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rapid and sensitive profiling and quantification of the human cell line proteome by LC-MS/MS without prefractionation

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PROTEOMICS 2014年第17-18期14卷 2008-2016页

作者： Yin, Xuefei Liu, Xiaohui Zhang, Yang Yan, Guoquan Wang, Fang Lu, Haojie Shen, Huali Yang, Pengyuan Fudan Univ Shanghai Med Sch Dept Chem Shanghai 200032 Peoples R China Fudan Univ Shanghai Med Sch Inst Biomed Sci Shanghai 200032 Peoples R China

In this paper, we demonstrate a rapid and reproducible 1D LC-MS/MS workflow for fast quantitative proteomic research. We have optimized the LC-MS/MS conditions, including digestion and gradient conditions, sample loading amount, and MS parameter settings. As a result, we were able to obtain twice as many protein identifications compared with the LC-MS/MS conditions before optimization. More than 4500 protein groups and 50 000 peptides were identified in less than 8 h without any fractionation. This 1D workflow was then applied to the analysis of the MLN4924 treated/untreated human umbilical vein endothelial cell (HUVEC) samples with label-free quantification. In these experiments, a total of 179 proteins showed a statistically significant expression change after the MLN4924 treatment. Functional analysis showed that these proteins are associated with cell death and survival;gene expression;cell cycle;and DNA replication, recombination, and repair.

关键词： 1D LC-MS/MS High throughput rapid quantification Technology

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A Green and rapid High-Performance Liquid Chromatography-Ultraviolet Method for Determination of Four Organic Acids in Different Parts of Dandelion With Blue Applicability Grade Index Assessment

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JOURNAL OF FOOD BIOCHEMISTRY 2025年第1期2025卷

作者： Chen, Jing Lei, Qinggui Yang, Weiqi Wang, Wenhao Tan, Guang' e Qian, Zhengming Xiangnan Univ Affiliated Hosp Dept Rehabil Med Chenzhou 423000 Peoples R China Xiangnan Univ Coll Med Imaging Lab & Rehabil Chenzhou 423000 Peoples R China Dongguan HEC Cordyceps R&D Co Ltd Key Lab State Adm Tradit Chinese Med Dongguan 523871 Peoples R China Jinan Univ Inst Tradit Chinese Med & Nat Prod Coll Pharm Guangzhou 510632 Peoples R China

Dandelion is a functional food that serves both medicinal and dietary purposes, which requires quality control to ensure its safety and efficacy. This study established a rapid high-performance liquid chromatography-ultraviolet (HPLC-UV) method for analyzing four organic acids in different parts of dandelion. After ultrasonic extraction with 40% methanol, samples were analyzed using an Agilent InfinityLab Poroshell 120 EC-C-18 column with a mobile phase of 0.1% formic acid aqueous-methanol and detection wavelength at 325 nm. The method completed the analysis in 5 min (2 min for extraction and 3 min for chromatographic analysis). The 4 organic acid standards (caftaric acid, chlorogenic acid, caffeic acid, and chicoric acid) showed excellent linearity (r > 0.9995), and the average spiked recoveries were 101.38%, 96.85%, 103.94%, and 100.15% (relative standard deviations were 1.00%, 1.00%, 0.97%, and 0.55%), respectively. The total content of 4 organic acids is highest in dandelion flowers (1.355 mg/g dry weight [DW]), followed by stems/leaves (1.182 mg/g DW), and lowest in roots (0.026 mg/g DW). Additionally, to evaluate the environmental impact of both new and reported methods, this study performed an analysis using the blue applicability grade index. The developed methodology obtained the highest score of 80.0, which indicated that the developed HPLC method was most effective and eco-friendly. In conclusion, this method is rapid, accurate, and eco-friendly, which is help to the product development and quality evaluation of dandelion.

关键词： dandelion environmental sustainability functional food green chemistry organic acids quality evaluation rapid quantification

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Novel method for rapid, simultaneous T₁, T₂^*, and proton density quantification

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MAGNETIC RESONANCE IN MEDICINE 2007年第3期57卷 528-537页

作者： Warntjes, J. B. M. Dahlqvist, O. Lundberg, P. Linkoping Univ Ctr Med Imaging & Visualizat SE-58185 Linkoping Sweden Linkoping Univ Dept Med & Care Div Clin Physiol SE-58185 Linkoping Sweden Linkoping Univ Dept Med & Care Div Radiat Phys SE-58185 Linkoping Sweden Linkoping Univ Hosp Dept Radiat Phys S-58185 Linkoping Sweden

An imaging method called "quantification of relaxation times and proton density by twin-echo saturation-recovery turbo-field echo" (ORAPTEST) is presented as a means of quickly determining the longitudinal T-1 and transverse T-2* relaxation time and proton density (PD) within a single sequence. The method also includes an estimation of the B, field inhomogeneity. High-resolution images covering large volumes can be achieved within clinically acceptable times of 5-10 min. The range of accuracy for determining T-1, T-2*, and PD values is flexible and can be optimized relative to any anticipated values. We validated the experimental results against existing methods, and provide a clinical example in which quantification of the whole brain using 1.5 mm(3) voxels was achieved in less than 8 min.

