Sandra Ast

Lab-in-a-Phone: Smartphone-Based Portable Fluorometer for pH Measurements of Environmental Water

A novel portable fluorometer combining the attributes of a smartphone with an easy-fit, simple and compact sample chamber fabricated using 3-D printing has been developed for pH measurements of environmental water in the field. A color filter attached over the camera white light light-emitting diode selects an excitation band centered around λ ~ 450 nm with a 3-dB bandwidth, Δλ ~ 21 nm. An application-specific, temperature-stable probe based on the 4-aminonaphthalimide fluorophore was synthesized to absorb at this wavelength while emitting in the green region of the visible spectrum. The green emission is readily detected using the smartphone camera and a simple red-green-blue Android application. Suppression of the green emission increases with increasing pH enabling a straightforward pH sensor. The system was calibrated against a commercial spectrofluorometer and pH measurements were taken at various locations around Sydney. The results were then compared directly with those obtained using conventional electrode-based measurements. The data can be stored in the phones available memory or transmitted by phone back to base for further real-time analysis.

A highly K(+)-selective phenylaza-[18]crown-6-lariat-ether-based fluoroionophore and its application in the sensing of K+ ions with an optical sensor film and in cells.

Herein, we report the synthesis of two phenylaza-[18]crown-6 lariat ethers with a coumarin fluorophore (1 and 2) and we reveal that compound 1 is an excellent probe for K(+) ions under simulated physiological conditions. The presence of a 2-methoxyethoxy lariat group at the ortho position of the anilino moiety is crucial to the substantially increased stability of compounds 1 and 2 over their lariat-free phenylaza-[18]crown-6 ether analogues. Probe 1 shows a high K(+)/Na(+) selectivity and a 2.5-fold fluorescence enhancement was observed in the presence of 100 mM K(+) ions. A fluorescent membrane sensor, which was prepared by incorporating probe 1 into a hydrogel, showed a fully reversible response, a response time of 150 s, and a signal change of 7.8% per 1 mM K(+) within the range 1-10 mM K(+). The membrane was easily fabricated (only a single sensing layer on a solid polyester support), yet no leaching was observed. Moreover, compound 1 rapidly permeated into cells, was cytocompatible, and was suitable for the fluorescent imaging of K(+) ions on both the extracellular and intracellular levels.

Integration of the 1,2,3‐Triazole “Click” Motif as a Potent Signalling Element in Metal Ion Responsive Fluorescent Probes

In a systematic approach we synthesized a new series of fluorescent probes incorporating donor-acceptor (D-A) substituted 1,2,3-triazoles as conjugative π-linkers between the alkali metal ion receptor N-phenylaza-[18]crown-6 and different fluorophoric groups with different electron-acceptor properties (4-naphthalimide, meso-phenyl-BODIPY and 9-anthracene) and investigated their performance in organic and aqueous environments (physiological conditions). In the charge-transfer (CT) type probes 1, 2 and 7, the fluorescence is almost completely quenched by intramolecular CT (ICT) processes involving charge-separated states. In the presence of Na(+) and K(+) ICT is interrupted, which resulted in a lighting-up of the fluorescence in acetonitrile. Among the investigated fluoroionophores, compound 7, which contains a 9-anthracenyl moiety as the electron-accepting fluorophore, is the only probe which retains light-up features in water and works as a highly K(+)/Na(+)-selective probe under simulated physiological conditions. Virtually decoupled BODIPY-based 6 and photoinduced electron transfer (PET) type probes 3-5, where the 10-substituted anthracen-9-yl fluorophores are connected to the 1,2,3-triazole through a methylene spacer, show strong ion-induced fluorescence enhancement in acetonitrile, but not under physiological conditions. Electrochemical studies and theoretical calculations were used to assess and support the underlying mechanisms for the new ICT and PET 1,2,3-triazole fluoroionophores.

Incorporating a Piperidinyl Group in the Fluorophore Extends the Fluorescence Lifetime of Click-Derived Cyclam-Naphthalimide Conjugates

Ligands incorporating a tetraazamacrocycle receptor, a ‘click’- derived triazole and a 1,8-naphthalimide fluorophore have proven utility as probes for metal ions. Three new cyclam-based molecular probes are reported, in which a piperidinyl group has been introduced at the 4-position of the naphthalimide fluorophore. These compounds have been synthesized using the copper(I)-catalyzed azide-alkyne Huisgen cycloaddition and their photophysical properties studied in detail. The alkylamino group induces the expected red-shift in absorption and emission spectra relative to the simple naphthalimide derivatives and gives rise to extended fluorescence lifetimes in aqueous buffer. The photophysical properties of these systems are shown to be highly solvent-dependent. Screening the fluorescence responses of the new conjugates to a wide variety of metal ions reveals significant and selective fluorescence quenching in the presence of copper(II), yet no fluorescence enhancement with zinc(II) as observed previously for the simple naphthalimide derivatives. Reasons for this different behaviour are proposed. Cytotoxicity testing shows that these new cyclam-triazole-dye conjugates display little or no toxicity against either DLD-1 colon carcinoma cells or MDA-MB-231 breast carcinoma cells, suggesting a potential role for these and related systems in biological sensing applications.

