CN104435363A - Method for extracting polyphenol from tea seeds by adopting aqueous enzymatic process-ethanol reflux method - Google Patents
Method for extracting polyphenol from tea seeds by adopting aqueous enzymatic process-ethanol reflux method Download PDFInfo
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Abstract
The invention discloses a method for extracting polyphenol from tea seeds by adopting an aqueous enzymatic process-ethanol reflux method. The method comprises the following steps: (1) preparing tea seed powder: peeling the tea seeds, crushing, sieving and drying until the weight is constant; (2) weighing the tea seed powder, adding some water according to a material-liquid ratio of 1:(4-6), heating for deactivating an enzyme, cooling to room temperature, adding an enzyme solution and the rest water, stirring uniformly, and performing enzymolysis for 1.0-2.0 hours; (3) performing ethanol reflux extraction: adding ethanol into the tea seed powder mixed solution after the enzymolysis to ensure that the percent by volume of ethanol in the mixed solution is 40% to 60% and the material-liquid ratio is 1:(20-30), refluxing and extracting for 25-40 minutes at the temperature of 70 to 90 DEG C, cooling to room temperature, centrifuging, and taking out the centrifuged liquid supernatant for later use; washing filter residues a plurality of times by using 60% ethanol, filtering, combining filtrate with the centrifuged liquid supernatant, rotationally evaporating and drying to obtain a target product. By combining an enzyme method with an ethanol reflux extraction technology, the method disclosed by the invention can achieve a tea seed polyphenol yield which is far higher than that achieved by a single solvent extraction method, so the method has an obvious advantage in extraction of the tea seed polyphenol.
Description
Technical field
The invention belongs to the extraction field of tea polyphenols, be specifically related to the method that a kind of aqueous enzymatic method-ethanol refluxing process extracts tea-seed polyphenol.
Background technology
China is the township of Folium Camelliae sinensis, tea and tea-seed resource very abundant, but tea-seed only has small part for breeding, among most of tea-seed resource is then in for a long time and wastes.For a long time people to the research of Folium Camelliae sinensis far more than tea-seed, and not enough to the exploitation of tea-seed.Mainly concentrate in the extraction refine of tea-seed oil and the extraction of tea saponin to the research of tea-seed both at home and abroad, and it is less to the research and development of tea-seed polyphenol, tea-seed polyphenol has the effect such as antibacterial, anticancer, antiviral, defying age, blood pressure lowering blood fat reducing, apply all to some extent in every field, as agricultural, medicine, daily-use chemical industry and food industry are all used widely.Therefore, exploitation tea-seed polyphenol, avoids the wasting of resources to have very important meaning and wide prospect.
In current aqueous enzymatic extraction tea-seed, effective ingredient is a kind of method of comparatively environmental protection, but the extraction ratio of aqueous enzymatic method is also not too high, cause the wasting of resources to a certain extent, due to the extraction and purification process also imperfection of tea-seed polyphenol, therefore find the simple extraction of extracting method efficiently tea-seed polyphenol significant.
Summary of the invention
The object of this invention is to provide the method that a kind of aqueous enzymatic method-ethanol refluxing process extracts tea-seed polyphenol, aqueous enzymatic method and alcohol reflux technology are combined, significantly improves the extraction ratio of polyphenol in tea-seed.
The present invention adopts following technical scheme:
Aqueous enzymatic method-ethanol refluxing process extracts a method for tea-seed polyphenol, comprises the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) taking tea-seed powder, is that 1:4-6 first adds part water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, and heating enzyme denaturing, is then cooled to room temperature, adds enzyme liquid and residue water, stirs, regulate temperature 60 C, stir enzymolysis 1.0-2.0h;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 40%-60%, solid-liquid ratio is 1:20-30, reflux, extract, 80-100min under 70-90 DEG C of condition, after being cooled to room temperature, centrifugal 5-15min under 3500-4500r/min, takes out for subsequent use by the supernatant after centrifugal; Filtering residue with 60% ethanol purge for several times, then filter, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.
Preferably, enzyme liquid is that cellulase is dissolved in part distilled water and makes in described step (2), and the consumption of cellulase is 1-4mg/g tea-seed powder; Carry out fast under cryogenic during described enzyme liquid preparation.
