Development of a competitive liposome-based lateral flow assay for the rapid detection of the allergenic peanut protein Ara h1

A competitive lateral flow assay for detecting the major peanut allergen, Ara h1, has been developed. The detector reagents are Ara h1-tagged liposomes, and the capture reagents are anti-Ara h1 polyclonal antibodies. Two types of rabbit polyclonal antibodies were raised either against the entire Ara h1 molecules (anti-Ara h1 Ab) or against an immunodominant epitope on Ara h1 (anti-peptide Ab). All of them reacted specifically with Ara h1 in Western Blot against crude peanut proteins. Moreover, the anti-Ara h1 Ab was chosen for this assay development because of its highest immunoactivity to Ara h1-tagged liposomes in the lateral flow assay. The calculated limit of detection (LOD) of this assay is 0.45 g mL^–1 of Ara h1 with a dynamic range between 0.1 and 10 g mL^–1 of Ara h1 in buffer. Additionally, the visually determined detection range is from 1 to 10 g mL^–1 of Ara h1 in buffer. Results using this assay can be obtained within 30 min without the need of sophisticated equipment or techniques; therefore, this lateral flow assay has the potential to be a cost-effective, fast, simple, and sensitive method for on-site screening of peanut allergens.     

Effect of anionic ion-pairing reagent hydrophobicity on selectivity of peptide separations by reversed-phase liquid chromatography

Despite the continuing dominance of trifluoroacetic acid (TFA) as the anionic ion-pairing reagent of choice for peptide separations by reversed-phase high-performance liquid chromatography (RP-HPLC), we believe that a step-by-step approach to re-examining the relative efficacy of TFA compared to other ion-pairing reagents is worthwhile, particularly for the design of separation protocols for complex peptide mixtures, e.g., in proteomics applications. Thus, we applied RP-HPLC in the presence of different concentrations of anionic ion-pairing reagents - phosphoric acid, TFA, pentafluoropropionic acid (PFPA) and heptafluorobutyric acid (HFBA)--to a mixture of three groups of four 10-residue peptides, these groups containing peptides of +1, +3 or +5 net charge. Overall separation of the 12-peptide mixture improved with increasing reagent hydrophobicity (phosphate- < TFA- < PFPA- < HFBA-) and/or concentration of the anion, with reagent hydrophobicity having a considerably more pronounced effect than reagent concentration. HFBA, in particular, achieved an excellent separation at a concentration of just 10 mM, whereby the peptides were separated by charged groups (+1 < +3 < +5) and hydrophobicity within these groups. There was an essentially equal effect of reagent hydrophobicity and concentration on each positive charge of the peptides, a useful observation for prediction of the effect of varying counterion concentration hydrophobicity and/or concentration during optimization of peptide purification protocols. Peak widths were greater for the more highly charged peptides, although these could be decreased significantly by raising the acid concentration; concomitantly, peptide resolution increased with increasing concentration of ion-pairing reagent.     

NMR determination of enantiomeric composition of 1-substituted 3-amino-1,2-dicarba-closo-dodecaboranes using Eu(hfc)3

Experimental conditions for determination of enantiomeric composition of 1-substituted 3-aminocarboranes by ¹H and ¹³C NMR spectroscopy using chiral shift reagent Eu(hfc)₃ have been found.     

Preparation and characterization of lithium ion-conducting glass-ceramics in the Li1 + xCrxGe2 − x(PO4)3 system

Li₂O–Cr₂O₃–GeO₂–P₂O₅ based glasses were synthesized by a conventional melt-quenching method and successfully converted into glass-ceramics through heat treatment. Experimental results of DTA, XRD, ac impedance techniques and FESEM indicated that Li_1.4Cr_0.4Ge_1.6(PO₄)₃ glass-ceramics treated at 900 °C for 12 h in the Li_1 + xCr_xGe_2 − x(PO₄)₃ (x = 0–0.8) system exhibited the best glass stability against crystallization and the highest ambient conductivity value of 6.81 × 10⁻⁴ S/cm with an activation energy as low as 26.9 kJ/mol. In addition, the Li_1.4Cr_0.4Ge_1.6(PO₄)₃ glass-ceramics displayed good chemical stability against lithium metal at room temperature. The good thermal and chemical stability, excellent conducting property, easy preparation and low cost make it promising to be used as solid-state electrolytes for all-solid-state lithium batteries.     

A Novel Fluorescent Aminoacid Designed for Fluorometric Detection of Cu （Ⅱ）

A fluorescent aminoacid was designed for selective and sensitive detection of Cu(II) in aqueous solution. The designing of this Cu(II) fluorescent chemosensing molecule, N ± (1‐naphthyl). aminoacetic acid (NAA), was based on the binding of Cu(II) to aminoacetic acid and the novel charge transfer photophysics of 1‐aminonaphthalenes. The fluorescence of NAA was found quenched by Cu (II) and several other metal ions of similar electronic structure such as Co(II), Ni(II) and Zn(II). The quenching was shown to occur via electron transfer within the metal‐NAA complex, which required an optimal combination of high binding affinity and favorable redox properties of the components in the metal‐NAA complex and hence afforded selective fluorometric detection of Cu(II). The calibration graph obeyed Stern‐Volmer theory and was shown for Cu(II) over the range of 0–2.75 ± 10–⁴ mol/L. The quenching constant of Cu(II) was measured as 8.0 ± 10³ mol/L that was two orders of magnitude higher than those of Co(II), Ni(II) and Zn(II). The 3SD limit of detection for Cu(II) was 8.00 ± 10^?6 mol/L with a coefficient of variation of 1.65%. Linear range for quantitative detection of Cu(II) was 2.67 ± 10^?5‐2.75 ± 10^?4 mol/L. The method was applied to synthetic sample measurements which gave recoveries of 105%‐112%.     

