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Monday
Urinary Glycoprotein Biomarker Discovery for Human Bladder Cancer Using Multi-lectin Affinity Chromatography and LC-MS/MS
| Session: Proteomics: Biomarker Discovery I |
Code: MPC |
Poster: 082 |
Na Yang1; Shun Feng1; Huy Vuong1; Steve Goodison2; Charles J. Rosser2; Fan Xiang3; David M. Lubman1
1University of Michigan, Ann Arbor, MI; 2University of Florida, Jacksonville, FL; 3Shimadzu Biotech, Pleasanton, CA |
Bladder cancer is the fifth-most common malignancy in the world. Early clinical diagnosis of bladder cancer remains a major challenge and the development of noninvasive urinalysis is desirable for both patients and health care providers. The current method for the noninvasive detection of bladder cancer is voided urine cytology (VUC) with poor sensitivity (25-40%). Urinary proteomics is one of the rapidly-growing sub-disciplines of clinical proteomics. Glycoproteins have been recognized as one rich source for diagnostic biomarkers due to the critical roles they play in biological functions and disease progression. To discover potential biomarkers for bladder cancer, novel methodologies for urinary proteome profiling by our group were further optimized and applied to the large-scale identification of human urinary glycoproteins.
Identification of In Vitro Modified Lipoproteins Using MALDI Tandem Mass Spectrometry and a Reduced Proteins Database Approach
| Session: Proteomics: Biomarker Discovery I |
Code: MPC |
Poster: 092 |
Omar Belgacem1; Helen Montgomery2; Matthew Openshaw1; Wu Zidian1; Sobal Grazyna3; Gerald Stubiger4
1Shimadzu Biotech, Manchester, UK; 2Shimadzu, Koichi Tanaka MS Research Laboratory, Manchester, UK; 3Department of Nuclear Medicine, Medical University, Vienna, Austria; 4Department of Vascular Biology Thrombosis Research, Vienna, Austria |
Most of the post translational modifications of peptides are present in the major search engine algorithms like mascot. However, the detection of specific though important modifications like non-enzymatic glycation and N-linked glycosylation is not possible using standard search engines. It requires the use of specific software with their own MSMS library and is very time consuming with large set of MSMS data. The presented work is an efficient method allowing searching for modified peptides within a large set of MSMS Data. The modifications are either defined by the user or in a populated database. The idea is to identify the non-matched matched peptides and direct them towards a designed module that will scan each MSMS spectra under certain constraints.
A New Approach for Accurate Mass Top-Down Sequencing of Intact Proteins/Toxins Using High Resolution Ion-Trap TOF Mass Spectrometry
| Session: Instrumentation: New Concepts I |
Code: MPF |
Poster: 138 |
Christopher Nixon1; Jesse Hines2; Timothy R. Croley1
11Commonwealth of Virginia, DCLS, Richmond, VA; 2Shimadzu Scientific Instruments, Research Triangle Park, NC
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Intact protein measurement by ESI generally requires deconvolution of the multiple-charged ion series. This requires discrimination of signal from background and the presence of a distinguishable charge series. These requirements are not met when there are only a few peaks observed from the charge series and also in mixtures where charge series overlap. Fragmentation of highly-charged precursors also results in highly-charged products that may not have a charge series. In these cases, resolving the isotopes is critical for accurate molecular weight calculation. Classically, every attempt is made to increase signal to obtain a clear charge series. This new approach actually attempts to minimize the number of ions in the trap to minimize the energy spread and increase resolution.
Identification of Pigments Precursors in Garlic Greening by LCMS-IT-TOF System
| Session: Small Molecule Analysis I |
Code: MPG |
Poster: 155 |
Jing Dong1; Dan Hu2; Leren Wan1; Yuki Hashi1; Guanghua Zhao2
1Shimadzu International Trading (Shanghai) Co., Ltd., Beijing , China ; 2China Agricultural University, Beijing, China |
2-(1H-pyrrol-1-yl)succinic acid (P-Asp) and 2-(1H-pyrrol-1-yl)pentanedioic acid (P-Glu) possess similar structures to that of 2-(3,4-dimethylpyrrolyl)-3-methylbutanoic acid (PP-Val), a possible pigment precursor for garlic greening. Therefore, P-Asp and P-Glu were synthesized as model compounds to study their effects on garlic greening. Puree of freshly havested garlic bulbs turned green after being soaked in solutions of P-Asp and P-Glu. By further study on this phenomena, it was found that yellow pigments (which are possible precursors of garlic green) can be produced by mixing the two model compounds respectively with pyruvic acid (component in garlic) at room temperature. Two new yellow pigments from these two reacting systems were purified and characterized using LCMS-IT-TOF, a hybrid mass spectrometer combining ion trap with time-of flight.
High Performance Liquid Chromatography Combined with Multi-stage Mass Spectrometry Analysis of DNA Adducts of Aristolochic Acids
| Session: Small Molecule Analysis I |
Code: MPG |
Poster: 166 |
Haiyan Gao1; Jing Dong3; Feng Feng2; Leren Wan3; Yuki Hashi3; Hailin Wang2
1Central South University, Changsha, China; 2Research Center of Eco-Environmental Sciences, Beijing, China; 3Shimadzu International Trading (Shanghai) Co., Ltd, Beijing, China
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8-methoxy-6-nitro-phenanthro-(3,4-d)-1,3-dioxolo-5- carboxylic acid (AAI) and 6-nitro-phenanthro- (3,4-d)-1,3-dioxolo-5-carboxylic acid (AAII) are the two major components of aristolochic acid. It has been confirmed that aristolochic acid (AA) can react with DNA in human body to form covalent adducts, which can be used as biomarkers of organism exposure to AA and for investigation of the mutagenic and carcinogenic potential of AA. Therefore, it is necessary to develop a sensitive analytical method which can confirm the presence of AA-DNA adducts in vivo and in vitro biological systems. In recent times, the ability of mass spectrometry (MS) to provide structural information has permitted LC/MS to dramatically gain in popularity, becoming essential in the characterization of DNA adducts.
