CN103520372B - A kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice - Google Patents
A kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice Download PDFInfo
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- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 34
- 235000009566 rice Nutrition 0.000 title claims abstract description 34
- 101710147108 Tyrosinase inhibitor Proteins 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 21
- 240000007594 Oryza sativa Species 0.000 title 1
- 241000209094 Oryza Species 0.000 claims abstract description 33
- 239000000284 extract Substances 0.000 claims abstract description 12
- 238000000605 extraction Methods 0.000 claims abstract description 10
- 239000000872 buffer Substances 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract 8
- 238000010828 elution Methods 0.000 claims abstract 7
- 238000001914 filtration Methods 0.000 claims abstract 5
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims abstract 3
- 230000001476 alcoholic effect Effects 0.000 claims abstract 2
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- 238000005238 degreasing Methods 0.000 claims description 2
- CSVGEMRSDNSWRF-UHFFFAOYSA-L disodium;dihydrogen phosphate Chemical group [Na+].[Na+].OP(O)([O-])=O.OP(O)([O-])=O CSVGEMRSDNSWRF-UHFFFAOYSA-L 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims 4
- 238000004440 column chromatography Methods 0.000 claims 1
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- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 8
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- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 102000003425 Tyrosinase Human genes 0.000 description 2
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- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- VIYKYVYAKVNDPS-HKGPVOKGSA-N (2s)-2-azanyl-3-[3,4-bis(oxidanyl)phenyl]propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1.OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 VIYKYVYAKVNDPS-HKGPVOKGSA-N 0.000 description 1
- 239000005541 ACE inhibitor Substances 0.000 description 1
- 229940086440 Angiotensin I converting enzyme inhibitor Drugs 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- AHMIDUVKSGCHAU-UHFFFAOYSA-N Dopaquinone Natural products OC(=O)C(N)CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 1
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- AHMIDUVKSGCHAU-LURJTMIESA-N L-dopaquinone Chemical compound [O-]C(=O)[C@@H]([NH3+])CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-LURJTMIESA-N 0.000 description 1
- 108091000117 Tyrosine 3-Monooxygenase Proteins 0.000 description 1
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Abstract
The present invention relates to the extracting method of tyrosinase inhibitor, aim to provide a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice.The method comprises the following steps: carry out defat by after the pretreatment of the tea-seed dregs of rice, then adds ethanol water and extracts, centrifugally obtain filtering residue and extracting solution; In filtering residue, add buffer, carry out lixiviate, supersound extraction, centrifugally obtain extracting solution; By after the extracting solution vacuum concentration that obtains before, extraction obtains raffinate, the extracting solution obtained afterwards is carried out gradient elution with 60% alcoholic solution and obtains 60% ethanol elution; By raffinate and the mixing of 60% ethanol elution, concentrated postlyophilization obtains tyrosinase inhibitor.The present invention effectively improves the extraction precision of object, substantially increase the efficiency of extraction, effectively can improve the economic benefit of tea-seed dregs of rice resource, and the tyrosinase inhibitor of preparation is a kind of tyrosinase inhibitor of efficiency natural, remarkable to the inhibition of tryrosinase.
Description
Technical field
The invention relates to the extracting method of tyrosinase inhibitor, particularly a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice.
Background technology
All kinds of mottles that melanin deposition is formed cause the puzzlement of physiology and psychology to modern especially women, and speckle dispelling has been the primary goal of numerous women beauty treatment.Research shows, tryrosinase is the key enzyme that melanin is formed.Tryrosinase is the copper desmoenzyme that a class is extensively present in organism, by by tyrosine hydroxylase, produces L-DOPA (L-DOPA), and then L-DOPA is oxidized to DOPA quinone further, thus form melanin.Meanwhile, play key player in the brown stain that tryrosinase produces in fruit and vegerable storage and transport process, outward appearance and the quality of fruit and vegerable can be reduced, thus reduce it and be worth.Therefore, can not only effectively be formed by check melanin the suppression of tryrosinase, thus the formation of all kinds of mottle of retardance human body, and can effectively prevent fruit and vegerable brown stain, keep its quality and value.
