Supplementary Information
A Fluorescent Molecular Ruler as a Selective Probe for ω-Aminoacids
Daniel Moreno, José V. Cuevas, Gabriel García-Herbosa, and Tomás Torroba*
Department of Chemistry, Faculty of Science, University of Burgos, 09001 Burgos, Spain.
Experimental Part:
Materials and methods: The reactions performed with air sensitive reagents were conducted under dry
nitrogen. The solvents were previously distilled under nitrogen over calcium hydride or sodium filaments.
Melting points were determined in a Gallenkamp apparatus and are not corrected. Infrared spectra were
registered in a Nicolet Impact 410 spectrometer in potassium bromide tablets. NMR spectra were
recorded in Varian Mercury-300 and Varian Unity Inova-400 machines, in DMSO-d6, CDCl3, CD3CN,
CD3OD. Chemical shifts are reported in ppm with respect to residual solvent protons, coupling constants
(JX-X’) are reported in Hz. Elemental analyses of C, H and N were taken in a Leco CHNS-932. Mass
spectra were taken in a Micromass AutoSpec machine, by electronic impact at 70 eV. Quantitative UV-
visible measures were performed with a Varian, Cary 300 Bio UV spectrophotometer, in 1 cm UV cells at
25ºC. Fluorescence spectra were recorded in a Varian Cary Eclipse spectrofluorometer, in 1 cm quarz
cells at 25ºC.
Synthesis of 5-(4-aminophenyl)indan-1-one
Br
O
Pd(PPh3)4/CsCO3
O
H2N
3
+
B
NH2
O O
21
Pd(PPh3)4 (54 mg, 0.047 mmol) was added under nitrogen to a solution of 5-bromo-1-indanone 1 (100 mg
0.470 mmol) in toluene/butanol 4:1 mixture (15 mL) and the mixture was stirred for 30 minutes at room
temperature. Then solid 4-aminobenzeneboronic pinacol ester 2 (103 mg, 0.470 mmol) and cesium
carbonate (463 mg, 1.42 mmol) dissolved in H2O (2 mL) were added and the mixture was heated under
reflux for 16 h. Then the solvent was evaporated under reduced pressure and the residue was added to
water (20 mL) and extracted with CH2Cl2 (3 x 30 mL). The combined organic extracts were dried
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(Na2SO4) and evaporated and the residue was purified by flash chromatography (silica, 3 x 30 cm),
CH2Cl2, to get 5-(4-aminophenyl)indan-1-one 3 as a light yellow solid (95 mg, 91 %), mp 250-251 ºC.
IR (KBr, cm-1): 3429 and 3335 (NH2), 1678 and 1592 (C=O), 1297 and 1278, 811. 1H NMR (DMSO-d6, 400MHz) 7.71 (s, 1H, ArH), 7.60 (s, 2H, ArH), 7.48 (d, J = 6 Hz, 2H, ArH), 6.68 (d,
J = 6 Hz, 2H, ArH), 5.44 (s, br, exch, 2H, NH2), 3.10 (m, 2H, CH2), 2.61 (m, 2H, CH2). 13C NMR (DMSO-d6, 100MHz) : 205.44 (C=O), 156.14 (C-NH2), 149.53, 146.91, 134.05, 127.90 (CH),
125.84, 124.60 (CH), 123.25 (CH), 122.79 (CH), 114.13 (CH), 36.10 (CH2), 25.44 (CH2).
MS (EI) m/z (%): 224 (M++1, 17), 223 (M+, 100), 195 (18), 167 (8), 152 (9), 139 (4), 117 (7), 96 (3).
HRMS (EI): calcd. for C15H13NO: 223.0997 (M+); found: 223.0997.
Anal. Calcd for C15H13NO: C, 80.69; H, 5.87; N, 6.27. Found: C: 80.44; H: 5.95; N: 6.11.
1H-NMR spectrum (DMSO-d6, 400 MHz) of 5-(4-aminophenyl)indan-1-one
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30405060708090100110120130140150160170180190200210f1 (ppm)
25
.44
36
.10
11
4.1
3
12
2.7
91
23
.25
12
4.6
01
25
.84
12
7.9
01
34
.05
14
6.9
11
49
.53
15
6.1
4
20
5.4
4
13C-NMR spectrum (DMSO-d6, 100 MHz) of 5-(4-aminophenyl)indan-1-one
DEPT-NMR spectrum (DMSO-d6, 100 MHz) of 5-(4-aminophenyl)indan-1-one
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Synthesis of 1-dicyanomethylene-5-(4-aminophenyl)indane 4
Malononitrile (50 mg 0.76 mmol) and DABCO (30 mg, 0.23 mmol) were added under nitrogen to 5-(4-
aminophenyl)indan-1-one 3 (50 mg, 0.23 mmol) dissolved in toluene (10 mL). The mixture was heated
under reflux for 2 h and then added to water (30 mL), stirred for 5 minutes and extracted with
dichloromethane (5 x 20 mL). The combined organic extracts were dried (Na2SO4) and the solvent
evaporated under reduced pressure. The crude product was purified by flash chromatography (silica, 3 x
30 cm), CH2Cl2, to get 1-dicyanomethylene-5-(4-aminophenyl)indene 4 as an orange solid (59 mg, 95 %),
mp 320-322 ºC (decomp).
IR (KBr, cm-1): 3491 and 3382 (NH2), 2221 (CN), 1627, 1600 and 1561 (C=C). 1H NMR (DMSO-d6, 300MHz): 8.14 (d, J = 9 Hz, 1H, ArH), 7.75 (m, 2H, ArH), 7.54 (d, J = 9 Hz, 2H,
ArH), 6.66 (d, J = 9 Hz, 2H, ArH), 5.62 (s, br, exch, 2H, NH2), 3.23 (m, 2H, CH2), 3.23 (m, 2H, CH2). 13C NMR (DMSO-d6, 75 MHz): 179.83, 156.43, 150.35, 147.42, 132.53, 128.20 (CH), 125.34 (CH),
124.90 (CH), 124.72, 121.69 (CH), 114.36 (CN), 114.15 (CH), 113.95 (CN), 69.96 (C(CN)2), 34.66
(CH2), 29.41 (CH2).