CoCl_2·6H_2O or LaCl_3·7H_2O Catalyzed Biginelli Reaction. One-Pot Synthesis of 3,4-Dihydropyrimidin-2(1 H)-ones

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Enzyme-linked immunosorbent assay for the determination of 20(S)-protopanaxatriol

A highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) method has been developed for the determination of 20(S)-protopanaxatriol (PPT), one of the major aglycones of dammarane-type ginseng saponins. Polyclonal antibodies raised against ginsenoside F₁ (GF₁)-bovine serum albumin showed high reactivities to PPT and GF₁, whereas they exhibited minor or even no cross-reactivities to other ginsenosides and protopanaxadiol (0.19%). The working range of this method extends from 50 pg ml⁻¹ to 20 ng ml⁻¹ of PPT. The assay reported here has been validated against an HPLC technique using PPT-containing samples and was shown to correlate closely (γ=0.993). This ELISA could be a useful tool for the determination of PPT contained in biological fluids and plant materials.     

Pure and doped lanthanum cobaltites obtained by combustion method

We report the synthesis of La_1−xSr_xCoO₃ nanopowders by solution combustion method using metal nitrates and -alanine (alanine method) or urea (urea method) as fuel. The influence of metal nitrates/organic substance molar ratio and the type of fuel was investigated. The isolated complex precursors were characterized by atomic absorption spectroscopy (AAS), FT-IR spectra and DTA–TG analysis. The La_1−xSr_xCoO₃ (x = 0–0.3) powders were characterized by X-ray diffraction (XRD), scanning electron microscopy–energy-dispersive X-ray analysis (SEM–EDX), as well as by specific surface area measurements. XRD patterns indicate the formation of single-phase LaCoO₃ (rhombohedral) when as-synthesized powders were calcined at 873 K, 3 h in the case of the alanine method and at 1073 K, 3 h for urea-based system. Also, strontium doped lanthanum cobaltites obtained by both methods at 1273 K are single phase with rhombohedral perovskite-like structure as XRD data have proved. SEM investigation of pure and doped lanthanum cobaltites reveal that the samples prepared by both methods have fine particles with tendency of agglomerates formation with different shapes, spongy aspect and high porosity. La_1−xSr_xCoO₃ nanopowders obtained by alanine method have larger specific surface area values than those prepared by urea method.     

Highly sensitive catalytic determination of molybdenum

A novel, highly sensitive, selective, and simple kinetic method was developed for the determination of Mo(VI) based on its catalytic effect on the oxidation of 1-amino-2-naphthol-4-sulfonic acid (ANSA) with H₂O₂. The reaction was followed spectrophotometrically by tracing the oxidized product at 465 nm after 30 min of mixing the reagents. The optimum reaction conditions were: 10 mmol l⁻¹ ANSA, 50 mmol l⁻¹ H₂O₂, 100 mmol l⁻¹ acetate buffer of pH 5.0 ± 0.05 and at 40 °C. Addition of 200 μg ml⁻¹ diethylenetriaminepentaacetic acid (DTPA) conferred high selectivity for the proposed method. Following the recommended procedure, Mo(VI) could be determined with a linear calibration graph up to 2.5 ng ml⁻¹ and a detection limit, based on the 3S_b-criterion, of 0.027 ng ml⁻¹. The unique sensitivity and selectivity of the implemented method allowed its direct application to the determination of Mo(VI) in natural and industrial waste water. The method was validated by comparison with the standard ETAAS method. Moreover, published catalytic-spectrophotometric methods for the determination of molybdenum were reviewed.     

Synthesis of 1-oxa-2-silacyclopentane derivatives via intramolecular nucleophilic attack at silicon

Metalation of (HSiMe₂)₃CH with lithium diisopropylamide (LDA) in THF gives (HSiMe₂)₃CLi, which reacts with ethylene oxide, propylene oxide, 1,2-epoxy butane, 1,2-epoxy pentane, 1,2-epoxy hexane, and epichlorohydrin to give the corresponding 1-oxa-2-silacyclopentane derivatives. Then, glycidylmethacrylate (GM) random copolymers with styrene (St) (in a 1:1 and 1:3 mol ratio) were synthesized by solution free radical polymerization at 70(±1) °C using α,α′-azobis(isobutyronitrile) (AIBN) as an initiator. Both types of copolymers were treated with (HSiMe₂)₃CLi to give new modified copolymers. The reaction of (HSiMe₂)₃CLi with epoxides on the side chains of the copolymers does not lead to intramolecular nucleophilic attack contrary to simple epoxides. All the products have been characterized by spectroscopic techniques.