CN112851729A - Method for separating trifolioside from hesperidin waste liquid - Google Patents
Method for separating trifolioside from hesperidin waste liquid Download PDFInfo
- Publication number
- CN112851729A CN112851729A CN202011626191.4A CN202011626191A CN112851729A CN 112851729 A CN112851729 A CN 112851729A CN 202011626191 A CN202011626191 A CN 202011626191A CN 112851729 A CN112851729 A CN 112851729A
- Authority
- CN
- China
- Prior art keywords
- hesperidin
- glycoside
- waste liquid
- separating
- hovenia dulcis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/06—Benzopyran radicals
- C07H17/065—Benzo[b]pyrans
- C07H17/07—Benzo[b]pyran-4-ones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a method for separating trifolioside from hesperidin waste liquid, which comprises the following steps: s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid; s2, passing the effluent liquid in the step S1 through macroporous adsorption resin, sequentially carrying out water elution, 10-50% ethanol elution and 60-90% ethanol elution, collecting 60-90% ethanol eluate, carrying out ultrafiltration, and concentrating through a nanofiltration membrane to obtain trapped fluid; s3, cooling and crystallizing the trapped liquid in the step S2 to obtain the crude trifolioside crystal; s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into the organic mixed solution, mixing, adding activated carbon, heating, refluxing, filtering, concentrating, cooling by the concentrated solution, and crystallizing to obtain the Hovenia dulcis glycoside. The method for separating the poncirin from the hesperidin waste liquid reuses the hesperidin waste liquid, extracts the poncirin from the hesperidin waste liquid, obtains economic benefit and solves the problem of sewage treatment.
Description
Technical Field
The invention relates to the field of neoponin separation, and particularly relates to a method for separating neoponin from hesperidin waste liquid.
Background
Liver cancer is one of the most common malignant tumors with high mortality rate in China and all over the world. In recent years, the search for antitumor drugs among natural drugs has gradually received much attention. Epidemiological studies have shown that the incidence of cancer can be reduced by the ingestion of fruit and vegetables rich in flavonoids. The novel ponins are flavonoid compounds mainly existing in citrus fruits, origanum and other herbs, and are thought to induce apoptosis of A549 and NCI-H460 cells through a Fas/Fas ligand apoptosis system, and a nude mouse experiment proves that the novel ponins can be used as a novel tumor chemotherapeutic agent to delay the growth of non-small cell lung cancer. Studies also find that the neoponin inhibits the proliferation of neuroblastoma and induces the apoptosis of neuroblastoma by inhibiting N-Myc and up-regulating RKIP expression, and all the studies show the potential therapeutic effect of the neoponin in the aspect of tumor resistance.
Disclosure of Invention
The invention provides a method for separating poncirin from hesperidin waste liquid, which aims to solve the technical problem that the prior art has no effective extraction method aiming at the poncirin.
The technical scheme adopted by the invention is as follows:
a method for separating Hovenia dulcis glycoside from hesperidin waste liquid comprises the following steps:
s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid;
s2, passing the effluent liquid in the step S1 through macroporous adsorption resin, sequentially carrying out water elution, 10-50% ethanol elution and 60-90% ethanol elution, collecting 60-90% ethanol eluate, carrying out ultrafiltration, and concentrating through a nanofiltration membrane to obtain trapped fluid;
s3, cooling and crystallizing the trapped liquid in the step S2 to obtain the crude trifolioside crystal;
s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into the organic mixed solution, mixing, adding activated carbon, heating, refluxing, filtering, concentrating, cooling by the concentrated solution, and crystallizing to obtain the Hovenia dulcis glycoside.
Further, the macroporous adsorption resin in the step S2 is one of HPD100 macroporous adsorption resin, D101 macroporous adsorption resin and AB-8 macroporous adsorption resin.
Further, the flow rate of the effluent liquid passing through the macroporous adsorption resin in the step S2 is 4 BV/h-6 BV/h, the volume of water elution is 2 BV-4 BV, and the flow rate of water elution is 1 BV/h-2 BV/h; the volume of the 10-50% ethanol elution is 3-4 BV, and the flow rate of the 10-50% ethanol elution is 1-2 BV/h; the volume of the 60-90% ethanol elution is 3 BV-4 BV, and the flow rate of the 60-90% ethanol elution is 1 BV/h-2 BV/h.
