CN112225768A - Method for extracting trilobatin and phlorizin from lithocarpus litseifolius - Google Patents
Method for extracting trilobatin and phlorizin from lithocarpus litseifolius Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
-
- 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
- C07H1/08—Separation; Purification from natural products
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Abstract
The invention discloses a method for extracting trilobatin and phlorizin from lithocarpus litseifolius, which comprises the following steps: (1) processing Lithocarpus litseifolius leaves to obtain Lithocarpus litseifolius leaf powder; (2) mixing Lithocarpus litseifolius leaf powder with ethanol water solution, and extracting trilobatin and phlorizin under ultrasonic condition; (3) centrifuging the extractive solution, and collecting supernatant. The extract obtained by the extraction method of the invention contains rubusoside higher than 162.93mg/g, phlorizin higher than 32.76mg/g, trilobatin oxidation resistance higher than 53.29%, and phlorizin oxidation resistance higher than 42.90%. The extraction method effectively reduces the liquid-material ratio and the ethanol concentration, shortens the extraction time, reduces the usage amount of the organic solvent, greatly improves the extraction efficiency and simultaneously obviously improves the oxidation resistance of trilobatin and phlorizin.
Description
Technical Field
The invention relates to a method for extracting trilobatin and phlorizin from plants, in particular to a method for extracting trilobatin and phlorizin from Lithocarpus litseifolius leaves, belonging to the field of extraction of the trilobatin and phlorizin from Lithocarpus litseifolius leaves.
Background
Lithocarpus litseifolius (Hance) Chun, also known as hydrangea strigosa, is a natural plant with abundant active ingredients. The flavone is one of the main chemical components of the sweet tea, mainly comprises phlorizin and trilobatin, belongs to dihydrochalcone substances in the sweet tea flavone, and has high development value. The lithocarpus litseifolius triloba glycoside and phlorizin have the effects of reducing blood sugar, resisting oxidation, resisting cancer, resisting aging and the like, are safe and nontoxic, and have high health function application value.
So far, few research reports about the extraction method of rubus suavissimus triloba glycosides and phlorizin are reported. Among them, chinese patent publication No. CN108948104A discloses a heating reflux process for extracting trilobatin from lithocarpus litseifolius, which has the disadvantages of long production period, complex process, high cost, etc.; chinese patent publication No. CN110143988A discloses a method for extracting trilobatin from sweet tea by an adsorption method, which mainly has the problems of high cost, low purity and the like; chinese patent publication No. CN109134557A discloses a method for extracting rubusoside by soxhlet reflux, which has the main disadvantages that: the extraction times are many, the temperature is high, and the loss is easy to cause; chinese patent publication No. CN106220693A discloses a method for ultrasonic extraction of rubusoside, which has a major disadvantage of being time-consuming.
In conclusion, the existing extraction methods of rubusoside and phlorizin have various defects such as overlong extraction time, large consumption of organic solvent, high cost, low yield, low purity and the like. Therefore, the extraction process of rubusoside and phlorizin is still to be further researched and improved at present.
Disclosure of Invention
The invention mainly aims to provide a method for extracting trilobatin and phlorizin from lithocarpus litseifolius.
The above object of the present invention is achieved by the following technical solutions:
a method for extracting trilobatin and phlorizin from Lithocarpus litseifolius comprises: (1) processing Lithocarpus litseifolius leaves to obtain Lithocarpus litseifolius leaf powder; (2) mixing Lithocarpus litseifolius leaf powder with ethanol water solution, and extracting trilobatin and phlorizin under ultrasonic condition; (3) centrifuging the extractive solution, and collecting supernatant.
As a preferred embodiment, the processing method in step (1) includes: drying Lithocarpus litseifolius leaves, pulverizing, and sieving to obtain Lithocarpus litseifolius leaf powder; wherein, the drying temperature is preferably 40-60 ℃, preferably 50 ℃; the pulverizing can be grinding the dried Lithocarpus litseifolius leaves in a grinder at 10000-; preferably, the crushing is to grind the dried Lithocarpus litseifolius leaves in a grinder at 17800rpm for 1 min; the sieving can be through a 40-100 mesh sieve, preferably through an 80 mesh sieve.