关键词： T-1 mapping T-2(*) mapping proton density mapping quantitative MRI rapid quantification

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Truncated M13 phage for smart detection of E. coli under dark field

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JOURNAL OF NANOBIOTECHNOLOGY 2024年第1期22卷 599页

作者： Yuan, Jiasheng Zhu, Huquan Li, Shixinyi Thierry, Benjamin Yang, Chih-Tsung Zhang, Chen Zhou, Xin Yangzhou Univ Inst Comparat Med Coll Vet Med Yangzhou 225009 Peoples R China Yangzhou Univ Jiangsu Coinnovat Ctr Prevent & Control Important Yangzhou 225009 Peoples R China Yangzhou Univ Joint Int Res Lab Agr & Agriprod Safety Minist Educ China Yangzhou 225009 Peoples R China Univ South Australia Future Ind Inst Mawson Lakes Campus Adelaide SA 5095 Australia Yangzhou Univ Sch Informat Engn Yangzhou 225127 Peoples R China Yangzhou Univ Jiangsu Prov Engn Res Ctr Knowledge Management & I Yangzhou 225127 Peoples R China

Background The urgent need for affordable and rapid detection methodologies for foodborne pathogens, particularly Escherichia coli (E. coli), highlights the importance of developing efficient and widely accessible diagnostic systems. Dark field microscopy, although effective, requires specific isolation of the target bacteria which can be hindered by the high cost of producing specialized antibodies. Alternatively, M13 bacteriophage, which naturally targets E. coli, offers a cost-efficient option with well-established techniques for its display and modification. Nevertheless, its filamentous structure with a large length-diameter ratio contributes to nonspecific binding and low separation efficiency, posing significant challenges. Consequently, refining M13 phage methodologies and their integration with advanced microscopy techniques stands as a critical pathway to improve detection specificity and efficiency in food safety diagnostics. Methods We employed a dual-plasmid strategy to generate a truncated M13 phage (tM13). This engineered tM13 incorporates two key genetic modifications: a partial mutation at the N-terminus of pIII and biotinylation at the hydrophobic end of pVIII. These alterations enable efficient attachment of tM13 to diverse E. coli strains, facilitating rapid magnetic separation. For detection, we additionally implemented a convolutional neural network (CNN)-based algorithm for precise identification and quantification of bacterial cells using dark field microscopy. Results The results obtained from spike-in and clinical sample analyses demonstrated the accuracy, high sensitivity (with a detection limit of 10 CFU/mu L), and time-saving nature (30 min) of our tM13-based immunomagnetic enrichment approach combined with AI-enabled analytics, thereby supporting its potential to facilitate the identification of diverse E. coli strains in complex samples. Conclusion The study established a rapid and accurate detection strategy for E. coli utilizing trun

关键词： Escherichia coli Truncated M13 bacteriophage Convolutional neural network rapid quantification Dark field microscopy

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Optimization and validation of HPLC-DAD method for simultaneous analysis of sweeteners, preservatives, and caffeine in sugar-free beverages

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EUROPEAN FOOD RESEARCH AND TECHNOLOGY 2023年第11期249卷 2797-2805页

作者： Szekelyhidi, Rita Ajtony, Zsolt Lakatos, Erika Hegyi, Oliver Sik, Beatrix Szecheny Istvan Univ Albert Casimir Fac Mosonmagyarovar Dept Food Sci 15-17 Lucsony St H-9200 Mosonmagyarovar Hungary

In this study, the optimization and performance evaluation of a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for the simultaneous determination of four sweeteners (acesulfame-potassium, saccharin, aspartame, and rebaudioside A), two preservatives (sodium benzoate and potassium sorbate), and caffeine in sugar-free drinks are presented. The separation was carried out using a gradient elution of acetonitrile and phosphate buffer (12.5 mM, pH = 3.3) on a reversed-phase column. Under the optimized conditions, the method allowed for the specific and selective simultaneous separation of all target analytes in less than 9 min. The developed HPLC-DAD method was validated and demonstrated excellent linearity (all analytical curves showed R-2 & GE;0.9995), satisfactory accuracy (recovery values ranging between 94.1 and 99.2% in real samples), and repeatability (intra- and inter-day relative standard deviations were & LE;2.49%). Overall, 69 products available in the Hungarian market were successfully tested with the applied method.

