Supporting information for: Red-Emitting Ruthenium(II) and Iridium(III) Complexes as Phosphorescent Probes for Methylglyoxal in vitro and in vivo Wenzhu Zhang, *, Feiyue Zhang, Yong-Lei Wang, Bo Song, Run Zhang, *, Jingli Yuan State Key Laboratory of Fine Chemicals, School of Chemistry, Dalian University of Technology, Dalian 116024, PR China Email: wzhzhang@dlut.edu.cn (W. Zhang), Tel./Fax: +86 41184986041. Applied Physical Chemistry, Department of Chemistry, KTH Royal Institute of Technology, SE- 100 44 Stockholm, Sweden. Australian Institute for Bioengineering and anotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia Email: r.zhang@uq.edu.au (R. Zhang), Tel: + 61 7 3346 3806, Fax: + 61 7 3346 3978
General information for sample preparation Hydrogen peroxide (H 2 O 2 ) was diluted immediately from a stabilized 30% solution, and was assayed using its molar absorption coefficient of 43.6 M 1 cm 1 at 240 nm. 1, 2 Singlet oxygen ( 1 O 2 ) was generated from the a 2 MoO 4 -H 2 O 2 system in 0.05 M carbonate buffer of ph 10.5. 3 A stock solution of HOCl was prepared by dilution of the commercial sodium hypochlorite solution and stored according to the previous literatures. The concentration of HOCl was determined by using its molar extinction coefficient of 391 M 1 cm 1 at 292 nm before usage. 4 OOO - was donated by 3- morpholinosydnonimine. Hydroxylradical ( OH) was generated in the Fenton system from ferrous ammonium sulfate and hydrogen peroxide. itric oxide (O) aqueous solution was prepared by passing O gas through a deoxidized PBS buffer for 3 h, and the concentration was measured by using reported method. 5 Metal ions, including a +, K +, Mg 2+, Ca 2+, Zn 2+, Mg 2+, Fe 2+, Fe 3+, Cu 2+, and i 2+ were daily prepared as nitrate salts in DI water. Cell line and cell culture RAW 264.7 macrophage cells were obtained from Dalian Medicine University. The cells were cultured in Dulbecco s modified Eagle s medium (DMEM, Sigma-Aldrich, Inc.), supplemented with 10% fetal bovine serum (FBS), 1% penicillin, 1% streptomycin sulphate in a humidified 5% CO 2 /95% air incubator at 37 o C. The growth medium was changed every two days. The cells were routinely subcultured using 0.05% trypsin-edta solution and growth to 80% confluence prior to experiments. Cytotoxicity analysis The cytotoxicity of the complexes towards RAW 264.7 macrophage cells was examined by MTT analysis, which involves the reduction of [3-(4,5-dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide] tetrazolium to an insoluble formazan crystal by the metabolic activity of living cells. 6 RAW 264.7 macrophage cells were seeded at a density of 5 10 4 cells/ml in a 96-well micro-assay
culture plate and growth for 24 h at 37 o C in a 5% CO 2 /95% air incubator. Then, [Ru(bpy) 2 (DAphen)](PF 6 ) 2 or [Ir(ppy) 2 (DA-phen)](PF 6 ) in fresh culture medium was added into each well with different final concentrations of 0, 25, 50, 100, 150, 200 µm. Control wells were prepared by the addition of culture medium, and wells containing culture media without cells were used as blanks. After incubation at 37 o C in a 5% CO 2 /95% air incubator for 24 h, cell culture medium was removed and cells were washed three times with PBS. Then, 100 µl of 0.5 mg/ml MTT solution in PBS was added to each well, and the cells were incubated for another 4 h. The excess MTT solution was then carefully removed from each well, and the formed formazan was dissolved in 100 µl of DMSO (dimethyl sulfoxide). The optical density of each well was measured at a wavelength of 570 nm referenced at 690 nm using a microplate reader (Bio-Rad, xmark). The results from the five individual experiments were averaged. The following formula was used to calculate the viability of cell growth: Vialibity(%) = (mean of absorbance value of treatment group blank)/(mean absorbance value of control blank) 100. Ir Cl Cl Ir + H 2 H 2 (a)ch 2 Cl 2 /MeOH(1:2);reflux;4h (b)h 4 PF 6 ;1h Ir H 2 H 2 PF 6 Scheme S1. Synthetic pathway of [Ir(ppy) 2 (DA-phen)]PF 6.
