A method for extracting quebracho alcohol from seabuckthorn leaves

Abstract

The invention relates to the extraction of quebracho alcohol, and more specifically, to a method for extracting quebracho alcohol from seabuckthorn leaves, comprising the following steps: ultrafine grinding of seabuckthorn leaves, adding pure water to extract at room temperature, centrifuging the filtrate by graded centrifugation, pressing and drying the filter residue to obtain an extract; adsorbing through a macroporous adsorption resin; purifying through a secondary organic solvent nanofiltration membrane to obtain a decolorized intercepted membrane liquid; the decolorized intercepted membrane liquid enters an anion exchange resin, is eluted with pure water, and then enters a cation exchange resin and is eluted with pure water; then passes through a reverse osmosis membrane to obtain an intercepted liquid; the intercepted liquid is concentrated into an extract, and then an organic solvent is added to dissolve, the system is adjusted to be acidic, and then cooled and crystallized, and dried to obtain a quebracho alcohol product. The quebracho alcohol content extracted and prepared by the invention is as high as 97%, and the recovery rate is as high as 80%. The extraction method is safe, environmentally friendly, and simple to operate, and is suitable for large-scale industrial production.

Description

Technical Field

[0001] The invention relates to the extraction of quercetin, and more particularly to a method for extracting quercetin from seabuckthorn leaves. Background Art

[0002] Seabuckthorn is a deciduous shrub of the Elaeagnaceae family and the genus Seabuckthorn. It is drought-resistant and wind-resistant, and can survive on salinized land, so it is widely used in soil and water conservation. The roots, stems, leaves, flowers and fruits of seabuckthorn can be widely used in many fields of the national economy, such as food, medicine, light industry, aerospace, agriculture, animal husbandry and fishery.

[0003] At present, the application and development of seabuckthorn is generally focused on seabuckthorn fruit, while a small part of seabuckthorn leaves, an important resource, is used in seabuckthorn tea, and the remaining large amount is abandoned as waste, resulting in a huge waste of resources. As a methyl ether derivative of inositol, quesilanol has important potential hypoglycemic activity and is a natural active ingredient with great utilization value. It exists in different parts of seabuckthorn plants. Research data confirms that quesilanol is rich in seabuckthorn leaves.

[0004] Since quebrachoa alcohol is a small molecule substance that is easily soluble in water but not easily soluble in organic solvents, it is difficult to obtain a high-content quebrachoa alcohol product. There are no reports in China of large-scale production of quebrachoa alcohol with a content of more than 95%. The reason is that the extraction process is complex, the operation is difficult, and the key production points are difficult to control. Summary of the invention

[0005] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for extracting quesol from seabuckthorn leaves. The quesol content extracted by the method of the present invention is as high as 97%, the recovery rate is as high as 80%, and the extraction method is safe, environmentally friendly, simple to operate, and suitable for large-scale industrial production.

[0006] The above technical purpose of the present invention is achieved through the following technical scheme: a method for extracting quebracho alcohol from seabuckthorn leaves, comprising the following steps:

[0007] (1) Ultrafine grinding of seabuckthorn leaves, adding pure water and stirring, extracting at room temperature, filtering the extracted seabuckthorn leaves through filter cloth, fractionating and centrifuging the filtrate, pressing and drying the filter residue to obtain an extract;

[0008] (2) the extract is passed through a macroporous adsorption resin to adsorb impurities, and an effluent and a washing liquid are collected;

[0009] (3) purifying the effluent and the washing liquid in step (2) through an organic solvent nanofiltration membrane to obtain a decolorized retained membrane liquid; the organic solvent nanofiltration membrane comprises a first-stage organic solvent nanofiltration membrane and a second-stage organic solvent nanofiltration membrane, the first-stage organic solvent nanofiltration membrane is an organic solvent nanofiltration membrane with a molecular weight of 800 to 1500 Daltons, and the second-stage organic solvent nanofiltration membrane is an organic solvent nanofiltration membrane with a molecular weight of 100 to 250 Daltons;

[0010] (4) the decolorized retained membrane liquid is fed into an anion exchange resin, and then eluted with pure water, and the effluent and the washing liquid are collected, and then fed into a cation exchange resin, and then eluted with pure water, and the effluent and the washing liquid are collected;

[0011] (5) passing the effluent and the water washing liquid collected after passing through the cation exchange resin in step (4) through a reverse osmosis membrane to obtain a retentate;

[0012] (6) The retentate is concentrated into an extract, dissolved in an organic solvent, stirred evenly, and acid is added to adjust the pH to acidic, cooled for crystallization, and dried to obtain a quebracho alcohol product.