关键词： Sweeteners Preservatives Caffeine HPLC-DAD Drinks rapid quantification

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A biomolecule compatible electrical model of microimpedance affinity biosensor for sensitivity improvement in cell detection

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SENSORS AND ACTUATORS A-PHYSICAL 2010年第2期157卷 280-289页

作者： RoyChaudhuri, C. Das, R. Dev Bengal Engn & Sci Univ Shibpur Dept Elect & Telecommun Engn Howrah 711103 W Bengal India

Microimpedance affinity biosensors using interdigitated electrodes are being extensively employed for label free and rapid detection of bacteria molecules. To improve the sensitivity of such sensors, it is required to maximize the antibody immobilization and bacteria captured for an applied input concentration through proper quantification and also optimize the electrode geometry to achieve maximum signal to noise ratio. In this paper, a biomolecule compatible electrical model has been developed which quantifies the number of immobilized antibody and captured bacteria by simple calculations from impedance spectroscopy results which are comparable to the accuracy obtained by conventional time consuming and sometimes expensive biochemical methods and can also estimate the electrode impedance associated with each biomolecule. The model has been applied for the detection of Escherichia coli K12 bacteria by antibody-antigen binding method on gold electrodes with PBS as the background solution. The quantitative estimation of the actual number of antibody immobilized and bacteria captured for input concentration of 10(3) and 10(6) CFU/ml on the gold surface in this case has been matched with the conventional optical density methods and radioactive labeling method respectively. A quantitative relationship between the geometrical parameters and the output signal has been deduced after extracting the electrode impedance for each biomolecule and the effects of transducer geometry on the sensitivity of this biosensor have been matched with recently reported experimental results for the same system. Thus the proposed model enables quantitative estimation of the antibody immobilized and bacteria captured and quantitative prediction of the optimum geometry for maximum sensitivity eliminating the need for trial and error experimentation. This will lead to a rapid and cost effective method for performance enhancement of microimpedance biosensors with different topologies. (C) 2009 E

关键词： Biomolecule compatible electrical model rapid quantification Impedance biosensor Electrode geometry

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rapid Determination of Icing Inhibitor (Ethyl Glycol) in Aviation Fuel by Near-infrared Spectroscopy

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PETROLEUM SCIENCE AND TECHNOLOGY 2009年第1期27卷 12-19页

作者： Wang, J. Xing, Z. Shen, G. Ye, Y. Naval Aeronaut & Astronaut Univ Dept Aerocraft Engn Yantai 264001 Shandong Peoples R China

A method for determination of ethyl glycol in aviation fuel by near-infrared spectroscopy is presented. The near-infrared signal of ethyl glycol was analyzed and the wavelength range from 960 to 980 nm and a signal processing method were chosen. A calibration model for ethyl glycol was established by partial least squares regression. The calibration model was satisfactory with parameters of R2 = 0.9719, predictive residual error sum of squares equal to 0.007, and standard error of calibration equal to 0.010%. Statistical t-tests ( = 0.05) verified that there was no significant difference between the near-infrared model and the reference method using refraction analysis according to ASTM D5006. Repeatability testing showed that the near-infrared model has precision equivalent to that of the reference method. The study indicates that near-infrared spectroscopy represents a simpler, more rapid, and highly effective test for ethyl glycol in aviation fuel than conventional quantitative analysis.

关键词： aviation fuels ethyl glycol icing inhibitors near-infrared spectroscopy rapid quantification

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Assessment of atmospheric solids analysis probe as a tool for the rapid determination of drug purity

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DRUG TESTING AND ANALYSIS 2024年第8期16卷 807-816页

作者： Mathias, Simone Burns, Daniel Hambidge, Tabatha McCullough, Bryan J. Hopley, Chris J. Douce, David Sage, Ashley Sears, Patrick Univ Surrey Sch Chem & Chem Engn Guildford Surrey England LGC Natl Measurement Lab Teddington England Waters Corp Wilmslow England Univ Surrey Sch Chem & Chem Engn Guildford GU2 7XH Surrey England

The ability to determine the purity (% controlled compound) of drug-of-abuse samples is necessary for public health and law enforcement. Here, we describe the assessment of atmospheric solids analysis probe (ASAP) for the rapid determination of drug purity for a set of formulated pharmaceuticals, chosen due to their availability, uncontrolled status and consistency. Paracetamol and loratadine were used as models of high and low purity compounds being similar to 90% and similar to 10% active ingredient, respectively. Individual tablets were ground up and diluted in an internal standard solution. The resulting samples were analysed by ASAP coupled to a Waters QDa mass spectrometer followed by confirmatory testing by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The inclusion of a non-matched internal standard (quinine) improved linearity and repeatability of drug analysis by ASAP-MS. Levels of drug purity using formulated pharmaceutical tablets were found to be highly comparable with results produced by the 'gold standard' LC-MS/MS technique. rapid determination of drug purity is therefore possible with ASAP-MS for highly concentrated samples with minimal sample preparation. It may be possible to use this deployable system to determine drug purity outside of a laboratory setting.