Synthesis and Evaluation of 1,8-Disubstituted-Cyclam/Naphthalimide Conjugates as Probes for Metal Ions.

Abstract Fluorescent molecular probes for metal ions have a raft of potential applications in chemistry and biomedicine. We report the synthesis and photophysical characterisation of 1,8‐disubstituted‐cyclam/naphthalimide conjugates and their zinc complexes. An efficient synthesis of 1,8‐bis‐(2‐azidoethyl)cyclam has been developed and used to prepare 1,8‐disubstituted triazolyl‐cyclam systems, in which the pendant group is connected to triazole C4. UV/Vis and fluorescence emission spectra, zinc binding experiments, fluorescence quantum yield and lifetime measurements and pH titrations of the resultant bis‐naphthalimide ligand elucidate a complex pattern of photophysical behaviour. Important differences arise from the inclusion of two fluorophores in the one probe and from the variation of triazole substitution pattern (dye at C4 vs. N1). Introducing a second fluorophore greatly extends fluorescence lifetimes, whereas the altered substitution pattern at the cyclam amines exerts a major influence on fluorescence output and metal binding. Crystal structures of two key zinc complexes evidence variations in triazole coordination that mirror the solution‐phase behaviour of these systems.

Bend and twist intramolecular charge transfer and emission for selective metal ion sensing

Zn2+ and Cu2+ complexation of cyclam-triazolyl-naphthalimide fluoro-ionophores lead to increased and decreased fluorescence respectively. The differences between the two metals are accounted for by their Lewis acid and base properties. This difference means the system can be described as an optical diode with characteristics that suggest measurable differences in fluorescence rise and decay times for the mechanical suppression of bend- and/or twist-induced emissions from intramolecular charge transfer. Different fluorescence evolution profiles are observed offering a new way of distinguishing metal ions for applications in biomedical and environmental sensing.

Absorption and fluorescence spectroscopy on a smartphone

A self-powered smartphone-based field-portable “dual” spectrometer has been developed for both absorption and fluorescence measurements. The smartphone’s existing flash LED has sufficient optical irradiance to undertake absorption measurements within a 3D-printed case containing a low cost nano-imprinted polymer diffraction grating. A UV (λex ~ 370 nm) and VIS (λex ~ 450 nm) LED are wired into the circuit of the flash LED to provide an excitation source for fluorescence measurements. Using a customized app on the smartphone, measurements of absorption and fluorescence spectra are demonstrated using pH-sensitive and Zn2+-responsive probes. Detection over a 300 nm span with 0.42 nm/pixel spectral resolution is demonstrated. Despite the low cost and small size of the portable spectrometer, the results compare well with bench top instruments.

Photo- and thermal degradation of olive oil measured using an optical fibre smartphone spectrofluorimeter

Degradation of olive oil under light and heat are analysed using an optical fibre based low-cost portable smartphone spectrofluorimeter. Visible fluorescence bands associated with phenolic acids, vitamins and chlorophyll centred at λ ∼ 452, 525 and 670 nm respectively are generated using near-UV excitation (LED λex ∼ 370 nm), of extra virgin olive oil are degraded more likely than refined olive oil under light and heat exposure. Packaging is shown to be critical when assessing the origin of degradation.

Temperature Controlled Portable Smartphone Fluorimeter

A self-powered temperature-controlled smartphone fluorimeter is demonstrated. The device measures fluorescence for a temperature range of T = 10 to 40 °C with a precision of 0.1 °C. Results can be shared via wireless networking.

Fluorescent measurements of Zn2+ on a smartphone

Using a smartphone-based portable spectrofluorimeter, measurement of metal ion concentration in water is reported. A UV LED (λex ~ 370 nm), which is powered by the internal source of the smartphone was implemented to function as the excitation source. The emission peak of the UV LED overlaps well with the absorption peak of the Zn2+-responsive molecular probe 6-(1,4,8,11-cyclam-1-yl)ethyl-1,2,3-triazol-4-yl)2-ethyl-naphthalimide fluoro-ionophore (λabs ~ 358 nm). The fluorescence emission of this dye at λem ~ 458 nm is enhanced upon coordination of Zn2+. A customized Android application digitally processes the image from a nano-imprinted polymer diffraction grating and analyses the spectral changes. Zn2+ concentration in water samples were measured with a detection limit of δ ~ 5 μM.