Exploring the Scope of Photo-Induced Electron TransferâChelation

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Article Cite This: ACS Omega 2018, 3, 4262−4275

Exploring the Scope of Photo-Induced Electron Transfer−ChelationEnhanced Fluorescence−Fluorescence Resonance Energy Transfer Processes for Recognition and Discrimination of Zn2+, Cd2+, Hg2+, and Al3+ in a Ratiometric Manner: Application to Sea Fish Analysis Milan Ghosh, Sabyasachi Ta, Mahuya Banerjee, Md Mahiuddin, and Debasis Das* Department of Chemistry, The University of Burdwan, Burdwan, 713104 West Bengal, India S Supporting Information *

ABSTRACT: A rhodamine-based smart probe (RHES) has been developed for tracelevel detection and discrimination of multiple cations, viz. Al3+, Zn2+, Cd2+, and Hg2+ in a ratiometric manner involving photo-induced electron transfer−chelation-enhanced fluorescence−fluorescence resonance energy transfer processes. The method being very fast and highly selective allows their bare eye visualization at a physiological pH. The optimized geometry and spectral properties of RHES and its cation adducts have been analyzed by time-dependent density functional theory calculations. RHES detects as low as 1.5 × 10−9 M Al3+, 1.2 × 10−9 M Zn2+, 6.7 × 10−9 M Cd2+, and 1.7 × 10−10 M Hg2+, whereas the respective association constants are 1.33 × 105 M−1, 2.11 × 104 M−1, 1.35 × 105 M−1, and 4.09 × 105 M−1. The other common ions do not interfere. The probe is useful for intracellular imaging of Zn2+, Cd2+, and Hg2+ in squamous epithelial cells. RHES is useful for the determination of the ions in sea fish and real samples.

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INTRODUCTION A smart probe that converts a highly selective molecular recognition of multiple ions into easily detectable signals is very attractive in the present scenario. The techniques to visualize bioactive and environment-relevant cations have immense importance in biomedical analysis and environmental monitoring.1,2 The techniques like inductively coupled plasma mass/atomic emission spectroscopy, atomic absorption spectroscopy,3 colorimetry,4 spectrophotometry,5−7 and voltammetry8 generally require sophisticated expensive equipment, tedious time-consuming sample preparation procedure, and trained skilled operator. In contrast, fluorescence spectroscopy is very useful to provide instantaneous detection, visual perception, and inexpensive methodology that excludes sample pretreatment. A single probe capable to sense multiple ions is cost-effective and highly desirable for practical applications.9 Particularly, a probe that selectively and specifically detects and discriminates elements belonging to the same group in the periodic table through the photo-induced electron transfer−chelation-enhanced fluorescence−fluorescence resonance energy transfer (PET−CHEF−FRET) processes in a ratiometric manner is highly demanding as well as difficult to achieve10 because it rules out the adverse environmental effects like pH, polarity, probe concentration, and excitation power on the emission intensity via built-in correction to the signal ratio of emission intensities at two different wavelengths.11−13 Although Al is extensively used in food packaging, cookware, antiperspirants, drinking water supplies, bleached flour, deodorants, antacids, manufacturing of cars, and computers,14−16 it may cause neurotoxicity, disorders of homeostasis, Parkinson’s © 2018 American Chemical Society

disease, Alzheimer’s disease (AD), amyotrophic lateral sclerosis (ALS), microcytic hypochromic anemia, myopathy, and anemia.17−22 It also inhibits plant growth by increasing the acidity of the soil.23 Zinc, the most biocompatible metal,24−27 is present in all forms of life28 and plays a vital role in numerous biological processes including brain activity, gene transcription, immune function, etc.29 Zn2+-based compounds are used as radioprotective agents,30 tumor photosensitizers,31 antidiabetic insulin-mimetic,32 antibacterial, antimicrobial, and anticancer agents.33 It also reduces cardio- and hepatotoxicity induced by some anticancer drugs.34 Its abnormal metabolism causes health problems like prostate cancer, delayed sexual maturation and impotence, type 2 diabetes mellitus, ALS, Wilson’s diseases, agerelated macular degeneration,35−37 AD,38 cerebral ischemia,39 epilepsy,40 and acrodermatitis enteropathica.41 On the other hand, Cd occupies the seventh position among the top 20 hazardous substances in the priority list prepared by the Agency for Toxic Substances and Disease Registry and the US Environmental Protection Agency.42 The World Health Organization allows a maximum of 3 ppb Cd2+ in drinking water.43 Cd released from Ni−Cd batteries, phosphate fertilizers, pigments, and semiconducting quantum dots/rods44 causes renal dysfunction, calcium metabolism disorders, reduced lung capacity, risk of cancer, and health disorders.45 Thus, the Received: February 13, 2018 Accepted: April 4, 2018 Published: April 16, 2018 4262

DOI: 10.1021/acsomega.8b00266 ACS Omega 2018, 3, 4262−4275

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ACS Omega determination of trace-level Cd2+ in environmental and biological samples has great significance. Finally, it is well-known that Hg damages kidney, skin, respiratory system, central nervous system, and other organs.46,47 It is observed in the literature that most of the reported probes relevant to these cations either detect one or dual cations.48−50 Moreover, very few fluorescence probes detect and discriminate Zn2+ and Cd2+. In most cases, Cd2+ interferes with Zn2+ sensing.51a A simple turn-on probe51b for the detection of both Zn2+ and Cd2+ suffers limitations such as narrow difference in emission wavelengths (λEm for Zn2+, 572 nm and λEm for Cd2+, 565 nm) and low detection limit. An amino-terpyridine-based probe51c detects both Zn2+ and Cd2+ with an incremental enhancement of emission intensity at 535 nm but fails to discriminate Zn2+ and Cd2+. Chen et al.51d have reported a turnon fluorescence probe for Zn2+, Cd2+, and Hg2+ that also fails to discriminate them. It is to be noted that the molecular arithmetic that converts a chemically encoded analysis (input) into an optical signal52,53 (output) is potentially an interesting research area in modern unconventional computing system54 that insisted us to construct a scientific logic gate using our present probe. Rhodamine derivatives possess excellent photophysical properties, viz. high quantum yield,55 photostability, bioavailability, excitation, and emission in the visible region,55 and undergo an equilibrium between the nonfluorescent “spirolactam ring” and fluorescent “ring-open” forms to allow analyte sensing through “off−on” switching.56 Herein, a smart probe is being reported that selectively detects and discriminates Al3+, Zn2+, Cd2+, and Hg2+, utilizing their significantly different emission wavelengths and eye-catching colors under ultraviolet (UV) and visible light. The ratiometric sensing mechanism involves PET−CHEF−FRET processes. The ability of RHES for imaging intracellular Zn 2+, Cd 2+ , and Hg2+ have been demonstrated in live cells.

green (Zn2+), sky blue (Cd2+), and intense bloodred (Hg2+) (λEx, 306 nm) (Figure 1). Additionally, for Al3+ and Hg2+, the solution

Figure 1. Colors of RHES (20 μM) in visible light (left) and under UV light (right) in the presence of Al3+, Zn2+, Cd2+, and Hg2+ (20 mM N-(2hydroxyethyl)piperazine-N′-ethanesulfonic acid (HEPES)-buffered MeOH/H2O (4/1, v/v, pH 7.4)).

looks pink and red in bare eye (Figure 1). Interestingly, out of all the common ions tested, the emission intensity enhances for Al3+ (51-fold along with 176 nm red shift from 397 to 573 nm), Zn2+ (29-fold along with 85 nm red shift from 397 to 482 nm), Cd2+ (36-fold along with 2 nm blue shift from 397 to 395 nm), and Hg2+ (89-fold along with 180 nm red shift from 397 to 577 nm) (Figure 2). The spectroscopic properties of RHES depend on the pH of the media as it contains pH-susceptible donor sites. Moreover, the biological application of RHES demands its efficiency at a physiological pH. Hence, RHES is mixed with Al3+, Zn2+, Cd2+, and Hg2+ in different sets at different pH values (pH 3.0−12.0). The difference in emission intensities between the free RHES and its metal ion adducts is highest near the physiological pH 7.4, and hence chosen for the entire study (Figure S2, Supporting Information). Figures 2 and 3a demonstrate the selectivity of RHES for Al3+ (λEm = 573 nm). Figure S3 (Supporting Information) indicates no interference from the common cations. Upon a gradual addition of Al3+ (from 0.005 to 1600 μM), the colorless RHES slowly turns sky blue, whereby the emission intensity increases at 362 nm. After 25 min, the emission intensity gradually increases at 573 nm. Further, with increasing time, the emission intensity at 362 nm remains unaltered, whereas it increases at 573 nm. It is proposed that the initial coordination of phenol −O and imine N to Al3+ is responsible for the weak emission at 362 nm. Subsequent coordination by the −O donors from the carbonyl functionality of the spirolactam moiety to Al3+ leads to fluorescence enhancement at 573 nm along with the opening of the spirolactam ring resulting in pink coloration57 (Figure S4, Supporting Information). The time-dependent color changes of the RHES−Al3+ system under a handheld UV lamp are presented in Figure S4 (Supporting Information). Figure 3b shows the fluorescence titration of RHES by Al3+. RHES detects as low as 1.5 × 10−9 M Al3+ (Figure S5, Supporting Information). Figure 3c shows the [Al3+] versus emission intensity plot at 573 nm (red balls) and 362 nm (inset, sky blue balls). The red shift of the emission band is accompanied by a 51-fold fluorescence enhancement (λEm, 573 nm; Φ, 6.6-fold enhancement). Moreover, in the presence of Al3+, 1.5-fold increase of F573/ F362 is observed (Figure S6, Supporting Information). The plot of fluorescence enhancement versus [Al3+] at different wavelengths is shown in Figure S6 (Supporting Information). The UV−vis titration of RHES with Al3+ shows the appearance of two new bands at 406 and 555 nm (Figure 3d), indicating the interaction of RHES with Al3+. Job’s plot supports a 1:1 stoichiometry (mole

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RESULTS AND DISCUSSION Colorless RHES displays a very weak emission at 397 nm (λEx, 306 nm, Scheme 1, Figure S1, Supporting Information). In the presence of Al3+, Zn2+, Cd2+, and Hg2+, RHES turns pink (Al3+), Scheme 1. Synthetic Protocols of RHES and R1

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Figure 2. Emission (left) and absorption spectra (right) of RHES (the same media and pH mentioned above) in the presence of the common metal ions (λEx = 306 nm).