The tea-seed dregs of rice are the solid residues after tea-seed oil expression.List tea-seed oil in new food resource from December, 2009 Ministry of Public Health approval, tea-seed oil production scale constantly expands, and causes the output of this processing byproduct of the tea-seed dregs of rice also constantly to increase.Research shows, containing comprising the multiple bioactive ingredients such as polyphenol, polysaccharide, saponin and albumen in the tea-seed dregs of rice, has high Development volue.But at present except a small amount of tea-seed dregs of rice are utilized as fertilizer low value, these living resources are not also by effective exploitation.In recent years, from plant, extract safe and effective native tyrosine enzyme inhibitor to receive much concern.Yet there are no from the tea-seed dregs of rice, extract tyrosinase inhibitor relevant report or patent.
Summary of the invention
Main purpose of the present invention is to overcome deficiency of the prior art, provides one effectively can utilize tea-seed dregs of rice resource, and can extract the method for native tyrosine enzyme inhibitor.For solving the problems of the technologies described above, solution of the present invention is:
A kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice is provided, comprises the pulverizing tea seed dregs of rice, comprise following concrete steps:
Steps A: carry out defat by after the pretreatment of the tea-seed dregs of rice, with the ratio that the ethanol water of the tea-seed dregs of rice and volumetric concentration 95% is 1g:15ml ~ 1g:25ml, the ethanol water of volumetric concentration 95% is added in the tea-seed dregs of rice after defat, 30 ~ 40min is extracted at 40 ~ 50 DEG C, then after centrifugal 20 minutes, filtering residue and extracting solution is obtained with 12000 revs/min;
Step B: the ratio that the filtering residue obtained with steps A and buffer are 1g:20ml ~ 1g:30ml, buffer is added, lixiviate 2 ~ 3h at 40 ~ 60 DEG C in the filtering residue that steps A obtains, then after supersound extraction 10 ~ 20min, centrifugal 20 minutes with 12000 revs/min, obtain extracting solution; Described buffer is PH is 7 ~ 9, and containing accounting for the buffer of alkaline protease of tea-seed dregs of rice dry weight 3 ~ 5% after defat;
Step C: extracting solution steps A obtained is concentrated in vacuo to 1/5 ~ 1/15 of original volume, then with the ratio that the volume ratio of the extracting solution after concentrated and chloroform is 1:1, adds chloroform and extracts, obtain raffinate in the extracting solution after concentrated;
Step D: the extracting solution obtained by step B is concentrated in vacuo to 1/5 ~ 1/7 of original volume, cross D101 macroporous resin and carry out column chromatography, the alcoholic solution being respectively 20%, 60% and 100% by volumetric concentration carries out gradient elution, and elution flow rate is 1ml/min, collects 60% ethanol elution;
Step e: by the 60% ethanol elution mixing obtained in the raffinate obtained in step C and step D, concentrated postlyophilization, namely obtains extracting tyrosinase inhibitor from the tea-seed dregs of rice.
As further improvement, the pretreatment in described steps A is pulverized by the tea-seed dregs of rice, then crosses 40 ~ 60 mesh sieves.
As further improvement, degreasing method in described steps A is: be the ratio of 1g:10ml ~ 1g:20ml according to the pretreated tea-seed dregs of rice and normal hexane, normal hexane is added in the pretreated tea-seed dregs of rice, lixiviate 1 ~ 2h at 30 ~ 50 DEG C, then abandon supernatant with 5000 revs/min after centrifugal 15 minutes, get residue and dry.
As further improvement, the vigor of the alkaline protease in described step B is not less than 20000U/g.
As further improvement, the buffer in described step B is sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution.
As further improvement, the frequency of the supersound extraction in described step B is 150 ~ 250W.
