CN111214505A - Heavy metal removal process for ginkgo biloba extract - Google Patents

Abstract

The invention discloses a process for removing heavy metals from a ginkgo biloba extract, which comprises the steps of extracting, performing column chromatography, adsorbing by a nano material, separating by the nano material, concentrating and drying a separation solution to obtain the ginkgo biloba extract; the nano material is a ferroferric oxide nano material, and the nano material separation technology is a magnetic separation technology. As a refinement, the method may further comprise a nanomaterial recovery step. The invention can utilize Fe₃O₄Adsorbing ginkgolic acid, urushiol, isourushiol, and heavy metals with nanometer material, performing solid-liquid separation by magnetic separation, concentrating medicinal liquid, and drying to obtain the final product. The method is environment-friendly, and has double effects of removing ginkgolic acid and heavy metal.

Description

Heavy metal removal process for ginkgo biloba extract

Technical Field

The invention belongs to the technical field of natural product preparation, and particularly relates to a method for removing ginkgolic acid and heavy metals in a ginkgo leaf extract by utilizing the adsorption effect of a nano material.

Background

Folium Ginkgo is dry leaf of Ginkgo biloba (Ginkgobilobal.) of Ginkgoliaceae, has sweet, bitter, astringent and mild properties, and has effects of astringing lung, relieving asthma, promoting blood circulation for removing blood stasis, and relieving pain, and can be used for treating coronary heart disease, angina pectoris and hyperlipidemia. The folium Ginkgo extract is prepared from folium Ginkgo, and has effects in promoting blood circulation, dispelling blood stasis, and dredging collaterals, and can be used for treating thoracic obstruction, cardialgia, apoplexy, hemiplegia, and stiff tongue caused by blood stasis and obstruction of collaterals; coronary heart disease stable angina pectoris, cerebral infarction and the above syndromes. Folium ginkgo and folium ginkgo extracts are used as raw materials, and are widely used for preparing preparations for treating cardiovascular and cerebrovascular diseases, only Chinese pharmacopoeia records 5 related varieties including folium ginkgo, European pharmacopoeia specifies the standards of the folium ginkgo and the folium ginkgo extracts, and United states pharmacopoeia records the folium ginkgo and the preparations thereof as food supplements. However, the ginkgo preparation also contains a toxic component, namely ginkgolic acid, which has the effects of sensitization, cytotoxicity and the like, so the international standard requires that the content of ginkgolic acid is less than 5ppm and the content of heavy metal is less than 10 ppm. The content of ginkgolic acid and heavy metal in the ginkgo preparation produced currently in China greatly exceeds the international standard. Most of ginkgo biloba extract production enterprises in China are in a production halt state or a semi-production halt state, and how to effectively remove ginkgolic acid and heavy metal in the ginkgo biloba extract is a main problem of the research of chemical workers in China at present.

At present, the methods for removing ginkgolic acid include: (1) acetone extraction + extraction as proposed by Schwabe corporation; (2) acetone extraction, chromatography and organic solvent extraction; (3) ethanol extraction, chromatography and organic solvent extraction; (4) and (4) performing supercritical extraction. The solvent extraction is easy to produce emulsification, so that the production time is prolonged, the extraction is incomplete, the solvent residue exists in the product, and the method is not friendly to operators and environment. The patent document "process for removing ginkgolic acid from folium Ginkgo extract" (CN101194918A) adopts a double-column stacking method to remove ginkgolic acid, but the manufacturing process is relatively complicated, and the pretreatment and activation of the double columns can generate a certain amount of industrial wastewater.

The excessive ginkgolic acid and heavy metal content in the ginkgo leaf extract is a bottleneck for restricting the export of the ginkgo leaf extract in China, and simultaneously, the pharmaceutical safety in China is also influenced.

Disclosure of Invention

The invention aims to provide a method for removing ginkgolic acid and heavy metals in a ginkgo leaf extract by using a nano material, which has the advantages of environmental friendliness and double effects of removing ginkgolic acid and heavy metals.