Figure 3. (a) Emission intensity of RHES in the presence of Al3+ and the other common metal ions (i) λEx = 306 nm, λEm = 573 nm, red bar and (ii) inset λEx = 306 nm, λEm = 362 nm, sky blue bar; (b) changes in the fluorescence spectra of RHES (20 μM) upon the gradual addition of Al3+ (0.005, 0.01, 0.05, 0.1, 0.5, 1.0, 2.0, 5.0, 10, 50, 100, 200, 500, 800, 1200, and 1600 μM) (λEx = 306 nm); (c) plot of emission intensities of RHES vs [Al3+] at 573 nm (red balls) and at 362 nm (sky blue balls, inset), λEx = 306 nm; (d) changes in the absorption spectra of RHES in the said media upon the gradual addition of Al3+ (same as (b)).

shown in Figure S11 (Supporting Information). The fluorescence titration and emission intensities versus [Zn2+] plots are shown in Figure 4b,c. The absence of bands at 573 nm/577 nm, responsible for bare eye pink/red color, indicates the noninvolvement of the rhodamine moiety to coordinate to Zn2+. RHES efficiently detects Zn2+ at the physiological pH of 7.4 (Figure S2, Supporting Information) and forms a 1:1 (mole ratio) complex at a low [Zn2+], whereas a 1:2 (mole ratio) complex is formed at a higher [Zn2+], as revealed from Job’s plot (Figure S12, Supporting Information) and the mass spectrum. The binding constant of 2.11 × 104 M−1 is indicative of a fairly strong interaction between RHES and Zn2+ (Figure S13, Supporting Information). The UV−vis titration of RHES versus

ratio) between RHES and Al3+ (Figure S7, Supporting Information). The mass spectrum of the [RHES−Al3+] adduct also affirms this composition. The binding constant of RHES for Al3+ is 1.33 × 105 (Figure S8, Supporting Information). In the presence of Zn2+, RHES experiences a red shift of the emission band to bright green along with a 29-fold fluorescence enhancement (λEm, 482 nm; λEx = 306 nm; Φ, 5.02-fold enhancement, Figure 4a) without any interference from the other common cations (Figure S9, Supporting Information). With an increasing [Zn2+], the emission intensity at 342 nm decreases in a ratiometric manner with an isoemissive point at 419 nm (Figure S10, Supporting Information). The plot of emission intensities versus [Zn2+] at two different wavelengths is 4264


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Figure 4. (a) Emission intensity of RHES (20 μM) in the presence of Zn2+ and the other common metal ions (λEx = 306 nm, λEm = 482 nm, olive green bar); (b) fluorescence spectra of RHES (20 mM HEPES-buffered MeOH/H2O, 4/1, v/v, pH 7.4) with increasing [Zn2+] (0.0, 0.05, 0.1, 1.0, 2.0, 5.0, 10, 20, 30, 50, 75, 100, 200, 300, 500, 1000, and 1600 μM); (c) plot of emission intensities of RHES with added Zn2+ (0.05−1600 μM) at 482 nm (red balls); (d) changes in the absorption spectra of RHES with increasing [Zn2+] (same as (b)).

Information). RHES can detect Cd2+ as low as 6.27 × 10−9 M (Figure 5c, inset). In the presence of Hg2+, the emission band of RHES undergoes a red shift with an 89-fold fluorescence enhancement (λEm, 577 nm; λEx = 306 nm; Φ, 6.5-fold increase; Figure 6a while its solution color turns intense bloodred under UV light and bare eye (Figure 1). Upon a gradual addition of Hg2+, the emission intensity at 375 nm decreases in a ratiometric manner with an isoemissive point at 526 nm (Figure S19, Supporting Information). The plot of emission intensity versus [Hg2+] at two different wavelengths is shown in Figure S20 (Supporting Information). The fluorescence titration spectra and the plot of emission intensity versus [Hg2+] are shown in Figure 6b,c. The UV−vis titration of RHES versus [Hg2+] is presented in Figure 6d. The optimum pH for the entire study has been determined from Figure S2 (Supporting Information) as 7.4. Figure S21 (Supporting Information) reveals that the common cations do not interfere with the determination of Hg2+. Similar to Al3+, Hg2+ also forms a 1:1 (mole ratio) adduct with RHES, realized from Job’s plot (Figure S22, Supporting Information) and supported by the mass spectrum. The strong interaction between RHES and Hg2+ is indicated from the binding constant, 4.09 × 105 M−1 (Figure S23, Supporting Information). RHES detects as low as 1.7 × 10−10 M Hg2+ (Figure 6c, inset). As the literature suggests, probably the present study is the first report of a single sensor that detects and discriminates Al3+, Zn2+, Cd2+, and Hg2+ through the generation of different colors upon

Zn2+ is presented in Figure 4d. RHES can detect Zn2+ as low as 1.2 × 10−9 M (Figure 4c, inset). In the presence of Cd2+, RHES shows a sky blue fluorescence (λEm = 395 nm, λEx = 306 nm, Figure 5a) without any interference from the common cations (Figure S14, Supporting Information). A gradual addition of Cd2+ enhances the emission intensity at 395 nm, while it decreases at 347 nm in a ratiometric manner with an isoemissive point at 368 nm (Figure S15, Supporting Information). The plots of emission intensities versus [Cd2+] at two different wavelengths are shown in Figure S16 (Supporting Information). The fluorescence titration with Cd2+ (0.05−1600 μM) and concentration versus fluorescence intensity are shown in Figure 5b,c. The absence of the fluorescence bands at 573 nm/577 nm at fluorescence titration and at 555 nm at UV titration of RHES with Cd2+ also confirm that there is no coordination with the rhodamine ring (Figure 5d). At a physiological pH (pH = 7.4), RHES is also efficient to detect Cd2+ (Figure S2, Supporting Information). RHES forms a 1:1 complex with Cd2+ at a low Cd2+ concentration and a 1:2 at a higher concentration, which is confirmed from Job’s plot (Figure S17, Supporting Information) and the mass spectra of the RHES−Zn2+ complex. The blue shift of the emission band is accompanied by a 36-fold fluorescence enhancement for Cd2+ at its emission point (λEm, 395 nm; Φ, 4.3-fold enhancement). The binding constant between RHES and Cd2+ is also high enough to form a stable complex (1.35 × 105 M−1) (Figure S18, Supporting 4265


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Figure 5. (a) Fluorescence intensity of RHES in the presence of Cd2+ and other metal ions (λEx = 306 nm, λEm = 395 nm, blue bar); (b) fluorescence spectra of RHES (20 μM) in 20 mM HEPES-buffered MeOH/H2O (4/1, v/v, pH 7.4) upon the addition of increasing amounts of Cd2+ (0.0, 0.05, 0.1, 0.5, 1.0, 5.0, 10, 20, 50, 75, 100, 200, 300, 500, 1000, 1200, and 1600 μM) (λEx = 306 nm); (c) plot of the emission intensities of RHES (20 μM) as a function of externally added Cd2+ (0.05−1600 μM) at 395 nm (red balls); (d) changes in the absorption spectra of RHES (20 μM) in 20 mM HEPESbuffered MeOH/H2O (4/1, v/v, pH 7.4) upon the gradual addition of Cd2+ (same as (b)).