Boundary Element Method for Printed Circuit Board Ion Trap Structure Optimization with the Elimination of Certain Multipole Harmonics
| Session: Instrumentation: Quadupoles and Traps |
Code: MPH |
Poster: 197 |
Chuan-fan Ding1; Gong-yu Jiang1; Li Ding2
1Fudan University, Shanghai, China; 2Shimadzu Research Lab, Shanghai, China |
Compared with cylindric and hyperbolic ion trap, planar shaped mass analyzers such as Rectilinear Ion Trap, Printed Circuit Board Iion Trap(PCBIT) or Halo Trap, always have more complex distribution of multipole harmonics, which affect the efficiency of resonant excitation and ejection of an ion trap at certain beta or q value. Higher level multipole harmonics such as A10 or A14 have complex nonlinear resonant line structure in the stability diagram, easily to create ghost peaks or severe chemical shift. By using boundary element method, series structure of PCBIT with certain twelvepole components with no A10 or A14 components arbitrary structure was found, which were expected with less ghost peaks, ejecting struggling around the outlet slit/hole and better resolution performances.
PCB Ion Trap Mass Spectrometer Coupled with Electrospray Ionization Source
| Session: Instrumentation: Quadupoles and Traps |
Code: MPH |
Poster: 198 |
Gong-yu Jiang1; Xiao-xu Li1; Chan Luo1; Chuan-fan Ding1; Li Ding2
1Fudan University, Shanghai, China ; 2Shimadzu Research Lab, Shanghai, China
|
A mass spectrometer with an electrospray ionization source was comprised of a linear ion trap mass analyzer fabricated by print circuit boards (PCBIT) coupled with atmospheric pressure interface. By altering the potential distribution on the PCB plate pattern for digital RF waveform applied on the ESI-PCBIT, Unit resolution was gained at a scan rate of 2500Th/s while reduced scan rates gave better resolution. The collision-induced dissociation (CID) capabilities of the RIT instrument were evaluated by medicine precursor, peptides, polymers and non-covalent complexes. Overall CID efficiencies of more than 50% were reached, while isolation of an ion with unit resolution at m/z 426 was achieved with high rejection (>98%) of the adjacent ions.
High Precursor Ion Isolation in a Pure Quadrupole Field
| Session: Instrumentation: Quadupoles and Traps |
Code: MPH |
Poster: 199 |
Roger Giles; Matthew C. Gill
Shimadzu Research Laboratories (Europe) Ltd., Manchester, UK |
Ion trap mass spectrometers, of 3D or linear type, commonly have apertures within the trapping electrodes for ion ejection or introduction. It is known that the presence of these apertures cause ‘field faults’ which strongly influence the resonant ejection process. Good analytical performance can be achieved by either geometry modification or by the location of an external electrode in close proximity of the apertures to compensate for the field fault. However, if a linear ion trap is to be employed solely for the purposes of CID and precursor ion isolation no apertures are necessary. Accordingly, it is useful to investigate the process of resonant ejection in a pure quadrupole field.
None-resonant Collision Induced Dissociation in a Digital Linear Ion Trap Time-of-Flight Mass Spectrometer
| Session: Instrumentation: Quadupoles and Traps |
Code: MPH |
Poster: 200 |
Matthew C. Gill; Roger Giles
Shimadzu Research Laboratories (Europe) Ltd., Manchester, UK
|
Collision induced dissociation (CID) without resonant excitation has not been previously demonstrated using an ion trap mass spectrometer. A digital linear ion trap mass spectrometer is described which incorporates the facility to perform collision induced dissociation by accelerating ions along the axis using energies of up to 1 keV. Ions may be trapped, isolated and dissociated, and the daughter ions passed to a time-of-flight mass analyser. Ion optical simulations of the process are compared to experiment. Fragmentation of a range of compounds has been demonstrated.
Design and Fabrication of Ceramic Linear Ion Trap with Its Detector
| Session: Instrumentation: Quadupoles and Traps |
Code: MPH |
Poster: 201 |
Hui Mu; Tao Lin; Xiaohui Yang; Junsheng Zhang; Li Ding
Shimadzu Research Laboratory, Shanghai , China |
A linear ion trap (LIT) can be made in various forms, eg. with four hyperbolic rods, four planar metal plates (RIT) or the one made of printed circuit boards (PCBs). While enjoying the common advantage of larger charge capacity, a PCBs linear ion trap may lead to the low cost production and allows field adjustment after production. Considering ceramic plates have much higher mechanical strength than epoxy resin PCBs, a ceramic PCB ion trap was designed and its fabrication process was investigated. The whole ion optical system, including LIT itself and ion detector has been optimized using SIMION 8.0 and our own programs
Highly Effective Injection Method of MALDI Ions into the Digital Ion Trap
| Session: Instrumentation: Quadupoles and Traps |
Code: MPH |
Poster: 202 |
Kei Kodera; Makoto Hazama; Sadanori Sekiya; Shinichi Iwamoto; Koichi Tanaka
Shimadzu Corporation, Kyoto, Japan |
Digital Ion Trap (DIT) is driven by high voltage with rectangular waveform. For mass scan, the frequency of the waveform is scanned with fixed amplitude. Previously, we developed a MALDI-DIT-MS and evaluated the performance. In addition, we devised a new method of additional ion injection while keeping previously trapped ions in the ion trap. This method was available for MALDI ion accumulation and calibrant injection in the ion trap. However this method was applied to additional ion injection only and not initial injection. In the present study, we modified the method for initial ion injection from MALDI ion source into the DIT. We evaluated the method compared to conventional rapid RF startup method using computer simulations and experiments.
