Development of Magnetic Molecularly Imprinted Polymer Coupled Nanospray Ion Source for Analysis of Cephalosporin Antibiotics in Food Samples.

A magnetic molecularly imprinted polymer (MMIP) coupled nanospray ion source was developed for analysis of cephalosporin antibiotics in food samples. MIP coated Fe3O4 nanospheres were prepared for magnetic solid-phase extraction (MSPE) of the antibiotics in the extract of samples and then integrated into the nanospray capillary for further desorption and mass spectrometry analysis. The developed device combines the advantages of high extraction efficiency of MSPE, unique selectivity of MIPs, and fast analysis speed of ambient ionization mass spectrometry (AIMS). Five cephalosporin antibiotics in milk, egg, and beef samples were analyzed using the developed methods. High sensitivities with limits of detection (LODs) from 0.3 to 0.5 µg kg-1 were achieved for cephalosporin antibiotics in milk, egg, and beef samples, respectively. Good linearity, determination coefficient values (R2 > 0.992), and precision (RSD < 15%) with recoveries ranging from 72.6% to 115.5% were obtained using the spiked milk, egg, and beef sample matrices.

Real-time traceability of sorghum origin by soldering iron-based rapid evaporative ionization mass spectrometry and chemometrics.

Sorghum is an important grain with a high economic value for liquor production. Tracing the geographical origin of sorghum is vital to guarantee the liquor flavor. Soldering iron-based rapid evaporative ionization mass spectrometry (REIMS) combined with chemometrics was developed for the real-time discrimination of the sorghum's geographical origin. The working conditions of soldering iron-based ionization were optimized, and then the obtained MS profiling data were processed using chemometrics analysis methods, including principal component analysis-linear discriminant analysis and orthogonal projection to latent structures discriminant analysis (OPLS-DA). A recognition model was established, and discriminations of sorghum samples from 10 provinces in China were achieved with a correct rate higher than 90%. On the basis of OPLS-DA, the specific ions of m/z 279.2327, 281.2479, and 283.2639 had relatively strong discrimination power for the geographical origins of sorghum. The developed method was successfully applied in the discrimination of sorghum origins. The results indicated that the soldering iron-based REIMS technique combined with chemometrics is a useful tool for direct, fast, and real-time ionization of poor conductivity samples and acquisition of metabolic profiling data.

Tissue distribution study of perfluorooctanoic acid in exposed zebrafish using MALDI mass spectrometry imaging.

Perfluorooctanoic acid (PFOA) as an emerging environmental contaminant, has become ubiquitous in the environment. It is of significance to study bioconcentration and tissue distribution of aquatic organisms for predicting the persistence of PFOA and its adverse effects on the environment and human body. However, the distribution of PFOA in different tissues is a complex physiological process affected by many factors. It is difficult to be accurately described by a simple kinetic model. In present study, a new strategy was introduced to research the PFOA distribution in tissues and estimate the exposure stages. Zebrafish were continuously exposed to 25 mg/L PFOA for 30 days to simulate environmental process. Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) method was used to monitor the spatio-temporal distribution of PFOA in zebrafish tissues. By analyzing the law of change obtained from the high spatial resolution MSI data, two different enrichment trends in ten tissues were summarized by performing curve fitting. Analyzing the ratio of two types of curves, a new "exposure curve" was defined to evaluate the exposure stages. With this model, three levels (mild, moderate, and deep pollution stage) of PFOA pollution in zebrafish can be simply evaluated.

Sulfonamide-Selective Ambient Mass Spectrometry Ion Source Obtained by Modification of an Iron Sheet with a Hydrophilic Molecularly Imprinted Polymer.