MS (EI) m/z (%): 272 (M++1, 20), 271(M+, 100), 243 (3), 214 (2), 206 (1), 204 (2), 189 (2), 135 (2).
HRMS (EI): calcd. for C18H13N3: 271.1109 (M+); found: 271.1108
Anal. Calcd for C18H13N3: C, 79.68; H, 4.83; N, 15.49. Found: C: 79.77; H: 4.97; N: 15.35.
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1H NMR spectrum (DMSO-d6, 300 MHz) of 1-dicyanomethylene-5-(4-aminophenyl)indane
13C NMR spectrum (DMSO-d6, 75 MHz) of 1-dicyanomethylene-5-(4-aminophenyl)indane
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Expansion of the 13C NMR spectrum (DMSO-d6, 75 MHz) of 1-dicyanomethylene-5-(4-
aminophenyl)indane
DEPT-NMR spectrum (DMSO-d6, 75 MHz) of 1-dicyanomethylene-5-(4-aminophenyl)indane
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Expansion of the DEPT-NMR spectrum (DMSO-d6, 75 MHz) of 1-dicyanomethylene-5-(4-
aminophenyl)indane
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Synthesis of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6
1-Dicyanomethylene-5-(4-aminophenyl)indane 4 (100 mg, 0.38 mmol) was added to m-xylylene
diisocyanate 5 (15 µL, 0.095 mmol) dissolved in CHCl3 (20 mL) and the mixture was stirred at room
temperature for 24 h. Then a second portion of m-xylylene diisocyanate (15 µL, 0.095 mmol) was added
and the mixture was stirred for additional 24 h. The solid formed after this time was filtered off, washed
with CHCl3 (2 portions of 2 mL) and dried under reduced pressure. Compound 6 was obtained as a
yellow solid (104 mg, 75%), mp 212-214ºC.
IR (KBr, cm-1): 3328 (br, NH), 2221(C≡N), 1658, 1604 and 1557 (C=O), 1320, 1219, 1184. 1H NMR (DMSO-d6 , 400MHz) 8.82 (s, 2H, NH) 8.17 (d, J = 6 Hz, 2H, ArH), 7.82 (s, 2H, ArH), 7.79 (d,
J = 6 Hz, 2H, ArH), 7.65 (d, J = 6 Hz, 4H, ArH), 7.52 (d, J = 6 Hz, 4H, ArH), 7.29 (m, 2H, ArH), 7.20
(m, 2H, ArH), 6.71 (t, J = 6 Hz, 2H, NH), 4.32 (d, J = 6 Hz, 4H, CH2) , 3.28 (m, 4H, CH2), 3.15 (m, 4H,
CH2). 13C NMR (DMSO-d6 , 100MHz) 179.60, 155.78, 154.63, 146.04, 141.21, 140.00, 133.07, 130.10, 128.22
(CH), 127.58 (CH), 125.72, 125.67 (CH), 125.55 (CH), 125.22 (CH), 122.60 (CH), 117.54 (CH), 113.66
(CN), 113.25 (CN), 70.61, 42.32 (CH2), 34.24 (CH2), 29.04 (CH2).
MS (FAB) m/z (+): 731.40 (M++1). ESI MS m/z (%):753.28 (M+Na+), 731.29 (M++1)
Anal. Calcd for C46H34N8O2: C, 75.60; H, 4.69; N, 15.33. Found: C, 75.42; H, 4.84; N, 15.09.
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1H NMR spectrum (DMSO-d6, 400 MHz) of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene
13C NMR spectrum (DMSO-d6, 100 MHz) of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene
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DEPT-NMR spectrum (DMSO-d6, 100 MHz) of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene
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Synthesis of 1-benzyl-3-[4-(1-(dicyanomethyleneindan-5-yl)phenyl]urea 8:
1-Dicyanomethylene-5-(4-aminophenyl)indane 4 (50 mg, 0.18 mmol) was added to benzyl isocyanate 7
(25 mg, 0.18 mmol) dissolved in CHCl3 (10mL) and the mixture was stirred at room temperature for 48 h.
Hexane (2 mL) was then added and the solid formed was filtered off, washed with CHCl3 (2 portions of 2
mL) and dried. Compound 8 was obtained as a yellow solid (56 mg, 75%), mp 189-190 ºC.
IR (KBr, cm-1) 3336 (br, NH), 2221 (C≡N), 1639 and 1565 (C=O), 1328, 1227. 1H NMR (DMSO-d6 , 400MHz): 8.74 (s, 1H, NH) 8.25 (d, J = 8 Hz, 1H, ArH), 7.87 (s, 1H, ArH), 7.83 (d,
J = 8 Hz, 1H, ArH), 7.71 (d, J = 8 Hz, 2H, ArH), 7.56 (d, J = 8 Hz, 2H, ArH), 7.34-7.32 (m, 4H, ArH),
7.25 (m, 1H, ArH), 6.66 (t, J = 6 Hz, 1H, NH), 4.33 (d, J = 6 Hz, 2H, CH2), 3.30 (m, 2H, CH2), 3.19 (m,
2H, CH2). 13C NMR (DMSO-d6, 100MHz) 179.68, 155.96, 154.82, 146.36, 141.44, 139.96, 133.40, 130.55, 128.08,
127.44, 126.96, 126.57, 125.63, 125.17, 122.87 (CHAr), 117.85, 113.81 (CN), 113.40 (CN), 71.00, 42.69
(CH2), 34.50 (CH2), 29.28 (CH2).
