Efficient Counter-Current Chromatographic Isolation and Structural Identification of Two New Cinnamic Acids from Echinacea Purpurea

Two new cinnamic acids, 2- O-caffeoyl-3- O-isoferuloyltartaric (3), and 2, 3-di- O-isoferuloyltartaric acid (5), along with three known caffeic acids, cichoric acid (1), 2- O-caffeoyl-3- O-feruloyltartaric acid (2) and 2- O- caffeoyl-3- O-p-coumaroyltartaric acid (4), have been successfully isolated and purified from Echinacea purpurea. In this study, we investigated an efficient method for the preparative isolation and purification of cinnamic acids from E. purpurea by high-speed counter-current chromatography (HSCCC). The separation was performed using a two-phase solvent composed of n-hexane-ethyl-acetate-methanol-0.5% aqueous acetic acid (1:3:1:4, v/v). The upper phase was used as the stationary phase and the lower phase as the mobile phase, with a flow rate of 1.6 mL/min. From 250 mg of crude extracts, 65.1 mg of 1, 8.3 mg of 2, 4.0 mg of 3, 4.5 mg of 4, and 4.3 mg of 5 were isolated in one-step, with purities of 98.5%, 97.7%, 94.6%, 94.3%, and 98.6%, respectively, as evaluated by HPLC-DAD. The chemical structures were identified by electro spray ionization mass spectrometry (ESI-MS) and one- and two-dimensional NMR spectra. HSCCC was very efficient for the separation and purification of the cinnamic acids from E. purpurea.

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Preparative isolation and purification of seven compounds from Hibiscus mutabilis L. leaves by two-step high-speed counter-current chromatography

Chemical Industry and Chemical Engineering Quarterly ◽

10.2298/ciceq140522034h ◽

2015 ◽

Vol 21 (2) ◽

pp. 331-341 ◽

Cited By ~ 2

Author(s):

Zhuoni Hou ◽

Xianrui Liang ◽

Feng Su ◽

Weike Su

Keyword(s):

Ethyl Acetate ◽

High Speed ◽

Solvent System ◽

Ionization Mass ◽

Two Phase ◽

Purification Method ◽

Isolation And Purification ◽

Chemical Structures ◽

13C Nuclear Magnetic Resonance ◽

Counter Current

Seven compounds from Hibiscus mutabilis L. leaves were first successfully achieved by two-step high-speed counter-current chromatography with two-phase solvent system composed of n-butanol-ethyl acetate-water (1:6:9, v/v/v) and n-hexane-ethyl acetate-methanol-water (3:5:3:5, v/v/v/v/). The critical experimental parameters of first-step separation were optimized with response surface methodology as follows: flow rate was 1.1 mL/min, revolution speed was 800 rpm and temperature was 30?C. Under the optimal conditions, around 5.0 mg of salicylic acid, 13.6 mg of rutin, 5.5 mg of genistein were obtained in 100 mg crude sample. Then, 9.2 mg of potengriffioside A, 4.7 mg of kaempferol 3-O-rutinoside, 3.0 mg of steppogenin and 2.5 mg of emodin were obtained by second-step separation. The purities of the seven compounds determined by UPLC were 96.2%, 93.8%, 95.4%, 94.3%, 98.0%, 94.1% and 90.8%, respectively. Their chemical structures were identified by electron spray ionization mass spectroscopy (ESI-MS) and 1H, 13C nuclear magnetic resonance (NMR). Furthermore, compound steppogenin and genistein were first reported from Hibiscus mutabilis L. The purification method was simple, efficient and evaded tedious separation process.

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Isolation and Identification of Anthocyanin Component in the Fruits of Acanthopanax Sessiliflorus (Rupr. & Maxim.) Seem. by Means of High Speed Counter Current Chromatography and Evaluation of Its Antioxidant Activity

Molecules ◽

10.3390/molecules25081781 ◽

2020 ◽

Vol 25 (8) ◽

pp. 1781 ◽

Cited By ~ 2

Author(s):

Liang Chen ◽

Xiulan Xin ◽

Hui Feng ◽

Shuangshi Li ◽

Qiguang Cao ◽

...

Keyword(s):

Antioxidant Activity ◽

High Speed ◽

Radical Scavenging ◽

Ionization Mass ◽

Anthocyanin Content ◽

Separation And Purification ◽

Isolation And Identification ◽

Antioxidant Power ◽

Acanthopanax Sessiliflorus ◽

Counter Current

Acanthopanax sessiliflorus (Rupr. & Maxim.) Seem. (Araliaceae) is one of the most abundant species of genus Acanthopanax. The fruits of A. sessiliflorus are used in traditional medical protocols as an analgesic, tonic, antidiabetic, antihypertensive, anti-inflammatory, antitumor, and immune-stimulating agent. In this work, we carried out a comprehensive investigation into the anthocyanin components in the fruits of A. sessiliflorus. The anthocyanin content in the fresh fruits of A. sessiliflorus was determined by high performance liquid chromatography-diode array detection (HPLC/DAD), and the anthocyanin component was isolated from these using high-speed counter-current chromatography (HSCCC) and elucidated by electro-spray ionization-mass spectrometry (ESI/MS), 1H- and 13C-NMR. Its antioxidant activity was evaluated by ferric-reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). We found that A. sessiliflorus contained a gross anthocyanin content of 121.35 mg/100 g. HSCCC was successfully used for separation and purification of the primary anthocyanin component, cyanidin 3-xylosyl-galactoside. The antioxidant and radical scavenging tests indicated that cyanidin 3-xylosyl-galactoside is a potent antioxidant.

