CN111544494A - HPEF extraction method of tea seed phenolic acid - Google Patents
HPEF extraction method of tea seed phenolic acid Download PDFInfo
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Abstract
The invention belongs to the field of deep processing of tea seeds, and particularly discloses an HPEF extraction method of tea seed phenolic acid, which specifically comprises the following steps: processing raw materials, extracting by HPEF and adsorbing by resin to obtain the tea seed phenolic acid product. The invention also solves the problem that the existing extraction method has low efficiency, low purity and high cost, thereby causing the reduction of DPPH free radical scavenging capacity of phenolic acid products. Compared with the traditional ethanol water solution extraction method, the DPPH free radical scavenging capacity of the tea seed phenolic acid obtained by the method is improved by 1.37-2.08 times, and the DPPH free radical scavenging capacity is obviously improved.
Description
Technical Field
The invention belongs to the technical field of deep processing of tea seeds, and particularly relates to an HPEF extraction method of tea seed phenolic acid.
Background
Tea tree (Camellia sinensis (L.) O.Ktze.) is a plant of the genus Camellia of the family Theaceae and originated from Yunyuan plateau of China. The tea leaf, fruit and other plant materials are rich in tea polyphenols. Phenolic compounds are important secondary metabolites in plants, and the molecular structure of the phenolic compounds contains a plurality of phenolic hydroxyl groups, and the phenolic compounds are generally divided into phenolic acids, flavonoids, tannins, flavanols, glycosides and the like. The pharmacopoeia of the people's republic of China shows that the phenolic acid compounds in the tea seeds have the functions of reducing blood fat, resisting coagulation, resisting oxidation, benefiting gallbladder, resisting cancer and the like, and the content of single components in the tea seeds is measured by adopting a high performance liquid chromatography in the prior report. The active phenolic acid substances in the tea seeds are polyhydroxy compounds, and the structural characteristics of the polyhydroxy compounds determine that the phenolic acid is easy to dissolve or soluble in water, alcohols, ethers, ketones, esters and the like, so the solvent extraction method mainly comprises two types of aqueous solvent extraction and organic solvent extraction. In addition, microwave-assisted extraction, ultrasonic-assisted extraction, biological enzymolysis extraction, supercritical fluid extraction, membrane technology extraction and the like are also provided (a method for extracting phenolic acid compounds from Chinese knotweed herb, patent number 201710418772.0; a green high-efficiency ultrasonic extraction method for preventing the phenolic acid in the citrus peel dregs from degrading, patent number 201710432484.0; a phenolic acid compound in lavender, a preparation method and application thereof, patent number 201710131035.2). Tea seed phenolic acids are used as structural materials and energy materials in organisms, and more importantly, are also involved in various activities of cells in life phenomena. Phenolic acid is widely applied to the fields of food, textile, cosmetics, medicine and the like, and the important significance of the phenolic acid in biology determines that the phenolic acid has wide prospects in the fields of food and medicine.
The high-voltage pulse electric field (HPEF) method is a new technology developed in recent years, the mechanism of the method is that the cell wall is instantaneously broken by utilizing the cell membrane electroporation principle, so that the cell membrane potential is disordered, and the cell wall and the cell membrane are damaged reversibly or slightly reversibly, so that cell components flow out, and the method has the advantages of short treatment time, low energy consumption, less possibility of causing target product denaturation and the like. The HPEF extraction method aiming at improving the antioxidant activity of the tea seed phenolic acid can overcome the adverse effect of the conventional ethanol water extraction on active substances; the solvent in contact with the sample can be strengthened and the cell components in the tea seeds can be destroyed by HPEF extraction method, and the needed compound is separated from the sample matrix and then enters the solvent (a method for extracting purslane polysaccharide by using high-voltage pulse electric field, the patent number is 201310569144.4; a method for extracting citrus essential oil, the patent number is 201610121908.7; a method for extracting rice bran oil by subcritical fluid assisted by high-voltage pulse electric field, the patent number is 201410289492.0; a method for extracting burdock polysaccharide from burdock roots, the patent number is 201510389114.4).
Disclosure of Invention
In order to solve the problems of low efficiency, low purity and high cost of the existing extraction method of the tea seed phenolic acid, which result in low DPPH free radical scavenging capacity of the phenolic acid product, the invention provides the HPEF extraction method of the tea seed phenolic acid, which not only improves the extraction efficiency, but also improves the product yield, purity and DPPH free radical scavenging capacity.
