CN103408964A - Method for improving extraction yield of melanin in Vaccinium bracteatum Thunb by using polyphenol oxidase and tea polyphenol - Google Patents
Method for improving extraction yield of melanin in Vaccinium bracteatum Thunb by using polyphenol oxidase and tea polyphenol Download PDFInfo
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Abstract
The invention provides a method for improving extraction yield of melanin in Vaccinium bracteatum Thunb by using polyphenol oxidase and tea polyphenol, which relates to a preparation and extraction method for a natural pigment. The method comprises the following steps: with leaves of Vaccinium bracteatum Thunb as a main raw material, carrying out beating, releasing enzymes like endogenous polyphenol oxidase and peroxidase which are beneficial for formation of melanin and releasing precursor substances like phenolic compounds which form melanin at the same time; then adding exogenous polyphenol oxidase or further adding tea polyphenol to accelerate the process of enzymatic oxidation and adjust chroma so as to realize quicker obtainment of more melanin; and finally heating and extracting melanin by using an ethanol solution with a certain concentration and carrying out filtration, condensation and drying so as to obtain a melanin product. According to the invention, the method is simple; natural melanin obtained by using the method has stable quality and good security; the method enables the extraction rate of melanin to be increased by 30 to 90% compared with a water extraction method; and based on changes of the usage amount of tea polyphenol, the color of a finished pigment product covers a tonal range consisting of a blue black color, a pure black color and a reddish black color, greatly broadening the application scope of natural melanin in Vaccinium bracteatum Thunb.
Description
Technical field
The invention provides a kind of method of utilizing polyphenoloxidase and tea-polyphenol to improve melanochrome extraction efficiency yield in Fructus Vaccinii Bracteati, is a kind of preparation and extracting method of natural pigment.
Background technology
Fructus Vaccinii Bracteati belongs to Ericaceae genus vaccinium plant, and China south of the River one is with the custom that the black meal that edible its tender leaf dipping glutinous rice is cooked into is arranged among the people.Owing to can forming stable black, therefore myrtillin leave also is subject to the attention of academic and industry member.The research of the chemical composition in relevant myrtillin leave and pigment About Coloration Mechanisms of Fancy thereof has had many decades, although study to such an extent that be not very thorough, basal component, pigment synthesis approach and pigments have had some preliminary conclusions: the chemical composition contained in Vaccinium bracteatum Thunb. Leaves has hentriacontane, friedelin (ketone), epifriedelanol, β-sitosterol, Quercetin, isorientin, Hydroxycinnamic acid, meso-inositol, ursolic acid, Oleanolic Acid and abundant amino acid and organic acid etc.Wherein quercetin content is up to 3.52% of extract, and the flavones total amount accounts for more than 11.64% of extract.Myrtillin leave pigment extract all has absorption under 286nm and 309nm, have and infer to think it may is the anthocyanin class material; Also someone thinks that the main component of its pigment is cyanogen indigo-3-glucose glycosides or iridoid.More stable when the pigment extract of leaf and foodstuff additive such as NaCl, vitamins C and phenylformic acid coexist, and can generate the brown precipitation while with FeCl3, coexisting, prompting cannot be stored with ferrous products.This pigment is stable under acidic conditions, and the next black that can make of the condition of alkaline condition and heating is deepened.
