Continuous Industrial-Scale Centrifugal Partition Chromatography with Automatic Solvent System Handling: Concept and Instrumentation

2020 ◽  
Vol 24 (11) ◽  
pp. 2676-2688
Author(s):  
László Lorántfy ◽  
Dóra Rutterschmid ◽  
Róbert Örkényi ◽  
Dávid Bakonyi ◽  
József Faragó ◽  
...  
Keyword(s):  
Solvent System ◽  
Industrial Scale ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography

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 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 Isolation of sesquiterpenoids from Matricaria chamomilla by means of solvent assisted flavor evaporation and centrifugal partition chromatography

2021 ◽  
Author(s):  
Benedikt Slavik ◽  
Simon Roehrer ◽  
Helene M. Loos ◽  
Mirjana Minceva ◽  
Andrea Buettner
Keyword(s):  
Liquid Chromatography ◽  
Solvent System ◽  
Size Exclusion ◽  
Gas Chromatography Mass Spectrometry ◽  
Volatile Fraction ◽  
Resonance Spectroscopy ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Matricaria Chamomilla ◽  
Exclusion Chromatography

AbstractThe (semi)volatile fraction of Matricaria chamomilla L., an annual herbal plant from the family of Asteraceae, contains high quantities of sesquiterpenes and sesquiterpenoids. A method was developed to achieve isolation and separation of these compounds, using a combination of solvent assisted flavor evaporation (SAFE) and solid support-free liquid-liquid chromatography. The biphasic liquid solvent system n-heptane/ethyl acetate/methanol/water, 5/2/5/2 v/v/v/v (Arizona S) was elaborated as a suitable solvent system for the simultaneous separation of the target compounds. The lab-scale liquid-liquid chromatography separation performed in a countercurrent chromatography (CCC) column was successfully transferred to a semi-preparative centrifugal partition chromatography (CPC) column, which enabled the isolation of artemisia ketone, artemisia alcohol, α-bisabolone oxide A, and (E)-en-yn-dicycloether. α-Bisabolol oxide A and (Z)-en-yn-dicycloether co-eluted, but were successfully separated by subsequent size-exclusion chromatography (SEC). Similarly, spathulenol and α-bisabolol oxide B were obtained as a mixture, and were separated by means of column chromatography using silica gel as stationary phase. The isolated compounds were characterized by means of nuclear magnetic resonance spectroscopy (NMR) and gas chromatography–mass spectrometry (GC-MS). Graphical abstract

scholarly journals Bioassay-Guided Fractionation of Siparuna glycycarpa n-Butanol Extract with Inhibitory Activity against Influenza A(H1N1)pdm09 Virus by Centrifugal Partition Chromatography (CPC)

Molecules ◽  
2022 ◽  
Vol 27 (2) ◽  
pp. 399
Author(s):  
Carla Monteiro Leal ◽  
Suzana Guimarães Leitão ◽  
Leonardo Luiz Oliveira de Mello ◽  
Isabel de Castro Rangel ◽  
Carlos Vinicius Azevedo da Silva ◽  
...  
Keyword(s):  
Influenza A ◽  
Folk Medicine ◽  
Solvent System ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Lower Phase ◽  
Butanol Extract ◽  
Pdm09 Virus ◽  
Influenza A H1n1 ◽  
Bioassay Guided Fractionation

Siparuna glycycarpa occurs in the Amazon region, and some species of this genus are used in Brazilian folk medicine. A recent study showed the inhibitory effect of this species against influenza A(H1N1)pdm09 virus, and in order to acquire active fractions, a polar solvent system n-butanol-methanol-water (9:1:10, v/v) was selected and used for bioassay-guided fractionation of n-butanol extract by centrifugal partition chromatography (CPC). The upper phase was used as stationary phase and the lower phase as mobile (descending mode). Among the collected fractions, the ones coded SGA, SGC, SGD, and SGO showed the highest antiviral inhibition levels (above 74%) at 100 µg·mL−1 after 24 h of infection. The bioactive fractions chemical profiles were investigated by LC-HRMS/MS data in positive and negative ionization modes exploring the Global Natural Products Social Molecular Networking (GNPS) platform to build a molecular network. Benzylisoquinoline alkaloids were annotated in the fractions coded SGA, SGC, and SGD collected during elution step. Aporphine alkaloids, O-glycosylated flavonoids, and dihydrochalcones in SGO were acquired with the change of mobile phase from lower aqueous to upper organic. Benzylisoquinolinic and aporphine alkaloids as well as glycosylated flavonoids were annotated in the most bioactive fractions suggesting this group of compounds as responsible for antiviral activity.

Two novel solvent system compositions for protected synthetic peptide purification by centrifugal partition chromatography

2014 ◽  
Vol 1337 ◽  
pp. 155-161 ◽  
Author(s):  
Nassima Amarouche ◽  
Matthieu Giraud ◽  
Luciano Forni ◽  
Alessandro Butte ◽  
F. Edwards ◽  
...  
Keyword(s):  
Synthetic Peptide ◽  
Solvent System ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Peptide Purification

scholarly journals Isolation of Three Lycorine Type Alkaloids from Rhodolirium speciosum (Herb.) Ravenna Using pH-Zone-Refinement Centrifugal Partition Chromatography and Their Acetylcholinesterase Inhibitory Activities

Metabolites ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 309
Author(s):  
Diana Isabel Correa ◽  
Edgar Pastene-Navarrete ◽  
Luis Bustamante ◽  
Marcelo Baeza ◽  
Julio Alarcón-Enos
Keyword(s):  
Structure Elucidation ◽  
Solvent System ◽  
Methyl Tert Butyl Ether ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Preparative Separation ◽  
Spectroscopy Analysis ◽  
Butyl Ether ◽  
One Step ◽  
Inhibitory Activities

Preparative separation of three lycorine type alkaloids from Rhodolirum speciosum (Amaryllidaceae) was successfully carried out using pH-zone-refinement centrifugal partition chromatography (CPC) using the solvent system methyl-tert-butyl ether/acetonitrile/water (4:1:5, v/v/v) in descending mode. Using this system, Alkaloid 1 (165.7 mg, 88.2%, purity), 2 (60.1 mg, 97.7% purity) and 3 (12.3 mg, 84.4% purity) were obtained in one step. For structure elucidation, the pure alkaloids were subjected to spectroscopy analysis using nuclear magnetic resonance experiments (1H-NMR, 13C-NMR) and gas chromatography coupled with mass spectrometry (GC-MS). Alkaloids 1, 2, and 3 were identified as 1-O-acetyl-5,6-dehydrolycorine, 1-O-acetyl-lycorine, and 1,2-O-diacetyl-5,6-dehydrolycorine, respectively. The acetylcholinesterase inhibitory activity of these alkaloids was IC50 151.1 μg/mL, IC50 203.5 μg/mL, IC50 470.0 μg/mL, and IC50 17.1 μg/mL, respectively.

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