CN103773823B - The preparation method of gardenia blue - Google Patents
The preparation method of gardenia blue Download PDFInfo
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- CN103773823B CN103773823B CN201210404931.9A CN201210404931A CN103773823B CN 103773823 B CN103773823 B CN 103773823B CN 201210404931 A CN201210404931 A CN 201210404931A CN 103773823 B CN103773823 B CN 103773823B
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Abstract
The invention discloses a kind of preparation method of gardenia blue, the method successively through enzymolysis jasminoidin, colour developing, enriching and purifying, concentrated, drying step prepare simultaneously look valency 20 ~ 40 gardenia blue and the gardenia blue of look valency more than 60.The inventive method easy handling, good stability, the high price gardenia blue productive rate prepared is high, has broad application prospects.
Description
Technical field
The invention belongs to field of natural pigment, be specifically related to a kind of preparation method of gardenia blue.
Background technology
In recent years, natural pigment obtains very fast development.But the most of natural pigment produced is all based on red, yellow two class colors.And in red, yellow, primary colors, the turnout of cyanine is less.Lack cyanine, just can not allocate green, purple, the color such as brown.Due to the existence of demand, cyanine has good market outlook.
Gardenia blue, English name (Gardeniablue), by the colour generation glycoside body of madder wort cape jasmine fruit with warm water extraction gained and the mixture (amino acids) of protein decomposition product, add beta-glucosidase, form through fermentation, deactivation, filtration, evaporation, refining, drying.In blue.Have data in literature to show, the consumption of Japan's gardenia blue in 2006 is about 100 tons, and the population that China of comparing is a large amount of, along with the continuous lifting of standard of living, the actual demand amount of China should be far above 100 tons/year.At present on the market main product to be look valency be 40 gardenia blue product, per kilogram price is between 1000 ~ 1200 yuan, and the price of raw water cape jasmine is 10 ~ 15 yuan/kilogram.
In the prior art, be in the Chinese invention patent of 200710045237.1 at application number, disclose and be obtained by reacting genipin with beta-glucoside enzyme catalysis jasminoidin, and genipin and amino acid are reacted to obtain the technology of natural gardenia blue pigment; Be in the Chinese invention patent of 200910114410.8 at application number, disclose with cape jasmine fruit powder, through extracting, filtering, concentrated, upper macroporous resin column, wash-out, alumina adsorption post separation, cellulase hydrolysis catalysis, the process such as acid hydrolysis, obtain the technology of gardenia blue pigment; Disclose the waste liquid after extracting with Gardenia Yellow for raw material at paper " two-step approach produces the research of gardenia blue pigment processing condition ", obtained the technology of gardenia blue pigment by the two-step approach of beta-glucoside enzymic fermentation and enzymatic reaction being separated.There is step complexity in existing disclosed gardenia blue preparation technology, the growing amount of gardenia blue finished product is low, the problem that cost is high, and finished product look valency higher than 100 rarely have report.
Summary of the invention
The object of the invention is to overcome above-mentioned weak point provides a kind of yield high, and can produce the preparation method of the gardenia blue of High color values and low look valency simultaneously.Extensively, cost is low, and the product prepared has taken into account the demand of high-end market and low-end market for the inventive method and raw material sources.
The object of the invention is to realize in the following manner:
A preparation method for gardenia blue, is characterized in that the method comprises the following steps:
A) enzymolysis jasminoidin: take water as solvent, adds jasminoidin, complex cellulase, anhydrous sodium acetate and Glacial acetic acid, stirring reaction 12 ~ 36 hours under 15 ~ 70 DEG C of conditions; Wherein, complex cellulase and jasminoidin mass ratio 0.5 ~ 1.5:0.5 ~ 1.5, anhydrous sodium acetate and jasminoidin mass ratio 0.3 ~ 0.5:0.6 ~ 1.2, Glacial acetic acid and jasminoidin mass ratio 0.1 ~ 0.4:0.8 ~ 1.2; By the solution centrifugal obtained after enzymolysis, discard precipitation, collect supernatant liquor;
B) develop the color: supernatant liquor is added glycine according to the ratio that jasminoidin and glycine mol ratio are 0.5 ~ 1.5:0.5 ~ 1.5 and develops the color; By the solution centrifugal obtained after colour developing, discard precipitation, collect upper solution, obtain extracting solution;
C) enriching and purifying: adopting macroporous adsorptive resins or hydrogen bonding adsorption resin post enriching and purifying, after washing 2 ~ 6 times of column volumes, is that 2 ~ 6 times of column volumes washed by 50% ~ 70% ethanol by concentration, collects water elution liquid and ethanol eluate respectively; Water elution liquid concentrates, spraying dry obtain look valency 20 ~ 40 gardenia blue, ethanol eluate concentrates, spraying dry obtains the gardenia blue of look valency more than 60.The gardenia blue of look valency of the present invention more than 60 is mainly the gardenia blue of color range 60 ~ 120.