Accelerating Lipid Profiling of Human Samples for Biomarker Discovery Using UFLC-IT-TOF
| Session: Lipids: Methods/Profiling |
Code: MPJ |
Poster: 252 |
Simon Ashton1; Neil J. Loftus1; Chris Titman1; Albert Koulman2
1Shimadzu, Manchester, UK; 2MRC Human Nutrition Research, Cambridge, UK |
Our goal is the development of faster and more robust lipidomics methods compatible with sample sets (>500) without compromising the detection of reliable ion signals. High sample throughput demands limited sample clean-up and short analysis times. The robustness of unbiased LCMS lipid profiling depends on both the mass spectrometer as well as on the separation techniques applied. We studied the robustness of MS profiling using a plasma samples from healthy volunteers including a pooled QC sample which was analysed throughout the batch run. These samples underwent minimal clean-up (protein precipitation) and different fortifications. This paper discusses the potential impact of accelerating high resolution LC separations and the influence on parameter setting in profiling software.
Structure Elucidation of Disulfide Bonds in a Rhamnose-Binding Lectin from Salmon Eggs
| Session: Proteomics: PTM Determination (Method Development) |
Code: MPL |
Poster: 284 |
Liang Zhao1; Ruben T. Almaraz2; Fan Xiang3; Jerry L. Hedrick2; Andreas H. Franz1
1University of the Pacific, Stockton, CA; 2University of California Davis, Davis, CA; 3Shimadzu Biotech, Pleasanton, CA |
Lectins are carbohydrate-binding proteins without glycosidase activity and have well-established roles in cell-cell recognition events. The number of fully-characterized egg lectins of different species has steadily increased over the past decade. The techniques used to analyze disulfide bond patterns have improved dramatically since the development of MALDI and ESI. In combination with enzymatic digestions, chemical modifications, and separation by HPLC, MALDI and ESI have been widely used in protein sequencing and disulfide mapping. The rhamnose-binding lectin from the Chinook salmon Oncorhynchus tshawytscha (SEL24K) has a tandem-repeat sequence of 195 amino acids including 16 cysteines. In this work, the disulfide bond pattern in SEL24K was established unequivocally by applying a multi-enzyme strategy in combination with MALDI-TOF MS/MS analysis.
A Combination of Proteomic and Glycomic Characterization of Prostate-Specific Antigen Using Chip-based MALDI-M
| Session: Carbohydrate/Oligosaccharides |
Code: MPU |
Poster: 490 |
Yan Li1; Lori J. Sokoll1; Brian J. Feild2; Daniel W. Chan1; Hui Zhang1
1Johns Hopkins University, Baltimore, MD; 2Shimadzu Scientific Instruments, Columbia, MD
|
Prostate specific antigen (PSA) is the best-known marker for prostate cancer diagnosis. However, PSA alone is not specific for early stage cancer in the 2-10ng/mL range. Various methods have been evaluated with the goal of improving the specificity of the PSA. One approach is to target different molecular forms of PSA in both amino-acid and carbohydrate portions. It has been shown that a truncated precursor form of PSA ([-2]proPSA) improves the prediction of prostate cancer, and, glycan structures of PSA from prostate cancer sera are different from those from non-cancer sera. This suggests that development of assays to detect both amino-acid and carbohydrate portions of PSA is useful to improve the performance of prostate cancer detection using PSA.
Production and Fragmentation of Negative Ions from Neutral N-Linked Glycans Ionized by MALDI Mass Spectrometry for Rapid Structural Identification
| Session: Carbohydrate/Oligosaccharides |
Code: MPU |
Poster: 498 |
David J. Harvey1; Paula Domann2; Daniel Spencer3; Ian Edwards4; Rachel L. Martin4
1University of Oxford, Oxford, UK; 2LGC Ltd., Teddington, UK; 3Ludger Ltd, Abingdon, UK; 4Shimadzu Biotech, Manchester, UK |
Fragmentation spectra of negative ions from neutral carbohydrates, as seen by CID following electrospray ionization, contain much more structural information than corresponding spectra of positive ions. However, neutral carbohydrates are reluctant to form negative ions by MALDI except from specific matrices such as nor-harmane ([M-H]- ions) or by forming adducts with chloride or sulfate; none of these ions yield ideal MS or MS/MS spectra. We now report an improved method for producing negative ions by doping common matrices with anions such as nitrate and phosphate. Subsequent fragmentation yields similar diagnostic spectra to those obtained by ESI/CID allowing rapid glycan identification from mixtures, not possible before by MALDI. MSn spectra allowed several fragmentation pathways yielding diagnostic ions to be deduced.
