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

Marine Drugs ◽  
2020 ◽  
Vol 18 (6) ◽  
pp. 332
Author(s):  
Ye Yuan ◽  
Xiaoping He ◽  
Tingting Wang ◽  
Xingwang Zhang ◽  
Zhong Li ◽  
...  

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.

2013 ◽  
Vol 634-638 ◽  
pp. 1241-1246 ◽  
Author(s):  
Shi Rong Tang ◽  
Shang Long Chen ◽  
Sang Sang Lu ◽  
Xin Yang Hu

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.


2013 ◽  
Vol 781-784 ◽  
pp. 741-745
Author(s):  
Sang Sang Lu ◽  
Hui Song ◽  
Jing Zhi Miao ◽  
Shi Rong Tang

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.


Molecules ◽  
2019 ◽  
Vol 24 (1) ◽  
pp. 197 ◽  
Author(s):  
Ran Yang ◽  
Lei Fang ◽  
Jia Li ◽  
Zhenhua Zhao ◽  
Hua Zhang ◽  
...  

A high-speed counter-current chromatography (HSCCC) method, using a two-phase solvent system composed of ethyl acetate/n-butanol/methanol/water (5:1:1:5, v/v/v/v), was successfully established to separate the five iridoid glucosides 7-O-ethyl sweroside (1), secologanin dimethylacetal (2), adinoside F (3), (7R)-secologain n-butyl methyl acetal (4) and adinoside G (5) from Lonicerae Japonicae Flos. Their purities were 96.8%, 98.5%, 93.3%, 98.0% and 99.9%, respectively. All the iridoid glucosides were identified by HR-ESI-MS, 1D and 2D NMR. Compounds 3 and 5 are new iridoid glucosides. The anti-inflammatory tests showed that compounds 1–5 all expressed moderate inhibitory effects on β-glucuronidase release in rat polymorphonuclear leukocytes (PMNs) induced by platelet-activating factor (PAF) with IC50 values ranging from 4.52 to 6.50 µM, while the antibacterial assays demonstrated that all the compounds displayed mild inhibitory activities against Staphylococcus aureus ATCC 25923 with MIC values ranging from 13.7 to 26.0 µg/mL.


2013 ◽  
Vol 634-638 ◽  
pp. 1502-1505
Author(s):  
Bin Li ◽  
Xian Jun Meng ◽  
Li Jie Zhu ◽  
Xin Yao Jiao

High-speed counter-current chromatography (HSCCC) was successfully applied to the preparative separation and purification of lancifodilactone C from the crude extracts of Schisandra chinensis (Turcz.) Baill. Following an initial cleaning-up step on the AB-8 macroporous resin, a preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of Chloroform- n-Butyl alcohol- methanol -water(10:0.5:8:4,v/v) was used to isolate and separate lancifodilactone C from Schisandra Chinensis(Turcz.) Baill. A total of 101 mg lancifodilactone C with purities of 98.2% were obtained from 1000 mg crude extract in one-step elution and less than 1 h, and the structure identification was performed by UV, IR, MS,1H NMR and13C NMR.


2001 ◽  
Vol 927 (1-2) ◽  
pp. 91-96 ◽  
Author(s):  
L.M Yuan ◽  
M Zi ◽  
P Ai ◽  
X.X Chen ◽  
Z.Y Li ◽  
...  

2015 ◽  
Vol 21 (2) ◽  
pp. 331-341 ◽  
Author(s):  
Zhuoni Hou ◽  
Xianrui Liang ◽  
Feng Su ◽  
Weike Su

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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