Journal of Fluorescence

Design and Synthesis of a Pyridine Based Noncyclic Receptor as Pb2+ Ion Selective Probe
--Manuscript Draft-Manuscript Number: Full Title: Article Type: Keywords: Corresponding Author: JOFL-D-13-00211 Design and Synthesis of a Pyridine Based Noncyclic Receptor as Pb2+ Ion Selective Probe Short Communication Noncyclic receptor, Pb2+ Selective Probe, physicochemical changes, association constant, DFT. Anil S Kuwar, Ph.D., North Maharashtra University Jalgaon, Maharashtra INDIA

Corresponding Author Secondary Information: Corresponding Author's Institution: Corresponding Author's Secondary Institution: First Author: First Author Secondary Information: Order of Authors: Kundan C Tayade Anil S Kuwar, Ph.D., Umesh A Fegade Hemant Sharma Narinder Singh Umesh D Patil Sanjay B Attarde Order of Authors Secondary Information: Abstract: The Schiff's base 6,6'-((1E)-(pyridine-2,6diylbis(azanylylidene))bis(methanylylidene))bis(2-isopropyl-5-methylphenol) (receptor 3) has been designed and employed as fluorescent receptor for the ratiometric recognition of Pb2+ ion in organic-aqueous (0.68 mole fraction of DMF in water) media. The binding constant and physicochemical changes in receptor 3 upon addition of selected metal ions were investigated by fluorescence and absorption spectroscopic methods. Fluorescent receptor 3 shows a significant fluorescence enhancement in emission maxima (em) at 609 nm for an excitation wavelength (ex) of 304 nm with slit width 5 nm. The stoichiometry of the complex formed between receptor 3 and Pb2+ ion was found to be 1:1 (host:guest) which was established by Job's continuous variation method and confirmed by Density Functional Theory (DFT) & mass spectroscopy. The association constant (Ka) of receptor 3 with Pb2+ ion was computed with Benesi-Hildebrand plot using fluorescence and U. V. data and was found to be in concordance with each other. The calculated value of association constant was (5.2 0.1) x 103 M-1. Kundan C Tayade North Maharashtra University

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Manuscript Click here to download Manuscript: Main Document.doc Click here to view linked References 1 2 3 Design and Synthesis of a Pyridine Based Noncyclic Receptor as Pb2+ Ion Selective 4 Probe 5 a,b a Kundan C. Tayade , Anil S. Kuwar *, Umesh A. Fegadea,b, Hemant Sharmac, Narinder Singhc, Umesh D. 6 7 Patila, Sanjay B. Attardeb*. 8 9 a School of Chemical Sciences, North Maharashtra University, Jalgaon, 425001, (MS), India. 10 b 11 School of Environmental and Earth Sciences, North Maharashtra University, Jalgaon, 425001, (MS) , 12 13 India. 14 c 15 Department of Chemistry, Indian Institute of Technology, Ropar, Rupanagar, Punjab, India 16 * Corresponding authors. Tel.: +91-257-2257432; fax: +91-257-2257403; 17 18 E-mail addresses: kuwaras@gmail.com 19 20 -----------------------------------------------------------------------------------------------------------------------------------------21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65

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Design and Synthesis of a Pyridine Based Noncyclic Receptor as Pb2+ Ion Selective Probe
Abstract The Schiffs base 6,6'-((1E)-(pyridine-2,6-diylbis(azanylylidene))bis(methanylylidene))bis(2-