The enzyme adopted in enzymolysis jasminoidin step described in the method is the complex cellulase of 160 ~ 2,000,000 unit of activity.Described water consumption is every 1kg jasminoidin water 20 ~ 30L; Centrifugal condition is 10000 ~ 18000rmin
-1centrifugal 15 ~ 25min under condition.Described color condition is develop the color 1 ~ 3 hour at 20 ~ 100 DEG C.The macroporous adsorbent resin that enriching and purifying adopts is polarity or nonpolar macroporous adsorption resin, and the macroporous adsorbent resin preferably adopted is HPD600 type macroporous adsorbent resin, and hydrogen bonding adsorption resin is HPD417 type polymeric adsorbent.The elution speed adopted in enriching and purifying step is 1 ~ 3BV/h, and applied sample amount is that every 1.5ml column volume adds 0.5 ~ 1ml extracting solution.In enriching and purifying step, the concentrated temperature adopted is 15 ~ 70 DEG C.
The condition that spraying dry described in the inventive method adopts is: inlet temperature is 170 ~ 180 DEG C, and the water elution liquid temperature out after concentrated is 85 ~ 90 DEG C, and the ethanol eluate temperature out after concentrated is 75 ~ 80 DEG C.
Part Experiment example is below adopted to be further expalined the present invention:
One, in color reaction, different aminoacids used becomes the impact of chromatic effect for gardenia blue entirety.
The present invention has carried out screening experiment to amino acid such as L-glutamic acid, glycine, leucines, and result shows, and under 590nm absorbing wavelength, the absorbancy of reaction later stage solution is higher, and gardenia blue component content is larger.Can find out that colour developing is after 8 hours clearly by Fig. 1, use glycine colour developing to develop the color with use L-glutamic acid and there is notable difference, because the growing amount of gardenia blue is different, this is just related to the height of yield, so this process selection glycine can significantly improve must measuring of gardenia blue as developer, sample-pretreating method is identical, and relative yield increased value can use absorbance ratio to ask for.Develop the color after 8 hours, relative to L-glutamic acid, select glycine, absorbancy improves about 33%, and look valency obviously increases.
Two, gardenia blue enriching and purifying resin column and elutriant screening experiment
Through a large amount of preliminary experiments, further screening experiment is carried out according to the operation steps of embodiment 1 to the HPD100 macroporous adsorptive resins chosen, HPD400A macroporous adsorptive resins, ADS-17 macroporous adsorptive resins, HPD600 macroporous adsorptive resins, HPD417 hydrogen bonding adsorption resin post, gardenia blue effective yield (i.e. the rate of extract, the calculating according to look valency more than 40) and the look valency of result display HPD600 macroporous adsorptive resins, the acquisition of HPD417 hydrogen bonding adsorption resin post are higher.The results are shown in Table 1.
Table 1
Three, the drying process screening experiment of gardenia blue washing lotion after resin is crossed.
Under the identical condition of stock liquid, select 100 DEG C of drying with water baths, rotary evaporation to concentrate+drying under reduced pressure, rotary evaporation concentrates+vacuum freezedrying, spraying dry, result shows: spray-dired effect is optimum, show that the look valency of product is the highest.
The results are shown in Table 2:
Table 2
Beneficial effect of the present invention compared with the prior art:
1, the enzyme price of hydrolysis jasminoidin is low, hydrolysis result good.
Catalyzer of the present invention is complex cellulase, and the effect of its enzymolysis jasminoidin can be equal to or higher than beta-glucosidase conventional in prior art, but price reduces greatly.Refer to table 3.
Table 3
2, adopt the inventive method, 1kg jasminoidin raw material prepares look valency more than 60, particularly color range be the yield of the gardenia blue of 60 ~ 120 up to more than 32.5%, improve 10 ~ 20% than prior art.
3, the enriching and purifying macroporous resin gardenia blue technology used in present invention process, can make the gardenia blue finished product look valency of manufacture up to 120.
4, this technique can operate (room temperature 20 ± 5 DEG C) at normal temperatures, and good stability, has broad application prospects.
Accompanying drawing explanation
Fig. 1 is amino acid screening Selection experiment result figure.
