Facile “Click” to Fabricate a FRET-Based · 2014. 4. 29. · A slurry of 3 (100 mg, 0.20 mmol),...

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Facile “Click” to Fabricate a FRET-Based

Ratiometric Fluorescent Cu2+

Probe

Zhangjun Hu,‡* a, b

, Jiwen Hu,‡a

Yang Cui,a Guannan Wang,

b Xuanjun Zhang,* b

Kajsa Uvdal, b Hong-Wen

Gao a

aState Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai, 200092, P. R.

China

bDivision of Molecular Surface Physics & Nanosciecne, IFM, Linköping University, Linköping, 58183, Sweden

*Corresponding authors. E-mail address: [email protected] (Z. Hu) or [email protected] (X. Zhang)

SYNTHESIS OF RNʹ ....................................................................................................................................................... 2

FIG. S1 ......................................................................................................................................................................... 3

FIG. S2 ......................................................................................................................................................................... 3

FIG. S3 ........................................................................................................................................................................ 4

FIG. S4 ......................................................................................................................................................................... 4

NMR AND MS SPECTRA OF COMPOUNDS ................................................................................................................ 5-14

Electronic Supplementary Material (ESI) for Journal of Materials Chemistry B.This journal is © The Royal Society of Chemistry 2014

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Synthesis of RN′

Scheme S1 Design and synthesis of rhodamine-based fluorescent Cu2+ probes RNʹ.

A slurry of 2-(2-(-2-chloroethoxy)ethoxy)ethanol (840 mg, 5 mmol) and sodium azide (650 mg, 10 mmol) in 20 mL

DMF was stirring at 100oC overnight. After cooling to room temperature, 50 mL dichloromethane was added and the

mixture was washed with brine (3 × 100 mL). The organic phase was dried over anhydrous MgSO4, filtered and

evaporated under vacuum. The crude was purified by flash chromatography using ethyl acetate/heptane (1/1, v/v),

yielding a colourless oil 4 (730 mg, 83 %). 1H NMR (300 MHz, CDCl3) δ (ppm) = 3.72-3.64 (m, 8H), 3.60-3.57 (m,

2H), 3.36 (t, 2 H, J = 5.0), 2.46 (br, 1H); 13C NMR (75 MHz, CDCl3) δ (ppm) = 72.71, 70.86, 70.60, 70.23, 61.94,

50.87.

A slurry of 3 (100 mg, 0.20 mmol), 4 (35 mg, 0.20 mmol), CuI (3.8 mg, 0.02 mmol ) and N,N-diisopropylethylamine

(26 mg, 0.20 mmol) in 5 mL THF was stirring at room temperature overnight. The reaction mixture was quenched

with 5 mL brine, and extracted with EtOAc (3 × 10 mL). The organic phase was dried over anhydrous Na2SO4, filtered

and evaporated under vacuum. The crude was by flash column chromatography ethyl acetate/methanol (10/1, v/v),

yielding a whitish foamy solid RNʹ (123 mg, 91%). 1H NMR (300 MHz, CDCl3) δ (ppm) = 7.95–7.86 (m, 1H), 7.79

(s, 1H), 7.51–7.39 (m, 2H), 7.13 – 7.03 (m, 1H), 6.42 (dd, J = 8.7, 5.7 Hz, 4H), 6.24 (dd, J = 8.8, 2.6 Hz, 2H), 4.44 (t, J

= 5.0 Hz, 2H), 3.90 (d, J = 4.9 Hz, 2H), 3.86 – 3.78 (m, 2H), 3.73 (s, 2H), 3.66 – 3.50 (m, 6H), 3.33 (q, J = 7.0 Hz,

8H), 1.16 (t, J = 7.1 Hz, 12H); 13C NMR (75 MHz, CDCl3) δ (ppm) = 167.15, 153.85, 152.05, 149.00, 145.07, 132.94,

129.99, 128.63, 128.24, 124.11, 124.00, 123.02, 108.06, 105.48, 98.14, 72.71, 70.61, 70.38, 69.55, 66.00, 61.76,

50.11, 46.91, 44.48, 12.79; MS (ESI): m/z (%): 670.2 (100) M++H. Anal. Calcd for C37H47N7O5 (%): C, 66.35; H,

7.07; N, 14.64. Found: C, 66.53; H, 7.03; N, 14.57.

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Supporting Figures

450 500 550 600 6500.00

0.05

0.10

0.15

0.20

Abs

orba

nce

Wavelength (nm)

RN' and RN'+ other ions

RN'+ Cu2+ '

540 560 580 600 620 6400

200

400

600

800

1000

Flu

ores

cenc

e In

tens

ity (

a.u.