The extracting method of this pigment is mainly solvent-extraction process at present.The patent " a kind of method from extraction of natural melanin myrtillin leave " (200510104210.6) of the application in 2005 such as Chen Shaojun, the natural phant myrtillin leave of take is main raw material, adopt the zymin assisted extraction such as polygalacturonase, cellulase, finally adopt the ethanolic soln extraction of natural melanin.Patent " the extraction of flavonoid monomer in oriental blueberry melanin of the application in 2006 such as Wang Li, separate, purifying and evaluation " (application number: 200610040413.8), the Vaccinium bracteatum Thunb. Leaves of take is raw material, also to adopt extraction using alcohol, melanochrome solution will be obtained after extraction using alcohol, through macroporous adsorbent resin, the flavonoid compound in melanochrome is carried out to roughing out, by polyamide column and HW-40 post, the flavonoid compound monomer is carried out to purifying again, use again High Performance Liquid Chromatography/Mass Spectrometry, nuclear magnetic resonance technique carries out Structural Identification to purified components, therefrom separate and obtained 7 kinds of flavonoid compound monomers, comprise Quercetin, chrysin, 4',5,7-trihydroxyflavone, kaempferol and luteolin, its relative content is respectively: 37.51%, 2.26%, 9.57%, 1.72% and 15.16%.The patent " a kind of method of extracting pigments from bracteatum leaves " (application number 201110264827.X) of the application in 2011 such as Liu Dongfeng, after dry myrtillin leave is pulverized, add 2-5 doubly to measure 5% acetum and soak 2-4 hour, extract 2-4 time, after centrifugation, further adopt ultrafiltration, nanofiltration, obtain concentrated solution, add water-dispersion, upper polyamide column absorption, first wash decon with water, use 5-7BV50% ethanolic soln wash-out, elutriant is concentrated again, and lyophilize obtains the myrtillin leave pigment.
Zhou Zhaoxiang etc. just reported that water is extraction agent the melanochrome of myrtillin leave is extracted as far back as 1989 in addition, obtained water miscible myrtillin leave melanochrome (food and fermentation industries, 1989(6), 54-59).Gu Wenxiu etc. (2005) " chemistry of forest product with industry " (2005,25(4), 74-78) reported and adopted 62% ethanol to obtain myrtillin leave melanochrome with ultrasonic auxiliary extraction method.
Yet Wei state China waits 2011, and (2011,4,135-138) report thinks that the oxidative fermentation process is the prerequisite that oriental blueberry melanin forms in " Food Additives Used in China "; Resin is the better selection of purifying oriental blueberry melanin; Can not measure with the content of Flavonoid substances the content of oriental blueberry melanin.Based on this thought, Wei state China waits the patent (application number 201110097948.X) of applying for " the melanic manufacture method of a kind of water miscible plant " in 2011, different with the myrtillin leave melanochrome extraction thinking of previous patent report, the method be take myrtillin leave and is extracted melanin precursor as raw material adopts water-containing organic solvent at a certain temperature, after utilizing ultra-filtration membrane to remove macromolecular components, obtain water-soluble small molecules melanin precursor composition, add polyphenol oxidase catalyzed oxidation, after ultrafiltration membrance filter, obtain again the melanochrome crude product, through macroporous resin purification, spraying drying obtains sterling again.
Although the research of domestic oriental blueberry melanin is lasting for years, but up to this point also do not form more successful industrialization, trace it to its cause is largely because the preparation method of oriental blueberry melanin goes back utmost point imperfection, Many researchers is just the separation of vat liquor evaporate to dryness, the product of gained or be medicinal extract, and color and luster and content are all unstable, perhaps product is melanin precursor such as flavonoid or other compounds, still be not enough to be directly used in dyeing, or preparation technology's flow process is very long, is difficult to industrialization.
Summary of the invention
The present invention, just in order to overcome above-mentioned deficiency, provides a kind of method of utilizing polyphenoloxidase and tea-polyphenol to improve melanochrome extraction efficiency yield in Fructus Vaccinii Bracteati.
