scholarly journals Bioassay-Guided Isolation of Two Eudesmane Sesquiterpenes from Lindera strychnifolia Using Centrifugal Partition Chromatography

Molecules ◽  
2021 ◽  
Vol 26 (17) ◽  
pp. 5269
Author(s):  
Ji Hoon Kim ◽  
Je-Seung Jeon ◽  
Jung Hoon Kim ◽  
Eun Ju Jung ◽  
Yun Jung Lee ◽  
...  
Keyword(s):  
Solvent System ◽  
Preparative Hplc ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Dual Mode ◽  
Two Phase ◽  
Chemical Structures ◽  
High Recovery Rate ◽  
Responsive Element ◽  
Antioxidant Responsive Element

In this study, a centrifugal partition chromatography (CPC) separation was applied to identify antioxidant-responsive element (ARE) induction molecules from the crude extract of Lindera strychnifolia roots. CPC was operated with a two-phase solvent system composed of n-hexane-methanol-water (10:8.5:1.5, v/v/v) in dual mode (descending to ascending), which provided a high recovery rate (>95.5%) with high resolution. Then, ARE induction activity of obtained CPC fractions was examined in ARE-transfected HepG2 cells according to the weight ratios of the obtained fractions. The fraction exhibiting ARE-inducing activity was further purified by preparative HPLC that led to isolation of two eudesmane type sesquiterpenes as active compounds. The chemical structures were elucidated as linderolide U (1) and a new sesquiterpene named as linderolide V (2) by spectroscopic data. Further bioactivity test demonstrated that compounds 1 and 2 enhanced ARE activity by 22.4-fold and 7.6-fold, respectively, at 100 μM concentration while 5 μM of sulforaphane induced ARE activity 24.8-fold compared to the control.

scholarly journals Preparative and Rapid Purification of Saponins from Asparagus racemosus Root by High Performance Centrifugal Partition Chromatography

2017 ◽  
Vol 12 (2) ◽  
pp. 1934578X1701200 ◽  
Author(s):  
Churanya Onlom ◽  
Yi Yang ◽  
Haji A. Aisa ◽  
Neti Woranuch ◽  
Watoo Phrompittayarat ◽  
...  
Keyword(s):  
High Performance ◽  
Human Fibroblasts ◽  
Gel Filtration ◽  
Solvent System ◽  
Asparagus Racemosus ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Two Phase ◽  
Isolation And Purification ◽  
Biological Studies

High performance centrifugal partition chromatography (HPCPC) was applied to the rapid isolation and purification of saponin glycosides in Asparagus racemosus Willd. root. A two-phase solvent system composed of CHCl3-MeOH-water (4:4:2, v/v) in descending mode was used for the separation, yielding shatavarin IX (1) and asparacoside (2) in one step. Asparanin A (3) and shatavarin V (4) were separated by repeated HPCPC fractionation using CH2Cl2-MeOH-water (4:4:2, v/v) as the solvent system, followed by either gel-filtration or TLC. Their structures were identified by NMR spectroscopy and ESI/MS. The A. racemosus extracts and 1, 2, 3 and 4 were cytotoxic towards human hepato- and prostate-carcinoma cell lines (IC50 14–37 μM), while primary human fibroblasts were less vulnerable (IC50 22–66 μM), i.e., every saponin glycoside showed selectivity towards carcinoma cells compared with normal fibroblasts. HPCPC has proven rapidity to separate complex mixtures of phytochemicals yielding quantities suited to biological studies.

scholarly journals Preparative Separation of Three Monoterpenes from Perilla frutescens var. crispa Using Centrifugal Partition Chromatography

2019 ◽  
Vol 2019 ◽  
pp. 1-7
Author(s):  
Bomi Nam ◽  
Sunil Babu Paudel ◽  
Jin-Baek Kim ◽  
Chang Hyun Jin ◽  
Dongho Lee ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
Mobile Phase ◽  
Solvent System ◽  
Perilla Frutescens ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Drying Process ◽  
Two Phase ◽  
C Nmr

Three monoterpenes, namely, 9-hydroxy isoegomaketone (1), isoegomaketone (2), and perilla ketone (3), were successfully separated from the supercritical carbon dioxide (SC-CO2) extract of the leaves of Perilla frutescens var. crispa (cv. Antisperill; Lamiaceae) by centrifugal partition chromatography (CPC). To obtain large quantities of these materials required for studies on their mechanism of action and in vivo effectiveness in inflammation, we used CPC because of its high loading capacity and reproducibility to purify the three compounds. Compound 1 (2.60 mg, 96.7% purity at 254 nm) was purified from 500 mg of the SC-CO2 extract of P. frutescens var. crispa (cv. Antisperill), using a two-phase solvent system comprising n-hexane/ethyl acetate/ethanol/water (5:5:5:5 v/v) in a descending mode. As compounds 2 (56.1 mg, 97.6% purity at 254 nm) and 3 (78.6 mg, 96.1% purity at 254 nm) are highly volatile and difficult to recover from an aqueous mobile phase after purification during the drying process, they were obtained from the same amount of the processed extract in an ascending mode using the upper organic phase as the mobile phase (n-hexane/ethyl acetate/ethanol/water, 8:2:8:2 v/v). The structures of compounds 1–3 were confirmed by 1H- and 13C-NMR analysis. Thus, based on our findings, we recommend centrifugal partition chromatography as a powerful technique for purifying the active principal compounds 1 and 2 from the leaves of P. frutescens var. crispa.

