Three-phase solvent systems for the comprehensive separation of a wide variety of compounds fromDicranostigma leptopodumby high-speed counter-current chromatography

In order to obtain high-purity flavonoid products, the extracts from mulberry leaves were separated and purified via high-speed counter-current chromatography (HSCCC). Moreover, the product was detected via high-performance liquid chromatography (HPLC). The characteristic absorption wavelength of the rutin standard for HSCCC detection and HPLC analysis at 257 nm was tested by ultraviolet scanning analysis. The effect of solvent systems and mobile phase flow rate on the separation efficiency were then researched. Finally, the solvent system of V(ethyl acetate):V(n-butanol):V(water) = 4:1:5 was selected as the operating system for HSCCC. This work theoretically analyzed the impact of the molecular structure and polarity of flavonoids on the choice of solvent systems. The results showed that the mobile phase flow rate had a great influence on the separation efficiency. Furthermore, the separation efficiency increased as the mobile phase flow rate decreased. When the mobile phase flow rate was 5 mL/min, the peak time for flavonoids was 140 min, the retention of the stationary phase was 56.4%, and the purity of the product reached 93.8%. The results of this study greatly improved the purity of flavonoids in mulberry leaf and provided a strong support for the separation and purification of mulberry leaf extract.

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A Three-Phase Solvent System in High-Speed Counter-Current Chromatographic for the Separation and Purification of Bioactive Constituents from Acanthus ilicifolius

Chromatographia ◽

10.1007/s10337-015-2952-5 ◽

2015 ◽

Vol 78 (21-22) ◽

pp. 1401-1407 ◽

Cited By ~ 1

Author(s):

Fazuo Wang ◽

Ru Li ◽

Lijuan Long ◽

Xinpeng Tian ◽

Zhihui Xiao ◽

...

Keyword(s):

High Speed ◽

Solvent System ◽

Separation And Purification ◽

Bioactive Constituents ◽

Acanthus Ilicifolius ◽

Three Phase ◽

Counter Current

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Efficient Preparation of Bafilomycin A1 from Marine Streptomyces lohii Fermentation Using Three-Phase Extraction and High-Speed Counter-Current Chromatography

Marine Drugs ◽

10.3390/md18060332 ◽

2020 ◽

Vol 18 (6) ◽

pp. 332

Author(s):

Ye Yuan ◽

Xiaoping He ◽

Tingting Wang ◽

Xingwang Zhang ◽

Zhong Li ◽

...

Keyword(s):

Crude Extract ◽

High Speed ◽

Solvent System ◽

Bafilomycin A1 ◽

Middle Phase ◽

Two Phase ◽

Rapid Separation ◽

Lower Phase ◽

Three Phase ◽

Counter Current

An efficient strategy was developed for the rapid separation and enrichment of bafilomycin A1 (baf A1) from a crude extract of the marine microorganism Streptomyces lohii fermentation. This strategy comprises liquid−liquid extraction (LLE) with a three-phase solvent system (n-hexane–ethyl acetate–acetonitrile–water = 7:3:5:5, v/v/v/v) followed by separation using high-speed counter-current chromatography (HSCCC). The results showed that a 480.2-mg fraction of baf A1-enriched extract in the middle phase of the three-phase solvent system was prepared from 4.9 g of crude extract after two consecutive one-step operations. Over 99% of soybean oil, the main hydrophobic waste in the crude extract, and the majority of hydrophilic impurities were distributed in the upper and lower phase, respectively. HSCCC was used with a two-phase solvent system composed of n-hexane–acetonitrile–water (15:8:12, v/v/v) to isolate and purify baf A1 from the middle phase fraction, which yielded 77.4 mg of baf A1 with > 95% purity within 90 min. The overall recovery of baf A1 in the process was determined to be 95.7%. The use of a three-phase solvent system represents a novel strategy for the simultaneous removal of hydrophobic oil and hydrophilic impurities from a microbial fermentation extract.

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