CN103211853B - Blueberry leaves extract with polyphenol oxidase inhibitory activity - Google Patents
Blueberry leaves extract with polyphenol oxidase inhibitory activity Download PDFInfo
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- CN103211853B CN103211853B CN201310172411.4A CN201310172411A CN103211853B CN 103211853 B CN103211853 B CN 103211853B CN 201310172411 A CN201310172411 A CN 201310172411A CN 103211853 B CN103211853 B CN 103211853B
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Abstract
The invention provides a blueberry leaves extract with polyphenol oxidase inhibitory activity. The extract is obtained by decocting blueberry leaves in water, enriching the water decoction with macroporous resin, and eluting the extract without ethanol. The blueberry leaves extract can significantly inhibit the activity of polyphenol oxidase in vitro.
Description
Technical field
The present invention relates to a kind of leaf of Fructus vaccini vitis-idaeae extract with polyphenol oxidase inhibitory activity.
Background technology
Polyphenol oxidase (EC1.14.18.1), also known as tryrosinase, is extensively formed in the bodies of aminal and plant, and be the key enzyme of organism synthesis of melanin, it has single phenolase and o-diphenolase activity.Activity and the melanic generation of polyphenol oxidase have close relationship.In body, it is by TYR hydroxylation, produce ortho position dihydroxyphenylalanine (L-3,4 dihydroxyphenylalanine) (single phenolase vigor), again dopa oxidase is become DOPA quinone (diphenol enzyme activity), DOPA quinone and then generate a series of colors causing brownization, its vigor is suppressed to get final product the generation of check melanin, can be used for prevention and therapy pigmentation and melanoma disease etc., and can as skin-lightening cosmetic additive in cosmetic industry, therefore, it is necessary for developing a kind of extract with free radical scavenging activity, have broad application prospects.Polyphenol oxidase inhibitory activity Cleaning Principle: the principle of the mensuration of polyphenol oxidase inhibitory activity is that substrate for enzymatic activity L-DOPA generation dopachrome (Dopachrome) has absorption maximum at 475nm wavelength place, under inhibitor existent condition, light absorption value can decline.Enzymic catalytic reaction is carried out to inhibitor, measures light absorption value, calculate suppression ratio, its inhibitory action to polyphenol oxidase activity can be evaluated.
Summary of the invention
The invention provides a kind of leaf of Fructus vaccini vitis-idaeae extract with polyphenol oxidase inhibitory activity.
The present invention is achieved by the following technical solutions:
After Blue berry leaf is pulverized, be that solution heated and boiled extracts three times with water, after filtering, merging filtrate is also concentrated uses macroporous resin enrichment effective ingredient again, respectively with the alcoholic solution eluting of deionized water and variable concentrations, preserve after each segmentation solution solvent recovery of gained, lyophilization, the position that wherein 50% ethanol elution obtains is extract of the present invention.
Extract of the present invention is that after being pulverized by 100g Blue berry leaf, take water as solution heated and boiled, add water 1000mL at every turn and extract three times, merging filtrate after filtering, concentrated extracting solution, to 400mL, first obtains sample solution; Sample solution adds D101 macroporous resin (100g) chromatographic column, and with the alcoholic solution (each concentration 2L) of deionized water (2L) and variable concentrations, for eluant, flow velocity is about 6mL/min respectively; The position solution solvent obtained by 50% alcoholic solution eluting reclaims, and after concentrated solution lyophilization, carries out weighing and seals preservation, being extract of the present invention.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further illustrated:
Embodiment 1 one kinds has the preparation of the leaf of Fructus vaccini vitis-idaeae extract of polyphenol oxidase inhibitory activity
After being pulverized by 100g Blue berry leaf, take water as solution heated and boiled, add water 1000mL at every turn and extract three times, merging filtrate after filtering, concentrated extracting solution, to 400mL, first obtains sample solution; Sample solution adds D101 macroporous resin (100g) chromatographic column, and with the alcoholic solution (each concentration 2L) of deionized water (2L) and variable concentrations, for eluant, flow velocity is about 6mL/min respectively; The position solution solvent obtained by 50% alcoholic solution eluting reclaims, and after concentrated solution lyophilization, carries out weighing and seals preservation, being extract of the present invention.
