Simple enzyme based fluorimetric biosensor for urea in human biofluids.

As an important biomarker for renal related diseases, detection of urea is playing a vital role in human biofluids on clinical diagnosis concern. In this work, a synthetic salicyaldehyde based imine fluorophore was synthesized using sonication method and conjugated with urease which was used as fluorescent biosensor for the detection of urea in serum samples. This enzyme based biosensor has shown a good selectivity and sensitivity towards urea with the linear range from 2 to 80 mM and the detection limit of 73 µM. The sensing response obtain is highly agreeing with existing analytical technique for urea detection which strongly recommends this biosensor for clinical application.

Optical Immunosensor for the Detection of Listeria monocytogenes in Food Matrixes.

In this paper, simple imine-based organic fluorophore 4-amino-3-(anthracene-9 yl methyleneamino) phenyl (phenyl) methanone (APM) has been synthesized via a greener approach and the same was used to construct a fluorescent immunoassay for the detection of Listeria monocytogenes (LM). A monoclonal antibody of LM was tagged with APM via the conjugation of the amine group in APM and the acid group of anti-LM through EDC/NHS coupling. The designed immunoassay was optimized for the specific detection of LM in the presence of other interfering pathogens based on the aggregation-induced emission mechanism and the formation of aggregates and their morphology was confirmed with the help of scanning electron microscopy. Density functional theory studies were done to further support the sensing mechanism-based changes in the energy level distribution. All photophysical parameters were measured by using fluorescence spectroscopy techniques. Specific and competitive recognition of LM was done in the presence of other relevant pathogens. The immunoassay shows a linear appreciable range from 1.6 × 106-2.7024 × 108 cfu/mL using the standard plate count method. The LOD has been calculated from the linear equation and the value is found as 3.2 cfu/mL, and this is the lowest LOD value reported for the detection of LM so far. The practical applications of the immunoassay were demonstrated in various food samples, and their accuracy obtained was highly comparable with the standard existing ELISA method.

Fluorimetric chemodosimeter for the detection of capsaicinoids in food matrices.

Capsaicin is a major pungent capsaicinoids in chili pepper and it causes duodenal, liver, stomach and gastric cancer in human. Hence, the detection of capsaicinoids becomes important on health issues concern. Here we are reporting, the first organic molecule based fluorimetric sensor for capsaicin detection using simple fluorophore 4-3-(pyren-2-yl-acryloyl) phenyboronic acid (PAPA), which was synthesized via greener microwave method. The probe has detected the capsaicin selectively in presence of other biomolecules in human biofluids through the intramolecular charge transfer mechanism and supported with DFT studies. The sensor has shown an excellent response towards capsaicin from 2 to 40 µM and the limit of detection of 12.84 nM. Real time analysis was done in various food matrices having capsaicinoids and the results have clearly shown good agreement with our optimized data and it also evinced that the developed sensor can be applied to detect the level of pungency of capsaicinoids.

Label free optical biosensor for insulin using naturally existing chromene mimic synthesized receptors: A greener approach.

We currently reporting a simple and first label free optical insulin biosensor using a novel cum naturally existing Chromene mimic receptors via one pot greener methodology. Among synthesized 4H- Chromene derivatives, 2-amino-5,7-dihydroxy-4-(4-nitrophenyl)-4H-chromene-3-carbonitrile (Compound 1c) is showed a very good colorimetric response for insulin within the linear range of 20 fM -500 pM along with theoretically measured LOD of 15.38 fM based on the increment in absorbance. Meanwhile 2-amino-5, 7-dihydroxy-4-(pyren-2-yl)-4H-chromene-3-carbonitrile (Compound 1a) is depicted an excellent quenching in fluorescence response for insulin with LOD of 7.07 fM and along with wide working range from 10 fM -600 pM. The developed biosensor is tested for the detection of insulin in human blood serum using direct as well as standard addition method and the recovery results are showed a very good agreement with standard clinical chemiluminescent assay method. Hence, this developed biosensor is more reliable for clinical detection of Insulin and so a good analytical method for Diabetes diagnosis within a short period of time.