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Systematic Separation and Purification of Alkaloids from Euchresta tubulosa Dunn. by Various Chromatographic Methods

Processes ◽

10.3390/pr7120924 ◽

2019 ◽

Vol 7 (12) ◽

pp. 924

Author(s):

Wei-Xin Li ◽

Huan Wang ◽

Ai-Wen Dong

Keyword(s):

Natural Products ◽

Silica Gel ◽

Column Chromatography ◽

High Speed ◽

High Performance ◽

Ionization Mass ◽

Separation And Purification ◽

Two Phase ◽

Rapid Separation ◽

Chemical Structures

High-speed countercurrent chromatography (HSCCC) and silica gel column chromatography were used to separate and purify alkaloids from Chinese herbal medicine Euchresta tubulosa Dunn. The purpose of this study is to provide a system mode for rapid separation of alkaloids from natural products. In the experiment, the eluent of silica gel column chromatography was screened by thin layer chromatography (TLC) to obtain four components with different polarity. Then, the two-phase solvent systems of different components were selected and purified by HSCCC. Four alkaloids with relatively high content were obtained by this mode successfully, including matrine (28 mg), oxymatrine (32 mg), N-formyl cytisine (24 mg), and cytisine (58 mg). The purity was higher than 91% by high performance liquid chromatography–ultraviolet (HPLC-UV) and their chemical structures were identified by nuclear magnetic resonance (NMR) and electron ionization mass spectrometry (EI-MS). The results showed that the combination of HSCCC and silica gel column chromatography could make alkaloids from natural products separate systematically.

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Separation of avermectin components from Streptomyces avemitilis extraction using high-speed counter-current chromatography

Chemical Industry and Chemical Engineering Quarterly ◽

10.2298/ciceq120410089s ◽

2013 ◽

Vol 19 (4) ◽

pp. 563-571 ◽

Cited By ~ 1

Author(s):

Weike Su ◽

Zhuoni Hou ◽

Xianrui Liang

Keyword(s):

High Speed ◽

Solvent System ◽

Ionization Mass ◽

Two Phase ◽

Preparative Scale ◽

Chemical Structures ◽

Peak Resolution ◽

Ionization Mass Spectroscopy ◽

13C Nuclear Magnetic Resonance ◽

Counter Current

Three compounds of antibiotics-avermectins from fertilizing product of Streptomyces avemitilis are achieved by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (6:4:5:5, v/v) on a preparative scale. The separation condition was: 1.5 mL/min (0 to 200 min) and 2.0 mL/min (200 to the end), 900 rpm and 20?C based on the peak resolution. About 11.9 mg of avermectin B1a, 1.0 mg of avermectin B1b and 9.6 mg of avermectin B2a from 50 mg of crude extract were obtained by one-step separation. The purities of the three compounds determined by HPLC were 99.7%, 96.2% and 97.6%, respectively. Their chemical structures were identified by electron spray ionization mass spectroscopy (ESI-MS), 1H, 13C nuclear magnetic resonance (NMR).

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Preparative Separation of Four Major Bufadienolides from the Chinese Traditional Medicine, Chansu, Using High-Speed Counter-Current Chromatography

Natural Product Communications ◽

10.1177/1934578x1000500709 ◽

2010 ◽

Vol 5 (7) ◽

pp. 1934578X1000500 ◽

Cited By ~ 1

Author(s):

Xiu Lan Xin ◽

Junying Liu ◽

Xiao Chi Ma ◽

Qing Wei ◽

Li Lv ◽

...

Keyword(s):

High Speed ◽

Solvent System ◽

Two Phase ◽

Isolation And Purification ◽

H Nmr ◽

Chemical Structures ◽

One Step ◽

Counter Current ◽

Chloroform Methanol ◽

C Nmr

A preparative, high-speed, counter-current chromatographic (HSCCC) method for the isolation and purification of bufadienolides from Chansu was successfully developed by using stepwise elution with a two-phase solvent system composed of n-hexane: chloroform: methanol: water (4:1:2.5:5 and 4:1:4:5, v/v). A total of 7.5 mg of cinobufotalin (1), 8.0 mg of bufalin (2), 14.0 mg of cinobufagin (3) and 9.5 mg of resibufogenin (4) were obtained in a one-step separation from 80 mg of the crude extract with purities of 93.2%, 98.7%, 99.2%, and 99.4%, respectively. The chemical structures were determined from 1H NMR and 13C NMR spectroscopic data.

