shift, the solution was introduced into spherical glass containers fitting into ordinary 10-mm NMR

Transcription

1 upporting Inforation for [Gd(ry-DA)(H O)] : A new MRI contrast agent for copper ion sensing Dayananda Kasala, a,b sung-heng Lin, a Chiao-Yun Chen c,d Gin-Chung Liu, c,d Chai-Lin Kao, e ian-lu Cheng, f Yun-Ming Wang a,b * *Corresponding author. E-ail address: ywang@ail.nctu.edu.tw Variable-teperature 7 O NMR easureents. For the 7 O NMR easureents, the inversion recovery ethod was applied to easure longitudinal relaxation rates, /, and the Carr-Purcell-Meiboo-Gill spin-echo technique was used to obtain transverse relaxation rates, /. In order to eliinate agnetic susceptibility corrections to cheical shift, the solution was introduced into spherical glass containers fitting into ordinary - NMR tubes. Fro the easured 7 O NMR relaxation rates and angular frequencies of the Gd(III) containing solutions, /, /, and, and of the acidified water reference, / A, / A, and A, one can calculate the reduced relaxation rates and cheical shift, / r, / r, and r (Fig. 3)(ables and ): os A r P () os A r P () os A r P (3) P is the ole fraction of bound water. / is the relaxation rates in the bound water. is the cheical shift difference between bound water and bulk water. Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry

2 A k B g B B L 3 (4) where g L is the isotropic Landé g factor (g L =. for Gd(III)) is the electron spin ( = 7/ for Gd(III)) A/ħ is the hyperfine or scalar coupling constant B is the agnetic field 3 sc A (5) where / is = / M + / ie H and are the enthalpy and entropy of activation for the exchange process: R H R h k k B ex exp (6) he 7 O longitudinal relaxation rates in Gd(III) solutions are doinated by the dipole-dipole and quadrupolar echaniss, and are given by below : R d d d GdO I I I I r (7) where I is the nuclear gyroagnetic ratio ( I = rad s for 7 O), GdO is the ean Gd(III)O distance, I is the nuclear spin (I = 5/ for 7 O), is the quadrupolar coupling constant, an asyetry paraeter, and di = M + ie + R. Using the quadrupolar coupling constant for Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry

3 Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry acidified water, ( + /3 ) / = 7.58 MHz, and estiating r =.5 Å is based on [Gd(DPA)] crystal structure., 98 E R exp (8) R R R 98.5 E R : activation energy he outer-sphere contribution to r has a siilar teperature dependence to and is given by below, where C os is an epirical constant. os = C os (9) Data analysis. he siultaneous least-squares fitting of 7 O NMR data was deterined by fitting the experiental data using the progra CIENI for WINDOW M by MICROMAH, version.. 3

4 Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry Conditional binding constant (K d ) Fluorescence spectra of [Gd(ry-DA)(H O)] ( M) in acetate buffer, ph 5.5, with excitation at 8 n show an eission axiu at 36 n that decreases upon titration of CuCl. he fluorescence was quenched copletely after addition of. equivalent as depicted in Figure 9. he conditional binding constant was obtained fro equation which relates fluorescence intensity to the concentration of Cu(II) 3 (F F) / (F F ) = ([Cu + ] / K d ) n () where F, F and F are the relative fluorescence intensities of [Gd(ry-DA)(H O)], [Gd(ry- DA)(H O)] in the presence of a know concentration of Cu +, [Gd(ry-DA)(H O)] saturated with Cu +, respectively and n is the equivalents of binding sites on [Gd(ry-DA)(H O)]. By plotting log [(F F) / (F F )] vs log [Cu + ], the slope of line is equal to n and log (K d ) is equal to log (Cu + ) where log [(F F) / (F F )] =. Figure 9 shows a typical plot of log [(F F) / (F F )] vs log [Cu + ] for [Gd(ry-DA)(H O)], which gives an apparent K d for [Gd(ry-DA)(H O)] - Cu(II) coplex of.5 M. References. M. R. pirlet, J. Rebizant, J. F. Desreux, M. F. Loncin, Inorg. Che. 984, 3, J. J. tezowski, J. L. Hoard, Isr. J. Che. 984, 4, K. C. Ciesienski, L. Y. Hyan,. Derisavifard, K. J. Franz, Inorg. Che.,, 49,

5 Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry able Luinescence lifeties of [Eu(ry-DA)(H O)] and [Eu(ry-ac-DOA)(H O)] in the presence and absence of Cu (II) at roo teperature. Coplexes τ HO /s τ DO /s q a [Eu(ry-DA)(H O)] [Eu(ry-DA)(H O)] + Cu(II) [Eu(ry-ac-DOA)(H O)] [Eu(ry-ac-DOA)(H O)] + Cu(II) a q=.(k HO -k DO -.5) 5

