METABOLOMICS. Biomarker and Omics Solutions FOR METABOLOMICS

Transcription

1 METABOLOMICS Biomarker and Omics Solutions FOR METABOLOMICS

2 The metabolome consists of the entirety of small molecules consumed or created in the metabolic processes that sustain life. Metabolomics is the systematic study of these processes and the molecules involved. Complimentary to genomics, transcriptomics, and proteomics, metabolomics can provide insights into cellular processes and the health and inner-workings of biological systems. At AB SCIEX, we develop tools to help you discover more in your metabolomics research. With innovative hardware, software, and workflows developed specifically for delving deeper into the metabolome, we help you further understand and generate new scientific hypotheses. Start with AB SCIEX, end with new discoveries. Intelligent workflows combined with powerful technology Our industry leading hardware and application-focused workflows deliver answers to your most pressing metabolomics questions. Discover and Screen TripleTOF 5600 system Track and Quantify QTRAP 4500 system 4.0e6 3.5e6 3.0e e6 2.0e6 1.5e6 1.0e6 5.0e5 e e5 100% * (1) * % 3e5 * % * e5 40% * e (1) 20% * * * (1) * e0 0% Mass/charge, Da % Single injection (quant and qual) itraq and mtraq reagent kits Mass/charge, Da MarkerView Non-targeted metabolomics global profiling MRM assay PeakView Targeted metabolomics pathway screening LipidView Lipid profiling With one simple acquisition and any one of three specially designed processing pathways, the TripleTOF 5600 System provides all of the data you need to discover and screen for a wide variety of metabolites. 1.4e5 1.3e5 1.1e5 9.0e4 7.0e MultiQuant Targeted metabolite quantification QTRAP systems help you track and quantify known metabolites of interest in a rigorous hypothesis-driven mode.

3 A pathway to success Metabolomics research occurs along a continuous pipeline. Typically, it begins with a small number of samples to test hypotheses and identify and characterize key metabolites. These metabolites are then verified and validated in greater numbers of samples to account for biological variation. Finally, assays are developed for routine quantitation of the metabolites. Metabolomics research pipeline SCREEN AND DISCOVER QUANTIFY VALIDATE Solutions for metabolomics research Advion TriVersa NanoMate Eksigent ekspert microlc 200 System Eksigent ekspert ultralc 100-XL LC HARDWARE COMPELLING HARDWARE The TripleTOF 5600 system and QTRAP 5500 system along with Eksigent and Advion technologies provide a solid foundation for delivering the highest quality data in a user-friendly and efficient package. MS HARDWARE PIONEERING SOFTWARE Identify, characterize, and screen for hundreds of AB SCIEX TripleTOF 5600 System AB SCIEX QTRAP Systems AB SCIEX QTRAP Systems SOFTWARE metabolites using PeakView, MarkerView and LipidView software. Then accurately track and quantify those metabolites across hundreds of samples using MultiQuant Software. MarkerView PeakView LipidView MultiQuant SCIEX ichemistry SOLUTIONS Reagents and consumables custom-designed to boost the performance of your mass spectrometer and improve sensitivity, productivity, and data precision. itraq, mtraq and atraq labeling reagents KITS Get up and running fast with kits and reagents designed and optimized to track and quantify specific metabolites of interest.

4 One data file. Three processing strategies. Unknown compound identification With the TripleTOF 5600 system and one sample injection you can acquire one data file using our profile workflow, and process that data in three unique and very powerful processing strategies for qualitative and quantitative information: 1 ] Non-targeted metabolomics - Global profiling 2 ] Targeted metabolomics - Pathway screening 3 ] Lipid profiling 1] Non-targeted metabolomics - Global profiling Global metabolite profiling is used to identify interesting metabolites and potential biomarkers in a non-targeted fashion. For global metabolite profiling LC/MS data was acquired across multiple samples using the profile workflow and opened into MarkerView Software for simultaneous feature finding, alignment, and statistical analysis to highlight metabolites of interest. Find the formula: Formula Finder in PeakView Software calculates the elemental composition / formula using precursor and fragment masses. Structural elucidation TOF MS * (1) * (1) For interesting peaks, query the MS and MS/MS data using Formula Finder and TOF MS/MS the Structural Elucidation tool within PeakView Software. e0 * (1) N-acetyl glutamic acid Highlighting metabolites of interest: The PCA scores plot (left) shows samples that group together due to common variables. The PCA loadings plot with principal component variable grouping (PCVG) shows correlation between common and significant variables. Clarify the structure: Structural Elucidation in PeakView Software matches fragment ions from the MS/MS data to the compound structure.