In a preferred embodiment, the ratio of Lithocarpus litseifolius leaf powder to the ethanol aqueous solution in step (2) is preferably 1 (10-50), more preferably 1 (10-25), and most preferably 1 (20-22) in terms of g: ml.
In a preferred embodiment, the concentration of the ethanol aqueous solution in the present invention may be 40 to 80% (v/v), and most preferably 67 to 70%.
As a preferred embodiment, the time of the ultrasonic extraction in the step (2) can be 10 to 50min, preferably 10 to 25min, and most preferably 20 to 25 min.
As a preferred embodiment, the ultrasonic extraction power in the ultrasonic condition in the step (2) can be 270-630W, preferably 450W.
In addition, the ultrasonic extraction in step (2) can be performed at room temperature, for example, the ultrasonic extraction can be performed at a temperature of 10-35 ℃, and the ultrasonic mode is super 1s to 1 s.
According to 4 factors which are screened out when the trilobatin and the phlorizin are extracted under the ultrasonic condition and influence the content of the trilobatin and the phlorizin of the rubus suavissimus, namely the concentration of ethanol, the material-liquid ratio of tea powder to an ethanol solvent, the ultrasonic power and the ultrasonic time, single-factor tests are carried out. The content of the rubus suavissimus triloba glycosides and phlorizin is taken as an index, and a single-factor test result shows that: the concentration of ethanol is 70%, the ratio of material to liquid is 1:20, the ultrasonic power is 450W, and the extraction time is 20min, the content of rubusoside and phlorizin is high.
According to the single-factor test result, the extraction power is kept at 450W, and three factors (X) with obvious influence on the content of the rubus suavissimus triloba glycosides and phlorizin are selected1) The ratio of the extracted material to the extracted liquid (X)2) And extraction time (X)3) Taking the content of the rubus suavissimus triloba glycosides and phlorizin as an evaluation index (Y) as independent variables, and designing a three-factor and three-level analysis test by adopting a Box-Benhnken experiment. According to test results, the highest content of rubus suavissimus triloba glycosides and phlorizin in of the extract can be seen when the ethanol concentration is 69.09%, the liquid-material ratio is 17.86v/w, and the extraction time is 18.44 min; according to the investigation result of the extraction process and the production practice, the optimal extraction process during ultrasonic extraction is determined as follows: the ethanol concentration is 70%, and the feed-liquid ratio is 1:20 (g/ml), the extraction time is 20 min; 3 batches of rubus suavissimus triloba glycoside and phlorizin are prepared according to the extraction process and are subjected to verification experiments, the measurement values of the triloba glycoside and phlorizin are 162.93 +/-5.92 mu mol mg/g and 32.76 +/-2.11 mg/g, the ethanol concentration is 69.09%, the material-liquid ratio is 1: 17.86v (g/ml), when the extraction time is 18.44min, the test values of the trilobatin and phlorizin contents are 162.296mg/g and 33.9679mg/g are close, and the verification result fully proves that the obtained sweet tea extracting solution has high trilobatin and phlorizin contents and strong antioxidant activity.
Compared with the prior art, the method for extracting trilobatin and phlorizin from lithocarpus litseifolius mainly has the following remarkable effects:
(1) the method provided by the invention obviously reduces the liquid-material ratio and the ethanol concentration, reduces the use of organic solvents, and fundamentally solves a series of problems of high cost, high harm, high pollution and the like caused by the use of a large amount of organic solvents of rubus suavissimus triloba glycosides and phlorizin.
(2) Shortens the extraction time, improves the extraction efficiency, and fundamentally solves the problem of long production cycle of the general extraction technology of the rubus suavissimus triloba glycosides and phlorizin.
(3) Compared with the conventional extraction method, the method of the invention has simple operation.
(4) Compared with the conventional extraction method, the method disclosed by the invention improves the extraction amount of trilobatin and phlorizin and reduces the difficulty of later-stage purification.
(5) The rubus suavissimus glycosides and phlorizin obtained by the preparation method have high content and strong antioxidant activity, and tests prove that the rubus suavissimus glycosides content, the phlorizin content, the trilobatin antioxidant value and the phlorizin antioxidant value are respectively 63.30%, 47.28%, 39.40% and 13.27% higher than those of rubus suavissimus extracted without ultrasonic assistance (soaking method).
