Decrease In Fluorescence Lifetime Research Articles

Fluorescence lifetime of plant leaves with sub-nanosecond resolution

The study aimed to develop a measurement apparatus for in vivo chlorophyll-a (Chl-a) fluorescence decay measurements of plants by means of time correlated single photon counting. In this approach, sub-nanosecond laser pulses with a repetition rate of 10 MHz are applied to excite the sample, followed by the analysis of arrival times of the emitted fluorescence photons. Photon statistics are generated by iteratively fitting the sum of two exponential functions. The tool was tested on both plastid and in vivo leaf samples of Savoy cabbage (Brassica oleracea var. sabauda) with 3–4 subsequent leaves giving a complete sample coverage starting from the outermost. The Chl-a fluorescence lifetime exhibited a gradual increase in both the isolated plastid suspensions and the in vivo leaf samples towards the innermost leaf layers explained by an increase of natural absence of light (etiolation syndrome). Furthermore, cadmium stress and iron deficiency were investigated on treated sugar beet (Beta vulgaris) samples in vivo using TCSPS measurements. The reduced fluorescence quenching resulted in an increased fluorescence lifetime. Finally, a long-term (10 week) testing of the setup was carried out on Chl-retaining resurrection Haberlea rhodopensis plants protecting themselves by an elevated non-photochemical quenching yielding a decrease of fluorescence lifetime during their desiccation.

Emission Properties of Individual AIE‐Luminogens in Solution at Room Temperature†

Comprehensive SummaryHere, we combined the photon antibunching analysis, fluorescence correlation spectroscopy, and time‐domain fluorescence lifetime imaging microscopy (TD‐FLIM) to study the emission properties of a representative AIE‐luminogen—4,4’‐(benzo[c][1,2,5] thiadiazole‐4,7‐diyl)bis(N,N‐diphenylaniline) (TPA‐BT) at the single emitter level in a tetrahydrofuran (THF)/water solution where water is a non‐solvent for TPA‐BT. Our findings suggest that, at a constant water fraction in the solution, the size of TPA‐BT aggregates increases with the TPA‐BT concentration; TPA‐BT aggregates are not a quantum emitter at room temperature in the solution. Moreover, utilizing TD‐FLIM and a gel trapping technique allowed us to study the fluorescence lifetime of individual TPA‐BT aggregates. Adding a polar solvent like water does not result in an overall decrease in fluorescence lifetime. Rather, it causes the fluorescence lifetime distribution to become wider, and only some molecules experience a decrease in their fluorescence lifetime. These results could represent a step forward in further understanding the photophysics of AIE‐luminogens.

Growth of Cr,Yb:YAG single crystals for self-Q-switched monolithic solid-state lasers

The saturable absorbers play key role in passively Q-switched lasers, and Cr4+-doped crystals present attractive advantages for Yb:YAG lasers. At present the major problem is the low doping concentration of Cr4+ and the effective way to promote Cr3+→Cr4+ conversion. From this view, we introduce divalent additives Ca2+ and Mg2+ as charge compensators to ensure the charge state of Cr4+ in the tervalence environment of Yb:YAG. The results reveal that this method can promote Cr3+→Cr4+ conversion effectively and an optimal CaO/MgO ratio is 1:4. The corresponding Cr0.01at.%,Yb5at.%:YAG single crystals present enhanced absorption coefficients and fluorescence intensity at 941 nm and 1030 nm, and decreased fluorescence lifetimes and decay curves than Yb:YAG. The thermal expansion coefficient and laser damage threshold of Cr,Yb:YAG single crystal are approaching the YAG host material. The results show that Cr,Yb:YAG crystal has good potential for self-Q-switched monolithic solid-state lasers.

Time-Resolved Fluorescence Spectroscopy of Molecularly Imprinted Nanoprobes as an Ultralow Detection Nanosensing Tool for Protein Contaminants.

