Three-phase solvent systems for comprehensive separation of a wide variety of compounds by high-speed counter-current chromatography.
Three-phase solvent systems were efficiently utilized for high-speed counter-current chromatography (HSCCC) to separate multiple components with a wide range of hydrophobicity. The compositions of three-phase systems were optimized according to their physical parameters such as volume ratio, viscosity and specific gravity of upper (UP), middle (MP) and lower (LP) phases. The three-phase systems composed of n-hexane-methyl acetate-acetonitrile-water (4:4:3:4, v/v/v/v) was selected for HSCCC separation of a mixture of 15 standard compounds with a wide range in hydrophobicity from beta-carotene to tryptophan. The separation was initiated by filling the column with a mixture of MP and LP both as a stationary phase followed by elution with UP to separate the hydrophobic compounds. Then the mobile phase was switched to MP to elute the moderately hydrophobic compounds, and finally the polar compounds still retained in the column were fractionated by eluting the column with LP. The system successfully resolved all 15 compounds in one-step operation in 70 min.
Shibusawa
Yoichi
Y
Tokyo University of Pharmacy and Life Science, Division of Structural Biology and Analytical Science, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan. sibusawa@ps.toyaku.ac.jp
Yamakawa
Yutaka
Y
Noji
Ryoko
R
Yanagida
Akio
A
Shindo
Heisaburo
H
Ito
Yoichiro
Y
D002853:Chromatography, Liquid / Q000379:methods*; D003377:Countercurrent Distribution / Q000379:methods*; D015394:Molecular Structure; D009930:Organic Chemicals / Q000032:analysis* / Q000737:chemistry; D012997:Solvents / Q000737:chemistry*
D009930:Organic Chemicals; D012997:Solvents