Mass Spectrometry DOI: 10.1002/anie.200((will be filled in by the editorial staff)) Generation of Gas-Phase Sodiated Arenes [(Na3(C6H4)+], and Other Unusual Sodium Adducts Such As (Na3O)+, and (Na2OH)+ Athula B. Attygalle,* Chang-Ching Chan, Frank U. Axe and Mark S. Bolgar Figure 2A is much higher than those in Figure 2B and 2C, which indicated that the sodium ion is less tightly bound to the rest of the molecule in the ortho isomer even after the first CO2 is lost (See also Figure S2). 63 Na 100 + + H O Na Na Na O O 189 O % A CO2 Na+ 23 Na O 233 0 + Na 63 Na 100 + H O O % B 0 Na + Na O Na+ 23 O CO2 Na Na O 189 145 CO2 166 167 Na 85 233 + 100 O CO2 % 0 85 50 75 CO2 145 166 167 Na 25 Na 189 Na + Na O Na+ 23 Na O + H Na C Na O O 63 Na 100 125 150 175 O O 233 200 225 Na m/z m/z 250 Figure 1. Product-ion spectra of m/z 233, the sodium adduct of disodium phthalate (A), isophthalate (B), and terephthalate (C) recorded on a Quattro-Micro mass spectrometer (collision energy = 20 eV). Moreover, the m/z 189 ion from the meta and para isomers loses another molecule of CO2 to produce the ion observed at m/z 145 which still retains three sodium atoms. In contrast, the m/z 189 from the ortho isomer failed to show a further CO2 loss (Figure S3). Na+ 23 100 A + CO2 Na Na. Na + H O % Collision-induced dissociation (CID) mass spectra of metal cationized peptides, nucleic acids and lipids[1-3] are known to provide structural information complementary to those recorded from their protonated counterparts.[4-8] However, very few investigations have been undertaken on fragmentation of cationic adducts of acidic small molecules. Generally, CID investigations of cationized small molecules are less rewarding since most spectra show only a single product ion peak for the ejected metal cation. For example, the ESI spectrum of sodium benzoate shows peaks at m/z 167 and 311 for the ion/neutral complexes formed between the neutral disodium salt and sodium ion. Such adducts, when subjected to CID, readily ejects the neutral salt to produce a peak at m/z 23 for the sodium cation (Figure S1, Supporting Information). However, when the metal atom binds in a multi-dentate fashion to several heteroatoms simultaneously, as seen with many carbohydrates, the product ions are known to retain the metal atom.[9-11] During our explorations on fragmentation mechanisms of low-molecular-weight carboxylic acids,[12] we noted that the alkali adducts of the disodium salts of dicarboxylic acids upon CID undergo specific fragmentation reactions in which the sodium atom is often retained. Under CID conditions, sodium adducts of disodium salts of all isomers of benzene dicarboxylic acids fragmented in an unprecedented manner (see Supporting Information for experimental details). Instead of expelling a Na+, all of them ejected CO2 to produce an m/z 189 ion that retained all three sodium atoms of the precursor (Figure 1). Although not directly related, an ejection of CO2 from the phenyl anion derived from sodium terephthalate had been previously observed only under negative ion conditions.[13] Moreover, the product ion spectrum recorded from the disodium phthalate adduct contained fewer peaks compared to those from its two positional isomers indicating the presence of a significant ortho effect (Figure 1). Tandem mass spectrometric experiments conducted with the m/z 189 ion produced by in-source fragmentation demonstrated that the charge-imparting sodium ion is more securely bound to the meta and para isomers than to the ortho isomer. For example, the relative abundance of the m/z 23 peak in Na Na 63 122 166 O O Na 189 0 [] O Na O Na. Na O 166 Na Na+ 23 0 Na 145 Na B + CO2 Na. + H Na + Na O % Prof. A. B. Attygalle Department of Chemistry and Chemical Biology, Stevens Institute of Technology, Hoboken, NJ 07030 (USA) Fax: (+001) 201-216-8240 E-mail: aattygal@stevens.edu Homepage: http://www.stevens.edu/ses/chembio/MassSpectrometry/ Chang-Ching Chan, Dr. Mark S. Bolgar Bristol-Myers Squibb Analytical Research & Development, 1Squibb Drive, New Brunswick, NJ 08903 (USA) Dr. Frank U. Axe Axe Consulting Services, 14595 Surrey Junction Lane, Sutter Creek, CA 