In described step (2), the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor is 1:5, and enzymolysis time is 1.5h.
In described step (3) mixed liquor, the percent by volume of ethanol is 49%, and solid-liquid ratio is 1:24, and Extracting temperature is 80 DEG C.
More preferably, the consumption of described step (2) cellulase is 2mg/g tea-seed powder.
Further, a kind of aqueous enzymatic method-ethanol refluxing process extracts the method for tea-seed polyphenol, comprises the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) aqueous enzymatic extraction: take tea-seed powder, be that 1:5 first adds part water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, 100 DEG C are heated 5 minutes to eliminate self enzymatic activity of tea-seed powder, then room temperature is cooled to, add the enzyme liquid and residue water that have configured, stir, at 60 DEG C, enzymolysis 1.5h under stirring condition; Described enzyme liquid is that cellulase is dissolved in part distilled water and makes fast under cryogenic, and the consumption of cellulase is 2mg/g tea-seed powder;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 40%-60%, solid-liquid ratio is 1:24, reflux, extract, 90min under 80 DEG C of conditions, after being cooled to room temperature, centrifugal 10min under 3500r/min, takes out for subsequent use by the supernatant after centrifugal; Filtering residue 60% ethanol purge 2-3 time, then filters, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.
The beneficial effect that the present invention reaches: by aqueous enzymatic method coupling alcohol reflux technology, the yield of tea-seed polyphenol, far above the result obtained according to National Standard Method tea leaf polyphenols Determination of Gross, illustrates that adopting the method to extract tea-seed polyphenol has clear superiority.
Effect of the present invention is illustrated below by way of following experiments:
Test example 1 extracting method screens
In tea-seed polyphenol leaching process, the present inventor designs extraction process and selects, with extraction time, tea-seed polyphenol content for inspection target, take into full account that aqua method, aqueous enzymatic method, aqueous enzymatic method-ethanol reflux extraction three kinds of methods are on the impact of extraction time and polyphenol yield, have carried out a series of Selection experiment.
Sample A: prepare according to the method for " sample preparation " in GB GB/T8313-2008 " detection method of Tea Polyphenols in Tea and catechin content ";
Sample B: prepare according to the embodiment of the present invention 1 method;
Sample C: according to the embodiment of the present invention 1 method, removal step 3) preparation, namely only adopt aqueous enzymatic extraction, wherein enzymolysis time is 1.5h;
In said method, tea-seed powder raw material is 2g, and the investigation of the object product obtained the results are shown in Table 1.
Table 1 process choice experimental result
| Index | Sample A | Sample B | Sample C |
| Extraction time (h) | 1 | 2.0 | 1.5 |
| Polyphenol content (mg/g) | 6.35 | 9.42 | 5.53 |
Upper table can be found out, aqueous enzymatic method of the present invention coupling alcohol reflux technology is adopted to extract tea-seed polyphenol, tea-seed polyphenol content is much higher than sample A, namely the inventive method is significantly better than the simple extraction methods of trial-manufacture of sample in existing National Standard Method, and always consuming time compared with additive method difference little; Adopt separately aqueous enzymatic extraction tea-seed polyphenol in addition, content is starkly lower than the present invention.The present invention is described due to the alcohol reflux technology that has been coupled on the basis of aqueous enzymatic method, improve polyphenol content, is desirable extracting method.
Further research is done to each technological parameter of extraction below.
Test 2: each parameter of aqueous enzymatic extraction is on the impact of extraction ratio
2.1 enzyme additions are on the impact of polyphenol extracted amount
Accurately take the tea-seed powder that 5 parts are dried to constant weight, each 2g, adds in beaker respectively.Add the cellulase of different quality, enzyme dosage is made to be respectively 1mg/g, 2mg/g, 3mg/g, 4mg/g, 5mg/g, maintenance solid-liquid ratio is 1:5, enzymolysis 1.5h, enzymolysis under the condition that hydrolysis temperature is 60 DEG C, then adding 60% ethanol makes liquid ratio be 1:25, reflux, extract, half an hour in 80 DEG C of water-baths, centrifugal after being down to room temperature, residue 60% ethanol purge 2-3 time, filter after and supernatant merge, be settled to 100ml.Get certain volume solution and carry out polyphenol content mensuration.The results are shown in Table 2.
