Patrick F. James, Matthew A. Perugini and Richard A.J. O Hair

Transcription

1 Supplementary Materials for Sources of rtefacts in the Electrospray Ionization Mass Spectra of Saturated Diacylglycerophosphocholines: From ondensed Phase Hydrolysis Reactions through to Gas Phase Intercluster Reactions. Patrick F. James, Matthew. Perugini and Richard.J. O Hair List of Supplementary Figures and Tables: Figures: (S1) Positive mode ESI MS spectra of D6P in 1 mm NH4Oc, ph 6.1 as a function of concentration of D6P. (S2) ESI/MS spectra showing the effect of aging the D6P sample for 48 hrs at room temperature. (S3) LQ MS/MS spectra of the hetero-dimers. (S4) Positive mode LTQ mass spectra of 8 mm D6P solution in 1 mm NH4Oc, ph 6.1. (S5) Positive mode LQ mass spectra of D6PE in 1 mm NH4Oc, ph 6.1. (S6) Signal intensity of various ions in the positive mode LQ ESI mass spectrum of D6P as a function of tube lens voltage. (S7) spectra of (a) [D6P 2 +H-Me 3 N] + formed from MS/MS of [D6P 2 +H] + (Figure 1c) (b) [D6P 2 +H-Me 3 N] + formed via in-source ; (c) [D6P+Me] + formed from MS/MS from [D6P 2 +H] + (Figure 1c) (d) [D6P+H+Me] + formed via in-source. The mass selected precursor ion is designated by a *. (S8) spectra of (a) [D6P 2 +H- 15 H 26 O 4 ] + formed from MS/MS from [D6P 2 +H] + (Figure 1c) (b) [D6P 2 +H- 15 H 26 O 4 ] + formed via in-source. The mass selected precursor ion is designated by a *. (S9) Triple quadrupole ESI/MS spectra of D6P with increasing cone voltage. (S1) LQ spectra of authentic ion structure of to compare with intercluster reactions product (c.f. Figure 4). (S11) LTQ spectra of authentic ion structure of to compare with intercluster reactions product (c.f. Supplementary Figure S4 F). Table: (1) Errors (in ppm) associated with exact mass measurements on the LTQ- FTMS, which confirm the molecular formulas for various D6P cluster ions and products.

2 D E F 5 5 [P + H] [D6P 3 + H] [D6P 3 + H] Supplementary Figure S1: Positive mode ESI MS spectra of D6P in 1 mm NH 4 Oc, ph 6.1 as a function of concentration of D6P: (a) 8 nm; (b) 8 nm; (c) 8 nm; (d) 8 µm; (e) 8 µm; (f) 8 µm. Relevant masses are labelled, stars indicate the expected position of [D6P 2 +H] +.

3 [D6P 2 + H - R] [P + H] [P + H] H11O2-115 [D6P + Oc] [D6P + l] [D6P + 5H11O2] [D6P2+ Oc] [D6P2+ l] Intensity 1e+9 8e+8 6e+8 4e+8 2e+8 D 97 Intensity 6e+8 5e+8 4e+8 3e+8 2e+8 1e+8 E [D6P + H - R] [D6P 2 + H - R 2 ] [D6P 2 + H - R] + 89 [D6P 2 + H] Supplementary Figure S2: ESI/MS spectra showing the effect of aging the D6P sample for 48 hrs at room temperature: (a) Positive mode LQ ESI/MS of fresh solution; (b) LQ ESI/MS of aged solution; (c) Negative mode ESI/MS of aged solution; (d) Positive mode QQQ ESI/MS of fresh solution; (e) QQQ ESI/MS of aged solution.

4 [D5P + H] [D14P + H] [D5P + D14P + H] [D5P + D6P + H N(H 3 ) 3 ] + 82 [D5P + D6P + H] [D5P + H] [D5P + D14P + H N(H 3 ) 3 ] # # ^ ^ # # ^ ^ [D14P + H] Supplementary Figure S3: LQ MS/MS spectra of the hetero-dimers: (a) [D5P+H+D6P] +, (b) [D5P+H+D14P] + and (c) [D6P+H+D14P] +. # denotes methylated monomer, ^ denotes monomer + phosphocholine. 5 [D6P + D14P + H N(H 3 ) 3 ] + 82 [D6P + D14P + H] # # ^ ^

5 [D6P + H 3 ] + 97 [D6P + H - R] [D6P 2 + H - R] H 26 O 4 -N(H 3 ) [P + H] H 26 O D H 26 O 4 38 [D6P 2 + H - N(H 3 ) 3 ] [P + H 3 ] E 97 F [P + H] [P + H 3 ] [D6P + H 3 ] + [D6P 2 + H - 15 H 26 O 4 ] + [D6P + H 3 ] Supplementary Figure S4: Positive mode LTQ mass spectra of 8 mm D6P solution in 1 mm NH 4 Oc, ph 6.1: (a) Full ESI spectrum; (b) MS/MS of ; (c) MS 3 of ; (d) MS 3 of [D6P 2 + H - N(H 3 ) 3 ] + ; (e) MS 3 of [D6P + H 3 ] + ; (f) MS 3 of [D6P 2 + H - 15 H 26 O 4 ] +.