95685 (USA) This research was supported by Bristol-Myers Squibb Analytical Research and Development and Stevens Institute of Technology, NJ 122 85 189 CO2 63 100 Na + H O C 20 40 60 80 100 120 140 Na Na O 166 122 85 O Na. Na Na+ 23 0 + Na Na. 145 Na Na + Na O % [] 63 Na 100 160 189 180 m/z Figure 2. Product ion spectrum of m/z 189 ion formed by in-source fragmentation of the sodium adduct of disodium phthalate (A), isophthalate (B), and terephthalate (C) recorded on a Quattro-Micro mass spectrometer (collision energy = 25 eV). Supporting information for this article is available on the WWW under http://www.angewandte.org or from the author. 1 In the optimized structure predicted by DFT calculations for the m/z 233 ion from the ortho isomer (1), two of the sodium atoms interact in a bidentate fashion with each carboxylate group, while the third sodium atom bridges the two carboxylate groups (Scheme 1). sodium atom bridges the two carboxylate groups. The bridging interaction appears to distort the aromatic ring carbons attached to the carboxylate group slightly out of plane particularly for the meta and para isomers by 10° and 20°, respectively. In addition, the bridging sodium atom particularly in the para isomer interacts with the electrons of the benzene ring (6) (Scheme 3). In contrast to that from the ortho isomer, the m/z 189 ions derived from both meta and para-isomers undergo a second CO2 loss, to produce a peak at m/z 145 indicating that the CO2 residue in the m/z 189 ion derived from both meta and para-isomers is not as securely bound as that of the ortho isomer’s. The optimized structures computed for the m/z 189 ions from meta and para isomers are shown on Schemes 2 and 3. CO2 Scheme 1. Fragmentation of sodium adduct of disodium phthalate m/z 189 [Na3C6H4(CO2)+] (7) Once one of the carboxylate groups is eliminated, the sodium atoms in the residual molecule are predicted to interact with the main structural skeleton in three distinct ways (2). First, one of the sodium atoms interacts in a bidentate fashion with the remaining carboxylate group similar to that seen in the trisodium precursor. Second, one of the sodium atoms is involved in a two-center twoelectron bond with the phenyl carbon, from which the carboxylate group was eliminated, which also interacts with the remaining carboxylate group as well. Third, the remaining sodium atom in the ortho isomer interacts with all six carbon atoms of the phenyl ring in a 6 fashion.[14] Since one of the oxygen atoms in the remaining carboxylate group is also coordinated to the sodium atom bound to the adjacent carbon atom of the benzene ring, the structure receives considerable enhanced stability. Consequently, it is resistant to further CO2 loss. In other words, upon CID the sodium ion that is loosely coordinated by 6 cation-π interactions to the benzene ring gets ejected with minimal collisional activation, rather than losing a second CO2 molecule. The coordination of sodium atoms in the precursor meta and para isomers (3 and 6, respectively; Schemes 2 and 3) is somewhat similar to that observed for the ortho isomer (1). Scheme 2. Fragmentation of sodium adduct of disodium isophthalate For example, two of the sodium atoms in the sodium adducts of disodium isophthalate (3) and terephthalate (6) interact in a bidentate fashion with each carboxylate group, while the third m/z 233 [Na3C6H4(CO2)2+] (6) CO2 m/z 145 [Na3(C6H4)+] (8) Scheme 3. Fragmentation of sodium adduct of disodium terephthalate In both structures 4 and 7, one sodium atom is bonded by a twocenter-two-electron bond to one of the phenyl ring carbons (in contrast to that observed for the ortho isomer, this is an isolated bond), one sodium atom interacts in a bidentate fashion with the remaining carboxylate group, while the charge-bearing sodium atom is simultaneously coordinated to electrons of the benzene ring and one of the oxygen atoms of the remaining carboxylate group. In other words, the charge-bearing sodium atom in structures 4 and 7 are more tightly coordinated to the main skeleton than that in structure 2. Consequently, m/z 189 ion derived from the ortho isomer (2) does not undergo a second CO2 loss, in contrast to those from meta and para isomers which eliminate a second CO2 to produce ions 5, and 8, respectively. The molecular formula of the m/z 145 ion produced when a second CO2 is eliminated from meta and para isomers is Na3C6H4+, which in fact represents the sodium adduct of phenelenedisodium, an ionic species which has not been reported previously. Although, aryl derivatives of the alkali metals are known as useful strong bases and potent nucleophiles,[15] only on one rare occasion has phenelenedisodium been mentioned in the literature.