Table 2 enzyme addition is on the impact of polyphenol yield
| Enzyme addition | 1mg/g | 2mg/g | 3mg/g | 4mg/g | 5mg/g |
| Polyphenol yield (mg/g) | 8.09 | 8.76 | 8.43 | 7.68 | 7.22 |
As can be known from the above table, within the specific limits, tea-seed total polyphenols yield increases gradually along with the increase of enzyme amount, and when enzyme dosage reaches 2mg/g, namely yield reaches maximum, and continue afterwards to strengthen enzyme addition, tea-seed polyphenol yield declines on the contrary.Therefore select enzyme concentration scope to be 1-3mg/g, be preferably 2mg/g.
2.2 enzymolysis times are on the impact of total polyphenols extracted amount
Accurately take the tea-seed powder that 5 parts are dried to constant weight, each 2g, adds in beaker respectively.Keep enzyme addition to be 2mg/g, solid-liquid ratio is 1:5, and hydrolysis temperature is 60 DEG C, respectively enzymolysis 0.5h, 1h, 1.5h, 2h, 2.5h.Then adding 60% ethanol makes liquid ratio be 1:25, reflux, extract, half an hour in 80 DEG C of water-baths, centrifugal after being down to room temperature, residue 60% ethanol purge 2-3 time, after filtering and supernatant merge, be settled to 100ml.Get certain volume solution and carry out polyphenol content mensuration.The results are shown in Table 3.
Table 3 enzymolysis time is on the impact of polyphenol extraction ratio
| Enzymolysis time | 0.5h | 1h | 1.5h | 2h | 2.5h |
| Polyphenol yield (mg/g) | 7.92 | 8.31 | 8.63 | 8.04 | 7.15 |
As can be known from the above table, in other condition one timings, along with the increase polyphenol extraction ratio of enzymolysis time raises in 0.5-1.5h, when enzymolysis time is 1.5h, polyphenol extraction ratio is the highest, with the increase of enzymolysis time, tea-seed powder total polyphenols yield declines on the contrary, therefore selection time scope is 1-2 hour, preferably 1 hour.
2.3 solid-liquid ratios are on the impact of total polyphenols extracted amount
Accurately take the tea-seed powder that 5 parts are dried to constant weight, each 2g, adds in beaker respectively.Keep hydrolysis temperature to be 60 DEG C, enzymolysis half an hour, enzyme addition is 2mg/g, makes solid-liquid ratio (g:ml) be respectively 1:3,1:4,1:5,1:6,1:7.Then adding 60% ethanol makes liquid ratio be 1:25, reflux, extract, half an hour in 80 DEG C of water-baths, centrifugal after being down to room temperature, residue 60% ethanol purge 2-3 time, after filtering and supernatant merge, be settled to 100ml.Get certain volume solution and carry out polyphenol content mensuration.The results are shown in Table 4.
Table 4 liquid ratio is on the impact of tea-seed polyphenol extraction ratio
| Liquid ratio (g:ml) | 1:3 | 1:4 | 1:5 | 1:6 | 1:7 |
| Polyphenol yield (mg/g) | 7.85 | 8.24 | 8.73 | 8.06 | 7.30 |
As can be known from the above table, during solid-liquid ratio is 1:3 to 1:5, along with the increase tea-seed polyphenol extraction ratio of solid-liquid ratio also increases thereupon, afterwards, along with the increase of solid-liquid ratio, tea-seed total polyphenols yield declines on the contrary.Therefore select solid-liquid ratio to be 1:4-6, preferred solid-liquid ratio is 1:5.
2.4 reaction conditions
Above-mentioned experimental result draws, in enzyme digestion reaction, select enzyme concentration to be 1-3mg/g, solid-liquid ratio is 1:4-6, and the response time is 1-2h, and the Determination of Polyphenols of tea-seed is desirable. optimal conditions is: enzyme concentration 2mg/g, solid-liquid ratio 1:5, and the response time is 1.5h.
