Phosphate-selective fluorescent sensing microspheres based on uranyl salophene ionophores

Optical dihydrogen phosphate-selective sensors that function on the basis of bulk optode principles and are based on two different uranyl salophene ionophores are reported here for the first time. The influence of the optode composition and measuring conditions such as sample pH on the optode response are characterized, along with sensor selectivity and long-term stability. Three plasticizers of different polarity are considered for optode fabrication: bis(2-ethylhexyl)sebacate (DOS), dodecyl 2-nitrophenyl ether (o-NPDDE), o-nitrophenyloctylether (o-NPOE). The compounds 9-(diethylamino)-5-(octadecanoylimino)-5H-benzo[a]phenoxazine (ETH 5294, chromoionophore I) and 9-(diethylamino)-5-[(2-octyldecyl)imino]benzo[a]phenoxazine (ETH 5350, chromoionophore III) are used as H⁺-selective fluoroionophores that also act as reference ionophores. The resulting optode-based sensors are compared with their ion-selective electrode (ISE) counterparts, and it is revealed that optodes are better suited for operation at physiological pH. The best optode performance was found for the two component optode sensors doped with ETH 5350 and phosphate ionophore(I). The linear range of these sensor was log a = −6.0 to −2.6. Dihydrogen phosphate-selective optode sensors of optimized composition are fabricated in microsphere format and preliminary measurements in diluted sheep blood samples are presented.     

Silane-dextran chemistry on lateral flow polymer chips for immunoassays

The prognosis for patients suffering from cardiovascular and many other diseases can be substantially improved if diagnosed at an early stage. High performance diagnostic testing using disposable microfluidic chips can provide a platform for realizing this vision. Amic AB (Uppsala, Sweden) has developed a new microfluidic test chip for sandwich immunoassays fabricated by injection molding of the cycloolefin-copolymer Zeonor. A highly ordered array of micropillars within the fluidic chip distributes the sample solution by capillary action. Since wetting of the pillar array surface is the only driving force for liquid distribution precise control of the surface chemistry is crucial. In this work we demonstrate a novel protocol for surface hydrophilization and antibody immobilization on cycloolefin-copolymer test chips, based on direct silanisation of the thermoplastic substrate. Dextran is subsequently covalently coupled to amino groups, thus providing a coating with a low contact angle suitable for antibody immobilization. The contact angle of dextran coated chips is stable for at least two months, which enables production of large batches that can be stored for extended periods of time. We demonstrate the utility of the presented platform and surface chemistry in a C-reactive protein assay with a detection limit of 2.6 ng ml(-1), a dynamic range of 10(2) and a coefficient of variance of 15%.     

Tautomerism of uracil probed via infrared spectroscopy of singly hydrated protonated uracil

Tautomerism of the nucleobase uracil is characterized in the gas phase through IR photodissociation spectroscopy of singly hydrated protonated uracil created with tandem mass spectrometric methods in a commercially available Fourier transform ion cyclotron resonance mass spectrometer. Protonated uracil ions generated by electrospray ionization are re-solvated in a low-pressure collision cell filled with a mixture of water vapor seeded in argon. Their structure is investigated by IR photodissociation spectroscopy in the NH and OH stretching region (2500-3800 cm(-1)) with a tabletop IR laser source and in the 1000-2000 cm(-1) range with a free-electron laser. In both regions the IR photodissociation spectrum exhibits well-resolved spectral signatures that point to the presence of two different types of structure for monohydrated protonated uracil, which result from the two lowest-energy tautomers of uracil. Ab initio calculations confirm that no water-catalyzed tautomerization occurs during the re-solvation process, indicating that the two protonated forms of uracil directly originate from the electrospray process.     

Vibrational relaxation pathways of amide I and amide II modes in N-methylacetamide

We studied the vibrational energy relaxation mechanisms of the amide I and amide II modes of N-methylacetamide (NMA) monomers dissolved in bromoform using polarization-resolved femtosecond two-color vibrational spectroscopy. The results show that the excited amide I vibration transfers its excitation energy to the amide II vibration with a time constant of 8.3 ± 1 ps. In addition to this energy exchange process, we observe that the excited amide I and amide II vibrations both relax to a final thermal state. For the amide I mode this latter process dominates the vibrational relaxation of this mode. We find that the vibrational relaxation of the amide I mode depends on frequency which can be well explained from the presence of two subbands with different vibrational lifetimes (~1.1 ps on the low frequency side and ~2.7 ps on the high frequency side) in the amide I absorption spectrum.     

Concentration Dependence of Vitamin C in Combinations with Vitamin E and Zeaxanthin on Light‐Induced Toxicity to Retinal Pigment Epithelial Cells

The purpose of this study was to determine the effects of increasing concentration of ascorbate alone and in combinations with α‐tocopherol and zeaxanthin on phototoxicity to the retinal pigment epithelium. ARPE‐19 cells were exposed to rose bengal and visible light in the presence and absence of antioxidants. Toxicity was quantified by an assay of cell‐reductive activity. A 20 min exposure to visible light and photosensitizer decreased cell viability to ca 42%. Lipophilic antioxidants increased viabilities to ca 70%, 61% and 75% for α‐tocopherol, zeaxanthin and their combination, respectively. Cell viabilities were ca 70%, 56% and 5% after exposures in the presence of 0.35, 0.7 and 1.4 mm ascorbate, respectively. A 45 min exposure increased cell death to ca 74% and >95% in the absence and presence of ascorbate, respectively. In the presence of ascorbate, zeaxanthin did not significantly affect phototoxicity. α‐Tocopherol and its combination with zeaxanthin enhanced protective effects of ascorbate, but did not prevent from ascorbate‐mediated deleterious effects. In conclusion, there is a narrow range of concentrations and exposure times where ascorbate exerts photoprotective effects, exceeding which leads to ascorbate‐mediated increase in photocytotoxicity. Vitamin E and its combination with zeaxanthin can enhance protective effects of ascorbate, but do not ameliorate its deleterious effects.     

Potentiometry at trace levels in confined samples: ion-selective electrodes with subfemtomole detection limits

We explore here for the first time the direct potentiometric detectability of calcium, lead, and silver ions in amounts on the order of 300 attomoles at 100 picomolar concentrations without any preconcentration, analyte recycling, or electrocatalytic signal enhancement. The results presented here place zero-current potentiometry among the most sensitive electrochemical methods available.     

Potentiometric biosensing of proteins with ultrasensitive ion-selective microelectrodes and nanoparticle labels

We report here for the first time on the use of potentiometry for ultrasensitive nanoparticle-based detection of protein interactions. A silver ion-selective microelectrode is used to detect silver ions oxidatively released from silver enlarged gold nanoparticle labels in a sandwich immunoassay. Since potentiometry is expected to largely maintain its analytical characteristics upon reducing the sample volume, it is anticipated that this approach may form the basis for bioassays with attractive detection limits.