intensity at 573 nm (Figure S27, Supporting Information). The proposed binding mechanism is further substantiated by a control experiment using a model compound R1 that lacks −OH functionality. In the presence of Al3+/Hg2+, the emission spectra of R1 are significantly different (Figures S28, S29, Supporting Information) while their QTOF−MS spectra support the formation of the adduct (Figures S30, S31, Supporting Information). On the basis of these facts, a probable interaction mechanism of RHES with Al3+, Zn2+, Cd2+, and Hg2+ is shown in Scheme 2. As far as the emission profile is concerned (Figure S32, Supporting Information), Zn2+/Cd2+ do not show any significant interaction with R1, suggesting the necessity and involvement of the phenol −OH for their sensing. For a deeper understanding of the binding mode of RHES and to strengthen the proposed sensing mechanism, 1H NMR titration has been conducted in dimethyl sulfoxide (DMSO)-d6. After the addition of 1 equiv Al3+/Hg2+ to RHES, the alkane protons downfield-shifted from 3.45 to 3.50 ppm, indicating the interaction of the adjacent O donors with the cations. Upon the addition of more Al3+/Hg2+ (2.0 and 3.0 equiv), these protons further downfield-shifted to 3.54 ppm. Moreover, the ring protons upfield-shifted from 7.0 to 6.95 ppm, which indicates the opening of the spirolactam ring. Interestingly, after the addition of 1 equiv Al3+/Hg2+ to RHES, the “p” proton downfield-shifted from 8.6 to 10.2 ppm (Al3+) and 10.3 ppm (Hg2+) because of the close proximity of the rhodamine B unit through the Al3+/Hg2+-induced folding of RHES. A further addition of Al3+/Hg2+ (2.0 and 3.0 equiv) shifts

irradiation of UV light and also observable in bare eye (Figure S24, Supporting Information). Al3+ shifts the emission maxima of RHES from 397 nm (characteristic of PET, from the imine N-to the conjugated 3ethoxysalicylaldehyde moiety) to 573 nm (characteristic of rhodamine B, FRET) via an intermediate CHEF process having a characteristic emission at 362 nm. Al3+ being a hard acid prefers an “O” donor from 3-ethoxysalicylaldehyde. The absorption at 406 nm also indicates the interaction between Al3+ and the “O” donor from 3-ethoxysalicylaldehyde (Figure 3d). Thus, upon excitation at 306 nm, the fluorescence at 362 nm enhances because of the CHEF process which subsequently transfers energy to excite the rhodamine moiety. However, Hg2+ being a soft acid does not prefer the hard “O” donor, and hence the CHEF process is absent. Rather, a significant overlap between 3-ethoxysalicylaldehyde emission (donor) and rhodamine absorption (acceptor) has been observed (Figure S25, Supporting Information). Upon the addition of Hg2+, the emission maximum of free RHES is redshifted from 397 to 577 nm (λEx, 306 nm), indicating an energy transfer from the donor to the acceptor. The slow interaction between RHES and Al3+ allows the intermediate CHEF process visible. With time, the intensity at 573 nm gradually increases, and finally it moves over to 362 nm. Hence, an intense pink fluorescence and bare eye pink coloration are observed (Figure S26, Supporting Information). The pseudofirst-order rate constant for the interaction has been estimated as 0.0413 min−1 by monitoring the changes in the emission 4266


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Figure 6. (a) Emission intensity of RHES (20 μM) in the presence of Hg2+ and other common metal ions (λEx = 306 nm, λEm = 577 nm, red bar); (b) changes in the emission spectra of RHES with increasing [Hg2+] (0.005, 0.01, 0.05, 0.1, 0.5, 1.0, 2.0, 5.0, 10, 50, 100, 200, 500, 800, 1000, 1200, 1400, and 1600 μM); (c) plot of emission intensities of RHES as a function of added Hg2+ (0.005−1600 μM, red balls); (d) changes in the absorption spectra of RHES with increasing [Hg2+] (same concentration as (b), along with some more such as 20, 30, 75, 150, and 250 μM) (same media as mentioned earlier).

interaction of the −OH group with Al3+ (Figures S33, S34, Supporting Information). Similarly, the addition of 1 equiv of Zn2+/Cd2+ to RHES shifts the alkane protons downfield from 3.5 to 3.6 ppm, indicating the interaction of the adjacent O donors to the metal ions. These protons are further downfield-shifted upon the addition of more Zn2+/Cd2+ (2.0 and 3.0 equiv). Moreover, the CHN proton (“p” proton) is downfield-shifted from 8.75 to 10.25 ppm, whereas the “v” proton shifts downfield from 12.0 to 12.37, indicating the involvement of the −OH group to Zn2+/Cd2+ binding (Figures S35, 36, Supporting Information). To further support the sensing mechanism, fluorescence lifetime measurement is carried out. The average lifetime (τ) of RHES is 0.3852 ns, whereas the corresponding values for RHES−Al3+ (λEm = 573 nm) and RHES−Hg2+ (λEm = 577 nm) systems are 1.7497 and 1.7257 ns, respectively. The τ values for the RHES−Zn2+ (λEm = 482 nm) and RHES−Cd2+ (λEm = 395 nm) systems are 1.2791 and 1.5333 ns, respectively (Table S1, Supporting Information). The τ value of [RHES−Al3+] at 362 nm is 0.8399 ns (Table S1, Figure S37, Supporting Information). The fluorescence lifetime decay curves along with data fitting are shown in Figure 7. Interestingly, the higher affinity and association constant of RHES for Cd2+ allows easy replacement of Zn2+ from the [RHES−Zn2+] adduct to form a more stable [RHES−Cd2+] adduct (Scheme 3). Thus, the [RHES−Zn2+] adduct turns out to be a better Cd2+ sensor that functions via the displacement

Scheme 2. Proposed Binding Mechanism

the “p” proton more downfield. The position of the “v” proton of RHES remains unchanged upon the addition of Hg2+, indicating the noninteraction of the −OH group with Hg2+, whereas it shifts downfield from 12.01 to 12.44 for Al3+, confirming the 4267


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Figure 7. Fluorescence lifetime decay of RHES and (a) [RHES−Al3+], (b) [RHES−Zn2+], (c) [RHES−Hg2+], and (d) [RHES−Cd2+] adducts at the corresponding emission wavelengths.

It is worth mentioning that the simultaneous presence of Al3+ and Hg2+ can be visualized (Scheme 4) using KI to mask Hg2+, while KI does not interfere with the emission profile of RHES (Figure S38, Supporting Information). This fact is also supported from the mass spectrum of the resulting mixture (Figure S39, Supporting Information). Figure S40 (Supporting Information) pictorially represents the reversibility of the probe toward Hg2+ detection. After the addition of the mixture of Al3+ and Hg2+ to RHES, the characteristic emission of the [RHES−Hg2+] adduct at 577 nm is observed, attributed to the higher affinity of Hg2+ for RHES. The higher binding constant of [RHES−Hg2+] over [RHES−Al3+] also supports the observation (Figures S8, S23, Supporting Information). Figure S41 (Supporting Information) also indicates the formation of the [RHES−Hg2+] adduct (m/z, 1007.86) when the mixture of Al3+ and Hg2+ is added to RHES. Moreover, after the addition of I− to this system, the emission shifts to 573 nm, which is characteristic of the [RHES−Al3+] adduct. After the addition of I−, it captures Hg2+ while only Al3+ binds to RHES, showing an emission at 573 nm. The I−-assisted reversible interaction between the probe RHES and Hg2+ is also established from the 1H NMR titration (Figure S42, Supporting Information). Upon the addition of 1 equiv Hg2+ to the [RHES− Al3+] adduct, the “V” proton of RHES upfield-shifted from 12.34 to 12.15 ppm, indicating the noninteraction of the −OH group with Hg2+ and the replacement of Al3+ by Hg2+. Upon further addition of I− (1 equiv) to the resulting solution, the “V” proton is downfield-shifted from 12.15 to 12.34 ppm, indicating the interaction of the −OH group of RHES. This is due to the

Scheme 3. Easy Replacement of Zn2+ by Cd2+ from the [RHES−Zn2+] Adduct

approach. The gradual addition of Cd2+ to the [RHES−Zn2+] system results in a blue shift of the 482 nm emission band to 395 nm along with a fluorescence enhancement (Figure 8). 4268


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Figure 8. (a) Changes in the fluorescence spectra of the [RHES−Zn2+] adduct upon the addition of Cd2+; (b) changes in the absorbance spectra of the [RHES−Zn2+] adduct upon the addition of Cd2+.

Scheme 4. Role of the Masking Agent (KI) for Simultaneous Detection of Al3+ and Hg2+

interaction of Al3+ present in the system with the free RHES. The formation of this [RHES−Al3+] adduct is also demonstrated from its Fourier transform infrared (FTIR) spectrum (Figure S43, Supporting Information). Table S2 (Supporting Information) compares the present probe with the available probes in the literature, although it is not truly a comparison because none of the available single probes can detect all the four cations simultaneously as demonstrated in the present report. To obtain the energy-optimized geometries of RHES and its Al3+, Hg2+, Zn2+, and Cd2+ adducts (Figure S44, Supporting Information), density functional theoretical (DFT) calculations57 have been performed. The Gaussian-09 revision C.01 program package is used for all calculations. The geometries in gas phase are fully optimized without any restrictions of symmetry in singlet ground states for RHES and its adducts, viz. [Al(RHES)(NO3)3], [Zn(RHES)-

(CH3CH2OH)2(CH3OH)(NO3)]+, [Cd(RHES)(CH3CH2OH)(CH3COO)], and [Hg(RHES)(NO3)]+, along with the gradient-corrected DFT level with the three-parameter fit of exchange and correlation functional of Becke (B3LYP), which includes the correlation functional of Lee, Yang and Parr (LYP).57b The basis set LanL2DZ along with an effective core potential is employed for Al, Zn, Cd, and Hg atoms following the associated valence double-ζ basis set of Hay and Wadt57c along with the 6-31++G** basis set, chosen for hydrogen, carbon, nitrogen and oxygen. The highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) of RHES and its adducts are shown in Figure S45 (Supporting Information). It is clear that the Al3+, Hg2+, Zn2+, and Cd2+ adducts are stabilized by 0.06227, 0.03886, 0.0721, and 0.07823 eV, respectively. In [RHES−Al3+] and [RHES−Hg2+], the HOMOs are found to be majorly localized on the rhodamine moiety, whereas these are localized on the aldehyde moiety in the 4269


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ACS Omega case of the [RHES−Zn2+] and [RHES−Cd2+] adducts. The corresponding LUMOs are found to be minorly localized on the rhodamine moiety. Time-dependent DFT (TDDFT) calculations have been performed on the excited state based on the optimized ground-state geometry in the Conductor-like Polarizable Continuum Model (CPCM). The electronic transition energies are calculated by the TDDFT method in methanol and gas phase (Tables S3−S7, Supporting Information). The outcomes are in good agreement with our experimental data. The results reveal that the energy gap between HOMO and LUMO of its adducts is reasonably lower than that of the free RHES, leading to a more stable species. Considering the emission wavelengths of Zn2+ and Cd2+ and the displacement of Zn2+ by Cd2+, a binary logic gate has been constructed.58 There are two input signals, viz. input X (Zn2+) and input Y (Cd2+), whereas the output signals correspond to the turn-on emission at 482 nm for Zn2+ and 395 nm for Cd2+. For input, Zn2+/Cd2+ is defined as the “1” state and absence of any as the “0” state. Our system uses the OR gate and the combination of OR and NOT gates along with a switch that describes the situation clearly. When the input is applied through an individual, both ions follow the OR gate, and when both Zn2+/Cd2+ are the inputs, the pathway is changed (Figure 9).

