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Method for separating three tannin monomer components from garden burnet

A technology of monomer components and Burnet, which is applied in the field of high-performance liquid chromatography separation, can solve the problems of low yield, time-consuming, easy residue of mobile phase, etc., and achieve fast separation speed, high separation efficiency and good industrial prospects Effect

Active Publication Date: 2014-07-09
CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
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Problems solved by technology

[0010] See the separation method of separating Burnet tannins in the literature, which is mainly column chromatography technology (Wang Manli, Chen Tong. Column chromatography separation of tannins and saponins in Burnet, Chromatography, 1994, 12 (4)), However, this method has low yield, time-consuming and labor-intensive, and can only separate the total tannins and total saponins, but cannot obtain tannin monomer components
So far, there has not been disclosed a method for separating the three monomer components in the crude extract of Burnet burnet by high performance liquid chromatography, which is a separation method that can quickly and efficiently separate the tannin components in the crude extract of Burnet burnet, However, the conditions of the mobile phase should be strictly limited, because most of the tannin components have a phenolic hydroxyl structure, which is prone to ionization and the eluent is easily destroyed during the concentration process and the mobile phase is easy to remain. The choice and its conditions are strictly limited

Method used

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  • Method for separating three tannin monomer components from garden burnet
  • Method for separating three tannin monomer components from garden burnet
  • Method for separating three tannin monomer components from garden burnet

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Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1 Separation and purification method of the present invention

[0044] (1) Pretreatment of raw materials

[0045] A certain amount of crude extract of Burnet burnet was weighed, dissolved in 80% methanol aqueous solution, ultrasonicated for 5 minutes, filtered with a 0.22 μm filter membrane, and set aside.

[0046] Burnet crude extract can be a mixture of a group of tannin compounds extracted from the dried rhizome of Burnet by using the prior art.

[0047]The solvent has a great influence on the separation and stability of the sample. Since the sample has a certain fat solubility and at the same time needs to take into account the hydrophilic compounds, the crude extract of Burnet burnet is stable and has good solubility in methanol aqueous solution. The resolution is also good during analysis, so 80% methanol is selected as the solvent.

[0048] (2) Preparative HPLC separation of three monomer components in Burnet Burnet crude extract

[0049] Take the B...

Embodiment 2

[0063] Embodiment 2: determination of mobile phase

[0064] Several mobile phases with different compositions were selected to separate various monomer components contained in the Burnet extract, among which the mobile phase composed of acetonitrile-hydrochloric acid (pH3. It can achieve good separation of various components, but considering the high price of acetonitrile, we chose relatively economical methanol as the mobile phase under the condition of the same separation effect. Therefore, methanol-hydrochloric acid (pH3.0) aqueous solution was finally selected as the mobile phase system when preparing samples, and in order to obtain better resolution, the mobile phase composed of methanol-phosphoric acid (0.1%) aqueous solution was selected as the mobile phase when detecting samples. Mutually.

[0065] The influence of the pH value of the mobile phase on the separation and stability of each component was investigated. According to the chemical composition characteristics ...

Embodiment 3

[0068] Embodiment 3: Analytical high performance liquid chromatography detects and separates three kinds of monomer components obtained

[0069] When the three components obtained in Example 1 were detected, its liquid chromatography conditions were specifically: chromatographic column: Inertsil, ODS (C 18 ), 5μm, 250mm×4.6mmm ID. Flow rate: 1.0ml / min; Column temperature: 40°C; Detection wavelength: 254nm; Injection volume: 60μl. Mobile phase A: 0.1% phosphoric acid solution; mobile phase B: methanol. The mobile phase gradient is shown in Table 3.

[0070] Table 3 Mobile phase gradient

[0071] time (min)

[0072] See the liquid chromatogram Figure 3-5 , image 3 is the liquid chromatogram of gallic acid components, Figure 4 It is the liquid chromatogram of 3,3′,4′-O-trimethyl retrogallate-4-O-β-D-xyloside component, Figure 5 It is a liquid chromatogram of 3,3',4'-O-trimethyl retrogallic acid components.

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Abstract

The invention provides a method for separating three tannin monomer components from garden burnet. The three tannin monomer components comprises gallic acid, 3,3',4'-O-trimethylellagic acid-4-O-beta-D-xyloside and 3,3',4'-O-trimethylellagic acid. The method utilizes a preparative high performance liquid chromatography to separate and prepare the tannin monomer components, realizes purity more than 98%, has high separation efficiency and a fast separation rate and has a good industrial prospect. The tannin monomer components can be used as tannin standards and can be widely used in research fields of drugs, pharmaceutical analysis and pharmacology.

Description

technical field [0001] The present invention relates to a method for separating three tannin monomer components from Burnet, mainly gallic acid, 3,3',4'-O-trimethyl retrogallate-4-O-β-D-wood High performance liquid chromatographic separation of glycosides and 3,3',4'-O-trimethylretrogallic acid. Background technique [0002] Tannins, also known as tannins, are a class of complex polyphenolic compounds present in plants. In addition to common astringent, antibacterial and anti-inflammatory, hemostatic, anthelmintic, antidiarrheal, anti-diarrheal, anti-infective diseases of various pathogens, its pharmacological activities are also shown in anti-tumor, anti-human immunodeficiency virus (HIV), anti- Mutation, anti-lipid peroxidation, anti-aging, cardiovascular disease, improvement of liver and kidney function, prevention of stress gastrointestinal injury, anti-cataract, detoxification of heavy metal salts and alkaloids have unique effects. [0003] According to the chemical s...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 邹文俊高小平吴建明李小芳代燕平
Owner CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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