Currently, optical sensors based on molecularly imprinted polymers (MIPs) have been attracting significant interest. MIP sensing relies on the combination of the MIP's selective capability, which is conveyed to the polymeric material by a template-assisted synthesis, with optical techniques that offer exquisite sensitivity. In this work, we devised an MIP nanoparticle optical sensor for the ultralow detection of serum albumin through time-resolved fluorescence spectroscopy. The Fluo-nanoMIPs (∅~120 nm) were synthetized using fluorescein-O-methacrylate (0.1×, 1×, 10× mol:mol versus template) as an organic fluorescent reporter. The ability of 0.1× and 1×Fluo-nanoMIPs to bind albumin (15 fM-150 nM) was confirmed by fluorescence intensity analyses and isothermal titration calorimetry. The apparent dissociation constant (Kapp) was 30 pM. Conversely, the 10× fluorophore content did not enable monitoring binding. Then, the time-resolved fluorescence spectroscopy of the nanosensors was studied. The 1×Fluo-nanoMIPs showed a decrease in fluorescence lifetime upon binding to albumin (100 fM-150 nM), Kapp = 28 pM, linear dynamic range 3.0-83.5 pM, limit of detection (LOD) 1.26 pM. Selectivity was confirmed testing 1×Fluo-nanoMIPs against competitor proteins. Finally, as a proof of concept, the nanosensors demonstrated detection of the albumin (1.5 nM) spiked in wine samples, suggesting a possible scaling up of the method in monitoring allergens in wines.

Steady-state and fluorescence lifetime quenching of self-assembled diphenylalanine/coumarin nanostructures as a method to determine dissolved O2 in water

A fluorescent system formed by the combination of coumarin derivative and self-assembled peptide nanostructures was used as sensing system to determine dissolved oxygen (O2) in water samples. Detection was based on the sensing system fluorescence quenching by O2, measured by steady-state and time-resolved fluorescence spectroscopy and Stern-Volmer plot to correlate fluorescence intensity to O2 concentration in the samples. Also, fluorescence lifetime decays informed about the mechanisms in which the quenching is occurring, indicating that the sensing system is highly sensitive and selective to O2, since quenching occurs by a non-radiative energy transfer from electronic excited singlet states of the system to the O2 molecule. This process was evidenced by the system fluorescence lifetime decrease, yielding Singlet O2 as indicative that this system can also find other technological applications.

Fluorescence and phosphorescence lifetime imaging reveals a significant cell nuclear viscosity and refractive index changes upon DNA damage

Cytoplasmic viscosity is a crucial parameter in determining rates of diffusion-limited reactions. Changes in viscosity are associated with several diseases, whilst nuclear viscosity determines gene integrity, regulation and expression. Yet how drugs including DNA-damaging agents affect viscosity is unknown. We demonstrate the use of a platinum complex, Pt[L]Cl, that localizes efficiently mostly in the nucleus as a probe for nuclear viscosity. The phosphorescence lifetime of Pt[L]Cl is sensitive to viscosity and provides an excellent tool to investigate the impact of DNA damage. We show using Fluorescence Lifetime Imaging (FLIM) that the lifetime of both green and red fluorescent proteins (FP) are also sensitive to changes in cellular viscosity and refractive index. However, Pt[L]Cl proved to be a more sensitive viscosity probe, by virtue of microsecond phosphorescence lifetime versus nanosecond fluorescence lifetime of FP, hence greater sensitivity to bimolecular reactions. DNA damage was inflicted by either a two-photon excitation, one-photon excitation microbeam and X-rays. DNA damage of live cells causes significant increase in the lifetime of either Pt[L]Cl (HeLa cells, 12.5–14.1 µs) or intracellularly expressed mCherry (HEK293 cells, 1.54–1.67 ns), but a decrease in fluorescence lifetime of GFP from 2.65 to 2.29 ns (in V15B cells). These values represent a viscosity change from 8.59 to 20.56 cP as well as significant changes in the refractive index (RI), according to independent calibration. Interestingly DNA damage localized to a submicron region following a laser microbeam induction showed a whole cell viscosity change, with those in the nucleus being greater than the cytoplasm. We also found evidence of a by-stander effect, whereby adjacent un-irradiated cells also showed nuclear viscosity change. Finally, an increase in viscosity following DNA damage was also observed in bacterial cells with an over-expressed mNeonGreen FP, evidenced by the change in its lifetime from 2.8 to 2.4 ns.