[0013] In step (1) of the present invention, the seabuckthorn leaves are ultrafinely ground to a size of more than 200 meshes, the mass ratio of the seabuckthorn leaves to pure water is 1:5-8, and the extraction time is 3-5 hours.

[0014] In the present invention, the term "at room temperature" refers to 15 to 25°C.

[0015] In step (1) of the present invention, the graded centrifugation is that the filtrate is first subjected to butterfly centrifugation at 1500-2500 r / min for 25-40 min, and then subjected to tubular centrifugation at 9000-11000 r / min for 25-40 min.

[0016] In step (1) of the present invention, the pressurized pressure is 1 to 3 MPa.

[0017] In step (2) of the present invention, the macroporous adsorption resin is D101, LX-T28, LX-8, ADS-7, LX-20B or LX-T81.

[0018] In step (3) of the present invention, the membrane inlet pressure of the first-stage organic solvent nanofiltration membrane is 290-870 psi, and the membrane inlet pressure of the second-stage organic solvent nanofiltration membrane is 500-850 psi.

[0019] In step (4) of the present invention, the anion exchange resin is D941, D945, D296, D330, D918 or LX-T5; the cation exchange resin is 001*16MB, SQ66, SQD-65, 001*7MB or D110.

[0020] In step (5) of the present invention, the pressure when passing through the reverse osmosis membrane is 0.8-1 MPa.

[0021] In step (6) of the present invention, the organic solvent is methanol or ethanol.

[0022] In step (6) of the present invention, the acid is citric acid or hydrochloric acid, and the acid is added to adjust the pH to 3.0-4.0.

[0023] The present invention has the following beneficial effects:

[0024] (1) The content of quebracho alcohol extracted by the extraction method of the present invention is as high as 97%, and the recovery rate is as high as 80%. The extraction method is safe, environmentally friendly, and easy to operate, and is suitable for large-scale industrial production.

[0025] (2) The present invention ultrafinely grinds seabuckthorn leaves to promote the dissolution rate of quebracho alcohol and increase the content of quebracho alcohol in the extract; in addition, the present invention extracts at room temperature without heating, which can effectively reduce production costs and improve production efficiency.

[0026] (3) The present invention adopts a combination of a macroporous adsorption resin and an organic solvent nanofiltration membrane for impurity removal and purification. The macroporous adsorption resin can effectively adsorb organic matter; the organic solvent nanofiltration membrane can effectively intercept organic matter with a certain molecular weight, thereby playing a role in separating products and impurities, while taking into account the decolorization effect. The above-mentioned combination of impurity removal and purification is simple and convenient to operate, consumes less energy, and has a large processing capacity.

[0027] (4) The present invention uses anion exchange resin and cation exchange resin to remove some soluble proteins and amino acids for further graded purification; and the use of ion exchange resin takes into account the effects of decolorization, purification and pH neutralization.

[0028] (5) The present invention uses a reverse osmosis membrane to remove organic salts and small molecules, such as monosaccharides, thereby further increasing the content of the product and facilitating the next step of crystallization. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the chromatogram of the standard product of quebracho alcohol;

[0030] Figure 2 The figure is the chromatogram of the product quebracho alcohol of the present invention. DETAILED DESCRIPTION

[0031] Example 1

[0032] (1) Take 1000g of dried seabuckthorn leaves, wherein the total content of quebracho alcohol is 3.0%, and ultrafinely grind to more than 200 meshes, add 5 times amount of pure water and stir evenly, keep stirring and extracting at room temperature, the extraction time is 4 hours, and the extraction is done twice. After extraction, pass through 200 mesh filter cloth, and then use desktop centrifuge for graded centrifugation, first through 2000r / min butterfly low-speed centrifugation for 30min, and then through 10000r / min tubular high-speed centrifugation for 30min; the filter residue is pressed dry under atmospheric pressure of 1MPa to obtain filter residue filtrate, and mixed with the filtrate after graded centrifugation to obtain extract.

