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REDA- · PDF file Fig. S1 1H NMR spectra of REDA-2PC (black), REDA-2PC + 0.5 equivalent of NCS– (red), REDA-2PC + 1 equivalent of NCS– (blue), REDA-2PC + 2.5 equivalents of NCS–

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Text of REDA- · PDF file Fig. S1 1H NMR spectra of REDA-2PC (black), REDA-2PC + 0.5 equivalent of...

  • Electronic Supplementary Information (ESI)

    A smart rhodamine-pyridine conjugate for bioimaging of thiocyanate in living

    cells cells Sandip Mandal,a Animesh Sahana,a Arnab Banerjee,a Damir A. Safin,*b Maria G. Babashkina,b Koen Robeyns,b

    Sjoerd Verkaart,c Joost G. J. Hoenderop,c Mariusz P. Mitoraj,*d Yann Garcia,*b and Debasis Das*a

    aDepartment of Chemistry, The University of Burdwan, 713104, Burdwan, West Bengal, India. Fax: (+91) 342

    2530452; E-mail: [email protected] bInstitute of Condensed Matter and Nanosciences, Molecules, Solids and Reactivity (IMCN/MOST), Université

    Catholique de Louvain, Place L. Pasteur 1, 1348 Louvain-la-Neuve, Belgium. Fax: +32(0) 1047 2330; Tel: +32(0)

    1047 2831; E-mail: [email protected], [email protected] cDepartment of Physiology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center,

    6500 HB Nijmegen, The Netherlands dDepartment of Theoretical Chemistry, Faculty of Chemistry, Jagiellonian University, R. Ingardena 3, 30-060

    Cracow, Poland. E-mail: [email protected]

    10 9 8 7 4 3 2 1

    *

    *

    *

    *

    H, ppm

    l+m+n g+k i hp o

    e+f+g ab+c+d

    *

    *

    *

    *

    Fig. S1 1H NMR spectra of REDA-2PC (black), REDA-2PC + 0.5 equivalent of NCS– (red), REDA-2PC + 1

    equivalent of NCS– (blue), REDA-2PC + 2.5 equivalents of NCS– (purple) and REDA-2PC + 5 equivalents of

    NCS– (wine) in CD3CN:D2O (9:1, v/v). For peak picking in the spectrum of REDA-2PC see Scheme S1. Some

    new bands are marked with an asterisk.

    Electronic Supplementary Material (ESI) for RSC Advances. This journal is © The Royal Society of Chemistry 2015

    mailto:[email protected]

  • Fig. S2 FTIR spectra of REDA-2PC (black) and [REDA-2PC + NCS–] (red)

    Fig. S3 The QTOF mass spectrum of REDA-2PC.

  • 3 4 5 6 7 8 9 10 11 0

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    Fl

    uo re

    sc en

    ce In

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    . u .]

    pH

    Fig. S4 Effect of pH on the emission intensity of REDA-2PC (black) and the [REDA-2PC + NCS−] system (red).

    525 550 575 600 625 650 675 700 0

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    [REDA-2PC + NCS-] [REDA-2PC + ClO4

    -]

    Fl uo

    re sc

    en ce

    In te

    ns ity

    [a . u

    .]

     [nm]

    [REDA-2PC + other ions]

    Fig. S5 Fluorescence spectra of REDA-2PC (50 µM) in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4;

    λexc = 520 nm) solution in the presence of different anions: N3−, NO2−, NO3−, NCO−, AcO−, F−, Cl−, Br−, I−, SO42−,

    C2O42−, Cr2O72−, PO43−, AsO43−, ClO4− and NCS− (500 µM).

  • Fig. S6 UV light exposed colour of REDA-2PC (50 µM) in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH

    7.4) solution in the presence of 500 µM of different anions (from left to right: N3−, NO2−, NO3−, NCO−, AcO−, F−,

    Cl−, Br−, I−, NCS–, SO42−, C2O42−, Cr2O72−, PO43−, AsO43−, ClO4−).

    Fig. S7 UV light exposed colour of REDA-2PC (50 µM) in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH

    7.4) solution upon gradual addition of NCS– (from left to right: 0, 0.001, 0.01, 0.1, 1.0, 3.0, 5.0, 7.0, 10.0, 25.0, 50.0,

    75.0, 100.0, 250.0 and 500.0 µM).

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    Fl uo

    re sc

    en ce

    In te

    ns ity

    [a . u

    .]

    Concentration of NCS- [M]

    Fl uo

    re sc

    en ce

    In te

    ns ity

    [a . u

    .]

    Concentration of NCS- [M]

    Fig. S8 Fluorescence intensity of REDA-2PC (50 µM) vs. externally added NCS– (0.001–500 µM). Inset shows the

    linearity of the fluorescence intensity, observed up to 10 µM of added NCS–.

