A comparison of liquid chromatography, capillary electrophoresis, and mass spectrometry methods to determine xyloglucan structures in black currants

Different separation (HPAEC, RP-HPLC, CE) and identification (MALDI-TOF-MS, ESI-MS(n)) techniques were compared to analyse oligosaccharides obtained after incubation of xyloglucan with endo-glucanase. It was possible to analyse xyloglucan oligosaccharides with each technique. Several techniques, including off line (HPAEC-MALDI-TOF-MS) or online (CE-ESI-MS(n), RP-HPLC-ESI-MS(n)) connection provided complementary information on xyloglucan structure. Online CE-MS and RP-HPLC-MS are described for the first time in xyloglucan analysis. Advantages and disadvantages of the techniques for different purposes such as structural characterisation of oligosaccharides or oligosaccharide profiling are discussed. Black currant xyloglucans had a rather simple XXXG-type structure with galactose and fucose containing side chains.     

A PARACEST MRI contrast agent to detect enzyme activity

A "smart" PARACEST MRI contrast agent was synthesized to detect caspase-3, which is an important biomarker in apoptosis. The well-known caspase-3 substrate, DEVD (Asp-Glu-Val-Asp), was elongated using the amino group on one sidearm of lanthanide ligand anchored on the polymer support. The amide of DEVD-(Tm-DOTA) showed a PARACEST effect with MR saturation at -51 ppm. DEVD-(Tm-DOTA) amide was successfully cleaved by caspase-3, exposing the free amine group, which showed PARACEST with saturation at +8 ppm. The enzymatic activity of caspase-3 can be detected by the change in PARACEST effect caused by this biotransformation.     

Formation of crystalline TiO2−xNx and its photocatalytic activity

Amorphous precursors to nitrogen-doped TiO₂ (NTP) and pure TiO₂ (ATP) powders were synthesized by hydrolytic synthesis and sol-gel method (SGM), respectively. Corresponding crystalline phases were obtained by thermally induced transformation of these amorphous powders. From FT-IR and XPS data, it was concluded that a complex containing titanium and ammonia was formed in the precipitate stage while calcination drove weakly adsorbed ammonium species off the surface, decomposed ammonia bound on surface of precipitated powder and led to substitution of nitrogen atom into the lattice of TiO₂ during the crystallization. The activation energies required for grain growth in amorphous TiO_2−xN_x and TiO₂ samples were determined to be 1.6 and 1.7 kJ/mol, respectively. Those required for the phase transformation from amorphous to crystalline TiO_2−xN_x and TiO₂ were determined to be 129 and 142 kJ/mol, respectively. A relatively low temperature was required for the phase transformation in NTP sample than in ATP sample. The fabricated N-doped TiO₂ photocatalyst absorbed the visible light showing two absorption edges; one in UV range due to titanium oxide as the main edge and the other due to nitrogen doping as a small shoulder. TiO_2−xN_x photocatalyst demonstrated its photoactivity for photocurrent generation and decomposition of 2-propanol (IPA) under visible light irradiation ().     

Practical method for the synthesis of (R)-homopipecolinic acid and (R)-homoproline esters from ω-chloroalkanoic acids and available chiral amines

A practical synthesis of (R)-homopipecolinic acid methyl ester 1 and (R)-homoproline methyl ester 2 was performed utilizing (i) a direct intramolecular cyclization of ω-chloro-β-enamino esters 11 and 12, which were prepared from available (S)-1-phenylethylamine or (S)-1-(1-naphthyl)ethylamine and ω-chloro-β-keto esters 5 and 10, respectively and (ii) a highly diastereoselective NaBH₄ reduction followed by hydrogenolysis. The present method is a short-step process using inexpensive and readily available substrates and reagents with fewer wasted materials.     

Efficient method for the deprotection of tert-butyldimethylsilyl ethers with TiCl4-Lewis base complexes: application to the synthesis of 1beta-methylcarbapenems

TiCl4-Lewis base (AcOEt, CH3NO2) complexes smoothly deprotected tert-butyldimethylsilyl (TBDMS) ethers. The reaction velocity with these complexes, which seemed less reactive due to the influence of Lewis bases, was considerably greater than that with TiCl4 alone. Selective desilylations between aliphatic and aromatic TBDMS ethers (1 and 5), between 1 and benzyl, allyl, tosyl, methoxyphenyl, and chloroacetyl ethers (13, 14, 15, 16, and 17), and between TBDMS and TBDPS ethers (18 and 19) were successfully performed. Desilylation of TBDMS-aldol, acyloin, and beta-lactam analogues 9-12 proceeded smoothly due to anchimeric assistance by the neighboring carbonyl groups. The present method was successfully applied to the practical synthesis of 1beta-methylcarbapenems 20a'-f'.     

Facile approach for diblock codendrimers by fusion between Fréchet dendrons and PAMAM dendrons

For the first time, a simple and facile approach for the synthesis of diblock codendrimers by fusion between the azide focal point functionalized Fréchet-type polyether and the propargyl focal point functionalized Tomalia-type PAMAM dendrons has been described based on click chemistry, i.e., the copper-catalyzed cycloaddition reaction between alkyne and azide.     