High Versatility and Quantitative Capability at Femtomol Level of the Liquid Matrix 3-aminoquinoline/CHCA in MALDI Mass Spectrometry
| Session: MALDI Sample Preparation |
Code: MPZ |
Poster: 569 |
Yuko Fukuyama; Kaoru Kaneshiro; Kenichi Taniguchi; Sadanori Sekiya; Shinichi Iwamoto; Koichi Tanaka
Shimadzu Corporation, Kyoto, Japan |
3-aminoquinoline/CHCA (3AQ/CHCA) is a liquid matrix, which was reported by Kumar et al. in 1996, for MALDI mass spectrometry. Liquid matrixes have generally been reported to have high quantitative performance due to high sample/ matrix homogeneity, but in many cases their sensitivity is much lower than conventional solid matrixes and their quantitative performance is shown at the pmol level. In this study, we have evaluated the 3AQ/CHCA for MALDI-MS. As a result, the 3AQ/CHCA has the general versatility ionizing several substances, for example peptides, proteins, sugars and synthetic polymers. In addition, we have established the preparation condition for the achievement of the quantification with the sensitivity at fmol level.
MALDI-DITMS/MS for High Mass, High Sensitivity and High Resolution Measurement
| Session: MALDI Tandem MS |
Code: MPZ1 |
Poster: 586 |
Koichi Tanaka; Sadanori Sekiya; Shinichi Iwamoto
Shimadzu Corporation, Kyoto, Japan |
Three Dimensional Quadrupole Ion Traps (3D-QIT) have been used to measure mainly low and middle m/z ions. Li Ding, et. al.[1] developed the Digital Ion Trap (DIT) using a rectangular waveform to trap low to high mass ions by changing the trapping frequency and voltage. High mass detection up to IgG (M.W.: 150kDa) was presented at ASMS2008 MP011. Further improvements were applied and IgM (M.W.:~900kDa) can be detected. Isotopic distribution (m/dm>25,000) can be measured from Cytochrome C singly charged ion, and product ions were generated using conventional CID. [1] J. Mass Spectrom., Vol.39, p471-484 (2004)
Tuesday
The Development of a Summarization and Visualization Method for MSn Information Based on Social Network Analysis
| Session: Bioinformatics II |
Code: TPB |
Poster: 067 |
Shinichi Yamaguchi
Shimadzu Corporation, Kyoto, Japan |
Mass spectrometry is increasingly applied to profiling analysis such as metabolomics or proteomics because of its high sensitivity and high separation performance. To analyze the huge information from a mass spectrometer, various statistical data analysis methods are used, like principle component analysis (PCA), partial least squares (PLS), and hierarchical cluster analysis (HCA). We can acquire two kind of information from a mass spectrometer, one is mass (MS1) spectrum, and the other is MS/MS (MSn) spectrum. To visualize and summarize the MS1 spectra, PCA is the most popular method. And for the information of MS/MS spectra, HCA is a major method used to classify them.
Principal Component Analysis (PCA) Applied to MALDI-MS Images, Unsupervised Data Interrogation Directing Peptide Selection from Trypsin-digested Tissue Sections
| Session: Imaging MS: Instrumentation and Method Development |
Code: TPG |
Poster: 174 |
Emrys A. Jones1; Adam McMahon1; Alex Henderson1; Herve Boutin1; Emmanuel Raptakis2; Patricia Price1
1University of Manchester, Manchester, UK; 2Kratos Analytical/Shimadzu Biotech, Manchester, UK |
The information-rich data-set obtained from a single tissue section under MALDI-MS imaging means that manual inspection of the data is very time consuming and almost certainly incomplete. Multivariate statistical analysis tools allow data reduction such that the significant signals are highlighted. PCA is used to reduce the dimensionality of protein maps of brain sections from a range of neurological conditions such as stroke and Alzheimer’s Disease, identifying significant regions and proteins that are to be investigated further by conventional proteomic analyses.
The same PCA techniques are used on-tissue following in-situ trypsin digestion, in this instance grouping together co-located peptides from regions of interest, directing peptide mass fingerprinting (PMF) and MS/MS analysis leading to protein identification
Quantitative Evaluation of Sensitivity for Microscopic AP-MALDI-MS Imaging in Direct Tissue Analysis
| Session: Imaging MS: Instrumentation and Method Development |
Code: TPG |
Poster: 189 |
Takahiro Harada1; Kazuteru Takahashi1; Kiyoshi Ogawa1; Yoshikazu Yoshida1; Yuki Sugiura2; Takahiro Hayasaka3; Mitsutoshi Setou3
1Shimadzu Corporation, Soraku-gun, Japan ; 2Tokyo Institute of Technology, Yokohama, Japan; 3Hamamatsu University School of Medicine, Hamamatsu, Japan |
Recently MS imaging became widely used for pharmaceutical research, biomarker discovery, and basic research. In such researches, not only analysis of organs at the macroscopic scale, but also analysis of more detail structures at small scale is required. To meet such needs, we have been developing a novel imaging mass spectrometer including an optical microscope and an AP-MALDI-QIT-TOF system. A spatial resolution of several µm is achieved by minimizing a laser spot size. Especially in high-resolution analysis, sensitivity is an important issue because spatial resolution and sensitivity are generally contradicting. So we examined a sensitivity of our instrument in practical tissue analysis by evaluating how much of analytes is needed in single laser spot to be detected.
A Desorption Corona Beam Ionization Source for Direct Analysis of Samples from Surface
| Session: Direct Ionization (DESI, DART and ASAP) |
Code: TPL |
Poster: 301 |
Wenjian Sun; Xiaohui Yang; Junsheng Zhang; Tao Lin; Li Ding
Shimadzu Research Laboratory, Shanghai, China
|
The current work involves developing a desorption corona beam ionization (DCBI) source for analyzing samples under atmospheric pressure without sample pretreatment. A visible corona beam can be formed from a stream of helium gas at the tip of an adaptor when a high DC voltage was applied. The gas is heated for desorbing the analytes from surface and the desorbed species are ionized by the energized particles in the corona beam. Samples tested include drug tablets, pesticides, explosives, etc. on different substrates. Visibility of the corona beam in the current work greatly facilitates pinpointing the sampling areas and also makes profiling of the sample surfaces possible.
