Development of a rapid screening method utilizing 2D LC for effect-directed analysis in the identification of environmental toxicants.

Effect-directed analysis (EDA) is a crucial tool in environmental toxicology, effectively integrating toxicity testing with chemical analysis. The conventional EDA approach, however, presents challenges such as significant solvent consumption, extended analysis time, labor intensity, and potential contamination risks. In response, we introduce an innovative alternative to the conventional EDA. This method utilizes the MTT bioassay and online two-dimensional liquid chromatography (2D LC) coupled with high-resolution mass spectrometry (HR-MS), significantly reducing the fractionation steps and leveraging the enhanced sensitivity of the bioassay and automated chemical analysis. In the chemical analysis phase, a switching valve interface is employed for comprehensive analysis. We tested the performance of both the conventional and our online 2D LC-based methods using a household product. Both methods identified the same number of toxicants in the sample. Our alternative EDA is 22.5 times faster than the conventional method, fully automated, and substantially reduces solvent consumption. This novel approach offers ease, cost-effectiveness, and represents a paradigm shift in EDA methodologies. By integrating a sensitive bioassay with online 2D LC, it not only enhances efficiency but also addresses the challenges associated with traditional methods, marking a significant advancement in environmental toxicology research.

Payload Identification and Gravity/Inertial Compensation for Six-Dimensional Force/Torque Sensor with a Fast and Robust Trajectory Design Approach.

In the robot contact operation, the robot relies on the multi-dimensional force/torque sensor installed at the end to sense the external contact force. When the effective load and speed of the robot are large, the gravity/inertial force generated by it will have a non-negligible impact on the output of the force sensor, which will seriously affect the accuracy and effect of the force control. The existing identification algorithm time is often longer, which also affects the efficiency of force control operations. In this paper, a self-developed multi-dimensional force sensor with integrated gravity/inertial force sensing function is used to directly measure the resultant force. Further, a method for the rapid identification of payload based on excitation trajectory is proposed. Firstly, both a gravity compensation algorithm and an inertial force compensation algorithm are introduced. Secondly, the optimal spatial recognition pose based on the excitation trajectory was designed, and the excitation trajectory of each joint is represented by a finite Fourier series. The least square method is used to calculate the identification parameters of the load, the gravity, and inertial force. Finally, the experiment was verified on the robot. The experimental results show that the algorithm can quickly identify the payload, and it is faster and more accurate than other algorithms.

Electrochemical Fingerprint Biosensor for Natural Indigo Dye Yielding Plants Analysis.

Indigo is a plant dye that has been used as an important dye by various ancient civilizations throughout history. Today, due to environmental and health concerns, plant indigo is re-entering the market. Strobilanthes cusia (Nees) Kuntze is the most widely used species in China for indigo preparation. However, other species under Strobilanthes have a similar feature. In this work, 12 Strobilanthes spp. were analyzed using electrochemical fingerprinting technology. Depending on their electrochemically active molecules, they can be quickly identified by fingerprinting. In addition, the fingerprint obtained under different conditions can be used to produce scattered patter and heatmap. These patterns make plant identification more convenient. Since the electrochemically active components in plants reflect the differences at the gene level to some extent, the obtained electrochemical fingerprints are further used for the discussion of phylogenetics.

Pharmacist-Driven Implementation of Fast Identification and Antimicrobial Susceptibility Testing Improves Outcomes for Patients with Gram-Negative Bacteremia and Candidemia.

Bloodstream infections (BSI) are associated with increased morbidity and mortality, especially when caused by Gram-negative or fungal pathogens. The objective of this study was to assess the impact of fast identification-antimicrobial susceptibility testing (ID/AST) with the Accelerate Pheno system (AXDX) from May 2018 to December 2018 on antibiotic therapy and patient outcomes. A pre-post quasiexperimental study of 200 patients (100 pre-AXDX implementation and 100 post-AXDX implementation) was conducted. The primary endpoints measured were time to first antibiotic intervention, time to most targeted antibiotic therapy, and 14-day hospital mortality. Secondary endpoints included hospital and intensive care unit (ICU) length of stay (LOS), antibiotic intensity score at 96 h, and 30-day readmission rates. Of 100 patients with Gram-negative bacteremia or candidemia in each cohort, 84 in the preimplementation group and 89 in the AXDX group met all inclusion criteria. The AXDX group had a decreased time to first antibiotic intervention (26.3 versus 8.0, P = 0.003), hours to most targeted therapy (14.4 versus 9, P = 0.03), hospital LOS (6 versus 8, P = 0.002), and average antibiotic intensity score at 96 h (16 versus 12, P = 0.002). Both groups had a comparable 14-day mortality (0% versus 3.6%, P = 0.11). In this analysis of patients with Gram-negative bacteremia or candidemia, fast ID/AST implementation was associated with decreased hospital LOS, decreased use of broad-spectrum antibiotics, shortened time to targeted therapy, and an improved utilization of antibiotics within the first 96 h of therapy.