isopropyl-5-methylphenol) (receptor 3) has been designed and employed as fluorescent receptor for the ratiometric recognition of Pb2+ ion in organic-aqueous (0.68 mole fraction of DMF in water) media. The binding constant and physicochemical changes in receptor 3 upon addition of selected metal ions were investigated by fluorescence and absorption spectroscopic methods. Fluorescent receptor 3 shows a significant fluorescence enhancement in emission maxima (em) at 609 nm for an excitation wavelength (ex) of 304 nm with slit width 5 nm. The stoichiometry of the complex formed between receptor 3 and Pb2+ ion was found to be 1:1 (host:guest) which was established by Jobs continuous variation method and confirmed by Density Functional Theory (DFT) & mass spectroscopy. The association constant (Ka) of receptor 3 with Pb2+ ion was computed with Benesi-Hildebrand plot using fluorescence and U. V. data and was found to be in concordance with each other. The calculated value of association constant was (5.2 0.1) x 103 M-1. Keywords: Noncyclic receptor, Pb2+ Selective Probe, physicochemical changes, association constant, DFT. 1 Introduction In view of the original function of cations in a broad array of biological and chemical processes, the design and synthesis of receptors and chemosensors capable of selective binding and sensing of cations has attracted much attention in recent years [1]. Various receptors that have been reported for binding cations mostly utilize electrostatic interactions, noncovalent interaction, etc. to bind the guest species [2,3]. Noncyclic compounds containing plural bonding sites have gained considerable attention recently because of their ability of complexation towards ionic and /or neutral molecules [4,5]. We have so far synthesized non-cyclic compounds with plural hydroxyl groups and imine functional groups have been proved to serve not only as cation sensors but also as precursors for constructing supramolecular system, e.g. metallosupramolecular complexes. So the particular propose and efficient synthesis of noncyclic compound has continued to play a very important role in the area of supramolecular chemistry. Although many kinds of noncyclic compounds are prepared, development of a cost effective synthetic route under soft conditions to this type of compounds still remains a well-dressed and challenging subject for synthetic chemists [6,7]. Exposure to heavy metals, even at very trace level, is believed to be a serious health threat to flora and fauna. Among these metals lead has been considered most hazardous as it affects directly on nervous

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system, reproductive system and kidneys [8]. Up to now very few selective chemosensors have been developed for Pb2+ ions [9]. The probable reason for the rare synthesis was illustrated by the quenching of fluorescence by Pb2+ ions, since quenching was found not only detrimental for a high signal output during detection but also found unwanted for analytical purposes [10]. With lead the quenching may result from enhanced spinorbital coupling [11] or transfer of energy or electron [12,13]. Herein, the importance of our synthesised chemosensor which did not quench the fluorescence during recognition of Pb2+ ions was revealed. As a part of our work in the molecular recognition, we reported here design and synthesis of novel chemosensor 6,6'-((1E)-(pyridine-2,6-diylbis(azanylylidene))bis(methanylylidene))bis(2-isopropyl-5-

methylphenol) ( receptor 3) and studied its inclusion phenomenon with various cations. 2 Experimental All the required chemicals were purchased from Sigma Aldrich Ltd and used without further purification. The commercial grade solvents were used for analysis. For UVVis and fluorescence study the metal ions Cr3+, Mn2+, Fe3+, Co2+, Ni2+, Cu2+, Zn2+, Hg2+ were added as their chlorides, Bi2+, Pb2+, Ag+, Cd2+, Th4+ were added as their nitrates, Nd3+ was added as its oxide, Ce4+ was added as its sulphate while U6+ was added as its acetate. Fresh stock solutions of metal salts were prepared in DMF/H2O (90:10, v/v, c = 1x10-3 M) for analysis with receptor 3. Stock solutions of receptor 3 was freshly prepared in DMF/H2O (90:10, v/v, c = 1x10-4 M). The excitation wavelength (ex) was determined on UV-2450, UV-Vis Spectrophotometer, Shimadzu. Fluorescence studies were carried out on a HORIBA JOBIN YVON, Fluoromax-4 Spectrofluorometer. The solvent ration DMF/H2O (90:10, v/v) for receptor 1 was kept constant during the titrations. The structural determinations were carried out by using FT-IR, 1H-NMR,
13

C-NMR and LC MS spectroscopic techniques. FT-IR spectra of the synthesized receptor 3 was recorded