Figure 10. Fluorescence microscopic images of squamous epithelial cells after incubation for 2 h (100× objective lens): (a) RHES (20 μM); (b) RHES (20 μM) + Zn2+ (30 μM); (c) RHES (20 μM) + Cd2+ (30 μM); (d) RHES (20 μM) + Hg2+ (30 μM).

mM ethylenediaminetetraacetic acid solution in a 60 mm plate. The cells are washed with phosphate-buffered saline (PBS) (×2), and after washing with PBS (×3), the remaining compounds are removed, and the cells are incubated with the said ions and observed under a fluorescence microscope equipped with a UV filter at ambient temperature. No fluorescence is observed in the cells (Figure 10, RHES) that are not previously exposed to the metal ions, whereas after the addition of the said ions, the nonfluorescence gradually turns green with time for Zn2+, sky blue for Cd2+, and red for Hg2+. Moreover, RHES can easily permeate through the tested living cells without causing any harm (as the cells remain alive even after 2 h of exposure to RHES). The cells treated only with the said ions have no fluorescence. This intracellular imaging clearly shows that RHES has good cell permeability and is efficient for imaging of the said cations. Application. Real Sample Analysis. To evaluate the practical feasibility of RHES for the determination of Al3+, Zn2+, Cd2+, and Hg2+, analyses of the real samples have been performed.9,58 The results are summarized in Tables 1−4, respectively. Table 1 Table 1. Determination of Al3+ in Real Samples sample drinking water industrial water antacidsuspension

2+

Figure 9. Truth table diagram for simultaneous monitoring of Zn and Cd2+ as inputs, attributed to the enhancement of emission intensity as the output.

Imaging Studies. Imaging System. The imaging system is composed of an inverted fluorescence microscope Leica DM 1000 LED, a digital compact camera Leica DFC 420C, and an image processor Leica Application Suite v3.3.0. The microscope is equipped with a 50 W mercury arc lamp in a 25 min interval after incubation with the said ions to RHES. Staining with RHES. The slides are deparaffinized in xylene, hydrated through a graded series of ethanol, equilibrated in 1 mL Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS) at 37 °C with 5% CO2, incubated with RHES for 2 min, washed several times with 2% DMSO, and observed under a fluorescence microscope. Image Analysis. RHES efficiently images Zn2+, Cd2+, and Hg2+ in squamous epithelial cells under a fluorescence microscope (Figure 10). The cells are cultured in a 35 mm culture plate at a seeding density of 40 × 104 cell/35 mm in 1 mL DMEM with 10% FBS at 37 °C with 5% CO2. To explore the suitability of RHES for the bioimaging of Zn2+, Cd2+, and Hg2+, the cells are incubated in the presence or absence of 20 mg mL−1 of RHES for 2 h at 37 °C and 5% CO2 in a culture medium. Before this, the cells are subcultured using 0.25% trypsin−0.53

Al3+ added (μM)

emission intensity (au)

Al3+ found (μM)

RSD (%)

recovery (%)

10

423

9.89

1.7

98

12

451

10.7

1.8

561

11.71

1.7

97

indicates the excellent recovery of metal ions from both water and the antacid samples, opening a new avenue for the Table 2. Determination of Hg2+ in Sea Fish Samples sample no. S1 S2 S3 S4 S5 S6 S7 S8

4270

fish name Subgenus Thunnus Eleutheronema tetradactylum Loligo duvauceli Setipinna sp. Johnieops vogleri Stolephorus indicus Pampus argenteus Parupeneus indicus

dry weight of the samples

Hg2+ (μg/g) present


RDS (%)

1.285

1.81

97

1.7

1.321 0.3536 1.21 0.571

0.149 0.291 0.11 0.239

43 51 35 56

1.5 1.8 1.7 1.7

0.826

0.381

67

1.6

0.467

0.167

41

1.9

0.326


Article

ACS Omega Table 3. Determination of Zn2+ in Real Samples sample no. drinking water industrial water antacidsuspension

Zn2+ added (μM)


Zn2+ found (μM)

RDS (%)

recovery (%)

13

210

12.89

1.7

99

269

11.32

1.9

240

14.76

1.9

15

and 3. The sample preparation and analysis procedures are presented in the Supporting Information. Using the calibration graph (Figure S47, Supporting Information), the amounts of unknown Al3+ and Zn2+ have been determined.

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CONCLUSION The synthesis, characterization, and application of a very simple fluorescence and colorimetric probe, RHES, is described. RHES is exploited for the detection of nanomolar Al3+, Zn2+, Cd2+, and Hg2+ ions in a ratiometric manner involving the PET−CHEF− FRET processes. The method allows their bare eye visualization at a physiological pH. It detects as low as 1.5 × 10−9 M Al3+, 1.2 × 10−9 M Zn2+, 6.7 × 10−9 M Cd2+, and 1.7 × 10−10 M Hg2+. Furthermore, RHES successfully images the cations that belong to the same group of the modern periodic table, viz. Zn2+, Cd2+, and Hg2+, in living cells. The developed method is useful for the determination of the said ions in real and sea fish samples. RHES, a multi-ion sensor, has been used to construct a binary logic gate. The DFT studies support the experimental findings.

97

Table 4. Determination of Cd2+ in Sea Fish Samples sample no. S1 S2 S3 S4 S5 S6 S7 S8

fish name Subgenus Thunnus Eleutheronema tetradactylum Loligo duvauceli Setipinna sp. Johnieops vogleri Stolephorus indicus Pampus argenteus Parupeneus indicus

dry weight of the samples

Cd2+ (μg/g) present


RDS (%)

1.375

0.381

51

1.7

1.323 0.273 1.216 0.579

0.176 0.094 0.264 0.291

44 21 32 40

1.7 1.8 1.5 1.7

0.827

0.182

28

1.9

0.557

0.161

31

1.7

0.237

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EXPERIMENTAL SECTION Materials and Methods. High-purity HEPES buffer, rhodamine B, 3-ethoxysalicylaldehyde, 4-formylbenzonitrile, and 2-[2-(2-amino-ethoxy)-ethoxy]-ethylamine have been purchased from Sigma-Aldrich (India). Hg(NO3)2·6H2O, Al(NO3)3·6H2O, Zn(NO3)2·6H2O, and Cd(AcO)2·4H2O are purchased from Merck (India). Spectroscopic grade solvents are used. All metal salts used are of either nitrate or acetate form. The other chemicals are of analytical reagent grade and used without further purification. Ultrapure Milli-Q Millipore 18.2 MΩ cm−1 water is used whenever required. A Shimadzu Multi Spec 2450 spectrophotometer is used for recording the UV−vis spectra. The FTIR spectra are recorded on a Shimadzu FTIR (model IRPrestige 21 CE) spectrophotometer. The mass spectra are recorded with a QTOF 60 Micro YA 263 mass spectrometer in ES-positive mode. The 1H NMR and 13C NMR spectra are measured with Bruker ADVANCE 400 (400 MHz) and 300 (75 MHz) spectrometers. Time-resolved fluorescence lifetime measurements are performed with a picosecond-pulsed diode laser-based time-correlated single-photon counting spectrometer (IBH, UK, λex = 380 nm) coupled to an MCP-PMT detector. The data are fitted to multiexponential functions after the deconvolution of the instrument response function by an iterative reconvolution technique using the IBH DAS 6.2 data analysis software. The steady-state emission and excitation spectra are recorded with a Hitachi F-4500 spectrofluorimeter. A digital SYSTRONICS pH meter (model 335) is used for pH measurement. Synthesis of RHE. 2-[2-(2-Amino-ethoxy)-ethoxy]-ethylamine (7.0 mL, 12.42 mmol, ρ = 1.015 g mL−1) is added dropwise to 1.1 g rhodamine B (2.29 mmol) in methanol (30 mL) under stirring condition at room temperature. The mixture is refluxed for 4 days at 60 °C (Scheme 1). The solvent is removed under reduced pressure using a rotary evaporator. Then, HCl (1 mol L−1) is added until the solution becomes clear. The pH of the solution is adjusted to 9−10 using NaOH (1 mol L−1). A red precipitate that appeared is filtered, washed with water, dried under a vacuum, and assigned as RHE, 2-(2-(2-(2aminoethoxy)ethoxy)ethyl)-3′,6′-bis(diethylamino)spiro[isoindoline-1,9′-xanthen]-3-one (yield: 95.8%). Anal. Calcd (%): C, 71.31; H, 7.74; and N, 9.78. Found: C, 72.05; H, 7.92; and N, 9.25. QTOF−MS ES+ (Figure S48, Supporting