Embodiment 2 extract is to the inhibition of polyphenol oxidase
Preparation phosphate buffer solution (0.10mol/L pH6.8), polyphenol oxidase enzymatic solution (200U/mL), levodopa solution (L-DOPA, 5mmol/L), by the 2.5mg/mL sample of itself and embodiment 1, 200U/mL polyphenol oxidase adds in 96 hole ELISA Plate according to institute's expense successively, use vibration plate device vibration 5min, sample is fully combined with polyphenol oxidase, ELISA Plate is placed in aluminum lunch box, incubation 10min in 25 DEG C of water-baths, add 5mmol/L L-DOPA again, incubation 10min at 37 DEG C, be placed in microplate reader rapidly, OD value is measured under 475nm.
Extract has inhibitory action at 1mg/ml-25mg/ml to polyphenol oxidase, and experimental data, as table 1, suppresses curve to be Y=29.073X+32.691, R2=0.9986, IC50=3.94mg/mL.
Table 1 extract suppresses the effect of polyphenol oxidase
Claims (1)
1. a leaf of Fructus vaccini vitis-idaeae extract is preparing the application with the compositions of polyphenol oxidase inhibitory activity, it is characterized in that: after 100g Blue berry leaf is pulverized, take water as solution heated and boiled, add water 1000mL at every turn and extract three times, merging filtrate after filtering, concentrated extracting solution, to 400mL, first obtains sample solution; Sample solution adds D101 macroporous resin 100g chromatographic column, respectively with each concentration 2L of alcoholic solution of deionized water 2L and variable concentrations for eluant, flow velocity is 6mL/min; The position solution solvent obtained by 50% alcoholic solution eluting reclaims, and after concentrated solution lyophilization, carries out weighing and seals preservation, being extract.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201310172411.4A CN103211853B (en) | 2013-05-09 | 2013-05-09 | Blueberry leaves extract with polyphenol oxidase inhibitory activity |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310172411.4A CN103211853B (en) | 2013-05-09 | 2013-05-09 | Blueberry leaves extract with polyphenol oxidase inhibitory activity |
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| Publication Number | Publication Date |
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| CN103211853A CN103211853A (en) | 2013-07-24 |
| CN103211853B true CN103211853B (en) | 2015-02-04 |
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| CN201310172411.4A Expired - Fee Related CN103211853B (en) | 2013-05-09 | 2013-05-09 | Blueberry leaves extract with polyphenol oxidase inhibitory activity |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103211852A (en) * | 2013-05-09 | 2013-07-24 | 南京紫玉蓝莓科技有限公司 | Blueberry leaf extractive with radical scavenging activity |
| CN105558255B (en) * | 2015-12-29 | 2019-10-01 | 陕西省植物蛋白食品工程技术研究中心有限公司 | Blueberry leaf peptide nutrient food and preparation method thereof |
| CN110760512A (en) * | 2019-04-15 | 2020-02-07 | 吉林省普蓝高科技有限公司 | Antisense nucleic acid for inhibiting activity of blueberry polyphenol oxidase and application thereof |
| CN111418752A (en) * | 2020-05-12 | 2020-07-17 | 昆明理工大学 | Method for inactivating activity of dissolved polyphenol oxidase and membrane-bound polyphenol oxidase |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101102746A (en) * | 2005-01-11 | 2008-01-09 | 丁世荣 | Skin-condition improving composition comprising vaccinium uliginosum extract and method for preparation thereof |
| CN102718738A (en) * | 2012-07-04 | 2012-10-10 | 贵阳学院 | Method for extracting, separating and purifying anthocyanin from blueberry peel and residues |
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2013
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101102746A (en) * | 2005-01-11 | 2008-01-09 | 丁世荣 | Skin-condition improving composition comprising vaccinium uliginosum extract and method for preparation thereof |
| CN102718738A (en) * | 2012-07-04 | 2012-10-10 | 贵阳学院 | Method for extracting, separating and purifying anthocyanin from blueberry peel and residues |
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| CN103211853A (en) | 2013-07-24 |
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