CN102827894B - Method for separating anthocyanin pigment and lactic acid from radish fermentation broth - Google Patents
Method for separating anthocyanin pigment and lactic acid from radish fermentation broth Download PDFInfo
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- C07C51/42—Separation; Purification; Stabilisation; Use of additives
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- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
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- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/56—Lactic acid
Abstract
The invention discloses a method for separating anthocyanin pigment and lactic acid from radish fermentation broth. According to the method, radishes are fermented by using lactobacillus plantarum, and then obtained fermentation broth is separated by using column chromatography so as to simultaneously extract anthocyanin pigment and lactic acid. The method provided by the invention has the advantages of high yield of anthocyanin pigment and lactic acid, a good separation effect, a simple process and suitability for industrial large-scale preparation.
Description
Technical field
The present invention relates to a kind of method of simultaneously extracting pigment and lactic acid, particularly from radish fermented liquid, extract the method for radish anthocyani pigment and lactic acid simultaneously.
Background technology
Radix Dauci Sativae (Radish) is a class for Cruciferae (Cruciferae) Rhaphanus (Raphanus L) radish, wide in variety, such as can be divided into the white heart of red skin, red skin red heart, shagreen turnip with red inside etc. from colour generation feature.Radix Dauci Sativae is rich in anthocyani pigment, is commonly called as radish red pigment ，Yi China and gets the Green Light as natural additive for foodstuff.Thereby the radish of being rich in haematochrome is widely used in tradition and modern fermentation industry as upper color substance.For artificial haematochrome, that radish red pigment has is safe and reliable, have no side effect, tone nature, have the advantages such as certain pharmacological action and nutritive effect.Except coloring matter, radish fermented liquid is being rich in and is being worth high lactic acid.It is a kind of multiduty fine chemicals, can be widely used in the industries such as food, pharmacy, environmental protection, agricultural.And its product main forms is acidic flavoring agent, seasonings, sanitas, tanning agents, plant-growth regulator, Biodegradable material and chiral drug etc.But at radish fermented liquid, after fermentation completes, be used as at present Industrial Wastewater Treatment, the not only wasting of resources, also causes environmental pollution.By literature search, there is no the pertinent literature patent report about pigment in radish fermented liquid and lactic acid comprehensive utilization aspect.Therefore, the core content of this patent is the higher value application of anthocyani pigment and lactic acid in the fermented abandoned liquid of radish.
About preparing the method for radish red pigment, mainly extract and synthesize.Chinese patent (CN1241599), carries out polygalacturonase processing to the vat liquor of Radix Dauci Sativae, then crosses post absorption and purification pigment, and then red liquid is collected in decolouring, through vacuum compression and the dry radish anthocyani pigment that obtains of spraying.Chinese patent (CN1510086), to fresh Radix Dauci Sativae lixiviate extraction, then uses resin absorption, last spray-dried concentrated, vacuum combination drying and remove peculiar smell and process and obtain radish red pigment powder.Chinese patent (CN101525498), by radish preheating squeezing, then carries out enzymolysis, then removes stink by the adsorptivity of chitosan, and concentrate drying obtains radish anthocyanogen pigment powder.Chinese patent (CN102093748A) be take turnip with red inside as raw material, through extraction, deproteinization, de-glucorphanin, ultrafiltration purification, concentrated, spraying is dry and make radish red pigment list aggressiveness and radish pycnogenols product.Chinese patent (CN101775233A), to turnip with red inside lixiviate, then micro-filtration, ultrafiltration, nanofiltration, makes highly purified radish anthocyani pigment.
Aspect the extraction preparation of lactic acid, Chinese patent (CN1603298) has been announced a kind of processing method of extracting lactic acid from fermented liquid, utilize gac and H-103 resin decolorization except sugar, twice cooling, primary crystallization obtains calcium lactate, resulting calcium lactate percent crystallization in massecuite can reach 80%, and purity reaches 97.2%.Chinese patent (CN101294169), employing has been announced a kind of new neutralizing agent and has been fermented, and starch material is sized mixing and inoculated the generation Sodium.alpha.-hydroxypropionate that ferments, and then carries out ultrafiltration, and film is analysed and is generated lactic acid and sodium hydroxide, and recovery lactic acid concentrates and obtains product.Chinese patent (CN1226601), announced a kind of method of extracting calcium lactate from distillery waste, add a certain amount of yellow water to mix fermentation waste water, then add liming sedimentation, filter, finally concentrated clean, the present invention is more than 81.0% to the extraction yield of calcium lactate, and easy and simple to handle, with low cost.
