C10G-E065A Fundamental Guide to Liquid Chromatography Mass Spectrometry (LCMS) MILESTONES FOR LIQUID CHROMATOGRAPHY AND MASS SPECTROMETRY Shimadzu Corporation focus on analytical and measuring instruments, Genzo Shimadzu Sr. founds medical systems, aircraft equipment and industrial machinery. Together with 1875 our partners around the world, we diligently strive to be a company that Shimadzu Corporation meets the challenges faced by society and develop products which contribute to the growth of industries and the enrichment of peoples’ lives. WORLD’S 1ST MALDI-MS 1987 LAMS-50K LCMS-2010 NOBEL PRIZE IN CHEMISTRY AWARDED TO KOICHI TANAKA 2002 AXIMA-CFR LCMS-2010A AXIMA-QIT Koichi Tanaka Mass Spectrometry Research 2003 Laboratory established World-Class General Purpose 2004 World’s 1st Ion trap TOF LCMS LCMS-IT-TOF 2006 Prominence UFLCXR Opening up new worlds and expanding possibilities LAUNCH OF UFMS TECHNOLOGY 2008 through ultra-fast liquid chromatography MALD-TOF MS models Shimadzu’s revolutionary technology Acquire mass spectra directly from bringing next-generation analysis crude samples performance LCMS-2020 © Shimadzu Corporation 2010 Delivers superior performance across a wider First Edition: October, 2018 Minimized cross talk application range with flexibility and reliability LCMS-8030 with UFsweeper™ Updated on: March, 2019 MALDI-7090 Higher sensitivity with UF-Lens™ Ultimate performance in identification and structural LCMS-8040 Improved CID efficiency with 2013 characterization of biomolecules UFsweeperII™ 2014 LCMS-8050 Integrated LC systems that provide high speed analysis, simplified For Research Use Only. Not for use in diagnostic procedures. Achieved UFswitching™, Next-level method transfer, minimized environmental impact and easy maintenance UFscanning™ and UF-MRM™ quantitative Provides high resolution This publication may contain references to products that are not available in your country. Please contact us to check the detection with morphological imaging overlaid availability of these products in your country. UF-Qarray™ with molecular distribution images The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Unified Chromatography (UC) 2015 UHPLC/SFC Switching System Shimadzu. LCMS-8060 World's 1st fully automated system that combines Company names, products/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation, its on-line SFE and SFC in a single flow path. subsidiaries or its affiliates, whether or not they are used with trademark symbol “TM” or “®”. Third-party trademarks and trade names may be used in this publication to refer to either the entities or their products/services, whether World’s 1st Automated LCMS or not they are used with trademark symbol “TM” or “®”. LCMS-8045 2016 Module for Clinical Applications CLAM-2000 Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own. Cutting-edge UFsweeperIII™, DPiMS-2020 Quick and easy MS analysis The information contained herein is provided to you “as is” without warranty of any kind including without limitation warranties as to UF-grating™ and UF-FlightTube™ without sample preparation its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating allows high-speed data acquisition to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, MALDI-8020 and subject to change without notice. for high-throughput analysis LCMS-9030 Nexera Mikros Quadruopole Time-of-Flight 2018 With the added UF-Link, this low flow LCMS system Liquid Chromatography Mass Spectrometer improves operability and enables high-sensitivity MILESTONES FOR LIQUID CHROMATOGRAPHY AND MASS SPECTROMETRY Shimadzu Corporation focus on analytical and measuring instruments, Genzo Shimadzu Sr. founds medical systems, aircraft equipment and industrial machinery. Together with 1875 our partners around the world, we diligently strive to be a company that Shimadzu Corporation meets the challenges faced by society and develop products which contribute to the growth of industries and the enrichment of peoples’ lives. WORLD’S 1ST MALDI-MS 1987 LAMS-50K LCMS-2010 NOBEL PRIZE IN CHEMISTRY AWARDED TO KOICHI TANAKA 2002 AXIMA-CFR LCMS-2010A AXIMA-QIT Koichi Tanaka Mass Spectrometry Research 2003 Laboratory established World-Class General Purpose 2004 World’s 1st Ion trap TOF LCMS LCMS-IT-TOF 2006 Prominence UFLCXR Opening up new worlds and expanding possibilities LAUNCH OF UFMS TECHNOLOGY 2008 through ultra-fast liquid chromatography MALD-TOF MS models Shimadzu’s revolutionary technology Acquire mass spectra directly from bringing next-generation analysis crude samples performance LCMS-2020 2010 Delivers superior performance across a wider