The invention provides a method for removing ginkgolic acid and heavy metal in a ginkgo leaf extract by utilizing a nano material, which is characterized in that the ginkgo leaf extract is obtained by extracting, column chromatography, nano material adsorption, nano material separation, and concentration and drying of a separation solution; the nano material is a ferroferric oxide nano material, and the nano material separation technology is a magnetic separation technology.

As an improvement of the above technical scheme, the method also comprises a nanometer material recovery step.

As another improvement of the above technical solution, the extraction includes the following specific processes: adding 60-70% ethanol into folium Ginkgo respectively, reflux-extracting folium Ginkgo and ethanol solution at a mass volume ratio of 1: 5-1: 10 for at least 2 times, each for 1-3 hr, mixing extractive solutions, recovering ethanol under reduced pressure, adding 3-5 times of water, standing, filtering, and collecting filtrate.

As a further improvement of the above technical scheme, the column chromatography comprises the following specific processes: loading any one of 4 kinds of activated resins AB-8, HP-20, DA201, and D101 into column, washing the filtrate obtained in the extraction step with 3-6 times of resin bed volume, desorbing with 2-4 times of resin bed volume of 60-80% ethanol, and collecting ethanol eluate.

As a further improvement of the above technical solution, the nanomaterial adsorbs the following specific processes: adding a ferroferric oxide nano material into the collected ethanol eluent, wherein the mass ratio of the ferroferric oxide to the volume ratio of the ethanol eluent is 0.01-1%, and performing ultrasonic adsorption for 0.5-8 h.

As a further improvement of the above technical solution, the manner of concentrating and drying the separated liquid is vacuum drying or spray drying.

As a further improvement of the above technical solution, the recovery process of the nanomaterial comprises: collecting the ferroferric oxide nano material obtained in the nano material separation step, soaking the ferroferric oxide nano material in 0.05-2% hydrochloric acid for 0.5-24h, washing the ferroferric oxide nano material with deionized water for a plurality of times, and drying the ferroferric oxide nano material.

Fe₃O₄The nano material has higher affinity with catechol and derivatives thereof, so that the nano material selectively adsorbs some plant chemical substances with carboxyl or ortho-phenolic hydroxyl. The total ginkgolic acid includes ginkgolic acid, isoginkgolic acid, ginkgolic acid, etc., and its basic structure has carboxyl or o-phenolic hydroxyl, Fe₃O₄The nanometer material can adsorb ginkgolic acid, urushiol, iso-urushiol, etc., and magnetic separation technology is used to separate total ginkgolic acid from mother liquor. The invention has the advantages of utilizing Fe₃O₄The nano material not only canTo remove ginkgolic acid and heavy metals, and to obtain the effect of killing two birds with one stone, and Fe₃O₄The nano material is recycled by acid washing, and the enriched ginkgolic acid can be produced into the biological insecticide and other uses in an industrialized mass production.

Detailed Description

The method for removing ginkgolic acid and heavy metal in ginkgo leaf extract by using nano material provided by the specific embodiment of the invention is specifically completed by the following steps:

step 1, extraction: adding 60-70% ethanol into folium Ginkgo respectively, reflux-extracting folium Ginkgo with ethanol solution at a ratio of 5-10(W/V) for at least 2 times, each time for 1-3 hr, mixing extractive solutions, recovering ethanol under reduced pressure, adding 3-5 times of water, standing, filtering, and collecting filtrate.

Step 2, column chromatography: loading activated resin selected from 4 resins such as AB-8, HP-20, DA201, and D101 into column, loading the filtrate into column, washing the column with 3-6 times of water, desorbing with 2-4 times of resin bed volume of 60-80% ethanol, and collecting ethanol eluate.

And 3, adsorbing the nano material: adding 0.01-1% (W/V) ferroferric oxide nano material into the collected ethanol eluent, and performing ultrasonic adsorption for 0.5-8 h.