By above-mentioned, can be found out, mostly the extraction of radish anthocyani pigment is to take directly fresh Radix Dauci Sativae carried out to lixiviate extraction and obtain, and fermentation method is generally taked in the preparation of lactic acid, but adopt a set of separation, extraction flow process to come the technique of separated radish anthocyani pigment of while and lactic acid also unrealized.The up to ten million tons of output of the annual rouge radish of China, are wherein used for making pickles greatly, and China's pickle production technology and new technology development have good development.Yet the fermented liquid of pickles after by fermentation but abandons, and not only cause exhaust emission, and a large amount of pigments and the lactic acid that wherein contain all wasted.The present invention is intended to design a set of technical process, by lactic fermentation, and separated preparation radish anthocyani pigment and lactic acid.This technique is also applicable to China's pickles industry, i.e. anthocyani pigment and lactic acid in the fermented abandoned liquid of higher value application.
Summary of the invention
The object of the invention is to solve above-mentioned the deficiencies in the prior art, a kind of method of simultaneously extracting radish anthocyani pigment (hereinafter to be referred as pigment) and lactic acid from radish fermented liquid is provided, described method technique is simple, environmental protection.
For achieving the above object, the present invention is by the following technical solutions:
From radish, extract a method for radish anthocyani pigment and lactic acid simultaneously, comprise the steps:
(1) actication of culture spreads cultivation
The activation of milk-acid bacteria: by freezing plant lactobacillus Lactobacillus plantarum (bacterium number: CICC22696, be purchased from Institute of Microorganism, Academia Sinica) bacterial classification thaw after, choose 2～3 rings and be inoculated on 300mL potato substratum, at 30 ℃, cultivate 18～24h;
Spreading cultivation of milk-acid bacteria: by volume 5%, in the potato substratum that the potato substratum that is rich in above activated spawn is spread cultivation new, at 30 ℃, cultivate 18-24h and can be used for radish inoculation;
Described potato substratum making method: boil 500mL water, then take the potato block 120g that cleans and cut, boil and be incubated 30 minutes, then filter, add 12g white sugar again, filtrate is distributed into two Erlenmeyer flasks, each quantitatively arrives 300mL with deionized water; Then with the soft rubber ball that is surrounded by gauze, seal bottleneck, then wrap up with newspaper, and tie with bungee, put into Autoclave sterilizing about 2 hours.
(2) pre-treatment, fermentation
Radish is cleaned, peeled, blanching 3～7min enzyme that goes out in 95～100 ℃ of water then, preferably radish blanching enzyme time of going out is 3min, is cooled to rapidly 10～20 ℃ after heating; The Sterile Saline of radish skin after enzyme and 8g/100mL of going out packs in sterilized fermentor tank than (g/ml) according to 3: 6～7 mass/volume, the two ratio preferably 3: 7; Then, by volume 5%, the plant lactobacillus activating after spreading cultivation is inoculated in fermentor tank;
(3) fermentation stops
Ferment after 7～14 days, collect fermented liquid, and carry out high temperature (90-121 ℃) sterilising treatment;
(4) preliminary purification
By fermented liquid centrifugal (10000g, 4 ℃, 5～10min), collect supernatant liquor;
(5) radish anthocyani pigment and lactic acid is separated
Utilize polymer packing material (macroporous adsorbent resin or C18 resin) chromatography column, take adsorption method of separation to obtain the separated of pigment and lactic acid, use subsequently aqueous ethanolic solution (80-100%, v/v) desorb pigment, thus pigment and the lactic acid of supernatant liquor described in separated (4).Concrete steps are for to adjust to by supernatant liquor pH value (3.4～3.5) described in (4) supernatant liquor that pH < 3 obtains acidifying with hydrochloric acid (6mol/L); According to the supernatant liquor of acidifying, hydrochloric acid (v/v) deionized water containing 0.01%, hydrochloric acid (v/v) aqueous ethanolic solution containing 0.01%, carry out application of sample, collect respectively elutriant: ethanol is collected liquid (A) and is rich in coloring matter, other collection liquid (B) are rich in lactic acid material; Described macroporous adsorbent resin is that (specific surface area is 500m to XAD-7HP