(Spectrophotometer Simadzu Asia Pacific Ptd. Ltd, Singapore) by uniformly mixing sample with KBr. 1H NMR spectrum was recorded in CDCl3 as a solvent (Varian Mercury YH-300 MHz Spectrometer) with tetramethylsilane (TMS) as an internal reference standard. Mass spectra were recorded under ESI mode, on Thermo Finnigan (model- LCQ Advantage MAX) mass spectrometer. 2.1 Synthesis of 6,6'-((1E)-(pyridine-2,6-diylbis(azanylylidene))bis(methanylylidene))bis(2-isopropyl5-methylphenol) (receptor 3) A solution of 2,6-diaminopyridine (0.11 g, 1 mmol) was added to a solution 2-hydroxy-3isopropyl-6-methyl-benzaldehyde (0.36 g, 2 mmol) in ethanol (50 mL) and mixture was refluxed for 12 hrs. There by deposited orange crystals of receptor 3 and was filtered, washed with petroleum ether (2 x 5 mL) and recrystallised from ethanol, 75 %, m.p. 142-144 0C.

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Analytical data for receptor 3: Yield: 75 %; FT-IR (KBr): = 3300-3050, 3039, 1597, 758, 653 cm-1; 1H NMR (300 MHz, CDCl3): = 1.26-1.29 (d, J=7.2 Hz, 12H, 4CH3), 2.57 (s, 6H, 2CH3), 3.37-3.46 (m, 2H, 2CH), 6.69-6.72 (d, J=7.5 Hz, 2H, 2Ar-H), 7.23-7.26 (d, 4H, 4Ar-H), 7.81-7.86 (t, J=7.5 Hz, 1H, Py-H), 9.97 (s, 2H, 2CH=N), 14.62 (bs, 2H, 2OH); 13C NMR (75 MHz, CDCl3): = 14.1, 25.3, 31.3, 100.5, 109.2, 114.5, 120.8, 122.6, 127.6, 130.3, 139.8, 147.6, 178.1; MS (ESI): Found m/z (%) = 430.22 (100), Calculated m/z = 429.24. 2.2 Determination of the binding constant by spectrofluorometric and UV titration The fluorescence titration experiment was conducted on HORIBA JOBIN YVON, Fluoromax-4 Spectrofluorometer in DMF/H2O (90:10) solvent system at room temperatures (298 K), for determination of association constant (Ka) of the receptor 3.Pb2+ ion complex formation. The titration experiment was endowed through a stepwise addition of changeable amount of Pb2+ ion solution (c = 1 x 10-3 M) to a solution of receptor 3 (2 mL, c = 1 x 10-4 M) in DMF/H2O in the cell. This was followed by a 5 min interval (in which the reaction cell was occasionally shaken to ensure mixing). Then, the fluorescence data were collected and processed using Benesi-Hildebrand methodology to calculate the association constant (Ka) of the metal ion for which selective fluorescence enhancement was obtained (here, Pb2+ ion). Similarly, association constant (Ka) was computed from the UV titration data by using the same methodology. In case of UV titration, the experiment was carried through a stepwise addition of changeable amount of Pb2+ ion solution (c = 1 x 10-6 M) to a solution of receptor 3 (2 mL, c = 1 x 10-8 M) in DMF/H2O in the cell. 3 Results and Discussion 2-hydroxy-3-isopropyl-6-methyl-benzaldehyde (1) was synthesized by the reported method of Kuwar et.al [14]. Schiffs base receptor 3 was prepared by condensation of two moles of 2-hydroxy-3isopropyl-6-methylbenzaldehyde (1) with one mole of 2,6-diaminopyridine (2) in ethanol (Scheme 1). Synthesized receptor 3 was characterized by FT-IR, 1H-NMR,
13

C NMR spectroscopy and LC-MS. The

spectral data obtained were found to be consistent with the structure of designed receptor 3.

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a
N NH2 OH HO

2
OH

H2N

Scheme-1 Synthesis of receptor 3 (a) C2H5OH, stirred, and refluxed 12 hrs.