Test 3: ethanol reflux extraction is on the impact of polyphenol extraction ratio
3.1 Extracting temperature are on the impact of tea-seed polyphenol yield
Tea-seed polyphenol is the very easily oxidized compound of a class, the enzymatic oxidation of the polyphenol oxidase mainly in tea-seed, and in an experiment, has the high temperature enzyme denaturing of 5min, and in leaching process, the oxidation of tea-seed polyphenol substantially eliminates the effect of enzymatic oxidation.The non-enzymatic oxidation of tea-seed polyphenol is mainly by the impact of the factor such as temperature, time, the too high meeting of temperature makes tea-seed polyphenol oxidase, but need again to extend extraction time in extraction at cryogenic conditions, and long extraction time also can cause tea-seed polyphenol oxidase, so, should consider that the time is unsuitable long, also can not extract at an excessive temperature.This experiment have employed different temperatures and carries out extraction experiment, and method is as follows:
Accurately take the tea-seed powder that 5 parts are dried to constant weight, each 2g, after enzymolysis, be 1:20 according to liquid ratio, volume fraction of ethanol 60%, extraction time is 90min, reflux temperature is cooled to room temperature after being respectively 50 DEG C, 60 DEG C, 70 DEG C, 80 DEG C, 90 DEG C reflux, extract, proceeds to centrifuge at the centrifugal 10min of rotating speed of 3500 revs/min, supernatant is transferred to 100ml volumetric flask, residue 60% ethanol purge, transfers to 100ml volumetric flask again by filtrate, is settled to 100ml with 60% ethanol, shake up, cross 0.45 μm of film.Get certain volume solution and carry out polyphenol content mensuration.The results are shown in Table 5.
Table 5 Extracting temperature is on the impact of tea-seed polyphenol yield
| Extracting temperature | 50℃ | 60℃ | 70℃ | 80℃ | 90℃ |
| Polyphenol yield (mg/g) | 7.37 | 7.56 | 8.24 | 8.83 | 8.26 |
As seen from the above table, in alcohol reflux process, the yield of tea-seed polyphenol increases with the rising of temperature, when Extracting temperature reaches 80 DEG C, tea-seed polyphenol yield is 8.83mg/g, and along with Extracting temperature raises further, the yield of tea-seed polyphenol decreases again, be because when temperature is higher, the degree of oxidation of tea-seed polyphenol is strengthened.Therefore, selective extraction temperature be 70-90 DEG C ideal, wherein when Extracting temperature is 80 DEG C, tea-seed polyphenol yield is the highest.
3.2 concentration of alcohol are on the impact of tea-seed polyphenol yield
At present, for tea-seed polyphenol extract solvent generally have methanol, water, ethanol, acetone, etc., because tea-seed polyphenol is applied to food industry mostly, the solvent of use can not have carcinogenic and toxic and side effects.The ethanol water that this experiment have employed variable concentrations carries out extraction experiment, and method is as follows:
Accurately take the tea-seed powder that 5 parts are dried to constant weight, each 2g, after enzymolysis, be 1:20 according to liquid ratio, extraction time is 90min, reflux temperature is 70 DEG C, and volume fraction of ethanol 40%, 50%, 60%, 70%, 80%, after reflux, extract, be cooled to room temperature, proceed to centrifuge at the centrifugal 10min of rotating speed of 3500 revs/min, supernatant is transferred to 100ml volumetric flask, residue cleans by different concentration ethanol respectively, filtrate is transferred to 100ml volumetric flask again, be settled to 100ml by different concentration ethanol respectively, shake up, cross 0.45 μm of film.Get certain volume solution and carry out polyphenol content mensuration.The results are shown in Table 6.
Table 6 concentration of alcohol is on the impact of tea-seed polyphenol yield
| Concentration of alcohol | 40% | 50% | 60% | 70% | 80% |
| Polyphenol yield (mg/g) | 8.4 | 8.67 | 8.13 | 6.69 | 5.82 |
Upper table is known, in alcohol reflux process, concentration of alcohol in the scope of 40%-50%, along with the yield of the increase tea-seed polyphenol of concentration of alcohol increases, extracting solution color burn.When concentration of alcohol reaches 50%, tea-seed polyphenol yield is 8.67mg/g.Along with concentration of alcohol continues to increase, tea-seed polyphenol yield declines to some extent.Therefore, select concentration of alcohol scope to be 40-60%, when wherein concentration is 50%, tea-seed polyphenol yield is the highest.