Development of High-throughput Metabolic Profiling Method Using Highly Sensitive MALDI Mass Spectrometry
| Session: Novel Metabolite Identification Techniques |
Code: TPX |
Poster: 579 |
Daisuke Miura1; Yoshinori Fujimura1; Shinichi Yamaguchi2; Hirofumi Tachibana1; Hiroyuki Wariishi1
1Kyushu University, Fukuoka, Japan; 2Shimadzu Corporation, Kyoto, Japan |
To date, LC-MS or CE-MS based analysis is thought to be a conventional strategy for metabolomics. These methods, however, have several disadvantages such as a large sample consumption, long analytical time, complexity of sample preparation and ion suppression (esp. electrospray ionization). MALDI-MS has several advantages for metabolomics because it is highly sensitive, high-throughput and low sample-consuming methods compared with other analytical platforms such as LC-MS or CE-MS. In the present study, we developed the methodology of the rapid and direct analysis of cellular metabolites by MALDI-TOF-MS for high-throughput and non-targeted metabolomic analysis. The data was applied to multivariate analysis, indicating that this technique is an effective high-throughput cellular metabolic profiling system.
Wednesday
Isobaric Mass Tags for Quantitative Multiplexed Imaging of mRNA Distributions by In-situ Hybridisation and MALDI-MS
| Session: Imaging MS: Peptides and Proteins |
Code: WPG |
Poster: 189 |
Emrys A. Jones1; Adam McMahon1; Andrew Thompson2; Emmanuel Raptakis3
1University of Manchester, Manchester, UK; 2Trillion Genomics, Cambridge, UK; 3
Kratos Analytical/Shimadzu Biotech, Manchester, UK |
Photocleavable mass tags for MALDI imaging of tissue sections allows the mapping of bio-molecules that fall outside the capacity of conventional MS imaging due to low concentration or molecular size. By replacing the fluorescent group on antibody- or antisense-probes with a mass spectrometry ‘friendly’ group, MALDI-MS imaging can be used for immunohistochemical and in-situ hybridisation experiments.
The novelty of the approach presented here is in the isobaric nature of the mass tags. By incorporating an easily cleavable linker into the reporter ion, mapping in MS/MS mode provides greater sensitivity and specificity of detection. Additionally, the isobaric tags allow multiplexing and also normalisation and quantification. These advantages are demonstrated on-tissue with the mRNA of the HER-2 gene.
Fast Analysis of 15 Endogenous Estrogens Using Positive Ion Electrospray with Cumulative Multicolumn Sequential Chromatography for Low Femtogram Analysis
| Session: Clinical Chemistry II |
Code: WPJ |
Poster: 271 |
Timothy D. Veenstra1; Xia Xu1; Haleem Isaaq1; King Chan1; Robert Classon2; William A. Hedgepeth2
1SAIC-Frederick, Inc., Frederick, MD; 2Shimadzu Scientific Instruments, Columbia, MD Liverpool, UK |
While a link between estrogens and cancer has been established, the specific metabolites that play key roles in cancer progression are not currently known. We have previously developed an assay for measuring 15 estrogen metabolites in biofluids, however, sample throughput is low. This throughput is a direct result of many of these steroids having identical compositional formulae and thus identical masses, so the separation requirements to resolve these compounds prior to MS analysis are significant. An analytical approach was developed using dansylation combined with Cumulative Multicolumn Sequential Chromatography to incrementally separate combinations of identical mass steroids. Columns were selected to resolve different pairs and combinations of estrogens without allowing recombination by subsequent columns in the series.
Automatic Non-volatile Salt and Ion-pair Reagent Removal System for Impurity Determination in Pharmaceutical Products
| Session: LC/MS Sample Preparation |
Code: WPN |
Poster: 349 |
Satoshi Yamaki; Naoki Hamada; Yoshihiro Hayakawa; Shuzo Maruyama; Junko Iida
Shimadzu Corporation, Kanagawa, Japan |
In the process of pharmaceutical development, elucidation of molecular structures of impurities is an important step for quality assurance. LC-MS is a recognized technology in product characterization and compound verification studies, however, it is often constrained by methods requiring non-volatile buffer solutions that is based on the Pharmacopoeia. Column switching technique can solve this problem, supplying on-line systems for LC-MS by replacing solvents with a pre-concentration function. This system allows us to select the optimal mobile phase for separation of LC and the optimal mobile phase for MS analysis, respectively. In this study, a practical on-line sample preparation system using column switching HPLC with API-QIT/TOFMS has been evaluated for the structure elucidation of trace amount of compounds in drugs.
The Development of a New Algorithm for Empirical Formula Calculations Based on Multiple Molecular Ion Data
| Session: Natural Products |
Code: WPP |
Poster: 376 |
Ichiro Hirano1; Yusuke Inohana1; Yutaro Yamamura1; Norio Mukai1; Michizane Hashimoto2; Neil Loftus3; John Warrander3
1Shimadzu Corporation, Kyoto, Japan; 2Astellas Pharma Inc., Tsukuba, Japan; 3Shimadzu ISS, Manchester, UK |
Mass spectrometry is increasingly applied to empirical formula prediction in a diverse range of research fields. The likelihood of predicting the correct elemental composition from a list of possible candidates is influenced by a number of factors including the initial chemical knowledge which imposes limits on the element list. To help increase the certainty in formula prediction without imposing element list constraints, an algorithm has been developed that takes into account high mass accuracy, isotope pattern matching, chemical rule filtering and fragment ion data (MSn). This algorithm has now been further enhanced by including multiple adduct ions from a single compound, especially protonated and deprotonated ions, to minimize candidate lists and increase the likelihood of formula prediction.