Further, in step S2, a ceramic membrane of 50nm to 100nm is used for ultrafiltration.
Furthermore, the nanofiltration membrane in the step S2 is a nanofiltration membrane of 100 Da-1000 Da.
Further, the trapped liquid in the step S3 is stirred and crystallized for 24 to 72 hours at the temperature of 5 to 10 ℃, and is filtered by a plate-and-frame filter to obtain the crude poncirin crystal.
Further, the mass ratio of the immature trifolioside to the organic mixed solution in the step S4 is 5-20: 1.
Further, the organic mixed solution comprises n-butyl alcohol, ethyl acetate and petroleum ether, wherein the volume ratio of the n-butyl alcohol to the ethyl acetate to the petroleum ether is (20-10) to (10-5) to 1.
Furthermore, the adding amount of the active carbon is 1 to 10 percent of the mass of the crude Hovenia dulcis glycoside crystal.
Further, in the step S4, the heating temperature is 70-80 ℃, the reflux time is 2-3 h, the mixture is concentrated to 1/2-1/3 of the volume of the filtered filtrate, and the concentrated solution is stirred and crystallized for 24-72 h at the temperature of 5-10 ℃.
The invention has the following beneficial effects:
the method for separating the neohesperidin from the hesperidin waste liquid reuses the hesperidin waste liquid, extracts the neohesperidin from the hesperidin waste liquid, obtains economic benefit, solves the problem of sewage treatment and realizes waste recycling.
The method for separating the neoponin from the hesperidin waste liquid firstly removes synephrine through strong-base cationic resin, and the synephrine belongs to impurities on the neoponin, so that the crystallization effect of the neoponin is influenced, and the synephrine belongs to alkaloid, so that the adsorption of macroporous resin on the neoponin can be reduced due to the existence of the synephrine.
The method for separating the neohesperidin from the hesperidin waste liquid adopts macroporous resin to enrich the neohesperidin and adopts a gradient elution method to remove impurities in the hesperidin mother liquid.
The method for separating the poncirin from the hesperidin waste liquid adopts an ultrafiltration method to remove impurities of the poncirin, physical treatment is carried out, energy consumption is low, no organic solvent is remained, a nanofiltration membrane is adopted to concentrate the poncirin, the physical treatment is carried out, the energy consumption is low, chemical structure change is prevented, and the recovery rate of the poncirin is improved.
The method for separating the poncirin from the hesperidin waste liquid firstly performs coarse crystallization, then adopts the organic mixed liquid to recrystallize the poncirin to obtain the poncirin with higher purity and more uniform crystal form, and adopts the organic mixed liquid to recrystallize, thus having simple operation and being easy to realize industrial production.
In addition to the objects, features and advantages described above, other objects, features and advantages of the present invention are also provided. The present invention will be described in further detail below with reference to the accompanying drawings.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the invention and, together with the description, serve to explain the invention and not to limit the invention. In the drawings:
FIG. 1 is a liquid chromatogram of a Hovenia glycoside standard of the present invention, wherein 17.461 is a Hovenia glycoside peak; and
FIG. 2 is a liquid chromatogram of Hovenia glycosides of preferred embodiment 1 of the present invention, in which 17.475 is the Hovenia glycoside peak.
Detailed Description
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to the embodiments with reference to the attached drawings.
FIG. 1 is a liquid chromatogram of a Hovenia glycoside standard of the present invention, wherein 17.461 is a Hovenia glycoside peak; FIG. 2 is a liquid chromatogram of Hovenia glycosides of preferred embodiment 1 of the present invention, in which 17.475 is the Hovenia glycoside peak.
The method for separating the trifolioside from the hesperidin waste liquid comprises the following steps of:
s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid;
s2, passing the effluent liquid in the step S1 through macroporous adsorption resin, sequentially carrying out water elution, 10-50% ethanol elution and 60-90% ethanol elution, collecting 60-90% ethanol eluate, carrying out ultrafiltration, and concentrating through a nanofiltration membrane to obtain trapped fluid;
s3, cooling and crystallizing the trapped liquid in the step S2 to obtain the crude trifolioside crystal;
s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into the organic mixed solution, mixing, adding activated carbon, heating, refluxing, filtering, concentrating, cooling by the concentrated solution, and crystallizing to obtain the Hovenia dulcis glycoside.
