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Supporting Information
A new pyrene based highly sensitive fluorescence probe for copper(II) and fluoride with living cell application
Shyamaprosad Goswamia Shampa Chakrabortya Sima Paula Sandipan Halderb Sukanya Panjac Subhro
Kanti Mukhopadhyayc
Department of Chemistry Bengal Engineering and Science University Shibpur Howrah 711103 West Bengal India E-mail spgoswamicalyahoocom Fax +91-3326682916
b Department of Chemistry Indian Institute of Technology Kanpur 208016 India
c Department of Microbiology The University of Burdwan Golapbag Burdwan-713104 India
TABLE OF CONTENTS
Sl No Contents Page No
1 General procedure for drawing Jobrsquos plot by fluorescence method 2
2 UVndashvis absorption spectra of receptor 2
3 Determination of Detection Limit 3
4 Determination of Association constant 3
5 Determination of fluorescence quantum yield 4
6 Fluorescence vs pH plot 5
7 1H NMR and ESI-MS spectra of compound (1) 6
8 1H NMR ESI-MS and FT-IR spectra of receptor and receptor + Cu 8
9 Fluorescence titration spectra of receptor with different metals 12
10 Fluorescence titration spectra of receptor with different anions 15
11 Fluorescence titration spectra of receptor with copper fluoride 18
12 Fluorescence titration spectra of receptor with Cu2+ and F-
respectively and vice versa 19
13 Biological study 19
14 References 20
Electronic Supplementary Material (ESI) for Organic This journal is copy The Royal Society of Chemistry 2014
2
1 General procedure for drawing Jobrsquos plot by fluorescence method
Stock solution of same concentration of sensor and Cu were prepared in the order of asymp 10 x 10-5 ML-1
CH3CN-H2O (73 vv at pH 75) media The fluorescence in each case with different hostndashguest ratio but
equal in volume was recorded Jobrsquos plots were drawn by plotting ∆IXhost vs Xhost (∆I = change of
fluorescence intensity of the spectrum during titration and Xhost is the mole fraction of the host in each case
respectively)
Figure (a) Figure (b)
Figure S1 Jobrsquos plot diagram of the (a) Cu2+-complex for BMPA (b) F--complex for BMPA determined by fluorescence method in CH3CN (where Xh is the mole fraction of the host and DI is the change of emission intensity)
2 UVndashvis absorption spectra of receptor
250 300 350 400 450 500000
005
010
015
020 Cu2+
Abso
rban
ce
Wavelength(nm)
Figure (a) Figure (b)
250 300 350 400 450000
002
004
006
008F
-
Abso
rban
ce
Wavelength (nm)
3
Figure S2 (a) UVndashvis absorption spectra of receptor (BMPA) (c = 1X 10-5 ML-1) upon gradual addition of Cu2+ (c = 2X 10-4 ML-1) in CH3CN (b) UVndashvis absorption spectra of receptor (BMPA) (c = 1X 10-5 ML-1) upon gradual addition of F- (c = 2X 10-4 ML-1) in CH3CN
3 Determination of Detection Limit
The detection limit DL of L for Cu+2 was determined from the following equation1
DL = K Sb1S Where K = 2 or 3 (we take 3 in this case) Sb1 is the standard deviation of the blank
solution S is the slope of the calibration curve From the graph we get slope = 3E+07 and Sb1 value is
1212 Thus using the formula we get the Detection Limit = 121 microM ie L can detect Cu2+ in this minimum
concentration
From the graph we get slope = 5E+07 and Sb1 value is 48558 Thus using the formula we get the Detection
Limit = 291 microM ie L can detect F- in this minimum concentration
4 Determination of Association constant
The spectra of these solutions were recorded by means of fluorescence methods Binding constant was
calculated according to the Benesi-Hildebrand equation Ka was calculated following the equation stated
below
1(F-Fo) = 1K(FmaxndashFo) [Cu2+]n + 1[Fmax-Fo]
1(F-Fo) = 1K(FmaxndashFo) [F-]n + 1[Fmax-Fo]
Here Fo is the fluorescence of receptor in the absence of guest F is the fluorescence recorded in the presence
of added guest Fmax is fluorescence in presence of added [Cu2+]max and [F-]max K is the association
constant (M-1) The association constant (Ka) could be determined from the slope of the straight line of the
plot of 1(F-Fo) against 1[Cu2+]n and 1[F-]n respectively The association constant (Ka) as determined by
4
fluorescence titration method for sensor with Cu2+ and F- is found to be 10times105 M-1 and 107x105 M-1
respectively
Figure (a) Figure (b)
Figure S3 BenesindashHildebrand plot from Fluorescence titration data of receptor BMPA (1x10-5M) with (a)
Cu2+ and (b) F- respectively
5 Determination of fluorescence quantum yield
Here the quantum yield φ was measured by using the following equation φx = φs (Fx Fs)(As Ax)(nx2
ns2) where X amp S indicate the unknown and standard solution respectively φ = quantum yield F = area