Compared with prior art, the invention has the beneficial effects as follows:
The present invention gets the extraction precision that effectively improve object indescribably by doping region, and is substantially increased the efficiency of extraction in conjunction with supersound extraction by enzyme process, is applicable to industrialization and produces.Meanwhile, the tyrosinase inhibitor of a kind of efficiency natural of the tyrosinase inhibitor prepared in the present invention, remarkable to the inhibition of tryrosinase.In addition, the technology of the present invention effectively can improve the economic benefit of tea-seed dregs of rice resource.
Accompanying drawing explanation
Fig. 1 is that the tyrosinase inhibitor that extracts in the embodiment of the present invention is to the inhibition comparison diagram of tryrosinase.
Detailed description of the invention
Below in conjunction with accompanying drawing and detailed description of the invention, the present invention is described in further detail:
The method concrete steps that the present invention extracts angiotensin I-converting enzyme inhibitor from tea peel comprise:
Steps A: by the pretreatment of the tea-seed dregs of rice, preprocess method is pulverized by the tea-seed dregs of rice, then crosses 40 ~ 60 mesh sieves; Defat is carried out after the pretreatment of the tea-seed dregs of rice, degreasing method is: be the ratio of 1g:10ml ~ 1g:20ml according to the pretreated tea-seed dregs of rice and normal hexane, normal hexane is added in the pretreated tea-seed dregs of rice, lixiviate 1 ~ 2h at 30 ~ 50 DEG C, then abandon supernatant with 5000 revs/min after centrifugal 15 minutes, get residue and dry.With the ratio that the ethanol water of the tea-seed dregs of rice and volumetric concentration 95% is 1g:15ml ~ 1g:25ml, the ethanol water of volumetric concentration 95% is added in the tea-seed dregs of rice after defat, 30 ~ 40min is extracted at 40 ~ 50 DEG C, then after centrifugal 20 minutes, filtering residue and extracting solution is obtained with 12000 revs/min.
Step B: the ratio that the filtering residue obtained with steps A and buffer are 1g:20ml ~ 1g:30ml, buffer is added in the filtering residue that steps A obtains, lixiviate 2 ~ 3h at 40 ~ 60 DEG C, be after the supersound extraction 10 ~ 20min of 150 ~ 250W again by frequency, centrifugal 20 minutes with 12000 revs/min, obtain extracting solution.Described buffer is PH is 7 ~ 9, and containing accounting for the buffer of alkaline protease of tea-seed dregs of rice dry weight 3 ~ 5% after defat, the vigor of described alkaline protease is not less than 20000U/g, buffer preferably phosphoric acid disodium hydrogen-phosphate sodium dihydrogen buffer solution.
Step C: extracting solution steps A obtained is concentrated in vacuo to 1/5 ~ 1/15 of original volume, then with the ratio that the volume ratio of the extracting solution after concentrated and chloroform is 1:1, adds chloroform and extracts, obtain raffinate in the extracting solution after concentrated;
Step D: the extracting solution obtained by step B is concentrated in vacuo to 1/5 ~ 1/7 of original volume, cross D101 macroporous resin and carry out column chromatography, the alcoholic solution being respectively 20%, 60% and 100% by volumetric concentration carries out gradient elution, and elution flow rate is 1ml/min, collects 60% ethanol elution;
Step e: by the 60% ethanol elution mixing obtained in the raffinate obtained in step C and step D, concentrated postlyophilization, namely obtains extracting tyrosinase inhibitor from the tea-seed dregs of rice.
The following examples can make professional and technical personnel's comprehend the present invention of this specialty, but do not limit the present invention in any way.