3.3 solid-liquid ratios are on the impact of tea-seed polyphenol yield
Accurately take the tea-seed powder that 5 parts are dried to constant weight, each 2g, after enzymolysis, according to the liquid ratio of 1:10,1:15,1:20,1:25,1:30, extraction time is 90min, reflux temperature is 70 DEG C, and volume fraction of ethanol 60%, after reflux, extract, be cooled to room temperature, proceed to centrifuge at the centrifugal 10min of rotating speed of 3500 revs/min, supernatant is transferred to 100ml volumetric flask, residue 60% ethanol purge, filtrate is transferred to 100ml volumetric flask again, be settled to 100ml with 60% ethanol, shake up, cross 0.45 μm of film.Get certain volume solution and carry out polyphenol content mensuration.The results are shown in Table 7.
Table 7 solid-liquid ratio is on the impact of tea-seed polyphenol yield
| Solid-liquid ratio | 1:10 | 1:15 | 1:20 | 1:25 | 1:30 |
| Polyphenol yield (mg/g) | 6.5 | 6.89 | 8.06 | 8.42 | 6.96 |
Upper table is known, is 60% at concentration of alcohol, and Extracting temperature is under the condition of 70 DEG C, increase tea-seed polyphenol yield with solid-liquid ratio is in rising trend, and when solid-liquid ratio is 1: 25, tea-seed polyphenol yield is 8.42mg/g, but after solid-liquid ratio is more than 1: 25, tea-seed polyphenol yield declines to some extent.Therefore, select solid-liquid ratio scope to be that 1:20-30 is ideal, wherein 1: 25 time, tea-seed polyphenol yield is the highest.
Test 4: demonstration test
Adopt the optimal processing parameter of above-mentioned optimization, extract three times to tea-seed powder, its confirmatory experiment result is as shown in table 6.
Table 6 confirmatory experiment result
| Experiment number | 1 | 2 | 3 | Average content mg/g |
| Determination of Polyphenols mg/g | 9.49 | 9.51 | 9．55 | 9.52 |
Detailed description of the invention:
Embodiment 1
Aqueous enzymatic method-ethanol refluxing process extracts the method for tea-seed polyphenol, comprises the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) aqueous enzymatic extraction: take tea-seed powder 10g, be that 1:5 first adds 30ml water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, 100 DEG C are heated 5 minutes to eliminate self enzymatic activity of tea-seed powder, then room temperature is cooled to, add the enzyme liquid and residue water that have configured, stir, at 60 DEG C, enzymolysis 1.5h under stirring condition; Described enzyme liquid is that cellulase is dissolved in part distilled water and makes fast under cryogenic, and the consumption of cellulase is 2mg/g tea-seed powder;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 49%, solid-liquid ratio is 1:24, reflux, extract, 90min under 80 DEG C of conditions, to be cooled to after room temperature centrifugal 10min under 3500r/min, the supernatant after centrifugal is taken out for subsequent use; Filtering residue 60% ethanol purge 2-3 time, then filters, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.After testing, polyphenol content is 9.52mg/g.
Embodiment 2
Aqueous enzymatic method-ethanol refluxing process extracts the method for tea-seed polyphenol, comprises the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) aqueous enzymatic extraction: take tea-seed powder 10g, be that 1:4 first adds 20ml water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, 100 DEG C are heated 5 minutes to eliminate self enzymatic activity of tea-seed powder, then room temperature is cooled to, add the enzyme liquid and residue water that have configured, stir, at 60 DEG C, enzymolysis 1h under stirring condition; Described enzyme liquid is that cellulase is dissolved in part distilled water and makes fast under cryogenic, and the consumption of cellulase is 1mg/g tea-seed powder;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 40%, solid-liquid ratio is 1:20, reflux, extract, 80min under 80 DEG C of conditions, after being cooled to room temperature, centrifugal 5min under 4000r/min, takes out for subsequent use by the supernatant after centrifugal; Filtering residue 60% ethanol purge 2-3 time, then filters, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.After testing, polyphenol content is 8.91mg/g.
