RP-HPLC法分离异黄酮中移动相条件的优化

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RP-HPLC法分离异黄酮中移动相条件的优化

郑金珠; 盧敬昊

Center for Advanced Bioseparation Technology and Department of Chemical Engineering, Inha University, 253 Yonghyun-Dong, Nam-Ku, Incheon 402-751, South Korea

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-04-28 发布日期:2007-04-28
通讯作者: 郑金珠

Optimum mobile phase condition for resolving isoflavones in RP-HPLC

ZHENGJinzhu;ROWKyungho

Center for Advanced Bioseparation Technology and Department of Chemical Engineering, Inha University, 253 Yonghyun-Dong, Nam-Ku, Incheon 402-751, South Korea

Received:1900-01-01 Revised:1900-01-01 Online:2007-04-28 Published:2007-04-28
Contact: ZHENG Jinzhu

摘要/Abstract

摘要： In reversed-phase high performance liquid chromatography (RP-HPLC), the mobile phase condition for separating eight isoflavones (daidzin, glycitin, genistin, 6′-o-acetyl daidzin, 6′-o-malonyl genistin, daidzein, glycitein and genistein) was optimized using the HCI (High-Purity Separation Laboratory, Department of Chemical Engineering, Inha University) program software. The optimum composition of mobile phase for the separation of the eight isoflavones was obtained. The elution profiles were calculated by the plate theory based on the equations of retention factor, ln k＝A+BF+CF2, where F was the volume percentage of acetonitrile with 0.1% acetic acid (AA). The first mobile phase composition was water with 0.1% AA/acetonitrile with 0.1% AA (88%/12%, by volume), followed at 9min later by the second composition of mobile phase which was step-changed to 85%/15%, at 19min by the third composition which was step-changed to 73%/27%, at 30min when it was changed to 65%/35% and finally it was maintained in isocratic mode to the end of the run time at 50min. Although, using step gradient mode to separate the isoflavones, the calculated and experimented data were not achieved very good agreement, we could estimate the closed retention time before experiment. And the agreement between the experimental data and the calculated values was relatively good using isocratic separation for eight isoflavones, but the retention time is very long.

关键词: HCI program;optimum;separation condition;isoflavone;HPLC

Abstract: In reversed-phase high performance liquid chromatography (RP-HPLC), the mobile phase condition for separating eight isoflavones (daidzin, glycitin, genistin, 6′-o-acetyl daidzin, 6′-o-malonyl genistin, daidzein, glycitein and genistein) was optimized using the HCI (High-Purity Separation Laboratory, Department of Chemical Engineering, Inha University) program software. The optimum composition of mobile phase for the separation of the eight isoflavones was obtained. The elution profiles were calculated by the plate theory based on the equations of retention factor, ln k＝A+BF+CF2, where F was the volume percentage of acetonitrile with 0.1% acetic acid (AA). The first mobile phase composition was water with 0.1% AA/acetonitrile with 0.1% AA (88%/12%, by volume), followed at 9min later by the second composition of mobile phase which was step-changed to 85%/15%, at 19min by the third composition which was step-changed to 73%/27%, at 30min when it was changed to 65%/35% and finally it was maintained in isocratic mode to the end of the run time at 50min. Although, using step gradient mode to separate the isoflavones, the calculated and experimented data were not achieved very good agreement, we could estimate the closed retention time before experiment. And the agreement between the experimental data and the calculated values was relatively good using isocratic separation for eight isoflavones, but the retention time is very long.

Key words: HCI program, optimum, separation condition, isoflavone, HPLC

郑金珠, ᪋,敬昊. RP-HPLC法分离异黄酮中移动相条件的优化[J]. CIESC Journal.

ZHENGJinzhu,ROWKyungho. Optimum mobile phase condition for resolving isoflavones in RP-HPLC[J]. .

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RP-HPLC法分离异黄酮中移动相条件的优化

Optimum mobile phase condition for resolving isoflavones in RP-HPLC

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