Minimized cross talk application range with flexibility and reliability LCMS-8030 with UFsweeper™ MALDI-7090 Higher sensitivity with UF-Lens™ Ultimate performance in identification and structural LCMS-8040 Improved CID efficiency with 2013 characterization of biomolecules UFsweeperII™ 2014 LCMS-8050 Integrated LC systems that provide high speed analysis, simplified Achieved UFswitching™, Next-level method transfer, minimized environmental impact and easy maintenance UFscanning™ and UF-MRM™ quantitative Provides high resolution detection with morphological imaging overlaid UF-Qarray™ with molecular distribution images Unified Chromatography (UC) 2015 UHPLC/SFC Switching System LCMS-8060 World's 1st fully automated system that combines on-line SFE and SFC in a single flow path. World’s 1st Automated LCMS LCMS-8045 2016 Module for Clinical Applications CLAM-2000 Cutting-edge UFsweeperIII™, DPiMS-2020 Quick and easy MS analysis UF-grating™ and UF-FlightTube™ without sample preparation allows high-speed data acquisition for high-throughput analysis MALDI-8020 LCMS-9030 Nexera Mikros Quadruopole Time-of-Flight 2018 With the added UF-Link, this low flow LCMS system Liquid Chromatography Mass Spectrometer improves operability and enables high-sensitivity Foreword Hirano Ichiro LCMS Product Manager We are very happy to deliver you our first LCMS Primer which has very well compiled the basic principles and theory of mass spectrometry. It also describes the history of development of various innovative technologies to enhance the performance of our LC-MS/MS systems. It serves both as a textbook and a practical application guide, regardless of the scientific background and working field of the readers. All market figures indicate that Shimadzu is the fastest-growing mass spectrometry company in the last decade. Since the product launch of our first triple quadrupole LC-MS/MS, LCMS-8030 in September 2010, we have been continuously rolling out the new LC-MS/MS systems to keep up to date with increasing needs. This year (2018), we are celebrating our landmarking launch of the LCMS-9030, our first ever Q-TOF instrument. Given these achievements, it is now our obligation and responsibility to deliver a primer to increase general understanding of the scientific foundation that we stand upon, while also pointing to the future of the LCMS that we are to realize. I would like to express my gratitude for your eagerness to learn more about LCMS and taking time to read this primer. I sincerely hope that it satisfies your enthusiasm and gets you prepared for the new era of LCMS. List of Contents Chapter 1 Introduction to LCMS 4 Fundamentals of LC, MS and LCMS 5 Comparison of LCMS and other techniques 9 Applications of LCMS 11 Chapter 2 Integrating LC and MS 12 Basic instrumentation of LCMS 13 Interfaces for LCMS 15 Electrospray Ionization (ESI) 16 Atmospheric Pressure Chemical Ionization (APCI) 18 Atmospheric Pressure Photoionization (APPI) 18 Overview of Atmospheric Pressure Ionization (API) 21 Mobile phases compatible for LCMS 24 Chapter 3 Principles of MS 26 Introduction to mass analyzers 27 Magnetic sector MS 28 Quadrupole MS 30 Time-of-Flight (TOF) MS 33 Ion Trap (IT) MS 37 Comparison of mass analyzers 39 Chapter 4 Introduction to tandem MS systems 41 Core principles 42 Types of MS/MS systems and their key characteristics 46 Triple Quadrupole (TQ) MS 47 Quadrupole Time-of-Flight (Q-TOF) MS 47 Multistage MS (MSn) 50 Ion Trap Time-of-Flight (IT-TOF) MS 50 Comparison of MS/MS systems 51 Chapter 5 Challenges and development of LCMS 53 Analytical challenges and current limitations of LCMS 54 Key developments in Shimadzu’s LCMS 56 Ultra-Fast Mass Spectrometry (UFMS) 56 Clinical Laboratory Automation Module (CLAM) 58 Future trends of LCMS 59 Shimadzu’s Comprehensive LC and MS Solutions 60 Abbreviations APCI Atmospheric Pressure Chemical Ionization API Atmospheric Pressure Ionization APPI Atmospheric Pressure Photoionization CID Collision-induced Dissociation CLAM Clinical Laboratory Automation Module DUIS Dual Ion Source EI Electron Ionization ESI Electrospray Ionization GC Gas Chromatography GCMS Gas Chromatography Mass Spectrometry HILIC Hydrophilic Interaction Liquid Chromatography HPLC High-Performance Liquid Chromatography ICP-MS Inductively Coupled Plasma Mass Spectrometry IEC Ion Exchange Chromatography IT Ion Trap IT-TOF Ion Trap Time-of-Flight LC Liquid Chromatography LCMS Liquid Chromatography Mass Spectrometry MALDI Matrix-Assisted Laser Desorption/Ionization MALDI-TOF Matrix-Assisted Laser Desorption/Ionization Time-of-Flight MRM Multiple Reaction Monitoring MS Mass Spectrometry