MS (FAB) m/z (+): 405.06 (M++1). MS (EI) m/z (%): 404 (M+, 10), 297 (95), 271 (100).
HRMS (EI): calcd. for C26H20N4O: 404.1637 (M+); found: 404.1633.
Anal. Calcd for C26H20N4O: C, 77.21; H, 4.98; N, 13.85. Found: C, 77.06; H, 5.12; N, 13.67.
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1H NMR spectrum (DMSO-d6, 400 MHz) of 1-benzyl-3-[4-(1-(dicyanomethyleneindan-5-yl)phenyl]urea
13C NMR spectrum (DMSO-d6, 100 MHz) of 1-benzyl-3-[4-(1-(dicyanomethyleneindan-5-yl)phenyl]urea
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112113114115116117118119121123125127129131133135f1 (ppm)
An expansion of the 13C NMR spectrum (DMSO-d6, 100 MHz) of 1-benzyl-3-[4-(1-(dicyanomethyleneindan-
5-yl)phenyl]urea
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Solvatochromism of 1-dicyanomethylene-5-(4-aminophenyl)indane 4:
(1) (2) (3) (4) (5) (6) (7) 10-4 M solutions of 4 in (1) Hexane, (2) Et2O, (3) THF, (4) CH2Cl2, (5) MeCN, (6) MeOH, (7) DMSO
0
0.2
0.4
0.6
0.8
1
350 400 450 500 550 600wavelength (nm)
no
rmal
ized
ab
sorb
ance
HEXANE
ETHER
THF
DICHLOROMETANE
ACETONITRILE
METHANOL
DMSO
Normalized absorbance spectra of 10-4 M solutions of 4 in Hexane, Et2O, THF, CH2Cl2, MeCN, MeOH, DMSO
H2N
NC
CN
4 (1) (2) (3) (4) (5) (6) (7) λexc = 366 nm 10-4 M solutions of 4 in (1) Hexane, (2) Et2O, (3) THF, (4) CH2Cl2, (5) MeCN, (6) MeOH, (7) DMSO
0
0.2
0.4
0.6
0.8
1
375 425 475 525 575 625 675Wavelength (nm)
I/Io
HEXANE
ETHER
THF
DICHLOROMETHANE
ACETONITRILE
METHANOL
DMSO
Normalized fluorescente spectra of 10-4 M solutions of 4 in Hexane, Et2O, THF, CH2Cl2, MeCN,
MeOH, DMSO; λexc = 366 nm.
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Behavior of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6 in MeCN and
DMSO in the presence of common anions:
Ref. F- Cl- Br- I- BzO- NO3- H2PO4
- HSO4- AcO- CN- SCN-
10-4 M Solutions of 6 in MeCN +2 equivalents of anion: F-, Cl-, Br-, I-, BzO-, NO3-, H2PO4
-, HSO4-, AcO-,
CN-, SCN-
Ref. F- Cl- Br- I- BzO- NO3- H2PO4
- HSO4- AcO- CN- SCN-
10-4 M Solutions of 6 in MeCN +2 equivalents of anion: F-, Cl-, Br-, I-, BzO-, NO3-, H2PO4
-, HSO4-, AcO-,
CN-, SCN-; λexc = 366nm
Ref. F- Cl- Br- I- BzO- NO3- H2PO4
- HSO4- AcO- CN- SCN-
10-4 M Solutions of 6 in DMSO +2 equivalents of anion: F-, Cl-, Br-, I-, BzO-, NO3-, H2PO4
-, HSO4-, AcO-,
CN-, SCN-
Ref. F- Cl- Br- I- BzO- NO3- H2PO4
- HSO4- AcO- CN- SCN-
10-4 M Solutions of 6 in DMSO +2 equivalents of anion: F-, Cl-, Br-, I-, BzO-, NO3-, H2PO4
-, HSO4-, AcO-,
CN-, SCN-; λexc = 366 nm
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Fluorimetric titrations of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6
in DMSO with common anions:
F-DMSO
0
10
20
30
40
50
60
70
80
400 450 500 550 600 650Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.00000 0.00005 0.00010 0.00015 0.00020 0.0002510
20
30
40
50
60
70
80
complex 1:1 Chi^2 = 3.81884R^2 = 0.99156 Ao 78.32304 ±1.15304Af 11.15866 ±2.668Co 0.00009 ±5.5948E-6K 146856.21076 ±64388.08958
Inte
nsi
ty
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with F- (excitation = 290 nm).
Right: Titration profile fitted to a 1:1 model (λmax = 515 nm) for the titration of 6 with F-
BzO- DMSO
0
20
40
60
80
100
120
140
450 500 550 600 650 700Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.00040
20
40
60
80
100
complejo 1:1 Quitando los 2 primeros puntos Chi^2 = 0.0917R^2 = 0.99977 Ao 158.11626 ±3.15139Af 7.99681 ±0.16131Co 0.00001 ±8.0069E-7K 131190.53066 ±1787.80021
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with BzO- (excitation = 396
nm). Right: Titration profile fitted to a 1:1 model (λmax = 555 nm) for the titration of 6 with BzO-
H2PO4- DMSO
0
10
20
30
40
50
60
70
80
450 500 550 600Wavelength (nm)
Inte
ns
ity
(a
.u.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.000510
20
30
40
50
60
70
80complejo 1:1 DEJANDO c=CTE
Chi^2 = 1.99719R^2 = 0.99453 Ao 78.39944 ±0.56336Af 17.3972 ±0.68597Co 0.0001 ±0K 92906.65808 ±10696.21957
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with H2PO4- (exc = 290 nm).