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Preparative Separation and Purification of Liquiritin and Glycyrrhizic Acid from Glycyrrhiza uralensis Fisch by High-Speed Countercurrent Chromatography

Journal of Chromatographic Science ◽

10.1093/chromsci/bmaa050 ◽

2020 ◽

Vol 58 (9) ◽

pp. 823-830

Author(s):

Hao Wang ◽

Hu Shan ◽

Haitao Lü

Keyword(s):

Crude Extract ◽

High Speed ◽

Glycyrrhizic Acid ◽

Retention Rate ◽

Glycyrrhiza Uralensis ◽

Ionization Mass ◽

Countercurrent Chromatography ◽

Separation And Purification ◽

Isolation And Purification ◽

Glycyrrhiza Uralensis Fisch

Abstract The technique of high-speed countercurrent chromatography (HSCCC) was applied to the preparative isolation and purification of liquiritin and glycyrrhizic acid from a crude extract of Glycyrrhiza uralensis Fisch for the first time. Using single factor and orthogonal design experiments, the best extraction conditions were 70% ethanol, 1:25 ratio of solid-to-liquid (w/v) and extracted 1.5 h at 80°C. The contents of liquiritin and glycyrrhizic acid in the crude extract were 1.3 and 5.3%, respectively. Using the two-phase solvent system of ethyl acetate–methanol–water (5:2:5, v/v), 6.0 mg liquiritin (the purity was 96.7%, and the recovery was 89.3%), and 20.5 mg glycyrrhizic acid (the purity was 98.9%, and the recovery was 77.1%) were obtained from 500 mg crude extraction by HSCCC, respectively. The retention rate of stationary phase was 51.0%. Their structures were identified by high-performance liquid chromatography, melting points, ultraviolet radiation, Fourier transform infrared (FTIR), Electrospray ionization mass spectrometry (ESI-MS), 1H Nuclear Magnetic Resonance (NMR) and 13C NMR spectra. The scavenging abilities of glycyrrhizic acid to 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radicals were stronger than those of liquiritin.

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Isolation and Purification of Aloin a and Aloin B by High-Speed Counter-Current Chromatography

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.634-638.1241 ◽

2013 ◽

Vol 634-638 ◽

pp. 1241-1246 ◽

Cited By ~ 1

Author(s):

Shi Rong Tang ◽

Shang Long Chen ◽

Sang Sang Lu ◽

Xin Yang Hu

Keyword(s):

High Speed ◽

High Performance ◽

Volume Ratio ◽

Solvent System ◽

Two Phase ◽

Lower Phase ◽

Isolation And Purification ◽

Counter Current ◽

Chloroform Methanol ◽

Crude Methanol Extract

High-speed counter-current chromatography(HSCCC) was successfully used for isolation and purification of Aloin A and Aloin B from the crude methanol extract of Aloe with a two-phase solvent system composed of chloroform–methanol–n-butylalcohol-water at an optimized volume ratio of 4:3:1:2 (v/v/v/v). The lower phase was used as the mobile phase in the head to tail elution mode. The preparative HSCCC separation was performed on 180 mg of the crude extract yielding pure Aloin A(18mg) and Aloin B(16mg) at purities of 95.2% and 96.8%, respectively, as determined by high performance liquid chromatography (HPLC). HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.

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Isolation and Purification of Ginkgo Flavonoids by High-Speed Counter-Current Chromatography

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.781-784.741 ◽

2013 ◽

Vol 781-784 ◽

pp. 741-745

Author(s):

Sang Sang Lu ◽

Hui Song ◽

Jing Zhi Miao ◽

Shi Rong Tang

Keyword(s):

Mobile Phase ◽

High Speed ◽

High Performance ◽

Leaf Extract ◽

Volume Ratio ◽

Solvent System ◽

Two Phase ◽

Lower Phase ◽

Isolation And Purification ◽

Counter Current

High-speed counter-current chromatography (HSCCC) was successfully used for isolation and purification of Ginkgo flavonoids from the Ginkgo biloba L. leaf extract (GBE) with a two-phase solvent system composed of n-hexaneethyl acetatemethanolwater at an optimized volume ratio of 4:6:5:5(v/v/v/v). The lower phase was used as the mobile phase in the head to tail elution mode. The preparative HSCCC separation was performed on 200 mg of GBE yielding pure Quercetin (22mg), Kaempferol (15mg) and Isorhamnetin (4mg) at purities of 96.6%, 92.3% and 93.6%, respectively, as determined by high performance liquid chromatography (HPLC). HSCCC is a powerful technique for isolation and separation of chemical composition from GBE.

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