The method for separating the neohesperidin from the hesperidin waste liquid reuses the hesperidin waste liquid, extracts the neohesperidin from the hesperidin waste liquid, obtains economic benefit, solves the problem of sewage treatment and realizes waste recycling.
The method for separating the neoponin from the hesperidin waste liquid firstly removes synephrine through strong-base cationic resin, and the synephrine belongs to impurities on the neoponin, so that the crystallization effect of the neoponin is influenced, and the synephrine belongs to alkaloid, so that the adsorption of macroporous resin on the neoponin can be reduced due to the existence of the synephrine.
The method for separating the neohesperidin from the hesperidin waste liquid adopts macroporous resin to enrich the neohesperidin and adopts a gradient elution method to remove impurities in the hesperidin mother liquid.
The method for separating the poncirin from the hesperidin waste liquid adopts an ultrafiltration method to remove impurities of the poncirin, physical treatment is carried out, energy consumption is low, no organic solvent is remained, a nanofiltration membrane is adopted to concentrate the poncirin, the physical treatment is carried out, the energy consumption is low, chemical structure change is prevented, and the recovery rate of the poncirin is improved.
The method for separating the poncirin from the hesperidin waste liquid firstly performs coarse crystallization, then adopts the organic mixed liquid to recrystallize the poncirin to obtain the poncirin with higher purity and more uniform crystal form, and adopts the organic mixed liquid to recrystallize, thus having simple operation and being easy to realize industrial production.
In the process of preparing hesperidin, after a hesperidin raw material is extracted by alkaline water, the pH is adjusted to be neutral, a hesperidin crystal is separated out, the hesperidin crystal is obtained after filter pressing, a filtrate is a hesperidin mother liquor which can also be called as hesperidin waste liquor, and the hesperidin waste liquor is directly discharged as sewage under normal conditions.
In this embodiment, the macroporous adsorbent resin in step S2 is one of HPD100 macroporous adsorbent resin, D101 macroporous adsorbent resin, and AB-8 macroporous adsorbent resin. The new poncirus glycoside belongs to an isomer, the new poncirus glycoside is also called as amygdalin, the new poncirus glycoside belongs to flavanone compounds, HPD100 macroporous adsorption resin, D101 macroporous adsorption resin and AB-8 macroporous adsorption resin have strong selective adsorption effect on the flavanone compounds, the new poncirus glycoside is adsorbed by Van der Waals attraction, and then the new poncirus glycoside is eluted from the macroporous adsorption resin and dissolved in 60-90% ethanol eluent.
In the embodiment, the flow rate of the effluent in the step S2 passing through the macroporous adsorption resin is 4 BV/h-6 BV/h, the volume of water elution is 2 BV-4 BV, and the flow rate of water elution is 1 BV/h-2 BV/h; the volume of the 10-50% ethanol elution is 3-4 BV, and the flow rate of the 10-50% ethanol elution is 1-2 BV/h; the volume of the 60-90% ethanol elution is 3 BV-4 BV, and the flow rate of the 60-90% ethanol elution is 1 BV/h-2 BV/h. The removed hesperidin mother liquor of synephrine passes through macroporous adsorption resin, the macroporous adsorption resin adsorbs the neohesperidin, and other impurities in the hesperidin mother liquor are removed through gradient elution by the macroporous adsorption resin, such as: the citrus aurantium glycoside is characterized by comprising flavonoid impurities such as isocoryzanol, hesperidin, diosmin, hesperetin and the like, and the citrus aurantium glycoside is not easy to dissolve in low-concentration alcohol or water and is easy to dissolve in 60-90% ethanol according to a similar compatibility principle because of small polarity of the citrus aurantium glycoside, so that the citrus aurantium glycoside is eluted from the macroporous adsorption resin along with 60-90% ethanol eluent. The above 10% to 50% ethanol, 60% to 90% ethanol and the like are expressed as volume concentrations.
In this example, a 50nm to 100nm ceramic membrane was used for ultrafiltration in step S2. Passing the 60-90% ethanol eluent through a 50-100 nm ceramic membrane to remove impurities such as pigments, macromolecular substances and the like in the eluent so as to ensure the purity of the obtained poncirin.