Right: Titration profile fitted to a 1:1 model (λmax = 515 nm) for the titration of 6 with H2PO4-
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AcO- DMSO
0
20
40
60
80
100
120
140
400 450 500 550 600 650Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005
40
60
80
100
120
140
160
complex 1:1 Chi^2 = 14.31229R^2 = 0.98801 Ao 160.10738 ±1.47945Af 40 ±0Co 0.0001 ±0K 31868.16814 ±1766.98407
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with AcO- (exc = 290 nm).
Right: Titration profile fitted to a 1:1 model (λmax = 515 nm) for the titration of 6 with AcO-
CN- DMSO
0
10
20
30
40
50
60
70
80
420 470 520 570 620Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005
10
20
30
40
50
60
70
80
90
complex 1:1
Chi^2 = 4.68425R^2 = 0.99192 Ao 89.28482 ±1.48214Af 14.8314 ±0.98686Co 0.00008 ±2.6546E-6K 1051857.84153±535334.04094
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with CN- (exc = 290 nm).
Right: Titration profile fitted to a 1:1 model (λmax = 515 nm) for the titration of 6 with CN-
BzO- DMSO:H2O(9:1)
0
10
20
30
40
50
60
450 500 550 600 650Wavelength (nm)
Inte
ns
ity
0.0000 0.0001 0.0002 0.0003 0.0004 0.000510
20
30
40
50
60
complex 1:1 Chi^2 = 3.83587R^2 = 0.97474 Ao 54.88309 ±0.77198Af 17.22144 ±0.91885Co 0.0001 ±0K 167026.89433 ±54717.90414
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO:H2O (9:1)) with BzO- (exc =
396 nm). Right: Titration profile fitted to a 1:1 model (λmax = 555 nm) for the titration of 6 with BzO-
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AcO- DMSO:H2O(9:1)
0
10
20
30
40
50
60
450 500 550 600 650 700Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.00050
10
20
30
40
50
60complex 1:1 Chi^2 = 0.59147R^2 = 0.99799 Ao 61.56569 ±0.41839Af 5.96347 ±0.57055Co 0.0001 ±4.2261E-6K 57097.03899 ±6778.11165
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO:H2O (9:1)) with AcO- (exc =
396 nm). Right: Titration profile fitted to a 1:1 model (λmax = 555 nm) for the titration of 6 with AcO-
CN- DMSO:H2O(9:1)
0
50
100
150
200
250
450 500 550 600 650 700Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.00050
50
100
150
200
250complex 1:1 Chi^2 = 26.64141R^2 = 0.99379 Ao 231.84437 ±2.09644Af 32.00637 ±1.74664Co 0.00009 ±0K 480213.59697 ±105297.44565
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO:H2O (9:1)) with CN- (exc =
396 nm). Right: Titration profile fitted to a 1:1 model (λmax = 555 nm) for the titration of 6 with CN-
F- DMSO:H2O (9:1)
0
10
20
30
40
50
450 500 550 600 650Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005
15
20
25
30
35
40
45
50
55complex 1:1 Chi^2 = 0.66835R^2 = 0.99426 Ao 50.77066 ±0.32986Af 16.90096 ±0.44674Co 0.0001 ±0K 73740.60155 ±8926.16566
Inte
nsi
ty
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO:H2O (9:1)) with F- (exc = 396
nm). Right: Titration profile fitted to a 1:1 model (λmax= 555 nm) for the titration of 6 with F-
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Job's plot analysis of fluorescence titrations of 6 with F-
Job's plot analysis of fluorescence titrations of 6 with F- revealed a maximum at a 50% mole fraction, in
accord with the proposed 1:1 binding stoichiometry
-5
0
5
10
15
20
0 0.2 0.4 0.6 0.8 1
XF-
XF-x
Inte
ns
ity
Job's plot analysis of fluorescence titrations of 6 with F- in MeCN (exc = 396 nm) (em = 557 nm)
Complexation constants for compound 6 and common anions
Anions Complex logK1 R2
BzO- 1:1 5.118 ± 0.006 0.999 AcO- 1:1 4.503 ± 0.023 0.988 CN- 1:1 6.022 ± 0.179 0.992 F- 1:1 5.169 ± 0.158 0.991
H2PO4- 1:1 4.968 ± 0.047 0.994
Complexation constants for compound 6 in DMSO and common anions
Anions Complex logK1 R2
BzO- 1:1 5.118 ± 0.006 0.999 AcO- 1:1 4.503 ± 0.023 0.988 CN- 1:1 6.022 ± 0.179 0.992 F- 1:1 5.169 ± 0.158 0.991
H2PO4- 1:1 4.968 ± 0.047 0.994
Complexation constants for compound 6 in DMSO:H2O (9:1) and common anions
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Fluorimetric titrations of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6
in DMSO with α-aminoacids:
0
10
20
30
40
50
60
420 470 520 570 620
Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005 0.0006 0.0007
30
35
40
45
50
55
60
65
complejo1:1 Chi 2 = 0.52045R 2 = 0.99418 Ao 63.33815 ±0.68132Af 24.58799 ±1.12648Co 0.0001 ±0K 9478.00549 ±1199.41294
Inte
nsity
(a.
u.)
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with L-arginine in water (exc
= 306 nm). Right: Titration profile fitted to a 1:1 model (λmax = 511 nm) for the titration of 6 with L-
arginine
0
20
40
60
80
100
120
140
160
180
420 470 520 570 620Wavelength (nm)
Inte
ns
ity
(a
.u.)
0.0000 0.0002 0.0004 0.0006 0.0008
80
100
120
140
160
180complejo1:1 Chi 2 = 5.24383R^2 = 0.9946 Ao 175.46372 ±1.5268Af 62.09588 ±2.63464Co 0.0001 ±0K 8535.51692 ±844.18975
Inte
nsity
(a.
u.)
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with L-asparagine in water
(exc = 306 nm). Right: Titration profile fitted to a 1:1 model (λmax = 511 nm) for the titration of 6 with L-
asparagine.