Stability of retinol in liposomes as measured by fluorescence lifetime spectroscopy and FLIM

Retinol shows complex photophysical properties that make it potentially useful as an exogenous or endogenous probe of membrane microenvironment, but it has not been fully explored. In this study, we use bulk fluorescence lifetime measurements and fluorescence lifetime imaging microscopy (FLIM) to examine the stability of retinol in phosphatidylcholine (PC) multilamellar and unilamellar vesicles with and without cholesterol. We find that both light and exposure to ambient temperature and oxygen contribute to retinol degradation, with the addition of an antioxidant such as butylated hydroxytoluene (BHT) essential to provide stability, especially in the absence of cholesterol. With exposure to ultraviolet light to excite its native fluorescence, retinol degrades rapidly and can photosensitize vesicles. Degradation can be measured by a decrease in fluorescence lifetime. In POPC vesicles without cholesterol, BHT leads to initially higher lifetimes compared with no BHT, but it increases the rate of photodegradation. The presence of 10 mol % cholesterol protects against this effect, and vesicles with 20 mol % cholesterol show longer lifetimes without BHT under all conditions. Because of its environmental sensitivity, retinol is interesting as a FLIM probe, but careful controls are needed to avoid degradation, and additional work is needed to optimize liposomes for food and cosmetic applications.

Crystal engineering of Pb(II)-salen coordination polymer enforced for the selective fluorescence NACs sensing activity in a dispersed aqueous medium: A combined experimental and theoretical DFT monologue

A new 1D coordination polymer (CP), [(SCN)Cd(L1)Pb(NCS)]n (1) was synthesized using N2O4-based Scaffold (H2L1 = N, N’-bis(3-methoxysalicylidenimino)-1,3-diaminopropane) and structurally characterized using various analytical instruments. X-ray structure disclosed complex (1) crystallize in the Orthorhombic space group P212121 with Z=4. The crystal structure contains heteronuclear units of [(SCN)Cd(L1)Pb(NCS)]n, linked by a μ1,1,3 thiocyanate (SCN−) bridging. Here weak S···S chalcogen, Pb···S tetrel bonding with two C-H···π(SCN) and one C-H···π(arene) like interactions are vividly observed. The QTAIM/NCI diagram successfully explains the nature of these contacts. Hirshfeld surface (HS) and 2D fingerprint plots analysis has described the non-covalent supramolecular features. The H…H interactions dominate the solid-state crystal structure. The complex fluorescence behaviour is eventually quenched in the presence of explosive nitroaromatic compounds. Complex (1) retained high fluorescence sensitivity and selectivity only for organo-toxin trinitrophenol (TNP). The Stern-Volmer equation evaluated the enhancement of the fluorescence mechanism, the limit of detection (LOD), and the quenching constant (KSV), which for TNP is 6.92 × 104 M−1. The turn-off fluorescence sensing mechanism is explained by (a) AIM/RDG surface interactions energy, (b) Charge-transfer complex, (b) Photo-induced electron transfer (PET) through the HOMO-LUMO approach, (c) Fluorescence resonance energy transfer (FRET), and (d) 1H NMR titration. Notably, the active dynamic sensing is due to a decrease in fluorescence lifetime after the successive addition of TNP with the polymer. Finally, the title complex (1) is employed as a quenched fluorescence response sensor for detecting TNP in a dispersed aqueous medium.