[16] On the other hand, 1,4- and 1,3-dimetalations have been accomplished with “spiny” arenes such as tert-butylbenzene.[17] The observation of Baston et al[17] that 1,2-isomers of dimetalloarenes are not formed at all, is congruent with our result of not detecting a peak for Na3C6H4+ from ortho compounds. Moreover, the m/z 145 sodium adducts we report here correspond to the lithium and sodium salts of double-deprotonated benzene that have been observed by Kass and his co-workers under negative ion conditions.[13,18] Moreover, ortho ions have been observed to behave different under negative ion conditions.[19] To comprehend the unique fragmentations reported here, it is necessary to understand the mechanism that allows retention of the sodium atom originally coordinated to the carboxylate group when a CO2 molecule is eliminated. For this elimination to occur, the 2 coordinating sodium ion must pirouette around the carboxylate group and insert into the Caro-CO2 bond (Scheme 4). This process was analyzed with DFT calculations for the sodium adduct of the disodium salt of 1,4-benzene dicarboxylic acids (m/z 233) and found to be reasonable, although all processes are energetically “up-hill” (Figure S4). On the other hand, in the m/z 189 ion from the ortho compound (2), the inserted sodium atom ends up coordinating both the aromatic carbon and the remaining carboxylate oxygen which renders the second CO2 elimination much more difficult by this mechanism. mechanism for the ortho isomer is depicted in Scheme 6. In this mechanism the bidentate coordinated sodium ion can swing around the carboxylate group (red arrow) and insert into the C-CO2 bond while displacing the bridging sodium ion into a -coordinating position on the aromatic ring (blue arrow). Considering the observation that a second CO2 loss was disfavored without the loss of sodium radical, it is plausible that eliminating the 6 sodium ion provides room for the remaining two sodium ions (2a) to rearrange and allow the second CO2 elimination to occur. + + Na O O Na Na -CO2 O O Na O O O Scheme 4. Fragmentation of sodium adduct of disodium terephthalate (m/z 233). Another intriguing observation that was made upon collision activation was that all the three isomers of the m/z 189 ion expelled a sodium radical to produce a peak at m/z 166 for a distonic radical cation [(●C6H4CO2Na2)+] (Figure 2). However, the meta and para isomers expelled the sodium radical to produce the m/z 166 radical cation (10 and 11) more readily than the ortho isomer. Apparently, the m/z 189 ion from the ortho isomers prefers to undergo a heterolytic fragmentation to produce a sodium ion by eliminating a neutral (12). + Na Na O CO2 Na C Na2C6H4+ O + Na O C m/z 122 Na m/z 166 (ortho, 9) (meta, 10) (para, 11) O Na O m/z 189 (ortho, 2) (meta, 4) (para, 7) C + Na m/z 23 Na O Na 166 Da (12) Scheme 5. Fragmentation of m/z 189 ion from ortho, meta and para isomers. In order to rationalize these differences, we calculated the energetics of neutral sodium atom and sodium cation elimination from isomeric m/z 189 ions (Scheme 5). The relative energies obtained for these two competing processes demonstrate that the elimination of the sodium atom directly bonded to phenyl ring as a neutral atom is preferred for both meta and para isomers (4 and 7), while for the ortho isomer elimination of the 6-bonded sodium atom as a cation is favoured (Table 1). m/z 189 (2) m/z 166 (2a) The m/z 166 radical cation [(●C6H4CO2Na2)+] for all three isomers then eliminates CO2 to produce a peak at m/z 122 corresponding to the distonic