And 4, separating nano materials: and adsorbing the ferroferric oxide nano material by adopting permanent magnetism or electromagnetism to complete solid-liquid separation.

And 5, concentrating and drying the separated liquid: vacuum drying or spray drying.

And 6, recovering the nano materials: collecting the ferroferric oxide nano material obtained in the nano material separation step, soaking the ferroferric oxide nano material in 0.05-2% hydrochloric acid for 0.5-24h, washing the ferroferric oxide nano material with deionized water for a plurality of times, and drying the ferroferric oxide nano material.

The following examples are provided to further illustrate embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

Example 1

Taking 500g of ginkgo leaves, adding 10-8 times of 60% ethanol, respectively performing reflux extraction for 2h, filtering to obtain liquid medicines, merging, recovering ethanol under reduced pressure until the liquid medicines do not contain alcohol, then adding water to 1500mL, standing, filtering, adding the filtrate to a treated DA201 resin column, performing column cleaning by purified water, eluting by 2000mL of 80% ethanol when the effluent liquid is clear, collecting ethanol eluent, adding 0.2g of ferroferric oxide nano material, performing ultrasonic adsorption for 2h, putting a magnet into the liquid medicines to adsorb the ferroferric oxide nano material, performing reduced pressure concentration on the liquid medicines to recover ethanol, and performing vacuum drying on the extract. Collecting ferroferric oxide nano material, soaking in 0.1% hydrochloric acid for 4h, magnetically separating, washing with deionized water for several times, and vacuum drying for use. Weighing the dried ginkgo leaf extract, sampling and determining the content of total flavonoids, total lactones, ginkgolic acids and heavy metals. The results were: 9.1g of ginkgo leaf extract, 28.3 percent of total flavone content, 8.6 percent of total lactone, 2ppm of ginkgoic acid and 4ppm of heavy metal.

Example 2

Taking 500g of ginkgo leaves, adding 70% ethanol of which the amount is 8 to 6 times that of the ginkgo leaves, respectively carrying out reflux extraction for 1.5h, filtering to obtain liquid medicine, merging, recovering ethanol under reduced pressure until the liquid medicine does not contain alcohol, then adding water to 1500mL, standing, filtering, adding filtrate to a treated D101 resin column, carrying out purified water column chromatography, eluting with 2500mL 80% ethanol when the effluent is clear, collecting ethanol eluate, recovering ethanol under reduced pressure until the solid content of the liquid medicine reaches 1.08-1.10, stopping concentration, adding 0.5g of ferroferric oxide nano material into the liquid medicine, carrying out ultrasonic adsorption for 5h, putting an electromagnet into the liquid medicine to adsorb the ferroferric oxide nano material, carrying out reduced pressure concentration on the liquid medicine, and carrying out spray drying when the solid content reaches 1.25-1.30. Collecting the ferroferric oxide nano material, soaking in 1% hydrochloric acid for 2h, performing magnetic separation, washing with deionized water for several times, and vacuum drying for later use. Weighing the dried ginkgo leaf extract, sampling and determining the content of total flavonoids, total lactones, ginkgolic acids and heavy metals. The results were: 10.2g of ginkgo leaf extract, 27.2 percent of total flavone content, 8.2 percent of total lactone, 2ppm of ginkgoic acid and 5ppm of heavy metal.