Optical detection of thiocyanate in human saliva based on the colorimetric response of (2-(2-hydroxyphenyl)-1H-benzo[d]imidazol-5-yl)(phenyl) methanone (HBPM)/Co2+ ions conjugate.

Simple colorimetric biosensor was designed for the quantification SCN- ions based on the principle of colorimetric reactions between Co2+ and SCN- ions reaction using synthesized chromophore (2-(2-hydroxyphenyl)-1H-benzo[d]imidazol-5-yl)(phenyl) methanone (HBPM)/Co2+ ions conjugate which was synthesized via greener ultrawave sonication method. The structural characterization of the HBPM chromophore was confirmed by using NMR, ESI-MASS spectral techniques and the photophysical properties, sensor studies were done by using UV-Vis Spectrophotometer. Our designed HBPM/Co2+ ions conjugates have selectively detected SCN- ions qualitatively and quantitatively in the presence of other human salivary interferents. As per clinical point of view, three different ranges of SCN- ions from 0.1 to 2 mM for normal, 3 to 10 mM for disease, and 100-600 µM for below normal ranges were tested successfully by our developed sensor and the LoD was calculated as 5.43 nM. The real potent application of the developed biosensor was tested in human salivary samples of both smokers and non-smokers under different ages and obtained results shown good agreement with existing clinical methods.

Colorimetric detection and bio-magnification of bisphenol A in fish organs and water sources using 3',6'-bis(diethylamino)-2- ((3,4,5trimethyl benzylidene) amino) spiro [isoindoline -1,9'-xanthen ]-3-one (BTSIXO)-Fe3+ ion conjugate.

Current study is focusing mainly on the development of simple, novel, and cost-effective optical sensor to detect and quantify Bisphenol A (BPA) contamination. We designed a very selective and sensitive colorimetric sensor using synthesized 3', 6'- bis(diethylamino) -2- ((3,4,5 trimethyl benzylidene) amino)spiro [isoindoline-1,9'-xanthen] -3-one (BTSIXO) conjugated with Fe3+-ions via very simple eco- friendly synthetic protocol. The sensor has an excellent wide detection range for BPA from 0.1 to 150 ppm with LODs of 0.02 ppm. Finally, the applicability of the sensor was demonstrated in fish samples especially in the organs of Oreochromis mossambicus fingerlings and contaminated industrial water samples. The sensor was also applied for the quantification of BPA present drinking water stored in the plastic bottles. The developed sensor has shown a good agreement and accuracy when compared with ESI-Mass techniques.

Simple and selective optical biosensor using Ultrasonicator synthesis of 5-((anthracen-9-ylmethylene) amino)-2,3-dihydrophthalazine-1,4-dione for direct detection of ascorbic acid in vegetables and fruits.

We report an optical biosensor using imine, 5-((anthrcene-9-ylmethylene) amino)-2,3dihydrophthalazine) 1-4-dione (ADD) for direct detection of ascorbic acid (AA) via FRET quenched. The ADD was successfully prepared by using simple ultra - sonication method, which was characterized by various spectroscopic techniques. The fluorescence intensity of ADD probe was drastically quenched in presence of AA, and shown excellent selectivity towards the detection of AA in presence of possible biological active interferences. A wide linear range from 0.25 to 190 µM was achieved towards the detection of AA with a LOD of 10 nM. The occurrence of FRET mechanism is due to intermolecular hydrogen bonding between ADD and AA, which was confirmed by Density Functional Theory calculations. Moreover, the biosensor was successfully applied for the detection of AA in real samples such as fruits and vegetables to demonstrate the practicability. In addition, the developed biosensor could be a simple and economically cheap platform for the detection of AA in food samples.

Designing of New Optical Immunosensors Based on 2-Amino-4-(anthracen-9-yl)-7-hydroxy-4H-chromene-3-carbonitrile for the Detection of Aeromonas hydrophila in the Organs of Oreochromis mossambicus Fingerlings.