Technical scheme of the present invention: the myrtillin leave of take is main raw material, first making beating, endogenous polyphenoloxidase and peroxidase etc. are helped to form melanic enzyme to be discharged, simultaneously the melanic precursor substances of formation such as phenolic compound are discharged, then add the external source polyphenoloxidase or further add tea-polyphenol, make the enzymatic oxidation process be accelerated and adjust colourity, more melanochrome that obtains sooner, finally adopt the heating of finite concentration ethanolic soln to extract melanochrome, through filtering, after concentrated and drying, obtain the melanochrome product, step is:
(1) making beating raw materials: get appropriate fresh or dry myrtillin leave, add the water making beating, cross the colloidal mill circular treatment, the slurry obtained is placed in not higher than under 40 ° of C temperature condition, placing 1-2 hour, promotes melanochrome to form;
(2) add tea-polyphenol and adjust product colourity: slurry is required to add tea-polyphenol according to the finished product color, higher than 40 ° of C temperature condition, is not being carried out the insulated and stirred reaction, the input amount of tea-polyphenol be in slurry total phenol quality 0 ~ 100%(W/W);
(3) enzyme-added auxiliary catalysis oxidation: will be through the slurry of pulling an oar or the mixture that added tea-polyphenol, according to the Folin-Ciocalteu method, record total phenol content in slurries and drop into polyphenoloxidase, the input amount of polyphenoloxidase be in slurries total phenol quality 1 ~ 10%(W/W), higher than 50 ° of C temperature condition, do not carrying out the insulated and stirred reaction;
(4) extraction using alcohol: according to the volume of the slurry after catalyzed oxidation, add food grade ethanol and extract, filter extracting solution, obtain the melanochrome crude product solution;
(5) drying: the melanochrome crude product solution is reclaimed to ethanol under reduced pressure, obtaining the oriental blueberry melanin powder through spraying drying.
In step (1), the fresh myrtillin leave of per kilogram adds water 1-5L making beating, if dry myrtillin leave, the dry myrtillin leave of per kilogram adds water 10-20L making beating, after making beating, slurry is through 2-3 circular treatment of colloidal mill, and the processing fineness of colloidal mill is resized to the 2-5 micron; Slurry through colloidal mill was placed 1 ~ 2 hour under 25 ~ 40 ° of C conditions, promote melanochrome to form.
In step (2), partially black-and-blue if pigment requires, the input amount of tea-polyphenol be in slurry total phenol quality 0 ~ 10%(W/W); If pigment require inclined to one side black, the input amount of tea-polyphenol be in slurry total phenol quality 10 ~ 40%(W/W); If pigment require inclined to one side reddish black, the input amount of tea-polyphenol be in slurry total phenol quality 40 ~ 100%(W/W).
In step (3), the specific activity of added polyphenoloxidase is 10000 units/more than g; The optimum temperuture of catalytic oxidation is 25 ~ 50 ° of C, and the reaction times is 2 ~ 6 hours.
In step (4), add food grade ethanol, make final alcohol concn reach 30 ~ 60%, the volume ratio of myrtillin leave slurries and ethanol extract is not less than 1:0.8 ~ 1.5; Extraction is to carry out under not higher than 60 ° of C temperature condition, and extraction time is 2-6 hour; After extracting end, adopt expression separation; It is the ethanol of 30-60% that the rear residue of separation adds concentration, and the mass volume ratio of the ethanol of residue dry substance quality and second extraction is 1:6 ~ 10; The time of extracting for the second time is 1-2 hour, and temperature is similarly not higher than 60 ° of C; United extraction liquid, filter, and obtains the melanochrome crude product solution.
In step (5), reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains the oriental blueberry melanin powder through spraying drying.The product yield accounts for the prophyll dry weight in 26.1% ~ 36%(yield with product powder dry weight and calculates), depending on the difference of extraction conditions and difference.
The present invention is based on myrtillin leave itself has the melanochrome substrate and promotes a kind of like this understanding of enzyme that melanochrome forms, endogenous enzyme and substrate-function have been proposed first to utilize, the melanic formation of recycling external source polyphenoloxidase strengthening, then utilize external source phenolic compound strengthening matrix effect and the product tone is modified a little, effectively promoted and take melanochrome output and the product stability of myrtillin leave as main material production, improved the problem that myrtillin leave medicinal extract color and luster content is unstable or technological process is long in the past, and promoted the using value of myrtillin leave melanochrome product.Up to now, not yet utilize endogenous enzyme strengthening melanochrome to form, the report that utilizes external source phenolic compound strengthening matrix effect and the product tone is modified.
Myrtillin leave powder prepared by the present invention, change based on production process tea-polyphenol add-on, can form based on myrtillin leave originally blue-black from the blue-black to the black to the product line of red-black different tones, according to the difference of application, be adapted to the needs of the foods prods of different colours demand.