We have described a sulfonamide-selective ambient ion source coupled with electrospray ionization mass spectrometry (ESI-MS) for selective extraction and determination of trace sulfonamide antibiotics. It is obtained by modifying an iron sheet with a sulfadiazine-templated hydrophilic molecularly imprinted polymer (SF-HMIP). It behaves as both an online extractor and a MS ion source. Five sulfonamide antibiotics, including sulfamethoxazole (SMZ), sulfamerazine (SMR), sulfisoxazole (SIZ), sulfathiazole (ST), and sulfameter (SMD), were chosen to evaluate SF-HMIP coupled with ESI-MS, which showed good linearity in the range of 0.2-1000 ng/mL with correlation coefficient values (R2) over 0.9946. The limits of detection (LODs) for analysis of pure water and honey were in the range of 0.1-0.2 and 0.2-1.5 ng/mL, respectively. Limits of quantitation (LOQs) for analysis of pure water and honey were in the range of 0.3-0.5 and 1.0-5.0 ng/mL, respectively. The results demonstrated that SF-HMIP combined with ESI-MS could be applied for the direct analysis of five trace sulfonamide compounds in honey and pure water with recoveries ranging from 76 to 129%.

Modeling the distribution of malachite green in zebrafish using matrix-assisted laser desorption/ionization mass spectrometry imaging.

Understanding the spatial distribution of bioactive small molecules is indispensable for elucidating their biological or pharmaceutical roles. Here, a rapid and effective analysis strategy was introduced to study the distribution of veterinary drugs in aquatic products. Malachite green (MG), one of the most widely used veterinary drugs in aquaculture, was selected as the targeted compound. Zebrafish (Danio rerio) was used as a model organism. After an exposure test, the matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) technique was applied to directly analyze the content changes of malachite green in zebrafish tissues. The reliable relationship of exposure time and content change of MG was described precisely by the extended Freundlich equation. The process of modeling was discussed in detail, and some important parameters or trend information was obtained, including the maximum content of MG in different fish tissues, time to maximum content, elimination time, equilibrium content, and so on. With a simplification of sample pretreatment, this research strategy can be used for monitoring the spatial distribution of veterinary drugs and related metabolites of laboratory-exposed fish. The obtained model can provide a perspective for rational drug use in aquaculture and precise drug residue detection in production activities.

Fast construction of core-shell structured magnetic covalent organic framework as sorbent for solid-phase extraction of zearalenone and its derivatives prior to their determination by UHPLC-MS/MS.

Magnetic covalent organic framework nanocomposite denoted as Fe3O4@TAPB-Tp with core-shell structure was fabricated via a simple template-mediated precipitation polymerization method at mild conditions. The polyimine network shell was created through the polymerization of 1,3,5-tris(4-aminophenyl)-benzene (TAPB) and 1,3,5-triformyl-phloroglucinol (Tp) in tetrahydrofuran (THF) by the Schiff-base reaction. Featuring with large specific surface area (163.19 m2 g-1), good solution dispersibility, and high stability, the obtained Fe3O4@TAPB-Tp exhibited high adsorption capacities and fast adsorption for zearalenone and its derivatives (ZEAs). The adsorption isotherms showed multilayer adsorption dominated at low concentration and monolayer adsorption at high concentration between the interface of ZEAs and Fe3O4@TAPB-Tp. With the Fe3O4@TAPB-Tp as sorbent, a magnetic solid-phase extraction-ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for simultaneous adsorption and detection of five ZEAs in complex samples. The proposed method displayed favorable linearity, low limits of detection (0.003 ~ 0.018 µg kg-1), and good repeatability (2.37~10.4%). The developed method has been applied for real sample analysis, with recoveries of 81.27~90.26%. These results showed that Fe3O4@TAPB-Tp has a good application potential for the adsorption of ZEAs in food samples. Magnetic covalent organic framework nanocomposite (Fe3O4@TAPB-Tp) were quickly fabricated at mild conditions and used as effective adsorbent for magnetic solid-phase extraction of zearalenone and its derivatives (ZEAs) from food samples prior to ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis.

[Research progress of solid phase extraction materials in the application of metal ion pretreatment].

