programming languages with the DevOps development method, object-based software modeling using UML and designing the system interface to show system functions. Database design is made to show the relationship between entities and data contained in the system. Users who will use the waste laboratory management application information system are employees of the environmental service laboratory unit as admin and the public. The results of this study indicate that the website design that was built can make it easier for the admin to manage waste water sample data, it can be stored in a database and can be accessed at any time. Apart from that, for residents,, this system has the benefit of accelerating the process of collecting waste in their environment, this system has the benefit of speeding up the process of collecting waste water sample data.

Analysis of phthalate esters from vegetable oils by gas chromatography-mass spectrometry coupled with headspace solid-phase microextraction using cyclodextrin-based hypercrosslinked polymer coated fiber

Phthalate esters (PAEs) are used as additives to enhance the pliability and malleability of plastics. These substances frequently migrate from packaging materials to vegetable oils because of the absence of covalent bonds. Over time, this migration could result in the accumulation of PAEs in the human body through ingestion, contributing to various diseases. Therefore, accurate qualitative and quantitative analyses of PAEs in vegetable oils are imperative to assess the origins of contamination and investigate their toxicity, degradation, migration, and transformation patterns. However, the concentration of PAEs in most samples is low, and the composition of vegetable oils is complex. Thus, PAEs must be enriched and purified using appropriate sample pretreatment procedures before analysis. Common methods for pretreating PAEs in oil include solid-phase extraction (SPE), dispersive SPE, and magnetic SPE. These techniques require time-consuming and labor-intensive procedures such as oil dissolution, solvent extraction, and degreasing. These approaches also require numerous solvents and containers, increasing the risk of sample cross-contamination. Solid-phase microextraction (SPME) integrates sampling, extraction, purification, concentration, and injection into a single process, significantly accelerating analytical testing and reducing the potential for sample cross-contamination. In headspace (HS) mode, the analytes achieve equilibrium on the coating and are extracted in the gas phase. The fibers are shielded from nonvolatile and high-relative molecular mass substances in the sample matrix. Thus, SPME is an ideal method for extracting volatile compounds in vegetable oils. When HS-SPME coupled with gas chromatography-mass spectrometry (GC-MS), it can achieve the rapid screening of PAEs in vegetable oil. In this study, an SPME with cyclodextrin-based hypercrosslinked polymers (BnCD-HCP) coated on stainless steel fibers was employed to extract PAEs from vegetable oil. The structure and morphology of the polymers were characterized using Fourier-transform infrared spectroscopy, nuclear magnetic spectroscopy, and scanning electron microscopy. BnCD-HCP exhibited high stability and diverse interactions, including π-π, hydrophobic, and host-guest interactions. The oil samples were incubated with methanol, and the PAEs were extracted from the headspace using the probe. The optimal extraction parameters included an extraction time of 20 min, extraction temperature of 50 ℃, desorption time of 4 min, and desorption temperature of 275 ℃. The BnCD-HCP/HS-SPME method was evaluated under optimized experimental conditions. The limits of detection (LODs) and quantification (LOQs) were determined by applying signal-to-noise ratios (S/N) of 3 and 10, respectively. Method accuracy was evaluated using relative standard deviations (RSDs). Single-needle precision was evaluated by conducting three consecutive analyses at 3 h intervals within a day. Inter-needle precision was assessed by conducting the same analyses (three replicates) with differently coated fibers. The 12 PAE compounds exhibited good linearity with correlation coefficients (R2) of at least 0.99. The LODs and LOQs ranged from 0.21 to 3.74 μg/kg and from 0.69 to 12.34 μg/kg, respectively. The RSDs were in the range of 1.8%-11.4% and 5.1%-13.9% for the single-needle and needle-to-needle methods, respectively. The proposed method was applied to soybean, peanut, and sunflower oils, and two PAEs were found in all three oils. Moreover, the method demonstrated good precision (RSD=1.17%-11.73%) and recoveries (72.49%-124.43%). Compared with other methods, the developed method was able to extract many target analytes and had a low or comparable LOD and high recovery. More importantly, this method does not require tedious operations such as solvent extraction and purification. Consequently, the developed method can be used to extract not only PAEs in oils but also other substances with a high lipid content.

Development of a 2A peptide-based multigene expression system and its application for enhanced production of ganoderic acids in Ganoderma lucidum

Ganoderma has received much attention for its medicinal value, but the manipulation of multiple genes remains a challenge, hindering the genetic engineering of this species for the development of cell factories. Here, we first showed that the presence of an intron is necessary for the efficient expression of the endogenous cDNA of carboxin-resistant gene (cbx) in G. lucidum. Then, the self-cleaving function of 2A peptide was investigated in G. lucidum by linking cbx cDNA to the codon-optimized hygromycin B-resistant gene (ophph) using the 2A-peptide sequence. The results showed that cbx cDNA and ophph can be successfully expressed in G. lucidum in a bicistronic manner from a single transcript. Moreover, the expression of both genes was not affected by the order within the 2A cassette. In addition, simultaneous expression of cbx cDNA, ophph, and codon-optimized yellow fluorescent protein gene (opyfp) was conducted for the first time in G. lucidum using the 2A peptide-based approach. The developed method was successfully applied to express both cDNA of the 3-hydroxy-3-methylglutaryl coenzyme A reductase (hmgr) and squalene epoxidase gene (se) for enhanced production of ganoderic acids (GAs) in G. lucidum. The engineered strain produced the maximum content of GA-Mk, GA-T, GA-S, and GA-Me were 26.56±3.53,39.58±3.75, 16.54±2.16, and 19.1±1.87 μg/100mg dry weight, respectively. These values were 3.85-, 4.74-, 3.65-, and 3.23-fold higher than those produced by the control strain. The developed method will be useful for the manipulation of complex metabolic or regulatory pathways involving multiple genes in Ganoderma.