under the emission curve A = absorbance at the excitation wave length n = index of refraction of the
solvent Here φ measurements were performed using anthracene in ethanol as standard [φ = 027] (error ~
10)
The quantum yield of BMPA itself is 088 which remarkably changed to 152 on the formation of a complex
with Cu2+ metal ion and 402 due to binding with F- respectively
5
6 Fluorescence vs pH plot
2 3 4 5 6 7 8 9 10
200
210
220
230
240
250
Fluo
resc
ence
(nm
)
PH
3 4 5 6 7 8 9
370
380
390
400
410
420
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9 10
185
190
195
200
205
210
215
220
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9400
410
420
430
440
450Fl
uore
scen
ce
PH
Figure S4 (a) Plots of fluorescence at 414 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 597 (b) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN-H2O (73 vv) medium in presence of Cu2+ (20 equiv 20 x10-4 M) at 414 nm (c) Plots of fluorescence at 448 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 624 (d) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN medium in presence of F- (10 equiv 20 x10-4 M) at 448nm
(a) (b)
(c) (d)
6
7 1H NMR and HRMS spectra of compound (1)
1H NMR of compound (1)
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
2
1 General procedure for drawing Jobrsquos plot by fluorescence method
Stock solution of same concentration of sensor and Cu were prepared in the order of asymp 10 x 10-5 ML-1
CH3CN-H2O (73 vv at pH 75) media The fluorescence in each case with different hostndashguest ratio but
equal in volume was recorded Jobrsquos plots were drawn by plotting ∆IXhost vs Xhost (∆I = change of
fluorescence intensity of the spectrum during titration and Xhost is the mole fraction of the host in each case
respectively)
Figure (a) Figure (b)
Figure S1 Jobrsquos plot diagram of the (a) Cu2+-complex for BMPA (b) F--complex for BMPA determined by fluorescence method in CH3CN (where Xh is the mole fraction of the host and DI is the change of emission intensity)
2 UVndashvis absorption spectra of receptor
250 300 350 400 450 500000
005
010
015
020 Cu2+
Abso
rban
ce
Wavelength(nm)
Figure (a) Figure (b)
250 300 350 400 450000
002
004
006
008F
-
Abso
rban
ce
Wavelength (nm)
3
Figure S2 (a) UVndashvis absorption spectra of receptor (BMPA) (c = 1X 10-5 ML-1) upon gradual addition of Cu2+ (c = 2X 10-4 ML-1) in CH3CN (b) UVndashvis absorption spectra of receptor (BMPA) (c = 1X 10-5 ML-1) upon gradual addition of F- (c = 2X 10-4 ML-1) in CH3CN
3 Determination of Detection Limit
The detection limit DL of L for Cu+2 was determined from the following equation1
DL = K Sb1S Where K = 2 or 3 (we take 3 in this case) Sb1 is the standard deviation of the blank
solution S is the slope of the calibration curve From the graph we get slope = 3E+07 and Sb1 value is
1212 Thus using the formula we get the Detection Limit = 121 microM ie L can detect Cu2+ in this minimum
concentration
From the graph we get slope = 5E+07 and Sb1 value is 48558 Thus using the formula we get the Detection
Limit = 291 microM ie L can detect F- in this minimum concentration
4 Determination of Association constant
The spectra of these solutions were recorded by means of fluorescence methods Binding constant was
calculated according to the Benesi-Hildebrand equation Ka was calculated following the equation stated
below
1(F-Fo) = 1K(FmaxndashFo) [Cu2+]n + 1[Fmax-Fo]
1(F-Fo) = 1K(FmaxndashFo) [F-]n + 1[Fmax-Fo]
Here Fo is the fluorescence of receptor in the absence of guest F is the fluorescence recorded in the presence
of added guest Fmax is fluorescence in presence of added [Cu2+]max and [F-]max K is the association
constant (M-1) The association constant (Ka) could be determined from the slope of the straight line of the
plot of 1(F-Fo) against 1[Cu2+]n and 1[F-]n respectively The association constant (Ka) as determined by
4
fluorescence titration method for sensor with Cu2+ and F- is found to be 10times105 M-1 and 107x105 M-1
respectively
Figure (a) Figure (b)
Figure S3 BenesindashHildebrand plot from Fluorescence titration data of receptor BMPA (1x10-5M) with (a)
Cu2+ and (b) F- respectively
5 Determination of fluorescence quantum yield
Here the quantum yield φ was measured by using the following equation φx = φs (Fx Fs)(As Ax)(nx2
ns2) where X amp S indicate the unknown and standard solution respectively φ = quantum yield F = area
under the emission curve A = absorbance at the excitation wave length n = index of refraction of the
solvent Here φ measurements were performed using anthracene in ethanol as standard [φ = 027] (error ~
10)