Monitoring of trace heavy metal pollutants released during industrial and agricultural processes is essential because of their widespread distribution in the environment and health hazards. Several techniques, including inductively coupled plasma-mass spectrometry (ICP-MS), inductively coupled plasma-optical emission spectrometry (ICP-OES), electrothermal atomic absorption (ETAAS), and flame atomic absorption spectrometry (FAAS), have been proposed for the determination of heavy metals in serum, plasma, whole blood, and food. All these techniques have earned robust recognition in the field of trace heavy metals and have many advantages such as multi-elemental analysis capability, large dynamic linear range, low detection limits, and high productivity. Nevertheless, most of the recommended techniques require digestion of the sample and extraction with an organic solvent for isolation of the metal ion from the sample solution prior to analysis. Despite improvements in the performance of modern analytical instruments, the direct determination of heavy metal ions in real samples is difficult because of their low concentration levels and matrix interference. Thus, extraction and clean-up steps are required for pre-concentration of the analyte, so that detection and elimination of the interfering matrix component are possible. Solid-phase extraction (SPE) is one of the popular metal ion pretreatment methods. The advantages of SPE include easy cartridge/column regeneration, high analytical frequency, and high preconcentration factors for sorbents with high adsorption capacities. On the other hand, when the analytes are extracted from a complex matrix such as serum and meat samples, large amounts of proteins from the samples can be retained on the sorbent surface, obstructing the binding sites on the sorbent and leading to poor precision and accuracy. The key to metal ion detection is the development of new SPE materials with high efficiency and enrichment factors as well as an effective pretreatment technology. Nanomaterials such as restricted-access carbon nanotubes, nanoadsorbents, nanoparticle carriers, and magnetic nanoparticles have shown great promise in advancing biomedical and environmental analysis because of the unique properties originating from their ultrafine dimensions. Nanomaterials can provide large specific surface areas and tunable functional groups to facilitate metal ion absorption. They could also possess superior optical properties and allow for high sensitivity in simple fluorescent or colorimetric detection methods. Owing to their excellent mechanical and chemical stability, polymer materials have been of great interest as adsorbents for the SPE of metal ions from solution. Moreover, a designed polymeric material can show triple functionality such as physical adsorption, chelate formation, and ion exchange for the target metal ions. A dual-functional nanomaterial-DNAzyme platform can simultaneously allow for the sensitive detection and effective removal of heavy metal ions in water. Thus, this platform can serve as a simple, cost-effective tool for rapid and accurate metal quantification in the determination of human metal exposure and inspection of environmental contamination. Furthermore, the new photocaged chelator can uncage and release the combined metal ions into an aqueous solution that is free of the other components of the matrix. In this manner, we can develop diagnostic tests for metal ions that are often difficult to detect using other methods. In this paper, the characteristics of new SPE materials, including nanomaterials, polymer materials, and functional materials as well as advances in their applications to the preparation of complex samples are summarized, and the direction for future development is proposed.

Differentiation between Fresh and Frozen-Thawed Meat using Rapid Evaporative Ionization Mass Spectrometry: The Case of Beef Muscle.

An intelligent surgical knife (iKnife) coupled with rapid evaporative ionization mass spectrometry (REIMS) was employed for the lipidomic profiling of fresh and frozen-thawed beef muscle. The data were obtained by REIMS and then processed using multivariate statistical analysis methods including principal component analysis-linear discriminant analysis (PCA-LDA) and orthogonal partial least-squares discriminant analysis (OPLS-DA). The discrimination of fresh and frozen-thawed meat has been achieved, and the real-time identification accuracy was 92-100%. Changes in the composition and content of fatty acids and phospholipids were statistically analyzed by OPLS-DA, and the ions of m/z 279.2317, m/z 681.4830, and m/z 697.4882 were selected as differential compounds/metabolites. The developed method was also successfully applied in the discrimination of fresh and frozen-thawed meat samples. These results showed that REIMS as a high-throughput, rapid, and real-time mass spectrometry detection technology can be used for the identification of fresh and frozen-thawed meat samples.

An integrated method for monitoring thermal processing temperature of pork based on Q-Exactive mass spectrometry and chemometrics.

