CN103773823A - Preparation method of gardenia blue - Google Patents
Preparation method of gardenia blue Download PDFInfo
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- CN103773823A CN103773823A CN201210404931.9A CN201210404931A CN103773823A CN 103773823 A CN103773823 A CN 103773823A CN 201210404931 A CN201210404931 A CN 201210404931A CN 103773823 A CN103773823 A CN 103773823A
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Abstract
The invention discloses a preparation method of gardenia blue. Through geniposide enzymolysis, developing, enrichment purification, condensation and drying, gardenia blue having a color value of 20-40 and gardenia blue having a color value more than 60 are prepared. The preparation method can be operated easily and has good stability, a high high-value gardenia blue yield and a wide application prospect.
Description
Technical field
The invention belongs to natural pigment field, be specifically related to a kind of preparation method of gardenia blue.
Background technology
In recent years, natural pigment has obtained very fast development.But most of natural pigment of producing is all take red, yellow two class colors as main.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 (Gardenia blue), by the colour generation glycoside body of warm water extraction gained and the mixture (amino acids) of protein decomposition product for madder wort cape jasmine fruit, add beta-glucosidase, through fermentation, deactivation, filtration, evaporation, refining, dry forming.Be blue.Have data in literature to show, the consumption of Japan's gardenia blue in 2006 is 100 tons of left and right, the population that the China of comparing is a large amount of, and along with the continuous lifting of standard of living, the actual demand amount of China should be far above 100 tons/year.Main product is that look valency is 40 gardenia blue product on the market at present, and 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, in the Chinese invention patent that is 200710045237.1 at application number, disclose with beta-glucoside enzyme catalysis jasminoidin reaction and obtained genipin, and genipin and amino acid have been reacted to obtain to the technology of natural gardenia blue pigment; In the Chinese invention patent that is 200910114410.8 at application number, disclose with cape jasmine fruit powder, through extracting, filter, the process such as concentrated, upper macroporous resin column, wash-out, the separation of alumina adsorption post, cellulase hydrolysis catalysis, amino acid hydrolysis, obtain the technology of gardenia blue pigment; Disclose waste liquid after extracting take Gardenia Yellow as raw material at paper " two-step approach is produced the researchs of gardenia blue pigment processing condition ", made the technology of gardenia blue pigment by the two-step approach that beta-glucoside enzymic fermentation and enzymatic reaction are 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 is higher than 100 the report that rarely has.
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 look valency and low look valency simultaneously.The inventive method and raw material sources are extensive, and cost is low, and the product preparing has been taken into account the demand of high-end market and low-end market.
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, add jasminoidin, complex cellulase, anhydrous sodium acetate and Glacial acetic acid, stirring reaction 12～36 hours under 15～70 ℃ 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 obtaining after enzymolysis, discard precipitation, collect supernatant liquor;
B) colour developing: the ratio that is 0.5～1.5:0.5～1.5 according to jasminoidin and glycine mol ratio by supernatant liquor adds glycine colour developing; By the solution centrifugal obtaining 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, wash after 2～6 times of column volumes, is that 50%～70% ethanol is washed 2～6 times of column volumes by concentration, collects respectively water elution liquid and ethanol eluate; Water elution liquid is concentrated, the dry look valency that obtains of spraying is at 20～40 gardenia blue, and ethanol eluate is concentrated, the dry gardenia blue of look valency more than 60 that obtain of spraying.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 adopting 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 for to develop the color 1～3 hour at 20～100 ℃.The macroporous adsorbent resin that enriching and purifying adopts is polarity or nonpolar macroporous adsorption resin, and the macroporous adsorbent resin preferably adopting is HPD600 type macroporous adsorbent resin, and hydrogen bonding adsorption resin is HPD417 type polymeric adsorbent.The elution speed adopting 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 adopting is 15～70 ℃.
The dry condition adopting of spraying described in the inventive method is: inlet temperature is 170～180 ℃, and the water elution liquid temperature out after concentrating is 85～90 ℃, and the ethanol eluate temperature out after concentrating is 75～80 ℃.
Below adopt part experimental example 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, result demonstration, and under 590nm absorbing wavelength, the absorbancy of reaction later stage solution is higher, and gardenia blue component content is larger.Can find out clearly that by Fig. 1 colour developing is after 8 hours, use glycine colour developing to have notable difference with using L-glutamic acid colour developing, because the growing amount difference of gardenia blue, this is just related to the height of yield, so this process selection glycine can obviously improve must measuring of gardenia blue as developer, sample-pretreating method is identical, and yield increased value can be used absorbance ratio to ask for relatively.Develop the color after 8 hours, with respect to L-glutamic acid, select glycine, absorbancy has improved approximately 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, the HPD100 macroporous adsorptive resins of choosing, HPD400A macroporous adsorptive resins, ADS-17 macroporous adsorptive resins, HPD600 macroporous adsorptive resins, HPD417 hydrogen bonding adsorption resin post are further carried out to screening experiment according to the operation steps of embodiment 1, result shows that gardenia blue effective yield (being the rate of extract, the calculating according to look valency more than 40) and the look valency of HPD600 macroporous adsorptive resins, the acquisition of HPD417 hydrogen bonding adsorption resin post is higher.The results are shown in Table 1.