0 equiv
6 equiv
0 equiv
8 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
0
10
20
30
40
50
60
70
80
90
400 450 500 550 600 650 700Wavelength (nm)
Inte
ns
ity
(a
.u.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005
30
40
50
60
70
80
90
complejo1:1 Chi 2 = 0.96289R 2 = 0.99598 Ao 80.40854 ±0.68865Af 25.94513 ±1.21473Co 0.0001 ±0K 14641.88662 ±1544.09989
Inte
nsi
ty (
a.u
.)
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with L-glutamine in water
(exc = 306 nm). Right: Titration profile fitted to a 1:1 model (λmax = 511 nm) for the titration of 6 with L-
glutamine.
0
10
20
30
40
50
60
70
425 475 525 575 625 675Wavelength (nm)
Inte
ns
ity
(a
.u.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005 0.0006 0.0007
35
40
45
50
55
60
65
70 complejo1:1 Chi 2 = 0.37202R 2 = 0.99663 Ao 69.06404 ±0.40442Af 23.51426 ±1.66134Co 0.0001 ±0K 4602.27513 ±477.19618
Inte
nsity
(a.
u.)
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with L-lysine in water (exc =
306nm). Right: Titration profile fitted to a 1:1 model (λmax = 511 nm) for the titration of 6 with L-lysine
0 equiv
5 equiv
0 equiv
6 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Job's plot analysis of fluorescence titrations of 6 with L-lysine and L-asparagine
Job's plot analysis of fluorescence titrations of 6 with L-lysine and L-asparagine revealed maximum peaks
at 50% mole fraction, in accord with the proposed 1:1 binding stoichiometry
0
10
20
30
40
50
0 0.2 0.4 0.6 0.8 1Xlysine
Xly
sin
exIn
ten
sity
0
5
10
15
20
25
0 0.2 0.4 0.6 0.8 1Xasparagine
Xa
sp
ara
gin
exIn
ten
sity
Left: Job's plot analysis of fluorescence titrations of 6 (10-4 M in DMSO) with L-lysine in water (exc =
350 nm) (em = 515 nm). Right: Job's plot analysis of fluorescence titrations of 6 (10-4 M in DMSO) with
L-asparagine in water (exc = 396 nm) (em = 557 nm)
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Fluorimetric titrations of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6
in DMSO with ω-aminoacids:
0
10
20
30
40
50
60
70
80
450 500 550 600 650 700Wavelength (nm)
Inte
nsit
y (a
.u.)
0.0000 0.0002 0.0004 0.0006 0.00080.0
0.2
0.4
0.6
0.8
1.0 Model: complejo 2:1 Chi 2 = 0.00006R^2 = 0.99888 Co 0.0001 ±0K1 30661.1342 ±1331.09082K2 8433.74738 ±1872.32341Ao 1.01019 ±0.00672Af 0.16621 ±0.00992b -160.76936 ±166.02104
I/Io
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with β-alanine in water (exc=
396 nm). Right: Titration profile fitted to a 2:1 model (λmax = 562 nm) for the titration of 6 with β-alanine.
0
50
100
150
200
250
450 500 550 600 650 700
Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0002 0.0004 0.0006 0.00080.0
0.2
0.4
0.6
0.8
1.0
Model: complex 2:1 Chi^2 = 0.00004R^2 = 0.99933 Co 0.0001 ±0K1 14046.16698 ±644.88932K2 12683.49195 ±2084.75033Ao 1.00998 ±0.00556Af 0.13514 ±0.00854b -180.80111 ±436.82904I/
Io
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with γ-aminobutiric acid in
water (exc = 396 nm). Right: Titration profile fitted to a 2:1 model (λmax = 562 nm) for the titration of 6
with γ-aminobutiric acid.
0 equiv
8 equiv
0 equiv
8 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
0
20
40
60
80
100
120
140
450 500 550 600 650 700
Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0002 0.0004 0.0006 0.00080.0
0.2
0.4
0.6
0.8
1.0
Model: complejo 2:1 Chi^2 = 0.00002R^2 = 0.99973 Co 0.0001 ±0K1 9596.28128 ±815.54057K2 147933.57307 ±16545.99678Ao 1.00253 ±0.00381Af 0.09748 ±0.00202b -429.44422 ±714.22269I/
Io
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with 5-aminovaleric acid in
water (exc = 396 nm). Right: Titration profile fitted to a 2:1 model (λmax = 562 nm) for the titration of 6
with 5-aminovaleric acid.
0
50
100
150
200
450 500 550 600 650 700Wavelength (nm)
Inte
ns
ity
(a.u
.)
0.0000 0.0001 0.0002 0.0003 0.0004 0.0005 0.0006 0.00070.0
0.2
0.4
0.6
0.8
1.0
Model: complejo 2:1 Chi^2 = 0.00001R^2 = 0.9998 Co 0.0001 ±0K1 4963.08488 ±792.53822K2 378157.69636 ±67242.39871Ao 1.00021 ±0.00338Af 0.09165 ±0.00192b -2665.67851 ±1499.6116I/
Io
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with 6-aminohexanoic acid in
water (exc = 396 nm). Right: Titration profile fitted to a 2:1 model (λmax= 562 nm) for the titration of 6
with 6-aminohexanoic acid.
0 equiv
8 equiv
0 equiv
6 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
0
10
20
30
40
50
60
70
80
90
100
450 500 550 600 650 700Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0002 0.0004 0.0006 0.00080.0
0.2
0.4
0.6
0.8
1.0 Model: complejo 2:1 Chi^2 = 0.00002R^2 = 0.99955 Co 0.0001 ±0K1 15193.02869 ±1825.44451K2 321035.15614 ±49096.59229Ao 0.99983 ±0.00442Af 0.08496 ±0.00168b 986.08083 ±949.92876
I/Io
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with 7-aminoheptanoic acid in
water (exc = 396 nm). Right: Titration profile fitted to a 2:1 model (λmax = 562 nm) for the titration of 6
with 7-aminoheptanoic acid.