Embodiment 1
Steps A: by the pretreatment of the tea-seed dregs of rice, preprocess method is pulverized by the tea-seed dregs of rice, then crosses 40 mesh sieves; Defat is carried out after the pretreatment of the tea-seed dregs of rice, degreasing method is: be the ratio of 1g:10ml according to the pretreated tea-seed dregs of rice and normal hexane, normal hexane is added in the pretreated tea-seed dregs of rice, lixiviate 2h at 30 DEG C, then 5000 revs/min, abandon supernatant after centrifugal 15 minutes, get residue and dry; With the ratio that the ethanol of the tea-seed dregs of rice and volumetric concentration 95% is 1g:15mlml, in the tea-seed dregs of rice after 20g defat, add the ethanol of volumetric concentration 95%, at 40 DEG C, extract 40min, then 12000 revs/min, after centrifugal 20 minutes, obtain filtering residue and extracting solution;
Step B: the ratio that the filtering residue obtained with steps A and sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution are 1g:20ml, sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution is added in the filtering residue that steps A obtains, lixiviate 3h at 40 DEG C, be after the supersound extraction 20min of 150W again by frequency, 12000 revs/min, centrifugal 20 minutes, obtain extracting solution; Described sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution is PH is 7, and containing accounting for the buffer of alkaline protease of tea-seed dregs of rice dry weight 3% after defat, the vigor of described alkaline protease is not less than 20000U/g;
Step C: extracting solution steps A obtained is concentrated in vacuo to 1/5 of original volume, then with the ratio that the volume ratio of the extracting solution after concentrated and chloroform is 1:1, adds chloroform and extracts, obtain raffinate in the extracting solution after concentrated;
Step D: the extracting solution obtained by step B is concentrated in vacuo to 1/5 of original volume, cross D101 macroporous resin and carry out column chromatography, the alcoholic solution being respectively 20%, 60% and 100% by volumetric concentration carries out gradient elution, and elution flow rate is 1ml/min, collects 60% ethanol elution;
Step e: by the 60% ethanol elution mixing obtained in the raffinate obtained in step C and step D, concentrated postlyophilization, namely obtains extracting tyrosinase inhibitor 0.72g from the tea-seed dregs of rice, yield is that 3.6%(accounts for tea-seed dregs of rice dry weight after defat).
Embodiment 2
Steps A: by the pretreatment of the tea-seed dregs of rice, preprocess method is pulverized by the tea-seed dregs of rice, then crosses 50 mesh sieves; Defat is carried out after the pretreatment of the tea-seed dregs of rice, degreasing method is: be the ratio of 1g:15ml according to the pretreated tea-seed dregs of rice and normal hexane, normal hexane is added in the pretreated tea-seed dregs of rice, lixiviate 1.5h at 40 DEG C, then 5000 revs/min, abandon supernatant after centrifugal 15 minutes, get residue and dry; With the ratio that the ethanol of the tea-seed dregs of rice and volumetric concentration 95% is 1g:20ml, in the tea-seed dregs of rice after 20g defat, add the ethanol of volumetric concentration 95%, at 45 DEG C, extract 35min, then 12000 revs/min, after centrifugal 20 minutes, obtain filtering residue and extracting solution;
Step B: the ratio that the filtering residue obtained with steps A and sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution are 1g:25ml, sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution is added in the filtering residue that steps A obtains, lixiviate 2.5h at 50 DEG C, be after the supersound extraction 15min of 200W again by frequency, 12000 revs/min, centrifugal 20 minutes, obtain extracting solution; Described sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution is PH is 8, and containing accounting for the buffer of alkaline protease of tea-seed dregs of rice dry weight 4% after defat, the vigor of described alkaline protease is not less than 20000U/g;
Step C: extracting solution steps A obtained is concentrated in vacuo to 1/10 of original volume, then with the ratio that the volume ratio of the extracting solution after concentrated and chloroform is 1:1, adds chloroform and extracts, obtain raffinate in the extracting solution after concentrated;
Step D: the extracting solution obtained by step B is concentrated in vacuo to 1/6 of original volume, cross D101 macroporous resin and carry out column chromatography, the alcoholic solution being respectively 20%, 60% and 100% by volumetric concentration carries out gradient elution, and elution flow rate is 1ml/min, collects 60% ethanol elution;
Step e: by the 60% ethanol elution mixing obtained in the raffinate obtained in step C and step D, concentrated postlyophilization, namely obtains extracting tyrosinase inhibitor 0.86g from the tea-seed dregs of rice, yield is that 4.3%(accounts for tea-seed dregs of rice dry weight after defat).
