Scale-up of high shear granulation based on agitation power

Scale-up of wet granulation in a vertical high shear mixer was conducted. Pharmaceutical excipient mixtures composed of lactose, cornstarch and micro-crystalline cellulose, and hydroxypropylecellulose as a binder were mixed together and then granulated with purified water under various operating conditions and vessel scales. Torque of agitator shaft was continuously measured and then agitation power per unit vessel volume was calculated. The agitation power per unit vessel volume showed a good correlation with physical properties of obtained granules, such as mass median diameter, strength and compressibility. This implied that the scale-up characteristics could be well analyzed by means of the agitation power per unit vessel volume. In addition, the effects of agitator tip speed and Froude number on the agitation power per unit vessel volume were investigated. The results showed that the agitation power per unit vessel volume was well characterized by the tip speed rather than the Froude number. This meant that the granule growth mainly progressed by the shear stress from the agitator blade. Dynamic characteristics of high shear granulation were also discussed here.     

Structures of cleomiscosins,coumarinolignoids of cleome viscosa seeds1

Structures of cleomiscosins A, B and C, three new coumarino-lignoids isolated from the seeds of Cleome viscosa Linné, have been established as 1a, 2a and 1f irrespectively, on the basis of spectral and chemical evidence. The compounds exhibited liver-protective properties.     

Immobilization of Urease and Cholinesterase on the Surface of Semiconductor Transducer for the Development of Light-Addressable Potentiometric Sensors

Various methods for the immobilization of urease and butyrylcholinesterase on the insulator surface of a laser-scanned semiconductor transducer (LSST) have been tested and compared for the development of an enzyme-based light-addressable potentiometric sensor (LAPS). The method of preparing photocurable membranes on LAPS is presented, and a new type of enzyme LAPS with photocurable polymeric enzyme membranes has been elaborated. It was found that sensors prepared by means of covalent bonding and cross-linking with inactive protein (type SIII) and with photocurable membrane matrices (type SIV) are more prospective. The enzyme LAPSensors with photocurable membranes demonstrate a degree of sensitivity close to the theoretical value and working ranges of 6.3·10^–5–1.1·10^–2 and 1·10^–4–1·10^–1molL^–1 urea for acrylamide and acrylate-based membrane matrices, respectively, and 2.5·10^–4–2·10^–1molL^–1 butyrylcholine for an acrylamide membrane matrix. It is shown that such sensors can be also used for the analysis of enzyme inhibitors.     

Analysis of DNA polymorphisms on the human Y-chromosome by microchip electrophoresis

Validation of microchip electrophoresis in DNA analysis has been carried out using an Agilent 2100 Bioanalyzer. With a DNA 500 Assay Kit, the reproducibility and accuracy of fragment sizing of a 10 bp DNA ladder have been shown to be satisfactory with the relative standard deviation and the relative error mostly below 1.0 and 5.0% (n = 12), respectively. Both intraday and interday validations of fragment sizing and quantitation have also been performed with a 7500 Assay Kit (n = 48). Although the results of quantitation are not as good as that of sizing, due to the manual introduction of samples and markers into the chip wells, they are still sufficient to carry out further analyses of practical samples. Based on such reliable results, fast analysis of DNA polymorphisms on the human Y-chromosome has been realized with microchip electrophoresis. The total analysis times of three genomic polymorphisms on the Y-chromosome, Y Alu polymorphism, 47z/StuI, and 12f2, are all within 100 s, and the relative standard deviation and relative error of fragment sizes are below 3.5 and 3.7%, respectively. In addition, a mixture of nine DNA markers on the human Y-chromosome related to examine the cause of spermatogenic failure have been separated successfully with the smallest fragment size difference of 7 bp. Our results demonstrate the potential of microchip electrophoresis in polymorphism analysis with the advantages of high speed, good reproducibility, high precision, and high resolution.     