Figure S1. 1 H MR spectrum of [Ir(ppy) 2 (DA-phen)]PF 6. Figure S2. 13 C MR spectrum of [Ir(ppy) 2 (DA-phen)]PF 6.
Figure S3. ESI-HRMS of [Ir(ppy) 2 (DA-phen)]PF 6. Figure S4. ITMS of [Ru(bpy) 2 (DA-phen)](PF 6 ) 2 reacted with MGO.
Figure S5. ITMS of [Ir(ppy) 2 (DA-phen)](PF 6 ) reacted with MGO. 750 500 Current (µa) 250 0-250 -500-2 -1 0 1 2 Potential (V) vs Ag/AgO 3 Figure S6. Cyclic voltammograms of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) (the product of [Ru(bpy) 2 (DA-phen)] 2+ reacted with MGO) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s.
400 300 Current (µa) 200 100 0-100 -200-2 -1 0 1 2 Potential (V) vs Ag/AgO 3 Figure S7. Cyclic voltammograms of 1.0 mm [Ir(ppy) 2 (DA-phen)] + (dash line) and [Ir(ppy) 2 (MPphen)] + (solid line) (the product of [Ir(ppy) 2 (DA-phen)] + reacted with MGO) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s. 150 Current (µa) 100 50 0-50 -100-2 -1 0 1 2 Potential (V) vs Ag/AgCl Figure S8. Cyclic voltammograms of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) (the product of [Ru(bpy) 2 (DA-phen)] 2+ reacted with MGO) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s.
120 80 Current (µa) 40 0-40 -80-2 -1 0 1 2 Potential (V) vs Ag/AgCl Figure S9. Cyclic voltammograms of 1.0 mm [Ir(ppy) 2 (DA-phen)] + (dash line) and [Ir(ppy) 2 (MPphen)] + (solid line) (the product of [Ir(ppy) 2 (DA-phen)] + reacted with MGO) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s. 200 160 Current (µa) 120 80 40 0 0.0 0.5 1.0 1.5 Potential (V) vs Ag/AgCl Figure S10. Differential pulse voltammetry of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s.
15 Current (µa) 10 5 0 0.0 0.5 1.0 1.5 Potential (V) vs Ag/AgCl Figure S11. Differential pulse voltammetry of 1.0 mm [Ir(bpy) 2 (DA-phen)] + (dash line) and [Ir(bpy) 2 (MP-phen)] + (solid line) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s. 60 Current (µa) 40 20 0 0.0 0.3 0.6 0.9 1.2 Potential (V) vs Ag/AgO 3 Figure S12. Differential pulse voltammetry of 1.0 mm [Ir(bpy) 2 (DA-phen)] + (dash line) and [Ir(bpy) 2 (MP-phen)] + (solid line) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s.
20 15 Current (µa) 10 5 0 0.0 0.3 0.6 0.9 1.2 Potential (V) vs Ag/AgO 3 Figure S13. Differential pulse voltammetry of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s. 12000 ECL intensity (a.u.) 10000 8000 6000 4000 2000 0 7.5 10.0 Scan time (s) Figure S14. ECL behaviors of [Ru(bpy) 2 (DA-phen)] 2+ (100 µm) in the absence (dash line) and presence (solid line) of MGO (800 µm) in 50 mm PBS buffer of ph 7.4 containing 10 mm TPrA at room temperature. Scan rate of ECL was 0.15 v/s.