In the embodiment, the nanofiltration membrane in the step S2 is a nanofiltration membrane of 100 Da-1000 Da. The nanofiltration membrane is used for concentrating 60% -90% ethanol eluent passing through a ceramic membrane of 50-100 nm to 1/100-1/200 of the volume of original hesperidin mother liquor so as to be convenient for precipitation of poncirin crystals. Preferably, the nanofiltration membrane is a 100 Da-500 Da nanofiltration membrane, the molecular weight of the poncirin is 594, the effect is better when the molecular weight of the poncirin is selected to be 100 Da-500 Da, impurities with the molecular weight lower than 100 Da-500 Da can be removed through the nanofiltration membrane, and the poncirin is kept in trapped fluid.
In the embodiment, the trapped liquid in the step S3 is stirred and crystallized for 24 to 72 hours at the temperature of 5 to 10 ℃, and is filtered by a plate-and-frame filter to obtain the crude poncirin crystals. The above-mentioned Hovenia dulcis glycoside adopts the cooling crystallization mode to make crystallization separation, at 5 deg.C-10 deg.C, the solubility of Hovenia glycoside is small, and the Hovenia glycoside is separated out, then it is filtered by means of plate-frame filter so as to obtain the invented crude crystal of Hovenia glycoside.
In this embodiment, the mass ratio of the Hovenia dulcis glycoside coarse crystal and the organic mixture in step S4 is 5-20: 1. And re-dissolving the crude neoponin crystal in the organic mixed solution, adding activated carbon in an auxiliary manner, and heating and refluxing to fully dissolve the crude neoponin crystal in the organic mixed solution.
Preferably, the organic mixed solution comprises n-butyl alcohol, ethyl acetate and petroleum ether, wherein the volume ratio of the n-butyl alcohol to the ethyl acetate to the petroleum ether is (20-10) to (10-5) to 1. The n-butanol, the ethyl acetate and the petroleum ether are mixed to obtain an organic mixed solution, and the organic mixed solution is mixed to ensure that the poncirin achieves the optimal crystallization effect, so that the obtained poncirin crystals have high yield and high content.
In the embodiment, the adding amount of the active carbon is 1-10% of the mass of the immature trifolioside crystal. The activated carbon can remove impurities in the organic mixed solution in which the crude neoponin crystals are dissolved and decolor the organic mixed solution, and is beneficial to the subsequent recrystallization and precipitation of the neoponin.
In the embodiment, the heating temperature in the step S4 is 70-80 ℃, the reflux time is 2-3 h, the concentrated solution is concentrated to 1/2-1/3 of the volume of the filtered filtrate, and the concentrated solution is stirred and crystallized for 24-72 h at the temperature of 5-10 ℃. The heating temperature is 70-80 ℃, and the temperature is increased to be beneficial to fully dissolving the crude neoponin crystals in the organic mixed solution, so that the crude neoponin crystals are cooled in the subsequent process to be separated out. However, the heating temperature is controlled to be 70 ℃ to 80 ℃ because the temperature is too high and part of the organic solvent is volatilized. And filtering to remove impurities and active carbon, concentrating the filtrate to 1/2-1/3 of the original volume, cooling and crystallizing, wherein the solubility of the Hovenia dulcis glycoside in the organic mixed solution is low at 5-10 ℃, the Hovenia dulcis glycoside is separated out from the organic mixed solution, and the Hovenia dulcis glycoside is obtained after refined crystal separation, and then filtering and drying, wherein the content of the Hovenia dulcis glycoside is usually 98-99%.
Removing synephrine from the hesperidin mother liquor by using strong-base cationic resin: the method comprises the following steps: s1, feeding the hesperidin mother liquor onto a strong acid cation resin, eluting with water after the sample feeding is finished, eluting with a strong base mixed liquor after the water elution is finished, collecting the eluent eluted by the strong base mixed liquor, and adjusting the pH value of the eluent to 5-7 to obtain a neutralized solution; and S2, carrying out ultrafiltration on the neutralized liquid, then concentrating the neutralized liquid through a nanofiltration membrane, cooling and crystallizing the concentrated liquid, filtering and drying to obtain synephrine. The method comprises the steps of carrying out ion exchange through strong-acid cation resin, eluting through low-concentration strong base mixed liquor, forming salt by synephrine under the strong base condition, reducing the pH value of an eluent to 5-7 to form synephrine in a free state after elution, having no strong pungent smell, having low requirements on production equipment and environment, and meeting the requirement of green production.