关键词： ambient ionisation ASAP atmospheric solids analysis probe drug purity rapid quantification

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Quantifying Nanoplastics in Soil-Cultured Plants Based on a Microcombustion Calorimeter

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ENVIRONMENTAL SCIENCE & TECHNOLOGY LETTERS 2023年第11期10卷 1130-1134页

作者： Chen, Hanwen Zhang, Xin Xing, Shuping Hao, Zhipeng Chen, Baodong Zhu, Yong-Guan Chinese Acad Sci Res Ctr Ecoenvironm Sci State Key Lab Urban & Reg Ecol Beijing 100085 Peoples R China Univ Chinese Acad Sci Beijing 100049 Peoples R China Chinese Acad Sci Key Lab Urban Environm & Hlth Inst Urban Environm Xiamen 361021 Peoples R China

Nanoplastics have been detected in a variety of plants and threaten plant growth. To further investigate the physiological damage of nanoplastics to plants and their translocation in plants, it is crucial to quantify the nanoplastics in the plant. However, until now, no studies have reported on how to quantify nanoplastics in soil-cultured plants. Here, we proposed to determine the polyethylene (PE) and poly-(methyl methacrylate) (PMMA) contents in soil-cultured plants using a microcombustion calorimeter (MCC). Since the thermal properties of nanoplastics were different from those of plants and soil, this method was not affected by the environmental background. The linear relationships were developed between the sample total heat release (THR) and nanoplastic proportions. Generally, the application of MCC to quantify PE and PMMA resulted in low detection limits (LODs), quantification limits (LOQs), and high spiked recoveries. The method also achieved a high level of accuracy (relative standard deviation and A type uncertainty), demonstrating the feasibility of the proposed method. This is the first report to quantify nanoplastics in soil-cultured plants based on MCC. It provides the possibility for rapid quantification of nanoplastics in plants and thus allows in-depth research of the behavior of nanoplastics in a soil-plant system.

关键词： Soil-cultured plants nanoplastic content rapid quantification microcombustion calorimeter total heat release

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Novel detection method to rapidly quantify toxic cucurbitacin in Lagenaria siceraria (bottle gourd)

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JOURNAL OF FOOD SCIENCE AND TECHNOLOGY-MYSORE 2023年第1期60卷 160-170页

作者： Saurabh, Chaturbhuj Kumar Ghosh, Sunil Kumar Sanyal, Bhaskar Bhabha Atom Res Ctr Food Technol Div Mumbai 400085 Maharashtra India

Bottle gourd (Lagenaria siceraria) is widely used in India for its medicinal properties. Sometimes its consumption as a fruit or juice can lead to serious health issues due to the presence of toxic cucurbitacins (Cuc) like Cuc B at high concentration. In the present study, a molecularly imprinted polymer (MIP) based method was developed to quantify Cuc B in commercial bottle gourd juice. Cuc B quantification was also done in peel and pulp of fresh bottle gourd fruit. Developed MIP was superior to NIP (non-imprinted polymer) in terms of equilibrium binding, solid-phase extraction, selectivity, and loading capacity of Cuc B. Scatchard plot analysis, adsorption kinetics, and surface area study further strengthen the fact that prepared imprinted polymer was better than NIP for Cuc extraction. TLC and high-resolution LC-MS based analysis revealed that MIP selectively extracts Cuc B from all the studied samples. It was further observed that gamma irradiation and beta-glucosidase treatment did not have any significant (p > 0.05) effect on Cuc concentration. The quantity of Cuc in 100 g of juice, peel, and pulp was 2.81 +/- 0.26, 3.37 +/- 0.31, and 2.39 +/- 0.21 mg, respectively. Analytical method validation indicates that the MIP based method was highly linear, precise, and accurate to rapidly quantify toxic Cuc B in bottle gourd consequently assuring its quality.

关键词： Molecularly imprinted polymer Method development rapid quantification Cucurbitacin Lagenaria siceraria Food quality assurance

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