2000 ECL intensity (a.u.) 1500 1000 500 0 7.5 10.0 Scan time (s) Figure S15. ECL behaviors of [Ir(bpy) 2 (DA-phen)] + (100 µm) in the absence (dash line) and presence (solid line) of MGO (800 µm) in 50 mm PBS buffer of ph 7.4 containing 10 mm TPrA at room temperature. Scan rate of ECL was 0.15 v/s. Luminescence intensity (a.u.) 250 A 200 150 100 50 4 6 8 10 ph Luminescence intensity (a.u.) 90 80 70 60 50 40 30 20 10 0 B 4 6 8 10 ph Figure S16. Effects of ph on the phosphorescence responses of [Ru(bpy) 2 (DA-phen)] 2+ and [Ir(ppy) 2 (DA-phen)] + towards MGO. Phosphorescence intensities of [Ru(bpy) 2 (DA-phen)] 2+ (A, 10 µm in 25 mm PBS buffer) and [Ir(ppy) 2 (DA-phen)] + (B, 10 µm in 2:1 EtOH-25mM PBS buffer) at different ph values in the absence (, black line) and presence (, red line) of 800 µm MGO.
Intensity (a. u.) 200 100 0 A a b c d PL Enhancement factor 18 15 12 9 6 3 0 B a b c d e f Figure S17. (A) Phosphorescence intensity of (a) [Ru(bpy) 2 (DA-phen)](PF 6 ) 2 (10 µm) in PBS buffer, (b) [Ru(bpy) 2 (MP-phen)](PF 6 ) 2 (10 µm) in 25 mm PBS buffer of ph 7.4, (c) [Ir(ppy) 2 (DAphen)](PF 6 ) (10 µm) in PBS-EtOH (1:2, v/v)and (d) [Ir(ppy) 2 (MP-phen)](PF 6 ) (10 µm) in PBS- EtOH (1:2, v/v) in the absence (grey bars) and presence (blank bars) of calf thymus DA; (B) changes in phosphorescence intensity of the probes and their reaction products with MGO in the absence (grey bars) and presence (black bars) of calf thymus DA. (a) [Ru(bpy) 2 (DA-phen)](PF 6 ) 2 (10 µm) in PBS buffer; (b) [Ru(bpy) 2 (MP-phen)](PF 6 ) 2 (10 µm) in 25 mm PBS buffer of ph 7.4; (c) [Ir(ppy) 2 (DA-phen)](PF 6 ) (10 µm) in PBS; (d) [Ir(ppy) 2 (MP-phen)](PF 6 ) (10 µm) in 25 mm PBS buffer of ph 7.4; (e) [Ir(ppy) 2 (DA-phen)](PF 6 ) (10 µm) in PBS-EtOH (1:2, v/v); (f) [Ir(ppy) 2 (MPphen)](PF 6 ) (10 µm) in PBS-EtOH (1:2, v/v).
