Mitsunori Washitake

High-performance anion-exchange chromatography of ultraviolet-absorbing constituents of human urine.

A 100-microliters urine sample was chromatographed on a column packed with a strongly basic macroreticular anion-exchange resin (Diaion CDR-10,5--7 micrometers with a nominal 35% cross linkage). The elution was performed with a linear acetate gradient from 0 to 6.0 M at an average flow-rate of 0.72 ml/min and at an average pressure of 104 kg/cm2. The relative standard deviation of retention times and peak height was +/- 4% or less. The properties of the macroreticular anion-exchange resin, the effect of the particle size, the pH of acetate buffers, and the effect of the flow-rate of the eluent on the separation were investigated. Thirty three components of urine were then resolved and named.

High-performance anion-exchange chromatography of human urine using perchlorate gradient elution systems

A 100-microliter volume of urine was chromatographed on a 50 X 0.4 cm I.D. column packed with a macroreticular anion-exchange resin. Elution was performed with a concave ammonium perchlorate gradient from 0 to 0.25 M at a flow-rate of 0.75 ml/min and a pressure of 7.5-11 MPa. With this perchlorate gradient, no baseline drift occurred in the detection at 254 nm, and even detection at 200 nm was possible. The effect of the addition of ethanol or acetonitrile to the ammonium perchlorate solution was investigated. For the assignment of peaks, ultraviolet spectra of the peaks were measured with stopped-flow scanning spectrophotometry.

High-Resolution Anion-Exchange Chromatography of Ultraviolet-Absorbing Constituents of Human Erythrocytes

Abstract The analysis of nucleic acid components of human erythrocytes was achieved by high-resolution anion exchange chromatography using a column packed with a macroreticular anion-exchange resin and linear gradient elution with ammonium acetate solution, pH 4.5. On the chromatogram of trichloroacetic acid extracts from human erythrocytes, at least 50 components were detected as ultraviolet-absorbing constituents. On assignment of the chromatographic peaks they were found to be nucleic acid components. The analysis was achieved in 120 min and the elution time of guanosine 5′-triphosphate was 64.3 min.