scholarly journals Separation of Aeruginosin-865 from Cultivated Soil Cyanobacterium (Nostoc sp.) by Centrifugal Partition Chromatography combined with Gel Permeation Chromatography

2015 ◽  
Vol 10 (10) ◽  
pp. 1934578X1501001
Author(s):  
José Cheel ◽  
Mirjana Minceva ◽  
Petra Urajová ◽  
Rabya Aslam ◽  
Pavel Hrouzek ◽  
...  
Keyword(s):  
Gel Permeation Chromatography ◽  
Solvent System ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Lower Phase ◽  
Revolution Speed ◽  
Cultivated Soil ◽  
Gel Permeation ◽  
Isolated Compound ◽  
Soil Cyanobacteria

Aeruginosin-865 was isolated from cultivated soil cyanobacteria using a combination of centrifugal partition chromatography (CPC) and gel permeation chromatography. The solubility of Aer-865 in different solvents was evaluated using the conductor-like screening model for real solvents (COSMO-RS). The CPC separation was performed in descending mode with a biphasic solvent system composed of water-n-BuOH-acetic acid (5:4:1, v/v/v). The upper phase was used as a stationary phase, whereas the lower phase was employed as a mobile phase at a flow rate of 10 mL/min. The revolution speed and temperature of the separation column were 1700 rpm and 25°C, respectively. Preparative CPC separation followed by gel permeation chromatography was performed on 50 mg of crude extract yielding Aer-865 (3.5 mg), with a purity over 95% as determined by HPLC. The chemical identity of the isolated compound was confirmed by comparing its spectroscopic data (UV, HRESI-MS, HRESI-MS/MS) with those of an authentic standard and data available in the literature.

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 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.

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.

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

Article

1998 ◽  
Vol 76 (11) ◽  
pp. 1699-1706 ◽  
Author(s):  
Jonathan Z Knaul ◽  
Mohammad R Kasaai ◽  
V Tam Bui ◽  
Katherine AM Creber
Keyword(s):  
Molecular Weight ◽  
Average Molecular Weight ◽  
Gel Permeation Chromatography ◽  
Solvent System ◽  
Similar Degree ◽  
Degree Of Deacetylation ◽  
Molecular Weights ◽  
Good Accord ◽  
Gel Permeation ◽  
Weight Analysis

Starting from a chitosan sample with a degree of deacetylation of 71%, three separate sample sets were generated by successive deacetylation and reacetylation processes. The degree of deacetylation of samples was determined by UV spectrometry supported by thermogravimetric analysis. The molecular weight of chitosan samples was determined in a solvent system of 0.25 M CH3COOH/0.25 M CH3COONa, using viscometry and gel permeation chromatography (GPC) with a TSK-gel column. The first set of samples had a similar degree of deacetylation (DDA) but differing molecular weights. The second set of samples had a similar molecular weight but differing degrees of deacetylation. The Mark-Houwink-Sakurada constants used for the determination of viscosity average molecular weight and the universal calibration of the GPC system were K = 1.40 × 10-4 dL/g and a = 0.83. Results showed that molecular weights determined from both techniques are in good accord only at lower degrees of deacetylation. This may be attributed to the fact that the chemical structure of chitosan samples could have been largely altered with increasing or decreasing degree of deacetylation. Nevertheless, the trend with which the molecular weights vary with the deacetylation time is consistent over a limited DDA range. A literature review of molecular weight analysis of chitosan is included.Key words: chitosan, degree of deacetylation, gel permeation chromatography, molecular weight, viscometry.

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.

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