Impact of electromagnetic stimulation on the mechanical and photophysical properties of alfalfa leaves

The aim of the study was to measure the tensile strength of 4-year-old alfalfa leaves cultivated from seeds exposed to pre-sowing stimulation with He–Ne laser light for 1 or 5 min (designated respectively as F1 and F5) and alternating magnetic field with the exposure time of 1 or 5 min (respectively, L1 and L5). The leaves were measured in terms of blade length and width as well as petiole thickness prior to the tensile test. Measurements were also conducted to determine the chlorophyll fluorescence lifetime and content of photosynthetic pigments (chlorophyll a, b, a + b and carotenoids). The observed tensile strength was between 1.59 and 2.45 N. In the test group, the observed strength was lower in leaves collected from the top and central section of the stem but higher in the bottom part of the stem as compared to the control. The maximum increase of the tearing tensile force relative to the control (C) was observed for the L1 and F1 stimulation samples in leaves collected from the top and bottom parts of the stem, while the maximum decrease for that force was recorded for L5 leaves from the middle and top part of the stem. Chlorophyll fluorescence lifetimes and the overall content of photosynthetic pigments (chlorophyll a and b and carotenoids) were noticeably decreased in the leaves subjected to the stressors/stimulants applied. The results obtained for F1, L5 and, L1 stimulation revealed a decrease in fluorescence lifetimes. The content of photosynthetic pigments was also decreased under the influence of laser light stimulation (L1). This was a clear indication of plant ageing.

The Mortality Risk and Pulmonary Fibrosis Investigated by Time-Resolved Fluorescence Spectroscopy from Plasma in COVID-19 Patients.

A method of rapidly pointing out the risk of developing persistent pulmonary fibrosis from a sample of blood is extraordinarily needed for diagnosis, prediction of death, and post-infection prognosis assessment. Collagen scar formation has been found to play an important role in the lung remodeling following SARS-CoV-2 infection. For this reason, the concentration of collagen degradation products in plasma may reflect the process of lung remodeling and determine the extent of fibrosis. According to our previously published results of an in vitro study, an increase in the concentration of type III collagen degradation products in plasma resulted in a decrease in the fluorescence lifetime of plasma at a wavelength of 450 nm. The aim of this study was to use time-resolved fluorescence spectroscopy to assess pulmonary fibrosis, and to find out if the lifetime of plasma fluorescence is shortened in patients with COVID-19. The presented study is thus far the only one to explore the fluorescence lifetime of plasma in patients with COVID-19 and pulmonary fibrosis. The time-resolved spectrometer Life Spec II with the sub-nanosecond pulsed 360 nm EPLED® diode was used in order to measure the fluorescence lifetime of plasma. The survival analysis showed that COVID-19 mortality was associated with a decreased mean fluorescence lifetime of plasma. The AUC of mean fluorescence lifetime in predicting death was 0.853 (95% CI 0.735–0.972, p < 0.001) with a cut-off value of 7 ns, and with 62% sensitivity and 100% specificity. We observed a significant decrease in the mean fluorescence lifetime in COVID-19 non-survivors (p < 0.001), in bacterial pneumonia patients without COVID-19 (p < 0.001), and in patients diagnosed with idiopathic pulmonary fibrosis (p < 0.001), relative to healthy subjects. Furthermore, these results suggest that the development of pulmonary fibrosis may be a real and serious problem in former COVID-19 patients in the future. A reduction in the mean fluorescence lifetime of plasma was observed in many patients 6 months after discharge. On the basis of these data, it can be concluded that a decrease in the mean fluorescence lifetime of plasma at 450 nm may be a risk factor for mortality, and probably also for pulmonary fibrosis in hospitalized COVID-19 patients.