Oriental blueberry melanin powder-product prepared by the inventive method, yield is high, yield 26.1% ~ 36%(yield accounts for the prophyll dry weight with product powder dry weight and calculates), depending on the difference of extraction using alcohol condition and difference.Owing in myrtillin leave melanochrome product, also containing a certain amount of polyphenols, can play anti-oxidant and remove free radical and antibiotic effect, it can be attributed to a kind of functional foodstuff tinting material, the suitable Worth Expecting of its application prospect.
The natural black pigment that utilizes the present invention to obtain, technique is simple, the constant product quality security is good, the melanochrome extraction yield improves 30-90% than water extraction, variation according to the tea-polyphenol consumption, the final finished pigmentary colours can cover blue-black, black and red-black three tone ranges, have widened well the range of application of natural oriental blueberry melanin.
Embodiment
Embodiment 1
The lyophilize myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 3.72%(moisture content is 3.55%), accurately take and get the cryodesiccated myrtillin leave of 50g and measure and add 500mL water (i.e. 10 times of water), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 2 microns), carry out further fragmentation, carry out 2 circulations, whole shattering process notices that temperature does not surpass 40 ° of C; Placed at ambient temperature 2 hours.
Toward adding specific activity in slurry, be the polyphenoloxidase 0.093g(namely 5% of 10000 units/more than g, w/w), mix, stirring reaction is 4 hours under 35 ° of C conditions.
After reaction finishes, in reacting slurry, add 95%(v/v) ethanol 750mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:1.5, alcohol concn is finally 57%, v/v), under 35 ° of C conditions, extracted 2 hours, expression separation, it is 1:7.5 that filter residue adds the mass volume ratio of the ethanol of 50% ethanolic soln 300mL(residue dry substance quality and second extraction, final alcohol concn is 53%), under 55 ° of C conditions, extracted again 1 hour, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 1325mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 13.1g through spraying drying, and the product yield is 26.1%.This product colour tone is black-and-blue.
Embodiment 2
The lyophilize myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 3.72%(moisture content is 3.55%), accurately take and get the cryodesiccated myrtillin leave of 50g and measure and add 600mL water (i.e. 12 times of water), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 3 microns), carry out further fragmentation, carry out 2 circulations, whole shattering process notices that temperature is 25 ° of C; Placed at ambient temperature 2 hours.
Toward add in slurry tea-polyphenol powder (polyphenol content 80%) 0.23g(be equivalent to total phenol content in slurry 10%), under room temperature, stir.
Toward adding specific activity in the above-mentioned slurry that is mixed with tea-polyphenol, be the polyphenoloxidase 0.02g(namely 1% of 10000 units/more than g, w/w), mix, stirring reaction is 2 hours under 50 ° of C conditions.
After reaction finishes, in reacting slurry, add 75%(v/v) ethanol 500mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:0.8, alcohol concn is finally 34%, v/v), under 50 ° of C conditions, extracted 6 hours, expression separation, filter residue adds 80% ethanolic soln 250mL, and (mass volume ratio of the ethanol of residue dry substance quality and second extraction is 1:6, final alcohol concn is 60%), under 50 ° of C conditions, extracted again 2 hours, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 1200mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 14.3g through spraying drying, and the product yield is 28.6%.This product colour tone is black-and-blue.
Embodiment 3
The lyophilize myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 3.72%(moisture content is 3.55%), accurately take and get the cryodesiccated myrtillin leave of 50g and measure and add 750mL water (i.e. 15 times of water), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 4 microns), carry out further fragmentation, carry out 3 circulations, whole shattering process temperature is 35 ° of C; Placed at ambient temperature 1 hour.
Toward add in slurry tea-polyphenol powder (polyphenol content 80%) 0.58g(be equivalent to total phenol content in slurry 25%), under room temperature, stir.
Toward adding specific activity in the above-mentioned slurry that is mixed with tea-polyphenol, be the polyphenoloxidase 0.116g(namely 5% of 10000 units/more than g, w/w), mix, stirring reaction is 6 hours under 35 ° of C conditions.