Table 1
Three, the drying process screening experiment of gardenia blue washing lotion after mistake resin.
Under the identical condition of stock liquid, select 100 ℃ of drying with water baths, concentrate+drying under reduced pressure of rotary evaporation, concentrate+vacuum freezedrying of rotary evaporation, spraying to be dried, result shows: spray-dired effect 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 is prepared look valency more than 60, and the yield of the gardenia blue that particularly color range is 60～120, up to more than 32.5%, has improved 10～20% than prior art.
3, the enriching and purifying macroporous resin gardenia blue technology using in technique of the present invention, can make the gardenia blue finished product look valency of manufacture up to 120.
4, this technique can operate (20 ± 5 ℃ of room temperatures) at normal temperatures, and good stability, has broad application prospects.
Accompanying drawing explanation
Fig. 1 is that test-results figure is selected in amino acid screening.
Embodiment
Further illustrate the present invention by specific embodiment below.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 be all purchased from Huzhou Li Lai Bioisystech 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, mixes, obtain jasminoidin enzymolysis feed liquid, stir, at reactor (Jiangsu Yang Yang chemical industry equipment Manufacturing Co., Ltd, 0.1m
3, 10S34 II type) in stirring reaction 24h under 15 ℃ of conditions, 14000rmin
-1continuous centrifugal (the PVG CQ145 of Tian Ben centrifugal machine company limited high speed channel separator) 20min, lower sediment discards, and collects supernatant liquor;
B) colour developing: supernatant liquor adds glycine, and jasminoidin and glycine mol ratio are 1:1, in reactor, stirring reaction 2.5h at 20 ℃, 14000rmin
-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 respectively tap water and 70% ethanol elution, collect water elution liquid and 4 times of column volume 70% ethanol eluates of 4 times of column volumes, respectively that elutriant is concentrated, select single-action concentration tank that 4 times of column volume tap water washing lotions are concentrated into 50kg left and right, 4 times of column volume 70% alcohol washing lotions are concentrated into 30kg left and right, and thickening temperature is 15 ℃ of left and right; Adopt spraying drying mode to carry out obtaining cream to concentrated solution, the sample introduction speed that adjustable spraying is dry, making the water elution liquid temperature out after concentrated is 85～90 ℃, and inlet temperature is 170～180 ℃, and Dry Sack valency is at 20～40 gardenia blue 6.5Kg; Ethanol eluate temperature out after concentrated is 75～80 ℃, and inlet temperature is 170～180 ℃, and Dry Sack valency is at 60～120.1 gardenia blue 1.3Kg; Jasminoidin transformation efficiency is in 90% left and right.
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, mixes, obtain jasminoidin enzymolysis feed liquid, stir, at reactor (Jiangsu Yang Yang chemical industry equipment Manufacturing Co., Ltd, 0.1m
3, 10S34 II type) in stirring reaction 36h under 50 ℃ of conditions, 10000rmin
-1continuous centrifugal (the PVG CQ145 of Tian Ben centrifugal machine company limited high speed channel separator) 20min, lower sediment discards, and collects supernatant liquor;
B) colour developing: supernatant liquor adds glycine, and jasminoidin and glycine mol ratio are 1:1.5, in reactor, stirring reaction 2.5h at 60 ℃, 14000rmin
-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 respectively tap water and 50% ethanol elution, collect water elution liquid and 4 times of column volume 60% ethanol eluates of 4 times of column volumes, respectively that elutriant is concentrated, select single-action concentration tank that 4 times of column volume tap water washing lotions are concentrated into 50kg left and right, 4 times of column volume 60% ethanol washing lotions are concentrated into 30kg left and right, and thickening temperature is 50 ℃ of left and right; Adopt spraying drying mode to carry out obtaining cream to concentrated solution, the sample introduction speed that adjustable spraying is dry, making the water elution liquid temperature out after concentrated is 85～90 ℃, and inlet temperature is 170～180 ℃, and Dry Sack valency is at 20～40 gardenia blue 6.4Kg; Ethanol eluate temperature out after concentrated is 75～80 ℃, and inlet temperature is 170～180 ℃, and Dry Sack valency is at 60～120.1 gardenia blue 1.28Kg; Jasminoidin transformation efficiency is in 90% left and right.