determination of Al3+ in pharmaceutical formulations and water samples. The method has also been employed successfully for the determination of Zn2+ in water and commercially available antacid-suspension supplements (Table 3). Tables 2 and 4 indicate the determination of Hg2+ and Cd2+ ions in sea fish samples, respectively. The results indicate excellent recoveries of the said cations from the real samples. Fish Assays. Collection and Storage. The fish samples are collected from the coastal areas of West Bengal (Digha), India, during January, 2016. Eight fishes, namely subgenus Thunnus, Eleutheronema tetradactylum, Loligoduvauceli, Setipinna sp, Johnieopsvogleri, Stolephorusindicus, Pampus argenteus, and Parupeneusindicus are collected from the coastal areas. The body weight and length of the fishes are measured prior to dissection. The muscle sample is collected and stored in sterile polythene bags and kept in the laboratory deep freezer (−20 °C) to prevent deterioration till further analysis. Analysis. The developed method is used to determine Cd2+ and Hg2+ ions quantitatively in sea fish samples. To obtain the samples in the form of dry powder, the fishes are lyophilized using a Virtis freeze mobile 6EL lyophilizer (UGC-DAE CSR, Kolkata). As the accumulated Cd2+ and Hg2+ in the fish samples are in covalent form, they are not suitable for fluorescence measurements. Hence, the fish samples are digested in a minimum volume of concentrated HNO3 to obtain Cd2+ and Hg2+ in salt form. The sample preparation and analysis procedures are presented in the Supporting Information. The concentrations of Cd2+ and Hg2+ are measured by the standard addition method using the linear plot, as in Figure S46 (Supporting Information). The emission intensity is measured at pH 7.4. Recovery Studies of Zn2+ and Al3+. RHES is also used for the determination of Al3+ and Zn2+ in real samples following the standard addition method described above through spiking the known amounts of these two metal ions at different concentration levels. The results are summarized in Tables 1 4271


Article

ACS Omega Information): [M + H]+ = 573.36 (∼100%). FTIR (cm−1) (Figure S49, Supporting Information): ν(N−H, 1° amine) 3289.23; ν(C−H, aromatic) 2967.21, 2869.56; ν(CO, carbonyl, rhodamine spirolactum ring) 1673.81; ν(CC, aromatic) 1515.22; ν(CC) 1456.23; ν(C−N, carbonyl group) 1389.65; ν(C−O, xanthan ring) 1232.10, 1098.78. Synthesis of RHES. The probe, RHES, has been synthesized by refluxing the equimolar mixture of RHE (1.03 g, 1.80 mmol) and 3-ethoxysalicylaldehyde (0.30 g, 1.80 mmol) in methanol for 6 h at 60 °C (Scheme 1). The red gel obtained after the evaporation of the solvent is assigned as RHES, (E)-3′,6′bis(diethylamino)-2-(2-(2-(2-((2-hydroxy-3-(λ 3 -oxidanyl)benzylidene)amino)ethoxy)ethoxy)ethyl)spiro[isoindoline1,9′-xanthen]-3-one (0.60 g, yield: 96%). Anal. Calcd (%): C, 71.64; H, 7.27; and N, 7.77. Found: C, 71.95; H, 7.37; and N, 7.02. QTOF−MS ES+ (Figure S50, Supporting Information): [M + H]+ = 721.93 (∼40%) and [M + Na]+ = 743.91 (∼100%). 1 H NMR (Figure S51, Supporting Information) (400 MHz, CDCl3): δ (ppm) 13.163 (1H, s), 8.290 (1H, s), 7.895 (5H, m, J = 3.2), 7.433 (5H, m, J = 4.8), 7.062−6.719 (2H, m, J = 2.8), and 6.430−6.228 (4H, m, J = 4.4, xanthene moiety). 13C NMR (Figure S52, Supporting Information) (100 MHz, CDCl3): δ (ppm) 168.39, 166.64, 153.8, 153.34, 152.57, 148.91, 148.81, 132.47, 131.06, 129.03, 128.94, 128.05, 123.87, 123.27, 122.85, 117.78, 115.63, 108.17, 105.61, 97.88, 97.97, 77.47, 77.15, 76.84, 70.62, 69.96, 67.95, 64.96, 58.55, 44.46, and 39.40. FTIR (cm−1) (Figure S53, Supporting Information): ν(O−H) 3373.50; ν(C− H, aromatic) 2974.23, 2904.80; ν(CHN, imine bond) 1678.07; ν(CC, stretch) 1618.28; ν(CO, carbonyl) 1512.19; ν(C−N, stretch) 1381.03; and ν(C−O, stretch) 1232.51, 1068.56. The absorption spectrum (MeOH/H2O, 4/ 1, v/v, 20 mM HEPES, pH 7.4, Figure S54, Supporting Information) shows two peaks, viz. λ (ε, M−1 cm−1) 273 nm (6.3 × 104), assigned to the π−π* transition, and 312 nm (5.93 × 102), assigned to the n−π* transition (from the nonbonding terminal N of the xanthane moiety to π* of RHES). RHES emits at 397 nm (λEx, 306 nm, same media, Figure S52, Supporting Information). Synthesis of R1. The model probe R1 has been synthesized by refluxing an equimolar mixture of RHE (0.50 g, 0.87 mmol) and 4-formylbenzonitrile (0.11 g, 0.87 mmol) in methanol for 6 h at 60 °C (Scheme 1). The red gel obtained after evaporation of the solvent is assigned as R1, (E)-4-(((2-(2-(2-(3′,6′-bis(diethylamino)-3-oxospiro[isoindoline-1,9′-xanthen]-2-yl)ethoxy)ethoxy)ethyl)imino)methyl)benzonitrile (0.108 g, yield, 95%). Anal. Calcd (%): C, 73.64; H, 6.91; and N, 10.21. Found: C, 74.05; H, 7.07; and N, 9.97. QTOF−MS ES+ (Figure S55, Supporting Information): [M + H]+ = 685.6366 (∼100%). FTIR (cm−1) (Figure S56, Supporting Information): ν(C−H, aromatic) 2968.45; ν(CN) 2362.80; ν(CC, stretch) 1664.57 and 1610.56; ν(CO, carbonyl) 1512.19; ν(C−N, stretch) 1381.03; ν(C−O, stretch) 1265.30, 1222.87, and 1114.86. Synthesis of the [RHES−Al3+] Adduct. The methanol solution of Al(NO3)3·6H2O (0.3 g, 0.8 mmol) is added dropwise to a magnetically stirred solution of RHES (0.57 g, 0.8 mmol) in methanol at room temperature. Upon slow evaporation of the solvent, an intense pink color solid has been obtained. The yield is 80%. Anal. Calcd (%): C, 55.30; H, 5.61; N, 10.50; and Al, 2.89. Found: C, 55.98; H, 5.50; and N, 11.40. QTOF−MS ES+ (Figure S57, Supporting Information): m/z, 956.83 (∼100%) is assigned to [RHES + Al3+ + 3NO3− + Na]+, indicating a 1:1 (mole ratio) stoichiometry between RHES and Al3+. FTIR (cm−1) (Figure