In this example, a C100E strongly acidic cation resin was used as the strongly acidic cation resin. The synephrine is enriched by macroporous resin, the eluent usually adopts alcohols, but the macroporous resin can not adsorb the synephrine basically and has poor synephrine adsorption effect, so that the hesperidin mother liquor can be directly passed through the macroporous adsorption resin to obtain the neoponin, and in order to prevent the residual of synephrine part and obtain the neoponin with high purity, the strong acid cation resin is firstly carried out to remove the synephrine.
In this embodiment, the alkali mixture comprises 0.05-0.5% by weight of KOH, 0.05-1% by weight of NaOH, or 0.05-0.5% by weight of Ca (OH)2At least two kinds of mixed solution of (1). By adopting strong base mixed liquor, the synephrine belongs to ephedrine in alkaloids, phenolic hydroxyl and amino simultaneously exist in a molecular structure, so the synephrine has amphiprotic property and can be combined with acid and alkali to form salt, and the synephrine and the strong base mixed liquor are combined to form salt through elution of the strong base mixed liquor, so the synephrine and the strong base mixed liquor are eluted together, the elution effect of the synephrine is improved, the pH value of the eluent is adjusted to be 5-7 by adding acid, and the acid reacts with the strong base in the eluent in the acid adding process, so that the synephrine is reduced into free synephrine from the generated salt. And a nanofiltration membrane is adopted for desalting subsequently, and a concentration effect is achieved at the same time. The strong alkali mixed liquid comprises the following components in percentage by mass0.05 to 0.5 percent of KOH, 0.05 to 1 percent of NaOH or 0.05 to 0.5 percent of Ca (OH)2The content of the strong base in the strong base mixed solution is lower because the more the content of the strong base is, the higher the concentration is, the more the impurities are eluted from the strong acid cation resin, and the lower the content of the synephrine is. The method specifically comprises the following steps: s1, loading pretreated C100E strong acid cation resin into a stainless steel cylinder, loading hesperidin mother liquor onto the C100E strong acid cation resin, wherein the loading speed of the hesperidin mother liquor is 7BV/h, after loading is finished, eluting by water, the volume of water elution is 3.3BV, the speed of water elution is 2BV/h, after water elution is finished, eluting by strong base mixed liquor, wherein the strong base mixed liquor comprises a mixed solution of 0.1% by mass of NaOH and 0.3% by mass of KOH, the volume of the strong base mixed liquor is 3.3BV, the flow rate of the strong base mixed liquor is 2BV/h, collecting eluent after eluting by the strong base mixed liquor, and adjusting the pH value of the eluent to 7 by 10% of hydrochloric acid to obtain a neutralizing solution; and S2, ultrafiltering the neutralized solution by adopting a 100nm ceramic membrane, concentrating by adopting a 200Da nanofiltration membrane, stirring the concentrated solution at 5 ℃, standing for crystallization for 12 hours, filtering by adopting a plate-and-frame filter, and drying to obtain synephrine.
Examples
The Hovenia dulcis glycoside is purchased from 14259-47-3 of Jiuzui Biotech.
The following reagents were all marketed.
Example 1
The method for separating the trifolioside from the hesperidin waste liquid comprises the following steps:
s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid;
s2, passing the 10t effluent liquid in the step S1 through 500kg of HPD100 macroporous adsorption resin, and sequentially carrying out water elution, 50% ethanol elution and 70% ethanol elution, wherein the flow rate of the effluent liquid passing through the HPD100 macroporous adsorption resin is 5BV/h, the volume of water elution is 2BV, the flow rate of water elution is 1BV/h, the volume of 50% ethanol elution is 3BV, and the flow rate of 50% ethanol elution is 1 BV/h; eluting with 70% ethanol at a volume of 3BV and a flow rate of 1BV/h, collecting 70% ethanol eluate, passing through 100nm ceramic membrane, concentrating with 1000Da nanofiltration membrane, and concentrating to 1/200 volume of hesperidin mother liquor to obtain trapped solution;
s3, stirring the trapped liquid in the step S2 at 5 ℃ for crystallization for 24 hours, and filtering through a plate-and-frame filter to obtain crude poncirin crystals;
s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into an organic mixed solution, mixing, wherein the mass ratio of the crude Hovenia dulcis glycoside crystals to the organic mixed solution is 5: 1, the organic mixed solution comprises n-butyl alcohol, ethyl acetate and petroleum ether, the volume ratio of the n-butyl alcohol to the ethyl acetate to the petroleum ether is 10: 5: 1, adding activated carbon, the adding amount of the activated carbon is 5% of the mass of the crude Hovenia dulcis glycoside crystals, heating and refluxing for 2 hours at 70 ℃, filtering, concentrating to 1/2 of the volume of filtered filtrate, stirring and crystallizing the concentrated solution for 24 hours at 10 ℃, filtering, and drying to obtain the Hovenia dulcis glycoside.