Ru H 2 H 2 [Ru(bpy) 2 (DA-phen)] 2+ Figure S18. Optimized molecular geometry of [Ru(bpy) 2 (DA-phen)] 2+ in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S1. Cartesian coordinates of [Ru(bpy) 2 (DA-phen)] 2+ in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C -1.572254 2.692582-0.737684 2 C 0.059661 2.007338-2.253476 3 C -0.032512 3.194707-2.964496 4 C -0.93408 4.1638-2.534742 5 C -1.709162 3.907572-1.411713 6 C -2.355466 2.328407 0.45814 7 C -2.752499 0.705601 2.081392 8 C -3.731325 1.479222 2.687322 9 C -4.02487 2.728038 2.148278 10 C -3.329695 3.152758 1.024047 11 H 0.746615 1.22285-2.550231 12 H 0.595301 3.348781-3.836341 13 H -1.03412 5.106649-3.064403 14 H -2.414842 4.653619-1.065027 15 H -2.489436-0.271151 2.471827 16 H -4.247258 1.102087 3.564541 17 H -4.784115 3.363751 2.594221 18 H -3.548948 4.121855 0.591375 19 C -2.354776-2.32897-0.457911 20 C -3.328804-3.153644-1.023693 21 C -4.024305-2.729135-2.1478 22 C -3.731285-1.480197-2.68685
23 C -2.752632-0.706253-2.081052 24 C -1.57127-2.692904 0.737792 25 C -1.707673-3.907951 1.411823 26 C -0.932355-4.163929 2.534747 27 C -0.031071-3.194527 2.964402 28 C 0.0606-2.007118 2.253386 29 H -3.547647-4.122832-0.591018 30 H -4.783397-3.365101-2.593643 31 H -4.247493-1.103216-3.563972 32 H -2.489972 0.270601-2.471499 33 H -2.413141-4.654237 1.065221 34 H -1.031999-5.106821 3.064405 35 H 0.596906-3.348395 3.836165 36 H 0.747318-1.222396 2.550067 37 C 2.296265-0.459199-0.53384 38 C 1.040458-1.748232-2.040492 39 C 2.17599-2.281605-2.616292 40 C 3.439405-1.892446-2.122731 41 C 3.509588-0.978693-1.079816 42 C 2.296231 0.459737 0.533594 43 C 1.040297 1.748865 2.040051 44 C 2.175782 2.282502 2.615723 45 C 3.439223 1.893362 2.122249 46 C 3.509511 0.979389 1.079532 47 H 0.054469-2.028709-2.398339 48 H 2.088764-2.987694-3.434769 49 H 0.054283 2.029285 2.397874 50 H 2.088484 2.988758 3.434049 51 H -0.68739 1.751628-1.16564 52 Ru -0.574052 0.000003-0.000009 53-2.072792-1.110403-0.994108 54 1.064445 0.859202 1.026797 55 1.064518-0.858745-1.027063 56-2.072975 1.109954 0.994327 57-0.686671-1.751653 1.165644 58 4.329173 2.312321 2.581854 59 C 4.753403-0.5314-0.481215 60 C 4.753382 0.531974 0.481147 61 5.924048-1.07705-0.848643 62 H 5.95252-1.838131-1.513451 63 H 6.755836-0.982009-0.280328 64 5.92404 1.077435 0.848812 65 H 5.952542 1.838348 1.513811 66 H 6.755907 0.982368 0.280617 67 H 4.329401-2.311189-2.582442
Ru [Ru(bpy) 2 (DA-phen)] 2+ Figure S19. Optimized molecular geometry of [Ru(bpy) 2 (MP-phen)] 2+ in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S2. Cartesian coordinates of [Ru(bpy) 2 (MP-phen)] 2+ in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C -1.872081 2.742029-0.642907 2 C -0.23463 2.012739-2.209981 3 C -0.26032 3.211363-2.878533 4 C -1.134144 4.229847-2.409265 5 C -1.918746 3.995738-1.312381 6 C -2.620834 2.401897 0.508254 7 C -3.098941 0.707566 2.113921 8 C -4.041042 1.486562 2.737173 9 C -4.287454 2.790698 2.225942 10 C -3.592062 3.231792 1.132685 11 H 0.410195 1.204231-2.537698 12 H 0.373427 3.360032-3.746115 13 H -1.17513 5.190984-2.914254 14 H -2.579006 4.772946-0.942215 15 H -2.871802-0.290723 2.472599 16 H -4.572529 1.104148 3.601872 17 H -5.029073 3.431021 2.695449 18 H -3.785858 4.219994 0.729195 19 C -2.985802-2.148103-0.428536 20 C -4.066414-2.843102-0.96775 21 C -4.783702-2.28793-2.020429 22 C -4.411485-1.042748-2.514995