In a word, the extraction method of the rubus suavissimus triloba glycoside and the phlorizin remarkably reduces the liquid-material ratio and the ethanol concentration, shortens the extraction time, and reduces the usage amount of an organic solvent; the extract obtained by the extraction method of the invention contains rubusoside higher than 162.93mg/g, phlorizin higher than 32.76mg/g, trilobatin oxidation resistance higher than 53.29%, and phlorizin oxidation resistance higher than 42.90%. The obtained extract has high content of rubusoside and phlorizin, has strong antioxidant activity, improves the antioxidant value of the rubusoside and the phlorizin under the condition of greatly improving the extraction efficiency, and has practical popularization and application values.
Drawings
FIG. 1 comparison of trilobatin and phlorizin content and antioxidant value with and without ultrasonic extraction.
FIG. 2 Trifolium glycoside standard graph.
FIG. 3 phlorizin standard graph.
FIG. 4 is a graph of the effect of ethanol concentration on rubus suavissimus triloba glycoside and phlorizin content.
FIG. 5 is a graph showing the effect of feed liquid ratio on trilobatin and phlorizin content of rubus suavissimus.
FIG. 6 is a graph showing the effect of ultrasound power on rubus suavissimus triloba glycosides and phlorizin content.
FIG. 7 is a graph showing the effect of extraction time on rubus suavissimus triloba glycoside and phlorizin content.
Detailed Description
The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. It is to be understood that the described embodiments are exemplary only and are not limiting upon the scope of the invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be within the scope of the invention.
Example 1 extraction of rubus suavissimus leaf glycosides and phlorizin of the invention (bench test)
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; putting 1g of sample powder into a beaker, mixing with 20mL of ethanol aqueous solution (70%), performing ultrasonic extraction at room temperature (25 ℃) for 20min, and collecting supernatant to obtain an extracting solution, wherein the ultrasonic power is 450W.
Diluting the obtained extract by 200 times, measuring the content of rubusoside and phlorizin of the obtained diluted sample by HPLC, and measuring the antioxidant activity by a DPPH-spiking method. The determination result shows that the contents of trilobatin and phlorizin are 162.93 + -9.321 mg/g and 32.76 + -4.419 mg/g respectively; the oxidation resistance values are 53.29 +/-0.116% and 42.90 +/-0.516% respectively.
Example 2 extraction of rubus suavissimus leaf glycosides and phlorizin of the invention (bench test)
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; putting 1g of sample powder into a beaker, mixing with 20mL of ethanol aqueous solution (40%), performing ultrasonic extraction at room temperature (25 ℃) for 10min, and collecting supernatant to obtain an extracting solution, wherein the ultrasonic power is 270W.
Diluting the obtained extract by 200 times, measuring the content of rubusoside and phlorizin of the obtained diluted sample by HPLC, and measuring the antioxidant activity by a DPPH-spiking method. The determination result shows that the contents of trilobatin and phlorizin are 136.58 + -0.02 mg/g and 29.31 + -0.03 mg/g respectively.
Example 3 extraction of rubus suavissimus leaf glycosides and phlorizin of the invention (bench test)
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; putting 1g of sample powder into a beaker, mixing with 20mL of ethanol aqueous solution (70%), performing ultrasonic extraction at room temperature (25 ℃) for 30min, and collecting supernatant to obtain an extracting solution, wherein the ultrasonic power is 450W.
Diluting the obtained extract by 200 times, measuring the content of rubusoside and phlorizin of the obtained diluted sample by HPLC, and measuring the antioxidant activity by a DPPH-spiking method. The determination result shows that the contents of trilobatin and phlorizin are 142.51 + -0.15 mg/g and 30.21 + -0.03 mg/g respectively.
Example 4 extraction of rubus suavissimus leaf glycosides and phlorizin of the invention (bench test)
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; putting 1g of sample powder into a beaker, mixing with 20mL of ethanol aqueous solution (60%), performing ultrasonic extraction at room temperature (25 ℃) for 10min, and collecting supernatant to obtain an extracting solution, wherein the ultrasonic power is 270W.
Diluting the obtained extract by 200 times, measuring the content of rubusoside and phlorizin of the obtained diluted sample by HPLC, and measuring the antioxidant activity by a DPPH-spiking method. The determination result shows that the contents of trilobatin and phlorizin are 157.68 + -0.13 mg/g and 32.00 + -0.05 mg/g respectively.
