DETERMINATION OF POLYMER STRUCTURES, SEQUENCES, AND ARCHITECTURES BY MULTIDIMENSIONAL MASS SPECTROMETRY A Dissertation Presented to The Graduate Faculty of The University of Akron In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy Aleer Manyuon Yol August, 2013 DETERMINATION OF POLYMER STRUCTURES, SEQUENCES, AND ARCHITECTURES BY MULTIDIMENSIONAL MASS SPECTROMETRY Aleer Manyuon Yol Dissertation Approved: Accepted: ________________________________ _________________________________ Advisor Department Chair Dr. Chrys Wesdemiotis Dr. Michael J. Taschner ________________________________ _________________________________ Committee Member Dean of the College Dr. Leah P. Shriver Dr. Chand K. Midha ________________________________ _________________________________ Committee Member Dean of the Graduate School Dr. Claire A. Tessier Dr. George R. Newkome ________________________________ _________________________________ Committee Member Date Dr. Wiley J. Youngs ________________________________ Committee Member Dr. Yu Zhu ii ABSTRACT The matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-ToF/ToF MS) characteristics of different polystyrenes and polybutadienes are discussed in this dissertation. The compounds examined include linear, cyclic, in-chain substituted, and star-branched polymers as well as copolymers of styrene and either para-dimethylsilyl styrene (p-DMSS) or meta-dimethylsilyl styrene (m-DMSS). Chapter IV describes the differentiation of cyclic and linear polymers by 2D- mass spectrometry. The silverated quasimolecular ions from cyclic and linear polystyrenes and polybutadienes, formed by MALDI, give rise to significantly different fragmentation patterns in tandem mass spectrometry (MS2) experiments. With both architectures, fragmentation starts with homolytic cleavage at the weakest bond, usually a C–C bond, to generate two radicals. From linear structures, the separated radicals depolymerize extensively by monomer losses and backbiting rearrangements, leading to low-mass radical ions and much less abundant medium- and high-mass closed-shell fragments that contain one of the original end groups, along with internal fragments. With cyclic structures, depolymerization is less efficient, as it can readily be terminated by intramolecular H-atom transfer between the still interconnected radical sites (disproportionation). These differences in fragmentation reactivity result in substantially different fragment ion distributions in the MS2 spectra. Simple inspection of the relative iii intensities of low- vs. high-mass fragments permits conclusive determination of the macromolecular architecture, while full spectral interpretation reveals the individual end groups of the linear polymers or the identity of the linker used to form the cyclic polymer. Chapter V presents the first sequence analysis of styrenic copolymers by tandem MS. Copolymers of para-dimethylsilyl styrene (p-DMSS) or m-DMSS with styrene were prepared by living anionic polymerization. The MALDI-MS2 results for p-DMSS indicate that a block copolymer is formed, with the para-substituted styrene incorporated near the initiator. On the other hand, the MS2 results of m-DMSS reveal that a random copolymer is formed, consistent with comparable reactivities for m-DMSS and styrene. These findings suggest that p-DMSS is more reactive than m-DMSS. The single-stage (1D) MALDI-MS results further show that linear and 2-armed architectures are formed with both the m-DMSS and the p-DMSS comonomers. The last Chapter, VI, focuses on the differentiation of linear in-chain substituted, cyclic, and star-branched polystyrene (PS) by tandem mass spectrometry. The in-chain functionalized PS gives a MS2 fragmentation pattern that is different from the one observed for cyclic PS with two linker units and, again, with a simple inspection of the tandem mass spectra, these architectures can easily be distinguished. The four-arm star- branched polymer investigated mainly breaks down by losing arms under MALDI-MS2 conditions. Overall, this dissertation documents the usefulness of combined 1D and 2D mass spectrometry experiments for the identification of polymer substituents and their location, for distinguishing polymer architectures, and for determining copolymer sequences. iv The results presented in this dissertation have been published or are pending for publication in the following journals. 1. Quirk, R. P.; Wang, S-F.; Foster, M. D.; Wesdemiotis, C.; Yol, A. M. “Synthesis of Cyclic Polystyrenes Using Living Anionic Polymerization and Metathesis Ring-Closure” Macromolecules 2011, 44, 7538-7545. 2. Liu, B.; Quirk, R. P.; Wesdemiotis, C.; Yol, A. M.; Foster, M. D. “Precision Synthesis of ω-Branch, End-Functionalized Comb Polystyrenes Using Living Anionic Polymerization and Thiol-Ene “Click” Chemistry” Macromolecules 2012, 45, 9233-9242. 