Embodiment 3
Steps A: by the pretreatment of the tea-seed dregs of rice, preprocess method is pulverized by the tea-seed dregs of rice, then crosses 60 mesh sieves; Defat is carried out after the pretreatment of the tea-seed dregs of rice, degreasing method is: be the ratio of 1g:20ml according to the pretreated tea-seed dregs of rice and normal hexane, normal hexane is added in the pretreated tea-seed dregs of rice, lixiviate 1h at 50 DEG C, then 5000 revs/min, abandon supernatant after centrifugal 15 minutes, get residue and dry; With the ratio that the ethanol of the tea-seed dregs of rice and volumetric concentration 95% is 1g:25ml, in the tea-seed dregs of rice after 20g defat, add the ethanol of volumetric concentration 95%, at 50 DEG C, extract 30min, then 12000 revs/min, after centrifugal 20 minutes, obtain filtering residue and extracting solution;
Step B: the ratio that the filtering residue obtained with steps A and sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution are 1g:30ml, sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution is added in the filtering residue that steps A obtains, lixiviate 2h at 60 DEG C, be after the supersound extraction 10min of 250W again by frequency, 12000 revs/min, centrifugal 20 minutes, obtain extracting solution; Described sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution is PH is 9, and containing accounting for the buffer of alkaline protease of tea-seed dregs of rice dry weight 5% after defat, the vigor of described alkaline protease is not less than 20000U/g;
Step C: extracting solution steps A obtained is concentrated in vacuo to 1/15 of original volume, then with the ratio that the volume ratio of the extracting solution after concentrated and chloroform is 1:1, adds chloroform and extracts, obtain raffinate in the extracting solution after concentrated;
Step D: the extracting solution obtained by step B is concentrated in vacuo to 1/7 of original volume, cross D101 macroporous resin and carry out column chromatography, the alcoholic solution being respectively 20%, 60% and 100% by volumetric concentration carries out gradient elution, and elution flow rate is 1ml/min, collects 60% ethanol elution;
Step e: by the 60% ethanol elution mixing obtained in the raffinate obtained in step C and step D, concentrated postlyophilization, namely obtains extracting tyrosinase inhibitor 0.98g from the tea-seed dregs of rice, yield is that 4.9%(accounts for tea-seed dregs of rice dry weight after defat).
The tyrosinase inhibitor of above-mentioned 3 embodiments is carried out the checking of tryrosinase inhibition: tyrosinase inhibitor will be extracted from the tea-seed dregs of rice and be dissolved in distilled water, get the above-mentioned solution of 100 μ l, add the tyrosine solution 94 μ l that PH is phosphate buffer 56 μ l and the 2.0mmol/L of 6.8, to be mixed evenly after, tyrosinase solution 50 μ l (50U) is added rapidly in ice bath, incubation 10min in 37 DEG C, measures absorbance in 490nm place.
Tyrosinase inhibition rate is calculated as follows:
Tyrosinase inhibition rate (%)=[(a-b)-(c-d)]/[a-b] × 100%
A: do not add extract and the absorbance that enzyme-added mixed liquor is surveyed;
B: do not add the absorbance that the also not enzyme-added mixed liquor of extract is surveyed;
C: the absorbance that the mixed liquor adding extract and enzyme is surveyed;
D: add extract and the absorbance that not enzyme-added mixed liquor is surveyed.
As shown in Figure 1, as can be seen from experimental data, the tyrosinase inhibitor that application the technology of the present invention is extracted from the tea-seed dregs of rice has significant inhibitory action to tryrosinase, and present dose-dependent effect, the medicines such as beauty treatment and eliminating spot and health product exploitation can be applied to, and the field such as fruit and vegerable keeping fresh and protecting color.