23 C -3.330473-0.393441-1.938863 24 C -2.167911-2.641227 0.694318 25 C -2.368076-3.871417 1.317819 26 C -1.545227-4.247027 2.3724 27 C -0.534721-3.385575 2.784634 28 C -0.380355-2.176268 2.122711 29 H -4.352666-3.810238-0.571913 30 H -5.626353-2.824212-2.446213 31 H -4.944542-0.571543-3.333871 32 H -2.998605 0.577261-2.287583 33 H -3.158005-4.535506 0.988118 34 H -1.694894-5.203104 2.864831 35 H 0.129016-3.636729 3.605113 36 H 0.391343-1.472771 2.41433 37 C 1.84896-0.617019-0.602689 38 C 0.511747-1.909893-1.992349 39 C 1.636079-2.493259-2.583627 40 C 2.897013-2.115934-2.165421 41 C 3.025063-1.15432-1.149213 42 C 1.910308 0.365008 0.449584 43 C 0.734584 1.747054 1.906322 44 C 1.924533 2.241444 2.446002 45 C 3.133815 1.778109 1.965021 46 C 3.144973 0.816645 0.941024 47 H -0.489641-2.181472-2.308304 48 H 1.499406-3.232673-3.365605 49 H -0.232271 2.085957 2.260773 50 H 1.878856 2.983599 3.235964 51-1.016577 1.750237-1.135276 52 Ru -0.99218 0.012307-0.000975 53-2.6307-0.933293-0.926975 54 0.726548 0.829368 0.936787 55 0.611874-0.991671-1.029939 56-1.872081 2.742029-0.642907 57-0.23463 2.012739-2.209981 58 H -0.26032 3.211363-2.878533 59 C -1.134144 4.229847-2.409265 60 C -1.918746 3.995738-1.312381 61-2.620834 2.401897 0.508254 62-3.098941 0.707566 2.113921 63 H -4.041042 1.486562 2.737173 64 C -4.287454 2.790698 2.225942 65 C -3.592062 3.231792 1.132685 66 H 0.410195 1.204231-2.537698 67 H 0.373427 3.360032-3.746115 68 H -1.17513 5.190984-2.914254 69 C -2.579006 4.772946-0.942215 70 H -2.871802-0.290723 2.472599
Ir H 2 H 2 [Ir(ppy) 2 (DA-phen)] + Figure S20. Optimized molecular geometry of [Ir(ppy) 2 (DA-phen)] + in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S3. Cartesian coordinates of [Ir(ppy) 2 (DA-phen)] + in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C -1.586859 2.627225-0.684778 2 C 0.165879 2.063525-2.17499 3 C -0.006493 3.218909-2.902712 4 C -1.039127 4.108517-2.522085 5 C -1.813679 3.810578-1.427345 6 C -2.291007 2.251031 0.497497 7 C -2.552034 0.598892 2.175726 8 C -3.511773 1.336508 2.830353 9 C -3.871314 2.599253 2.30137 10 C -3.264972 3.043378 1.152079 11 H 0.92649 1.338914-2.444193 12 H 0.636673 3.421422-3.752513 13 H -1.220291 5.017555-3.088906 14 H -2.609549 4.484605-1.12816 15 H -2.235897-0.369574 2.546555 16 H -3.972004 0.942624 3.730271 17 H -4.614358 3.21487 2.800749 18 H -3.523779 4.014992 0.744338 19 C -2.342631-2.202076-0.523174