Meat heating endpoint temperature (EPT) is an important indicator to ensure the safety of cooked meat. Accurately determining the EPT of cooked meat and ready-to-eat meat products is an important strategy to ensure food safety. In this study, a comprehensive metabolic method based on UPLC-Q Exactive and chemometrics was developed to study the metabolites differences among pork roasted at different temperatures in order to select markers indicating EPT and discover new toxic heat-induced compounds. A two-step extraction method was applied to avoid the loss of metabolite information caused by sample preparation. Using chemometrics, the five compounds of creatine, creatinine, 2-amino-1-methyl-6-phenylimidazo (4,5-b) pyridine (PhIP), 2-methyl-6-amino-5-hydroxymethylpyrimidine (TMP) and compound with the m/z of 114.04316 were selected as markers, and four of them were further confirmed by chemical standards. It is worth noting that TMP was discovered in roasted pork for the first time. In addition, targeting studies aimed at quantifying the selected markers were conducted at different thermal processing temperatures. From the quantification results, it can be concluded that the heat temperature not exceed 180 °C is recommended to reduce the content of toxic compounds. This study has proved that the integration of UPLC-Q Exactive and chemometrics could provide an efficient method for the study of markers related to thermal process and new toxic heat-induced compounds.

The distribution and changes of glycoalkaloids in potato tubers under different storage time based on MALDI-TOF mass spectrometry imaging.

Glycoalkaloids (GAs) are toxic secondary metabolites in potatoes, which are harmful to human body. The storage time has a great influence on the biosynthesis and distribution of GAs. In present study, an imaging mass microscope (iMScope) was used to investigate the distribution and changes of GAs in potato tubers under different storage time (0, 10, 15, 20, 30, 40 and 60 days). We established a growth model with logistic equation to evaluate the growth trends of four major GAs in sprout, periderm and medulla. The results showed that the growth rate and relative contents of four GAs in sprout and periderm were significantly higher than that in medulla. In addition, four GAs also presented different change trends. For dehydrosolanine and α-solanine, rapid growth period of these two GAs in sprout (about at the day 23, similar to these in medulla) was later than which period in periderm (about at the day 17), while rapid growth of dehydrochaconine and α-chaconine appeared at almost the same time (about at the day 20). Based on the biosynthesis and metabolism of GAs, we have made possible explanations for these results. This study is useful for comprehending the metabolism of GAs in different parts and monitoring food safety in potatoes.

Rapid and sensitive determination of trace fluoroquinolone antibiotics in milk by molecularly imprinted polymer-coated stainless steel sheet electrospray ionization mass spectrometry.

Rapid analysis of trace analytes in complex biological samples is a great challenge for direct mass spectrometry, which suffers from low detection sensitivity. In this study, molecular imprinting technology was explored on the stainless steel sheet and integrated with the electrospray ionization method for direct sample analyses. The molecularly imprinted polymer-coated stainless steel sheet (MIPCS) was prepared and used as a solid-phase microextraction tip for rapid sampling of trace fluoroquinolone antibiotics in milk samples and then applied as an electrospray ionization tip to couple MS for sensitive detection. Our results shown that MIPCS could significantly enrich the trace fluoroquinolone antibiotics in milk samples. In our study, the extraction process of milk sample was completed within 30 min and the direct MS analysis was accomplished within 1 min. In addition, this proposed MIPCS-ESI-MS method showed a good linearity (R2>0.99) ranged from 1 to 1000 ng mL-1. The limits of detection (LODs) and limits of quantitation (LOQs) for the analytes range from 0.1 to 5 ng mL-1. The recoveries were in a range of 78.84%-103.04%. The relative standard deviation (RSD%) of inter-day and intra-day precision ranged from 7.00% to 10.4% and 4.46%-11.44% respectively. Overall, the proposed MIPCS-ESI-MS method could be feasibly used as a rapid and sensitive method for determination of trace analytes in complex food samples.

Magnetic solid phase extraction sorbents using methyl-parathion and quinalphos dual-template imprinted polymers coupled with GC-MS for class-selective extraction of twelve organophosphorus pesticides.