2/ g; Mean pore size is 450
; Median size is 560 μ m; Dipole moment is 1.8; Coefficient of uniformity D90/D40 is 1.7); Described C18 resin is the anti-phase RP-18 filler of Lichrospher (aperture 100A, particle diameter 25-40 μ m); Described chromatography column size is
or 10mL with 2mL injection needles cylinder as the separated chromatography column of sample;
(6) radish anthocyani pigment and lactic acid is refining
Through above-mentioned resin isolation, obtain elutriant, ethanol is collected to liquid (A) centrifugal (10000g, 4 ℃, 5～10min), collect supernatant liquor, vacuum concentration to volume is less than 1/4th of original volume; Again this concentrated solution is carried out to centrifugal (10000g, 4 ℃, 5～10min), collect supernatant liquor, add ethanol or aqueous ethanolic solution, make final alcohol concn be greater than 80% (v/v), carry out alcohol precipitation, centrifugal (10000g, 4 ℃, 5～10min), evaporation concentration to volume and be less than 1/4th of original volume, 1～4 time so repeatedly, then to obtain garnet powder be described radish anthocyani pigment to evaporate to dryness; Other are collected to liquid (B) concentrate, centrifugal (10000g, 4 ℃, 5～10min), collect supernatant liquor, then with ethanol, carry out alcohol precipitation, centrifugal (10000g, 4 ℃, 5～10min), collect supernatant liquor, be concentrated into and remove all ethanol, obtain the lactic acid concentrated solution of purifying, by vacuum-drying or lyophilize, obtaining powder mass is described lactic acid.
In above-mentioned steps (5) when separated pigment and lactic acid, water-soluble very strong due to lactic acid, in theory with very weak with nonpolar extremely strong resin affinity, so the adsorptive capacity in resin is less; For lactic acid, pigment is nonpolar stronger, and resin is strong to the adsorptivity of pigment.Therefore, in water solution system, pigment can be attracted on resin, and lactic acid can not be attracted on resin.
Radish of the present invention is the radish that contains anthocyani pigment, heart radish as white in red skin, red skin turnip with red inside, shagreen turnip with red inside etc.
Advantage of the present invention is: according to the physico-chemical property of radish anthocyani pigment (hereinafter to be referred as pigment) and lactic acid, set up the separation preparation of pigment and lactic acid in Radix Dauci Sativae fermented liquid, the pigment that this technique makes and lactic acid yield are high, good separating effect and simple process.Utilize the Selective adsorption of resin to pigment and lactic acid, C18 resin material and macroporous adsorbent resin material reach separating effect: pigment yield is respectively 90.8% (C18 resin) and 94.0% (macroporous resin), finally take rotary evaporation that concentrated solution evaporate to dryness is made to pigment powder, it is garnet powder, yield is more than 84.5%, more than purity reaches 2.6% (w/w); Lactic acid yield is 89.9% (C18 resin) and 81.0% (macroporous resin), more than purity reaches 53.3% (w/w).
Accompanying drawing explanation
Fig. 1 is radish fermented liquid anthocyani pigment and lactic acid preparation technology flow process.
Embodiment
Below by embodiment, the present invention is specifically described, but technical solution of the present invention is not limited to following cited embodiment.
In following examples, big pore adsorption resin is that (specific surface area is 500m to XAD-7HP
2/ g, mean pore size is 450
, median size is 560 μ m, dipole moment is 1.8, coefficient of uniformity D90/D40 is 1.7); C18 resin used is the anti-phase RP-18 filler of Lichrospher (aperture 100A, particle diameter 25-40 μ m); Chromatography column size used is
In following examples, potato substratum making method used is: boil 500mL water, then take the potato block 120g that cleans and cut, boil and be incubated 30 minutes, then filter, add 12g white sugar again, filtrate is distributed into two Erlenmeyer flasks, each quantitatively arrives 300mL with deionized water; Then with the soft rubber ball that is surrounded by gauze, seal bottleneck, then wrap up with newspaper, and tie with bungee, put into Autoclave sterilizing about 2 hours.
In following examples, preparation process as shown in Figure 1.
