Motoko Takeda

Fluorescene high-performance liquid chromatography of reducing using Dns-hydrazie as a pre-labelling reagent

Abstract The fluorescence high-performance liquid chromatography of reducing sugars is describe. Dns-hydrazine was used as a fluorescent pre-labelling reagent. Various reducing sugars, such as fucose, glucose, galactose and maltose, were labelled with Dns-hydrazine in trichloroacetic acid—ethanol solution and then chromatographed on LiChrosorb SI 100 column using ethanol—water—chloroform as eluent. Good separations of thirteen reducing sugars involving pentoses, hexoses and disaccharides were obtained within 25 min by stepwise elutin system. Calibration graphs are linear in the range from 0.05 to 1.0 nmol for each sugar. The detection limits were about 3– 20 pmol of reducing sugar.

A fluorophotometric method for the determination of amino acids by using pyridoxal and zinc(II) ion

Abstract A fluorophotometric method for the determination of α-oxo acids has been developed based on their interaction with pyridoxamine and Zn(II) ion in pyridine-methanol solution to yield highly fluorescent chelates. This fluorophotometric procedure is simple and highly sensitive to α-oxo acids. No detectable interference for the determination of pyruvate occurred with any of the amino acids, amines, sugars, organic acids, and inorganic salts. Recoveries for pyruvate added to bloods and serum ranged from 90.1 to 103.2%. As little as 5 × 10−11 mol of α-oxo acid may be measured.