[0033] (2) The extract was injected into 500 g of macroporous adsorption resin LX-T28 at a drug injection rate of 1 BV / h. After the injection, it was eluted with 2 times the column volume of pure water, and the effluent and water washing solution were collected and combined.

[0034] (3) Purifying the effluent and the washing liquid in step (2) through an organic solvent nanofiltration (OSN) membrane, wherein the organic solvent nanofiltration membrane comprises a first-stage organic solvent nanofiltration membrane and a second-stage organic solvent nanofiltration membrane, wherein the first-stage organic solvent nanofiltration membrane is an organic solvent nanofiltration membrane with a molecular weight of 1000 Daltons and a membrane inlet pressure of 290 to 870 psi; and the second-stage organic solvent nanofiltration membrane is an organic solvent nanofiltration membrane with a molecular weight of 200 Daltons and a membrane inlet pressure of 500 to 850 psi, to obtain a decolorized retained membrane liquid.

[0035] (4) The decolorized retained membrane liquid was fed with 500 g of anion exchange resin D330 at a feeding rate of 0.5 BV / h. After the feeding, it was eluted with 2 times the column volume of pure water, and the effluent and the washing liquid were collected together; then it was directly fed with 300 g of cation exchange resin 001*16MB at a feeding rate of 1 BV / h. After the feeding, it was eluted with 2 times the column volume of pure water, and the effluent and the washing liquid were collected together.

[0036] (5) The effluent and water washing liquid from the cation exchange resin in step (4) are passed through a reverse osmosis membrane at a pressure of 1 MPa. After the original liquid is passed through, pure water 4 times the amount of the original liquid is added to obtain a retentate.

[0037] (6) The retentate is concentrated to an extract with a solid content of 70% for standby use.

[0038] (7) Add 2 times the amount of analytical pure methanol as the extract, stir evenly, adjust the pH to 3.0-4.0 with 30% (w / w) citric acid, then crystallize at 4°C for 12 hours, dry the crystals, and obtain 24.88 g of quebracho alcohol product. After HPLC detection, the content of the quebracho alcohol product was 97.57%, and the recovery rate was 80.92%.

[0039] Example 2

[0040] The difference between this embodiment and embodiment 1 is that: in step (1), 8 times the amount of pure water is added and stirred evenly, the extraction time is 3 hours, and the extraction is performed twice, followed by butterfly type low-speed centrifugation at 1500 r / min for 40 min, and then tubular high-speed centrifugation at 9000 r / min for 40 min, and the filter residue is pressed dry under atmospheric pressure of 3 MPa; in step (2), the type of macroporous adsorption resin is D101; in step (3), the molecular weight of the first-stage organic solvent nanofiltration membrane is 1200 Dalton, the molecular weight of the second-stage organic solvent nanofiltration membrane is 150 Dalton; in step (4), the type of anion exchange resin is D941, and the type of cation exchange resin is SQ66; in step (5), the pressure through the reverse osmosis membrane is 0.8Mpa; in step (6), the retentate is concentrated to an extract with a solid content of 65%; in step (7), 2 times the amount of analytical pure ethanol as the extract is added, and the pH is adjusted to 3.0-4.0 with 30% (w / w) hydrochloric acid, followed by crystallization at 0°C for 12h. 25.18g of quercetin product is obtained. After HPLC detection, the content of the quercetin product is 97.12%, and the recovery rate is 81.5%.

[0041] Example 3

[0042] The difference between this embodiment and embodiment 1 is that: in step (1), the extraction time is 5 hours, and the extraction is performed twice, followed by butterfly low-speed centrifugation at 2500 r / min for 25 minutes and then tubular high-speed centrifugation at 11000 r / min for 25 minutes; in step (2), the type of macroporous adsorption resin is LX-8; in step (3), the molecular weight of the first-stage organic solvent nanofiltration membrane is 1500 Daltons, and the molecular weight of the second-stage organic solvent nanofiltration membrane is 100 Daltons; in step (4), the type of anion exchange resin is D945, and the type of cation exchange resin is SQD-65; in step (6), the retentate is concentrated to an extract with a solid content of 75%; in step (7), twice the amount of analytical pure ethanol as the extract is added, and the pH is adjusted to 3.0-4.0 with 30% (w / w) hydrochloric acid, followed by crystallization at 6°C for 12 hours. 24.57 g of quebracho alcohol product was obtained. After HPLC detection, the content of the quebracho alcohol product was 99.17% and the recovery rate was 81.22%.