Embodiment 1
Freezing plant lactobacillus Lactobacillus plantarum (bacterium number: CICC 22696) is inoculated into the activation of thawing on 300mL potato substratum, cultivates 18～24h.By volume 5%, in the potato substratum that the potato substratum that is rich in above activated spawn is spread cultivation new, at 30 ℃, cultivate 18～24h and can be used for radish inoculation; Red skin white turnip is cleaned, peel, 100 ℃ of blanching 7min then, go out radish skin after enzyme and the Sterile Saline of 8g/100mL pack in sterilized fermentor tank according to the mass/volume ratio of 300g: 600mL, by volume 5%, the plant lactobacillus activating after spreading cultivation is inoculated in fermentor tank; Ferment 7 days, collect fermented liquid, and carry out high temperature (90～121 ℃) sterilising treatment; By fermented liquid, centrifugal (10000g,, 10min), collects supernatant liquor (be the front anthocyanogen of purifying and lactic mixt, table 1) by 4 ℃; With 36g macroporous adsorbent resin, fill dress post, then according to the fermented liquid of 50mL acidifying, 50mL carries out application of sample and elutriant containing deionized water, the 100mL of 0.01% hydrochloric acid (v/v) containing the order of the dehydrated alcohol of 0.01% hydrochloric acid (v/v), collect respectively elutant, ethanol is collected liquid (A) and is rich in coloring matter, and other collection liquid (B) are rich in lactic acid material; 100mL ethanol is collected to the centrifugal (10000g of liquid (A), 4 ℃, 10min), collect supernatant liquor, carry out evaporation concentration to 10～20mL, then centrifugal (10000g, 4 ℃, 10min), take out supernatant liquor, add 40～80mL dehydrated alcohol to carry out alcohol precipitation, then centrifugal (10000g, 4 ℃, 10min), evaporation concentration to 10～20mL, 3～4 times to without obvious sediment so repeatedly, then evaporate to dryness remaining liq, obtain garnet pigment powder (for the anthocyanogen after purification process, table 1); 100mL fermented liquid and deionized water collection liquid (B) is concentrated, centrifugal (10000g, 4 ℃, 10min), collect supernatant liquor, then with ethanol, carry out alcohol precipitation, centrifugal (10000g, 4 ℃, 10min), collect supernatant liquor, carry out again lyophilize, obtain being rich in the solid matter (for the lactic acid sample after purification process, table 1) of lactic acid.
Embodiment 2
Radish blanching enzyme time of going out is 3min, and other steps are with embodiment 1.
Embodiment 3
The mass/volume ratio of the Sterile Saline of radish skin and 8g/100mL is 300g: 700mL, and other steps are with embodiment 1.
Embodiment 4
Fermented liquid is 5min in the centrifugal treating time of purifying, treating process, and other steps are with embodiment 1.
Embodiment 5
Utilize the separation of C18 resin absorption to prepare in the process of fermented liquid pigment and lactic acid, according to the fermented liquid of 100mL acidifying, 100mL carries out application of sample and elutriant containing deionized water and the 200mL of 0.01% hydrochloric acid (v/v) containing the order of the dehydrated alcohol of 0.01% hydrochloric acid (v/v), and other steps are with embodiment 1.
Embodiment 6
Change the amount of the deionized water in embodiment 5 into 150mL, other steps are with embodiment 1.
Embodiment 7
Preceding step is with embodiment 1, in separation and purification process, macroporous resin loading level changes 3.6g into, with 10mL injection needle tube as chromatography column, then the amount of the dehydrated alcohol of 5mL acidifying fermentation liquid, 5mL acidifying deionized water, 10mL acidifying changes according to the corresponding proportion in embodiment 1, collect ethanol collection liquid (A) and acidifying fermentation liquid, acidifying deionized water collection liquid (B), measure respectively radish anthocyani pigment and the lactic acid content of collecting in liquid A, B.
Embodiment 8
With C18 (0.5g) resin alternate embodiment 7 macroporous adsorbent resins, with 2mL injection needle tube as chromatography column, then according to the order of the dehydrated alcohol of the deionized water of the fermented liquid of 1mL acidifying, 1mL acidifying, 2mL ethyl acetate, 2mL acidifying, carry out application of sample and elutriant, other are with embodiment 7.
Embodiment 9
200mL radish anthocyani pigment elutant, as after alcohol precipitation in embodiment 1 processes, is concentrated into 10～20mL, and other steps are with embodiment 1, pre-freeze 18～24h at-80 ℃ then, then carry out lyophilize.
Performance test experiment:
1, the mensuration of radish anthocyanogen pigment content
Fermented liquid is used respectively to the potassium chloride buffer solution of pH 1.0,0.025M and 10 times of the sodium acetate buffer solution dilutions of pH 4.5,0.4M (every 300 μ L are diluted to 3mL), under UV-1800 ultraviolet spectrophotometer, detect respectively 520nm and 700nm absorbance value A.Then be calculated as follows the content of radish anthocyani pigment:
In formula:
DF---extension rate;
MW---molecular weight; The anthocyanogen containing in radish is mainly pelargonidin, so the MW molecular weight that is pelargonidin, 433.2;
The molar absorptivity of ε---pelargonidin, its value is 31600;
L---light path, 1cm; Pigment is radish anthocyani pigment.
