1. Methylation of acidic moieties to enable characterization in MAA based copolymer Rémi Giordanengo, (1) Stéphane Viel, (1) Manuel Hidalgo, (2) Béatrice Allard-Breton, (2) André Thévand (1) and Laurence Charles (1) (1) Aix-Marseille University – Marseille, France (2) ARKEMA – Centre de Recherche Rhônes-Alpes – Pierre-Bénite, France 58 th ASMS Conference, May 24, 2010, Salt Lake City

2. Copolymer Characterization by Mass Spectrometry 2 Molecular Weight M n, M w Nature of end-groups X,Y Composition n, m Nature of MAA-MMA copolymer block vs random MAA M = 86 Da MMA M = 100 Da Two ionization techniques : ESI and MALDI

3. 499 513 527 599 613627 699 713 727 ESI-MS 3 ESI(+)-MS spectrum 2 distributions : [M + Na] + [M + 16 + Na] + ∆ m/z 100 ∆ m/z 86 ∆ m/z 100 ∆ m/z 86 MMA MAA

4. ESI-MS 4 ESI(+)-MS spectrum 499 513 527 599 613 627 699 713 727685 413 427 327 313 M X, M Y unknown n, m ?

5. ESI-MS/MS MS/MS requested to determine the actual values of n and m Dissociation rules of MAA-MMA copolymers in negative and positive mode ESI : water elimination methanol elimination Between two consecutive neutral MAA → MAA/MAA pair in the polymeric chain Between neutral MAA and MMA→ MAA/MMA pair in the polymeric chain The number of water and methanol losses is monitored to reveal the number and arrangement of MAA and MMA units in the selected precursor ion Giordanengo, R. et al. J. Am. Soc. Mass Spectrom. 2009, 20 (1), 25-33 ; Giordanengo, R. et al. Rapid Commun. Mass Spectrom. 2009, 23 (11) 1552-1567 ; Giordanengo, R. et al. Anal. Chim. Acta 2009, 654, 49-58 Na + 5

6. Only one loss of methanol Only one MAA/MMA pair 1 MMA unit m+1 = 1 ESI-MS/MS 6 Neither water nor methanol loss in MS/MS spectra of precursor ion in the first column No MAA unit n = 0 - H 2 O - MeOH Only one loss of water Only one MAA/MAA pair 2 MAA units m/z 499 (2 ; m+1)

7. ESI-MS/MS 7 M X + M Y = 204 Da m/z 689 m/z 771 m/z 753 m/z 739 m/z 735 m/z 721 m/z 707 - H 2 O - MeOH 2 consecutive losses of H 2 O 2 MAA-MAA pairs 4 MAA units 1 loss of H 2 O 2 losses of MeOH 1 MAA-MAA pair 2 MAA-MMA pairs 2 MMA units Different dissociation routes from the same precursor ion Random nature of the studied MAA-MMA copolymer m/z 771 (4;2) M X and M Y unknown

8. Limitations of the ESI-MS/MS Approach 8 Dissociation of MAA-MMA co-oligomer sodiated adducts allows : To reach the co-monomeric composition of co-oligomers To reveal the random nature of the studied MAA-MMA copolymer To determine the sum of the end-group masses (M X + M Y ) However : The lack of specific fragmentation does not allow the end-groups to be structurally characterized (contrary to what is usually obtained in the case of homopolymers such as PMMA) MAA-MMA ESI-MS(/MS) (n, m) Nature (M X + M Y )

9. Same phenomenon in ESI of constitutive homopolymers PMMA and PMAA PMMA 1590PMAA 1250 Signal only in the low m/z range indicating a strong bias toward low mass oligomers Other ESI-MS Limitations 9 ESI-MS spectrum of the MAA-MMA copolymer : Distribution parameters largely underestimated as compared to NMR data (M n 3100 Da) Strong bias toward low mass oligomers for all acrylic polymers Wrong values of M n and M w Uncorrect MAA/MMA ratio M w (1+) 865 M w (2+) 1592 M w (1+) 612 M w (2+) 728 Haddleton, D. M.; Feeney, E.; Buzy, A.; Jasieczek, C. B.; Jennings, K. R., Chemical Communications 1996, (10), 1157-1158.