Determination of Digoxin and Digitoxin by Electrospray Ionization Ion Trap Time-of-Flight Tandem Mass Spectrometry
| Session: Natural Products |
Code: WPP |
Poster: 381 |
Kang Ma2; Leren Wan1; Jing Dong1; Hashi Yuki1; Li Hongmei2
1Shimadzu, Shanghai, China; 2National Institute of Metrology of China, Beijing, China |
Digoxin and digitoxin are the most widely prescribed digitalis glycosides which are well known for the treatment of congestive heart failure. However, the preparation of digoxin and digitoxin is limited to the extraction from natural plants of digitalis lanata due to the absence of the chemical synthetic method. Developing synthetic routes needs the structural information of compounds and their reactive intermediates which can be obtained by mass spectrometry (MS). Especially, the electrospray ionization ion trap time-of-flight tandem mass spectrometry (ESI-IT-TOF-MS) is well suitable for the characterization of the multistage-ion of measured compounds owing to its high sensitivity and mass resolution.
Pancreatic Cancer Serum Detection Using A Lectin/Glyco-Antibody Array Method
| Session: Protein, Glycoproteins |
Code: WPU |
Poster: 507 |
Chen Li1; Eugene Zolotarevsky1; Michelle A. Anderson1; Dean E. Brenner1; Diane M. Simeone1; David M. Lubman1; Fan Xiang2
1University of Michigan, Ann Arbor, MI; 2Shimadzu Biotech, Pleasanton, CA |
Pancreatic cancer (PC) is the fourth leading cause of cancer-related mortality in the US. Patients with PC commonly present with advanced disease. Serologic markers with high specificity and sensitivity can be clinically useful in screening high-risk PC populations to identify early resectable lesions. Alteration in glycosylation patterns of serum glycoproteins is known to be associated with malignant transformation. Identifying unique glycosylation patterns in PC may serve as a unique biomarker of the disease.
Enrichment and Identification of Glycoproteins and Glycan Using Nano-scale Chelating Con A Monolithic Capillary
| Session: Protein, Glycoproteins |
Code: WPU |
Poster: 511 |
Shun Feng1; Na Yang1; Subramaniam Pennathur1; Steve Goodison2; David M. Lubman1; Fan Xiang3
11University of Michigan, Ann Arbor, MI; 2University of Florida, Jacksonville, FL; 3Shimadzu Biotech, Pleasanton, CA |
Glycosylation of proteins is one of the most ubiquitous post-translational modifications observed in eukaryotic organisms. Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. In order to improve efficiency and to make the technique applicable to minimal sample material, we have developed a nano-scale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate–co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure.
Accelerating Japanese Green Tea Quality Assessment by Ultra Fast LC-IT-TOF MS Based Profiling Studies Using High Mass Accuracy MSn Analysis
| Session: Metabolite Profiling |
Code: WPX |
Poster: 576 |
Tairo Ogura1; Takushi Yamamoto1; Satoshi Yamaki1; Tatsunari Yoshida1; Hirohisa Mikami1; Rui Kawahara2; Takeshi Bamba2; Eiichiro Fukusaki2
1Shimadzu Corporation, Kyoto, Japan; 2Osaka University, Osaka, Japan |
The application of LC/MS based global profiling studies in metabonomics/metabolomics research has considerable promise in developing new medicines and in identifying markers for disease, it also has a potential impact on food engineering. In this study, a metabolomics based approach was applied to characterize the quality of Japanese green tea leaves using an ultra fast LC coupled to a quadrupole ion trap-TOF MS system (UFLC-IT-TOF). As the quality of tea is affected by a number of factors, including the variety of tea leaf and the manufacturing process itself, the UFLC-IT-TOF system was used to identify potential markers that could account for the variation in quality and grade classification.
Overcoming the Limitations of MALDI-TOF-MS Analysis of Polymers Using GPC-MALDI and a Hybrid Ion Trap Time of Flight MALDI MS
| Session: Polymers |
Code: WPZ |
Poster: 621 |
Brian Feild1; Fan Xiang2; Martin Resch3; Chrys Wesdemiotis4
1Shimadzu Scientific Instruments, Columbia, MD; 2Shimadzu Biotech, Pleasanton, CA; 3Shimadzu Europe, Duisburg, Germany; 4The University of Akron, Akron, OH |
MALDI-TOF analysis of polymers has gained wide acceptance as an analytical method for confirming structural information and generating polymer statistics. Despite the relatively simple sample preparation and fast analysis time, many difficulties still exist for this technique. For example, co-polymers and polymers of high polydispersity index values (PDI > 2) pose challenges for obtaining structural confirmation and reliable statistics respectively. This presentation will illustrate the ability to improve mass coverage by decreasing spectral complexity with the use of an integrated GPC-MALDI system. Additionally, the high resolution ion selection capability and the ability to perform MSn analysis of the hybrid ion trap - time of flight MALDI MS will be utilized for structural analysis.