The quantum yield of BMPA itself is 088 which remarkably changed to 152 on the formation of a complex
with Cu2+ metal ion and 402 due to binding with F- respectively
5
6 Fluorescence vs pH plot
2 3 4 5 6 7 8 9 10
200
210
220
230
240
250
Fluo
resc
ence
(nm
)
PH
3 4 5 6 7 8 9
370
380
390
400
410
420
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9 10
185
190
195
200
205
210
215
220
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9400
410
420
430
440
450Fl
uore
scen
ce
PH
Figure S4 (a) Plots of fluorescence at 414 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 597 (b) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN-H2O (73 vv) medium in presence of Cu2+ (20 equiv 20 x10-4 M) at 414 nm (c) Plots of fluorescence at 448 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 624 (d) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN medium in presence of F- (10 equiv 20 x10-4 M) at 448nm
(a) (b)
(c) (d)
6
7 1H NMR and HRMS spectra of compound (1)
1H NMR of compound (1)
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
3
Figure S2 (a) UVndashvis absorption spectra of receptor (BMPA) (c = 1X 10-5 ML-1) upon gradual addition of Cu2+ (c = 2X 10-4 ML-1) in CH3CN (b) UVndashvis absorption spectra of receptor (BMPA) (c = 1X 10-5 ML-1) upon gradual addition of F- (c = 2X 10-4 ML-1) in CH3CN
3 Determination of Detection Limit
The detection limit DL of L for Cu+2 was determined from the following equation1
DL = K Sb1S Where K = 2 or 3 (we take 3 in this case) Sb1 is the standard deviation of the blank
solution S is the slope of the calibration curve From the graph we get slope = 3E+07 and Sb1 value is
1212 Thus using the formula we get the Detection Limit = 121 microM ie L can detect Cu2+ in this minimum
concentration
From the graph we get slope = 5E+07 and Sb1 value is 48558 Thus using the formula we get the Detection
Limit = 291 microM ie L can detect F- in this minimum concentration
4 Determination of Association constant
The spectra of these solutions were recorded by means of fluorescence methods Binding constant was
calculated according to the Benesi-Hildebrand equation Ka was calculated following the equation stated
below
1(F-Fo) = 1K(FmaxndashFo) [Cu2+]n + 1[Fmax-Fo]
1(F-Fo) = 1K(FmaxndashFo) [F-]n + 1[Fmax-Fo]
Here Fo is the fluorescence of receptor in the absence of guest F is the fluorescence recorded in the presence
of added guest Fmax is fluorescence in presence of added [Cu2+]max and [F-]max K is the association
constant (M-1) The association constant (Ka) could be determined from the slope of the straight line of the
plot of 1(F-Fo) against 1[Cu2+]n and 1[F-]n respectively The association constant (Ka) as determined by
4
fluorescence titration method for sensor with Cu2+ and F- is found to be 10times105 M-1 and 107x105 M-1
respectively
Figure (a) Figure (b)
Figure S3 BenesindashHildebrand plot from Fluorescence titration data of receptor BMPA (1x10-5M) with (a)
Cu2+ and (b) F- respectively
5 Determination of fluorescence quantum yield
Here the quantum yield φ was measured by using the following equation φx = φs (Fx Fs)(As Ax)(nx2
ns2) where X amp S indicate the unknown and standard solution respectively φ = quantum yield F = area
under the emission curve A = absorbance at the excitation wave length n = index of refraction of the
solvent Here φ measurements were performed using anthracene in ethanol as standard [φ = 027] (error ~
10)
The quantum yield of BMPA itself is 088 which remarkably changed to 152 on the formation of a complex
with Cu2+ metal ion and 402 due to binding with F- respectively
5
6 Fluorescence vs pH plot
2 3 4 5 6 7 8 9 10
200
210
220
230
240
250
Fluo
resc
ence
(nm
)
PH
3 4 5 6 7 8 9
370
380
390
400
410
420
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9 10
185
190
195
200
205
210
215
220
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9400
410
420
430
440
450Fl
uore
scen
ce
PH
Figure S4 (a) Plots of fluorescence at 414 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 597 (b) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN-H2O (73 vv) medium in presence of Cu2+ (20 equiv 20 x10-4 M) at 414 nm (c) Plots of fluorescence at 448 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 624 (d) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN medium in presence of F- (10 equiv 20 x10-4 M) at 448nm
(a) (b)
(c) (d)
6
7 1H NMR and HRMS spectra of compound (1)
1H NMR of compound (1)
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
4
fluorescence titration method for sensor with Cu2+ and F- is found to be 10times105 M-1 and 107x105 M-1
respectively
Figure (a) Figure (b)
