The performance of type-I high-speed counter-current chromatography was evaluated by changing the column inclination against the rotating centrifugal force field and the core shape of the coiled column. The separations were performed with two different solvent systems composed of 1-butanol-acetic acid-water (4.75:0.25:5, v/v) (BAW) and hexane-ethyl acetate-methanol-0.1 M HCl (1:1:1:1, v/v) (HEMW) using dipeptides and DNP-amino acid as test samples, respectively. In the first series of experiments, a set of short coiled columns connected in series is mounted around the holder hub in two different ways: in the parallel orientation, all column units are arranged in parallel to each other and mounted on the holder at various angles against the horizontal plane. In the zigzag configuration, the neighboring units of the same column are mounted symmetrically forming various angles apart. In the parallel configuration, for both the BAW and HEMW systems, the retention of stationary phase first increased as the column angle decreased from 90 to 60 and then decreased, as the column angle further decreased from 60 to 0, while Rs (peak resolution) continually declined over the entire column angle range from 90 to 0. But, for both solvent systems, with the zigzag configuration, retention of stationary phase and resolution both decreased as the column angle decreased from 90 to 0. In general, Sf and Rs for separation of dipeptides in the BAW system, from 90 to 15, is better for the parallel orientation than for the zigzag configuration. However, at 0, Sf and Rs are better for the zigzag orientation. In the DNP-amino acid separation with the HEMW system, retention of the stationary phase and Rs for the parallel orientation is better than that for the zigzag orientation from 90 to 30, whereas from 30 to 0 the results are opposite. Over all results of our studies revealed that the formally used column orientation at 90 degree inclination yields the highest peak resolution in both solvent systems. In the second series of experiments,three types of novel coiled column configurations, i.e., a triangular coiled column and two elliptical coiled columns were mounted on the holder of type-I countercurrent chromatograph and their performances were evaluated with the above two solvent systems and test samples. The overall results indicated that the performance of compact type-I counter-current chromatography was improved by elliptical coiled column which was mounted with its maximum coil diameter perpendicular to the surface of the column holder.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2011
Total Cost
$74,075
Indirect Cost
Name
National Heart, Lung, and Blood Institute
Department
Type
DUNS #
City
State
Country
Zip Code
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

Showing the most recent 10 out of 32 publications