20 C -3.327206-2.969043-1.191357 21 C -3.903379-2.510504-2.350753 22 C -3.501629-1.259342-2.875755 23 C -2.532326-0.546739-2.207163 24 C -1.665901-2.595865 0.670083 25 C -1.935748-3.771421 1.409829 26 C -1.18333-4.090771 2.514284 27 C -0.130357-3.231215 2.906731 28 C 0.083466-2.08124 2.181165 29 H -3.618522-3.932651-0.786584 30 H -4.654946-3.106694-2.860822 31 H -3.937035-0.854796-3.783244 32 H -2.183901 0.41199-2.574218 33 H -2.746622-4.422806 1.100987 34 H -1.397308-4.993856 3.079139 35 H 0.497007-3.451989 3.763764 36 H 0.861487-1.378858 2.459311 37 C 2.343862-0.451325-0.547678 38 C 1.065176-1.679945-2.072389 39 C 2.220674-2.186394-2.665723 40 C 3.468056-1.807352-2.189597 41 C 3.566358-0.911869-1.107525 42 C 2.345675 0.408038 0.570007 43 C 1.07549 1.609929 2.123703 44 C 2.232076 2.068526 2.750841 45 C 3.478133 1.684839 2.271187 46 C 3.566278 0.845136 1.146224 47 H 0.077931-1.954704-2.425443 48 H 2.128133-2.881385-3.493809 49 H 0.08955 1.891078 2.475775 50 H 2.142644 2.722383 3.612028 51 H -0.597507 1.746094-1.106038 52 Ir -0.486986 0.000639 0.000394 53-1.952981-0.98383-1.067157 54 1.121796 0.794382 1.065308 55 1.117952-0.833503-1.040064 56-1.942948 1.021473 1.04545 57-0.657954-1.742613 1.103522 58 4.369239 2.032359 2.784144 59 C 4.802742-0.4498-0.538782 60 C 4.803189 0.42946 0.537164 61 6.04802-0.815822-1.073726 62 H 6.011595-1.438883-1.870256 63 H 6.696248-1.184315-0.381723 64 6.023443 0.855411 1.053373 65 H 5.997665 1.728629 1.566027 66 H 6.760915 0.876295 0.353819 67 H 4.354068-2.220603-2.660695
Ir [Ir(ppy) 2 (MP-phen)] + Figure S21. Optimized molecular geometry of [Ir(ppy) 2 (MP-phen)] + in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S4. Cartesian coordinates of [Ir(ppy) 2 (MP-phen)] + in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C -1.859734 2.66479-0.768604 2 C -0.292545 1.8755-2.348589 3 C -0.422072 3.009266-3.121804 4 C -1.328411 4.006651-2.709368 5 C -2.037976 3.830103-1.542701 6 C -2.535847 2.389711 0.469238 7 C -2.908644 0.797482 2.172487 8 C -3.787019 1.630673 2.831469 9 C -4.030516 2.912118 2.297596 10 C -3.407136 3.282371 1.127087 11 H 0.377588 1.072399-2.632972 12 H 0.165644 3.111784-4.027752 13 H -1.470302 4.904321-3.304383 14 H -2.743333 4.587727-1.219077 15 H -2.674071-0.189156 2.555201 16 H -4.266469 1.292228 3.743762 17 H -4.699784 3.602896 2.8024 18 H -3.581724 4.268016 0.70968 19 C -2.749994-2.234478-0.357043 20 C -3.728694-3.05186-0.958738 21 C -4.385335-2.63183-2.093952 22 C -4.067108-1.375873-2.647982
23 C -3.086211-0.618131-2.044671 24 C -2.026994-2.565188 0.840464 25 C -2.255114-3.711942 1.628557 26 C -1.49222-3.949247 2.750107 27 C -0.4813-3.032648 3.101637 28 C -0.305158-1.912331 2.317872 29 H -3.960146-4.019381-0.526797 30 H -5.13742-3.264957-2.555934 31 H -4.56798-0.999406-3.533478 32 H -2.792337 0.345133-2.445447 33 H -3.040699-4.406654 1.352067 34 H -1.672309-4.832446 3.356312 35 H 0.150853-3.185866 3.969735 36 H 0.446533-1.169012 2.557338 37 C 1.894362-0.555387-0.619045 38 C 0.529062-1.819288-2.051646 39 C 1.635392-2.366999-2.679396 40 C 2.923685-1.989114-2.267162 41 C 