The technical scheme adopted by the invention is as follows:
an HPEF extraction method of tea seed phenolic acid comprises the following steps:
(1) raw material treatment: drying tea seeds, crushing and sieving to obtain tea seed powder, and dissolving the tea seed powder in an ethanol water solution to obtain a mixture;
(2) HPEF extraction: HPEF treatment is carried out on the mixture for 2-6 h under the conditions of 400-600W of power, 20-30 KV/cm of electric field intensity, 7-11 mus of pulse width and 40-68 Hz of frequency, pressure reduction filtration is carried out, and filter residues are washed to obtain a crude product of the tea seed phenolic acid;
(3) resin adsorption: diluting the crude product of the tea seed phenolic acid, adsorbing by using macroporous resin, oscillating in water bath, filtering and washing the macroporous resin obtained by adsorption, dynamically eluting by using 55-65% ethanol after the resin is dried by filtration, concentrating the eluent, and performing vacuum freeze drying to obtain the tea seed phenolic acid.
According to the invention, through a large number of tests, the phenolic acid in different raw materials is found to have differences in dissolution rate and interference components, and the fact that the phenolic acid in the tea seeds is extracted by adopting the HPEF extraction method is unexpectedly found to greatly improve the extraction efficiency, the product yield and purity and the DPPH free radical scavenging capacity, so that the method is very suitable for industrial large-scale production and application.
In the step (1), the tea seeds are dried to constant weight and the water content of the tea seeds is lower than 0.05%, and then the tea seeds are crushed and sieved by a sieve of 90-110 meshes to obtain tea seed powder.
Preferably, in the step (1), the volume concentration of the ethanol aqueous solution is 45-55%. The dosage of the ethanol water solution is 1 mg: (50-70) mL. Experiments show that under the conditions of optimal dosage and concentration of the ethanol aqueous solution, the extraction efficiency of the tea seed phenolic acid is higher.
Preferably, in step (2), the HPEF processing conditions are: the power is 450-550W, the electric field intensity is 24-26 KV/cm, the pulse width is 8-10 mus, and the frequency is 50-60 Hz. Experiments show that under the condition of the optimized HPEF parameter, the dissolved impurity components are less in the extraction process, and the dissolved effective components are higher.
And (2) carrying out reduced pressure filtration for 50-70 min under the condition of 0.01MPa, washing the obtained filter residue for 2-3 times by using an ethanol water solution with the volume concentration of 45-55%, and combining the filter residues to obtain a crude product of the tea seed phenolic acid. The dosage of the ethanol water solution is 0.001 g: (30-80) mL.
In the step (3), the crude product of the tea seed phenolic acid is diluted by adding water to obtain a solution with the concentration of 9.5-10.5 mg/mL. The conditions of the water bath oscillation are as follows: and then oscillating for 7-9 h in a constant temperature water bath at 20-30 ℃, wherein the oscillation frequency is 1000-1400 r/min.
In the step (3), the flow rate of the macroporous resin adsorption is 2.0-4.0 BV/h, and the adsorption time is 1.0-3.0 h. The dynamic elution rate is 2.0-4.0 BV/h.
Compared with the prior art, the invention has the following beneficial effects:
(1) the extraction method is suitable for industrial production, has high extraction rate, and further improves the yield and the purity of the product;
(2) compared with the traditional ethanol water solution extraction method, the DPPH free radical scavenging capacity of the tea seed phenolic acid obtained by the method is improved by 1.37-2.08 times, and the DPPH free radical scavenging capacity of the product is obviously improved;
(3) the solvent and other reagents used in the invention have the characteristics of no toxicity, no harm, small dosage and low cost, and the invention has the advantages of simple process, no environmental pollution, high efficiency, time saving, low cost and the like, and is a green and environment-friendly tea seed phenolic acid extraction technology with wide prospect.
Drawings
FIG. 1 is a diagram showing the purity, yield and DPPH free radical scavenging ability of the extracted tea seed phenolic acid according to the present invention.
Detailed Description
In order to facilitate an understanding of the invention, the invention will be described more fully and in detail below with reference to the accompanying drawings and preferred embodiments, but the scope of the invention is not limited to the specific embodiments below.