S58, Supporting Information): ν(C−H, stretch) 2995.29; ν(C− O, carbonyl) 1618.28; ν(N−O) 1390.68; and ν(C−O) 1305.81. UV−vis (Figure S59, Supporting Information): λ (nm) in MeOH/H2O, 4/1, v/v (ε, M−1 cm−1): 406 nm (5.6 × 102), 555 nm (7.5 × 106). The excitation spectrum (λEm, 573 nm) is presented in Figure S59 (Supporting Information). Synthesis of the [RHES−Zn2+] Adduct. The methanol solution of Zn(NO3)2·6H2O (0.3 g, 1.01 mmol) is added dropwise to a magnetically stirred solution of RHES (0.73 g, 1.01 mmol) in methanol at room temperature. Upon the removal of the solvent by slow evaporation, a light yellow solid appears, which is assigned as the [RHES−Zn2+] adduct (yield, 85%). Anal. Calcd (%): C, 59.53; H, 6.95; and N, 7.21. Found: C, 59.12; H, 6.75; and N, 7.21. QTOF−MS ES+ (Figure S60, Supporting Information): m/z, 992.03 is assigned to [RHES + Zn2+ + 2CH3CH2OH + CH3OH + H]+ (∼100%), indicating a 1:1 (mole ratio) stoichiometry between RHES and Zn2+, whereas m/z, 1123.04 is assigned to [RHES + 2Zn2+ + 2CH3CH2OH + 2NO3− + 2H2O + H]+ (∼40%), indicating a 1:2 (mole ratio) stoichiometry between RHES and Zn2+. FTIR (cm−1) (Figure S61, Supporting Information): ν(C−H) 2970.38; ν(CO, carbonyl) 1739.19; ν(CN, imine bond) 1614.42; ν(CC, aromatic) 1377.17; ν(C−N, attached with the carbonyl group) 137.39; and (C−O, stretch) 1217.08. UV−vis (Figure S62, Supporting Information): λ (nm) in MeOH/H2O, 4/1, v/v (ε, M−1 cm−1): 314 nm (5.6 × 102), 273 nm (7.5 × 106). The excitation spectrum (λEm, 482 nm) is presented in Figure S62 (Supporting Information). Synthesis of the [RHES−Cd2+] Adduct. The methanol solution of Cd(NO3)2·4H2O (0.3 g, 0.7 mmol) is added dropwise to a magnetically stirred solution of RHES (0.38 g, 0.7 mmol) in methanol at room temperature. A slow evaporation of the solvent yielded a gray solid, ascribed as the [RHES−Cd2+] adduct (yield, 82%). Anal. Calcd (%): C, 52.36; H, 5.97; and N, 4.20. Found: C, 52.36; H, 5.89; and N, 4.29. QTOF−MS ES+ (Figure S63, Supporting Information): m/z, 1020.15 is assigned to [RHES + Cd2+ + 2CH3COO− + CH3CH2OH + H]+ (∼100%), indicating a 1:1 (mole ratio) stoichiometry between RHES and Cd2+, whereas m/z, 1170.21 is assigned to [RHES + 2Cd2+ + 2CH3COO− + CH3O− + 3H2O + H]+ (∼50%), confirming a 1:2 (mole ratio) stoichiometry between RHES and Cd2+. FTIR (cm−1) (Figure S64, Supporting Information): ν(C−H) 2970.38; ν(CO, carbonyl) 1737.86; and ν(C−O) 1217.08. UV−vis (Figure S18, Supporting Information): λ (nm) in MeOH/H2O, 4/1, v/v (ε, M−1 cm−1): 273 nm (5.6 × 106), 311 nm (7.5 × 102). The excitation spectrum (λEm, 395 nm) is presented in Figure S65 (Supporting Information). Synthesis of the [RHES−Hg2+] Adduct. The methanol solution of Hg(NO3)2·6H2O (0.3 g, 0.7 mmol) is added dropwise to a magnetically stirred solution of RHES (0.49 g, 0.7 mmol) in methanol at room temperature. A slow evaporation of the solvent produced bloodred solid (yield, 91%). Anal. Calcd (%): C, 52.51; H, 5.33; and N, 7.12. Found: C, 52.93; H, 5.12; and N, 7.02. QTOF−MS ES+ (Figure S66, Supporting Information): m/z, 1007.77 is assigned to [RHES + Hg2+ + NO3− + Na]+ (∼100%), indicating a 1:1 (mole ratio) stoichiometry between RHES and Hg2+. FTIR (cm−1) (Figure S67, Supporting Information): ν(C−H) 2980.02; ν(C−O, carbonyl) 1672.28; ν(C−N) 1377.17; and ν(C−O, attached with the carbonyl group) 1303.88, 1072.42. UV−vis (Figure S68, Supporting Information): λ (nm) in MeOH/H2O, 4/1, v/v (ε, M−1 cm−1): 555 nm (5.6 × 106). The excitation spectrum (λEm, 577 nm) is presented in Figure S68 (Supporting Information). 4272


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Sens. 2017, 2, 1821−1830. (c) Li, S.; Zhang, D.; Xie, X.; Ma, S.; Liu, Y.; Xu, Z.; Gao, Y.; Ye, Y. A novel solvent-dependently bifunctional NIR absorptive and fluorescent ratiometric probe for detecting Fe3+/Cu2+ and its application in bioimaging. Sens. Actuators, B 2016, 224, 661−667. (10) (a) Roat-Malone, R. M.; John, W.; Jersey, J. N. Bioinorganic Chemistry: A Short Course; John Wiley & Sons, Inc., 2002; pp 1−3. (b) Kabat, G. C.; Rohan, T. E. Does excess iron play a role in breast carcinogenesis? An unresolved hypothesis. Cancer Causes Control 2007, 18, 1047−1053. (11) Mason, W. T. Fluorescent and Luminescent Probes for Biological Activity, 2nd ed.; Mason, W. T., Ed.; Academic Press: London, 1999; p 175. (12) (a) Kawanishi, Y.; Kikuchi, K.; Takakusa, H.; Mizukami, S.; Urano, Y.; Higuchi, T.; Nagano, T. Design and Synthesis of Intramolecular Resonance-Energy Transfer Probes for Use in Ratiometric Measurements in Aqueous Solution. Angew. Chem., Int. Ed. 2000, 39, 3438−3440. (b) Tang, J.; Ma, S.; Zhang, D.; Liu, Y.; Zhao, Y.; Ye, Y. Highly sensitive and fast responsive ratiometric fluorescent probe for Cu2+ based on a naphthalimide-rhodamine dyad and its application in living cell imaging. Sens. Actuators, B 2016, 236, 109−115. (13) Fan, J.; Hu, M.; Zhan, P.; Peng, X. Energy transfer cassettes based on organic fluorophores: construction and applications in ratiometric sensing. Chem. Soc. Rev. 2013, 42, 29−43. (14) Robinson, G. H. Aluminium. Chem. Eng. News 2003, 81, 54. (15) Scerri, E. R. The Periodic Table: Its Story and Its Significance; Oxford University, 2007; pp 1−329. (16) (a) Exley, C. Aluminium: Lithosphere to Biosphere (and Back) Sixth Keele Meeting on Aluminium. 26th February to 2nd March 2005, Sixth Keele Meeting on Aluminium. J. Inorg. Biochem. 2005, 99, 1747− 1928. (b) Ding, P.; Wang, J.; Cheng, J.; Zhao, Y.; Ye, Y. Three Nstablization rhodaminebased fluorescent probes for Al3+ via Al3+promoted hydrolysis of Schiff base. New J. Chem. 2015, 39, 342−348. (17) Altschuler, E. Aluminum-containing antacids as a cause of idiopathic Parkinson’s disease. Med. Hypotheses 1999, 53, 22−23. (18) Wang, B.; Xing, W.; Zhao, Y.; Deng, X. Effects of chronic aluminum exposure on memory through multiple signal transduction pathways. Environ. Toxicol. Pharmacol. 2010, 29, 308−313. (19) Walton, J. R. Aluminum in hippocampal neurons from humans with Alzheimer’s disease. NeuroToxicology 2006, 27, 385−394. (20) Polizzi, S. Neurotoxic Effects of Aluminium Among Foundry Workers and Alzheimer’s Disease. NeuroToxicology 2002, 23, 761−774. (21) Dhara, A.; Jana, A.; Guchhait, N.; Ghosh, P.; Kar, S. K. Rhodamine-based molecular clips for highly selective recognition of Al3+ ions: synthesis, crystal structure and spectroscopic properties. New J. Chem. 2014, 38, 1627−1634. (22) Sahana, A.; Banerjee, A.; Das, S.; Lohar, S.; karak, D.; Sarkar, B.; Mukhopadhyay, S. K.; Mukherjee, A. K.; Das, D. A naphthalene-based Al3+ selective fluorescent sensor for living cell imaging. Org. Biomol. Chem. 2011, 9, 5523−5529. (23) Delhaize, E.; Ryan, P. R. Aluminum Toxicity and Tolerance in Plants. Plant Physiol. 1995, 107, 315−321. (24) Vallee, B. L.; Galdes, A. The metallobiochemistry of zinc enzymes. Adv. Enzymol. Relat. Areas Mol. Biol. 1984, 56, 283−430. (25) (a) Vallee, B. L. Introduction to metallothionein. Methods Enzymol. 1991, 205, 3−7. (b) Matrix Metalloproteinases and Inhibitors; Birkedal-Hansen, H., Werb, Z., Welgus, H., Van Wart, H., Eds; Gustav Fischer: Stuttgart, Germany, 1992; pp 5−19. (26) Vallee, B. L.; Auld, D. S. Zinc: biological functions and coordination motifs. Acc. Chem. Res. 1993, 26, 543−551. (27) (a) Hambidge, M.; Krebs, N. Zinc, diarrhea, and pneumonia. J. Pediatr. 1999, 135, 661−664. (b) Ghosh, M.; Ghosh, A.; Ta, S.; Matalobos, J. S.; Das, D. ESIPT-Based Nanomolar Zn2+ Sensor for Human Breast Cancer Cell (MCF7) Imaging. ChemistrySelect 2017, 2, 7426−7431. (c) Kumari, B.; Lohar, S.; Ghosh, M.; Ta, S.; Sengupta, A.; Banerjee, P. P.; Chattopadhyay, A.; Das, D. Structurally Characterized Zn2+ Selective Ratiometric Fluorescence Probe in 100% Water for HeLa Cell Imaging: Experimental and Computational Studies. J. Fluoresc. 2016, 26, 87−103. (d) Ghosh, A.; Ta, S.; Ghosh, M.; Karmakar, S.; Banik, A.; Dangar, T. K.; Mukhopadhyay, S. K.; Das, D. Dual mode

ASSOCIATED CONTENT

S Supporting Information *

The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acsomega.8b00266. 1 H NMR, MS spectra, and fluorescence lifetime data of the probe and its adducts with the analytes; experimental details of different binding constants; LOD plots; interference studies; optimized structures of the probe and its adducts with the corresponding analytes; and calculated bond lengths and angles (PDF)

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AUTHOR INFORMATION

Corresponding Author

*E-mail: [email protected]. Phone: +91-342-2533913. Fax: +91-342-2530452 (D.D.). ORCID

Debasis Das: 0000-0003-0474-0842 Notes

The authors declare no competing financial interest.