The absorption spectrum of receptor 3 with Pb2+ ion was shown in Fig. 1. Receptor 3 exhibited absorption at 304, 328 and 356 nm in DMF/H2O (9:1, v/v) solution together with a swelling at 273 nm and a shoulder at 310 nm. The longer wavelength band at 365 nm and shoulder at 388 nm may be assigned to transitions associated with phenol substituted ring (Fig. 1). This observation clearly indicated towards a charge transfer from nitrogen (donor) of imine to Pb2+ (acceptor) metal ion. On the other hand, titration of receptor 3 with Pb2+ ion, resulted in saturation at 200 L with no further distinct fluorescence enhancement or quenching (Fig. 2). The plot of the absorbance verses [Pb2+] at 304 nm (Fig. 2, inset) shows a typical linear profile for formation of strong 1:1 complexation.

Fig. 1 Absorption spectrum of receptor compound 3 (c = 1 10-8 M, 2000 L) with Pb2+ ion (c= 1 10-6 M, 100 L).

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Fig. 2 UV titration spectrum of receptor compound 3 (c = 1 10-8 M) with Pb2+ ion (c = 1 10-6 M).

Fluorescence study was used as an important tool for studying the inclusion phenomenon of selected metal ions. The fluorescence response given by receptor 3 in DMF/water system (90:10 v/v, 0.68 mole fraction) towards the metal ions was studied. In the organic-aqueous solution of DMF and water, water was used for homogenization of receptor 3 and the guest metal ion. Fig. 3 illustrates changes in fluorescence of receptor 3 upon addition of aqueous solution of selected metal ions. With an excitation wavelength (ex) at 304 nm a distinct fluorescence enhancement in an emission peak with wavelength maxima (em) of receptor 3 at 609 nm was observed. Enhancements and decrease in height of an emission peak of receptor 3 upon addition of fixed amount (100 L) of various metal ions were recorded. It was revealed that receptor 3 (Fig. 3), was sensitive and selective towards Pb2+ ion showing distinct fluorescence enhancement.

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Fig. 3 Fluorescence spectra of receptor 3 (c = 1 10-4 M) prepared in DMF/H2O (9:1, v/v) upon addition of 100 L of respective metal ions (c = 1 10-3M) in aqueous phase for excitation wavelength at 304 nm.

The hydroxyl group works as an effective coordination sites for noncovalent interaction with the guest metal ions, especially transition series metal ions [15]. The importance of the phenolic -OH groups for inclusion of Pb2+ could be easily revealed from the binding pattern of receptor 3. Therefore, it appears that the coordination sites for adapting the Pb2+ in the cavity of receptor 3 were nitrogen of azomethine linkages and the phenolic OH groups. The competition experiments were conducted in presence of 1 equiv. of Pb2+ mixed with excess different metal ions (2 equiv.) and fluorescence profile of receptors 3 was almost not affected (Fig.4a and 4b). These results indicate that receptor 3 shows a good sensitivity and selectivity towards Pb2+ ion respectively over other competitive metal ions. The resulting data will be helpful for the researcher working in domains of biology and separation sciences.

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Fig. 4a Ratiomeric fluorescence response (F0-F/F0) of receptor 3 (c = 1 x 10-4 M) in DMF /H2O (9:1, v/v) upon addition of respective aqueous metal salt solutions (c = 1 x 10-3 M)

Fig. 4b Interference of respective metals in Pb2+ ion detection with receptor 3.

Progressive gradual fluorescence enhancement in the emission peak of receptor 3 centered at 609 nm on adding changeable amounts of Pb2+ ion solution was illustrated in Fig.5. On titrating aqueous solution of Pb2+ ions with receptor 3 prepared in DMF/H2O (9:1; v/v) a progressive fluorescence enhancement observed reaches the saturation point at 200 L. These observations can be attributed to complete complexation. Further increased concentration of Pb2+ ions or the presences of water do not result into any quenching of fluorescence. The detection limit (LOD) and limit of quantification (LOQ) is

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reasonably estimated to be 4.99 107 M and 1.51 10-6 M based on the ICH Q2B recommendations, below which the calibration graph is non linear[16]. LOD =3.3 /S LOQ =10 /S Where, S and represents the slope and the standard deviation of the intercept of regression line of the calibration curve.