10. PMMA 1590PMAA 1250 10 MALDI-MS : In contrast, MALDI-MS spectra of constitutive homopolymers PMMA and PMAA are more realistic However, no interpretable signal obtained in MALDI for the MAA-MMA copolymers Limitations in Terms of Molecular Weight Determination

11. Strategy 11 In contrast to results obtained for MAA-MMA copolymers: End-groups characterization is possible during CID of PMMA homopolymers Distribution parameters can be achieved by MALDI of PMMA homopolymers STRATEGY Transform MAA-MMA copolymers into PMMA homopolymers by methylating all MAA units ~ 50 mg of MAA-MMA copolymer in 20 mL of a MeOH/H 2 O mixture (19/1, v/v) (stirred 2h at room temperature) Addition of an excess of trimethylsilyldiazomethane (stirred 3h at room temperature) Drying at 30°C overnight Experimental Couvreur, L.; Lefay, C.; Belleney, J.; Charleux, B.; Guerret, O.; Magnet, S. Macromolecules 2003, 36, 8260-826

12. Strategy 12 Derivatization of MAA-MMA copolymers to determine distribution parameters

13. Methylation of PMAA Homopolymers 13 MALDI(+)-MS spectrum PMAA 1700 methylated PMAA 1700 Methylation - Complex MS spectrum due to the ability of PMAA to bind numerous Na + cations. - High laser fluence (65%). - A single PMMA distribution is obtained, indicating all acrylic acid functions of PMAA have been methylated. - Good signal-to-noise ratio is measured from low laser fluence (~ 40%). PMAAMethylated PMAA PMAAM w SEC M w MALDI M w SEC M w MALDI M w PGSE 112501132144512311452 217001487196616341701 331501951365425792928 466004546766456206311

14. 14 200035005000 400 800 No signal obtained before methylation 2000 4000 6000 600 200 1000 After methylation Intense MALDI-MS signal M w can be determined by MALDI and PGSE NMR Giordanengo, R.; Viel, S.; Hidalgo, M.; Allard-Breton, B.; Thevand, A.; Charles, L. J. Am. Soc. Mass Spectrom. 2010, 21 (6) 1075-1085. Methylation for Molecular Weight Determination Methylated CopolymerCopolymer M n MALDI M n NMR M w MALDI M w PGSE M w calculated from M w MALDI M w calculated from M w PGSE MAA/MMA 3200315043605170 41104870

15. Strategy 15 Derivatization of MAA-MMA copolymers for end-group characterization

16. Methylation for End-Group Characterization 16 [M + Na] + (M X + M Y ) = 204 [M + 16 + Na] + (M X + M Y ) = 220 [M + Na] + (M X + M Y ) = 204 [M + 16 + Na] + (M X + M Y ) = 220 Methylation MAA-MMA methylated MAA-MMA After methylation : ESI-MS simplified A single PMMA distribution is obtained indicating all MAA units have been methylated No modification of the end-groups during the methylation reaction ESI(+)-MS :

17. 17 ESI(+)-MS/MS of the methylated MAA-MMA copolymer : m/z 827 Dissociation rules of PMMA homopolymers in positive mode ESI (1) : Series FSeries A Series B Series D C+C+ C+C+ C+C+ C+C+ C+C+ C+C+ C+C+ Methylation for End-Group Characterization Jackson, A. T.; Yates, H. T.; Scrivens, J. H.; Critchley, G.; Brown, J.; Green, M. R.; Bateman, R. H. Rapid Commun. Mass Spectrom. 1996, 10, 1668 ; Jackson, A. T.; Yates, H. T.; Scrivens, J. H.; Green, M. R.; Bateman, R. H. J. Am. Soc. Mass Spectrom. 1997, 8, 1206