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ACKNOWLEDGMENTS M.G. and M.M. gratefully acknowledge CSIR and UGC (New Delhi) for the financial support, whereas S.T. acknowledges UGC-DAE-Kolkata for funding. M.B. sincerely acknowledges B.U. for the fellowship. The authors also thank Dr. A. Ghosh, Department of Chemistry, University of Calcutta, Kolkata, for NMR and cell imaging studies.

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REFERENCES

(1) Chatterjee, A.; Santra, M.; Won, N.; Kim, S.; Kim, J. K.; Kim, S. B.; Ahn, K. H. Selective fluorogenic and chromogenic probe for detection of silver ions and silver nanoparticles in aqueous media. J. Am. Chem. Soc. 2009, 131, 2040−2041. (2) Ni, Y.; Wu, J. Far-red and near infrared BODIPY dyes: synthesis and applications for fluorescent pH probes and bio-imaging. Org. Biomol. Chem. 2014, 12, 3774−3791. (3) Ohashi, A.; Ito, H.; Kanai, C.; Imura, H.; Ohashi, K. Cloud point extraction of iron(III) and vanadium(V) using 8-quinolinol derivatives and Triton X-100 and determination of 10(-7)moldm(-3) level iron(III) in riverine water reference by a graphite furnace atomic absorption spectroscopy. Talanta 2005, 65, 525−530. (4) Liang, Z.-Q.; Wang, C.-X.; Yang, J.-X.; Gao, H.-W.; Tian, Y.-P.; Tao, X.-T.; Jiang, M.-H. A highly selective colorimetric chemosensor for detecting the respective amounts of iron (II) and iron (III) ions in water. New J. Chem. 2007, 31, 906−910. (5) Lunvongsa, S.; Oshima, M.; Motomizu, S. Determination of total and dissolved amount of iron in water samples using catalytic spectrophotometric flow injection analysis. Talanta 2006, 68, 969−973. (6) Tesfaldet, Z. O.; van Staden, J. F.; Stefan, R. I. Sequential injection spectrophotometric determination of iron as Fe(II) in multi-vitamin preparations using 1,10-phenanthroline as complexing agent. Talanta 2004, 64, 1189−1195. (7) Gomes, D. M. C.; Segundo, M. A.; Lima, J. L. F. C.; Rangel, A. O. S. S. Spectrophotometric determination of iron and boron in soil extracts using a multi-syringe flow injection system. Talanta 2005, 66, 703−711. (8) Bobrowski, A.; Nowak, K.; Zarębski, J. Application of a bismuth film electrode to the voltammetric determination of trace iron using a Fe(III)−TEA−BrO3− catalytic system. Anal. Bioanal. Chem. 2005, 382, 1691−1697. (9) (a) Nandi, S.; Das, D. Smart Probe for Multianalyte Signaling: Solvent Dependent Selective Recognition of I−. ACS Sens. 2016, 1, 81− 87. (b) Zhang, L.; Huang, X.; Cao, Y.; Xin, Y.; Ding, L. Fluorescent Binary Ensemble based on Pyrene Derivative and SDS Assemblies as a Chemical Tongue for Discriminating Metal Ions and Brand Water. ACS 4273


Article

ACS Omega ratiometric recognition of zinc acetate: nanomolar detection with in vitro tracking of endophytic bacteria in rice root tissue. Dalton Trans. 2016, 45, 599−606. (28) (a) Vallee, B. L.; Auld, D. S. Active-site zinc ligands and activated H2O of zinc enzymes. Proc. Natl. Acad. Sci. U.S.A. 1990, 87, 220−224. (b) Vallee, B. L.; Auld, D. S. Zinc coordination, function, and structure of zinc enzymes and other proteins. Biochemistry 1990, 29, 5647−5659. (c) Vallee, B. L.; Auld, D. S. Cocatalytic zinc motifs in enzyme catalysis. Proc. Natl. Acad. Sci. U.S.A. 1993, 90, 2715−2718. (d) Vallee, B. L.; Auld, D. S. New perspective on zinc biochemistry: Cocatalytic sites in multizinc enzymes. Biochemistry 1993, 32, 6493−6500. (e) Vallee, B. L.; Falchuk, K. H. The biochemical basis of zinc physiology. Physiol. Rev. 1993, 73, 79−118. (29) (a) Frederickson, C. J.; Koh, J.-Y.; Bush, A. I. The neurobiology of zinc in health and disease. Nat. Rev. Neurosci. 2005, 6, 449−462. (b) Que, E. L.; Domaille, D. W.; Chang, C. J. Metals in neurobiology: probing their chemistry and biology with molecular imaging. Chem. Rev. 2008, 108, 1517−1549. (30) Emami, S.; Hosseinimehr, S. J.; Taghdisi, S. M.; Akhlaghpoor, S. Kojic acid and its manganese and zinc complexes as potential radioprotective agents. Bioorg. Med. Chem. Lett. 2007, 17, 45−48. (31) Huang, Q.; Pan, Z.; Wang, P.; Chen, Z.; Zhang, X.; Xu, H. Zinc(II) and copper(II) complexes of β-substituted hydroxylporphyrins as tumor photosensitizers. Bioorg. Med. Chem. Lett. 2006, 16, 3030− 3033. (32) (a) Nakayama, A.; Hiromura, M.; Adachi, Y.; Sakurai, H. Molecular mechanism of antidiabetic zinc−allixin complexes: regulations of glucose utilization and lipid metabolism. J. Biol. Inorg Chem. 2008, 13, 675−684. (b) Sakurai, H.; Yoshikawa, Y.; Yasui, H. Current state for the development of metallopharmaceutics and anti-diabetic metal complexes. Chem. Soc. Rev. 2008, 37, 2383. (33) (a) Chohan, Z. H.; Arif, M.; Sarfraz, M. Metal-based antibacterial and antifungal amino acid derived Schiff bases: their synthesis, characterization and in vitro biological activity. Appl. Organomet. Chem. 2007, 21, 294−302. (b) Katiyar, A.; Singh, V. P. Synthesis, structural studies and bio-activity of Mn(II), Co(II), Ni(II), Cu(II) and Zn(II) complexes with p -amino acetophenone salicyloyl hydrazine. J. Coord. Chem. 2008, 61, 3200−3212. (c) Kaczmarek, M. T.; Jastrząb, R.; Hołderna-Kędzia, E.; Radecka-Paryzek, W. Self-assembled synthesis, characterization and antimicrobial activity of zinc(II) salicylaldimine complexes. Inorg. Chim. Acta 2009, 362, 3127−3133. (d) Anbu, S.; Kamalraj, S.; Varghese, B.; Muthumary, J.; Kandaswamy, M. A Series of Oxyimine-Based Macrocyclic Dinuclear Zinc(II) Complexes Enhances Phosphate Ester Hydrolysis, DNA Binding, DNA Hydrolysis, and Lactate Dehydrogenase Inhibition and Induces Apoptosis. Inorg. Chem. 2012, 51, 5580−5592. (34) Ali, M. M.; Frei, E.; Straub, J.; Breuer, A.; Wiessler, M. Induction of metallothionein by zinc protects from daunorubicin toxicity in rats. Toxicology 2002, 179, 85−93. (35) Dai, Z.; Canary, J. W. Tailoring tripodal ligands for zinc sensing. New J. Chem. 2007, 31, 1708−1718. (36) Jobe, K.; Brennan, C. H.; Motevalli, M.; Goldup, S. M.; Watkinson, M. Modular “click” sensors for zinc and their application in vivo. Chem. Commun. 2011, 47, 6036−6038. (37) Androsov, D. A.; Kishbaugh, T. L. S.; Gribble, G. W. Mn (III)based radical addition reactions of 2-nitroindole with activated CH compounds. Tetrahedron Lett. 2008, 49, 6621−6623. (38) Cuajungco, M. P.; Lees, G. J. Zinc metabolism in the brain: relevance to human neurodegenerative disorders. Neurobiol. Dis. 1997, 4, 137−169. (39) Koh, J.-Y.; Suh, S. W.; Gwag, B. J.; He, Y. Y.; Hsu, C. Y.; Choi, D. W. The role of zinc in selective neuronal death after transient global cerebral ischemia. Science 1996, 272, 1013−1016. (40) Frederickson, C. J.; Hernandez, M. D.; McGinty, J. F. Translocation of zinc may contribute to seizure-induced death of neurons. Brain Res. 1989, 480, 317−321. (41) Küry, S.; Dréno, B.; Bézieau, S.; Giraudet, S.; Kharfi, M.; Kamoun, R.; Moisan, J.-P. Identification of SLC39A4, a gene involved in acrodermatitis enteropathica. Nat. Genet. 2002, 31, 239−240.

