APC Hyphenated to YAPT-G2: Potential For The Analysis f Complex ynthetic Polymers Marie-Theres Picker Artjom Döring Dirk Kuckling International ymposium on GPC/EC and Related Techniques Amsterdam, 2016
University of Paderborn: founded in 1972 currently more than 20.000 students material science profile in chemistry, physics and mechanical engineering Paderborn 2
Research Interests of Prof. Kucklings Working Group: X Controlled Radical Polymerization (MRP, ATRP, RAFT) mart block copolymers for micro heterogeneous organocatalysis or pigment stabilization Y + Z mart Polymers MR IR & UV/VI PR Analytics EC, APC M (MALDI, EI-IM) Free Radical Polymerization and photo cross-linking / photo patterning Hydrogels for applications in biomedicine and as actuators and sensors in micro-system technology mart and biodegradable block copolymers for triggered release systems mart nanohydrogels for targeted transport and controlled release systems Multisensitive phase separated hydrogel layers Receptor/Ligand modified (hydro)gels for sensor applications enzyme 3
Applied eparation Techniques ize Exclusion Chromatography (EC) - fractionation of the polymer sample according to the hydrodynamic volume - molecular mass distribution as a result after calibration (relative, universal, absolute) Advanced Polymer Chromatography (APC) - UHP-EC - significantly reduced measurement times and sample concentration - robust porous silica matrix Electron pray Ionization Mass pectrometry (EI-M) - soft vaporization and ionization of macromolecules - separation according to mass/charge ratio - less complicated sample preparation - determination of repeating units and end groups in macromolecules - fairly complex M spectra (mass dispersity, chemical heterogeneity, charge dispersity, mixed ionization) Ion Mobility eparation (IM) - separation of ions with respect to charge, shape and size APC-EI-IM Hyphenation Pacholarz et al., Chem. oc. Rev., 2012, 41, 4335 4
Experimental etup Quadrupole analyzer Triwave collision cell QuanTF analyzer Waters APC Columns: Acquity APC XT 125Å & Acquity APC XT 45Å Eluent: THF/DMF (v/v 80:20) ai (68 µm) Flow rate: 0.7 ml/min Injection volume: 20 µl AI Quickplit Adjustable Flow plitter (ratio approx. 95:5) EI source MALDI source 5
Performance of the Waters APC -ystem Eluent: THF/DMF (v/v 80:20), ai (68 µm) DMF as polar solvent for EI ai as ionizing agent no need of secondary pump for addition of make-up solvent and ionizing agent Comparison of APC and EC results revealed ize Exclusion for: PMMA -substituted polyacrylamides (PIPAAm, PDMAAm) Polycarbonates Advanced Polymer Chromatography Columns: Acquity APC XT 125Å & Acquity APC XT 45Å Eluent: THF/DMF (v/v 80:20), ai (68 µm) Flow rate: 0.7 ml/min Injection volume: 20 µl Traditional ize Exclusion Chromatography Columns: P DV (10 6 Å, 10 5 Å, 10 3 Å, 10 2 Å) Eluent: CHCl 3 (0.1 Vol% TEA) Flow rate: 0.75 ml/min Injection volume: 50 µl 6
Challenges for the Waters APC -ystem CH m n o co Boc H H source: www.waters.com source: www.pss-polymers.com need of modified silica matrices for APC columns polarity fine-tuning is necessary to ensure pure size exclusion without enthalpic interactions 7
Complexity of polymer samples Macromolecules exhibit different types of heterogeneity: Distribution of molar mass (monomodal, bimodal etc.) Chemical Composition (copolymers: statistic, gradient, block) Topology (linear, (hyper)branched, dendritic, star, graft, cross-linked) Functionality (end groups, side groups, cyclic) Additionally, mixtures of different polymers can significantly increase sample complexity. Analyzed samples (prepared by RAFT-polymerization) poly(,-dimethylacrylamide)-bpoly(-isopropylacrylamide) with residual amounts of poly(,dimethylacrylamide) homopolymer Mixture of a linear and 4-arm-star homopolymer based on poly(,-dimethylacrylamide) 8
Analysis of poly(,-dimethylacrylamide)-b-poly(-isopropylacrylamide) HC n EG EG = HC m n H H M n,apc = 6900 g/mol D = 1.38 AIB 1.4-dioxane, 70 C EG = unreactive (olefine, saturated) M n,apc = 3700 g/mol D = 1.15 HC n EG poly(,-dimethylacrylamide)-bpoly(-isopropylacrylamide) with residual amounts of poly(,dimethylacrylamide) homopolymer 9
Analysis of poly(,-dimethylacrylamide)-b-poly(-isopropylacrylamide) with APC-hyphenation without APC-hyphenation EI source Capillary: 3.0 kv ampling cone: 120 V Extraction cone: 3.0 V IM cell Wave length: 550 m/s Wave height: 40 V 10
Analysis of poly(,-dimethylacrylamide)-b-poly(-isopropylacrylamide) with APC-hyphenation without APC-hyphenation
species 4 with APC-hyphenation species 2 species 1 species 3 without APC-hyphenation number of species? species 4 species 2 species 3 species 1 12
species 4 with APC-hyphenation species 2 species 1 species 3 Δm/z = 0.05 Δm/z = 0.05 HC 16 9 H a+ K+ HC 23 3 H a+ a+ 13
with APC-hyphenation HC species 4 species 2 species 1 species 3 16 9 H a+ K+ M avg, calc = 2969.156 g/mol M avg, found = 2968.833 g/mol C 15 10 H a+ K+ M avg, calc = 2964.182 g/mol M avg, found = 2964.221 g/mol HC 23 16 H a+ a+ K+ M avg, calc = 4455.180 g/mol M avg, found = 4456.315 g/mol C 14 24 H a+ K+ K+ M avg, calc = 4449.262 g/mol M avg, found = 4449.567 g/mol 14
Analysis of a mixture of a 4-arm star and a linear homopolymer of poly(,-dimethylacrylamide) RAFT polzmeriyation with a 4-arm trithiocarbonate chain transfer agent PD = 1.13 M n = 4,000 g/mol 1.4-dioxane 70 C M n = 4,400 g/mol PD = 1.06 M Addition of linear n = 4,500 g/mol PD = 1.04 poly(,-dimethylacrylamide) homopolymer m n p + H n o 15
Analysis of a mixture of a 4-arm star and a linear homopolymer of poly(,-dimethylacrylamide) with APC-hyphenation without APC-hyphenation EI source Capillary: 3.0 kv ampling cone: 120 V Extraction cone: 3.0 V IM cell Wave length: 550 m/s Wave height: 40 V 16
with APC-hyphenation without APC-hyphenation 17
Analysis of a mixture of a 4-arm star and a linear homopolymer of poly(,-dimethylacrylamide) with APC-hyphenation contrary tendency compared to linear and star-shaped PEG polymers (Foley et al., AC Macro Lett. 2015, 4, 778) series of triply charged species + + + more expanded structure of the star polymer due to minimizing the electrostatic repulsion 18
ummary APC respresents a robust system to determine MWDs within significantly shorter measurement times and without loss of separation efficiency APC-hyphenation to ynapt G2 offers fractional sample injection into the M-ystem temporal resolution of M spectra significant improve of signal/noise ratio significantly better resolution for the drift time dimension in IM APC-hyphenation helps to ionize structurally different macromolecules in a mixture APC-hyphenation complements EI-IM-M separation, thus giving the opportunity to have more detailed insight into complex polymer samples 19