Fig. 5 Fluorescence spectra of 3 (c = 1 10-4 M) in DMF/H2O (9:1, v/v) solution upon addition of increasing amount of Pb2+ (c = 4 10-3M) from 0-220 L with an excitation wavelength at 304 nm.

The stoichiometry of complexation of receptor 3 with Pb2+ was studied using Jobs continuous variation method [17,18]. Jobs plot and normalised plot obtained from fluorescence measurements showed the formation of receptors 3.Pb2+ complexes in 1:1 ligand to metal stoichiometric ratio (Fig. 6). This data was further confirmed by mass spectroscopy analysis. LCMS data showed the formation 1:1 complex between two deprotonated ligand (receptor 3) and a metal ion ([receptor 3.Pb2++2Na+] MW = 682.27; calcd for [C27H29N3O2Pb2++2Na], 681.20).

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Fig. 6 Fluorescence Job plot of receptor 3 with Pb2+ ion solution, [H] = [G] = 1 10-4 M

Using the Benesi-Hildebrand[19,20 ] (Eq.1) and (Eq.2) methodologies, we made the calculation of binding constant (Ka). 1 F-F0 1 A-A0 Where, F = F F0 = differential fluorescence intensities F = Fluorescence intensity during titration of metal ion with receptor. F0 = Fluorescence intensity of free receptor F = Fluorescence intensity after titration. A0 = Absorbance of free receptor A = Absorbance of receptor 3 during titration with metal ion From Benesi-Hildebrand equation, computed association constant (Ka) from fluorescence and UV data was Ka= 5.142 103 M-1 and 5.213 103 M-1 respectively. Fig. 7 illustrated plot of adjusted BenesiHildebrand plot of 1/F verses 1/[G] and 1/A verses 1/[G], where (A = A0 A). 1 (F-F0) K [G] 1 (A-A0) K [G] 1 (F-F0) 1 (A-A0) (Eq. 1) (Eq. 2)

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Fig. 7 Benesi-Hildebrand Plot from fluorescence of 1/F versus 1/[G] , Ka = Intercept/slope = 5.142 103 M-1 and from UV of 1/A versus 1/[G] , Ka = Intercept/slope = 5.213 103 M-1

The computational study was carried out by using Density Functional Theory (DFT) in an attempt to better understand the nature of receptor and its interaction with metal ion. The B3LYP function was employed for calculations with 6-31G/LanL2DZ basis set on Gaussian 09 programmer. The receptor 3 has three dimensional structure and B3LYP/6-31G basis set was employed for its optimization. In order to reduce the stress from steric crowding, two arms are in different direction as shown in Fig. 8a. From Fig. 8a, it was observed that nitrogen atoms constitute the pseudo cavity for metal ion. However, 3.Pb2+ was optimized by using B3LYP/ LanL2DZ basis set (Fig. 8b). On coordination of Pb2+, there is increase in the stability of the whole system which confirmed from value of energy optimization (Table 1). The gap between HOMO-LUMO is less for 3.Pb2+ as compare to receptor 3 alone as depicted in (Fig. 9). Further, the HOMO-LUMO of the 3.Pb2+ showed that LUMO is main contributor in electronic transition [21,22].

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Fig. 8 The DFT optimized structure of receptor 3 (A), and (B) 3.Pb2+ calculated at the B3LYP/6-31G and B3LYP/ LanL2DZ level respectively. The red, blue, gray, and dark gray spheres refer to O, N, C, and Pb 2+ atoms respectively.

Fig. 9 The HOMO-LUMO gap of: (A) receptor 3, and (B) 3.Pb2+ calculated at the B3LYP/6-31G and B3LYP/ LanL2DZ level respectively.

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Table 1: An optimized energy value calculated at B3LYP/6-31G/ LanL2DZ level.