Figure S3 BenesindashHildebrand plot from Fluorescence titration data of receptor BMPA (1x10-5M) with (a)
Cu2+ and (b) F- respectively
5 Determination of fluorescence quantum yield
Here the quantum yield φ was measured by using the following equation φx = φs (Fx Fs)(As Ax)(nx2
ns2) where X amp S indicate the unknown and standard solution respectively φ = quantum yield F = area
under the emission curve A = absorbance at the excitation wave length n = index of refraction of the
solvent Here φ measurements were performed using anthracene in ethanol as standard [φ = 027] (error ~
10)
The quantum yield of BMPA itself is 088 which remarkably changed to 152 on the formation of a complex
with Cu2+ metal ion and 402 due to binding with F- respectively
5
6 Fluorescence vs pH plot
2 3 4 5 6 7 8 9 10
200
210
220
230
240
250
Fluo
resc
ence
(nm
)
PH
3 4 5 6 7 8 9
370
380
390
400
410
420
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9 10
185
190
195
200
205
210
215
220
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9400
410
420
430
440
450Fl
uore
scen
ce
PH
Figure S4 (a) Plots of fluorescence at 414 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 597 (b) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN-H2O (73 vv) medium in presence of Cu2+ (20 equiv 20 x10-4 M) at 414 nm (c) Plots of fluorescence at 448 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 624 (d) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN medium in presence of F- (10 equiv 20 x10-4 M) at 448nm
(a) (b)
(c) (d)
6
7 1H NMR and HRMS spectra of compound (1)
1H NMR of compound (1)
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
5
6 Fluorescence vs pH plot
2 3 4 5 6 7 8 9 10
200
210
220
230
240
250
Fluo
resc
ence
(nm
)
PH
3 4 5 6 7 8 9
370
380
390
400
410
420
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9 10
185
190
195
200
205
210
215
220
Fluo
resc
ence
(nm
)
pH
3 4 5 6 7 8 9400
410
420
430
440
450Fl
uore
scen
ce
PH
Figure S4 (a) Plots of fluorescence at 414 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 597 (b) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN-H2O (73 vv) medium in presence of Cu2+ (20 equiv 20 x10-4 M) at 414 nm (c) Plots of fluorescence at 448 nm of BMPA as a function of pH The solid line is the best fit pH titration curve with pKa 624 (d) Absorbance intensity of BMPA (10 microM) at various pH values in CH3CN medium in presence of F- (10 equiv 20 x10-4 M) at 448nm
(a) (b)
(c) (d)
6
7 1H NMR and HRMS spectra of compound (1)
1H NMR of compound (1)
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
6
7 1H NMR and HRMS spectra of compound (1)
1H NMR of compound (1)
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
7
HRMS of compound (1)
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
8
8 1H NMR HRMS and FT-IR spectra of receptor and receptor + Cu(II)
1H NMR of receptor
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
9
13C NMR of receptor
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
10
HRMS spectra of receptor
(BMPA+Na++1)
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
11
HRMS spectra of receptor + Cu
FT-IR data of (a) receptor and (b) receptor + Cu
(a) (b)
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
12
9 Fluorescence emission spectra of receptor ( C = 1x 10-5 M in CH3CN H2O(73 vv at pH 75) with
different cations as Na+ K+ Ba2+ Ga3+ Mg2+ Ca2+ Mn2+ Fe2+ Ni2+ Co2+ Zn2+ Ag+ Cd2+ Hg2+ Pb2+
In3+ Cr3+ and Fe3+ in CH3CN H2O (C = 2x 10-4 M 73 vv at pH 75) The solutions of metal ions were
prepared from NaClO4 KClO4 Ba(ClO4)24H2O GaCl3 Mg(ClO4)26H2O Ca(ClO4)24H2O
Mn(ClO4)26H2O Fe(SO4)2 Ni(ClO4)26H2O Co(ClO4)26H2O Zn(ClO4)26H2O AgNO3 Cd(ClO4)2H2O
HgCl2 Pb(ClO4)2 InCl3 CrCl36H2O and FeCl3 respectively
330 360 390 420 450 480 5100
50
100
150
200
250
300 Na+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 K+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
0
50
100
150
200
250
300 Ga3+
Fluo
resc
ence
Wavelength(nm)
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
13
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mg2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ca2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Mn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ni2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Co2+
Fluo
resc
ence
Wavelength(nm)
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
14
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Ag+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Ba2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Zn2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Cd2+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Hg2+