Embodiment 3
Aqueous enzymatic method-ethanol refluxing process extracts the method for tea-seed polyphenol, comprises the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) aqueous enzymatic extraction: take tea-seed powder 10g, be that 1:6 first adds 40ml water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, 100 DEG C are heated 5 minutes to eliminate self enzymatic activity of tea-seed powder, then room temperature is cooled to, add the enzyme liquid and residue water that have configured, stir, at 60 DEG C, enzymolysis 2h under stirring condition; Described enzyme liquid is that cellulase is dissolved in part distilled water and makes fast under cryogenic, and the consumption of cellulase is 3mg/g tea-seed powder;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 40%, solid-liquid ratio is 1:30, reflux, extract, 100min under 80 DEG C of conditions, to be cooled to after room temperature centrifugal 15min under 3000r/min, the supernatant after centrifugal is taken out for subsequent use; Filtering residue 60% ethanol purge 2-3 time, then filters, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.After testing, polyphenol content is 9.44mg/g.
Embodiment 4
Aqueous enzymatic method-ethanol refluxing process extracts the method for tea-seed polyphenol, comprises the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) aqueous enzymatic extraction: take tea-seed powder 10g, be that 1:5 first adds 30ml water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, 100 DEG C are heated 5 minutes to eliminate self enzymatic activity of tea-seed powder, then room temperature is cooled to, add the enzyme liquid and residue water that have configured, stir, at 60 DEG C, enzymolysis 1.5h under stirring condition; Described enzyme liquid is that cellulase is dissolved in part distilled water and makes fast under cryogenic, and the consumption of cellulase is 2mg/g tea-seed powder;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 50%, solid-liquid ratio is 1:25, reflux, extract, 90min under 80 DEG C of conditions, after being cooled to room temperature, centrifugal 8min under 3500r/min, takes out for subsequent use by the supernatant after centrifugal; Filtering residue 60% ethanol purge 2-3 time, then filters, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.After testing, polyphenol content is 9.41mg/g.
Claims (6)
1. aqueous enzymatic method-ethanol refluxing process extracts a method for tea-seed polyphenol, it is characterized in that: comprise the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) taking tea-seed powder, is that 1:4-6 first adds part water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, and heating enzyme denaturing, is then cooled to room temperature, adds enzyme liquid and residue water, stirs, regulate temperature 60 C, stir enzymolysis 1.0-2.0h;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 40%-60%, feed liquid weight/volume is 1:20-30, reflux, extract, 80-100min under 70-90 DEG C of condition, after being cooled to room temperature, centrifugal 5-15min under 3500-4500r/min, takes out for subsequent use by the supernatant after centrifugal; Filtering residue with 60% ethanol purge for several times, then filter, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.
2. aqueous enzymatic method as claimed in claim 1-ethanol refluxing process extracts the method for tea-seed polyphenol, it is characterized in that: in described step (2), enzyme liquid is that cellulase is dissolved in part distilled water and makes, and the consumption of cellulase is 1-3mg/g tea-seed powder; Carry out fast under cryogenic during described enzyme liquid preparation.
3. aqueous enzymatic method-ethanol refluxing process as claimed in claim 2 extracts the method for tea-seed polyphenol, it is characterized in that: the consumption of described step (2) cellulase is 2mg/g tea-seed powder.
4. aqueous enzymatic method-ethanol refluxing process as claimed in claim 1 extracts the method for tea-seed polyphenol, it is characterized in that: in described step (2), the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor is 1:5, and enzymolysis time is 1.5h.
5. aqueous enzymatic method-ethanol refluxing process as claimed in claim 1 extracts the method for tea-seed polyphenol, and it is characterized in that: in described step (3) mixed liquor, the percent by volume of ethanol is 49%, solid-liquid ratio is 1:24, and Extracting temperature is 80 DEG C.