Example 3

Taking 500g of ginkgo leaves, adding 70% ethanol in an amount which is 10 and 5 times that of the ginkgo leaves, respectively carrying out reflux extraction for 3 hours, filtering to obtain liquid medicine, merging, recovering ethanol under reduced pressure until the liquid medicine does not contain alcohol, then adding water to 1500mL, standing, filtering, adding the filtrate to a treated AB-8 resin column, carrying out column cleaning by purified water, eluting by 2000mL of 60% ethanol when the effluent is clear, collecting ethanol eluate, adding 1.0g of ferroferric oxide nano material, carrying out ultrasonic adsorption for 8 hours, putting a magnet into the liquid medicine to adsorb the ferroferric oxide nano material, carrying out reduced pressure concentration on the liquid medicine, and carrying out spray drying when the solid content reaches 1.25-1.30. Collecting the ferroferric oxide nano material, soaking in 2% hydrochloric acid for 24h, carrying out magnetic separation, washing with deionized water for several times, and carrying out vacuum drying for later use. Weighing the dried ginkgo leaf extract, sampling and determining the content of total flavonoids, total lactones, ginkgolic acids and heavy metals. The results were: 8.8g of ginkgo leaf extract, 26.6 percent of total flavone content, 7.4 percent of total lactone, 1ppm of ginkgoic acid and 3ppm of heavy metal.

Example 4

Taking 500g of ginkgo leaves, adding 10-8 times of 65% ethanol, respectively carrying out reflux extraction for 2h, filtering to obtain liquid medicine, merging, recovering ethanol under reduced pressure until the liquid medicine does not contain alcohol, then adding water to 1500mL, standing, filtering, adding the filtrate to a treated HP-20 resin column, carrying out purified water column filtration, eluting with 2800 mL 80% ethanol when the effluent is clear, collecting ethanol eluate, recovering ethanol under reduced pressure until the solid content of the liquid medicine reaches 1.08-1.10, stopping concentration, adding 5.0g of ferroferric oxide nano material, carrying out ultrasonic adsorption for 3h, putting a magnet into the liquid medicine to adsorb the ferroferric oxide nano material, carrying out reduced pressure concentration on the liquid medicine, and carrying out spray drying when the solid content reaches 1.25-1.30. Collecting ferroferric oxide nano material, soaking in 2% hydrochloric acid for 3h, magnetically separating, washing with deionized water for several times, and vacuum drying. Weighing the dried ginkgo leaf extract, sampling and determining the content of total flavone, total lactone, ginkgoic acid and heavy metal. The results were: 9.5g of ginkgo leaf extract, 28.6 percent of total flavone content, 7.6 percent of total lactone, 1ppm of ginkgoic acid and 2ppm of heavy metal.

In the present invention, the concentrations of ethanol and hydrochloric acid are both volume percentage concentrations.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention, and the present invention should not be limited by the disclosure of the preferred embodiments. Therefore, it is intended that all equivalents and modifications which do not depart from the spirit of the invention disclosed herein are deemed to be within the scope of the invention.

Claims (1)

1. A process for removing heavy metals from a ginkgo extract is characterized by comprising the following steps:

step 1, extraction: respectively adding 60-70% ethanol into folium Ginkgo, wherein the ratio of folium Ginkgo to ethanol solution is 5W/V-10W/V, reflux extracting for at least 2 times, each time for 1-3 hr, mixing extractive solutions, recovering ethanol under reduced pressure, adding 3-5 times of water, standing, filtering, and collecting filtrate;

step 2, column chromatography: loading any one of 4 kinds of activated resins of AB-8, HP-20, DA201 and D101 into a column, loading the filtrate obtained by extraction into the column, washing the column with 3-6 times of volume of water, desorbing with 60% -80% ethanol 2-4 times of the volume of a resin bed, and collecting the ethanol eluate;

and 3, adsorbing the nano material: adding 0.01-1W/V% ferric oxide nano material into the collected ethanol eluent, performing ultrasonic adsorption for 0.5-8h, and removing ginkgolic acid and heavy metal simultaneously in the process;

and 4, separating nano materials: adsorbing the ferroferric oxide nano material by adopting permanent magnet or electromagnetism to complete solid-liquid separation;

and 5, concentrating and drying the separated liquid: vacuum drying or spray drying;

and 6, recovering the nano materials: collecting the ferroferric oxide nano material obtained in the nano material separation step, soaking the ferroferric oxide nano material in 0.05-2% hydrochloric acid for 0.5-24h, washing the ferroferric oxide nano material with deionized water for a plurality of times, and drying the ferroferric oxide nano material.