After reaction finishes, in reacting slurry, add 95%(v/v) ethanol 650mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:0.9, alcohol concn is finally 44%, v/v), under 35 ° of C conditions, extracted 2 hours, expression separation, it is 1:7.5 that filter residue adds the mass volume ratio of the ethanol of 50% ethanolic soln 300mL(residue dry substance quality and second extraction, final alcohol concn is 46%), under 60 ° of C conditions, extracted again 1 hour, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 1480mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 14.4g through spraying drying, and the product yield is 28.8%.This product colour tone is black.
Embodiment 4
The lyophilize myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 3.72%(moisture content is 3.55%), accurately take and get the cryodesiccated myrtillin leave of 50g and measure and add 900mL water (i.e. 18 times of water), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 5 microns), carry out further fragmentation, carry out 3 circulations, whole shattering process notices that temperature does not surpass 40 ° of C; Placed at ambient temperature 1.5 hours.
Toward add in slurry tea-polyphenol powder (polyphenol content 80%) 0.88g(be equivalent to total phenol content in slurry 38%), under room temperature, stir.
Toward adding specific activity in the above-mentioned slurry that is mixed with tea-polyphenol, be the polyphenoloxidase 0.128g(namely 5% of 10000 units/more than g, w/w), mix, stirring reaction is 2 hours under 25 ° of C conditions.
After reaction finishes, the volume ratio that adds dehydrated alcohol 1350mL(Fructus Vaccinii Bracteati slurries and ethanol extract in reacting slurry is 1:1.5, alcohol concn is finally 60%, v/v), under 35 ° of C conditions, extracted 2 hours, expression separation, it is 1:8.5 that filter residue adds the mass volume ratio of the ethanol of 15% ethanolic soln 350mL(residue dry substance quality and second extraction, final alcohol concn 30%), under 50 ° of C conditions, extracted again 2 hours, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 2430mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 15.8g through spraying drying, and the product yield is 31.6%.This product colour tone is black.
Embodiment 5
The lyophilize myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 3.72%(moisture content is 3.55%), accurately take and get the cryodesiccated myrtillin leave of 50g and measure and add 1000mL water (i.e. 20 times of water), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 2 microns), carry out further fragmentation, carry out 2 circulations, whole shattering process notices that temperature does not surpass 40 ° of C; Placed at ambient temperature 2 hours.
Toward add in slurry tea-polyphenol powder (polyphenol content 80%) 1.74g(be equivalent to total phenol content in slurry 75%), under room temperature, stir.
Toward adding specific activity in the above-mentioned slurry that is mixed with tea-polyphenol, be the polyphenoloxidase 0.244g(namely 7.5% of 10000 units/more than g, w/w), mix, stirring reaction is 6 hours under 50 ° of C conditions.
After reaction finishes, in reacting slurry, add 95%(v/v) ethanol 1300mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:1.3, alcohol concn is finally 53%, v/v), under 50 ° of C conditions, extracted 6 hours, expression separation, filter residue adds 30% ethanolic soln 400mL, (mass volume ratio of the ethanol of residue dry substance quality and second extraction is 1:10, final alcohol concn is 38%) under 60 ° of C conditions, extracted again 2 hours, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 2520mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 17.3g through spraying drying, and the product yield is 34.6%.This product colour tone is reddish black.
Embodiment 6
Fresh myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 0.58%(moisture content is 83.72%), accurately take and get the fresh myrtillin leave of 500g and measure and add 1000mL water (being equivalent to per kilogram raw material 2L), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 2 microns), carry out further fragmentation, carry out 2 circulations, whole shattering process notices that temperature does not surpass 40 ° of C; Placed at ambient temperature 2 hours.
Toward add in slurry tea-polyphenol powder (polyphenol content 80%) 3.6g(be equivalent to total phenol content in slurry 100%), under room temperature, stir.
Toward adding specific activity in the above-mentioned slurry that is mixed with tea-polyphenol, be the polyphenoloxidase 0.58g(namely 10% of 10000 units/more than g, w/w), mix, stirring reaction is 4 hours under 50 ° of C conditions.