Unless otherwise defined, all terms of art used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. Unless otherwise specified, the reagents and materials used in the present invention are commercially available products or products obtained by a known method.
Example 1
(1) Raw material treatment:
weighing tea seeds, placing the tea seeds in a 55 ℃ oven to dry until the tea seeds are nearly dry, then placing the tea seeds in a 110 ℃ constant temperature and humidity dryer to dry until the tea seeds are constant in weight, and sieving the tea seeds with a 100-mesh sieve to obtain tea seed powder (a product I A) for later use;
(2) HPEF extraction:
taking a product A obtained in the step (1) according to the ratio of the mass of the tea seed powder in mg to the volume of the ethanol water solution in mL of 1:60, placing the product A in the ethanol water solution with the volume concentration of 50%, treating the product A for 4 hours by using HPEF treatment equipment (purchased from Fuguangjiu optical refining apparatus (China)) under the treatment conditions of 500W of power, 25KV/cm of electric field intensity, 9 mus of pulse width and 60Hz of frequency, filtering the product A under the condition of 0.01MPa of circulating water vacuum pump of SHZ-D (III) for 1.0 hour, washing the filter residue for 3 times by using an ethanol extraction solvent with the concentration of 50%, and combining the washed filter residues to obtain a crude product of the tea seed phenolic acid (product II A), wherein the ratio of the mass in g to the volume of the extraction solvent in mL is 0.001:50 g/mL;
(3) resin adsorption:
adding water into the product II A obtained in the step (2) to prepare a solution of 10.0mg/mL, adsorbing by Daion HP20 macroporous resin at the flow rate of 3.0BV/h and the adsorption time of 3.0h, oscillating in a TS-100B constant temperature oscillator water bath for 8h, keeping the water bath temperature at 25 ℃ and the oscillation frequency at 1200r/min to obtain a product III A;
(4) and (3) preparing a finished product:
filtering the Daion HP20 macroporous resin after adsorbing the product III A in the step (3), washing the filtered resin twice with distilled water, dynamically eluting with ethanol with the volume concentration of 60% after the resin is dried, wherein the elution rate is 2.0BV/h, concentrating the eluent to 10% of the original volume at the temperature of 60 ℃, and then carrying out vacuum freeze drying at the temperature of-20 ℃ and the pressure of 0.1MPa to obtain the tea seed phenolic acid product (product IV A).
Example 2
(1) Raw material treatment:
weighing tea seeds, placing the tea seeds in a 55 ℃ oven to dry until the tea seeds are nearly dry, then placing the tea seeds in a 110 ℃ constant temperature and humidity dryer to dry until the tea seeds are constant in weight, and sieving the tea seeds with a 100-mesh sieve to obtain tea seed powder (product A B) for later use;
(2) HPEF extraction:
according to the ratio of the mass of the tea seed powder in mg to the volume of the ethanol aqueous solution in mL of 1:60, placing the product I B obtained in the step (1) in the ethanol aqueous solution with the volume concentration of 50%, treating for 4h by using HPEF equipment (HPEF treatment equipment of Fuguangjing optical instruments (China) Co., Ltd.) under the treatment conditions of 600W of power, 20KV/cm of electric field intensity, 7 mus of pulse width and 50Hz of frequency, filtering for 1.0h under the condition of 0.01MPa of circulating water vacuum pump of SHZ-D (III), washing the filter residue for 3 times by using the ethanol extraction solvent with the concentration of 50%, and combining the washed filter residues to obtain a crude product of the tea seed phenolic acid (product II B);
(3) resin adsorption:
adding water into the product II B obtained in the step (2) to prepare a solution of 10.0mg/mL, adsorbing by Daion HP20 macroporous resin at the flow rate of 3.0BV/h and the adsorption time of 3.0h, oscillating in a TS-100B constant temperature oscillator water bath for 8h, keeping the water bath temperature at 25 ℃ and the oscillation frequency at 1200r/min to obtain a product III B;
(4) and (3) preparing a finished product:
filtering the Daion HP20 macroporous resin adsorbing the product III B in the step (3), washing the filtered resin twice with distilled water, dynamically eluting with ethanol with the volume concentration of 60% after the resin is dried, wherein the elution rate is 2.0BV/h, concentrating the eluent at 60 ℃ to 10% of the original volume, and then carrying out vacuum freeze drying at-20 ℃ and 0.1MPa to obtain the tea seed phenolic acid product (product IV B).
