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Method for separating and purifying cordycepin by utilizing high-speed counter-current chromatography

A high-speed countercurrent chromatography, separation and purification technology is applied in the field of high-speed countercurrent chromatography to separate and purify high-purity cordycepin, to achieve the effects of high cordycepin yield and purity, good peak separation effect, and simple operation method.

Active Publication Date: 2012-01-18
ZHONGSHAN LEAGUE PRODS
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, no one has used high-speed countercurrent chromatography to separate and purify cordycepin

Method used

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  • Method for separating and purifying cordycepin by utilizing high-speed counter-current chromatography
  • Method for separating and purifying cordycepin by utilizing high-speed counter-current chromatography
  • Method for separating and purifying cordycepin by utilizing high-speed counter-current chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] The practice and drugs used in the experiment are shown in Table 1

[0074] Table 1 Experimental drugs and reagents

[0075]

[0076] 1. Sample pretreatment

[0077] Take 900ml of the crude cordycepin extract, and centrifuge for 20min at a speed of 3500r / min. About 800ml of supernatant. Seal the collected supernatant and store it at low temperature for future use.

[0078] The content of cordycepin in the supernatant of the crude extract of cordycepin was determined by high performance liquid chromatography, the chromatographic conditions were: mobile phase: methanol: water=15:85 (V / V), detection wavelength: 260nm, Column temperature: room temperature, flow rate: 1ml / min, injection volume: 20μl, t R About 10min. Its chromatogram see figure 2 .

[0079]We can see from the chromatogram that the substance in the crude extract of cordycepin is very impurity, especially there are many impurity peaks in the front part of the high-performance liquid chromatogram, bu...

Embodiment 2

[0160] The difference from Example 1 is that the two-phase solvent system for isolating cordycepin is n-hexane: n-butanol: methanol: water=25:75:25:150, K=0.5, and the obtained mass percentage is 98.79 % pure cordycepin ( Figure 16 , t=9.81).

Embodiment 3

[0162] The difference from Example 1 is that the two-phase solvent system for separating cordycepin is n-hexane: n-butanol: methanol: water=25:70:28:145 (V / V), K=1.0, and the mass 99.85% pure cordycepin ( Figure 17 , t=9.79).

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Abstract

The invention discloses a method for separating and purifying cordycepin by utilizing a high-speed counter-current chromatography. The method is characterized by comprising the following steps: 1, preparing a cordycepin sample solution; and 2, preparing a two-phase solvent system which can be used for the high-speed counter-current chromatography and has a two-phase distribution coefficient K value ranging from 0.5 to 1.0 according to the physicochemical property of the cordycepin, then carrying out high-speed counter-current chromatography separation and purification on the cordycepin sample solution obtained in the step 1 by using the two-phase solvent system, and collecting a flowing-out mobile phase solution or immobile phase solution, so as to obtain a cordycepin solution pure product. In the invention, the two-phase solvent system having the two-phase distribution coefficient K value ranging from 0.5 to1.0 has excellent good separation effect on the cordycepin, and the peak separation effect in the high-speed counter-current chromatography is extremely good, thus the purity of the cordycepin pure product obtained by separation is extremely high and can reach above 98%.

Description

technical field [0001] The invention relates to a method for separating and purifying cordycepin, in particular to a method for separating and purifying high-purity cordycepin by using high-speed countercurrent chromatography. Background technique [0002] Cordycepin is a characteristic characteristic component of fungi of the genus Cordyceps, which generally does not contain this component in other edible and medicinal fungi. Therefore, this component has special significance for identifying Cordyceps products. Cordycepin (3′-deoxyadenosine) is a nucleoside antibiotic belonging to purine alkaloids with the molecular formula C 10 h 13 N 5 o 3 , see structural formula I for the structural formula, molecular weight 251, soluble in water, hot ethanol, methanol, n-butanol and n-propanol, insoluble in benzene, butyl ether and chloroform, melting point 225-226°C, optical rotation -47°. [0003] [0004] The content of cordycepin in fungi of the genus Cordyceps is very low,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H19/16C07H1/06
Inventor 傅少杰高素萍李光仪
Owner ZHONGSHAN LEAGUE PRODS
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