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, mixes, obtain jasminoidin enzymolysis feed liquid, stir, at reactor (Jiangsu Yang Yang chemical industry equipment Manufacturing Co., Ltd, 0.1m
3, 10S34 II type) in stirring reaction 12h under 70 ℃ of conditions, 18000rmin
-1continuous centrifugal (the PVG CQ145 of Tian Ben centrifugal machine company limited high speed channel separator) 20min, lower sediment discards, and collects supernatant liquor;
B) colour developing: supernatant liquor adds glycine, and jasminoidin and glycine mol ratio are 1.5:1, in reactor, stirring reaction 3h at 100 ℃, 18000rmin
-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 respectively tap water and 70% ethanol elution, collect water elution liquid and 4 times of column volume 70% ethanol eluates of 4 times of column volumes, respectively that elutriant is concentrated, select single-action concentration tank that 4 times of column volume tap water washing lotions are concentrated into 50kg left and right, 4 times of column volume 70% alcohol washing lotions are concentrated into 30kg left and right, and thickening temperature is 70 ℃ of left and right; Adopt spraying drying mode to carry out obtaining cream to concentrated solution, the sample introduction speed that adjustable spraying is dry, making the water elution liquid temperature out after concentrated is 85～90 ℃, and inlet temperature is 170～180 ℃, and Dry Sack valency is at 20～40 gardenia blue 6.3Kg; Ethanol eluate temperature out after concentrated is 75～80 ℃, and inlet temperature is 170～180 ℃, and Dry Sack valency is at 60～120.1 gardenia blue 1.26Kg; Jasminoidin transformation efficiency is in 90% left and right.The quality determining method of gardenia blue:
1, HPLC method detects:
Select the gardenia blue sample that commercially available look valency is 40 (the blue MD-GB30 of Shantou pleasant virtue company natural gardenia) conduct with reference to product, Liquid Detection self-control gardenia blue sample.
Chromatographic condition: Phenomenex Biosep-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 ℃, and detection wavelength is 590nm, and flow velocity is 0.5mLmin
-1, type of elution is isocratic elution, it is 20 μ gmL that commercially available sample 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, survey commercially available and self-control sample absorbancy.
Calculation formula:
the absorbancy of A representative sample under 590nm, f representative sample dilution volume, m representative sample sampling quality, 100 is reduction factor.
Effectively look valency calculation formula:
E1, E2, the look valency respectively of respectively washing mutually that Ex representative color valency is greater than 40, m1, m2, mx representative color valency be greater than 40 respectively wash mutually to obtain 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 (10)
1. a preparation method for gardenia blue, is characterized in that the method comprises the following steps:
A) enzymolysis jasminoidin: take water as solvent, add jasminoidin, complex cellulase, anhydrous sodium acetate and Glacial acetic acid, stirring reaction 12～36 hours under 15～70 ℃ 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 obtaining after enzymolysis, discard precipitation, collect supernatant liquor;
B) colour developing: the ratio that is 0.5～1.5:0.5～1.5 according to jasminoidin and glycine mol ratio by supernatant liquor adds glycine colour developing; By the solution centrifugal obtaining 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, wash after 2～6 times of column volumes, is that 50%～70% ethanol is washed 2～6 times of column volumes by concentration, collects respectively water elution liquid and ethanol eluate; Water elution liquid is concentrated, the dry look valency that obtains of spraying is at 20～40 gardenia blue, and ethanol eluate is concentrated, the dry gardenia blue of look valency more than 60 that obtain of spraying.
2. the preparation method of gardenia blue according to claim 1, is characterized in that the enzyme adopting 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 color condition described in development step for to develop the color 1～3 hour at 20～100 ℃.
5. 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.
6. 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 HPD600 type macroporous adsorbent resin, and described hydrogen bonding adsorption resin is HPD417 type polymeric adsorbent.
7. the preparation method of gardenia blue according to claim 1, is characterized in that the elution speed adopting 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.
8. the preparation method of gardenia blue according to claim 1, is characterized in that in enriching and purifying step, the concentrated temperature adopting is 15～70 ℃.
9. the preparation method of gardenia blue according to claim 1, is characterized in that the dry condition adopting of water elution liquid spraying after concentrated described in the method is: 170～180 ℃ of inlet temperatures, temperature out is 85～90 ℃.
10. the preparation method of gardenia blue according to claim 1, is characterized in that the dry condition adopting of ethanol eluate spraying after concentrated described in the method is: 170～180 ℃ of inlet temperatures, temperature out is 75～80 ℃.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105017806A (en) * | 2015-08-31 | 2015-11-04 | 桂林茗兴生物科技有限公司 | Method for preparing gardenia jasminoides ellis blue pigment from gardenia jasminoides ellis |
| 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 |
| CN113402460A (en) * | 2021-05-10 | 2021-09-17 | 南京中医药大学 | Gardenia blue pigment with mental disease resistance and preparation method and application thereof |
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Cited By (4)
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| CN105017806A (en) * | 2015-08-31 | 2015-11-04 | 桂林茗兴生物科技有限公司 | Method for preparing gardenia jasminoides ellis blue pigment from gardenia jasminoides ellis |
| 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 |
| CN113402460A (en) * | 2021-05-10 | 2021-09-17 | 南京中医药大学 | Gardenia blue pigment with mental disease resistance and preparation method and application thereof |
| 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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