The obtained Hovenia dulcis glycoside has purity of 98.92% and 3.5 kg.
Example 2
The method for separating the trifolioside from the hesperidin waste liquid comprises the following steps:
s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid;
s2, passing the 20t effluent liquid in the step S1 through 500kg of D101 macroporous adsorption resin, and sequentially carrying out water elution, 30% ethanol elution and 80% ethanol elution, wherein the flow rate of the effluent liquid passing through the D101 macroporous adsorption resin is 4BV/h, the volume of the water elution is 4BV, the flow rate of the water elution is 2BV/h, the volume of the 30% ethanol elution is 3BV, and the flow rate of the 30% ethanol elution is 2 BV/h; eluting with 80% ethanol at a volume of 3BV and a flow rate of 1BV/h, collecting 80% ethanol eluate, passing through 50nm ceramic membrane, concentrating with 500Da nanofiltration membrane, and concentrating to 1/100 volume of hesperidin mother liquor to obtain trapped fluid;
s3, stirring the trapped liquid in the step S2 at 10 ℃ for crystallization for 24 hours, and filtering through a plate-and-frame filter to obtain crude poncirin crystals;
s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into an organic mixed solution, mixing, wherein the mass ratio of the crude Hovenia dulcis glycoside crystals to the organic mixed solution is 5: 1, the organic mixed solution comprises n-butyl alcohol, ethyl acetate and petroleum ether, the volume ratio of the n-butyl alcohol to the ethyl acetate to the petroleum ether is 15: 8: 1, adding activated carbon, the adding amount of the activated carbon is 5% of the mass of the crude Hovenia dulcis glycoside crystals, heating and refluxing for 2h at 75 ℃, filtering, concentrating to 1/2 of the volume of filtered filtrate, stirring and crystallizing for 24h at 10 ℃ of a concentrated solution, filtering, and drying to obtain the Hovenia dulcis glycoside.
The obtained Hovenia dulcis glycoside has purity of 98.3% and weight of 7.5 kg.
Example 3
The method for separating the trifolioside from the hesperidin waste liquid comprises the following steps:
s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid;
s2, passing the 20t effluent liquid in the step S1 through 500kg of AB-8 macroporous adsorption resin, and sequentially carrying out water elution, 40% ethanol elution and 70% ethanol elution, wherein the flow rate of the effluent liquid passing through the AB-8 macroporous adsorption resin is 5BV/h, the volume of water elution is 4BV, the flow rate of water elution is 2BV/h, the volume of 40% ethanol elution is 3BV, and the flow rate of 40% ethanol elution is 1 BV/h; eluting with 70% ethanol at a volume of 4BV and a flow rate of 1BV/h, collecting 70% ethanol eluate, passing through 50nm ceramic membrane, concentrating with 800Da nanofiltration membrane, and concentrating to 1/150 volume of hesperidin mother liquor to obtain trapped solution;
s3, stirring the trapped liquid in the step S2 at 10 ℃ for crystallization for 24 hours, and filtering through a plate-and-frame filter to obtain crude poncirin crystals;
s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into an organic mixed solution, mixing, wherein the mass ratio of the crude Hovenia dulcis glycoside crystals to the organic mixed solution is 5: 1, the organic mixed solution comprises n-butyl alcohol, ethyl acetate and petroleum ether, the volume ratio of the n-butyl alcohol to the ethyl acetate to the petroleum ether is 15: 8: 1, adding activated carbon, the adding amount of the activated carbon is 3% of the mass of the crude Hovenia dulcis glycoside crystals, heating and refluxing for 2 hours at 78 ℃, filtering, concentrating to 1/2 of the volume of filtered filtrate, stirring and crystallizing the concentrated solution for 30 hours at 10 ℃, filtering, and drying to obtain the Hovenia dulcis glycoside.
The obtained Hovenia dulcis glycoside has purity of 98.5% and weight of 7.1 kg.
