Chromatographic separation of galactosylkojic acid
The mechanism of the separation of galactosylkojic acid (GKA) produced by the reaction of immobilized β-galactosidase with kojic acid (KA) and lactose as substrates using Toyopearl HW40 size exclusion chromatography was investigated. The effects of the mobile phase conditions (salt concentration and...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier Sci B.V.Amsterdam
1997
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/114093/1/114093.pdf |
| _version_ | 1824452362828251136 |
|---|---|
| author | Yamamoto, Shuichi Nakanishi, Kazuhiro Hassan, Mohd Ali |
| author_facet | Yamamoto, Shuichi Nakanishi, Kazuhiro Hassan, Mohd Ali |
| author_sort | Yamamoto, Shuichi |
| collection | UPM |
| description | The mechanism of the separation of galactosylkojic acid (GKA) produced by the reaction of immobilized β-galactosidase with kojic acid (KA) and lactose as substrates using Toyopearl HW40 size exclusion chromatography was investigated. The effects of the mobile phase conditions (salt concentration and pH) on the distribution coefficient K for GKA and KA determined from the peak retention volume were investigated. The K values for GKA and KA decreased slightly when the pH in the mobile phase was increased from 6 to 7, while a further increase in the pH from 7 to 8 caused a considerable decrease in the K values. The K values increased with increasing NaCl concentration (0 to 0.5 M), especially at pH 8. The K value for GKA at pH 8 was almost the same as that for neutral sugar having the same molecular weight. As the gels have negative charges at pH 6-8 and the dissociation constant of KA is ca. 10-8, these results indicate that the distribution of GKA and KA is governed by hydrophobic interaction adsorption and electrostatic repulsion. Although complete baseline separation of GKA was achieved under any of the conditions examined, on the basis of the plate height and K value data the optimum mobile phase conditions were determined to be pH 7 and 50 mM NaCl.; The mechanism of the separation of galactosylkojic acid (GKA) produced by the reaction of immobilized β-galactosidase with kojic acid (KA) and lactose as substrates using Toyopearl HW40 size exclusion chromatography was investigated. The effects of the mobile phase conditions (salt concentration and pH) on the distribution coefficient K for GKA and KA determined from the peak retention volume were investigated. The K values for GKA and KA decreased slightly when the pH in the mobile phase was increased from 6 to 7, while a further increase in the pH from 7 to 8 caused a considerable decrease in the K values. The K values increased with increasing NaCl concentration (0 to 0.5 M), especially at pH 8. The K value for GKA at pH 8 was almost the same as that for neutral sugar having the same molecular weight. As the gels have negative charges at pH 6-8 and the dissociation constant of KA is ca. 10-8, these results indicate that the distribution of GKA and KA is governed by hydrophobic interaction adsorption and electrostatic repulsion. Although complete baseline separation of GKA was achieved under any of the conditions examined, on the basis of the plate height and K value data the optimum mobile phase conditions were determined to be pH 7 and 50 mM NaCl. |
| first_indexed | 2025-02-19T02:49:19Z |
| format | Article |
| id | upm.eprints-114093 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2025-02-19T02:49:19Z |
| publishDate | 1997 |
| publisher | Elsevier Sci B.V.Amsterdam |
| record_format | dspace |
| spelling | upm.eprints-1140932024-12-10T01:43:35Z http://psasir.upm.edu.my/id/eprint/114093/ Chromatographic separation of galactosylkojic acid Yamamoto, Shuichi Nakanishi, Kazuhiro Hassan, Mohd Ali The mechanism of the separation of galactosylkojic acid (GKA) produced by the reaction of immobilized β-galactosidase with kojic acid (KA) and lactose as substrates using Toyopearl HW40 size exclusion chromatography was investigated. The effects of the mobile phase conditions (salt concentration and pH) on the distribution coefficient K for GKA and KA determined from the peak retention volume were investigated. The K values for GKA and KA decreased slightly when the pH in the mobile phase was increased from 6 to 7, while a further increase in the pH from 7 to 8 caused a considerable decrease in the K values. The K values increased with increasing NaCl concentration (0 to 0.5 M), especially at pH 8. The K value for GKA at pH 8 was almost the same as that for neutral sugar having the same molecular weight. As the gels have negative charges at pH 6-8 and the dissociation constant of KA is ca. 10-8, these results indicate that the distribution of GKA and KA is governed by hydrophobic interaction adsorption and electrostatic repulsion. Although complete baseline separation of GKA was achieved under any of the conditions examined, on the basis of the plate height and K value data the optimum mobile phase conditions were determined to be pH 7 and 50 mM NaCl.; The mechanism of the separation of galactosylkojic acid (GKA) produced by the reaction of immobilized β-galactosidase with kojic acid (KA) and lactose as substrates using Toyopearl HW40 size exclusion chromatography was investigated. The effects of the mobile phase conditions (salt concentration and pH) on the distribution coefficient K for GKA and KA determined from the peak retention volume were investigated. The K values for GKA and KA decreased slightly when the pH in the mobile phase was increased from 6 to 7, while a further increase in the pH from 7 to 8 caused a considerable decrease in the K values. The K values increased with increasing NaCl concentration (0 to 0.5 M), especially at pH 8. The K value for GKA at pH 8 was almost the same as that for neutral sugar having the same molecular weight. As the gels have negative charges at pH 6-8 and the dissociation constant of KA is ca. 10-8, these results indicate that the distribution of GKA and KA is governed by hydrophobic interaction adsorption and electrostatic repulsion. Although complete baseline separation of GKA was achieved under any of the conditions examined, on the basis of the plate height and K value data the optimum mobile phase conditions were determined to be pH 7 and 50 mM NaCl. Elsevier Sci B.V.Amsterdam 1997 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/114093/1/114093.pdf Yamamoto, Shuichi and Nakanishi, Kazuhiro and Hassan, Mohd Ali (1997) Chromatographic separation of galactosylkojic acid. Journal of Fermentation and Bioengineering, 84 (1). pp. 82-85. ISSN 0922-338X; eISSN: 0922-338X https://linkinghub.elsevier.com/retrieve/pii/S0922338X97827912 10.1016/s0922-338x(97)82791-2 |
| spellingShingle | Yamamoto, Shuichi Nakanishi, Kazuhiro Hassan, Mohd Ali Chromatographic separation of galactosylkojic acid |
| title | Chromatographic separation of galactosylkojic acid |
| title_full | Chromatographic separation of galactosylkojic acid |
| title_fullStr | Chromatographic separation of galactosylkojic acid |
| title_full_unstemmed | Chromatographic separation of galactosylkojic acid |
| title_short | Chromatographic separation of galactosylkojic acid |
| title_sort | chromatographic separation of galactosylkojic acid |
| url | http://psasir.upm.edu.my/id/eprint/114093/1/114093.pdf |
| work_keys_str_mv | AT yamamotoshuichi chromatographicseparationofgalactosylkojicacid AT nakanishikazuhiro chromatographicseparationofgalactosylkojicacid AT hassanmohdali chromatographicseparationofgalactosylkojicacid |