Claims (6)
1. from the tea-seed dregs of rice, extract a method for tyrosinase inhibitor, comprise the pulverizing tea seed dregs of rice, it is characterized in that, the described method extracting tyrosinase inhibitor from the tea-seed dregs of rice specifically comprises the following steps:
Steps A: carry out defat by after the pretreatment of the tea-seed dregs of rice, with the ratio that the ethanol water of the tea-seed dregs of rice and volumetric concentration 95% is 1g:15ml ~ 1g:25ml, the ethanol water of volumetric concentration 95% is added in the tea-seed dregs of rice after defat, 30 ~ 40min is extracted at 40 ~ 50 DEG C, then after centrifugal 20 minutes, filtering residue and extracting solution is obtained with 12000 revs/min;
Step B: the ratio that the filtering residue obtained with steps A and buffer are 1g:20ml ~ 1g:30ml, buffer is added, lixiviate 2 ~ 3h at 40 ~ 60 DEG C in the filtering residue that steps A obtains, then after supersound extraction 10 ~ 20min, centrifugal 20 minutes with 12000 revs/min, obtain extracting solution; Described buffer is PH is 7 ~ 9, and containing accounting for the buffer of alkaline protease of tea-seed dregs of rice dry weight 3 ~ 5% after defat;
Step C: extracting solution steps A obtained is concentrated in vacuo to 1/5 ~ 1/15 of original volume, then with the ratio that the volume ratio of the extracting solution after concentrated and chloroform is 1:1, adds chloroform and extracts, obtain raffinate in the extracting solution after concentrated;
Step D: the extracting solution obtained by step B is concentrated in vacuo to 1/5 ~ 1/7 of original volume, cross D101 macroporous resin and carry out column chromatography, the alcoholic solution being respectively 20%, 60% and 100% by volumetric concentration carries out gradient elution, and elution flow rate is 1ml/min, collects 60% ethanol elution;
Step e: by the 60% ethanol elution mixing obtained in the raffinate obtained in step C and step D, concentrated postlyophilization, namely obtains extracting tyrosinase inhibitor from the tea-seed dregs of rice.
2. a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice according to claim 1, is characterized in that, the pretreatment in described steps A is pulverized by the tea-seed dregs of rice, then crosses 40 ~ 60 mesh sieves.
3. a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice according to claim 1, it is characterized in that, degreasing method in described steps A is: be the ratio of 1g:10ml ~ 1g:20ml according to the pretreated tea-seed dregs of rice and normal hexane, normal hexane is added in the pretreated tea-seed dregs of rice, lixiviate 1 ~ 2h at 30 ~ 50 DEG C, then abandon supernatant with 5000 revs/min after centrifugal 15 minutes, get residue and dry.
4. a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice according to claim 1, it is characterized in that, the vigor of the alkaline protease in described step B is not less than 20000U/g.
5. a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice according to claim 1, is characterized in that, the buffer in described step B is sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution.
6. a kind of method extracting tyrosinase inhibitor from the tea-seed dregs of rice according to claim 1, is characterized in that, the frequency of the supersound extraction in described step B is 150 ~ 250W.
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| CN102372761A (en) * | 2011-10-10 | 2012-03-14 | 安徽农业大学 | Method for extracting tea saponin from sasanglla cake |
| CN102860925A (en) * | 2012-08-30 | 2013-01-09 | 扬州泊瑞生物农业科技开发有限公司 | Application of theaflavin to preparation of tyrosinase inhibitor |
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| CN102372761A (en) * | 2011-10-10 | 2012-03-14 | 安徽农业大学 | Method for extracting tea saponin from sasanglla cake |
| CN102860925A (en) * | 2012-08-30 | 2013-01-09 | 扬州泊瑞生物农业科技开发有限公司 | Application of theaflavin to preparation of tyrosinase inhibitor |
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| 油茶籽粕中茶皂素的提取工艺研究;李敏等;《中国粮油学报》;20110531;第26卷(第5期);第38-41页，第46页 * |
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