Determination of eight neonicotinoid pesticides in wastewater by solid phase extraction combined with liquid chromatography-tandem mass spectrometry

Neonicotinoid pesticides are a relatively new class of pesticides that have garnered significant attention owing to their potential ecological risks to nontarget organisms. A method combining solid phase extraction with liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) was developed for the rapid and accurate detection of eight neonicotinoid pesticides (dinotefuran, E-nitenpyram, thiamethoxam, clothianidin, imidacloprid, imidaclothiz, acetamiprid, and thiacloprid) in wastewater. The chromatographic mobile phase and MS parameters were selected, and a single-factor method was used to determine the optimal column type, extraction volume, sample loading speed, and pH for SPE. The optimal parameters were as follows: column type, HLB column (500 mg/6 mL); sample extraction volume, 500 mL; sample loading speed, 10 mL/min; and sample pH, 6-8. The matrix effects of the wastewater samples were reduced by optimizing the chromatographic gradient-elution program, examining the dilution factor of the samples, and using the isotope internal standard calibration method. Prior to analysis, the wastewater samples were diluted 5-fold with ultrapure water for pretreatment. Subsequently, 2 mmol/L ammonium acetate aqueous solution containing 0.1% (v/v) formic acid and methanol was used as mobile phases for gradient elution on a ZORBAX Eclipse Plus C18 column (100 mm×2.1 mm, 1.8 μm). The samples were quantified using positive-ion multiple reaction monitoring (MRM) mode for 10 min. Imidacloprid-d4 was used as the isotope internal standard. The SPE process was further optimized by applying response surface methodology to select the type and mass of rinsing and elution solvents. The optimal pretreatment of the SPE column included rinsing with 10% methanol aqueous solution and elution with methanol-acetonitrile (1∶1, v/v) mixture (7 mL). The eight neonicotinoid pesticides showed satisfactory linearity within the relevant range, with linear correlation coefficients (r) all greater than 0.9990. The method detection limits (MDLs) ranged from 0.2 to 1.2 ng/L, and the method quantification limits (MQLs) ranged from 0.8 to 4.8 ng/L. The average recoveries of the eight neonicotinoid pesticides were in the range of 82.6%-94.2% at three spiked levels, with relative standard deviations (RSDs) ranging from 3.9% to 9.4%. Finally, the optimized method was successfully applied to analyze wastewater samples collected from four sewage treatment plants. The results indicated that the eight neonicotinoid pesticides could be generally detected at concentrations ranging from not detected (ND) to 256 ng/L. The developed method has a low MDL and high accuracy, rendering it a suitable choice for the trace detection of the eight neonicotinoid pesticides in wastewater when compared with other similar methods. The proposed method can be utilized to monitor the environmental impact and assess the potential risks of neonicotinoid pesticides in wastewater, thus promoting the protection of nontarget organisms and the sustainable use of these pesticides in agriculture.

Separation of triacylglycerol (TAG) isomers by cyclic ion mobility mass spectrometry

Triacylglycerols (TAGs), a major lipid class in foods and the human body, consist of three fatty acids esterified to a glycerol backbone. They can occur in various isomeric forms, including sn-positional, cis/trans configurational, acyl chain length, double bond positional, and mixed type isomers. Separating isomeric mixtures is of great interest as different isomers can have distinct influence on mechanisms, such as digestibility, oxidative stability, or lipid metabolism. However, TAG isomer separation remains challenging with established analytical methodologies such as liquid-chromatography coupled to mass spectrometry (LC-MS). In this study, we developed a method with cyclic ion mobility mass spectrometry (cIMS-MS) for the separation and identification of all types of TAG isomers. First, the influence of different adducts (Li+, NH4+, Na+, and K+) on the separation was studied. Overall, it was concluded that the sodium adduct is the best choice to efficiently separate all types of TAG isomers. In addition, trends were found in the influence of specific structural features on the drift time order. An order of relative influence (from high to low) was established; (1) degree of unsaturation of the fatty acid(s) on an exterior position (if the total degree of unsaturation(s) is equal in both TAGs), (2) acyl chain length on the exterior positions, (3) cis/trans configuration, and (4) double bond (DB)-position. Finally, various cIMS-MS strategies were developed for the separation of mixtures containing four, five, and six isomers. To conclude, the developed methods can be used for separation of complex mixtures of TAG isomers and have great potential to be expanded to isomers of similar types of lipids such as di- and monoacylglycerols. This study also shows the potential of cIMS-MS to be used for the application on real TAG samples.