Job's plot analysis of fluorescence titrations of 6 with 6-aminohexanoic acid
Job's plot analysis of fluorescence titrations of 6 with 6-aminohexanoic acid revealed a maximum at 66%
mole fraction, in accord with the proposed 2:1 binding stoichiometry
0
10
20
30
40
50
60
70
0 0.2 0.4 0.6 0.8 1X5-aminohexanoic
X5-
amin
oh
exan
oicxI
nte
nsi
ty
Job's plot analysis of fluorescence titrations of 6 (10-4 M in DMSO) with 6-aminohexanoic acid in water
(exc = 350 nm) (em = 515 nm).
0 equiv
8 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Fluorimetric titrations of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6
in DMSO with gabapentin and pregabalin in water:
0
10
20
30
40
50
60
70
80
450 500 550 600 650 700
Wavelength (nm)
Inte
ns
ity
(a
.u.)
0.0000 0.0005 0.0010 0.0015 0.0020 0.002530
40
50
60
70
80 complejo 2:1 Chi^2 = 1.03872R^2 = 0.99369 Co 0.0001 ±0K1 8048.57001 ±7489.43154K2 1028.00328 ±830.97575Ao 79.23813 ±0Af 26.20715 ±3.83709b 593356.59337 ±152143.42604
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with gabapentin in water (exc
= 428 nm). Right: Titration profile fitted to a 2:1 model (λmax = 555 nm) for the titration of 6 with
gabapentin.
0
10
20
30
40
50
60
70
80
450 500 550 600 650 700Wavelength (nm)
Inte
ns
ity
(a
.u.)
0.0000 0.0003 0.0006 0.0009 0.0012 0.0015 0.0018
10
20
30
40
50
60
70
80complex 2:1 Chi^2 = 0.66205R^2 = 0.99817 Co 0.0001 ±0K1 799.52569 ±49.47501K2 1139.54936 ±246.55148Ao 76.1475 ±0Af 11.3929 ±0b -323927.50203 ±28491.10108
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with pregabalin in water (exc =
426 nm). Right: Titration profile fitted to a 2:1 model (λmax= 554 nm) for the titration of 6 with
pregabalin.
0 equiv
20 equiv
0 equiv
18 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Job's plot analysis of fluorescence titrations of 6 with gabapentin
Job's plot analysis of fluorescence titrations of 6 with gabapentin acid revealed a maximum at 66% mole
fraction, in accord with the proposed 2:1 binding stoichiometry
0
10
20
30
40
50
0 0.2 0.4 0.6 0.8 1Xgabapentin
Xg
ab
ap
en
tinx
Inte
nsi
ty
Job's plot analysis of fluorescence titrations of 6 (10-4 M in DMSO) with gabapentin in water (exc = 350
nm) (em = 515 nm).
Fluorimetric titrations of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)benzene 6
in DMSO with creatine and L-carnitine:
0
10
20
30
40
50
60
70
80
90
450 500 550 600 650 700
Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0005 0.0010 0.0015 0.0020 0.0025 0.0030
0
10
20
30
40
50
60
70
80
90Data: Data1_BModel: complejo 1:1 Chi^2 = 1.70395R^2 = 0.99641 Ao 78.93344 ±0.38829Af 5.03749 ±1.11732Co 0.0001 ±0K 2130.14674 ±90.24296
Inte
nsity
(a.
u.)
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4M in DMSO) with creatine in water (exc =
428 nm). Right: Titration profile fitted to a 1:1 model (λmax= 557 nm) for the titration of 6 with creatine.
0 equiv
30 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
0
10
20
30
40
50
60
70
80
450 500 550 600 650 700
Wavelength (nm)
Inte
nsi
ty (
a.u
.)
0.0000 0.0005 0.0010 0.0015 0.0020 0.0025 0.0030
0
10
20
30
40
50
60
70
80
complex 2:1 Chi^2 = 0.87142R^2 = 0.99845 Co 0.0001 ±0K1 12584.13228 ±4321.36026K2 6244.93566 ±881.17155Ao 77.78475 ±0.70153Af 2.05354 ±0.65023b 573609.90847 ±117217.05574
Inte
nsity
c(M)
Left: Emission spectra for the fluorimetric titration of 6 (10-4 M in DMSO) with L-carnitine in water
(exc= 426 nm). Right: Titration profile fitted to a 2:1 model (λmax = 555 nm) for the titration of 6 with L-
carnitine.
Job's plot analysis of fluorescence titrations of 6 with L-carnitine
Job's plot analysis of fluorescence titrations of 6 with L-carnitine revealed a maximum at a 66% mole
fraction, in accord with the proposed 2:1 binding stoichiometry
0
10
20
30
40
50
60
70
0 0.2 0.4 0.6 0.8 1Xcarnitine
Xca
rnit
inex
Inte
nsi
ty
Job's plot analysis of fluorescence titrations of 6 (10-4 M in DMSO) with L-carnitine in water (exc = 350
nm) (em = 515 nm).
0 equiv
28 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
1H NMR titration experiments of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)-
benzene 6 (10-1 M solution in DMSO) with β-alanine in DMSO.
By addition of increasing amounts of β-alanine, the two urea NH signals at δ 8.8 and 6.7 move to lower
field and progressively disappear as the H-bonds are formed. The aromatic signals become more
complicate but their chemical shifts remain unchanged.
8 6 4 2 0
1H NMR titration of 6 (10-1 M solution in DMSO-d6) with β-alanine in DMSO-d6.