A one-pot greener methodology has been adopted for the synthesis of a simple 4H-chromene core-based fluorescent tag of (S)-2-amino-4-(anthracen-9-yl)-7-hydroxy-4H-chromene-3-carbonitrile (AHC), and its structure has been analyzed using NMR spectroscopy. The physicochemical properties of AHC were well-studied by UV-vis and fluorescent spectroscopy techniques. As a result of excellent emitting property (ϕ ≈ 0.75), it has been coupled with anti-AH through amide linkage, and the AHC-tagged anti-AH has been used as an immunoassay for the selective detection of Aeromonas hydrophila in the presence of interfering pathogens. Under optimized conditions, immunosensors could successfully quantify A. hydrophila from 4 to 736 CFU/mL, and the LOD was 2 CFU/mL. Saliently, the immunoassay has been successfully demonstrated for the analysis of A. hydrophila in the organs of Oreochromis mossambicusfingerlings, and results have shown a very good agreement with our optimized neat AH fluorimetric titration results.

Reply to the Comment on "Michael Addition Based Chemodosimeter for Serum Creatinine Detection Using ( E)-3-(Pyren-2-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-en-1-one Chalcone".

A very simple chemodosimeter has been developed for creatinine biomolecules based on Michael addition reaction by using ( E)-3-(Pyren-2-yl)-1-(3,4,5-trimethoxyphenyl)prop-2- en-1-one Chalcone. The photophysical properties of the chalcone were thoroughly analyzed using UV-vis and emission techniques. The chalcone has exhibited two absorption maxima at 297 and 407 nm which are due n-π* and π-π* transactions, respectively. This property was further confirmed by repeating UV-vis absorbance studies of the chalcones in different solvents having different polarity. The PTP chalcone has originally exhibited ICT mechanism and it is arrested while creatinine is added. However, a ratiometric response is observed due to the creatinine induced ICT mechanism and it is also clearly supported with DFT studies. In our original work, we did DFT studies for only one isomer of the creatinine. Currently, we have extended our DFT studies for another isomer also. The relative quantum yield of the PTP chalcone was calculated in sensing and standard conditions as 0.85 and 0.45, respectively.

Efficient Removal of Cadmium Using Edible Fungus and Its Quantitative Fluorimetric Estimation Using (Z)-2-(4H-1,2,4-Triazol-4-yl)iminomethylphenol.

Microbes accumulate heavy metals after adsorption or absorption. This study exhibited that Trametes versicolor can tolerate up to 5 mg/g concentration of cadmium. Change in fungus morphology due to cadmium along with its absorption were analyzed using SEM, XRD, and EDAX. Cadmium absorption usually increased with time, and it was determined quantitatively by a fluorimetry technique using a synthesized imine fluorophore as a specific probe and compared with results obtained from atomic absorption spectroscopy (AAS). The intensity of the cadmium-specific XRD peak gradually increased up to the seventh day, and the absorption by the organisms reduced the concentration of cadmium even from the effluent of the plating industry. After the seventh day, Trametes versicolor absorbed almost 0.300 mg/g concentration of cadmium as visualized under high content screening from the fluorescence appearance of hyphae. Hence it can be concluded that Trametes versicolor may play a key role in reducing cadmium from a contaminated environment.

Michael Addition Based Chemodosimeter for Serum Creatinine Detection Using ( E)-3-(Pyren-2-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-en-1-one Chalcone.

First, a simple and highly emissive fluorescent chalcone ( E)-3-(pyren-2-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (PTP) was synthesized via simple shaking along with an excellent quantum yield of 0.85, and proved as a stable, highly sensitive, and selective biosensor for creatinine. Owing to its unique photophysical interaction with creatinine through Michael adduct formation, PTP was utilized as a Chemodosimeter for the selective recognition of creatinine in blood serum. Under optimized conditions, a broad range of creatinine detection was achieved  from 0.00000113 mg/dL to 15.8 mg/dL along with an excellent limit of detection of 0.00000065 mg/dL (0.058 nM). This biosensor is highly reproducible even for different concentration levels of creatinine. It is the very first creatinine biosensor possessing a wider linear range for clinical applications for creatinine. To ensure its clinical application, blood serum samples of people of different age groups were collected from Alpha Hospital and analyzed for creatinine by using our chemodosimeter method and compared with data obtained using a commercial method in the Alpha hospital. Our data show very good agreement with clinical data. Because clinical protocol involves trienzymes and tedious sample preparation, no doubt, our chemodosimeter will be a cheap and sensitive option compared to the existing clinical methods.