scholarly journals Application of Linear Gradient Solvent System in Centrifugal Partition Chromatography Facilitating Bioassay-Guided Fractionation of Yongdamsagan-Tang, Traditional Oriental Decoction

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Ji Hoon Kim ◽  
Eun Ju Jung ◽  
Yun Jung Lee ◽  
Chul Young Kim ◽  
Je-Seung Jeon
Keyword(s):  
Natural Products ◽  
Crude Extract ◽  
Solvent System ◽  
Bioactive Molecules ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Isolation And Identification ◽  
Time Saving ◽  
K Value ◽  
Linear Gradient

As important pharmaceutical resources, traditional herbal medicines retain continuous attention. To do that, isolation and identification of bioactive molecules from traditional herbal decoction are important. However, conventional fractionation through octadecyl silica column faces irreversible sample adsorption that causes a bias in bioactivity assessment. However, liquid-liquid chromatographic system suffers tedious K value calculation as well as insufficient capacity in separation power when crude extract composed of widely ranging polarities. Here, we developed a comprehensive linear gradient solvent system for centrifugal partition chromatography (CPC) to aid bioassay-guided isolation. The lower aqueous phase of the n-hexane-acetonitrile-water (10:2:8, v/v) was used as the stationary, whereas its upper organic phase followed by the upper phase of ethyl acetate-acetonitrile-water and water-saturated n-butanol-acetonitrile-water in the same ratio were eluted in a linear gradient mode, thereby increasing polarity in the mobile phase. The HPLC profiling of CPC fraction showed that proposed gradient CPC was suitable to separate metabolites from Yongdamsagan-Tang, a traditional medicinal decoction made of ten herbal plants. Exhibiting a high recovery yield of 98.3%, antioxidant response element (ARE) luciferase-inducing assay in HepG2 cells indicated that the fractions composed of baicalein and wogonin, the marker natural products of Scutellaria baicalensis, were to be the most effective molecules from Yongdamsagan-Tang. The presented results demonstrated that bioassay-guided separation that assisted with a linear gradient CPC is an incomparable alternative to HPLC and biphasic CPC in terms of higher yield rate and redundant K value calculation, respectively, which led to an unbiased/time-saving separation and identification of bioactive molecules from the complex crude extract of natural products.

scholarly journals Bioassay-Guided Separation of Centipeda minima Using Comprehensive Linear Gradient Centrifugal Partition Chromatography

Molecules ◽  
2020 ◽  
Vol 25 (13) ◽  
pp. 3077
Author(s):  
Ji Hoon Kim ◽  
Eun Ju Jung ◽  
Yun Jung Lee ◽  
En Mei Gao ◽  
Ahmed Shah Syed ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
Gradient Elution ◽  
Solvent System ◽  
Luciferase Assay ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Lower Phase ◽  
Linear Gradient ◽  
Centipeda Minima ◽  
Water Saturated

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: n-hexane–acetonitrile–water (10:2:8, v/v), ethyl acetate–acetonitrile–water (10:2:8, v/v), and water-saturated n-butanol–acetonitrile–water (10:2:8, v/v). The lower phase of the n-hexane–acetonitrile–water (10:2:8, v/v) was used as the stationary phase, while its upper phase, as well as ethyl acetate–acetonitrile–water (10:2:8), and water-saturated n-butanol–acetonitrile–water (10:2:8, v/v) 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 Centipeda minima, 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.

scholarly journals Application of Centrifugal Partition Chromatography for Bioactivity-Guided Purification of Antioxidant-Response-Element-Inducing Constituents from Atractylodis Rhizoma Alba

Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2274 ◽  
Author(s):  
Myeong Kim ◽  
Ji Kim ◽  
Ahmed Syed ◽  
Young-Mi Kim ◽  
Kevin Choe ◽  
...  
Keyword(s):  
Antioxidant Response ◽  
Luciferase Reporter ◽  
Antioxidant Response Element ◽  
Partition Chromatography ◽  
Reporter Assay ◽  
Centrifugal Partition Chromatography ◽  
Response Element ◽  
1H And 13C Nmr ◽  
Chemical Structures ◽  
Atractylenolide I

Activity-guided separation of antioxidant response element (ARE)-inducing constituents from the rhizomes of Atractylodis Rhizoma Alba was performed by the combination of centrifugal partition chromatography (CPC) and an ARE luciferase reporter assay. From 3 g of the active n-hexane fraction, one polyacetylene, (6E,12E)-tetradeca-6,12-dien-8,10-diyne-1,3-diyl diacetate (47.3 mg), and two sesquiterpenes, atractylenolide I (40.9 mg), and selina-4(14),7(11)-dien-8-one (6.0 mg) were successfully isolated by CPC with n-hexane–ethyl acetate–methanol–water (8:2:8:2, v/v). The chemical structures of the isolated compounds were determined by 1H- and 13C-NMR and ESI-MS. Among the isolated compounds, (6E,12E)-tetradeca-6,12-diene-8,10-diyne-1,3-diol diacetate and selina-4(14),7(11)-dien-8-one increased ARE activity 32.9-fold and 16.6-fold, respectively, without significant cytotoxicity, when 5 µM sulforaphane enhanced ARE activity 27.1-fold. However, atractylenolide I did not increase ARE activity at 100 µM, and showed cytotoxicity at concentrations over 10 µM.

Sign in / Sign up

Export Citation Format

Share Document