Toward high sequence coverage of proteins in human breast cancer cells using on-line monolith-based HPLC-ESI-TOF MS compared to CE MS

A method is developed toward high sequence coverage of proteins isolated from human breast cancer MCF10 cell lines using a 2-D liquid separations. Monolithic-capillary columns prepared by copolymerizing styrene with divinylbenzene are used to achieve high-resolution separation of peptides from protein digests. This separation is performed with minimal sample preparation directly from the 2-D liquid fractionation of the cell lysate. The monolithic column separation is directly interfaced to ESI-TOF MS to obtain a peptide map. The protein digests were also analyzed by MALDI-TOF MS and an accurate M(r) of the intact protein was obtained using an HPLC-ESI-TOF MS. The result is that these techniques provide complementary information where nearly complete sequence coverage of the protein is obtained and can be compared to the experimental M(r) value. The high sequence coverage provides information on isoforms and other post-translational modifications that would not be available from methods that result in low sequence coverage. The results from the use of monolithic columns are compared to that obtained by CE-MS. The monolithic column separations provide a rugged and highly reproducible method for separating protein digests prior to MS analysis and is suited to confidently identify biomarkers associated with cancer progression.     

A KLM-circuit model of a multi-layer transducer for acoustic bladder volume measurements

In a preceding study a new technique to non-invasively measure the bladder volume on the basis of non-linear wave propagation was validated. It was shown that the harmonic level generated at the posterior bladder wall increases for larger bladder volumes. A dedicated transducer is needed to further verify and implement this approach. This transducer must be capable of both transmission of high-pressure waves at fundamental frequency and reception of up to the third harmonic. For this purpose, a multi-layer transducer was constructed using a single element PZT transducer for transmission and a PVDF top-layer for reception. To determine feasibility of the multi-layer concept for bladder volume measurements, and to ensure optimal performance, an equivalent mathematical model on the basis of KLM-circuit modeling was generated. This model was obtained in two subsequent steps. Firstly, the PZT transducer was modeled without PVDF-layer attached by means of matching the model with the measured electrical input impedance. It was validated using pulse-echo measurements. Secondly, the model was extended with the PVDF-layer. The total model was validated by considering the PVDF-layer as a hydrophone on the PZT transducer surface and comparing the measured and simulated PVDF responses on a wave transmitted by the PZT transducer. The obtained results indicated that a valid model for the multi-layer transducer was constructed. The model showed feasibility of the multi-layer concept for bladder volume measurements. It also allowed for further optimization with respect to electrical matching and transmit waveform. Additionally, the model demonstrated the effect of mechanical loading of the PVDF-layer on the PZT transducer.     

New demodulation filter in digital phase rotation beamforming

In this paper, we present a new quadrature demodulation filter to reduce hardware complexity in digital phase rotation beamforming. Due to its low sensitivity to phase delay errors, digital quadrature demodulation is commonly used in ultrasound machines. However, since it requires two lowpass filters for each channel to remove harmonics, the direct use of conventional finite impulse response (FIR) filters in ultrasound machines is computationally expensive and burdensome. In our new method, an efficient multi-stage uniform coefficient (MSUC) filter is utilized to remove harmonic components in phase rotation beamforming. In comparison with the directly implemented FIR (DI-FIR) and the previously-proposed signed-power-of-two FIR (SPOT-FIR) lowpass filters, the proposed MSUC filter reduces the necessary hardware resources by 93.9% and 83.9%, respectively. In simulation, the MSUC filter shows a negligible degradation in image quality. The proposed method resulted in comparable spatial and contrast resolution to the DI-FIR approach in the phantom study. These preliminary results indicate that the proposed quadrature demodulation filtering method could significantly reduce the hardware complexity in phase rotation beamforming while maintaining comparable image quality.     

Simultaneous Determination of Acetylsalicylic Acid and Caffeine in Pharmaceutical Formulation by First Derivative Synchronous Fluorimetric Method

A sensitive, rapid, and specific assay has been developed for the simultaneous determination of acetylsalicylic acid and caffeine in commercial tablets based on their natural fluorescence. The mixture of these drugs was resolved by first derivative synchronous fluorimetric technique using two scans. At Δλ=106 nm, using first derivative synchronous scanning, only acetylsalicylic acid yields a detectable signal at 316 nm (peak to zero method) which is unaffected by caffeine. At Δλ=30 nm, the signal of caffeine at 288 nm (peak to zero method) is not affected by acetylsalicylic acid. The range of application is between 0.021 and 41.62 μg ml⁻¹ (correlation coefficient, R=0.9995) for acetylsalicylic acid and between 0.4486 and 44.86 μg ml⁻¹ (correlation coefficient, R=0.99786) for caffeine. The recovery range of 98.40–102% for acetylsalicylic acid and 90–100.5% for caffeine from their synthetic mixture was reported. Overall recovery of both compounds about 97–99% for acetylsalicylic acid and 97–98% for caffeine was obtained from real sample analysis. The detection limits are 0.0013 μg ml⁻¹ and 0.0306 μg ml⁻¹ for acetylsalicylic acid and caffeine, respectively. The relative standard deviation (n=10) for 20 μg ml⁻¹ of acetylsalicylic acid is 2.75% and for 2.2 μg ml⁻¹of caffeine is 1.7%.