)

Wavelength (nm)

RN' + Cu2+

RN' and RN' + other ions

Fig. S1 UV/vis and fluorescence spectra of RN′ (5 µM) in the presence of various metal ions (25 µM): Na+, K

+, Ca

2+, Mg

2+, Al

3+,

Cr3+

, Pb2+

, Fe2+

, Fe3+

, Co2+

, Ni2+

, Ba2+

, Hg2+

, Mn2+

, La3+

, Zn2+

, Cd2+

, Cu+ and Cu

2+ in CH3CN/H2O (1:1, v/v) buffered with hepes (pH

7.4, 20 mM).

0 20 40 60 80 100 120

0

4

8

12

16

[Cu2+]/µµµµm

10 20 30 40 500

2

4

6

8

10

[Cu2+]/µµµµm

I 568/I 54

0I 568

/I 540

Fig. S2 The ratio (I568/I540) of RN (5 μM) upon addition of Cu2+

(0-120 μM), in CH3CN/H2O (1:1, v/v) buffered with hepes (pH 7.4,

20 mM). Inset: fitting curves of the ratio (I568/I540) of RN (5 μM) upon addition of Cu2+

in 10-50 μM.

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0 100000 200000 300000 400000

0

2

4

6

8

Y=A+BXA=0.4205B=2.031E-5R=0.9929

R0/

(R-R

0)

1/[Cu 2+]

Fig. S3 Fitting Curve of fluorescence intensity ratio R0/(R-R0) at 568/540 of RN upon 1/[Cu2+

], Ka = A/B =2.07 × 104 M

-1

The binding constant was calculated from the fluorescence intensity-titration curve according to the equation: 1

1][M]a)][1/K[a/(b)R/(RR a00 +−=−

Where R0 is the emission intensity ratio (I568/I540) of RN, R is the emission intensity ratio (I568/I540) of RN upon addition of

different amounts of Cu2+

. [M] is the concentration of Cu2+

. a and b are constants. The association constant values Ka is

calculated by the ratio intercept/slope.

300 350 400 450 500 550 600 650 7000.0

0.2

0.4

0.6

0.8

1.0

1.2

Abs

orba

nce

Wavelength (nm)

RN2 RN2 + Cu2+

Fig. S4 UV absorption spectra of RN (10 μM) and RN (10 μM) in the presence of Cu2+

(0.2 mM) in CH3CN/H2O (1:1, v/v)

buffered with hepes (pH 7.4, 20 mM).

1 Forgues, F. S., Le Bris, M. T., Guett, J. P., Valeur, B., J. Phys. Chem.1988, 92, 6233.

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NMR and MS spectra of compounds

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SQ__descrip #637 RT: 5.60 AV: 1 NL: 1.07E8T: {0;0} + c ESI corona sid=50.00 det=1600.00 Full ms [ 75.00-2000.00]

100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950m/z

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Rel

ativ

e A

bund

anc

e

495.2

496.2

497.3

493.2 596.4565.3509.0 597.4

456.2426.1 669.4


100 200 300 400 500 600 700 800 900 1000m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Rel

ativ

e A

bund

anc

e

670.2

671.3

672.3

655.2

673.4802.5772.3626.6 700.3 967.2343.3 534.4457.3 877.9 896.2508.4 1023.3425.8 837.8141.682.6 174.6 195.2 228.8 274.1 372.1 599.0

O

N

N N

O

NH N

NN

Exact Mass: 669.36

OO

OH

O

N

N N

O

NH

Exact Mass: 494.27

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100 150 200 250 300 350 400 450 500 550 600m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100R

ela

tive

Ab

und

an

ce327.0

328.9

330.0 367.9326.1 331.0 370.0

234.0 260.9217.9 343.2157.2 465.9290.8 405.6 493.2 508.291.3 132.8 619.1552.0 573.4


100 150 200 250 300 350 400 450 500 550 600 650m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Rel

ativ

e A

bun

dan

ce

334.0

335.2

375.0336.1

376.2337.2277.0 305.9264.9 397.0 667.2471.6227.8173.9 485.5155.0 213.3109.996.3 437.5 507.6 615.2595.8542.2 556.2 655.7

N

S

NCl

OO

Mol. Wt.: 326.84

N

S

NN3

OO

Mol. Wt.: 333.41

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SQ_657 #738 RT: 6.49 AV: 1 NL: 1.13E8T: {0;1} + c ESI corona sid=75.00 det=1600.00 Full ms [ 75.00-2000.00]

200 400 600 800 1000 1200 1400 1600 1800 2000m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Re

lativ

e A

bun

da

nce

828.1

830.1

826.1 1657.0832.3238.4 660.7297.3205.3 405.9 930.0565.9443.8 1094.1 1605.51411.8 1723.91242.2 1805.9987.9 1985.31342.2

Facile “Click” to Fabricate a FRET-Based · 2014. 4. 29. · A slurry of 3 (100 mg, 0.20 mmol),...

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