Receptor 3 Energy (a.u.) 0.1493

3.Pb2+ 0.0343

4 Conclusions In summary, we designed and synthesised a noncyclic fluorescent receptor 3, which is selective and sensitive towards Pb2+ ion, in organic-aqueous medium. The presence of Pb2+ ion gave rise to distinct fluorescent enhancement and resulted in colour change which can be easily detected by the naked eye under UV irradiation. Furthermore, interference of other ions in Pb2+ ion detection with the receptor 3 was found negligible. The Stoichiometry for the host guest complex formation realized from the Jobs continuous variation method was 1:1 and the association constant (Ka) value obtained from fluorescence and UV titration data using Benesi-Hildebrand plot was Ka= 5.142 103 M-1 and 5.213 103 M-1 respectively which were found to be in harmony with each other. Acknowledgements ASK gratefully acknowledge financial support from the Department of Science and Technology (DST under Fast Track Scheme-SR/FT/CS-160/2011), Government of India, New Delhi.

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References 1. Bryce MR, Moore AJ, Hasan M, Ashwell GJ, Fraser AT, Clegg W, Hursthouse MB, Karaulov AI (1990) Angew Chem Int Ed Engl 29:1450-1452. 2. Sanz D, Perona A, Claramunt RM, Elguero J (2005) Tetrahedron 61:145-154. 3. Kilic A, Tas E, Deverec B, Yilmaz I (2007) Polyhedron 26:4009-4018. 4. Faridbod F, Ganjali MR, Dinarvand R, Norouzi P, Riahi S (2008) Sensors 8:1645-1703. 5. Taylor HE (2000) In Inductively Coupled Plasma Mass Spectrometry: Practices and Techniques. Academic Press, San Diego, CA. 6. Kuwar AS, Fegade UA, Tayade KC, Patil UD, Puschmann H, Gite VV, Dalal DS, Bendre RS (2013) J Fluoresc doi: 10.1007/s10895-013-1223-9. 7. Keiji H (2001) J Inclusion Phenom Macrocyclic Chem 39:193-209. 8. Chen CT, Huang WP (2002) J Am Chem Soc 124:6246-6247. 9. Lee KM, Chen XQ, Fang W, Kim JM, Yoon JY (2011) Macromol Rapid Commun 32:497-500. 10. Rurack K, Resch-Genger U, Rettig W (1998) J Photochem Photobiol A 118:143-149. 11. Mcclure DS (1952) J Chem Phys 20:682-686. 12. Varnes AW, Dodson RB, Whery EL (1972) J Am Chem Soc 94:946-950. 13. Lehn JM (1995) Supramolecular ChemistryConcept and Perspective. VCH, Weinheim. 14. Kuwar AS, Shimpi SR, Mahulikar PP, Bendre RS (2006) J Sci Ind Res 65:665-669. 15. Singh N, Kaur N, Mulrooney RC, Callan JF (2008) Tetrahedron Lett 49:6690-6692. 16. Liu XY, Bai DR, Wang S (2006) Angew Chem 118:5601-5604. 17. Job P (1928) Ann Chim Appl 9:113203. 18. Ghosh K, Kar D (2011) Beilstein J Org Chem 7:254-264. 19. Benesi HA, Hildebrand JH (1949) J Am Chem Soc 71:2703-2707. 20. Li Q, Guo Y, Xu J, Shao S (2011) J Photochem Photobiol B 103:140-144. 21. Becke AD (1993) J Chem Phys 98:5648-5652. 22. Lee C, Yang W, Parr RG (1988) Phys Rev B 37:785-789.

Figure Click here to download Figure: Scheme 1.docx

a
N NH2 OH HO

2
OH

H2N

Scheme-1 Synthesis of receptor 3 (a) C2H5OH, stirred, and refluxed 12 hrs.

Figure Click here to download Figure: Fig 1.docx

1.8 1.6 Host Host + Pb (II)

1.4
Absorbance 1.2 1 0.8 0.6 0.4 0.2 0 240 290 340 390 Wavelength (nm) 440 490

540

Fig. 1 Absorption spectrum of receptor compound 3 (c = 1 10-8 M, 2000 L) with Pb2+ ion (c= 1 10-6 M, 100 L).

