A novel ellipsoid column was designed for centrifugal counter-current chromatography. Performance of the ellipsoid column with a capacity of 3.4 mL was examined with three different solvent systems composed of 1-butanol-acetic acid-water (4:1:5, v/v) (BAW), hexane-ethyl acetate-methanol-0.1 M HCl (1:1:1:1, v/v) (HEMH), and 12.5% (w/w) PEG1000 and 12.5% (w/w) dibasic potassium phosphate in water (PEG-DPP) each with suitable test samples. In dipeptide separation with BAW system, both stationary phase retention (Sf) and peak resolution (Rs) of the ellipsoid column were much higher at 0 column angle (column axis parallel to the centrifugal force) than at 90 column angle (column axis perpendicular to the centrifugal force), where elution with the lower phase at a low flow rate produced the best separation yielding Rs at 2.02 with 27.8% Sf at a flow rate of 0.07 ml/min. In the DNP-amino acid separation with HEMW system, the best results were obtained at a flow rate of 0.05 ml/min with 31.6% Sf yielding high Rs values at 2.16 between DNP-DL-glu and DNP-β-ala peaks and 1.81 between DNP-β-ala and DNP-L-ala peaks. In protein separation with PEG-DPP system, lysozyme and myolobin were resolved at Rs of 1.08 at a flow rate of 0.03 ml/min with 38.9% Sf. Most of those Rs values exceed those obtained from the figure-8 column under similar experimental conditions previously reported. Keywords: ellipsoid column, hydrostatic counter-current chromatographic system, centrifugal counter-current chromatography, retention of the stationary phase, peak resolution, dipeptides, DNP-amino acids, proteins

Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2012
Total Cost
$72,813
Indirect Cost
Name
National Heart, Lung, and Blood Institute
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Shibusawa, Yoichi; Yanagida, Akio; Ogihara, Atsushi et al. (2012) Separation of nucleobases and their derivatives with organic-high ionic strength aqueous phase systems by spiral high-speed counter-current chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 891-892:94-7
Kitazume, Eiichi; Koikawa, Saki; Hui, Lu et al. (2012) Sequential determination of anionic-type detergents by complexation with methylene blue using dual high speed counter-current chromatography. J Chromatogr A 1236:148-51
Zhao, Wenna; He, Jiao; Zhang, Yongmin et al. (2012) Preparative isolation and purification of alkaloids from Picrasma quassiodes (D. Don) Benn. by high-speed countercurrent chromatography. J Liq Chromatogr Relat Technol 35:1597-1606
Cao, Xueli; Yang, Chunlei; Pei, Hairun et al. (2012) Application of counter-current chromatography as a new pretreatment method for the determination of polycyclic aromatic hydrocarbons in environmental water. J Sep Sci 35:596-601
Weisz, Adrian; Witten, Jacob J; Zeng, Yun et al. (2012) Preparation of two novel monobrominated 2-(2',4'-dihydroxybenzoyl)-3,4,5,6-tetrachlorobenzoic acids and their separation from crude synthetic mixtures using vortex counter-current chromatography. J Chromatogr A 1237:106-14
Li, Kang; Zhang, Hongwu; Xie, Huichun et al. (2011) PREPARATIVE ISOLATION AND PURIFICATION OF FIVE FLAVONOIDS FROM POGOSTEMON CABLIN BENTH BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY AND PREPARATIVE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY. J Liq Chromatogr Relat Technol 34:1617-1629
Wang, Xiaohong; Liang, Yong; Peng, Cuilin et al. (2011) PREPARATIVE ISOLATION AND PURIFICATION OF CHEMICAL CONSTITUENTS OF BELAMCANDA BY MPLC, HSCCC AND PREP-HPLC. J Liq Chromatogr Relat Technol 34:241-257
Cao, Xueli; Wang, Qiaoe; Li, Yan et al. (2011) Isolation and purification of series bioactive components from Hypericum perforatum L. by counter-current chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 879:480-8
Weisz, Adrian; Mazzola, Eugene P; Ito, Yoichiro (2011) Preparative separation of 1,3,6-pyrenetrisulfonic acid trisodium salt from the color additive D&C Green No. 8 (pyranine) by pH-zone-refining counter-current chromatography. J Chromatogr A 1218:8249-54
Shinomiya, Kazufusa; Kobayashi, Hiroko; Inokuchi, Norio et al. (2011) Partition Efficiency of High-Pitch Locular Multilayer Coil for Countercurrent Chromatographic Separation of Proteins Using Small-Scale Cross-Axis Coil Planet Centrifuge and Application to Purification of Various Collagenases with Aqueous-Aqueous Polymer P J Liq Chromatogr Relat Technol 34:182-194

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