Example 3
(1) Raw material treatment:
weighing tea seeds, placing the tea seeds in a 55 ℃ oven to dry until the tea seeds are nearly dry, then placing the tea seeds in a 110 ℃ constant temperature and humidity dryer to dry until the tea seeds are constant in weight, and sieving the tea seeds by a 100-mesh sieve to obtain tea seed powder (a product I C) for later use;
(2) HPEF extraction:
taking a product C obtained in the step (1) according to the ratio of the mass of the tea seed powder in mg to the volume of the ethanol water solution in mL of 1:60, placing the product C in the ethanol water solution with the volume concentration of 50%, treating the product C for 4 hours by using HPEF equipment (HPEF treatment equipment of Fuguangjie, China) under the treatment conditions of 400W of power, 30KV/cm of electric field strength, 11 Mus of pulse width and 50Hz of frequency, filtering the product C for 1.0 hour under the condition of 0.01MPa of circulating water vacuum pump of SHZ-D (III), washing the filter residue for 3 times by using ethanol extraction solvent with the concentration of 50%, and combining the washed filter residues to obtain a crude product of the tea seed phenolic acid (product II C), wherein the ratio of the mass of the filter residue in g to the volume of the extraction solvent in mL is 0.001:50 g/mL;
(3) resin adsorption:
adding water into the product II C obtained in the step (2) to prepare a solution of 10.0mg/mL, adsorbing by Daion HP20 macroporous resin at the flow rate of 3.0BV/h and the adsorption time of 2.0h, oscillating in a TS-100B constant temperature oscillator water bath for 8h, keeping the water bath temperature at 25 ℃ and the oscillation frequency at 1200r/min to obtain a product III C;
(4) and (3) preparing a finished product:
filtering the Daion HP20 macroporous resin after adsorbing the product III C in the step (3), washing the filtered resin twice with distilled water, dynamically eluting with ethanol with the volume concentration of 60% after the resin is dried, wherein the elution rate is 2.0BV/h, concentrating the eluent to 10% of the original volume at the temperature of 60 ℃, and then carrying out vacuum freeze drying at the temperature of-20 ℃ and the pressure of 0.1MPa to obtain the tea seed phenolic acid product (product IV C).
Example 4
(1) Raw material treatment:
weighing tea seeds, placing the tea seeds in a 55 ℃ oven to dry until the tea seeds are nearly dry, then placing the tea seeds in a 110 ℃ constant temperature and humidity dryer to dry until the tea seeds are constant in weight, and sieving the tea seeds by a 100-mesh sieve to obtain tea seed powder (a product I D) for later use;
(2) HPEF extraction:
according to the ratio of the mass of the tea seed powder in mg to the volume of the ethanol water solution in mL of 1:60, placing the product I D obtained in the step (1) in the ethanol water solution with the volume concentration of 50%, treating for 4h by using HPEF equipment (HPEF treatment equipment of Fuguangjing optical instruments (China) Co., Ltd.) under the treatment conditions of 400W of power, 20KV/cm of electric field intensity, 7 mus of pulse width and 40Hz of frequency, filtering for 1.0h under the condition of 0.01MPa of circulating water vacuum pump of SHZ-D (III), washing the filter residue for 3 times by using the ethanol extraction solvent with the concentration of 50%, and combining the washed filter residues to obtain a crude product of the tea seed phenolic acid (product II D);
(3) resin adsorption:
adding water into the product II D obtained in the step (2) to prepare a solution of 10.0mg/mL, adsorbing by Daion HP20 macroporous resin at the flow rate of 3.0BV/h and the adsorption time of 1.0h, oscillating in a TS-100B constant temperature oscillator water bath for 8h, keeping the water bath temperature at 25 ℃ and the oscillation frequency at 1200r/min to obtain a product III D;
(4) and (3) preparing a finished product:
filtering the Daion HP20 macroporous resin after adsorbing the product III D in the step (3), washing the filtered resin twice with distilled water, dynamically eluting with 50% ethanol by volume concentration after the resin is dried, wherein the elution rate is 2.0BV/h, concentrating the eluent to 10% of the original volume at the temperature of 60 ℃, and then carrying out vacuum freeze drying at the temperature of-20 ℃ and the pressure of 0.1MPa to obtain the tea seed phenolic acid product (product IV).