A new RNA synthetic method with a 2'-O-(2-cyanoethoxymethyl) protecting group

A novel method for the synthesis of RNA oligomers with 2-cyanoethoxymethyl (CEM) as the 2'-hydroxyl protecting group has been developed. The new method allows the synthesis of oligoribonucleotides with an efficiency and final purity comparable to that obtained in DNA synthesis. [structure: see text]     

Effect of polymer matrix and glycerol on rapid single-strand conformation polymorphism analysis by capillary and microchip electrophoresis for detection of mutations in K-ras gene

We present the rapid single-strand conformation polymorphism (SSCP) analysis by capillary and microchip electrophoresis to detect the mutations in K-ras gene. Parameters that might affect the analysis of mutation in K-ras gene, such as the polymer and the additive in the sieving matrix, have been studied systematically. Under the optimal conditions, the analysis of seven mutants of K-ras gene could be finished within 10 min by capillary electrophoresis (CE). Furthermore, with the wild-type gene as the inner standard, the analysis accuracy of mutations could be improved. In addition, by studying the properties of polymer solutions, the matrix suitable for microchip electrophoresis was found, and the detection of mutations in K-ras gene could be further shortened to 1 min.     

Water adsorption on Rh(111) at 20 K: from monomer to bulk amorphous ice

The adsorption of water (D(2)O) molecules on Rh(111) at 20 K was investigated using infrared reflection absorption spectroscopy (IRAS). At the initial stage of adsorption, water molecules exist as monomers on Rh(111). With increasing water coverage, monomers aggregate into dimers, larger clusters (n = 3-6), and two-dimensional (2D) islands. Further exposure of water molecules leads to the formation of three-dimensional (3D) water islands and finally to a bulk amorphous ice layer. Upon heating, the monomer and dimer species thermally migrate on the surface and aggregate to form larger clusters and 2D islands. Based on the temperature dependence of OD stretching peaks, we succeeded in distinguishing water molecules inside 2D islands from those at the edge of 2D islands. From the comparison with the previous vibrational spectra of water clusters on other metal surfaces, we conclude that the number of water molecules at the edge of 2D islands is comparable with that of water molecules inside 2D islands on the Rh(111) surface at 20 K. This indicates that the surface migration of water molecules on Rh(111) is hindered as compared with the cases on Pt(111) and Ni(111) and thus the size of 2D islands on Rh(111) is relatively small.     

A catechol electrode based on spinach leaves

Minced spinach leaf (Spinacea oleracea) has a high activity of catechol oxidase (dimerizing) (EC 1.1.3.14), which is utilized for the determination of catechol by coupling the spinach tissue with a Clark oxygen electrode. The calibration graph for catechol is linear over the range 2×10^?5–8×10^?4 M (RSD 3%). The sensor retains its enzyme activity for at least 18 days. 4-Methylcatechol and glycolate interfere; glucose and ascorbate do not.     

Nonradioactive telomerase activity assay by microchip electrophoresis: privileges to the classical gel electrophoresis assay

The present study accents on the privileges of microchip-based electrophoresis to the conventional gel electrophoresis in separation of telomerase repeat amplification protocol/polymerase chain reaction (PCR) ladder products obtained in telomerase-catalyzed reaction in cancer cells. We try to clarify the interpretation of the results obtained by both electrophoretic procedures and to avoid misinterpretation as a result of PCR-dependent artefacts.     

Vectorial electron relay at ITO electrodes modified with self-assembled monolayers of ferrocene-porphyrin-fullerene triads and porphyrin-fullerene Dyads for molecular photovoltaic devices

Systematic series of indium tin oxide (ITO) electrodes modified covalently with self-assembled monolayers (SAMs) of ferrocene-porphyrin-fullerene triads and porphyrin-fullerene dyads were designed to gain valuable insight into the development of molecular photovoltaic devices. The structures of SAMs on ITO have been investigated by UV/Vis absorption spectroscopy, atomic force microscopy, and cyclic voltammetry. The photoelectrochemical and photophysical (fluorescence lifetime and time-resolved transient absorption) properties were also determined. The highest quantum yield of photocurrent generation (11 %) among donor-acceptor linked systems which are covalently attached to the surface of ITO electrodes was achieved with SAMs of ferrocene-zinc porphyrin-fullerene linked triad on ITO electrodes. The quantum yields of photocurrent generation correlate well with the charge-separation efficiency and the lifetime of the charge-separated state of the porphyrin-fullerene linked systems in solution. These results provide valuable information for the construction of photonic molecular devices and artificial photosynthetic systems on ITO electrodes.