A novel magnetic dual-template molecularly imprinted polymer (DMIP) was prepared with methyl-parathion and quinalphos as templates. For comparison, a series of single-template polymers with only methyl-parathion (MPMIP) or quinalphos (QPMIP) as template as well as a non-imprinted polymer (NIP) in the absence of the template, were synthesized using the same procedure of DMIP. The obtained MIPs were characterized by scanning electron microscopy(SEM), Fourier transform infrared (FT-IR) spectroscopy, vibrating sample magnetometer (VSM), and X-ray diffraction (XRD). The properties including kinetic effect, thermodynamic effect, selectivity, and reusability of MIPs were investigated . Only DMIP possessed high affinity and good recognition for all twelve OPPs including quinalphos, isazophos, chlorpyrifos-methyl, chlorpyrifos, methidathion, triazophos, profenofos, fenthion, fenitrothion, methyl-parathion, parathion, and paraoxon in comparison to MPMIP, QPMIP, or NIP. Moreover, DMIP was used as magnetic solid phase extraction (MSPE) sorbent for the pre-concentration of twelve OPPs in cabbage samples. The developed DMIP-MSPE-GC-MS method showed high sensitivity, low LODs (1.62-13.9 ng/g), fast adsorption equilibrium (10 min), and acceptable spiked recoveries (81.5-113.4%) with relative standard deviations (RSD) in the range 0.05-7.0% (n = 3). The calibration plots were linear in the range 10-800 ng/mL with coefficients of determination (R2) better 0.99 for all twelve compounds. These results suggest that the DMIP is applicable for rapid determination and high throughput analysis of multi-pesticide residues. Graphical abstract.

Liquid-Phase Ion Trap for Ion Trapping, Transfer, and Sequential Ejection in Solutions.

In this study, a new method/mechanism to manipulate ions in solution was developed, based on which liquid-phase ion trap was built. In this liquid-phase ion trap, ion manipulations conventionally performed in a quadrupole ion trap or in a trapped ion mobility spectrometer placed in a vacuum were achieved in solutions. Through theoretical derivation and numerical simulation, it is found that ions have different motional characteristics than those in vacuum. Instead of a radio frequency quadrupole electric field, tunable DC electric fields together with a constant liquid flow were applied to control ion motions in solution. Different ions could be trapped and focused in a potential well, and ion densities could be increased by over 100-fold. By adjusting the DC electric field of the potential well, trapped ions could be transferred into another trapping region or sequentially released for detection. Ions released from the liquid-phase ion trap were then detected by a mass spectrometer interfaced with an electrospray ionization source. Since the ion manipulation mechanism in solution is different and complementary to that in vacuum, the use of a liquid-phase ion trap could also boost detection sensitivity and the mixture analysis capability of a mass spectrometer.

The Coupling of Taylor Dispersion Analysis and Mass Spectrometry to Differentiate Protein Conformations.

Measuring the conformations of protein and protein-ligand complexes in solution is critical for investigating protein bioactivities, but their rapid analyses remain as challenging problems. Here, we report the coupling of Taylor dispersion analysis (TDA) with mass spectrometry (MS) for the rapid conformation differentiation of protein and noncovalent protein complex in solution environments. First, a branched capillary design was applied to achieve double band detection for the peak retention time correction in TDA measurements. After ionization, analytes were further detected and distinguished by their mass to charge (m/z) ratios in the consequent MS analysis. As a result, protein or protein complex in a mixture could be analyzed in terms of both hydrodynamic radius and m/z. The feasibility of this method was verified by analyzing a mixture of angiotensin II and phenylalanine, and the conformations of cytochrome C at different pH conditions were then investigated. As proof-of-concept demonstrations, the complexes of tri-N-acetylchitotriose with two proteins (lysozyme and cytochrome C) were characterized with results verified by molecular dynamics simulations. The TDA-MS method is promising for rapid structural analyses of trace amounts protein-ligand complexes, which could potentially be used to differentiate intact protein or protein complex conformations.