[0043] Example 4

[0044] The difference between this embodiment and embodiment 1 is that in step (2), the macroporous adsorption resin is ADS-7; in step (3), the molecular weight of the first-stage organic solvent nanofiltration membrane is 800 Daltons, and the molecular weight of the second-stage organic solvent nanofiltration membrane is 250 Daltons; in step (4), the anion exchange resin is D296, and the type of the cation exchange resin is 001*7MB. 24.54 g of quebracho alcohol product was obtained. After HPLC detection, the content of the quebracho alcohol product was 98.27%, and the recovery rate was 80.38%.

[0045] The foregoing embodiments are merely illustrative and are used to explain some features of the method of the present invention. The appended claims are intended to require the widest possible range that can be imagined, and the embodiments presented herein are demonstrated by the applicant's actual test results. Therefore, the applicant's intention is that the appended claims are not limited by the selection of examples that illustrate the features of the present invention. Some numerical ranges used in the claims also include sub-ranges within them, and changes in these ranges should also be interpreted as being covered by the appended claims where possible.

Claims

1. A method for extracting quebracho alcohol from seabuckthorn leaves, characterized in that: The following steps are involved: (1) Ultrafine grinding of seabuckthorn leaves, adding pure water and stirring, extracting at room temperature, filtering the extracted seabuckthorn leaves through filter cloth, fractionating and centrifuging the filtrate, pressing and drying the filter residue to obtain an extract; (2) the extract is passed through a macroporous adsorption resin to adsorb impurities, and an effluent and a washing liquid are collected; (3) purifying the effluent and the washing liquid in step (2) through an organic solvent nanofiltration membrane to obtain a decolorized retained membrane liquid; the organic solvent nanofiltration membrane comprises a first-stage organic solvent nanofiltration membrane and a second-stage organic solvent nanofiltration membrane, the first-stage organic solvent nanofiltration membrane is an organic solvent nanofiltration membrane with a molecular weight of 800 to 1500 Daltons, and the second-stage organic solvent nanofiltration membrane is an organic solvent nanofiltration membrane with a molecular weight of 100 to 250 Daltons; (4) the decolorized retained membrane liquid is fed into an anion exchange resin, and then eluted with pure water, and the effluent and the washing liquid are collected, and then fed into a cation exchange resin, and then eluted with pure water, and the effluent and the washing liquid are collected; (5) passing the effluent and the water washing liquid collected after passing through the cation exchange resin in step (4) through a reverse osmosis membrane to obtain a retentate; (6) The retentate is concentrated into an extract, dissolved in an organic solvent, stirred evenly, and acid is added to adjust the pH to acidic, cooled for crystallization, and dried to obtain a quebracho alcohol product.

2. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (1), the seabuckthorn leaves are ultrafinely ground to a size of more than 200 meshes, the mass ratio of the seabuckthorn leaves to pure water is 1:5-8, and the extraction time is 3-5 hours.

3. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (1), the graded centrifugation is that the filtrate is first subjected to butterfly centrifugation at 1500-2500 r / min for 25-40 min, and then subjected to tubular centrifugation at 9000-11000 r / min for 25-40 min.

4. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (1), the pressurized pressure is 1 to 3 MPa.

5. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (2), the macroporous adsorption resin is D101, LX-T28, LX-8, ADS-7, LX-20B or LX-T81.

6. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (3), the membrane inlet pressure of the first-stage organic solvent nanofiltration membrane is 290 to 870 psi, and the membrane inlet pressure of the second-stage organic solvent nanofiltration membrane is 500 to 850 psi.

7. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (4), the anion exchange resin is D941, D945, D296, D330, D918 or LX-T5; the cation exchange resin is 001*16MB, SQ66, SQD-65, 001*7MB or D110.

8. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (5), the pressure when passing through the reverse osmosis membrane is 0.8 to 1 MPa.

9. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (6), the retentate is concentrated into an extract having a solid content of 65% to 75%.

10. The method for extracting quesol from seabuckthorn leaves according to claim 1, characterized in that: In the step (6), the acid is citric acid or hydrochloric acid, and the acid is added to adjust the pH to 3.0-4.0.

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