6 [D6PE 2 + H] [D6PE + H] x [D6PE + H] [D6PE 3 + H] [ 15 H 26 O 4 + H] [ 15 H 26 O 4 + H] [D6PE 4 + H] [D6PE 2 + H - 15 H 26 O 4 ] [D6PE 2 + H] [ 15 H 26 O 4 + H] [D6PE 2 + H - PE] Supplementary Figure S5: Positive mode LQ mass spectra of D6PE in 1 mm NH 4 Oc, ph 6.1: (a) Full ESI MS spectrum; (b) MS/MS of [D6PE 2 + H] + ; (c) MS/MS of [D6PE + H] +. 5 [D6PE + H]

7 (arb) 8 6 (arb) Tube Lens Voltage (V) Tube Lens Voltage (V) D 5 (arb) (arb) Tube Lens Voltage (V) Tube Lens Voltage (V) E (arb) Tube Lens Voltage (V) Supplementary Figure S6: Signal intensity of various ions in the positive mode LQ ESI mass spectrum of D6P as a function of tube lens voltage: (a) [D6P 2 +H] + ( 97); (b) [D6P+H] + ( ); (c) [D6P 2 +H- N(H3) 3 ] + ( ); (d) [D6P 2 +H 3 ] + ( ); (e) [D6P 2 +H- 15 H 26 O 4 ] + ( ).

8 H 26 O H 26 O [D6P2 + H -Me 3 N] s D - 215H 26 O H 26 O [D6P 2+ H -Me3N] [P + H 3 ] [D6P + H + H3] [P + H3] [D6P + H + H 3] + s Supplementary Figure S7: spectra of (a) [D6P2+H-Me3N]+ formed from MS/MS of [D6P2+H]+ (Figure 1c) (b) [D6P2+H-Me3N]+ formed via in-source ; (c) [D6P+Me]+ formed from MS/MS from [D6P2+H]+ (Figure 1c) (d) [D6P+H+Me]+ formed via in-source. The mass selected precursor ion is designated by a *.

9 5 [D6P + H + H 3] [D6P + H + H3] + s Supplementary Figure S8: spectra of (a) [D6P2+H-15H26O4]+ formed from MS/MS from [D6P2+H]+ (Figure 1c) (b) [D6P2+H-15H26O4]+ formed via in-source. The mass selected precursor ion is designated by a *.

10 1 V V 15 V Supplementary Figure S9: Triple quadrupole ESI/MS spectra of D6P with increasing cone voltage (a) [D6P+H] +, cone voltage = 1 V; (b) [D6P+H] +, cone voltage = V; (c) [D6P+H] +, cone voltage = 15 V. ^ denotes monomer + phosphocholine.

11 [D6P 2 + H - N(H 3 ) 3 ] + [D6P 2 + H + P] + 5 [D6P 2 + H + P] + [D6P 2 + H + P] + 19 [D6P 2 + H 3 ] Supplementary Figure S1: LQ spectra of authentic ion structure of to compare with intercluster reactions product (c.f. Figure 4): (a) MS/MS of hetero trimer [D6P 2 + H + P] + ; (b) MS 3 of heterodimer [D6P + H + P] +.

12 [D6P 2 + H + P] + 5 [P + H] [D6P 2 + H 3 ] + [D6P 2 + H 3 ] + [D6P 2 + H - N(H 3 ) 3 ] + [D6P 2 + H + P] [P + H 3 ] [D6P 2 + H + P] Supplementary Figure S11: LTQ spectra of authentic ion structure of to compare with intercluster reactions product (c.f. Supplementary Figure S4 F): (a) MS/MS of hetero trimer [D6P 2 + H + P] + ; (b) MS 3 of heterodimer [D6P + H + P] +.

13 Supplementary Table S1: Errors (in ppm) associated with exact mass measurements on the LTQ-FTMS, which confirm the molecular formulas for various D6P cluster ions and products. Species hemical Formula ppm (source) ppm () 4 H 81 O 16 N 2 P [D6P 1 + H] + 2 H 41 O 8 N 1 P fragmentation S n 2 X-linked 37 H 72 O 16 N 1 P D6P + Me 21 H 43 O 8 N 1 P E 2 25 H 55 O 12 N 2 P S n 2 X-linked fragmentation S n 2 Glycerocyl 22 H 46 O 12 N 1 P S n 2 Glycerocyl 2 7 H 2 O 8 N 1 P E2 fragmentation [D6P 1 + H] + 2 H 41 O 8 N 1 P D6P + Me 21 H 43 O 8 N 1 P P 5 H 15 O 4 N 1 P D6P + Me fragmentation P + Me 6 H 17 O 4 N 1 P [D6P 2 + Na] + 4 H 8 O 16 N 2 P 23 2 Na [D6P 1 + Na] + 2 H 4 O 8 N 1 P 23 1 Na [D6P 2 + K] + 4 H 8 O 16 N 2 P 39 2 K [D6P 1 + K] + 2 H 4 O 8 N 1 P 39 1 K 1.64