EXAMPLE 5 extraction of rubus suavissimus leaf glycosides and phlorizin of the present invention (bench test)
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; putting 1g of sample powder into a beaker, mixing with 20mL of ethanol aqueous solution (70%), performing ultrasonic extraction at room temperature (25 ℃) for 10min, and collecting supernatant to obtain an extracting solution, wherein the ultrasonic power is 630W.
Diluting the obtained extract by 200 times, measuring the content of rubusoside and phlorizin of the obtained diluted sample by HPLC, and measuring the antioxidant activity by a DPPH-spiking method. The determination result shows that the contents of trilobatin and phlorizin are respectively 158.9 +/-0.46 mg/g and 30.12 +/-0.12 mg/g.
Comparative example 1
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; according to the extraction material-liquid ratio of 1:20 w/v, 20min and 70% ethanol, and extracting the rubusoside and phlorizin by non-ultrasonic assistance (soaking method, soaking for 20min), wherein the content and the antioxidant value of the rubusoside and phlorizin are 103.55 +/-0.118 mg/g, 23.40 +/-1.38 mg/g, 37.21 +/-0.43% and 32.54 +/-0.11%. Compared with the ultrasonic-assisted extraction of trilobatin and phlorizin of the rubus suavissimus, the content and the antioxidant value of the non-ultrasonic-assisted extraction (soaking method) are respectively reduced by 36.44%, 28.57%, 39.40% and 13.27%.
Comparative example 2
Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; the strigose hydrangea leaf glycosides and phlorizin are extracted by Soxhlet reflux, and the extraction process parameters are as follows: the extraction temperature is 80 ℃, the material ratio of the extracting solution is 30:1v/w, the extraction time is 20min, and the ethanol concentration is 70%. The contents and antioxidant values of trilobatin and phlorizin of the rubus suavissimus are 133.55 + -0.118 mg/g, 28.40 + -1.38 mg/g, 33.21 + -0.43% and 30.54 + -0.11%. Compared with ultrasonic-assisted extraction of trilobatin and phlorizin, the Soxhlet reflux extraction method reduces the content and antioxidant value of trilobatin and phlorizin of the sweet tea by 18.03%, 13.30%, 43.12% and 22.03% respectively.
Example 2 extraction of rubus suavissimus leaf glycosides and phlorizin of the present invention (pilot plant test)
Oven drying tender leaf (folium Hydrangeae Strigosae) of Lithocarpus litseifolii at 50 deg.C, grinding at 17800rpm for 2min, sieving with 80 mesh sieve to obtain tea powder, sealing in a dry container, and storing at 4 deg.C; mixing 200g of sample powder with 4L of ethanol aqueous solution (60%), performing ultrasonic extraction at 50 ℃ for 25min, cooling the mixture with running water, and collecting supernatant to obtain an extracting solution, wherein the ultrasonic power is 270-630W. Diluting the extract by 200 times, measuring the content of rubusoside and phlorizin in the obtained diluted sample by using an HPLC method, and measuring the antioxidant activity by using a DPPH-spiking method, wherein the content and the antioxidant value of the rubusoside and the phlorizin are 148.56 +/-0.918 mg/g, 29.26 +/-1.38 mg/g, 55.23 +/-0.11% and 47.91% respectively.
Experimental example 1 sweet tea trilobatin and phlorizin extraction process optimization experiment
1. Extraction and analysis method
(1) Oven drying tender leaf of Lithocarpus litseifolius (sweet tea) at 50 deg.C, grinding at 17800rpm for 1min, sieving with 80 mesh sieve to obtain tea powder, sealing in dry container, and storing at 4 deg.C.
(2) Putting 1g of the tea powder obtained in the step (1) into a beaker, and mixing the tea powder with ethanol aqueous solutions with different concentrations and volumes. Samples of sweet tea were extracted at room temperature (25 ℃) using an energy accumulating probe sonicator (8 mm diameter sonicator was centered in a beaker to ensure that the bottom of the head was 1.0-1.5cm from the liquid level at each treatment). The ultrasound extraction was performed at ultrasound power levels (270W, 360W, 450W, 540W, and 630W), ultrasound times (10min, 20min, 30min, 40min, and 50 min). Under certain power and time combination, the treatment is carried out by adopting a gap mode of 1s running and 1s stopping. Each experimental treatment was repeated three times. The supernatant was collected for subsequent experiments.