Comparative example
(1) Raw material treatment:
weighing tea seeds, placing the tea seeds in a 55 ℃ oven to dry until the tea seeds are nearly dry, then placing the tea seeds in a 110 ℃ constant temperature and humidity dryer to dry until the tea seeds are constant in weight, and sieving the tea seeds by a 100-mesh sieve to obtain tea seed powder (a product I E) for later use;
(2) traditional ethanol water extraction:
according to the proportion that the mass of the tea seed powder in mg to the volume ratio of the ethanol water solution in mL is 1:60, taking the product I E obtained in the step (1), placing the product I E in a round-bottom flask, then adding the ethanol water solution with the volume concentration of 50%, leaching for 1h in a constant-temperature water bath at 80 ℃, filtering for 1.0h under the condition of 0.01MPa of an SHZ-D (III) circulating water vacuum pump, washing the filter residue for 3 times by using the ethanol extraction solvent with the concentration of 50%, wherein the ratio of the mass of the filter residue in g to the volume of the extraction solvent in mL is 0.001:50g/mL, and combining the washed filter residues to obtain a crude product of the tea seed phenolic acid (product II E);
(3) resin adsorption:
adding water into the product II E obtained in the step (2) to prepare a solution of 10.0mg/mL, adsorbing by Daion HP20 macroporous resin at the flow rate of 3.0BV/h and the adsorption time of 3.0h, oscillating in a TS-100B constant temperature oscillator water bath for 8h, keeping the water bath temperature at 25 ℃ and the oscillation frequency at 1200r/min to obtain a product III E;
(4) and (3) preparing a finished product:
filtering the Daion HP20 macroporous resin after adsorbing the product III E in the step (3), washing the filtered resin twice with distilled water, dynamically eluting with ethanol with the volume concentration of 60% after the resin is dried, wherein the elution rate is 2.0BV/h, concentrating the eluent to 10% of the original volume at the temperature of 60 ℃, and then carrying out vacuum freeze drying at the temperature of-20 ℃ and the pressure of 0.1MPa to obtain the tea seed phenolic acid product (product IV E).
And (3) performance measurement:
the following methods and conditions were used to determine the yield, purity and DPPH free radical scavenging ability of the tea seed phenolic acids prepared according to examples 1-4 above. (antioxidant Activity)
Determination of yield and purity of phenolic acid: the purity of phenolic acid in the tea seed phenolic acid products prepared in the above examples 1 to 4 was measured by Folin-phenol method under the following conditions: accurately weighing phenolic acid product with a certain mass, adding water to dissolve, fixing the volume to a certain volume to obtain sample solution, sucking 1mL of sample solution, adding 1mL of distilled water, 1mL of Folin-phenol reagent and 3mL of 10% Na2CO3And measuring the light absorption value of the product at the wavelength of 750nm by using an ultraviolet-visible spectrophotometer, substituting the light absorption value into a standard regression equation to calculate the content of the phenolic acid in the phenolic acid product, and further calculating the purity and the yield of the phenolic acid.
The yield (mg/g) of phenolic acid is the amount of phenolic acid in the phenolic acid product (mg)/the total amount of raw materials (g).
Determination of DPPH radical scavenging ability of phenolic acid: adding 1mL of DPPH solution with the concentration of 0.1mmol/L into 1mL of sample solutions with different concentrations, standing at 370 ℃ in the dark for 20min with methanol as a blank, measuring the absorbance Ai at 517nm, adding 1mL of methanol into 1mL of DPPH solution with the concentration of 0.1mmol/L, mixing, standing at 37 ℃ in the dark for 20min, and measuring the absorbance A0 at 517nm with methanol as a blank; taking 1mL of sample solutionThe solution was mixed with 1mL of methanol, left at 37 ℃ in the dark for 20min, and the absorbance Aj was measured at 517nm with methanol as a blank. The experiment was repeated three times, and DPPH radical clearance (%) was calculated after averaging { [ a0- (Ai-Aj)/a0]× 100% ability of different extracts to scavenge free radicals IC scavenged50The values represent the concentration of extract (IC) required to calculate a free radical clearance of 50%50) Push-to-push type calculation IC5050% × mass DPPH added/mass solute in the sample solution added.