The potassium chloride buffer solution of secure ph 1.0,0.025M: accurately take 1.86g Repone K, add the distilled water of 980mL, measure pH value, regulate pH value to 1.0 with concentrated hydrochloric acid, use distilled water constant volume to 1.0L.
The sodium acetate buffer solution of secure ph 4.5,0.4M: accurately take the Sodium acetate trihydrate of 54.43g, be dissolved in the distilled water of 960mL, regulate pH value to 4.5 with concentrated hydrochloric acid, use distilled water constant volume to 1.0L.
2, the mensuration of lactic acid content
With Agilent 1260 type high performance liquid chromatographs, survey lactic acid content.Liquid-phase condition is: Aligen ZORBAX SB-Aq pillar (150 * 4.6mm, 5 μ m), detection wavelength is 210nm, flow velocity is 0.8mL/min, sample size 30ul, mobile phase composition is A phase (formic acid: methyl alcohol: water=0.1: 3: 96.9, v/v/v), with B phase (acetonitrile), condition of gradient elution was: 0～5min:100%A phase; 5～20min:100%-50%A phase; 20～20.01min:50%-100%A phase; 20.01～28min 100%A phase.With HPLC, with Pfansteihl mark product (Sigma, quality purity 98%), make lactic acid typical curve.
Test result
In embodiment 1～9, by having changed the consumption of some conditions, but resulting result is more or less the same.The change of blanching time in embodiment 2, the change of inoculative proportion in embodiment 3, the pigment in the Radix Dauci Sativae fermented liquid material finally obtaining and the content of lactic acid do not have too notable difference.The change of centrifugation time in embodiment 4, can cause centrifugal effect to be not so good as the good of 10min.In embodiment 5 and 6, the corresponding change of application of sample amount does not have influence on the separating effect of pigment and lactic acid, and the rate of recovery is also comparatively stable.Embodiment 7 explanation quantitative changes are little, and circulation ratio is better.Example 8 is selected C18 resin, the similar results obtaining.Example 9 increases radish fermentation liquor treatment amount, and radish anthocyani pigment elutant is carried out to lyophilize, and in pigment powder, anthocyanogen purity is 3.36% (w/w).Take embodiment 1 as example, by rotary evaporation, can access bolarious powder, lactic acid after alcohol precipitation, dried data are as shown in table 1:
The purity of anthocyanogen and lactic acid and the rate of recovery in table 1 embodiment 1
Anthocyanogen (or lactic acid) total content before anthocyanogen after the rate of recovery=purifying (or lactic acid) total content * 100%/purifying
Purification process comprises column chromatography for separation, alcohol precipitation.
Of the present invention a kind of from Radix Dauci Sativae fermented liquid the technique of separated pigment and lactic acid by concrete example, be described, those skilled in the art can use for reference content of the present invention, the links such as appropriate change raw material, processing condition realize corresponding other object, its relevant change does not all depart from content of the present invention, within all similar replacements and change will become apparent to those skilled in the art that and be all deemed to be included in scope of the present invention.