5 2] Targeted pathway screening Sphingomyelin 3] Lipid profiling Targeted pathway screening is a powerful workflow for highlighting up/down regulation of metabolites in biochemical pathways such as glycolysis and TCA cycle. Known biochemical pathways can be screened by analyzing data with XIC Manager within PeakView Software. A pre-defined target list is uploaded to the XIC Manager (e.g., glycolysis pathway metabolites, TCA pathway metabolites, etc.) and compounds are extracted and identified using accurate mass MS and retention time data. Peaks of interest are then confirmed within PeakView Software by performing a library search of the MS/MS data. AdoHcy DNA methylation other methylations Tetrahydrofolate methyl-thf SAM Methionine Vit. B12 Homocysteine ceramide Phosphatidylcholine PtdEtn Betaine CDP-Choline Phosphorylcholine Choline Acetylcholine For in-depth lipid analysis, LipidView Software enables differential profiling of specific lipid species by identifying and confirming lipids with it s built-in library feature. Data files are opened in the software and searched using a database containing over 25,000 lipid species, representing 44 lipid classes and more than 600 characteristic lipid fragments. methyl-groups for emthyl-thf Sarcosine 1.5e5 Betaine: /0.512 Betaine: /0.616 Methionine: 15583/0.618 Cystathionine: /3.179 N-methyl Nictotinamide: /3.783 e0 Homocysteine: /1.278 Taurine: /2.500 Cystathionine: / Lipid identification: LipidView Software identifies lipid species and displays the relative amounts of each lipid species across multiple sample types. Here, 5 samples are compared with a control for 83 different lipid species from positive-ion mode data. 3e4 2e4 1e4 0e0 * (1) TOF MS * % * * (1) * *12654 (1) * % % TOF MS Once lipids are identified, the data files can be exported into MarkerView Software for differential profiling using statistical analysis tools. Extract, list, and confirm: The XIC Manager showing screening and library search results of metabolites in the homocysteine pathway. ID and confirmation use four important search features: mass accuracy, retention time, isotope pattern and library hit with purity score.

6 Targeted quantitation Once metabolites of interest are found, the next step is to monitor and quantitate them across many samples and experiments. Using an MRM workflow on a QTRAP System, metabolites of interest are monitored in a targeted fashion, but with the capacity to rigorously quantify 1000 s of metabolites from a single injection. Design your own method, or use a reagent kit and standards to monitor a set of pre-selected compounds such as atraq for amino acids. The workhorse for quantitative data processing is MultiQuant Software. No other software program delivers the algorithmic power to definitively quantitate at the lowest levels. MultiQuant Software features a fast and userfriendly interface with linked panes, predefined advanced queries for faster data review, saturation correction for increased linear range, and the SignalFinder Integration Algorithm for more consistent peak integration and less user intervention. Dynamic linking: MultiQuant Software features a user-friendly interface with dynamically linked panes. Selecting results in the top pane automatically displays corresponding peak integrations and calibration curves. 1.4e5 1.3e5 1.1e5 9.0e4 7.0e e4 Sar bala Ala GABA lle Leu baib Abu 9.0e4 Nle Forty-five amino acids are quantified a simple 18-minute MRM experiment. The sample is labeled with the atraq reagent and then mixed with internal standards labeled with the same tag containing a different number of isotopes. The same amino acid labeled with the different tags cannot be distinguished by HPLC since they have identical retention times. In the MS/MS mode the tagged fragment yields reporter ions that have been encoded with isotopes to yield different masses. The unique MRM transitions, made up of the labeled mass of the amino acid and the reporter ion mass, are used to determine the amino acid amounts in the sample.

7 You invest in our technology. We invest in your success. As the world leader in mass spectrometry, AB SCIEX creates solutions that are backed by the industry s most extensive service and support organization. With a network of service professionals, experienced compliance specialists, and over 150 PhD application scientists worldwide, we are dedicated to supporting your technical needs and helping you get the most out of your AB SCIEX systems. AB SCIEX service professionals are recognized as the most highly qualified in the industry. They are certified on our instrument platforms through a rigorous 4-step certification program. This award-winning program helps to ensure that you receive the most efficient, highest-quality, and most up-to-date service available for AB SCIEX products and technology. Choose from flexible service plans and a variety of services for the right level of support for your laboratory s needs and budget. Our customer support network is available to provide expert assistance in the use and application of AB SCIEX products through a comprehensive range of services, including application support, technical service, and training. Whether you access our service and support team by phone, , on-site visits, or through our innovative remote monitoring technology, you can be confident that the AB SCIEX organization will be there for you. For more information, visit For Research Use Only. Not for use in diagnostic procedures AB SCIEX. The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners. AB SCIEX is being used under license /2012 Headquarters 500 Old Connecticut Path Framingham, MA USA Phone International Sales For our office locations please call the division headquarters or refer to our website at