Fast Identification of Cyathula officinalis and Its Adulterant and Doped Adulterant Based on Properties and Foam Test / 中国实验方剂学杂志

Objective: To rapidly identify Cyathula officinalis and its adulterant C. capitata and C. officinalis*C. capitata and doped adulterant. Method: Properties combined with foam test method were used for identifying C.officinalis, its adulterant and doped adulterant. In the aspect of properties,6 aspects including shape,size,texture,color,smell and taste,were observed, smelled and tasted. In the aspect of foam test,the volume of foam produced was used as the determination index to investigate the sample amount,water amount, shaking time,particle size,water temperature,repeatability,adulteration ratio and its stability. Result: In the aspect of properties,C. officinalis and its adulterant showed obvious difference in the shape,size,color,texture,smell and taste,especially the red color and bitter taste of its adulterant. In the aspect of foam test,the optimum parameters were as follows:sample particle (screened with 3 sieves) 0.3 g,a test tube with plug and scale,water 10 mL and airtight,forced shaking up and down for 1 min,settling for 5 min. Such method can be used to identify C. officinalis, its adulterant and doped adulterant. The volume of foams produced by C. officinalis and its adulterant and different ratio of doped adulterant showed no change within 5-30 min,slightly decreases after 9 h; the higher adulteration ratio; the larger volume of foam and better stability. The 8 batches of C. officinalis and 8 batch of adulterants proved that the volume of the foams produced was all less than 2 mL in the C. officinalis,more than 13 mL in the adulterant is,and more than 5 mL in 5% doped adulterant, showing statistical difference. From the properties combined with foam test,5 specific identification elements were obtained for identifying C. officinalis, its adulterant and doped adulterant. Conclusion: Through the 5 specific identification elements,the properties combined with foam test can be used to distinguish the C. officinalis from its adulterant C. officinalis and C. officinalis*C. capitata and doped adulterant,characterized by accuracy,simpleness,short time,low cost and feasibility. It can provide a new method and reference for identifying C.officinalis from its adulterant and doped adulterant.

Dynamic surface-enhanced Raman spectroscopy and Chemometric methods for fast detection and intelligent identification of methamphetamine and 3, 4-Methylenedioxy methamphetamine in human urine.

Conventional Surface-Enhanced Raman Spectroscopy (SERS) for fast detection of drugs in urine on the portable Raman spectrometer remains challenges because of low sensitivity and unreliable Raman signal, and spectra process with manual intervention. Here, we develop a novel detection method of drugs in urine using chemometric methods and dynamic SERS (D-SERS) with mPEG-SH coated gold nanorods (GNRs). D-SERS combined with the uniform GNRs can obtain giant enhancement, and the signal is also of high reproducibility. On the basis of the above advantages, we obtained the spectra of urine, urine with methamphetamine (MAMP), urine with 3, 4-Methylenedioxy Methamphetamine (MDMA) using D-SERS. Simultaneously, some chemometric methods were introduced for the intelligent and automatic analysis of spectra. Firstly, the spectra at the critical state were selected through using K-means. Then, the spectra were proposed by random forest (RF) with feature selection and principal component analysis (PCA) to develop the recognition model. And the identification accuracy of model were 100%, 98.7% and 96.7%, respectively. To validate the effect in practical issue further, the drug abusers'urine samples with 0.4, 3, 30ppm MAMP were detected using D-SERS and identified by the classification model. The high recognition accuracy of >92.0% can meet the demand of practical application. Additionally, the parameter optimization of RF classification model was simple. Compared with the general laboratory method, the detection process of urine's spectra using D-SERS only need 2 mins and 2µL samples volume, and the identification of spectra based on chemometric methods can be finish in seconds. It is verified that the proposed approach can provide the accurate, convenient and rapid detection of drugs in urine.