6 Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry able. eperature dependence of reduced transverse and longitudinal 7 O relaxation rates and reduced angular frequencies of solutions containing [Gd(ry-DA)(H O)] at 9.4. (c =. ol kg, ph = 7.4). K/ ln(/ r ) ln(/ r ) r / 6 rad s

7 Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry able 3. ignal intensity of the -weighted MR iages of [Gd(ry-DA)(H O)] and etal ions in the olar ratio = :. Concentration (M)..5.. Cu(II) ±9 53±54 985±49 73±67 La(III) 8±4 64±8 8±55 ±6 Mg(II) 3±3 36±6 6±37 6±4 Gd(III) 87±3 956± ±7 37±9 7

8 % % Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry 56test-AP 47 (3.3) C (46:5) May-8 : can E+.3e test-AP 46 (3.67) C (46:5) : can E+.9e /z Fig. EI Mass spectru of ry-da. 8

9 % Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry.6N 43 5 (.879) Cn (Cen,3, 8., Ht); (G, x.75); b (3,4. ); C (5:3-5:9x.) can E e /z Fig. EI Mass spectru of ry-ac-doa. 9

10 UV Absorbance (56 n, AU) Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry ie (in) Fig. 3 HPLC chroatogra of the [Gd(ry-DA)(H O)] coplex (PB buffer, ph = 7.4.). One peak at 3.9 in was detected by UV detector at 56 n (blue line) A upelco RP-C8 colun (5, ) was used. One peak at 3.9 in ((M EI + ): /z: 689.8[M+H] + ) was detected.

11 Relative Abundance Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry 983- #69-7 R: AV: 49 NL: 5.E6 : - c EI Full s [ ] /z Fig. 4 EI Mass spectru of [Gd(ry-DA)(H O)] coplex.

12 Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry Fig. 5 HPLC chroatogra of the [Gd(ry-ac-DOA)(H O)] coplex (PB buffer, ph = 7.4.). One peak at 9.6 in was detected by RI detector at 56 n. A upelco RP-C8 colun (5, ) was used. One peak at 9.6 in ((M EI + ): /z: 748.8[M+3H] + ) was detected.

13 % Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry ry-gd coplex (.86) Cn (Cen,3, 8., Ht); (G, x.75); b (3,4. ); C (6:7-46:55x.) can E+ 3. x 3.9e /z Fig. 6 EI Mass spectru of [Gd(ry-ac-DOA)(H O)] coplex. 3

14 F lu o re s c e n c e in te n s ity Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry [C u + ]/{[C u + ]+ [G d ( ry - D A )(H O )] - } Fig. 7 Job s plot of [Gd(ry-DA)(H O)] interaction with Cu(II) obtained by fluorescence easureents (λ ex = 8 n, λ e = 36 n). he total concentration of [Gd(ry-DA)(H O)] and Cu(II) is. M. 4

15 % Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry Gd ry Cu 79 8 (6.365) Cn (Cen,3, 8., Ht); (G, x.75); b (3,4. ); b (3,4. ); C (75:-6:7x.) can E+ 3. x.e /z Fig. 8 EI Mass spectru of [Gd(ry-DA)(H O)] coplex interaction with Cu(II). 5

16 log[( FF ) / ( FF )] Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry log[cu + ] Fig. 9 Double-logarithic plot of the quenching of fluorescence of [Gd(ry-DA)(H O)] by the titration Cu +, where F o, F and F are the relative fluorescence intensities of [Gd(ry-DA)(H O)], [Gd(ry-DA)(H O)] in the presence of a known concentration of Cu + and [Gd(ry-DA)(H O)] saturated with Cu + ( equivalents), respectively. 6

17 F lu o r e s c e n c e I n te n s ity (A U ) Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry G d (III) c o p le x : C u (II). :.. :.. :.. :.5. :.. :.. :5.. :.. : W a v e le n g th (n ) Fig. Fluorescence eission spectra of µm [Gd(ry-ac-DOA)(H O)] interaction with Cu(II), ph= 5.5 acetate buffer solution. 7

18 F lu o r e s c e n c e in te n s ity Electronic uppleentary Material (EI) for Dalton ransactions his journal is he Royal ociety of Cheistry [M n+ ]/{ [M n+ ]+ [G d (III)c o p le x ]} Fig. Quenching plots of the fluorescence of Gd(III) coplex against ole equivalents of etal ions added, Cu(II)( ), Mg(II)( ), La(III)( ) for [Gd(ry-DA)(H O)] and Cu(II) ( ) for [Gd(ry-ac- DOA)(H O)], at 37. ±. C and MHz, ph 5.5 acetate buffer solution. 8