Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography
A comprehensive linear gradient solvent system for centrifugal partition chromatography (CPC) was developed for the bioassay-guided isolation of natural compounds. The gradient solvent system consisted of three different ternary biphasic solvents types: <i>n</i>-hexane–acetonitrile–water...
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MDPI AG
2020-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/13/3077 |
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| author | Ji Hoon Kim Eun Ju Jung Yun Jung Lee En Mei Gao Ahmed Shah Syed Chul Young Kim |
| author_facet | Ji Hoon Kim Eun Ju Jung Yun Jung Lee En Mei Gao Ahmed Shah Syed Chul Young Kim |
| author_sort | Ji Hoon Kim |
| collection | DOAJ |
| description | A comprehensive linear gradient solvent system for centrifugal partition chromatography (CPC) was developed for the bioassay-guided isolation of natural compounds. The gradient solvent system consisted of three different ternary biphasic solvents types: <i>n</i>-hexane–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>), ethyl acetate–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>), and water-saturated <i>n</i>-butanol–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>). The lower phase of the <i>n</i>-hexane–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>) was used as the stationary phase, while its upper phase, as well as ethyl acetate–acetonitrile–water (10:2:8), and water-saturated <i>n</i>-butanol–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>) were pumped to generate a linear gradient elution, increasing the mobile phase polarity. We used the gradient CPC to identify antioxidant response elements (AREs), inducing compounds from <i>Centipeda minima</i>, using an ARE-luciferase assay in HepG2 cells, which led to the purification of the active molecules 3-methoxyquercetin and brevilin A. The developed CPC solvent systems allow the separation and isolation of compounds with a wide polarity range, allowing active molecule identification in the complex crude extract of natural products. |
| first_indexed | 2024-03-10T18:39:03Z |
| format | Article |
| id | doaj.art-f29d59dc66b74ddf8fd13ae325da2d21 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T18:39:03Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-f29d59dc66b74ddf8fd13ae325da2d212023-11-20T06:00:23ZengMDPI AGMolecules1420-30492020-07-012513307710.3390/molecules25133077Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition ChromatographyJi Hoon Kim0Eun Ju Jung1Yun Jung Lee2En Mei Gao3Ahmed Shah Syed4Chul Young Kim5College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Gyeonggi-do 15588, KoreaCollege of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Gyeonggi-do 15588, KoreaCollege of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Gyeonggi-do 15588, KoreaCollege of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Gyeonggi-do 15588, KoreaDepartment of Pharmacognosy, Faculty of Pharmacy, University of Sindh, Jamshoro 76088, PakistanCollege of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Gyeonggi-do 15588, KoreaA comprehensive linear gradient solvent system for centrifugal partition chromatography (CPC) was developed for the bioassay-guided isolation of natural compounds. The gradient solvent system consisted of three different ternary biphasic solvents types: <i>n</i>-hexane–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>), ethyl acetate–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>), and water-saturated <i>n</i>-butanol–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>). The lower phase of the <i>n</i>-hexane–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>) was used as the stationary phase, while its upper phase, as well as ethyl acetate–acetonitrile–water (10:2:8), and water-saturated <i>n</i>-butanol–acetonitrile–water (10:2:8, <i>v</i>/<i>v</i>) were pumped to generate a linear gradient elution, increasing the mobile phase polarity. We used the gradient CPC to identify antioxidant response elements (AREs), inducing compounds from <i>Centipeda minima</i>, using an ARE-luciferase assay in HepG2 cells, which led to the purification of the active molecules 3-methoxyquercetin and brevilin A. The developed CPC solvent systems allow the separation and isolation of compounds with a wide polarity range, allowing active molecule identification in the complex crude extract of natural products.https://www.mdpi.com/1420-3049/25/13/3077centrifugal partition chromatographycomprehensive linear gradient elutionbioactivity-guided isolation3-methoxyquercetinbrevilin A<i>Centipeda minima</i> |
| spellingShingle | Ji Hoon Kim Eun Ju Jung Yun Jung Lee En Mei Gao Ahmed Shah Syed Chul Young Kim Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography Molecules centrifugal partition chromatography comprehensive linear gradient elution bioactivity-guided isolation 3-methoxyquercetin brevilin A <i>Centipeda minima</i> |
| title | Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography |
| title_full | Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography |
| title_fullStr | Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography |
| title_full_unstemmed | Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography |
| title_short | Bioassay-Guided Separation of <i>Centipeda minima</i> Using Comprehensive Linear Gradient Centrifugal Partition Chromatography |
| title_sort | bioassay guided separation of i centipeda minima i using comprehensive linear gradient centrifugal partition chromatography |
| topic | centrifugal partition chromatography comprehensive linear gradient elution bioactivity-guided isolation 3-methoxyquercetin brevilin A <i>Centipeda minima</i> |
| url | https://www.mdpi.com/1420-3049/25/13/3077 |
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