2.0 equiv 1.5 equiv 1.0 equiv 0.5 equiv 0.3 equiv 0.1 equiv 0.0 equiv
δ (ppm)
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
The blue emission: deconvolution of the emission bands after titration:
Deconvolution of emission bands
0
10
20
30
40
50
60
70
80
90
100
400 450 500 550 600 650 700
Wavelength (nm)
Inte
nsi
ty (
a.u
.)
mixture 366nm (22,3eq)
mixture 390nm (22,3eq)
BLUE Emission
YELLOW emission
0eq 390nm
0 eq. 390 nm: the original fluorescence spectrum at λexc = 390 nm of 6 (10-4 M in DMSO).
Mixture 390 nm (22.3 equiv): Fluorescence spectrum at λexc = 390 nm after addition of 22.3 equivalents
of L-carnitine to the previous solution.
Mixture 366 nm (22.3 equiv): Fluorescence spectrum at λexc = 366 nm of the same mixture used in the
acquisition of the previous spectrum.
BLUE Emission: Fluorescence spectrum of the blue emission obtained by substraction of the yelow
emission spectrum at λexc = 390 nm from the bluish emission spectrum obtained at λexc = 366 nm from the
mixture of 6 and 22.3 equivalents of the analite (λmax = 475 nm).
YELLOW Emission: Fluorescence spectrum of the yellow emission obtained by substraction of the blue
emission spectrum at λexc = 366 nm from the emission spectrum obtained at λexc = 366 nm from the
mixture of 6 and 22.3 equivalents of the analite.
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Calculations:
All calculations were performed with the Gaussian 03 set of programs using the ONIOM method.1
Geometry optimizations were conducted using a two-level ONIOM procedure2, 3 in which the hybrid
functional B3LYP4, 5 along with the basis set 6-31G+(d,p) was used as the higher level of theory and the
semiempirical AM1 model6 was used as the lower level. After the optimization, analytical computations
of the Hessian matrix were performed to compute the nature of the located minima on the potential
energy surface. The real and model systems used in the two-layer ONIOM(MO:MO) calculations are
shown in the figure.
NH HN
NH HN
O O
N
N N
N
Computational ModelB3LYP/AM1
O O
R
References: 1 M. J. Frisch, G. W. Trucks, H. B. Schlegel, G. E. Scuseria, M. A. Robb, J. R. Cheeseman, J. J. A.
Montgomery, T. Vreven, K. N. Kudin, J. C. Burant, J. M. Millam, S. S. Iyengar, J. Tomasi, V. Barone, B.
Mennucci, M. Cossi, G. Scalmani, N. Rega, G. A. Petersson, H. Nakatsuji, M. Hada, M. Ehara, K. Toyota,
R. Fukuda, J. Hasegawa, M. Ishida, T. Nakajima, Y. Honda, O. Kitao, H. Nakai, M. Klene, X. Li, J. E.
Knox, H. P. Hratchian, J. B. Cross, C. Adamo, J. Jaramillo, R. Gomperts, R. E. Stratmann, O. Yazyev, A.
J. Austin, R. Cammi, C. Pomelli, J. W. Ochterski, P. Y. Ayala, K. Morokuma, G. A. Voth, P. Salvador, J.
J. Dannenberg, V. G. Zakrzewski, S. Dapprich, A. D. Daniels, M. C. Strain, O. Farkas, D. K. Malick, A.
D. Rabuck, K. Raghavachari, J. B. Foresman, J. V. Ortiz, Q. Cui, A. G. Baboul, S. Clifford, J. Cioslowski,
B. B. Stefanov, G. Liu, A. Liashenko, P. Piskorz, I. Komaromi, R. L. Martin, D. J. Fox, T. Keith, M. A.
Al-Laham, C. Y. Peng, A. Nanayakkara, M. Challacombe, P. M. W. Gill, B. Johnson, W. Chen, M. W.
Wong, C. Gonzalez, J. A. Pople, Gaussian 03, Revision C.02, Gaussian, Inc., Wallingford CT, 2004.
2 S. Humbel, S. Sieber, K. Morokuma, J. Chem. Phys. 1996, 105, 1959.
3 F. Maseras, K. Morokuma, J. Comput. Chem. 1995, 16, 1170.
4 A. D. Becke, J. Chem. Phys. 1993, 98, 5648.
5 C. T. Lee, W. T. Yang, R. G. Parr, Phys. Rev. B 1988, 37, 785.
6 M. J. S. Dewar, E. G. Zoebisch, E. F. Healy, J. J. P. Stewart, J. Am. Chem. Soc. 1985, 107, 3902.
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Optimized geometry of the 6:GABA·3DMSO complex at the ONIOM (B3LYP/6-31G*:AM1) in
GAUSSIAN 03.
Optimized geometry of the 6:PREGABALIN·3DMSO complex at the ONIOM (B3LYP/6-31G*:AM1) in
GAUSSIAN 03.
NH HNO OHN
CN
CN
NH
CN
NC
O O
NH3
3·[DMSO]
NH HNO OHN
CN
CN
NH
CN
NC
O O
NH3
3·[DMSO]
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
INTERFERENCE EXPERIMENTS:
We realized some interference experiments by adding 5 equivalents of interference compounds and then
succesive equivalents of the measured analyte (0.5 equiv, 1 equiv, 2 equiv, 3 equiv, 5 equiv, 7 equiv) and
represented in the plots the variation of the fluorescence intensity in each case.