After reaction finishes, in reacting slurry, add 95%(v/v) ethanol 1500mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:1, alcohol concn is finally 47%, v/v), under 50 ° of C conditions, extracted 4 hours, expression separation, it is 1:9 that filter residue adds the mass volume ratio of the ethanol of 30% ethanol 150mL(residue dry substance quality and second extraction, final alcohol concn is 37%), under 60 ° of C conditions, extracted again 2 hours, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 2620mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 7.5g through spraying drying, and the product yield is 35.1%.This product colour tone is reddish black.
Embodiment 7
Fresh myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 0.58%(moisture content is 83.72%), accurately take and get the fresh myrtillin leave of 500g and measure and add 500mL water (being equivalent to per kilogram 1L), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 2 microns), carry out further fragmentation, carry out 2 circulations, whole shattering process notices that temperature does not surpass 40 ° of C; Placed at ambient temperature 2 hours.
Toward add in slurry tea-polyphenol powder (polyphenol content 80%) 0.72g(be equivalent to total phenol content in slurry 20%), under room temperature, stir.
Toward adding specific activity in the above-mentioned slurry that is mixed with tea-polyphenol, be the polyphenoloxidase 0.18g(namely 5% of 10000 units/more than g, w/w), mix, stirring reaction is 2 hours under 35 ° of C conditions.
After reaction finishes, in reacting slurry, add 95%(v/v) ethanol 1500mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:1.5, alcohol concn is finally 57%, v/v), under 40 ° of C conditions, extracted 6 hours, expression separation, it is 1:10 that filter residue adds the mass volume ratio of the ethanol of 25% ethanol 200mL(residue dry substance quality and second extraction, final alcohol concn is 35%), under 50 ° of C conditions, extracted again 2 hours, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 2570mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 5.1g through spraying drying, and the product yield is 29.9%.This product colour tone is black.
Embodiment 8
Fresh myrtillin leave, according to the Folin-Ciocalteu method, measuring total phenol content is that the 0.58%(moisture content is 83.72%), accurately take and get the fresh myrtillin leave of 500g and measure and add 2500mL water (being equivalent to per kilogram 5L), with tissue mashing machine, smash to pieces, adopt again colloidal mill (gap is adjusted to 2 microns), carry out further fragmentation, carry out 2 circulations, whole shattering process notices that temperature does not surpass 40 ° of C; Placed at ambient temperature 2 hours.
Toward adding specific activity in slurry, be the polyphenoloxidase 0.03g(namely 1% of 10000 units/more than g, w/w), mix, stirring reaction is 6 hours under 40 ° of C conditions.
After reaction finishes, in reacting slurry, add 70%(v/v) ethanol 2400mL(Fructus Vaccinii Bracteati slurries and the volume ratio of ethanol extract be 1:0.8, alcohol concn is finally 30%, v/v), under 60 ° of C conditions, extracted 2 hours, expression separation, it is 1:6 that filter residue adds the mass volume ratio of the ethanol of dehydrated alcohol 100mL(residue dry substance quality and second extraction, final alcohol concn is 60%), under 60 ° of C conditions, extracted again 1 hour, after expression separation, united extraction liquid, filter, obtain melanochrome crude product solution 5410mL.
The melanochrome crude product solution is reclaimed to ethanol under reduced pressure, reduced pressure is 0.09MPa, 60 ° of C, and the product of sloughing after ethanol obtains oriental blueberry melanin powder 4.7g through spraying drying, and the product yield is 26.7%.This product colour tone is black-and-blue.