A novel FRET immunosensor for rapid and sensitive detection of dicofol based on bimetallic nanoclusters

To minimize the need of complex testing procedures, accurate, rapid and selective on-site determination of dicofol (DICO) is of vital importance. Herein, fluorescence immunosensing platforms based on the Förster resonance energy transfer (FRET) between Au–Ag bimetallic nanoclusters (Au–Ag NCs) and Au nanoflowers (Au NFs) were developed for the rapid detection of DICO in both liquid system and paper-based analytical devices (PADs). Conjugations of Au–Ag NCs and antigen (NCs-Agen) and Au–Ag NCs were regarded as energy donors in liquid system and PADs, respectively, while conjugations of Au NFs and antibody (NFs-Ab) were synthesized as energy acceptors. The overlapping spectrum and decrease of fluorescence lifetime of NCs-Agen proved that the fluorescence quenching mechanism was FRET effect. When DICO was captured by NFs-Ab, FRET effect was inhibited due to the long distance between NFs-Ab and NCs/NCs-Agen. Under optimal conditions, the FRET-based fluorescent immunosensors allowed for a rapid response toward DICO with considerable sensitivities of 0.185 ng/mL and 0.170 ng/mL in liquid system and PADs, respectively. Excellent specificity and satisfactory recovery in three kinds of tea samples were confirmed as well. This study opens an avenue for on-site, portable and sensitive detection methods.

Effect of halide ions on the fluorescence properties of 3-aminoquinoline in aqueous medium.

Fluorescence (FL) quenching of 3-aminoquinoline (3AQ) by halide ions has been explored in an aqueous acidic medium using the steady-state and time-domain FL measurement techniques. The halide ions showed no significant change in the absorption spectra of 3AQ in an aqueous acidic medium. The FL intensity was strongly quenched by ions and the order of FL quenching by halide ions was . The decrease in FL lifetime along with the reduction in FL intensity of 3AQ suggested the dynamic nature of quenching. The obtained values were 328 for ions and 119 for ions and the values were ~ and , respectively. The observations suggested that the likely governing mechanism for FL quenching may be an electron transfer process and the involvement of the heavy atom effects.

Constructing ultra-stable photothermal plastics assisted by carbon dots with photocaged reactivity

Constructing ultra-stable photothermal plastics assisted by carbon dots with photocaged reactivity

Highly conjugated carbazole and pyrrolo[1,2-a]quinoxaline based small molecules for fluorescent detection of nitroexplosives

Explosive detection has been a need of the current times to ensure security of people & fluorescent small molecules have become popular for their rapid and selective detection. Herein, we report two highly conjugated small organic fluorophores i.e.1,3,6,8-tetraethynyl-9-hexyl-9H-carbazole (Tetra) based on carbazole & 5-(4′-(pyrrolo[1,2-a]quinoxalin-4-yl)-[1,1′-biphenyl]-4-yl)pyrrolo[1,2-c]quinazoline (PQ-1) based on pyrroloquinoxaline scaffolds, both of which showed excellent quenching of fluorescence in the presence of nitroexplosive molecules. Linear Stern Volmer plots were obtained with different quencher molecules giving low detection limits even under visual inspection. A spectral overlap between the absorption spectra of picric acid (PA) and emission spectra of Tetra and a decrease in fluorescence lifetime of fluorophore with increasing concentration of quencher molecules confirm dynamic quenching with Forster Resonance Energy Transfer (FRET) mechanism. PQ-1 on the other hand showed a static mode of quenching indicated by no change in fluorescence lifetimes. DFT studies were done to identify the HOMO-LUMO bandgap, which are in good agreement with the experimental photophysical properties of the molecules. A low detection limit of 6.68 × 10−7 M for picric acid & 2.53 × 10−6 M for 2,4,6-TNT was observed using Tetra. Similarly, detection limit of 4.45 × 10−6 M & 1.84 × 10−6 M was found for PQ-1 towards picric acid & 2,4,6-TNT respectively. Interestingly, both Tetra & PQ-1 sensed aliphatic nitrocompounds like nitromethane with low detection limits of 4.57 × 10−7 M & 9.37 × 10−7 M respectively.