cation (●C6H4Na2)+. The predicted m/z 122 (2b) Scheme 6. Fragmentation of m/z 189 derived from the sodium adduct of disodium phthalate (m/z 233). Also, our DFT calculations on both the m/z 166 and 122 ions clearly predict that the unpaired electron resides primarily on the coordinatively unsaturated aromatic carbon in all cases. Two other unprecedented peaks in the spectra of sodium adducts of disodium benzene dicarboxylates are those observed at m/z 63 and 85 (Figure 1 and 3). The peak at m/z 63, which appeared to represent sodiated sodium hydroxide, was particularly intriguing since a proton is required for the formation of this charged species. An abstraction of a proton from the aromatic nucleus of the benzene dicarboxylate system is an energetically unfavorable process since deprotonation energy of benzene to be about 402 kcal/mol. [20] Moreover, the product ion spectrum of sodium adduct of disodium [2,3,5,6-2H4]terephthalate showed no sign for a peak at m/z 64 for [Na2OD]+, which confirmed that a proton is not extracted from the aromatic system for the formation of this ion (Figure 3). In fact, the proton originates from an unprecedented ion-molecule reaction that has taken place with residual water in the collision cell. This was confirmed by the observation of a peak exclusively at m/z 64 for [Na2OD]+ (Figure S5) when product-ion spectra of m/z 233 ion were recorded with D2O vapor introduced to the collision gas of the QToF tandem mass spectrometer. Since the relative abundance of the m/z 63 peak was significantly higher in the tandem mass spectra recorded from the m/z 189 ion produced by in-source fragmentation of the meta- and para-isomers than that from the ortho isomers, it appears that the m/z 189 ion structures proposed from the meta and para-isomers show a higher affinity to water than that projected for the ortho isomer. 149 100 + O 193 63 % D Na Na. D D CO2 O Na. Na+ 23 Na + Na O Na 85 0 Na O D Na + H O Na Table 1: Relative elimination energies calculated for ortho, meta, and para isomers pertaining to the reaction, Na2C6H4(CO2)+ + Na• → Na2C6H4(CO2)• + Na+. Isomer E (kcal/mol) ortho -4.87 meta +4.77 para +3.81 Na Na Na O Na Na Na O O Na Na Na CO2 + Na 25 50 75 100 CO2 O Na 170 126 171 237 125 150 175 200 225 m/z 250 Figure 3. Product-ion spectrum of m/z 237 for the sodium adduct of disodium [2,3,5,6-2H4]terephthalate recorded on a Quattro-Micro mass spectrometer at a collision energy setting of 20 eV. 3 We propose that the initial ion/water complex (13) rapidly rearranges into a transitional complex between NaOH and the sodium adduct of sodium benzoate (15) (Scheme 7), which either undergoes a direct dissociation to yield the ion observed at m/z 167 (which becomes m/z 168 if D2O is present in the collision gas), or endure a transfer of the Na+ to NaOH to form the transitional complex 16. Nicholas et al. have calculated that the energy barrier between systems in which a sodium cation binds to either an oxygen or a phenyl ring are very similar; hence the Na+ could be expected to be mobile from one binding site to the another.[21] A subsequent, separation of the constituents in the complex 16 leads to [Na2OH]+ (17) and sodium benzoate, or [Na3O]+ (18) and benzoic acid (Scheme 7). It is interesting to note that compounds such as Na3O(NO2) and Na3OBr have been reported previously.[22] H O H Na+ H Na O H H H Na+ Keywords: radical ions · gas–phase chemistry · laser ablation · silver · electron capture [1] A. Favre, F. Gonnet, J.-C. Tabet, Int. J. Mass Spectrom. 1999, 190/191, 303. [2] Y. Wang, J-S. Taylor, M. L. Gross, J. Am. Soc. Mass Spectrom. 2001, O-Na+ Na O Spectrom. 2003, 38, 87. + Na [4] N. C. Polfer, J. Oomens, R. C. Dunbar, ChemPhysChem 2008, 9, 579. m/z 85 (18) O-Na+ O O (15) (13) [3] T. Aggerholm, S. C. Nanita, K. J. Koch, R. G. Cooks, J. Mass. Na Na O Na O Received: ((will be filled in by the editorial staff)) Published online on ((will be filled in by the editorial staff)) 12, 1174. H Na documented.[12,23] Our results presented here demonstrate that such reactions can be observed even in collision cells of tandem-in-space instruments when the ions show very high affinity to water (for example, ions bearing phenyl/metal bonds). O-Na+ O OH [5] V. Sabareesh, P. Balaram, Rapid Comm. Mass Spectrom. 