Title: | Counter-Current Chromatographic Separation of Nucleic Acid Constituents with a Polar Volatile Organic-Aqueous Two-Phase Solvent Systems with ELSD Detection |
Author(s): | Shibusawa Y; Morikawa G; Yanagida A; Ito Y; |
Address: | "Division of Pharmaceutical and Biomedical Analysis, School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan. Bioseparation Technology, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA" |
DOI: | 10.2174/1874065001206010009 |
ISSN/ISBN: | 1874-0650 (Print) 1874-0650 (Electronic) 1874-0650 (Linking) |
Abstract: | "Highly hydrophilic volatile organic/aqueous two-phase solvent systems containing an organic salt such as, acetonitrile/800 mM and 1200 mM ammonium acetate (1 : 1, v/v) were efficiently utilized for high-speed counter-current chromatography (HSCCC) to separate hydrophilic compounds. The retention of the upper and the lower stationary phases in the column of the cross-axis coil planet centrifuge (CCC instrument) was studied by changing the flow-rate of the mobile phase (1.0-3.0 ml/min). Using the acetonitrile/800 mM ammonium acetate two-phase solvent system, the stationary phase was retained at 46.3% relative to the total column capacity of 65 ml by the reversed-phase elution mode at a flow-rate of 1.0 ml/min. The best retention of the stationary upper phase of 51.5% was obtained by the solvent system of the acetonitrile/1200 mM ammonium acetate at the above flow-rate. With the acetonitrile/800 mM ammonium acetate system the base line separation of adenine and adenosine monophosphate (AMP) detected by evaporative light scattering detector (ELSD) and UV was achieved with lower phase mobile at a flow-rate of 1.0 ml/min within 70 min" |
Keywords: | Amp Elsd Hsccc hydrophilic compounds; |
Notes: | "PubMed-not-MEDLINEShibusawa, Yoichi Morikawa, Go Yanagida, Akio Ito, Yoichiro eng Z99 HL999999/Intramural NIH HHS/ Netherlands 2012/01/01 Open Anal Chem J. 2012; 6:9-14. doi: 10.2174/1874065001206010009" |