Photodynamic activity of novel cationic porphyrins conjugated to graphene quantum dots against Staphylococcus aureus

Novel 5-(pyridyl)-10-15-20-tris(4-bromophenyl) porphyrin (complex 1), indium metal derivative (complex 2), and quaternized derivative (complex 3) were synthesized and conjugated to graphene quantum dots (GQDs). The conjugation of the porphyrins to GQDs was through [Formula: see text]-[Formula: see text] stacking. Herein, the [Formula: see text]-[Formula: see text] stacking approach was used to avoid covalent conjugation which might compromise the intrinsic chemical and physical properties. The photodynamic activities of the proposed nanomaterials were assessed towards Staphylococcus aureus cell obliteration. The photophysical properties of the prepared complexes were also studied prior to the application. Moreover, a decrease in fluorescence lifetimes was observed upon metalation of complex 1. As anticipated, singlet oxygen quantum yield ([Formula: see text] increased notably upon heavy metal (indium) insertion and upon composite formation. Antimicrobial photodynamic therapy comparative studies were done on quaternized and unquaternized indium porphyrins conjugated to GQDs. Complex 3-GQDs exhibited the highest antibacterial activities compared to other complexes, and this was attributed to the high [Formula: see text] which plays an imperative role in photodynamic therapy applications.

Spatially resolved fluorescence of caesium lead halide perovskite supercrystals reveals quasi-atomic behavior of nanocrystals

We correlate spatially resolved fluorescence (-lifetime) measurements with X-ray nanodiffraction to reveal surface defects in supercrystals of self-assembled cesium lead halide perovskite nanocrystals and study their effect on the fluorescence properties. Upon comparison with density functional modeling, we show that a loss in structural coherence, an increasing atomic misalignment between adjacent nanocrystals, and growing compressive strain near the surface of the supercrystal are responsible for the observed fluorescence blueshift and decreased fluorescence lifetimes. Such surface defect-related optical properties extend the frequently assumed analogy between atoms and nanocrystals as so-called quasi-atoms. Our results emphasize the importance of minimizing strain during the self-assembly of perovskite nanocrystals into supercrystals for lighting application such as superfluorescent emitters.

HydroFlipper membrane tension probes: imaging membrane hydration and mechanical compression simultaneously in living cells.

HydroFlippers are introduced as the first fluorescent membrane tension probes that report simultaneously on membrane compression and hydration. The probe design is centered around a sensing cycle that couples the mechanical planarization of twisted push–pull fluorophores with the dynamic covalent hydration of their exocyclic acceptor. In FLIM images of living cells, tension-induced deplanarization is reported as a decrease in fluorescence lifetime of the dehydrated mechanophore. Membrane hydration is reported as the ratio of the photon counts associated to the hydrated and dehydrated mechanophores in reconvoluted lifetime frequency histograms. Trends for tension-induced decompression and hydration of cellular membranes of interest (MOIs) covering plasma membrane, lysosomes, mitochondria, ER, and Golgi are found not to be the same. Tension-induced changes in mechanical compression are rather independent of the nature of the MOI, while the responsiveness to changes in hydration are highly dependent on the intrinsic order of the MOI. These results confirm the mechanical planarization of push–pull probes in the ground state as most robust mechanism to routinely image membrane tension in living cells, while the availability of simultaneous information on membrane hydration will open new perspectives in mechanobiology.

FLGS-04. Fluorescence lifetime imaging (FLIm) is a dye-free, high sensitivity approach for fluorescence guided surgery in high-grade and low-grade gliomas