Embodiment
The present invention is further illustrated below by way of specific embodiment.But the detail of embodiment only for explaining the present invention, should not be construed as limited overall technical solution.Wherein, complex cellulase and glycine are all purchased from Huzhou Llilly Biotechnology Co., Ltd., and jasminoidin is purchased from Tian Zhirun bio tech ltd, Shaanxi.
Embodiment 1:
A) enzymolysis jasminoidin: jasminoidin 4kg, add complex cellulase (1,800,000 unit of activity) 4kg, anhydrous sodium acetate (analytical pure) 1.8kg, Glacial acetic acid (analytical pure) 980mL, purified water 100L, mixing, obtain jasminoidin enzymolysis feed liquid, stir, at reactor (Jiangsu Yang Yang chemical industry equipment Manufacturing Co., Ltd, 0.1m
3, 10,S34 II type) in stirring reaction 24h, 14000rmin under 15 DEG C of conditions
-1continuous centrifugal (sky, PVG this centrifugal machine company limited CQ145 high speed channel separator) 20min, lower sediment discards, and collects supernatant liquor;
B) develop the color: supernatant liquor adds glycine, and jasminoidin and glycine mol ratio are 1:1, in a kettle., stirring reaction 2.5h, 14000rmin at 20 DEG C
-1continuous centrifugal 20min, discards flocks, collects upper solution and obtains extracting solution;
C) enriching and purifying: with HPD417 resin column enriching and purifying, resin upper column quantity is 100L extracting solution (column volume is about 150L), use tap water and 70% ethanol elution respectively, collect water elution liquid and 4 times of column volume 70% ethanol eluates of 4 times of column volumes, respectively elutriant is concentrated, select single-action concentration tank that 4 times of column volume tap water washing lotions are concentrated into about 50kg, 4 times of column volume 70% alcohol washing lotions are concentrated into about 30kg, and thickening temperature is about 15 DEG C; Adopt spray dried form to carry out obtaining cream to concentrated solution, the sample introduction speed of adjustable spraying drying, make concentrated after water elution liquid temperature out be 85 ~ 90 DEG C, inlet temperature is 170 ~ 180 DEG C, Dry Sack valency 20 ~ 40 gardenia blue 6.5Kg; Ethanol eluate temperature out after concentrated is 75 ~ 80 DEG C, and inlet temperature is 170 ~ 180 DEG C, Dry Sack valency 60 ~ 120.1 gardenia blue 1.3Kg; Jasminoidin transformation efficiency is about 90%.
Embodiment 2:
A) enzymolysis jasminoidin: jasminoidin 4kg, add complex cellulase (1,600,000 unit of activity) 6kg, anhydrous sodium acetate (analytical pure) 1.5kg, Glacial acetic acid (analytical pure) 980mL, purified water 100L, mixing, obtain jasminoidin enzymolysis feed liquid, stir, at reactor (Jiangsu Yang Yang chemical industry equipment Manufacturing Co., Ltd, 0.1m
3, 10,S34 II type) in stirring reaction 36h, 10000rmin under 50 DEG C of conditions
-1continuous centrifugal (sky, PVG this centrifugal machine company limited CQ145 high speed channel separator) 20min, lower sediment discards, and collects supernatant liquor;
B) develop the color: supernatant liquor adds glycine, and jasminoidin and glycine mol ratio are 1:1.5, in a kettle., stirring reaction 2.5h, 14000rmin at 60 DEG C
-1continuous centrifugal 25min, discards flocks, collects upper solution and obtains extracting solution;
C) enriching and purifying: with HPD600 type macroporous adsorptive resins enriching and purifying, resin upper column quantity is 100L extracting solution (column volume is about 150L), use tap water and 50% ethanol elution respectively, collect water elution liquid and 4 times of column volume 60% ethanol eluates of 4 times of column volumes, respectively elutriant is concentrated, select single-action concentration tank that 4 times of column volume tap water washing lotions are concentrated into about 50kg, 4 times of column volume 60% ethanol washing lotions are concentrated into about 30kg, and thickening temperature is about 50 DEG C; Adopt spray dried form to carry out obtaining cream to concentrated solution, the sample introduction speed of adjustable spraying drying, make concentrated after water elution liquid temperature out be 85 ~ 90 DEG C, inlet temperature is 170 ~ 180 DEG C, Dry Sack valency 20 ~ 40 gardenia blue 6.4Kg; Ethanol eluate temperature out after concentrated is 75 ~ 80 DEG C, and inlet temperature is 170 ~ 180 DEG C, Dry Sack valency 60 ~ 120.1 gardenia blue 1.28Kg; Jasminoidin transformation efficiency is about 90%.