3.065101-1.079673-1.227766 42 C 1.962307 0.360705 0.446079 43 C 0.785334 1.704435 1.970106 44 C 1.965735 2.170577 2.525639 45 C 3.194267 1.710191 2.025681 46 C 3.203899 0.801305 0.976338 47 H -0.477787-2.087325-2.352626 48 H 1.489574-3.083091-3.481127 49 H -0.178934 2.037659 2.337963 50 H 1.922457 2.887757 3.338384 51-0.98817 1.685538-1.207793 52 Ir -0.895297 0.006374 0.003122 53-2.428031-1.021842-0.937932 54 0.759709 0.816455 0.961034 55 0.631538-0.926908-1.050356 56-2.283047 1.151167 1.030125 57-1.051584-1.662896 1.221607 58 H 4.134603 2.055733 2.442564 59 C 4.355713-0.634999-0.709409 60 C 4.426051 0.276182 0.371993 61 5.480781-1.111651-1.272556 62 5.611221 0.681993 0.859356 63 H 3.807576-2.399797-2.744135 64 C 6.720488 0.207619 0.300005 65 C 8.042061 0.665769 0.847432 66 H 8.115177 0.427575 1.915041 67 H 8.87909 0.192939 0.325716 68 H 8.138746 1.753509 0.747655 69 C 6.63553-0.697751-0.777611 70 H 7.54066-1.088635-1.240668
Figure S22. TDDFT-based calculated absorption spectrum of [Ru(bpy) 2 (DA-phen)] 2+. Figure S23. TDDFT-based calculated absorption spectrum of [Ru(bpy) 2 (MP-phen)] 2+.
Figure S24. TDDFT-based calculated absorption spectrum of [Ir(ppy) 2 (DA-phen)] +. Figure S25. TDDFT-based calculated absorption spectrum of [Ir(ppy) 2 (MP-phen)] +.
Table S5. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ru(bpy) 2 (DA-phen)] 2+. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ru(bpy) 2 (D Electronic TDDFT// B3LYP//6-311*G(d) A-phen)] 2+ transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T 1 1.39 0.00 HOMO LUMO 0.63 L LCT HOMO LUMO+1 0.31 L LCT/L MCT S 0 T 2 1.75 0.00 HOMO LUMO+3 0.68 IL CT/L MCT S 0 T 3 1.76 0.00 HOMO LUMO 0.66 L LCT HOMO LUMO+1 0.26 L LCT/L MCT S 0 T 4 2.06 0.00 HOMO LUMO 0.31 L LCT HOMO LUMO+1 0.63 L LCT/L MCT S 0 T 5 2.07 0.00 HOMO LUMO 0.26 L LCT HOMO LUMO+1 0.66 L LCT/L MCT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: bpy, L : DA-phen. Figure S26. Representative frontier molelcular orbital distributions of [Ru(bpy) 2 (DA-phen)] 2+ (isodensity contour = 0.02 a.u.).
Table S6. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ru(bpy) 2 (MP-phen)] 2+. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ru(bpy) 2 (M Electronic TDDFT// B3LYP//6-311*G(d) P-phen)] 2+ transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T 1 2.01 0.00 HOMO-4 LUMO 0.14 L LCT/MLCT HOMO-3 LUMO 0.13 L LCT/MLCT HOMO LUMO 0.65 MLCT HOMO LUMO+1 0.10 LL CT/ML CT S 0 T 2 2.27 0.00 HOMO LUMO 0.69 MLCT S 0 T 3 2.29 0.00 HOMO-1 LUMO 0.55 MLCT HOMO-2 LUMO 0.38 MLCT HOMO-1 LUMO+1 0.13 MLCT/ML CT S 0 T 4 2.45 0.00 HOMO-3 LUMO 0.10 LLCT/L LCT HOMO-2 LUMO 0.55 MLCT HOMO-1 LUMO 0.37 MLCT HOMO LUMO 0.11 MLCT S 0 T 5 2.54 0.00 HOMO LUMO+1 0.34 LL CT/ML CT HOMO LUMO+2 0.50 MLCT HOMO LUMO+3 0.21 ML CT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: bpy, L : MP-phen. Figure S27. Representative frontier molelcular orbital distributions of [Ru(bpy)2(MP-phen)]2+ (isodensity contour = 0.02 a.u.).