6. the aqueous enzymatic method as described in as arbitrary in claim 1-5-ethanol refluxing process extracts the method for tea-seed polyphenol, it is characterized in that: comprise the following steps:
(1) tea-seed powder preparation: tea-seed shells, and pulverizes and sieves, and dries to constant weight;
(2) aqueous enzymatic extraction: take tea-seed powder, be that 1:5 first adds part water according to the feed liquid weight/volume of camellia oleosa seed powder and reactant liquor, 100 DEG C are heated 5 minutes to eliminate self enzymatic activity of tea-seed powder, then room temperature is cooled to, add the enzyme liquid and residue water that have configured, stir, at 60 DEG C, enzymolysis 1.5h under stirring condition; Described enzyme liquid is that cellulase is dissolved in part distilled water and makes fast under cryogenic, and the consumption of cellulase is 2mg/g tea-seed powder;
(3) alcohol reflux: add ethanol in the tea-seed powder mixed liquor after enzymolysis, the percent by volume of ethanol in mixed liquor is made to be 40%-60%, solid-liquid ratio is 1:24, reflux, extract, 90min under 80 DEG C of conditions, after being cooled to room temperature, centrifugal 10min under 3500r/min, takes out for subsequent use by the supernatant after centrifugal; Filtering residue 60% ethanol purge 2-3 time, then filters, filtrate and centrifugal after supernatant merge, rotary evaporation, be drying to obtain object product.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105125743A (en) * | 2015-09-08 | 2015-12-09 | 华侨大学 | Method of extracting combination type phenolic compounds from camellia sinensis seeds |
| CN109222096A (en) * | 2018-09-17 | 2019-01-18 | 南昌大学 | The extracting method of polyphenol is combined in carrot dietary fiber |
| CN109609256A (en) * | 2018-12-14 | 2019-04-12 | 华侨大学 | A kind of method of improved extracting tea seed oil by using enzymatic hydrolysis |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977162A (en) * | 2012-12-17 | 2013-03-20 | 贵州泰谷农业科技有限公司 | Method for producing high-purity tea saponin |
| US20130216521A1 (en) * | 2012-02-03 | 2013-08-22 | Melinda Fernyhough Culver | Dietary supplement for enhancing animal health |
-
2013
- 2013-09-23 CN CN201310435156.8A patent/CN104435363B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130216521A1 (en) * | 2012-02-03 | 2013-08-22 | Melinda Fernyhough Culver | Dietary supplement for enhancing animal health |
| CN102977162A (en) * | 2012-12-17 | 2013-03-20 | 贵州泰谷农业科技有限公司 | Method for producing high-purity tea saponin |
Non-Patent Citations (2)
| Title |
|---|
| 杨晓凤: "酶法提取老鹰茶中黄酮类化合物", 《西南农业学报》 * |
| 王华清等: "茶叶籽中总黄酮的提取及结构的初步鉴定", 《食品工业科技》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105125743A (en) * | 2015-09-08 | 2015-12-09 | 华侨大学 | Method of extracting combination type phenolic compounds from camellia sinensis seeds |
| CN109222096A (en) * | 2018-09-17 | 2019-01-18 | 南昌大学 | The extracting method of polyphenol is combined in carrot dietary fiber |
| CN109609256A (en) * | 2018-12-14 | 2019-04-12 | 华侨大学 | A kind of method of improved extracting tea seed oil by using enzymatic hydrolysis |
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| CN104435363B (en) | 2018-10-19 |
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Address after: 564100 green food industrial park, Meitan County, Guizhou, Zunyi Province, 101 floor 1, building 6, clear ecology science and Technology Park Co-patentee after: Wang Xiaoqin Patentee after: Guizhou qianchazhiguang Biotechnology Co.,Ltd. Address before: 564100 green food industrial park, Meitan County, Guizhou, Zunyi Province, 101 floor 1, building 6, clear ecology science and Technology Park Co-patentee before: Wang Xiaoqin Patentee before: GUIZHOU ZHOUYIQING BIOTECHNOLOGY CO.,LTD. |
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Effective date of registration: 20191226 Address after: 564100 green food industrial park, Meitan County, Guizhou, Zunyi Province, 101 floor 1, building 6, clear ecology science and Technology Park Patentee after: Guizhou qianchazhiguang Biotechnology Co.,Ltd. Address before: 564100 green food industrial park, Meitan County, Guizhou, Zunyi Province, 101 floor 1, building 6, clear ecology science and Technology Park Co-patentee before: Wang Xiaoqin Patentee before: Guizhou qianchazhiguang Biotechnology Co.,Ltd. |