(3) Chromatographic conditions are as follows: an Agilent ZORBAX Eclipse XDB-C18 (4.6X 250mm, mu.5 m) chromatographic column, wherein a 0.1% formic acid water solution (A) -acetonitrile (B) is used as a mobile phase, and the proportion of the mobile phase is 0-10 min: 5% -25% of B; 10-15 min: 25% -30% of B; 15-20 min: 30% -95% of B; 20-25 min: 95% -95% of B; 25-30 min: 95% -5% of B; 30-35 min: 95% -5% of B. The flow rate is 1mL/min, the column temperature is 30 ℃, and the sample injection amount is 20 mu L; HPLC detection was carried out at a wavelength of 280 nm.
(4) Preparing standard series solutions of trilobatin and phlorizin: accurately weighing 0.002g of trilobatin and phlorizin, respectively dissolving in 1mL of methanol (50%) to obtain standard stock solutions of trilobatin and phlorizin with the concentration of 200 mug/mL, respectively transferring different volumes of the standard stock solutions of trilobatin and phlorizin to a 2mL centrifuge tube, complementing all volumes to scale by using methanol (50%), obtaining standard series solutions of trilobatin and phlorizin with different concentrations, and storing at room temperature for later use.
(5) Drawing a trilobatin and phlorizin standard curve: respectively taking 2mL of trilobatin and phlorizin standard substances with different concentrations, filtering the standard substances with a 0.22 mu m filter membrane, and respectively measuring peak areas under the wavelength of 280nm by HPLC; meanwhile, 2mL of double distilled water is used for replacing the trilobatin and phlorizin standard solution, and the blank experiment is carried out according to the method. Finally, a trilobatin and phlorizin concentration standard curve (shown in figure 2) is prepared, and a linear regression equation is obtained, which shows that the trilobatin and phlorizin standard products show good linear relation in the range of 10-120 mu g/mL.
(6) Measuring the content of trilobatin and phlorizin in the sweet tea extracting solution: respectively taking 2mL of diluted sample, filtering the diluted sample through a 0.22 mu m filter membrane, immediately carrying out HPLC, and measuring the peak area under the wavelength of 280 nm; meanwhile, replacing trilobatin and phlorizin standard solution with 0.5mL double distilled water, and carrying out a blank experiment according to the method; ) Calculating the content of trilobatin and phlorizin in the sweet tea extract according to the standard curve of the concentration of the trilobatin and the phlorizin, wherein the formula is as follows:
the trilobatin content (mg/g dry tea powder) is 10-3(A-1.3685) N V/42.331/M, wherein A is a sample peak area (mAU), N is a dilution multiple, V is a sweet tea extract volume (mL), M is a sweet tea powder dry weight (g), and the trilobatin content is expressed as mg/g DW;
phlorizin content (mg/g dry tea powder) 10-3(A +20.171) 200V/35.493/M, wherein A is a sample peak area (mAU), N is a dilution multiple, V is a sweet tea extract volume (mL), M is a sweet tea powder dry weight (g), and phlorizin content is expressed as mg/g DW;
2. optimization of extraction process of rubus suavissimus triloba glycosides and phlorizin
(1) Screening 4 factors influencing the content of the rubus suavissimus triloba glycosides and phlorizin according to the earlier research results, wherein the factors comprise ethanol concentration, the material-liquid ratio of tea powder to an ethanol solvent, ultrasonic power and time, and carrying out single-factor test. The content of trilobatin and phlorizin of the strigose hydrangea is taken as an index, and each treatment is repeated for three times.
The single factor test result shows that: the ethanol concentration is 70% (as shown in FIG. 4), the material-liquid ratio is 1:20 (as shown in FIG. 5), the ultrasonic power is 450W (as shown in FIG. 6), and the extraction time is 20min (as shown in FIG. 7), the content of rubusfolius trilobate glycoside and phlorizin is high.