Table S7. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ir(ppy) 2 (DA-phen)] +. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ir(ppy) 2 (DAphen)] Electronic TDDFT// B3LYP//6-311*G(d) + transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T 1 1.31 0.00 HOMO LUMO 0.24 L LCT/IL CT S 0 T 2 1.46 0.00 HOMO LUMO+1 0.78 L LCT/IL CT S 0 T 3 1.69 0.00 HOMO LUMO+2 0.71 L LCT S 0 T 4 1.86 0.00 HOMO LUMO+3 0.71 IL CT S 0 T 5 1.94 0.00 HOMO LUMO+4 0.66 L LCT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: ppy, L : DA-phen. Figure S28. Representative frontier molelcular orbital distributions of [Ir(ppy) 2 (DA-phen)] + (isodensity contour = 0.02 a.u.).
Table S8. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ir(ppy) 2 (MP-phen)] +. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ir(ppy) 2 (MPphen)] Electronic TDDFT// B3LYP//6-311*G(d) + transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T 1 1.81 0.00 HOMO LUMO 0.64 LL CT/LMCT HOMO LUMO+1 0.52 L LCT/L MCT S 0 T 2 1.86 0.00 HOMO LUMO 0.63 LL CT/LMCT HOMO LUMO+1 0.47 L LCT/L MCT S 0 T 3 1.91 0.00 HOMO LUMO+2 0.66 LL CT HOMO LUMO+3 0.25 LL CT S 0 T 4 1.99 0.00 HOMO LUMO+2 0.71 LL CT HOMO LUMO+3 0.25 LL CT S 0 T 5 2.05 0.00 HOMO LUMO+4 0.70 L LCT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: ppy, L : MP-phen. Figure S29. Representative frontier molelcular orbital distributions of [Ir(ppy) 2 (MP-phen)] + (isodensity contour = 0.02 a.u.).
References 1. Miller, E. W.; Albers, A. E.; Pralle, A.; Isacoff, E. Y.; Chang, C. J., Boronate-Based Fluorescent Probes for Imaging Cellular Hydrogen Peroxide. J. Am. Chem. Soc.2005,127, 16652-16659. 2. Yuan, L.; Lin, W.; Xie, Y.; Chen, B.; Zhu, S., Single Fluorescent Probe Responds to H2O2, O, and H2O2/O with Three Different Sets of Fluorescence Signals. J. Am. Chem. Soc.2012,134, 1305-1315. 3. Ye, Z.; Song, B.; Yin, Y.; Zhang, R.; Yuan, J., Development of singlet oxygen-responsive phosphorescent ruthenium(ii) complexes. Dalton Trans.2013,42, 14380-14383. 4. Chen, X.; Wang, X.; Wang, S.; Shi, W.; Wang, K.; Ma, H., A Highly Selective and Sensitive Fluorescence Probe for the Hypochlorite Anion. Chem. Eur. J.2008,14, 4719-4724. 5. Zhang, R.; Ye, Z.; Wang, G.; Zhang, W.; Yuan, J., Development of a Ruthenium(II) Complex Based Luminescent Probe for Imaging itric Oxide Production in Living Cells. Chem. Eur. J.2010,16, 6884-6891. 6. Li, C.; Yu, M.; Sun, Y.; Wu, Y.; Huang, C.; Li, F., A onemissive Iridium(III) Complex That Specifically Lights-Up the uclei of Living Cells. J. Am. Chem. Soc.2011,133, 11231-11239.