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Apigenin on-line two-dimensional composite liquid-phase extraction purification method

A purification method and two-dimensional liquid chromatography technology, which are applied in the field of online two-dimensional composite liquid-phase extraction and purification of apigenin, can solve the problems of low immunity, excessive drug residues, increased secondary infection and secondary infection, etc.

Active Publication Date: 2015-11-11
BEIJING UNIV OF AGRI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, the abuse of antibiotics causes the disorder of normal flora in livestock and poultry, resulting in low immunity, increasing the possibility of secondary infection and secondary infection, making the treatment effect worse and worse, forming a vicious circle
Secondly, a large number of antibiotic drugs used in intensive farming will also cause excessive drug residues, which will seriously affect the import and export trade of animal products and endanger human health.

Method used

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  • Apigenin on-line two-dimensional composite liquid-phase extraction purification method
  • Apigenin on-line two-dimensional composite liquid-phase extraction purification method
  • Apigenin on-line two-dimensional composite liquid-phase extraction purification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Step 1. Accurately weigh 10.0g of parsley leaf powder, add it to a 500ml Erlenmeyer flask, add 100ml of 95% ethanol solution, put the Erlenmeyer flask into an ultrasonic cleaning machine, set the ultrasonic power to 250W, and extract 0.5 at a temperature of 65°C. h. 2.3 Filter with a 0.45μm microporous membrane to separate the extract from the filter residue. Add 70ml of 95% ethanol solution to the filter residue and extract with ultrasound for 0.5h. Repeat 2 times. The three extracts were combined, concentrated to dryness by rotary evaporation, dissolved in a certain volume of methanol, and set aside.

[0119] Step 2. Separation of crude apigenin on-line two-dimensional liquid chromatography

[0120] Weigh 0.4 g of the crude parsley sample and 1.2 g of C4 and mix them into a rotary evaporator, add about 15 mL of methanol, ultrasonically dissolve it completely, and place it in a 40°C rotary evaporator to remove the methanol by rotary evaporation to obtain a dry uniform pow...

Embodiment 2

[0129] Example 2. Method for detecting apigenin content

[0130] HPLC detection

[0131] Sample preparation

[0132] Dissolve 10mg apigenin standard product in a small amount of methanol, place it in a 25mL volumetric flask, shake the methanol to a constant volume, filter it with a 0.22um organic membrane for later use;

[0133] Take the separated fractions of each sample collected in 4.3 and filter them with 0.22um organic membrane for use.

[0134] Chromatographic conditions for detection

[0135] Chromatographic column: C18; column temperature: room temperature; mobile phase: methanol 0.4%, phosphoric acid aqueous solution (1:1); flow rate: 0.8 ml / min; detection wavelength: 346 nm. The samples prepared in 5.1 were detected by high performance liquid chromatography.

[0136] Determination of apigenin content

[0137] The apigenin content was determined by high performance liquid chromatography. Accurately weigh 52.6mg of apigenin reference substance, dissolve it in an appropriate amount...

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Abstract

The invention relates to an apigenin on-line two-dimensional composite liquid-phase extraction purification method which comprises the following steps: 1. taking dry celery leaves, extracting with ethanol, filtering the extracting solution, and concentrating to a dry state; 2. mixing the extract in the step 1 with a C4 filler, adding methanol, and carrying out rotary evaporation in a 40-DEG C rotary evaporator to remove methanol, thereby obtaining dry powder; and 3. filling a preparative chromatographic column with the dry powder obtained in the step 2, connecting into an on-line two-dimensional liquid chromatography separation system, and carrying out separation.

Description

Technical field: [0001] The invention relates to a method for extracting and separating components in vegetables, in particular to an online two-dimensional composite liquid phase extraction and purification method for apigenin. Background technique: [0002] With the advancement and development of the animal husbandry and veterinary industry, in some respects, we have discovered more diseases, more bacteria and viruses, and many mutations have taken place in the existing bacteria and viruses. Large-scale intensive breeding of livestock and poultry increases the possibility of animal disease, increases the occurrence of infectious diseases, and reduces the resistance of animals to general pathogens. In order to resist the invasion of pathogens, treat diseases, ensure production, and reduce profit loss, people must increase the amount and types of drugs used, such as antibiotics. However, the extensive use of antibiotics will also bring about many serious problems, which are...

Claims

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Application Information

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IPC IPC(8): C07D311/30C07D311/40
CPCC07D311/30C07D311/40
Inventor 崔德凤张永红王建舫李佳沈红
Owner BEIJING UNIV OF AGRI