Comparative Study of Fatty Alcohol Alkoxylate Copolymers Fragmentation Patterns by MALDI-MS/MS Using Low Energy and High Energy CID
| Session: Polymers |
Code: WPZ |
Poster: 622 |
Volker Wulf1; Martin Resch2; Oliver J. Schmitz1; Hans-Willi Kling3; Siegmar Gaeb1; Michaela Wirtz3
1University of Wuppertal, Wuppertal, Germany; 2Shimadzu Europe, Duisburg, Germany; 3
Cognis GmbH, Dusseldorf, Germany |
The class of fatty alcohol alkoxylates describes surfactants that are synthesised by reaction of fatty alcohols with alkoxides like ethylene oxide, propylene oxide and others. Fatty alcohol alkoxylates are used as nonionic surfactants in home and industrial cleaning and washing agents. They have important properties like foam supression, foam control and wetting effects in these products. Furthermore alkoxylates are also relevant in a broad range of chemical industry applications (e.g. in coating and polymerisation additives or agrochemicals), where these serve as dispersal agents and emulsifiers. Here we present a comparative analysis of fatty alcohol alkoxylate copolymers using MALDI-MS/MS with two different CID techniques.
Developing Application Software Using Applied Biosystems Mass Spectrometer and Shimadzu HPLC to Achieve Multiplexing and Direct Instrument Control in Bioanalysis
| Session: Computer Applications |
Code: WPZ3 |
Poster: 693 |
Leimin Fan1; Richard Koeritz3; Tawakol El-Shourbagy2; Huaiqin Wu2
1Abbott Labs, Abbott Park, IL; 2Abbott Laboratories, Abbott Park, IL; 3Shimadzu Scientific Instruments, Columbia, MD |
Multiplexing of LC-MS is the concept of connecting multiple HPLC streams to one mass spectrometer and running them in alternated injection order. The mass spec waiting time of one HPLC run is used to analyze sample introduced by the other HPLC systems. Currently there is no commercial software available in the market to run multiple independent HPLC (autosampler) with single mass spec. The software developed in our lab achieved flexible multiplexing system configuration, independent HPLC setup and method, freedom of single system to multiplexing switch, full sample information tracking and saving in MS data file.
Thursday
Global Profiling Studies in Tumour Bearing Mouse Models Using High Mass Accuracy MSn Analysis
| Session: Metabolomics III |
Code: ThPC |
Poster: 093 |
Lindsay Lai1,2; Ian Wilson2; Robert Wilkinson2; Rajesh Odedra2; Simon Ashton3; Alan Barnes3; Neil Loftus3
1Manchester University, Manchester, UK; 2AstraZeneca, Alderley Park, UK; 3Shimadzu, Manchester, UK
Global metabolite profiling promises immense potential for early diagnosis, therapy monitoring and for understanding the pathogenesis of many diseases. In this study, metabolite profiling has been used to identify potential biomarkers in several mouse cancer models including colorectal, epidermoid carcinoma and non small cell lung cancer cell tumours using high accuracy MSn analysis. Profiling software and multivariate data analysis was used to construct aligned high mass accuracy data arrays and identify metabolite features that show significant change between sample groups.
Isolation of C-Terminal Peptides by Strong Anion-Exchanger from Proteolytic Digests of Fully Amidated Proteins for Mass Spectrometric Sequencing
| Session: Proteomics: Peptide Sequencing |
Code: ThPG |
Poster: 197 |
Takashi Nakazawa1; Mariko Nakagawa1; Hiroki Kuyama3; Eiji Ando2; Osamu Nishimura3; Minoru Yamaguchi2; Susumu Tsunasawa3
1Nara Women's University, Nara, Japan; 2Shimadzu Corporation, Kyoto; 3Institute for Protein Research, Suita, Osaka |
The C-terminal sequencing can be useful for identifying proteins, especially when the N-terminal amino groups are blocked by post-translational modifications. We developed a method for isolating the C-terminal peptide and analyzing the sequence by mass spectrometry. The method consists of the following three steps: (i) exhaustive amidation of the carboxyl groups in a protein, (ii) proteolytic digestion of the modified protein, and (iii) isolation of the C-terminal peptide by HPLC with a SAX column to trap all the peptides other than the C-terminal one, which should solely lack the free carboxyl group(s). We will report the procedures of the method and the results of MALDI mass spectrometric sequencing of the C-terminal peptide thus isolated.
Characterization of Early Maillard Reaction Products Using MALDI Mass Spectrometry
| Session: Proteomics: PTM Determination (Glycosylation and Phosphorylation) |
Code: ThPJ |
Poster: 248 |
Helen Montgomery1; Gerald Stubiger2; Koichi Tanaka3; Omar Belgacem4
11Shimadzu, Koichi Tanaka MS Research Laboratory, Manchester, UK; 2Medical University of Vienna, Vienna, Austria; 3Shimadzu Corporation, Kyoto, Japan; 4Shimadzu Biotech, Manchester, UK |
The Maillard reaction (L.C. Maillard in 1912) happens non-enzymatically between the reducing end of the sugars (C=O) and a reactive NH2 group of a protein. The detailed characterization of the reaction products is of high importance in the field of food processing but also in physiologies and pathologies of human diseases. We report here the reaction of human serum albumin (HSA) and standard peptides with various sugars. The molecular mass of the whole glycated HSA and the total mass of sugars that reacted with the protein were measured. HSA was then digested in order to locate the reaction sites of each sugar. Modified peptides were fragmented using CID experiments in order obtain signatures at the MSMS level.
Classification of Mycotoxin-producing Fusarium Species Based on MALDI-TOF MS Analyses of Their Intact Spores
| Session: Microbial Analysis |
Code: ThPO |
Poster: 384 |
Martina Marchetti-Deschmann1; Wolfgang Winkler1; Jasmin Kemptner1; Emmanuel Raptakis2; Irina S. Druzhinina1; Robert Mach1; Christian P. Kubicek1; Guenter Allmaier1
1Vienna University of Technology, Vienna, Austria; 2Shimadzu Biotech/Kratos Analytical, Manchester, UK |
The differentiation of genetically closely related microorganisms using MALDI-TOF-MS is a task successfully addressed by many research groups but only few of them focus on fungi. However for agriculture, fungi (e.g. Fusarium species) are a problem which has to be dealt with as their corn infestation and mycotoxins production is a subsequent health threat to human. A fast and reliable method to evaluate the pathogenicity of the individual species has to be found. Although fungi morphology significantly differs from bacteria, characteristic peptide/protein pattern desorbed directly from intact spore surfaces provide good mass spectral information to differentiate between Fusarium species. Statistical analysis of the MS data set helps to achieve the final aim to develop a database for fast identification.
