Group: Danaher
Catalog excerpts
ANSWERS FOR SCIENCE. KNOWLEDGE FOR LIFE. SelexION™ Technology for Lipid Analysis: Pushing the Boundaries of Lipidomics Baljit Ubhi, Ph.D ASMS Baltimore, June 2014
Open the catalog to page 1Merged lipidomic dataMultiple data analysis tools Unraveling the Multiple lipidomic platforms identification of Biomarkers Liquid-liquid extraction Data Processing Interpretation & Reporting Lipidomic Analysis flLJR LipidView ■ btimlW LJfad Oianclart Stahlman, M. et al J Chromatogr B Analyt Technol Biomed Life Sci. 2009
Open the catalog to page 2Comprised of Multiple, Distinct Structural Lipid Classes Fatty Acids Steroids Lipid Terpenes Vitamins Mono-, Di- and Waxes Sphingolipids Triglycerides Glycerophospholipids I Sphingomyelins Ceramides Cerebrosides Gangliosides Ether Phospholipids Activating Plasmalogens Diacyl-Linked Phospholipids Oxidized Phospholipids Halogenated Lipids Lipids play an essential role in human physiology: • Metabolic homeostasis • Cell signaling • Cell and organelle structure And disease: • Inflammation • Cancer • Cardiovascular disease • Diabetes • Inflammatory bowel disease • Neurological diseases
Open the catalog to page 3• Lipidomic spectra are incredibly complex • MS/MS spectra generated on precursors in zones of isobaric overlap will contain product ions from other isobaric species
Open the catalog to page 4SelexION™ Technology Ion Mobility Device Components 1. Orifice Plate • Robust, easy-to-install, hardware components: ‒ ‒ ‒ ‒ 5 No tools required No cables No need to break vacuum Installation in about 2 minutes
Open the catalog to page 5How Does DMS Separate Ions? Differential Mobility Spectrometry (DMS) is the term used for planar geometry Gas/Modifier flow Separation waveform (SV): radially displaces ions towards one or the other electrode, depending upon high and low mobility characteristics 6 Compensation voltage (COV): restores the trajectory for a given ion or range of ions to allow them to transmit through the DMS device and enter the mass spectrometer
Open the catalog to page 6SelexIONTM Technology Separates Phospholipid Classes Experiment: MRM Scan of 6 Phospholipid Standards with COV Ramp • Using DMS alone, a mixture of lipids can be separated into it’s individual components • Baseline separation can be achieved, completely abrogating isobaric interference Proof of concept: DMS separates simple lipid mixtures Implications: “Cleaner” quantitative data using mrm scan modes (esp., MRM HR); Improved qualitative analysis 7
Open the catalog to page 7Effects of DMS on Complex Lipid Analysis Can DMS help resolve lipids from a complex, biological lipid mixture? Experiment 1: EMS Scan of Bovine Heart Extract; No DMS 723.70 7e6 6e6 5e6 Experiment 2: EMS Scan of Bovine Heart Extract; DMS with COV Ramp 8.3 -1.5
Open the catalog to page 8An individual phospholipid subclass can be extracted from an EMS scan based on its differential COV • Simplifies LipidView™ Software searching • Increases confidence in lipid identification
Open the catalog to page 9Relationship Between Dipole Moment and COV 8 4.5 5 Model headgroup dipole moment (D) Isopropanol as a chemical modifier in DMS 10
Open the catalog to page 10Lipid Analysis by AB SCIEX TripleTOF® 5600+ System • Mass Range ‒ Q1 selection – 5 - 1250 m/z ‒ TOF MS – 5 – 40 000 m/z • Speed ‒ 10 ms minimum accumulation times ‒ Up to 100 MS/MS scans per second in IDA • Resolution ‒ High resolution mode ~30,000 ‒ High sensitivity mode ~15,000 • Mass Accuracy ‒ 1-2 ppm RMS (external calibration)
Open the catalog to page 11TOF MS Analysis of Lung Lipid Extract Using DMS Phospholipids; Cardiolipin Fatty Acids Experiment: TOF MS with COV ramp in negative ion mode; Infusion of 100 μg/mL lung lipid extract 12
Open the catalog to page 12Experiment: TOF MS with COV ramp in negative ion mode; Infusion of 100 pg/mL lung lipid extract > XIC from DMS Expenmenls.wit«sample 12) - S«mple5X-2 DMSon COVromp DR=44 NtG-2.5a
Open the catalog to page 13DMS Resolves Fatty Acid Cis/Trans Isomers Experiment: TOF MS with COV ramp in negative ion mode; Infusion of 100 μg/mL lung lipid extract Resolution of cis/trans isomers usually requires chromatographic separation prior to analysis; DMS can remove this need, increasing the speed of FA analysis
Open the catalog to page 1411C trom UMb bxpenments.wm (sample 14) - bample pa-4 UMb on CUV ramp UK=44 NbG 4.0 acc, -1 Uh Mb i.lUU - 4UUU) Spectrum from DMS Experiments.wiff (sample 12) - Sample 5X-2 QMS on COV ramp DR=44 NEG “ 2.5 acc, -TOF MS (100 - 2000) from 13.658 to 13.827 min
Open the catalog to page 15f£J A ♦ ’ % A 1a w ’ «-tA I QSSlStf Spectrum from DMS Experiments.wiff (sample 6) - Sample 5X-2 DMS on COV ramp DR=42.5, +TOF MS (100 - 2000) from 3.517 to 3.740 min COV = 0.4 to 1.7 V
Open the catalog to page 18Resolution of Triglycerides from Complex Mixtures of Lipids COV for TAG
Open the catalog to page 19SLICE: Structural Lipidomics Investigations using Chemical Effects • Molecules are separated in CV space prior to entering the MS • Chemical modifiers enhance compound resolution DMS-Separated Precursors • Minimizes isobaric overlap MS/MS of Selected Precursors
Open the catalog to page 20Confident Identification of Isobaric Compounds Experiment: DMS MS/MSALL (NEG) of bovine heart extract © 2014 AB SCIEX Duchoslav E., Campbell, J. L., Baker, P.R.S., Patterson B.T., ASMS 2013, ThP 28-586
Open the catalog to page 21Comparison of lipid profiles obtained with and without DMS separation * Species with 3 or more chains (TAG, CL) are represented as sum of all permutations
Open the catalog to page 22SelexION™ Technology for Lipidomic Analysis Effective lipid resolution without a need for complicated chromatographic strategies • SelexION™ Technology isolates lipid classes prior to MS/MS analysis, which reduces isobaric and isotope overlap and enables accurate identification and quantitation • Lower LOQ/LOD for targeted lipidomics; reduction in matrix interference
Open the catalog to page 23Zora Biosciences •Kim Ekroos •Tuulia Sylvanne Wollongong University • Todd Mitchell • Steven Blanksby • Simon Brown • Al Maccarone UCSD • Ed Dennis • Oswald Quehenberger • Paul Norris • Aaron Armando Eli Lilly • Phil Sanders AB SCIEX • Paul Baker • Larry Campbell • Michael Jarvis • Eva Duchoslav • Brigitte Simons • Christie Hunter • Baljit Ubhi Avanti Polar Lipids LipidMAPS.org
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