Identification of unclassified influenza A virus using high-throughput sequencing technology / 中华实验和临床病毒学杂志

Objective@#To identify the avian influenza virus subtype from the avian and environmental samples using the Ion Torrent new-generation semiconductor sequencing technology and to establish a high-throughput sequencing method to identify unclassified influenza A virus.@*Methods@#Virus RNA was extracted from the nine avian swab and environmental samples and real-time RT-PCR was carried out to detect universal fluA, H5N1, H7N9 and H9N2. The whole genome of influenza A virus was amplified by PathAmpFluA kit. Sequencing library was prepared using Next Fast DNA Fragmentation & Library Prep Set for Ion Torrent kit and high-throughput sequencing was done by Ion Torrent Personal Genome Machine(PGM). Data from the PGM was processed and quality evaluated using Ion TorrentSuite v3.0 software. Sequence assembly and influenza database blast were carried out by FluAtyping v4.0 and PathogenAnalyzer bioinformatics software to identify the influenza A virus subtype of these nine samples.@*Results@#The results of real-time RT-PCR for universal fluA of these nine samples were positive but the results for H5N1, H7N9 and H9N2 were negative. Seven subtypes of influenza A virus were identified by high-throughput sequencing and bioinformatics analysis: six samples were H2N3, H5N6, H5N8, H7N1, H7N7, H11N3 subtype respectively and three samples were H6N6 subtype.@*Conclusions@#Avian influenza virus has many subtypes in the environment of Zhejiang province. Ion Torrent semiconductor sequencing technology is suitable for fast identification of unclassified influenza virus for avian influenza environment monitoring.

Short-term incubation of positive blood cultures in brain-heart infusion broth accelerates identification of bacteria by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry.

PURPOSE: Fast identification of bacteria directly from positive blood cultures (BCs) by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS) can be achieved either using the MALDI Sepsityper kit (protein extraction method) or after a short-term pre-cultivation step on solid medium. We developed a new method that involves short-term enrichment of positive BCs in brain-heart infusion broth (BHI) prior to MALDI-TOF MS analysis. METHODOLOGY: Eighty-four BCs flagged as positive were included in this study; these were processed in parallel either directly using the MALDI Sepsityper kit or following a short-term culture either in BHI or on Columbia blood agar with 5 % sheep blood (CBA). RESULTS: Bacterial species were successfully identified in 91.6, 89.2 and 65.4 % of cases after pre-cultivation for 4 h in BHI, on CBA, or by using the MALDI Sepsityper kit, respectively. Overall, the mean incubation time to correct identification was shorter when pre-cultures were performed in BHI; the mean time for Gram-negative rods was 78.2 min in BHI and 108.2 min on CBA (P=0.045), and the mean time for Gram-positive cocci was 128.5 min in BHI and 169.6 min on CBA (P=0.013). CONCLUSION: Short-term enrichment of BCs in BHI accelerates identification of a number of bacterial species by MALDI-TOF MS. Further prospective studies are needed to validate our method and gauge its potential clinical impact on the management of bloodstream bacterial infections.

[Fast identification of constituents of Lagotis brevituba by using UPLC-Q-TOF-MS/MS method].

The chemical constituents of Lagotis brevituba were rapidly determined and analyzed by using ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) method, providing material basis for the clinical application of L. brevituba. The separation was performed on UPLC YMC-Triart C18 (2.1 mm×100 mm, 1.9 µm) column, with acetonitrile-water containing 0.2% formic acid as mobile phase for gradient elution. The flow rate was 0.4 mL•min-1 gradient elution and column temperature was 40 ℃, the injection volume was 2 µL. ESI ion source was used to ensure the data collected in a negative ion mode. The chemical components of L. brevituba were identified through retention time, exact relative molecular mass, cleavage fragments of MS/MS and reported data. The results showed that a total of 22 compounds were identified, including 11 flavones, 6 phenylethanoid glycosides, 1 iridoid glucosides, and 4 organic acid. The UPLC-Q-TOF-MS/MS method could fast identify the chemical components of L. brevituba, providing valuable information about L. brevituba for its clinical application.

Fast identification of constituents of Lagotis brevituba by using UPLC-Q-TOF-MS/MS method / 中国中药杂志

The chemical constituents of Lagotis brevituba were rapidly determined and analyzed by using ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) method, providing material basis for the clinical application of L. brevituba. The separation was performed on UPLC YMC-Triart C₁₈ (2.1 mm×100 mm, 1.9 μm) column, with acetonitrile-water containing 0.2% formic acid as mobile phase for gradient elution. The flow rate was 0.4 mL•min-1 gradient elution and column temperature was 40 ℃, the injection volume was 2 μL. ESI ion source was used to ensure the data collected in a negative ion mode. The chemical components of L. brevituba were identified through retention time, exact relative molecular mass, cleavage fragments of MS/MS and reported data. The results showed that a total of 22 compounds were identified, including 11 flavones, 6 phenylethanoid glycosides, 1 iridoid glucosides, and 4 organic acid. The UPLC-Q-TOF-MS/MS method could fast identify the chemical components of L. brevituba, providing valuable information about L. brevituba for its clinical application.