Figure Click here to download Figure: Fig 2.docx

Fig. 2 UV titration spectrum of receptor compound 3 (c = 1 10-8 M) with Pb2+ ion (c = 1 10-6 M).

Figure Click here to download Figure: Fig 3.docx

240 220 200 180 Intensity ( 10000) ( a.u.) 160 140 Hg2+ 120 100 80 60 40 20 0 595 605 Wavelength (nm) Fig. 3 Fluorescence spectra of receptor 3 (c = 1 10-4 M) prepared in DMF/H2O (9:1, v/v) upon addition of 100 L of respective metal ions (c = 1 10-3M) in aqueous phase for excitation wavelength at 304 nm. 615 625 Bi3+ Cd2+ Ce4+ U6+ Ag+ Th4+ Nd3+ Pb2+ Host Cr3+

Mn2+
Co2+ Cu2+

Fe3+
Ni2+ Zn2+

Figure Click here to download Figure: Fig 4.docx

250 Fluorescence Intensity (a.u.) 200 150 100 50 0

Guest Metal Ions Fig. 4a Ratiomeric fluorescence response (F0-F/F0) of receptor 3 (c = 1 x 10-4 M) in DMF /H2O (9:1, v/v) upon addition of respective aqueous metal salt solutions (c = 1 x 10-3 M)

Competing ions Fluorescence Intensity (a.u.) Competing ions + Lead ion 300 200 100 0

Different Species

Fig. 4b Interference of respective metals in Pb2+ ion detection with receptor 3.

Figure Click here to download Figure: Fig 5.docx

Fig. 5 Fluorescence spectra of 3 (c = 1 10-4 M) in DMF/H2O (9:1, v/v) solution upon addition of increasing amount of Pb2+ (c = 4 10-3M) from 0-220 L with an excitation wavelength at 304 nm.

Figure Click here to download Figure: Fig 6.docx

5.00E-04 4.50E-04 4.00E-04 3.50E-04 [HG] 3.00E-04 2.50E-04 2.00E-04 1.50E-04 1.00E-04 5.00E-05 0.00E+00 0 0.1 0.2 0.3 0.4 0.5 Xi Fig. 6 Fluorescence Job plot of receptor 3 with Pb2+ ion solution, [H] = [G] = 1 10-4 M 0.6 0.7 0.8 0.9 1 1.1

Figure Click here to download Figure: Fig 7.docx

0.08 0.07 0.06 0.05 1/F

y = 7E-07x - 0.0036 R = 0.9969

16 14 12 10 1/A 8 6 4

y = 0.0001x + 0.5213 R = 0.9994

0.04
0.03 0.02 0.01 0 0 50000 1/[G] 100000 150000

2
0 0 20000 40000 60000 80000 100000 120000 1/[G]

Fig. 7 Benesi-Hildebrand Plot from fluorescence of 1/F versus 1/[G] , Ka = Intercept/slope = 5.142 103 M-1 and from UV of 1/A versus 1/[G] , Ka = Intercept/slope = 5.213 103 M-1

Figure Click here to download Figure: Fig 8.docx

Fig. 8 The DFT optimized structure of receptor 3 (A), and (B) 3.Pb2+ calculated at the B3LYP/6-31G and B3LYP/ LanL2DZ level respectively. The red, blue, gray, and dark gray spheres refer to O, N, C, and Pb 2+ atoms respectively.

Figure Click here to download Figure: Fig 9.docx

Fig. 9 The HOMO-LUMO gap of: (A) receptor 3, and (B) 3.Pb2+ calculated at the B3LYP/6-31G and B3LYP/ LanL2DZ level respectively.

Table Click here to download Table: Table 1.docx

Table 1: An optimized energy value calculated at B3LYP/6-31G/ LanL2DZ level.

Receptor 3 Energy (a.u.) 0.1493

3.Pb2+ 0.0343