3. Yol, A. M.; Dabney, D. E.; Wang, S-F.; Laurent, B. A.; Foster, M. D.; Quirk, R. P.; Grayson, S. M.; Wesdemiotis, C. “Differentiation of Linear and Cyclic Polymer Architectures by MALDI Tandem Mass Spectrometry (MALDI-MS2)” J. Am. Soc. Mass Spectrom. 2013, 24, 74-82. 4. Quirk, R.P.; Chavan, V.; Janoski, J.; Yol, A.; Wesdemiotis, C. “General Functionalization Method for Synthesis of α-Functionalized Polymers by Combination of Anionic Polymerization and Hydrosilation Chemistry” Macromolecular Symposia 2013, 323, 51-57. 5. Yol, A. M.; Janoski, J.; Quirk, R. P.; Wesdemiotis, C. “Sequence Analysis of Styrenic Copolymers by Tandem Mass Spectrometry” Anal. Chem. (Submitted) v DEDICATION To the memory of my father and mother. To all my brothers, sisters, nephews, nieces, brothers-in-law, and sisters-in-law. vi TABLE OF CONTENTS Page LIST OF TABLES………………………………………………………...........................x LIST OF FIGURES………………………………………………………………………xi LIST OF SCHEMES……………………………………………………………………xvi CHAPTER I. INTRODUCTION………………………………………………………………...1 II. MASS SPECTROMETRY BACKGROUND…………………………………….5 2.1. Ionization techniques…………………………………………………………5 2.2. MALDI……………………………………………………………………….5 2.3. Mass analyzers………………………………………………………………..8 2.3.1 Mass resolution……………………………………………………..9 2.3.2 Time of fight (ToF) mass analyzer………………………………...10 2.3.2.1. Reflectron………………………………………………..12 2.3.2.2. Delayed extraction (DE) or pulsed ion extraction (PIE) or time lag focusing……………………………………………........13 2.4. Detectors…………………………………………………………………….15 2.4.1. Microchannel plate (MCP) detector……………………………….15 III. MATERIALS AND EXPERIMENTAL PROCEDURES………………………17 3.1. Materials…………………………………………………………………….17 3.1.1. Linear and cyclic polymers…..........................................................17 3.1.2. Polystyrene copolymers………………...…………………………20 vii 3.1.3. Linear in-chain functionalized precursor, cyclic with two linker units, and four star polystyrenes………………........................................20 3.2. MALDI experimental procedures………………………………………...…21 3.3. MALDI instrumentation….............................................................................22 IV. DIFFERENTIATION OF LINEAR AND CYCLIC POLYMER ARCHITECTURES BY MALDI TANDEM MASS SPECTROMETRY (MALDI-MS2)…………………………………………………………………...24 4.1. Linear Polystyrene…………………………………………………………..24 4.2. Cyclic Polystyrenes…………………………………………………………30 4.3. Cyclic Polybutadiene……………………………………………………......43 4.4. Conclusions…………………………………………………………….…....48 V. SEQUENCE ANALYSIS OF STYRENIC COPOLYMERS BY TANDEM MASS SPECTROMETRY………………………………………………………49 5.1. Composition and architecture of poly(dimethylsilylstyrene-co-styrene) copolymers ………………………………………………………………………49 5.2. Reference MS2 spectra of polystyrene and poly(p-DMSS-b-styrene)………54 5.3. Sequence analysis of poly(p-DMSS-co-styrene) and poly(m-DMSS-co- styrene)…………………………………………………………………………...61 5.4. Conclusions….................................................................................................68 VI. MALDI-TOF/TOF TANDEM MASS SPECTROMETRY OF LINEAR IN- CHAIN SUBSTITUTED, CYCLIC WITH TWO LINKER UNITS, AND FOUR- ARM STAR-BRANCHED POLYSTYRENES……….………………………...69 6.1. Linear in-chain substituted PS…………………………………………...….69 6.2. Cyclic PS with two linker units……………………………………………..74 6.3. 4-arm star-branched polystyrene…………………………………………….79 6.4. Conclusions………………………………………………………………….87 VII. SUMMARY……………………………………………………………………...88 viii REFERENCES…………………………………………………………………..90 APPENDIX………………………………………………………………………98 ix LIST OF TABLES Table Page 2.1. Common MALDI matrices…………………………………………………………..7 2.2. Common lasers used for MALDI experiment………………………………………..8 5.1. Measured vs. calculated monoisotopic m/z values of the oligomers observed in the low (m/z 1490-1630) and high (m/z 3530-3710) mass regions (Figures 5.2b-c) of the MALDI mass spectrum of poly(p-dimethylsilylstyrene-co-styrene).……………………53 x LIST OF FIGURES Figure Page 2.1. MALDI principle…………………………………………………………………......6 2.2. Resolving power…………………………………………………………………….10 2.3. Linear time of fight principle………………………………………………………..11 2.4. Principle of reflectron ToF instruments (reproduced with permission from ref. 76). The filled circle represents the faster moving ion ……………………………………….13 2.5. Principle of continuous and delayed extraction……………………………………..14 2.6. Microchannel plate (MCP) detector…………………………………………………16 3.1. Bruker utraFlex III mass spectrometer (reproduced with permission from ref. 84). IS1 and IS2 are ion source lenses; PCIS is the precursor ion selector (also called timed ion + + selector, TIS); P1 and P2 are two ion families, each composed of a precursor ion and its fragments