(42) Agency for Toxic Substances and Disease Registry, 4770 Buford Hwy NE, Atlanta, GA 30341. (43) United States Environmental Protectioin Agency. http://water. epa.gov/drink. World Health Organization, Avenue Appia 20, 1211 Geneva 27, Switzerland. http://www.who.int/water_sanitation_ health/dwq/chemicals/cadmium/en/.USA; http://www.atsdr.cdc. gov/cercla/07list.html. (44) (a) Friberg, L.; Elinger, C. G.; Kjelstro, T. Cadmium; World Health Organization: Geneva, Switzerland, 1992. (b) Murray, C. B.; Norris, D. J.; Bawendi, M. G. Synthesis and characterization of nearly monodisperse CdE (E = sulfur, selenium, tellurium) semiconductor nanocrystallites. J. Am. Chem. Soc. 1993, 115, 8706−8715. (c) Tan, L.; Chen, Z.; Zhao, Y.; Wei, X.; Li, Y.; Zhang, C.; Wei, X.; Hu, X. Dual channel sensor for detection and discrimination of heavy metal ions based on colorimetric and fluorescence response of the AuNPs DNA conjugates. Biosens. Bioelectron. 2016, 85, 414−421. (45) Nordberg, G. F.; Herber, R. F. M.; Alessio, L. Cadmium in the Human Environment: Toxicity and Carcinogenicity; Oxford University Press: U.K., 1992. (46) (a) Di Natale, F.; Lancia, A.; Molino, A.; Di Natale, M.; Karatza, D.; Musmarra, D. Capture of mercury ions by natural and industrial materials. J. Hazard. Mater. 2006, 132, 220−225. (b) Wang, C.; Zhang, D.; Huang, X.; Ding, P.; Wang, Z.; Zhao, Y.; Ye, Y. A ratiometric fluorescent chemosensor for Hg2+ based on FRET and its application in living cells. Sens. Actuators, B 2014, 198, 33−40. (c) Song, J.; Huai, M.; Wang, C.; Xu, Z.; Zhao, Y.; Ye, Y. A new FRET ratiometric fluorescent chemosensor for Hg2+ and its application in living EC 109 cells. Spectrochim. Acta, Part A 2015, 139, 549−554. (47) www.epa.gov. (b) Boening, D. W. Ecological effects, transport, and fate of mercury: a general review. Chemosphere 2000, 40, 1335− 1351. (c) Gutknecht, J. Inorganic mercury (Hg2+) transport through lipid bilayer membranes. J. Membr. Biol. 1981, 61, 61−66. (48) (a) Sahana, A.; Banerjee, A.; Lohar, S.; Sarkar, B.; Mukhopadhyay, S. K.; Das, D. Rhodamine-Based Fluorescent Probe for Al3+ through Time-Dependent PET−CHEF−FRET Processes and Its Cell Staining Application. Inorg. Chem. 2013, 52, 3627−3633. (b) Sahana, A.; Banerjee, A.; Lohar, S.; Guha, S.; Das, S.; Mukhopadhyay, S. K.; Das, D. Cd(II)-triggered excimer−monomer conversion of a pyrene derivative: time dependent red-shift of monomer emission with cell staining application. Analyst 2012, 137, 3910−3913. (49) Xu, Y.; Xiao, L.; Sun, S.; Pei, Z.; Pei, Y.; Pang, Y. Switchable and selective detection of Zn2+ or Cd2+ in living cells based on 3′-Osubstituted arrangement of benzoxazole-derived fluorescent probes. Chem. Commun. 2014, 50, 7514−7516. (50) Maity, D.; Govindaraju, T. A differentially selective sensor with fluorescence turn-on response to Zn2+ and dual-mode ratiometric response to Al3+ in aqueous media. Chem. Commun. 2012, 48, 1039− 1041. (51) (a) Liu, X.; Zhang, N.; Zhou, J.; Chang, T.; Fang, C.; Shangguan, D. A turn-on fluorescent sensor for zinc and cadmium ions based on perylene tetracarboxylic diimide. Analyst 2013, 138, 901−906. (b) He, H.; Ng, D. K. P. Differential Detection of Zn2+ and Cd2+ Ions by BODIPY-Based Fluorescent Sensors. Chem.Asian J. 2013, 8, 1441− 1446. (c) Chao, D. Highly selective detection of Zn2+ and Cd2+ with a simple amino-terpyridine compound in solution and solid state. J. Chem. Sci. 2016, 128, 133−139. (d) Li, M.; Lu, H.-Y.; Liu, R.-L.; Chen, J.-D.; Chen, C.-F. Turn-On Fluorescent Sensor for Selective Detection of Zn2+, Cd2+, and Hg2+ in Water. J. Org. Chem. 2012, 77, 3670−3673. (52) Szaciłowski, K. Digital Information Processing in Molecular Systems. Chem. Rev. 2008, 108, 3481. (53) Ozlem, S.; Akkaya, E. U. Thinking outside the silicon box: molecular and logic as an additional layer of selectivity in singlet oxygen generation for photodynamic therapy. J. Am. Chem. Soc. 2009, 131, 48− 49. (54) Balzani, V.; Scandola, F. Supramolecular Photochemistry; Horwood: Chichester, U.K., 1991. (55) (a) Yang, Y.-K.; Yook, K.-J.; Tae, J. A rhodamine-based fluorescent and colorimetric chemodosimeter for the rapid detection of Hg2+ ions in aqueous media. J. Am. Chem. Soc. 2005, 127, 16760− 4274


Article

ACS Omega 16761. (b) Yang, H.; Zhou, Z.; Huang, K.; Yu, M.; Li, F.; Yi, T.; Huang, C. Multi-signaling optical-electrochemical sensor for Hg2+ based on a rhodamine derivative with a ferrocene unit. Org. Lett. 2007, 9, 4729− 4732. (c) Lee, M. H.; Wu, J.-S.; Lee, J. W.; Jung, J. H.; Kim, J. S. Highly Sensitive and Selective Chemosensor for Hg2+ Based on the Rhodamine Fluorophore. Org. Lett. 2007, 9, 2501−2504. (d) Shi, W.; Ma, H. Rhodamine B thiolactone: a simple chemosensor for Hg2+ in aqueous media. Chem. Commun. 2008, 1856−1858. (e) Zheng, H.; Qian, Z.-H.; Xu, L.; Yuan, F.-F.; Lan, L.-D.; Xu, J.-G. Switching the Recognition Preference of Rhodamine B Spirolactam by Replacing One Atom: Design of Rhodamine B Thiohydrazide for Recognition of Hg(II) in Aqueous Solution. Org. Lett. 2006, 8, 859−861. (f) Saha, S.; Mahato, P.; Baidya, M.; Ghosh, S. K.; Das, A. An interrupted PET coupled TBET process for the design of a specific receptor for Hg2+ and its intracellular detection in MCF7 cells. Chem. Commun. 2012, 48, 9293−9295. (g) Lu, H.; Qi, S.; Mack, J.; Li, Z.; Lei, J.; Kobayashi, N.; Shen, Z. Facile Hg2+ detection in water using fluorescent self-assembled monolayers of a rhodamine-based turn-on chemodosimeter formed via a “click” reaction. J. Mater. Chem. 2011, 21, 10878−10882. (h) Kwon, J. Y.; Jang, Y. J.; Lee, Y. J.; Kim, K. M.; Seo, M. S.; Nam, W.; Yoon, J. A highly selective fluorescent chemosensor for Pb2+. J. Am. Chem. Soc. 2005, 127, 10107− 10111. (i) Xu, L.; Xu, Y.; Zhu, W.; Sun, X.; Xu, Z.; Qian, X. Modulating the selectivity by switching sensing media: a bifunctional chemosensor selectivity for Cd2+ and Pb2+ in different aqueous solutions. RSC Adv. 2012, 2, 6323−6328. (j) Xiang, Y.; Tong, A.; Jin, P.; Ju, Y. New fluorescent rhodamine hydrazone chemosensor for Cu(II) with high selectivity and sensitivity. Org. Lett. 2006, 8, 2863−2866. (k) Zhang, X.; Shiraishi, Y.; Hirai, T. Cu(II)-Selective Green Fluorescence of a Rhodamine−Diacetic Acid Conjugate. Org. Lett. 2007, 9, 5039−5042. (l) Dujols, V.; Ford, F.; Czarnik, A. W. A Long-Wavelength Fluorescent Chemodosimeter Selective for Cu(II) Ion in Water. J. Am. Chem. Soc. 1997, 119, 7386−7387. (m) Adhikari, S.; Ghosh, A.; Ghosh, M.; Guria, S.; Das, D. Ratiometric sensing of Fe3+ through PET-CHEF-FRET processes: Live cell imaging, speciation and DFT studies. Sens. Actuators, B 2017, 251, 942−950. (56) Huang, J.; Xu, Y.; Qian, X. Rhodamine-based fluorescent off−on sensor for Fe3+ − in aqueous solution and in living cells: 8aminoquinoline receptor and 2 : 1 binding. Dalton Trans. 2014, 43, 5983−5989. (57) (a) Adhikari, S.; Mandal, S.; Ghosh, A.; Das, P.; Das, D. Strategically Modified Rhodamine−Quinoline Conjugate as a CHEFAssisted FRET Probe for Au3+: DFT and Living Cell Imaging Studies. J. Org. Chem. 2015, 80, 8530−8538. (b) Lee, C.; Yang, W.; Parr, R. G. Development of the Colle-Salvetti correlation-energy formula into a functional of the electron density. Phys. Rev. B: Condens. Matter Mater. Phys. 1988, 37, 785−789. (c) Hay, P. J.; Wadt, W. R. Ab initio effective core potentials for molecular calculations. Potentials for K to Au including the outermost core orbitals. J. Chem. Phys. 1985, 82, 299. (d) Hay, P. J.; Wadt, W. R. Ab initio effective core potentials for molecular calculations. Potentials for the transition metal atoms Sc to Hg. J. Chem. Phys. 1985, 82, 270−283. (58) Ghosh, M.; Mandal, S.; Ta, S.; Das, D. Detection and discrimination of Al3+ and Hg2+ using a single probe: nano-level determination, human breast cancer cell (MCF7) imaging, binary logic gate development and sea fish sample analysis. Sens. Actuators, B 2017, 249, 339−347.

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