18. 18 ESI(+)-MS/MS of the methylated MAA-MMA copolymer : Dissociation rules of PMMA homopolymers in positive mode ESI (2) : C + C+C+ C+C+ Series DSeries C C+C+ C+C+ C+C+ Series FSeries E Methylation for End-Group Characterization Jackson, A. T.; Yates, H. T.; Scrivens, J. H.; Critchley, G.; Brown, J.; Green, M. R.; Bateman, R. H. Rapid Commun. Mass Spectrom. 1996, 10, 1668 ; Jackson, A. T.; Yates, H. T.; Scrivens, J. H.; Green, M. R.; Bateman, R. H. J. Am. Soc. Mass Spectrom. 1997, 8, 1206

19. 19 Application of the dissociation rules of PMMA homopolymers in positive mode ESI : m/z 827 Most intense fragment ions: Series A or B The two most intense fragment ions could be identified to Series A, revealing Y = H Na + A 0 : m/z 110 A 1 : m/z 210 A0A0 A1A1 Methylation for End-Group Characterization

20. Na + E 0 : m/z 209 E 1 : m/z 309 E 2 : m/z 409 E 3 : m/z 509 E 4 : m/z 609 Series E Validation of the nature of Y M Y = 1 Y = H 20 Application of the dissociation rules of PMMA homopolymers in positive mode ESI : m/z 827 Since Y = H, product ions D and E can be assigned Na + D 2 : m/z 265 D 3 : m/z 365 D 4 : m/z 465 D 5 : m/z 565 Series D Methylation for End-Group Characterization A0A0 A1A1

21. Na + Series F F 1 : m/z 381 F 2 : m/z 481 F 3 : m/z 581 Na + Series B B 0 : m/z 226 B 1 : m/z 326 Na + Series C C 1 : m/z 425 C 2 : m/z 525 C 3 : m/z 625 C 4 : m/z 725 21 Application of the dissociation rules of PMMA homopolymers in positive mode ESI : M X + M Y = 204, with M Y = 1 → M X = 203 Based on M X = 203, product ions B, C and F can be identified Methylation for End-Group Characterization Validation of M X = 203

22. Na + Use of a sulfurated agent during the copolymer synthesis, 2-ethylhexyl thioglycolate (M = 204 Da) 2 additionnal product ions detected at m/z 715 et m/z 683 allow to validate the structure proposed for X 22 Proposed structure for X 715 683 Methylation for End-Group Characterization Proposed structure for X

23. X end-group Nature δ 13 C (ppm)δ 1 H (ppm) 1 2 3 4 1’ 2’ 3’ 4’ 5 CH 3 CH 2 CH 3 CH 2 CH CH 2 C(O) 14.6 24.0 30.0 31.4 11.4 24.7 40.0 68.3 172.0 0.96 1.38 1.37 1.40 0.98 1.45 1.65 4.10 - 6767 CH 2 35.5 45.9 3.27 2.93 2.75 NMR Analysis 23 Validation of the structure of the end-groups Proposed structure of X : NMR 1D ( 1 H, 13 C) and 2D (COSY, HSQC, HMBC, DOSY) analysis Giordanengo, R.; Viel, S.; Hidalgo, M.; Allard-Breton, B.; Thevand, A.; Charles, L. Rapid Commun. Mass Spectrom., in press

24. Conclusion 24 Methylation of MAA-MMA copolymers allows : A complete microstructural characterization The distribution parameters to be determined (with the help of NMR data) consistently with NMR results MAA-MMA ESI-MS(/MS) (n, m) Nature (M X + M Y ) Methylation M X, M Y MAA-MMA MALDI-MS failed Methylation M n, M w

25. Acknowledgments 25 The organisation committee The Spectropôle, analytical facility of Aix-Marseille University, for the special access to the instruments ARKEMA, for the financial support SACS and C&S members Thank you for your attention