(2) According to the single-factor test result, the extraction power is kept at 450W, and three factors (X) with obvious influence on the content of the rubus suavissimus triloba glycosides and phlorizin are selected1) The ratio of the extracted material to the extracted liquid (X)2) And extraction time (X)3) As independent variables, the content of rubusoside and phlorizin is taken as an evaluation index (Y), a Box-Benhnken experiment is adopted to design a three-factor and three-level analysis test, the design factors and the level of the test are shown in table 1, and the design scheme and the result of the test are shown in table 2.
TABLE 1 test design factors and levels
TABLE 2 Experimental design and results
From the test results of table 2, it can be seen that: when the ethanol concentration is 67.53%, the liquid-material ratio is 21.96v/w, and the extraction time is 20.92min, the extract has the highest content of rubusoside and phlorizin.
(3) DPPH working solution: 0.01g of DPPH was weighed out and completely dissolved in 20mL of methanol to obtain a 0.05mg/mL solution of DPPH in methanol.
(4) Measuring the oxidation resistance values of trilobatin and phlorizin by a DPPH-spiking method: and adding 1mL of diluted extract under the optimal extraction process into 0.5mL of DPPH working solution, uniformly mixing, reacting for 6min in a dark place, performing HPLC, and measuring the peak area at the wavelength of 280 nm. The antioxidant value is expressed as the rate of decrease in the peak area of the chromatogram.
Antioxidant value (%) - (A1-A2)/A1 x 100, wherein A1 is the peak area (mAU) of the extract, and A2 is the peak area (mAU) of the reaction solution of the extract and the DPPH working solution
3. Extraction Process verification experiment
According to the investigation result of the extraction process and in combination with the actual production, the optimal extraction process is determined to be that the ethanol concentration is 70%, the material-liquid ratio is 1:20 (g/ml), the extraction time is 20 min; 3 batches of rubus suavissimus triloba glycoside and phlorizin are prepared according to the extraction process and are subjected to verification experiments, the measurement values of the triloba glycoside and phlorizin are 162.93 +/-5.92 mu mol mg/g and 32.76 +/-2.11 mg/g, the ethanol concentration is 69.09%, the material-liquid ratio is 1: 17.86v (g/ml), the test values of the trilobatin and phlorizin contents are approximate to 162.296mg/g and 33.9679mg/g when the extraction time is 18.44 min. The verification result fully proves that the obtained sweet tea extracting solution has high trilobatin and phlorizin content and strong antioxidant activity.
Claims (10)
1. A method for extracting trilobatin and phlorizin from Lithocarpus litseifolius comprises: (1) processing Lithocarpus litseifolius leaves to obtain Lithocarpus litseifolius leaf powder; (2) mixing Lithocarpus litseifolius leaf powder with ethanol water solution, and extracting under ultrasonic condition; (3) centrifuging the extractive solution, and collecting supernatant.
2. The method of claim 1, wherein the processing method in step (1) comprises: oven drying Lithocarpus litseifolius leaf, pulverizing, and sieving to obtain Lithocarpus litseifolius leaf powder.
3. The method of claim 2, wherein the temperature of said drying is 40-60 ℃; the crushing is to grind the dried Lithocarpus litseifolius leaves in a grinder at 10000-; the sieving is to sieve through a 40-100 mesh sieve.
4. The method of claim 3, wherein the temperature of said drying is 50 ℃; the crushing is to grind the dried Lithocarpus litseifolius leaves in a grinder at the rotating speed of 17800rpm for 1 min; the sieving is 80-mesh sieving.
5. The method according to claim 1, wherein the ratio of Lithocarpus litseifolius leaf powder to the ethanol aqueous solution in the step (2) is 1 (10-50), preferably 1 (10-25), most preferably 1 (20-22) in terms of g: ml.
6. The method according to claim 1 or 5, wherein the aqueous ethanol solution is an aqueous ethanol solution with a concentration of 40-80%, preferably an aqueous ethanol solution with a concentration of 67-70%.
7. The method according to claim 1, wherein the time of the ultrasonic extraction in step (2) is 10-50min, preferably 10-25min, most preferably 20-25 min.
8. The method according to claim 1, wherein the ultrasonic extraction power in the ultrasonic conditions in step (2) is 270-.
9. The method according to claim 1, wherein the ultrasonic extraction in step (2) is performed at room temperature, which is a temperature of 10-35 ℃.
10. The method of any of claims 1, 8 or 9, wherein the ultrasound mode is super 1s to 1 s.
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