ABSTRACT
This study aimed at preparing the chemical reference substance of garcinol.A preparative high-speed countercurrent chromatography (HSCCC) was adopted for the isolation of garcinol from the twigs of Garcinia multifiora Champ,G.esculenta Y.H.Li and the fruits of G.xanthochymus Hook.f.ex T.Anders.The crude extracts were separated by HSCCC with two phase solvent systems composed of n-hexane-ethyl acetate-95％ ethanol-water (8∶8∶12∶4,volume ratio) using the upper stationary phase and the lower mobile phase.Under the conditions of a flow rate of 2.0 and 4.0 mL· min-1,and the apparatus rotation at 850 rpm,the column temperature at 25℃ and the detection wavelength at 254 nm,the fraction containing garcinol was purified by the Sephadex LH20 chromatography.The purity test in the extracts was determined by high-performance liquid chromatography.As a result,the contents of garcinol were 45,281 and 4080mg respectively separated from 104.765 g twigs of G.multiflora Champ,105.270 g G.esculenta Y.H.Li and 102.318 g the fruits of G.xanthochymus Hook.f.ex T.with the purity of 98.72％,98.36％ and 98.42％.Compared with the sample pretreatments and separation efficiency of the three plants,it was found that the fat-soluble extracts rapidly and efficiently extracted using n-hexane-ethyl acetate-95％ ethanol-water (5∶5∶5∶5,volume ratio) contained abundant garcinol taking fruits of G.xanthochymus Hook.f.ex T as the raw material with the combination of HSCCC and Sephadex LH-20 chromatography.In conclusion,it was indicated that the combination method is efficient with high operability and large preparation quantity.
ABSTRACT
Objective: To establish a simple and effective method for the isolation of leonurine from Leonurus japonicus by high speed countercurrent chromatography (HSCCC). Methods: The extraction conditions of leonurine were optimized by one-factor experimental design. After comparing several different solvent systems, the two phase system of ethyl acetate-n-butanol-water (3∶2∶5) was finally chosen as operating solvent of HSCCC for the separation of leonurine, in which the lower phase was determined as the mobile phase and the upper phase as stationary. The detection of eluates was performed with an ultraviolet detector at 277 nm. The rotation speed was adjusted at 850 r/min, and the flow rate was 2.2 mL/min. Results: Leonurine was successively isolated from n-butanol fraction by HSCCC, and the above established method was also successively applied to the crude extact of L. japonicus. Finally, 68 mg of leonurine with purity about 96.2% could be obtained from 2.48 g crude extract of L. japonicus in a single injection. Conclusion: The described approaches actively promote efficient preparation strategy to obtain high purity of leonurine.
ABSTRACT
Extraction and separation technologies appeared in recent years are particularly reviewed in the principles, features, latest research and application results, and existing problems as well. These new techniques include ultrahigh pressure extraction (UPE), ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), pressurized solvent extraction (PSE), smashing tissue extraction (STE), heating-free extraction (HFE), high-speed countercurrent chromatography (HSCCC), and molecular imprinting technique (MIT). Further research topics and application prospects of these extraction and separation technologies are also suggested, so as to provide the reference for the research and production of modern Chinese materia medica.
ABSTRACT
Objective: To separate and purify coumarins from Heracleum millefolium by high-speed counter-current chromatography (HSCCC). Methods: HPLC was used to analyze and optimize the solvent system and hexane-ethyl acetate-water-methanol (1.5:2.5:2:1.5) was chosen as the two-phase solvent system of HSCCC, in which the upper phase was used as the stationary phase, while the lower phase was used as the mobile phase with flow rate of 3 mL/min, the revolution speed was set at 850 r/min, and detected at 323 nm. Three compounds were obtained as crystals by vacuum concentration at 50℃, and determined through HPLC, then their structures were identified by IR, ESI-MS, 1H-NMR, and 13C-NMR. Results: Columbianetin acetate (6.3 mg), osthole (10.6 mg), and columbianadin (6.4 mg) were obtained from crude sample (300 mg) extracts and their purity were above 96.0%. Conclusion: HSCCC is a powerful technique for the rapid separation and purification of coumarins from crude extract of H. millefolium.
ABSTRACT
Objective: To develop an efficient method for separating and purifying puerarin from the roots of Pueraria lobata. Methods: Separation by fast centrifugal partition chromatography (FCPC) was processed with a biphasic solvent system composed of ethyl acetate- n-butanol-water (2:1:3). The separation conditions were determined as follows: sample loading of 10 mg, flow rate of 2 mL/min, rotation speed of 2200 r/min, ascending mode, and detection wavelength of 254 nm. High speed countercurrent chromatography (HSCCC) was used as a comparative method with the rotation speed of 800 r/min, flow rate of 2.0 mL/min, and sample loading of 10 mg in tail-to-head mode. Results: Puerarin was obtained by FCPC with a resolution of 0.90 and a purity above 99%, while a resolution below 0.50 and a purity below 90% by HSCCC. Compared with HSCCC, FCPC has the advantages with higher purity and better resolution. Conclusion: FCPC is a powerful method to separate and purify puerarin. © 2014 Tianjin Press of Chinese Herbal Medicines.
ABSTRACT
An efficient method for the isolation and purification of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high speed counter-current chromatography (HSCCC) was established in this paper.The ether extracts of Radix Eupatorii Chinensis were purified by HSCCC with a solvent system of hexyl hydride ethyl acetate-methanol-water (1∶2∶1∶2,v/v/v/v).The upper phase was used as the stationary phase and the lower phase as the mobile phase.About 8.4 mg of 12,13-dihydroxyeuparin was obtained from 200 mg of ether extracts from Radix Eupatorii Chinensis in one-step HSCCC separation,with the purity of 96.71％,as determined by HPLC.After methanolwater recrystallization,the purity of 12,13-dihydroxyeuparin reached 99.83％.Such a simple and effective method was fairly useful to prepare pure compound as reference substances for related study on Radix Eupatorii Chinensis.
ABSTRACT
Objective: To introduce a new way, which facilitates the systematic search and evaluation of the solvent systems suitable for high-speed countercurrent chromatography (HSCCC). Methods: The software integrated with the thermodynamics solution functions and theory is directive to calculate the composition, physical parameters, such as volume, viscosity, dielectric constants, and polarity parameters of the upper and lower phases. Results: Separation of Guizhi-decoction Fr. A by HSCCC, one of traditional Chinese medcines, is conducted to prove the applicability of this software in selection and optimization of solvent systems. Conclusion: This software can be used to select and optimize the solvent systems of HSCCC. It is supposed to extend this method.