Claims (9)
1. one kind is utilized polyphenoloxidase and tea-polyphenol to improve the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, it is characterized in that manufacturing step is:
(1) making beating raw materials: get appropriate fresh or dry myrtillin leave, add the water making beating, cross the colloidal mill circular treatment, the slurry obtained is placed in not higher than under 40 ° of C temperature condition, placing 1-2 hour, promotes melanochrome to form;
(2) add tea-polyphenol and adjust product colourity: slurry is required to add tea-polyphenol according to the finished product color, higher than 40 ° of C temperature condition, is not being carried out the insulated and stirred reaction, the input amount of tea-polyphenol be in slurry total phenol quality 0 ~ 100%(W/W);
(3) enzyme-added auxiliary catalysis oxidation: will be through the slurry of pulling an oar or the mixture that added tea-polyphenol, according to the Folin-Ciocalteu method, record total phenol content in slurries and drop into polyphenoloxidase, the input amount of polyphenoloxidase be in slurries total phenol quality 1 ~ 10%(W/W), higher than 50 ° of C temperature condition, do not carrying out the insulated and stirred reaction;
(4) extraction using alcohol: according to the volume of the slurry after catalyzed oxidation, add food grade ethanol and extract, filter extracting solution, obtain the melanochrome crude product solution;
(5) drying: the melanochrome crude product solution is reclaimed to ethanol under reduced pressure, obtaining the oriental blueberry melanin powder through spraying drying.
2. polyphenoloxidase and the tea-polyphenol of utilizing according to claim 1 improves the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, it is characterized in that in step (1), the fresh myrtillin leave of per kilogram adds water 1-5L making beating or the dry myrtillin leave of per kilogram adds water 10-20L making beating.
3. polyphenoloxidase and the tea-polyphenol of utilizing according to claim 2 improves the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, it is characterized in that in step (1), after making beating, slurry is through 2-3 circular treatment of colloidal mill, the processing fineness of colloidal mill is resized to the 2-5 micron.
4. the method for utilizing polyphenoloxidase and tea-polyphenol to improve melanochrome extraction yield in Fructus Vaccinii Bracteati according to claim 1, is characterized in that in step (1), the slurry through colloidal mill was placed 1 ~ 2 hour under 25 ~ 40 ° of C conditions, promotes melanochrome to form.
5. polyphenoloxidase and the tea-polyphenol of utilizing according to claim 1 improves the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, it is characterized in that in step (2), it is partially black-and-blue that pigment requires, the input amount of tea-polyphenol be in slurry total phenol quality 0 ~ 10%(W/W); Pigment require inclined to one side black, the input amount of tea-polyphenol be in slurry total phenol quality 10 ~ 40%(W/W); Pigment require inclined to one side reddish black, the input amount of tea-polyphenol be in slurry total phenol quality 40 ~ 100%(W/W).
6. polyphenoloxidase and the tea-polyphenol of utilizing according to claim 1 improves the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, it is characterized in that in step (3), and the specific activity of added polyphenoloxidase is 10000 units/more than g.
7. polyphenoloxidase and the tea-polyphenol of utilizing according to claim 1 improves the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, and the temperature that it is characterized in that catalytic oxidation is 25 ~ 50 ° of C, and the reaction times is 2 ~ 6 hours.
8. polyphenoloxidase and the tea-polyphenol of utilizing according to claim 1 improves the method that melanochrome in Fructus Vaccinii Bracteati extracts yield, it is characterized in that in step (4), add food grade ethanol, make final alcohol concn reach 30 ~ 60%, the volume ratio of myrtillin leave slurries and ethanol extract is not less than 1:0.8 ~ 1.5, extraction is to carry out under not higher than 60 ° of C temperature condition, and extraction time is 2-6 hour; After extracting end, adopt expression separation; It is the ethanol of 30-60% that the rear residue of separation adds concentration, and the mass volume ratio of the ethanol of residue dry substance quality and second extraction is 1:6 ~ 10, and the time of extracting for the second time is 1-2 hour, and temperature is similarly not higher than 60 ° of C; United extraction liquid, filter, and obtains the melanochrome crude product solution.
9. the method for utilizing polyphenoloxidase and tea-polyphenol to improve melanochrome extraction yield in Fructus Vaccinii Bracteati according to claim 1, is characterized in that in step (5), reduced pressure is 0.09MPa, 60 ° of C.
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| CN114921111A (en) * | 2022-06-02 | 2022-08-19 | 苏州大学 | Preparation process and application of blueberry pigment |
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