Flow-injection determination of 3-hdroxybutyrate in serum with an immobilized 3-hydroxybutyrate dehydrogenase reactor and chemiluminescence detection

A flow-injection system for the determination of 3-hydroxybutyrate in serum is described. 3-Hydroxybutyrate dehydrogenase is immobilized on poly(vinyl alcohol) beads and incorporated in a flow-injection system. 1-Methoxy-5-methylphenazinium methylsulphate reacts with enzymatically generated NADH to give H₂O₂, which is detected chemiluminometrically with the reaction of luminol and hexacynoferrate(III). Serum is diluted and filtered through an ultrafiltration membrane. The system responds linearly to injected samples (80 μl) in the concentration range 0.5–300 μM; the detection limit is 0.1 μM. The within-day relative standard deviation (n = 90) for 58 μM 3-hydroxybutyrate in serum is 0.8%. The maximum throughout is 20 samples per hour. The immobilized enzyme is stable for at least 1 month.     

Flow-injection determination of 9,10-phenanthrenequinone with catalytic photometric detection

A catalytic photometric method with a flow-injection system is described for the determination of 9,10-phenanthrenequinone. It is based on the catalytic effect of 9,10-phenanthrenequinone on the redox reaction of 1,2-dinitrobenzene with formaldehyde under alkaline conditions. 9,10-Phenanthrenequinone at the 5.0 x 10(-8)-5.0 x 10(-6)M level can be determined at a rate of 20 samples/hr. The detection limit is 1.0 x 10(-8)M (40 pg in a 10-microl injection).     

Development of a high-performance liquid chromatographic system with enzyme reactors for the determination of N-acetyl branched-chain amino acids

Immobilized enzymes were used as column reactors in a high-performance liquid chromatographic system for the specific detection of N-acetyl branched-chain amino acids (AcBCAs) such as N- acetyl- l -valine (AcVal), N- acetyl- l -leucine (AcLeu) and N- acetyl- l -isoleucine (AcIle). Aminoacylase and leucine dehydrogenase were immobilized onto poly(vinyl alcohol) beads. The AcBCAs were separated as three peaks on a Capcell C(1) SG120 column with 0.03M phosphate buffer (pH 8.0). Aminoacylase was capable of hydrolysing the AcBCAs to amino acids, which react with beta-nicotinamide adenine dinucleotide (NAD(+)) in the presence of leucine dehydrogenase. The reduced nicotinamide adenine dinucleotide (NADH) produced was monitored fluorimetrically. The calibration graphs were linear from 4 to 200muM for AcVal and AcLeu, and from 5 to 300muM for AcIle; detection limits for AcVal, AcLeu and AcIle were 2, 2 and 3muM, respectively. The immobilized aminoacylase reactor should be renewed every 5 days owing to a poor stability of aminoacylase.     

Stereostructures of several solidagolactones (elongatolides)

The structures of several clerodane-type diterpenes [solidagolactones II (elongatolide C), III, IV (elongatolide A), V, VI (elongatolide D), elongatolide B, and elongatolide E] have been revised , all of which are $\text{cis}$-clerodanes.     

A theoretical study of the dielectric solvent effect on vicinal spin coupling constants

The dielectric solvent effect on HCCH, HCNH and HCNC spin coupling constants in ethane, tetrachloroethane and trans N-methylformamide has been calculated by finite perturbation theory based on the INDO and CNDO/2 approximations incorporating solvaton theory. The available experimental data are interpreted using the calculated variations of spin coupling constants. The effect of dielectric constant on the general form of the Karplus relation is included in the finite perturbation calculations.     

Determination of aliphatic amino acids in serum by HPLC with fluorimetric detection using co-immobilized enzyme reactor

A simple and selective method is presented for the determination of aliphatic amino acids such as L-alanine, L-valine, L-isoleucine and L-leucine in serum using HPLC with detection by co-immobilized alanine dehydrogenase/leucine dehydrogenase post-column reactor and fluorimeter. The enzymes were simultaneously immobilized on chitosan beads. The separation was achieved by means of an ods column with elution with phosphate buffer (pH 7.0). The system gave linear responses over two orders of magnitude and detection limits at 1-2muM levels.     

Flow-injection determination of d-mannitol with immobilized mannitol dehydrogenase

A flow-injection system is described for the determination of d-mannitol. Mannitol dehydrogenase is immobilized on poly(vinyl alcohol) beads and packed in a column (5 cm × 4 mm i.d.). The NADH formed is detected fluorimetrically. The response is linear between 5 × 10^?7 and 1 × 10^?4 M mannitol and the detection limit is 1 × 10^?7 M. The throughput is 30 samples per hour. The reactor is stable for at least 8 weeks.     

Efficient synthesis of (±)-8,14-cedranoxide using electrochemical method as a key step

(±)-8,14-Cedranoxide has been synthesized starting from 3,4-dimethoxyphenol, wherein 6-acetoxymethyl-2,6-dimethyl-9-methoxytricyclo[5.3.1.0^1,5]undec-9-en-8,11-dione as a key intermediate has been produced efficiently by means of electrochemical methods.