Embodiment 3:
A) enzymolysis jasminoidin: jasminoidin 4kg, add complex cellulase (2,000,000 unit of activity) 12kg, anhydrous sodium acetate (analytical pure) 1.2kg, Glacial acetic acid (analytical pure) 980mL, purified water 120L, mixing, obtain jasminoidin enzymolysis feed liquid, stir, at reactor (Jiangsu Yang Yang chemical industry equipment Manufacturing Co., Ltd, 0.1m
3, 10,S34 II type) in stirring reaction 12h, 18000rmin under 70 DEG C of conditions
-1continuous centrifugal (sky, PVG this centrifugal machine company limited CQ145 high speed channel separator) 20min, lower sediment discards, and collects supernatant liquor;
B) develop the color: supernatant liquor adds glycine, and jasminoidin and glycine mol ratio are 1.5:1, in a kettle., stirring reaction 3h, 18000rmin at 100 DEG C
-1continuous centrifugal 15min, discards flocks, collects upper solution and obtains extracting solution;
C) enriching and purifying: with HPD417 resin column enriching and purifying, resin upper column quantity is 100L extracting solution (column volume is about 150L), use tap water and 70% ethanol elution respectively, collect water elution liquid and 4 times of column volume 70% ethanol eluates of 4 times of column volumes, respectively elutriant is concentrated, select single-action concentration tank that 4 times of column volume tap water washing lotions are concentrated into about 50kg, 4 times of column volume 70% alcohol washing lotions are concentrated into about 30kg, and thickening temperature is about 70 DEG C; Adopt spray dried form to carry out obtaining cream to concentrated solution, the sample introduction speed of adjustable spraying drying, make concentrated after water elution liquid temperature out be 85 ~ 90 DEG C, inlet temperature is 170 ~ 180 DEG C, Dry Sack valency 20 ~ 40 gardenia blue 6.3Kg; Ethanol eluate temperature out after concentrated is 75 ~ 80 DEG C, and inlet temperature is 170 ~ 180 DEG C, Dry Sack valency 60 ~ 120.1 gardenia blue 1.26Kg; Jasminoidin transformation efficiency is about 90%.The quality determining method of gardenia blue:
1, HPLC method detects:
Select commercially available look valency be the gardenia blue sample (Shantou pleasant virtue company natural gardenia blue MD-GB30) of 40 as with reference to product, Liquid Detection self-control gardenia blue sample.
Chromatographic condition: PhenomenexBiosep-SEC-S2000(300mm × 7.80mm) gel column, moving phase is purified water, and elution time is 30min, and sample size is 10 μ L, and column temperature is 30 DEG C, and determined wavelength is 590nm, and flow velocity is 0.5mLmin
-1, type of elution is isocratic elution, and it is 20 μ gmL that commercial samples and self-control sample concentration are joined
-1.
Integrative approach: slope sensitivity 1, peak width 0.45, smallest peaks area 5, peak height 0, acromion is closed.
2, UV method detects:
Sample preparation:
1. commercially available gardenia blue sample preparation, takes 0.0195g sample, and constant volume, in the purified water of 100mL, keeps sample to be measured;
2. embodiment gardenia blue sample preparation, takes 0.0296g sample, and constant volume, in the purified water of 500mL, keeps sample to be measured.
Detection method: under 590nm absorbing wavelength, surveys commercially available and self-control sample absorbance.
Calculation formula:
a representative sample absorbancy at 590 nm, f representative sample dilution volume, m representative sample sampling quality, 100 is reduction factor.
Effective look valency calculation formula:
E1, E2, Ex representative color valency be greater than 40 respectively wash mutually respectively look valency, m1, m2, mx representative color valency be greater than 40 respectively wash mutually cream weight;
Effective yield calculates: k
effectively=k
1+ k
2+ k
x, the wash-out phase wash-out yield sum that look valency is greater than 40, k represents wash-out yield;
Detected result: in table 4.