As shown in FIG. 1, it can be seen from FIG. 1 that IC of the phenolic acid of tea seeds obtained by the method of the present invention is extracted in DPPH radical system5046.23-70.23 μ g/mL, and IC of the obtained tea seed phenolic acid extracted in the comparative example50It was 82.13. mu.g/mL. Therefore, compared with the traditional 70% ethanol water extraction method, the DPPH free radical scavenging capacity of the tea seed phenolic acid prepared by the method is improved by 1.37-2.08 times, and the DPPH free radical scavenging capacity of the extracted tea seed phenolic acid is obviously improved by the method.
In conclusion, the tea seed phenolic acid prepared by the method has activities of resisting oxidation, enhancing the immune function of the body, resisting tumors and the like, and can be applied to the fields of food, medicine and tobacco.
Claims (9)
1. An HPEF extraction method of tea seed phenolic acid comprises the following steps:
(1) raw material treatment: drying tea seeds, crushing and sieving to obtain tea seed powder, and dissolving the tea seed powder in an ethanol water solution to obtain a mixture;
(2) HPEF extraction: HPEF treatment is carried out on the mixture for 2-6 h under the conditions of 400-600W of power, 20-30 KV/cm of electric field intensity, 7-11 mus of pulse width and 50-68 Hz of frequency, pressure reduction filtration is carried out, and filter residues are washed to obtain a crude product of the tea seed phenolic acid;
(3) resin adsorption: diluting the crude product of the tea seed phenolic acid, adsorbing by using macroporous resin, oscillating in water bath, filtering and washing the macroporous resin obtained by adsorption, dynamically eluting by using 55-65% ethanol after the resin is dried by filtration, concentrating the eluent, and performing vacuum freeze drying to obtain the tea seed phenolic acid.
2. The HPEF extraction method of tea seed phenolic acid according to claim 1, wherein in the step (1), the tea seeds are dried to a constant weight and the moisture content is lower than 0.05%, and then the tea seeds are crushed and sieved through a sieve of 90-110 meshes to obtain tea seed powder.
3. The HPEF extraction method of tea seed phenolic acid according to claim 1, wherein in the step (1), the volume concentration of the ethanol aqueous solution is 45-55%.
4. The method for extracting HPEF of tea seed phenolic acid according to claim 1, wherein in step (1), the amount of the aqueous ethanol solution is 1 mg: (50-70) mL.
5. The HPEF extraction method of tea seed phenolic acid according to claim 1, wherein in the step (2), the tea seed phenolic acid is subjected to reduced pressure filtration for 50-70 min under the condition of 0.01MPa, the obtained filter residue is washed for 2-3 times by using 45-55% ethanol water solution, and the filter residue is combined to obtain a crude product of the tea seed phenolic acid.
6. The HPEF extraction method of tea seed phenolic acid according to claim 1, wherein in the step (3), the crude tea seed phenolic acid is diluted with water to obtain a solution with a concentration of 9.5-10.5 mg/mL.
7. The method for extracting HPEF of tea seed phenolic acid according to claim 1, wherein in step (3), the conditions of the water bath oscillation are as follows: and then oscillating for 7-9 h in a constant temperature water bath at 20-30 ℃, wherein the oscillation frequency is 1000-1400 r/min.
8. The HPEF extraction method of tea seed phenolic acid according to claim 1, wherein in the step (3), the flow rate of the macroporous resin adsorption is 2.0-4.0 BV/h, and the adsorption time is 1.0-3.0 h.
9. The HPEF extraction method of tea seed phenolic acid according to claim 1, wherein in the step (3), the dynamic elution rate is 2.0-4.0 BV/h.
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| CN106619873A (en) * | 2016-12-29 | 2017-05-10 | 华宝香精股份有限公司 | HPEF extraction method for improving antioxidant activity of pawpaw peel polyphenol |
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| CN103012544A (en) * | 2012-12-31 | 2013-04-03 | 广西师范大学 | Method for extracting saponin and polysaccharide from tea-seed pancake |
| CN103342726A (en) * | 2013-07-16 | 2013-10-09 | 青龙高科技股份有限公司 | Preparation method and application of camellia flavonoid for reducing blood glucose |
| CN106619873A (en) * | 2016-12-29 | 2017-05-10 | 华宝香精股份有限公司 | HPEF extraction method for improving antioxidant activity of pawpaw peel polyphenol |
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