Fluo
resc
ence
Wavelength(nm)
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
15
330 360 390 420 450 480 510
50
100
150
200
250
300
350 Pb2+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 In3+
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 5100
50
100
150
200
250
300 Cr3+
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
50
100
150
200
250
300
350 Fe3+
Fluo
resc
ence
Wavelength(nm)
10 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with different anions
as AcO- Cl- Br- I- BzO- SH- H2PO4- PO4
3- S2- N3- P2O7
4- SCN- ADP and ATP in MeOH H2O (C
= 2x 10-4 M) The AcO- Cl- Br- I- and BzO- as their tetra butyl salts SH- H2PO4- PO4
3- S2- N3-
P2O74- and SCN- as their sodium salts ADP and ATP as their potassium salts in CH3CN
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
16
300 350 400 450 500 5500
50
100
150
200
250 OAc-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Cl-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 Br-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 I-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
100
200
300
400 BzO-
Fluo
resc
ence
Wavelength(nm) 330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SH-
Fluo
resc
ence
Wavelength(nm)
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
17
330 360 390 420 450 480 510
100
200
300
400 H2PO4
-Fl
uore
scen
ce
Wavelength(nm) 330 360 390 420 450 480 510
100
200
300
400 PO43-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 S2-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 N-3
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 P2O74-
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 SCN-
Fluo
resc
ence
Wavelength(nm)
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
18
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ADP
Fluo
resc
ence
Wavelength(nm)
330 360 390 420 450 480 510
30
60
90
120
150
180
210
240 ATP
Fluo
resc
ence
Wavelength(nm) 11 Fluorescence emission spectra of BMPA (C = 1x 10-5 M in CH3CN) at pH 75 with Copper fluoride
300 350 400 450 500 5500
150
300
450
600
750
900
1050 CuF2
Abso
rban
ce(n
m)
Wavelength
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
19
12 Fluorescence titration spectra of receptor with Cu2+ and F- respectively and vice versa
350 400 450 500 5500
100
200
300
400
500
600
BMPA
Abso
rban
ce(n
m)
Wavelength
300 350 400 450 500 550
0100200300400500600700800900
1000
BMPAAbso
rban
ce(n
m)
Wavelength
Figure (a) Figure (b) Fig 7 (a) Fluorescence spectra of (a) [BMPA + Cu2+] upon addition of F- and (b) [BMPA + F-] upon addition of Cu2+ in CH3CN 13 Biological study Preparation of Tecoma stans pollen cells
Pollen grains were obtained from freshly collected mature buds of Tecoma stans a common ornamental
plant with bell shaped bright yellow flower by crashing stamens on a sterile Petri plate and suspending them
in normal saline After crushing the stamina debrishes are removed by filtering through a thin layer of non
absorbant cotton and the suspended pollens are collected by centrifugation at 3000 rpm for five minutes The
pollen pellet was then washed twice in normal saline and then incubated in a solution of copper perchlorate
salt (1 mgml) for 45 minutes at ambient temperature After incubation they are again washed in normal
saline as mentioned above and then treated with the receptor BMPA(1 mgml) solution for 45 minutes Then
the cells were washed with normal saline and photographed under various objectives using UV filter in a
Leica DM1000 Fluorescence microscope in presence and absence of the ligand Cells without treatment were
used as control
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
20
Preparation of Candida albicans cells
Candida albicans cells (IMTECH No 3018) from exponentially growing culture in Potato Dextrose Broth
medium (pH 60 incubation temperature 37 0C) were centrifuged at 3000 rpm for 8 minutes washed twice
with 01 M HEPES buffer at pH 74 Then it was treated with copper perchlorate salt (1 mgml) for 45
minutes After incubation the cells were washed again with HEPES buffer at pH 74 These salt treated cells
were treated with the receptor (1mgml) for 45 minutes at ambient temperature After incubation the cells
were washed again with HEPES buffer at pH 74 The cells were then mounted on grease free glass slide and
observed under a Leica DM1000 fluorescence microscope equipped with UV filter Cells without treatment
were used as control
14 References
1 M Zhu M Yuan X Liu J Xu J Lv C Huang H Liu Y Li S Wang D Zhu Org Lett 2008 10
1481
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