The neoponin prepared in example 1 and a commercially available standard neoponin were subjected to chromatographic analysis.
(1) Chromatographic conditions column Diamonsil C18 column (150 mm. times.6 mm, 5 μm); the mobile phase is methanol and water which are 45: 55, and the flow rate is 1.0 mL/min; the detection wavelength is 283 nm; the column temperature is 25 ℃; the sample amount is 10 μ L, as shown in FIG. 1 and FIG. 2, FIG. 1 is a liquid chromatogram of the standard, wherein 17.461 is Hovenia glycosides peak; FIG. 2 is a liquid chromatogram of Hovenia dulcis glycoside of example 1 of the present invention, wherein 17.475 is the peak of Hovenia dulcis glycoside, which proves that Hovenia glycoside can be obtained by the present invention.
(2) Obtaining a liquid chromatogram map, analyzing the map by high performance liquid chromatography software, wherein the percentage of the peak area of the sample in all the peak areas is an area percentage report, the peak area percentage is the purity of the obtained Hovenia dulcis glycoside, and the purity of the obtained Hovenia dulcis glycoside is 98.92% by taking example 1 as an example, as shown in figure 2 and referring to the following area percentage report.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for separating trifolioside from hesperidin waste liquid is characterized by comprising the following steps:
s1, preparing a hesperidin mother solution, passing the hesperidin mother solution through strong basic cationic resin, removing synephrine, and collecting an effluent liquid;
s2, passing the effluent liquid in the step S1 through macroporous adsorption resin, sequentially carrying out water elution, 10-50% ethanol elution and 60-90% ethanol elution, collecting 60-90% ethanol eluate, carrying out ultrafiltration, and concentrating through a nanofiltration membrane to obtain trapped fluid;
s3, cooling and crystallizing the trapped liquid in the step S2 to obtain the crude trifolioside crystal;
s4, adding the crude Hovenia dulcis glycoside crystals obtained in the step S3 into the organic mixed solution, mixing, adding activated carbon, heating, refluxing, filtering, concentrating, cooling by the concentrated solution, and crystallizing to obtain the Hovenia dulcis glycoside.
2. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
the macroporous adsorption resin in the step S2 is one of HPD100 macroporous adsorption resin, D101 macroporous adsorption resin and AB-8 macroporous adsorption resin.
3. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 2,
the flow rate of the effluent liquid passing through the macroporous absorption resin in the step S2 is 4 BV/h-6 BV/h,
the volume of water elution is 2 BV-4 BV, and the flow rate of water elution is 1 BV/h-2 BV/h;
the volume of the 10-50% ethanol elution is 3-4 BV, and the flow rate of the 10-50% ethanol elution is 1-2 BV/h;
the volume of the 60-90% ethanol elution is 3 BV-4 BV, and the flow rate of the 60-90% ethanol elution is 1 BV/h-2 BV/h.
4. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
in the step S2, a ceramic membrane of 50 nm-100 nm is used for ultrafiltration.
5. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
in the step S2, the nanofiltration membrane is a nanofiltration membrane of 100 Da-1000 Da.
6. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
the trapped liquid in the step S3 is stirred and crystallized for 24 to 72 hours at the temperature of between 5 and 10 ℃, and is filtered by a plate-frame filter to obtain the crude poncirin crystal.
7. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
the mass ratio of the immature trifolioside to the organic mixed solution in the step S4 is 5-20: 1.
8. The method of claim 7, wherein the hesperidin is separated from the hesperidin-containing waste solution,
the organic mixed liquid comprises n-butyl alcohol, ethyl acetate and petroleum ether, wherein the volume ratio of the n-butyl alcohol to the ethyl acetate to the petroleum ether is (20-10) to (10-5) to 1.
9. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
the addition amount of the active carbon is 1-10% of the mass of the trifolioside coarse crystal.