[Research advances in imaging technology for food safety and quality control].

Food quality and safety are issues of concern to the government, food industry, and consumers; hence, it is imperative to detect harmful substances in foodstuff. Traditional techniques for this purpose include biochemical methods and instrumental analysis methods such as chromatography and chromatography-mass spectrometry. These methods, however, are time-consuming and unable to obtain the spatial distribution of the analytes. Therefore, the development of rapid, non-destructive, real-time, and visual detection technologies has emerged as a hotspot in the field of food research. In recent years, hyperspectral imaging, which combines imaging and spectral technology, is rapidly gaining ground. This technique allows one to determine the geometrical characteristics and chemical composition of samples. Compared with traditional spectral technologies, hyperspectral imaging has the advantages of wide detection ranges, in addition to being real-time and non-destructive. At present, hyperspectral imaging is widely used in meat quality evaluation, detection of adulteration, and meat classification. In addition, Raman imaging is mainly used for the detection of illegal additives in food and for adulteration detection. This technology is fast, non-destructive, and low cost; furthermore, spectral and spatial information of the targets can be simultaneously obtained. Mass spectrometry imaging allows for the visualization and high-throughput analysis of sample tissues, without the need for complex sample preparation steps such as labeling and staining. Compared with other imaging technologies, mass spectrum information of substances can be obtained by mass spectrometry imaging. As a molecular visualized technology, it helps obtain the spatial distribution of nutrients and harmful substances in food. Mass spectrometry imaging has unique advantages in food research, e. g., it is used for molecular-level detection and accurate positioning of substances, and hence, it has excellent application prospects in this field. In this paper, recent literature data about imaging technologies in the field of food research, including 72 reports published in professional local and overseas magazines, are collated. The principles of hyperspectral imaging, Raman imaging, and mass spectrometry imaging are introduced, along with the detailed applications of these methods in the quality detection, source identification, and microbial pollution of food. In addition, it also includes food physical damage, food adulteration and food chemical residues. Besides, the advantages and disadvantages of these imaging technologies are discussed. Finally, prospects for the development of imaging technologies in food research are presented. Future work related to hyperspectral imaging should focus on the development of high-sensitivity cameras and high-resolution systems. Improving the data processing efficiency and adding prediction models are also key points for the future. Future studies on Raman imaging can focus on the application of different chemometrics algorithms that would improve the evaluation of food quality and safety parameters. Expanding the scope of application of these methods in food research will also be the focus of future research. Regarding mass spectrometry imaging, attempts should be made to improve the ionization methods, detection sensitivity, spatial resolution, and data processing effectiveness. Additionally, the combination of spectral imaging and mass spectrometry imaging gives full play to their advantages, so that spectral and mass spectrometry information of the targets can be obtained. In short, the application of imaging technologies in food research is expected to be more promising.

Structural Analysis of Biomolecules through a Combination of Mobility Capillary Electrophoresis and Mass Spectrometry.

The 3D structures of biomolecules determine their biological function. Established methods in biomolecule structure determination typically require purification, crystallization, or modification of target molecules, which limits their applications for analyzing trace amounts of biomolecules in complex matrices. Here, we developed instruments and methods of mobility capillary electrophoresis (MCE) and its coupling with MS for the 3D structural analysis of biomolecules in the liquid phase. Biomolecules in complex matrices could be separated by MCE and sequentially detected by MS. The effective radius and the aspect ratio of each separated biomolecule were simultaneously determined through the separation by MCE, which were then used as restraints in determining biomolecule conformations through modeling. Feasibility of this method was verified by analyzing a mixture of somatostatin and bradykinin, two peptides with known liquid-phase structures. Proteins could also be structurally analyzed using this method, which was demonstrated for lysozyme. The combination of MCE and MS for complex sample analysis was also demonstrated. MCE and MCE-MS would allow us to analyze trace amounts of biomolecules in complex matrices, which has the potential to be an alternative and powerful biomolecule structure analysis technique.