Claims (4)
1. from radish, extract a method for radish anthocyani pigment and lactic acid simultaneously, comprise the steps:
(1) actication of culture spreads cultivation
The activation of milk-acid bacteria: will buy in Institute of Microorganism, Academia Sinica, bacterium number is, after the bacterial classification of the freezing plant lactobacillus of CICC22696 thaws, to be inoculated on 300mL potato substratum, cultivates at 30 ℃ the potato substratum that 18～24h must be rich in activated spawn;
Spreading cultivation of milk-acid bacteria: in the potato substratum that the described potato substratum that is rich in activated spawn is spread cultivation new, cultivate 18-24h at 30 ℃ and can be used for radish inoculation;
(2) pre-treatment, fermentation
Radish is cleaned, peeled, and then the blanching 3min enzyme that goes out in 95～100 ℃ of water, is cooled to rapidly 10～20 ℃ after heating; Go out radish skin after enzyme and the Sterile Saline of 8g/100mL packs in sterilized fermentor tank according to the mass volume ratio of 3g ︰ 7ml; Then, by volume 5%, the plant lactobacillus after spreading cultivation is inoculated in fermentor tank;
(3) fermentation stops
Ferment after 7～14 days, collect fermented liquid, and carry out high-temperature sterilization processing;
(4) preliminary purification
Fermented liquid after sterilizing is centrifugal, collect supernatant liquor;
(5) radish anthocyani pigment and lactic acid is separated
With the hydrochloric acid of 6mol/L, supernatant liquor pH value in described (4) is adjusted to the supernatant liquor that pH<3 obtains acidifying, then utilize the separated pigment of polymer packing material chromatography post and lactic acid, application of sample is sequentially followed successively by: the supernatant liquor of described acidifying, containing 0.01%(v/v) deionized water of hydrochloric acid, containing 0.01%(v/v) aqueous ethanolic solution of hydrochloric acid, collect respectively elutriant: ethanol is collected liquid (A) and is rich in coloring matter, other collection liquid (B) is rich in lactic acid material, and the concentration of described aqueous ethanolic solution is 80～100%(v/v);
(6) radish anthocyani pigment and lactic acid is refining
Described ethanol is collected to liquid (A) centrifugal, collect supernatant liquor, vacuum concentration to volume is less than 1/4th of original volume, carry out again centrifugally, collect supernatant liquor, add ethanol or aqueous ethanolic solution, make final alcohol concn be greater than 80%(v/v), carry out alcohol precipitation, centrifugal, evaporation concentration to volume is less than 1/4th of original volume, 1～4 time so repeatedly, then to obtain garnet powder be described radish anthocyani pigment to evaporate to dryness; By described other collect liquid (B) concentrate, centrifugal, collect supernatant liquor, then with ethanol, carry out alcohol precipitation, centrifugal, collect supernatant liquor, then be concentrated into and remove all ethanol, obtain the lactic acid concentrated solution of purifying, by vacuum-drying or lyophilize, obtaining powder mass is described lactic acid;
Described in described (1), on 300mL potato substratum, inoculum size is 2～3 rings, described in be rich in the potato substratum of activated spawn and the volume ratio of described new potato substratum is 5%; Described potato substratum making method is: boil 500mL water, then take the potato block 120g that cleans and cut, boil and be incubated 30 minutes, then filter, add 12g white sugar again, filtrate is distributed into two Erlenmeyer flasks, each quantitatively arrives 300mL with deionized water; Then with the soft rubber ball that is surrounded by gauze, seal bottleneck, then wrap up with newspaper, and tie with bungee, put into Autoclave sterilizing 2 hours;
Wherein, polymer packing material described in described (5) is macroporous adsorbent resin or C18 resin.
2. the method for claim 1, wherein described centrifugal condition is: 10000g, 4 ℃, 5～10min.
3. the method for claim 1, wherein described macroporous adsorbent resin is XAD-7HP.
4. the method for claim 1, wherein described C18 resin is the anti-phase RP-18 filler of Lichrospher.
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| JP2002017394A (en) * | 2000-07-03 | 2002-01-22 | Akio Tsukui | Method for extracting anthocyanin pigment by lactic acid fermentation |
| CN1510086A (en) * | 2002-12-25 | 2004-07-07 | 赵厚坤 | Preparation of radish red pigment |
| CN101760497A (en) * | 2009-12-07 | 2010-06-30 | 耿兆翔 | Microorganisms or/and biological enzyme extracting method for plant anthocyanin |
| CN101776659A (en) * | 2010-02-03 | 2010-07-14 | 北京市理化分析测试中心 | Method for detecting anthocyanin in red radish through high-performance liquid chromatography |
| CN101775233A (en) * | 2009-09-08 | 2010-07-14 | 新疆医科大学 | Membrane separation integrated technology-based preparation method for producing deodorized red radish pigment |
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|---|---|---|---|---|
| JP2002017394A (en) * | 2000-07-03 | 2002-01-22 | Akio Tsukui | Method for extracting anthocyanin pigment by lactic acid fermentation |
| CN1510086A (en) * | 2002-12-25 | 2004-07-07 | 赵厚坤 | Preparation of radish red pigment |
| CN101775233A (en) * | 2009-09-08 | 2010-07-14 | 新疆医科大学 | Membrane separation integrated technology-based preparation method for producing deodorized red radish pigment |
| CN101760497A (en) * | 2009-12-07 | 2010-06-30 | 耿兆翔 | Microorganisms or/and biological enzyme extracting method for plant anthocyanin |
| CN101776659A (en) * | 2010-02-03 | 2010-07-14 | 北京市理化分析测试中心 | Method for detecting anthocyanin in red radish through high-performance liquid chromatography |
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