  • 480 500 520 540 560 580 600 0.0

    0.1

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    0.4

    0.5

    [REDA-2PC + other ions]

    Ab

    so rb

    an ce

     [nm]

    [REDA-2PC + NCS-]

    Fig. S9 Absorption spectra of REDA-2PC (50 µM) in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4)

    solution in presence of different anions: N3−, NO2−, NO3−, NCO−, AcO−, F−, Cl−, Br−, I−, SO42−, C2O42−, Cr2O72−,

    PO43−, AsO43−, ClO4− and NCS− (500 µM).

    Fig. S10 Naked eye view of REDA-2PC (50 µM) in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4)

    solution in presence of 500 µM of different anions (from left to right: only REDA-2PC, N3−, NO2−, NO3−, NCO−,

    AcO−, F−, Cl−, NCS–, Br−, I−, SO42−, C2O42−, Cr2O72−, PO43−, AsO43−, ClO4−).

    Fig. S11 Naked eye view of REDA-2PC (50 µM) HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4) solution

    upon gradual addition of NCS– (from left to right: 0, 0.001, 0.01, 0.1, 1.0, 3.0, 5.0, 7.0, 10.0, 25.0, 50.0, 75.0, 100.0,

    250.0 and 500.0 µM).

  • 0 100 200 300 400 500 0.0

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    A bs

    or ba

    nc e

    Concentration of NCS- [M]

    Ab

    so rb

    an ce

    Concentration of NCS- [M]

    Fig. S12 Variation of absorbance of REDA-2PC (50 µM) at λ = 554 nm in HEPES buffered (0.1 M; CH3CN:H2O,

    9:1 v/v; pH 7.4) solution as a function of externally added NCS– (0.001–500 μM). Inset shows the linearity of the

    absorbance intensity, observed up to 10 µM of added NCS–.

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    Ions

    REDA-2PC [REDA-2PC + NCS-] [REDA-2PC + NCS-] + other ions

    Fig. S13 Emission intensity of the [REDA-2PC (50 µM) + SCN– (500 µM)] system in HEPES buffered (0.1 M;

    CH3CN:H2O, 9:1 v/v; pH 7.4; λexc = 520 nm) solution in presence of different ions (500 µM): N3− (1), NO2− (2),

    NO3− (3), NCO− (4), AcO− (5), F− (6), Cl− (7), Br− (8), I− (9), SO42− (10), C2O42− (11), Cr2O72− (12), PO43− (13),

    AsO43− (14), ClO4− (15), Na+ (16), K+ (17), Mg2+ (18), Ca2+ (19), Fe3+ (20) and a mixture of all ions (21).

  • Fig. S14 Naked eye (bottom) and UV light exposed (top) views of the [REDA-2PC (50 µM) + SCN– (500 µM)]

    system in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4) solution in the presence of different ions (from

    left to right: N3−, NO2−, NO3−, NCO−, AcO−, F−, Cl−, Br−, I−, SO42−, C2O42−, Cr2O72−, PO43−, AsO43−, ClO4−, Na+, K+,

    Mg2+, Ca2+ and Fe3+).

    0 200000 400000 600000 800000 1000000 0

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    (F lim

    - F 0)/

    (F x-

    F 0 )

    Concentration of NCS- [M-1]

    Fig. S15 Determination of the binding constant of REDA with NCS– HEPES buffered (0.1 M; CH3CN:H2O, 9:1

    v/v; pH 7.4; λexc = 520 nm) solution using (Flim – F0)/(Fx – F0) = 1+ (1/K) × (1/[M]n), where Flim, F0 and Fx are

    fluorescence intensities of REDA in the presence of NCS– at saturation, free REDA and at any intermediate NCS–

    concentration, respectively.

  • 0.0 0.2 0.4 0.6 0.8 1.0 120

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    n m

    [a . u

    .]

    [REDA-2PC]/([REDA-2PC] + [NCS-])

    Fig. S16 Job’s plot for the determination of stoichiometry of the [REDA-2PC (50 μM) + NCS–] system in HEPES

    buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4) solution.

    9 8 7 6 5 4 3 H, ppm

    Fig. S17 1H NMR spectrum of FEDA-2PC in CD3CN:D2O (9:1, v/v)

  • Fig. S18 The QTOF mass spectrum of FEDA-2PC.

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    Fig. S19 Fluorescence spectra of FEDA-2PC (50 µM) in HEPES buffered (0.1 M; CH3CN:H2O, 9:1 v/v; pH 7.4;

    λexc = 405 nm) solution in absence (black) and presence (red) of NCS– (500 µM).

  • Fig. S20 The ESI mass spectrum of the species obtained after interaction of REDA-2PC with NCS– in CH3CN:H2O

    (9:1, v/v).

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    RE DA

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    d to

    H 2O

    H2O 0.1 nM NCS - 1 M NCS- 100 M NCS-

    Fig. S21 Effect of the NCS– incubation on the REDA-2PC (10 µM) fluorescence of human embryonic kidney 293

    (HEK293) cells. Error bars represent mean + SEM of at least 90 cells per condition measured at three differe