Abstract INTRODUCTION Fluorescence-guided surgery can improve tumor identification and extent of surgical resection. 5-ALA is the standard for GBM, but is limited by lack of quantitative fluorescence, a need to work in the dark, and a lack of sensitivity for low grade tumors. We have developed a novel instrument for dye-free tissue autofluorescence lifetime imaging (FLIm) to identify glioma tissue during resection. This approach utilizes time-resolved autofluorescence measurements in narrow-band channels to assess markers of tissue metabolism. Compared to intensity-based imaging of exogenous fluorophores, FLIm has greater sensitivity without dependence on background lighting. The advantages of FLIm include quantitative tissue analysis, the ability to work under full light conditions, sensitivity for high and low grade gliomas, and the potential ability to identify IDH mutational status. In this study, we validated the use of FLIm for identification of glioma tissue at tumor resection margins. METHODS FLIm was used to image tissue margins during glioma resections and compared to microbiopsies from imaged regions to correlate fluorescence with histopathology. RESULTS FLIm was applied intraoperatively to 11 GBM and 5 LGG patients (7 imaged biopsies per patient). In GBM, tumor infiltration of cortex was associated with significantly decreased fluorescence lifetime (FL) in channels 2 (470/28nm;p&amp;lt;0.05) and 3 (542/50nm;p&amp;lt;0.002). In subcortical margins, FL was inversely proportional to the density of tumor in channels 2,3 (p&amp;lt;0.05). When IDH wild-type GBMs were compared to IDH1-mutant tumors, FL was noted to be significantly longer in channel 1 (390/40nm;p&amp;lt;0.05), and trended towards longer FL in channel 2, shorter FL in channel 3. In LGG, FL was significantly correlated with tumor density in channel 2 (p&amp;lt;0.01). CONCLUSIONS FLIm is a dye-free, quantitative alternative to 5-ALA for fluorescence guided glioma resections with sensitivity to high and low-grade tumors, and the ability to predict IDH mutations in GBM. Further validation studies are on-going.

Binding of Calcium and Magnesium Ions to Terrestrial Chromophoric Dissolved Organic Matter (CDOM): A Combination of Steady-State and Time-Resolved Fluorescence Study

Revealing the binding properties of calcium ion (Ca2+) and magnesium ion (Mg2+) to terrestrial chromophoric dissolved organic matter (CDOM) facilities understanding the effect of natural water components on the photophysics of dissolved organic matter. Steady-state and time-resolved fluorescence spectrometry, and dynamic light scattering were applied to investigate the fluorescence quenching process of CDOM by Ca2+ and Mg2+. Due to a remarkable decrease of the steady-state fluorescence intensity and a slight decrease of fluorescence lifetime, the fluorescence quenching of CDOM by cations mainly occurred through a static process. The fluorescence quenching was profound under longer excitation and emission wavelengths. The binding constant (K, L/mol) for Ca2+ to CDOM ranged from 4.29 to 5.09 (lgK), which was approximately one order of magnitude higher than that of Mg2+ to CDOM (3.86 to 4.56). Nevertheless, the efficiency of CDOM fluorescence quenching by Ca2+, Mg2+ was much lower than that by Cu2+. Fluorescence decay became faster in the presence of a high concentration of Ca2+ (&gt;20 mg/L) and Mg2+ (&gt;50 mg/L). In the presence of these two metal ions, particularly for Ca2+, the lifetime of CDOM excited states shifted to the relatively small value side, indicating fluorescence quenching of CDOM mainly occurred through the interaction of Ca2+/Mg2+ with relatively long-lived fluorophores.

Hydrophobic quercetin encapsulated hemoglobin nanoparticles: formulation and spectroscopic characterization

Various natural proteins are finding application in drug delivery for their high biodegradability and biocompatibility. Albumins are well explored and now focus is shifting to other proteins like hemoglobin (Hb) with unique structural properties. In the present study Hb is allowed to denature at pH 5.0 and model hydrophobic drug quercetin (Q) is encapsulated via self-assembly and hydrophobic interactions. Fluorimetric titrations record highest binding between Hb and Q at pH 5.0, rendering significant structural changes in Hb as captured in CD spectra. A decrease in fluorescence life time of tryptophan residues from 3.31 ns in Hb to 2.89 ns in presence of Q at pH 5.0; surmises efficient binding of Q at the hydrophobic core housing tryptophan. Peak shifts in Fourier transform infrared spectroscopy spectra of Hb-Q compared to Hb evidence significant interactions between them at pH 5.0. Significant spectral changes in soret band region of Hb on addition of Q at pH 5.0 envisages unfolding of porphyrin ring and binding influence of Q. Efficient formation of Hb-Q nanoparticles (NPs) at pH 5.0 is established by DLS, SEM and TEM. Communicated by Ramaswamy H. Sarma