2006, 20, 618. (16) [6] X. Tang, W. Ens, K. G. Standing, J. B. Westmore, Anal. Chem. 1988, 60, H H H Na O + H Na+ Na O 1791. + [7] D. Renner, G. Spiteller, Biomed. Environ. Mass Spectrom. 1988, 15, 75. Na O O-Na+ m/z 63 (17) O O-Na+ m/z 167 (14) Scheme 7. Proposed mechanism for the formation of m/z 63 and 85 ions The reaction of water with highly basic ions bearing twocenter-two-electron bonds between a benzene nucleus and a metal appears to be general phenomenon. For example, when the m/z 145 ion of formula Na3(C6H4)+ (which is the sodium adduct of phenelenedisodium, obtained by two consecutive losses of CO 2 from the sodium adduct of disodium terephthalate) was isolated and in an LTQ instrument at a normalized collision energy setting of 0, and a relatively long storage time (300 ms), the product ions m/z 63, m/z 85, and 123 were observed (Figure S6). The formation of these ions from m/z 145 can be rationalized by mechanisms analogous to those described in Scheme 7 for the m/z 189 ion. Thus, the peak observed at m/z 123 (Figure S6), which in fact represents the sodium adduct of phenyl sodium, originates from a rapid NaOH loss from the water adduct. Our results reveal that the ortho isomer of the disodium salts of benzene dicarboxylic acids can be distinguished from those of the meta and para isomers by the CID spectra of their sodium adducts. Although the phenomenon is clearly an ortho effect, it is dramatically different from the classical ortho effects observed under electron ionization conditions in which a small molecule is expelled from the molecular ions of some ortho compounds. In other words, in the examples discussed here, the meta and para compounds fragment differently, rather than the ortho compounds undergoing a unique pathway. Complications arising from unexpected ion-molecule reactions that take place particularly in ion traps, due to inherent long ion residence times, have been well [8] R. P. Grese, R. L. Cerny, M. L. Gross, J. Am. Chem. Soc. 1989, 111, 2835. [9] C. Zhao, B. Xie, S.-Y. Chan, C. E. Costello, P. B. O'Connor, J. Am. Soc. Mass Spectrom. 2008, 19, 138. [10] K. M. Koshy, J.Y. Wang, J. M. Boggs, Biophys. J. 1999, 77, 306. [11] M. Stano, H. D. Flosadottir, O. Ingolfsson, Rapid Comm. Mass Spectrom. 2006, 20, 3498. [12] A. B. Attygalle, N. Kharbatia, J. Bialecki, J. Ruzicka, A. Svatos, E. J. Stauber, Rapid Comm. Mass Spectrom. 2006, 20, 2265. [13] S. Bachrach, M. Hare, S. Kass, J. Am. Chem. Soc. 1998, 120, 12646. [14] G. L. Miessler, D. Tarr, Inorganic Chemistry 3rd Ed., 2004, Pearson Prentice Hall, Upper Saddle River, N.J. pp. 458. [15] D. Seyferth, Organometallics 2006, 25, 2. [16] A. A. Morton, C. E. Jr. Claff, J. Am. Chem. Soc. 1954, 76, 4935. [17] E. Baston, R. Maggi, K. Friedrich, M. Schlosser, Eur. J. Org. Chem. 2001, 21, 3985. [18] L. Pratt, S. R. Kass, J. Org. Chem. 2004, 69, 2123. [19] D. R. Reed, M. Hare, S. R. Kass, J. Am. Chem. Soc. 2000, 122, 10689. [20] G.E. Davico, V.M. Bierbaum, C.H. DePuy, G.B. Ellision, R.R. Squires, J. Am. Chem. Soc. 1995,117, 2590-2599. [21] J. B. Nicholas, B. P. Hay, J. Phys. Chem. A. 1999, 103, 9815. [22] M. Janse, W. Z. f. Mueller, Z. Naturforsch. 1989, 44, 996. 4 Entry for the Table of Contents (Please choose one layout) Layout 1: Mass Spectrometry A. Attygalle*, C.-C. Chan, F.U. Axe, M.S. Bolgar.__________ Page – Page 2.61-2.66 Å Generation of Gas-Phase Sodiated Arenes [(Na3(C6H4)+], and Other Unusual Sodium Adducts [(Na3O)+, and (Na2OH)+] Sticky Sodium: Unprecedented sodiated arenes (see structure) can be generated from sodiated disodium terephthalate and isophthalate after two successive expulsions of CO2 2.35 Å Layout 2: (Sticky Sodium) 2.61-2.66 Å ((Author(s), Corresponding Author(s)*)) __________ Page – Page Generation of Gas-Phase Sodiated Arenes [(Na3(C6H4)+], and Other Unusual Sodium Adducts [(Na3O)+, and (Na2OH)+] 2.35 Å Sticky Sodium: Unprecedented sodiated arenes (see structure) can be generated from sodiated disodium terephthalate and isophthalate after two successive expulsions of CO2 5