Comprehensive Pathogen Identification Using MALDI TOF Coupled to Statistic Patented Procedure
| Session: Microbial Analysis |
Code: ThPO |
Poster: 386 |
Fan Xiang1; Joachim Dyck2
1Shimadzu Biotech, Pleasanton, CA; 2AnagnosTec, Potsdam, Germany |
Automated identification systems are widely-used in medium-to-high-throughput clinical microbiology laboratories. However, such systems are relatively slow because they depend on bacterial growth and metabolic activity. Bacterial identification by mass spectrometry provides a promising way to accelerate pathogen identification, since it can be performed in a few minutes from small samples. In this study we compared the performance of MALDI-TOF MS coupled to SARAMIS (Spectral ARchiving And Microbial Identification System), a patented procedure for statistic evaluation and weighting of data, with established methods (VITEK2/API, BioMérieux) in the clinical microbiology routine diagnostics.
Mass Spectrometric Analysis of Proteolytic Products from MBP-Containing Heterologous Proteins Expressed in Pichia pastoris
| Session: Proteins: Recombinant |
Code: ThPU |
Poster: 547 |
Zhiguo Li1; Wilson Leung1; Joan Lin-Cereghino1; Geoffrey Lin-Cereghino1; Fan Xiang2; Andreas Franz1
1University of the Pacific, Stockton, CA; 2Shimadzu Biotech, Pleasanton, CA |
Pichia pastoris is a methylotrophic yeast that has been genetically engineered to express heterologous proteins for important industrial, pharmaceutical and basic research purposes. Over the past 20 years, more than 700 proteins from bacteria to humans have been produced in this yeast. Maltose Binding Protein (MBP) has been utilized as a translational fusion partner to improve the expression of foreign proteins made in prokaryotic E. coli. Although the mechanism of its action is not completely understood, MBP has been shown to increase the solubility of intracellular proteins as well as improve protein secretion. Here, we explored whether MBP would have the same effect in eukaryotic Pichia pastoris.
A New Multi-turn Time-of-Flight Mass Spectrometer with High Resolving Powers above One Million
| Session: Instrumentation: TOF |
Code: ThPY |
Poster: 625 |
Osamu Furuhashi1; Kengo Takeshita1; Hideaki Izumi1; Shinichi Yamaguchi1; Masaru Nishiguchi1; Hiroki Sakae1; Yoshihiro Ueno1; Kiyoshi Ogawa1; Yoshikazu Yoshida1; Michisato Toyoda2; Mitsutoshi Setou3
1Shimadzu Corporation, Kyoto, Japan; 2Osaka University, Toyonaka, Osaka, Japan; 3Hamamatsu University School of Medicine, Hamamatsu, Japan |
Mass spectrometry plays an important role in life science and there are still increasing needs for higher performances. Among various methods, multi-turn time-of-flight (TOF) mass spectrometer, in which ions repeatedly travel in a closed orbit, can improve mass resolving powers with retaining high throughput and small size. Last year, we proposed a high-performance multi-turn TOF mass spectrometer based on a new design concept; the time focusing and the stability of trajectory in the phase space have been required. Both high resolving powers and high ion transmission were predicted for the optical system. Based on this study, we have newly constructed a multi-turn TOF mass spectrometer. Some remarkable results have been obtained experimentally and will be given in this presentation.
Co-polymer Characterization Using Automated On-line SEC-Pyrolysis GCMS
| Session: High Throughput Analysis/Robotics |
Code: ThPZ1 |
Poster: 681 |
Junko Iida1; Erwin Kaal2,3; Hans-Gerd Janssen2,4
1Shimadzu Europa GmbH, Duisburg, Germany; 2Van´t Hoff Institute for Molecular Sciences, Amsterdam, The Netherlands; 3Atas GL International, Veldhoven, The Netherlands; 4Unilever Research and Development, Vlaardingen, The Netherlands |
The properties of a polymeric material depend on the chemical composition of a polymer as well as the molecurlar weight distribution. In this study a stop flow operation of SEC was coupled on-line to a GCMS instrument equipped with a temperature programmable injector, capable of high temperatures. In such a way solvent elimination and subsequent pyrolysis can be performed.
Fast Analysis of Vitamins in Dietary Supplements Using LCMS
| Session: High Throughput Analysis/Robotics |
Code: ThPZ1 |
Poster: 683 |
Masatoshi Takahashi; William A. Hedgepeth; Yuhui Wang
Shimadzu Scientific Instruments, Columbia, MD |
Commercially available sub-2µm or sub- 3µm particle columns have been used to shorten analytical time in a wide range of fields. The sub-3µm columns, such as a 2.2µm particle column, give similar performance to sub-2µm column with much lower pressures, allowing the column to be accommodated by conventional systems. With the improvement of separation speeds, it has become important to acquire MS data more quickly and accurately by using high-speed scanning and polarity switching. This paper describes the application of a high speed analysis of vitamins in dietary supplements, which have received a lot of attention with the implementation of new cGMPregulations, by ultra fast LC-hyphenated MS spectrometer.
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