Table 4
Claims (9)
1. a preparation method for gardenia blue, is characterized in that the method comprises the following steps:
A) enzymolysis jasminoidin: take water as solvent, adds jasminoidin, complex cellulase, anhydrous sodium acetate and Glacial acetic acid, stirring reaction 12 ~ 36 hours under 15 ~ 70 DEG C of conditions; Wherein, complex cellulase and jasminoidin mass ratio 0.5 ~ 1.5:0.5 ~ 1.5, anhydrous sodium acetate and jasminoidin mass ratio 0.3 ~ 0.5:0.6 ~ 1.2, Glacial acetic acid and jasminoidin mass ratio 0.1 ~ 0.4:0.8 ~ 1.2; By the solution centrifugal obtained after enzymolysis, discard precipitation, collect supernatant liquor;
B) develop the color: supernatant liquor is added glycine according to the ratio that jasminoidin and glycine mol ratio are 0.5 ~ 1.5:0.5 ~ 1.5 and develops the color; By the solution centrifugal obtained after colour developing, discard precipitation, collect upper solution, obtain extracting solution; Described color condition is develop the color 1 ~ 3 hour at 20 ~ 100 DEG C;
C) enriching and purifying: adopting macroporous adsorptive resins or hydrogen bonding adsorption resin post enriching and purifying, after washing 2 ~ 6 times of column volumes, is that 2 ~ 6 times of column volumes washed by 50% ~ 70% ethanol by concentration, collects water elution liquid and ethanol eluate respectively; Water elution liquid concentrates, spraying dry obtain look valency 20 ~ 40 gardenia blue, ethanol eluate concentrates, spraying dry obtains the gardenia blue of look valency more than 60.
2. the preparation method of gardenia blue according to claim 1, is characterized in that the enzyme adopted in the enzymolysis jasminoidin step described in the method is the complex cellulase of 160 ~ 2,000,000 unit of activity.
3. the preparation method of gardenia blue according to claim 1, is characterized in that the water consumption described in the method is every 1kg jasminoidin water 20 ~ 30L; Described centrifugal condition is 10000 ~ 18000rmin
-1centrifugal 15 ~ 25min under condition.
4. the preparation method of gardenia blue according to claim 1, is characterized in that the macroporous adsorbent resin that enriching and purifying step adopts is polarity or nonpolar macroporous adsorption resin.
5. the preparation method of gardenia blue according to claim 1, it is characterized in that the macroporous adsorbent resin that enriching and purifying step adopts is HPD600 type macroporous adsorbent resin, described hydrogen bonding adsorption resin is HPD417 type polymeric adsorbent.
6. the preparation method of gardenia blue according to claim 1, is characterized in that the elution speed adopted in enriching and purifying step is 1 ~ 3BV/h, and applied sample amount is that every 1.5ml column volume adds 0.5 ~ 1ml extracting solution.
7. the preparation method of gardenia blue according to claim 1, is characterized in that in enriching and purifying step, the concentrated temperature adopted is 15 ~ 70 DEG C.
8. the preparation method of gardenia blue according to claim 1, the condition that the water elution liquid spraying dry after to it is characterized in that described in the method concentrated adopts is: inlet temperature 170 ~ 180 DEG C, temperature out is 85 ~ 90 DEG C.
9. the preparation method of gardenia blue according to claim 1, the condition that the ethanol eluate spraying dry after to it is characterized in that described in the method concentrated adopts is: inlet temperature 170 ~ 180 DEG C, temperature out is 75 ~ 80 DEG C.
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| CN105017806B (en) * | 2015-08-31 | 2017-03-29 | 桂林茗兴生物科技有限公司 | A kind of method that gardenia blue pigment is prepared from Fructus Gardeniae |
| CN105861564A (en) * | 2016-06-27 | 2016-08-17 | 赣州华汉生物科技有限公司 | Method of purifying gardenia blue pigment resin |
| CN113402460B (en) * | 2021-05-10 | 2022-08-26 | 南京中医药大学 | Gardenia blue pigment with anti-mental disease function and preparation method and application thereof |
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| JPH07111896A (en) * | 1993-10-19 | 1995-05-02 | Taito Kk | Production of clear-colored blue pigment |
| CN102021202A (en) * | 2009-09-18 | 2011-04-20 | 中国科学院兰州化学物理研究所 | Method for preparing gardenia blue pigment |
| CN102021210A (en) * | 2009-09-18 | 2011-04-20 | 桂林莱茵生物科技股份有限公司 | Method for preparing gardenia blue pigment |
| CN102732050A (en) * | 2012-06-06 | 2012-10-17 | 浙江科技学院 | Method for preparing gardenia pigments from Gardenia jasminoides Ellis |
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- 2012-10-22 CN CN201210404931.9A patent/CN103773823B/en active Active
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07111896A (en) * | 1993-10-19 | 1995-05-02 | Taito Kk | Production of clear-colored blue pigment |
| CN102021202A (en) * | 2009-09-18 | 2011-04-20 | 中国科学院兰州化学物理研究所 | Method for preparing gardenia blue pigment |
| CN102021210A (en) * | 2009-09-18 | 2011-04-20 | 桂林莱茵生物科技股份有限公司 | Method for preparing gardenia blue pigment |
| CN102732050A (en) * | 2012-06-06 | 2012-10-17 | 浙江科技学院 | Method for preparing gardenia pigments from Gardenia jasminoides Ellis |
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