Mobility Capillary Electrophoresis-Restrained Modeling Method for Protein Structure Analysis in Mixtures.

Protein stereostructure analysis in mixtures still remains challenging, especially large-scale analysis such as in proteomics. With the capability of measuring the hydrodynamic radius of ions in the liquid phase, mobility capillary electrophoresis (MCE) has been applied to study the structure of peptides. In this study, MCE was extended for protein mixture separation and their corresponding hydrodynamic radius analyses. After ellipsoid approximation, the results obtained by MCE experiments were then used as a restraint in molecular dynamics simulations to predict the most probable structure of each protein. Besides a three-protein mixture, a mixture of disulfide bond reduced insulin was also studied by this MCE-restrained modeling method. The results obtained by this method agree with literature studies, and mass spectrometry experiments were also carried out to confirm our findings.

Rapid characterization of structure-dependency gas-phase ion/ion reaction via accumulative tandem MS.

To enable the rapid detection of biomolecule reactivity and reaction sites, we developed a method based on gas-phase ion/ion reaction and accumulative tandem mass spectrometry (MS). Structure-dependency reactions in gas-phase were performed between biomolecule ions and their reaction partner ions with opposite polarities in a quadrupole ion trap. Gas-phase peptide bioconjugation with pyridoxal-5-phosphate (PLP) was chosen as a proof-of-principle example. It is found that the Coulomb attraction force holds reaction partners close together, which increasing the reaction probability. Post reaction, reaction sites were identified by the consequent accumulative tandem MS method, in which informative product ions in low abundance were enriched by more than 100 times in another quadrupole ion trap. With enough product ions, tandem MS was performed, and reaction sites could be identified unambiguously. Since those reactions are normally biomolecular structure dependent, density functional theory (DFT) calculations were also carried out to understand the reaction mechanism. The method allows for rapid characterization of structure dependent reactivity of a biomolecule, and opens a new avenue for drug development and biomolecule structure analyses.

A two-step method for rapid characterization of electroosmotic flows in capillary electrophoresis.

The measurement of electroosmotic flow (EOF) is important in a capillary electrophoresis (CE) experiment in terms of performance optimization and stability improvement. Although several methods exist, there are demanding needs to accurately characterize ultra-low electroosmotic flow rates (EOF rates), such as in coated capillaries used in protein separations. In this work, a new method, called the two-step method, was developed to accurately and rapidly measure EOF rates in a capillary, especially for measuring the ultra-low EOF rates in coated capillaries. In this two-step method, the EOF rates were calculated by measuring the migration time difference of a neutral marker in two consecutive experiments, in which a pressure driven was introduced to accelerate the migration and the DC voltage was reversed to switch the EOF direction. Uncoated capillaries were first characterized by both this two-step method and a conventional method to confirm the validity of this new method. Then this new method was applied in the study of coated capillaries. Results show that this new method is not only fast in speed, but also better in accuracy.

Dual-Polarity Ion Trap Mass Spectrometry: Dynamic Monitoring and Controlling Gas-phase Ion-Ion Reactions.

A dual-polarity linear ion trap (LIT) mass spectrometer was developed in this study, and the method for simultaneously controlling and detecting cations and anions was proposed and realized in the LIT. With the application of an additional dipolar DC field on the ejection electrodes of an LIT, dual-polarity mass spectra could be obtained, which include both the mass-to-charge (m/z) ratio and charge polarity information of an ion. Compared with conventional method, the ion ejection and detection efficiency could also be improved by about one-fold. Furthermore, ion-ion reactions within the LIT could be dynamically controlled and monitored by manipulating the distributions of ions with opposite charge polarities. This method was then used to control and study the reaction kinetics of ion-ion reactions, including electron transfer dissociation (ETD) and charge inversion reactions. A dual-polarity collision-induced dissociation (CID) experiment was proposed and performed to enhance the sequence coverage of a peptide ion. Ion trajectory simulations were also carried out for concept validation and system optimization. Graphical Abstract ᅟ.