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
6
crea
tine(
1 eq
uiv)
L-ca
rnitin
e(1
equiv
)
sarc
osin
e+cr
eatin
ine(5
equ
iv ea
ch)
(sar
cosin
e+cr
eatini
ne)+
crea
tine(
1 eq
uiv)
(S+C
t)+cr
eatin
e(1.5
equ
iv)
(S+C
t)+cr
eatin
e(2.0
equ
iv)
(S+C
t)+cr
eatin
e(3.0
equ
iv)
(S+C
t)+cr
eatin
e(5.0
equ
iv)
(sar
cosin
e+cr
eatini
ne)+
L-ca
rnitin
e(1 e
quiv)
(sar
c+cr
eatn
)+L-
carn
t(1.5
equ
iv)
(sar
c+cr
eatn
)+Lc
arnt
(2.0
equ
iv)
(sar
c+cr
eatn
)+Lc
arnt
(3.0
equ
iv)
(sar
c+cr
eatn
)+Lc
arnt
(5.0
equ
iv)
(sar
c+cr
eatn
)+Lc
arnt
(7.0
equ
iv)
1-I/I
o
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
6
Taurin
e+Glyc
ine+
Met
hionin
e(5
equiv
eac
h)
(T+G
+M)+
crea
tine(
1 eq
uiv)
(T+G
+M)+
crea
tine(
1.5
equiv
)
(T+G
+M)+
crea
tine(
2.0
equiv
)
(T+G
+M)+
crea
tine(
3.0
equiv
)
(T+G
+M)+
crea
tine(
5.0
equiv
)
(T+G
+M)+
Lcar
nitine
(1 e
quiv)
(T+G
+M)+
Lcar
nitine
(1.5
equ
iv)
(T+G
+M)+
Lcar
nitine
(2.0
equ
iv)
(T+G
+M)+
Lcar
nitine
(3.0
equ
iv)
(T+G
+M)+
Lcar
nitine
(5.0
equ
iv)
1-I
/Io
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Ultraviolet-visible titration of 1,3-bis[4-(1-dicyanomethyleneindan-5-yl)phenylureidomethyl)-
benzene 6 in DMSO with L-lysine in water:
lysine
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0.2
280 330 380 430 480
Wavelength (nm)
Ab
sorb
ance
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.2
1.4
1.6
2
2.5
3
3.5
4
5
400 nm
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0 1 2 3 4 5 6
equivalents of lysine
Ab
sorb
ance
Up: UV-visible absorption spectra for the colorimetric titration of 6 (10-5 M in DMSO) with L-lysine in
water. Down: Titration profile of the λmax = 400 nm for the UV-visible titration of 6 with L-lysine
0 equiv
5 equiv
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Titration Materials and Methods.
5×10-2 M, 5×10-3 M, 5×10-4 M solutions of every salt or aminoacid were prepared, then a 10-4 M solution of the compound
under study was prepared. For qualitative experiments, 2 mL solution of the compound under study were measured and the
corresponding amount of salt or aminoacid was added by micropipette.
Eppendorf Research micropipette characteristics:
MODEL Ep T.I.P.S. Volume Systematic error of measurement
Random error of measurement (CV)
2 - 20μL 2 - 200 2 μL ± 5.0 % ≤ 1.5 %
10 μL ± 1.2 % ≤ 0.6 %
20 μL ± 1.0 % ≤ 0.3 %
10 - 100μL 2 - 200 10 μL ± 3.0 % ≤ 1.0 %
50 μL ± 1.0 % ≤ 0.3 %
100 L ± 0.8 % ≤ 0.2 %
100 - 1000μL 50 - 1000 100 μL ± 3.0 % ≤ 0.6 %
5000 μL ± 1.0 % ≤ 0.2 %
1000 μL ± 0.6 % ≤ 0.2 %
500 - 5000μL 100 - 5000 500 μL ± 2.4 % ≤ 0.6 %
2500 μL ± 1.2 % ≤ 0.25 %
5000 μL ± 0.6 % ≤ 0.15 %
Quantitative studies:
As a general procedure, to a 5000 μl solution of complex, the corresponding amount (μL) of solution of the corresponding salt
(5×10-2 M, 5×10-3 M, 5×10-4 M solutions) was added, using the minor amount of solvent.
A table, corresponding to the number of equivalents and the corresponding added volumes is included:
Vtotal (µL) equiv C
5000 0 0
5005 0.05 4.995E-06
5010 0.1 9.98E-06
5015 0.15 1.4955E-05
5020 0.2 1.992E-05
5025 0.25 2.4876E-05
5030 0.3 2.9821E-05
5035 0.35 3.4757E-05
5040 0.4 3.9683E-05
5045 0.45 4.4599E-05
5050 0.5 4.9505E-05
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Vtotal (µL) equiv C
5055 0.55 5.4402E-05
5060 0.6 5.9289E-05
5065 0.65 6.4166E-05
5070 0.7 6.9034E-05
5075 0.75 7.3892E-05
5080 0.8 7.874E-05
5085 0.85 8.3579E-05
5090 0.9 8.8409E-05
5095 0.95 9.3229E-05
5100 1 9.8039E-05
5105 1.05 0.00010284
5110 1.1 0.00010763
5115 1.15 0.00011241
5120 1.2 0.00011719
5125 1.25 0.00012195
5130 1.3 0.00012671
5135 1.35 0.00013145
5140 1.4 0.00013619
5145 1.45 0.00014091
5150 1.5 0.00014563
5155 1.55 0.00015034
5160 1.6 0.00015504
5165 1.65 0.00015973
5170 1.7 0.00016441
5180 1.8 0.00017375
5190 1.9 0.00018304
5200 2 0.00019231
5210 2.1 0.00020154
5220 2.2 0.00021073
5230 2.3 0.00021989
5240 2.4 0.00022901
5250 2.5 0.0002381
5260 2.6 0.00024715
5270 2.7 0.00025617
5280 2.8 0.00026515
5300 3 0.00028302
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
Vtotal (µL) equiv C
5320 3.2 0.00030075
5340 3.4 0.00031835
5360 3.6 0.00033582
5380 3.8 0.00035316
5400 4 0.00037037
5405 4.5 0.00041628
5410 5 0.00046211
5415 5.5 0.00050785
5420 6 0.00055351
5430 7 0.00064457
5440 8 0.00073529
5450 9 0.00082569
5460 10 0.00091575
Supplementary Material (ESI) for Chemical CommunicationsThis journal is (c) The Royal Society of Chemistry 2011
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