10. The method for separating Hovenia dulcis glycoside from hesperidin waste liquid according to claim 1,
and in the step S4, the heating temperature is 70-80 ℃, the reflux time is 2-3 h, the mixture is concentrated to 1/2-1/3 of the volume of the filtered filtrate, and the concentrated solution is stirred and crystallized for 24-72 h at the temperature of 5-10 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011626191.4A CN112851729B (en) | 2020-12-31 | 2020-12-31 | Method for separating trifolioside from hesperidin waste liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011626191.4A CN112851729B (en) | 2020-12-31 | 2020-12-31 | Method for separating trifolioside from hesperidin waste liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112851729A true CN112851729A (en) | 2021-05-28 |
| CN112851729B CN112851729B (en) | 2022-12-02 |
Family
ID=75999453
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011626191.4A Active CN112851729B (en) | 2020-12-31 | 2020-12-31 | Method for separating trifolioside from hesperidin waste liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112851729B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101888889A (en) * | 2007-10-10 | 2010-11-17 | 朗氏科技有限公司 | Method to recover bioactive compounds |
| CN102234301A (en) * | 2011-05-06 | 2011-11-09 | 南京泽朗医药科技有限公司 | Preparation method of didymin |
| CN103536615A (en) * | 2013-11-08 | 2014-01-29 | 中国药科大学 | Preparation method of didymin and isosakuranetin, and application thereof in anti-diabetic medicine |
| CN113999272A (en) * | 2021-10-29 | 2022-02-01 | 广西医科大学 | Preparation method and application of melissoside |
-
2020
- 2020-12-31 CN CN202011626191.4A patent/CN112851729B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101888889A (en) * | 2007-10-10 | 2010-11-17 | 朗氏科技有限公司 | Method to recover bioactive compounds |
| CN102234301A (en) * | 2011-05-06 | 2011-11-09 | 南京泽朗医药科技有限公司 | Preparation method of didymin |
| CN103536615A (en) * | 2013-11-08 | 2014-01-29 | 中国药科大学 | Preparation method of didymin and isosakuranetin, and application thereof in anti-diabetic medicine |
| CN113999272A (en) * | 2021-10-29 | 2022-02-01 | 广西医科大学 | Preparation method and application of melissoside |
Non-Patent Citations (1)
| Title |
|---|
| 李俊等: "阳离子树脂交换法提取辛弗林的工艺研究", 《现代化工》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112851729B (en) | 2022-12-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110845328B (en) | Method for preparing high-purity carnosic acid from rosemary ointment byproducts | |
| CN103641718B (en) | Method for separating and purifying chlorogenic acid and 3,5-dicaffeoylquinic acid from honeysuckle flower | |
| CN113648834B (en) | Ceramic membrane and preparation method and application thereof | |
| CN102643315A (en) | Method for purifying phlorizin from apple velamen | |
| CN104529983B (en) | Corm Eleocharitis skin extracts the method for eriodictyol | |
| CN108864223A (en) | A kind of technique separating steviol glycoside parallel connection yield of chlorogenic acid from stevia rebaudian leaf | |
| CN106349324A (en) | Method for extracting and separating maslinic acid from olive leaves | |
| CN109021046B (en) | Method for simultaneously extracting quercetin and kaempferitrin from stem and leaf of momordica grosvenori | |
| CN112851729B (en) | Method for separating trifolioside from hesperidin waste liquid | |
| CN110917240B (en) | Continuous method for separating multiple effective components from cyclocarya paliurus | |
| CN112266399A (en) | High-purity separation and extraction method of epimedium extract | |
| CN107118252B (en) | Preparation method of glycyrrhizic acid | |
| CN100494194C (en) | Method of extracting reserpine from davilpepper | |
| CN112830881B (en) | Method for separating synephrine from hesperidin waste liquid | |
| CN109824658B (en) | Method for extracting, separating and purifying 3 flavonoid glycosides from clinacanthus nutans | |
| CN111777653B (en) | Preparation method of pure natural high-content neohesperidin | |
| CN108997359A (en) | A method of chlorophyll is extracted from stevioside production waste residue | |
| CN110169472B (en) | Method for extracting natural antioxidant for functional health tea | |
| CN111138400A (en) | Extraction and separation method of cannabichromene | |
| CN105061522A (en) | Preparation method of beta-1-O-galloyl-3, 6-(R)-hexahydroxy diphenoyl-D-glucose | |
| CN114426478B (en) | High-content gallic acid and preparation method thereof | |
| CN110156601B (en) | Method for preparing high-purity chlorogenic acid from stevia rebaudiana | |
| CN112079884B (en) | Preparation method of natural neohesperidin | |
| CN109232548B (en) | Method for extracting high-purity vitexin and isovitexin from sandalwood leaves | |
| CN105925008A (en) | Production method of waxberry red pigment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |