CN102746266B - Method for preparing high-purity cornflower pigment - Google Patents
Method for preparing high-purity cornflower pigment Download PDFInfo
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- CN102746266B CN102746266B CN201210242158.0A CN201210242158A CN102746266B CN 102746266 B CN102746266 B CN 102746266B CN 201210242158 A CN201210242158 A CN 201210242158A CN 102746266 B CN102746266 B CN 102746266B
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Abstract
The invention relates to a method for preparing high-purity cornflower pigment. The method comprises the operation steps as follows: 1, extracting procyanidins, and preparing procyanidins solution using fresh tea tree root; 2, purifying the procyanidins solution, and preparing the procyanidins crude solution; 3, preparing the cornflower pigment methanol solution from the procyanidins crude solution; 4, purifying the cornflower pigment to obtain the cornflower pigment solution; and 5, carrying out rotary evaporation and other treatments on the cornflower pigment solution to obtain the cornflower pigment powder with the purity of above 98%, wherein the cornflower pigment powder is carmine, is difficult to dissolve in water, and is freely soluble in methanol. The method is simple to operate, no large expensive experimental apparatus is needed, and the adopted organic solvent is low in price and can be recovered, so the cost is low; and the method can effectively remove sugar, protein, other types of anthocyanidins and flavonoids compounds of the extract, and is suitable for obtaining the high-purity cornflower product.
Description
Technical field
The invention belongs to natural product chemistry field, the present invention relates to a kind of preparation method of high purity cyanidin.
Background technology
Cyanidin is a kind of in anthocyan compound, under natural condition, the cyanidin monomer of free state is seldom shown in, main its glucosides form of existence form, is often connected to form anthocyanidin glucosides with one or more glucose (glucose), rhamnosyl (rhamnose), semi-lactosi (galactose), wood sugar (xylose), pectinose (arabinose) etc. by glycosidic link.The cyanidin of glucosides is extensively present in flower, fruit, stem, the leaf of flowering plant (angiosperm), in the enchylema of root organ, be distributed in 27Ge section, in the plant of 72 genus, wherein especially abundant with Pericarpium Vitis viniferae, sorosis, blackberry, blueberry, blueberry, cherry, hawthorn, raspberry, purple cabbage equal size.
Cyanidin is the same with other kind anthocyanidin, is not only a kind of safe natural edible red pigment, and has antibacterial, anti-ageing, to reduce cancer and the cardiovascular disorder important physiological functions such as sickness rate.The Main Function of anthocyanidin is to remove free radical, and many and old relevant disease is as heart trouble, cancer, wrinkle, disease of eye, handkerchief gold disease and Alzheimer degenerative brain disorder etc., all relevant with the oxygenizement being caused by free radical.
The analytical procedure research, the physiologically active of anthocyanidin that at present both at home and abroad mainly concentrate on the evaluation of anthocyanidin resource distribution and the foundation of resources bank, anthocyanidin about the research of anthocyanidin are, 6 aspects such as the application of the structural stability of the high efficiency extraction of natural anthocyanidin glucosides and isolation technique research, anthocyanidin and molecular degradation Mechanism Study, anthocyanidin and product developments.The cyanidin monomer of commercially available 95% purity is 842.4 yuan/milligram of Renminbi (http://www.sigmaaldrich.com/china-mainland.html) now, there is no pertinent literature report prepared by cyanidin monomer.Utilize the cyanidin monomer of low-cost Tea root high-purity to there are very large market outlook.
Summary of the invention
The invention provides a kind of high purity cyanidin monomer technology of preparing, to prepare highly purified cyanidin.
The concrete operation step of preparation high purity cyanidin is as follows:
1.1, the extraction of pycnogenols
The Tea root that 20g is fresh is put into refiner, add 200 milliliters of extracting solutions, 1000 revs/min of rotating speeds, homogenate 5 minutes, collects homogenate; Under 4 ℃ of temperature, 5000 revs/min of conditions of rotating speed, centrifugation 10 minutes, collects supernatant liquor, obtains pycnogenols solution; Described extracting solution adds 29.5 ml waters and 0.5 milliliter of acetic acid to make by 70 milliliters of acetone;
1.2, the purifying of pycnogenols
Pycnogenols solution and pure water by volume 1:10 mix, and through AB-8 macroporous resin adsorption, by the methanol-eluted fractions of 1 liter of concentration 20%, then use 100 milliliters of pure methanol-eluted fractions, collect meoh eluate, obtain pycnogenols crude product solution;
1.3, cyanidin methanol solution produces
Pycnogenols crude product solution and pyrolysis solution by volume 1:10 mix, and 95 ℃ of pyrolysis of temperature 1 hour, obtain cyanidin pyrolysis solution; Utilize Rotary Evaporators, temperature 70 C water-bath evaporation 1 hour, removes pyrolysis solution, and 10 milliliters of pure dissolve with methanol for residual dry powder, obtain cyanidin methanol solution; Described pyrolysis solution is added in 5 milliliters of concentrated hydrochloric acids and is made by 95 milliliters of propyl carbinol;
1.4, the purifying of cyanidin
Cyanidin methanol solution, ethyl acetate and pure water by volume 1:2:2 mix, and under room temperature, extract, and collect water cornflowerblue cellulose solution; Water cornflowerblue cellulose solution and pure water by volume 1:10 mix, and through AB-8 macroporous resin adsorption, by the methanol-eluted fractions of 1 liter of concentration 20%, then use 100 milliliters of pure methanol-eluted fractions, collect red meoh eluate, obtain cornflowerblue cellulose solution;
1.5, the preparation of cyanidin sterling
Cornflowerblue cellulose solution is added to Rotary Evaporators, 40 ℃ of water-bath evaporations of temperature 10～30 minutes, Precipitation to be had, stops evaporation; Add 1 milliliter of concentrated hydrochloric acid, mix, place 1h at 4 ℃, 5000 revs/min of condition centrifugal collecting precipitates of rotating speed, Nitrogen evaporator dries up throw out, obtains the cyanidin powder that purity is greater than 98%; It is carmine that cyanidin powder is, and is insoluble in water, is soluble in methanol solution;
1.6, purifying obtains the efficient liquid phase chromatographic analysis of cyanidin sterling
Utilize high-efficient liquid phase chromatogram technology (HPLC) to detect the purity of the cyanidin that purifying obtains.
HPLC chromatographic condition: Japanese Shimadzu 20AD high performance liquid chromatograph; Chromatographic column: the dark 250*4.6 millimeter of the holy and pure blue 4u spoke of Féraud door (Phenomenex Synergi 4u Fusion 250*4.6mm); Detector: Japanese Shimadzu SPD6AV wavelengthtunable UV-detector; Sensitivity: 0.01; Detect wavelength: 530 nm; Flow velocity: 1.0 ml/min; Sample size: 5 microlitres; Adopt condition of gradient elution A phase: 1% acetic acid, B phase: trifluoroacetic acid aqueous solution, gradient be in first 20 minutes A by 90% to 87%, B by 10% to 13%; 20 minutes to 40 minutes A by 87% to 70%, B by 13% to 30%.
The inventive method has advantages of following several respects:
1, under natural condition, the cyanidin monomer of free state is seldom shown in, it is raw material that the present invention utilizes the Tea root widely of originating, and uses pycnogenols hydrolysis and purification technique to prepare high purity free state cyanidin monomer;
2, the method is simple to operate, does not need large-scale expensive laboratory apparatus, and organic solvent used is cheap, and recyclable, and cost is low;
3, present method can effectively be removed sugar, albumen, other types anthocyanogen and the flavonoid compound in extract, is applicable to obtaining high purity Minor centaury product.
Accompanying drawing explanation
Fig. 1 is the cyanidin two-dimensional highly effective liquid phase chromatographic figure obtaining after purifying.
Embodiment
Below in conjunction with accompanying drawing, by embodiment, the present invention is further described.
Embodiment:
Major equipment: 1. high performance liquid chromatograph (HPLC); 2. thermostat water bath; 3. Rotary Evaporators; 4. whizzer; 5. stamp mill; 6. AB-8 macroporous resin column; 7. Nitrogen evaporator.
Material and reagent:
1, detected material: Tea root (80 ℃ save backup);
2, main agents:
Acetone (analytical pure), acetic acid (analytical pure), methyl alcohol (analytical pure), propyl carbinol (analytical pure), ethyl acetate (analytical pure), concentrated hydrochloric acid (analytical pure).
Extracting solution: get 70 milliliters of acetone and add 29.5 ml waters and 0.5 milliliter of acetic acid to shake up.
Pyrolysis liquid: get 95 milliliters of propyl carbinols and add 5 milliliters of concentrated hydrochloric acids.
20% methyl alcohol: 20 ml methanol add in 80 ml waters and shake up.
The concrete operation step of preparation high purity cyanidin is as follows:
1), the extraction of pycnogenols
The Tea root that 20g is fresh is put into refiner, add 200 milliliters of extracting solutions, 1000 revs/min of rotating speeds, homogenate 5 minutes, collects homogenate; Under 4 ℃ of temperature, 5000 revs/min of conditions of rotating speed, centrifugation 10 minutes, collects supernatant liquor, obtains pycnogenols solution;
2), the purifying of pycnogenols
Pycnogenols solution and pure water by volume 1:10 mix, and through AB-8 macroporous resin adsorption, by the methanol-eluted fractions of 1 liter of concentration 20%, then use 100 milliliters of pure methanol-eluted fractions, collect meoh eluate, obtain pycnogenols crude product solution;
3), cyanidin methanol solution produces
Pycnogenols crude product solution and pyrolysis solution by volume 1:10 mix, and 95 ℃ of pyrolysis of temperature 1 hour, obtain cyanidin pyrolysis solution; Utilize Rotary Evaporators, temperature 70 C water-bath evaporation 1 hour, removes pyrolysis solution, and 10 milliliters of pure dissolve with methanol for residual dry powder, obtain cyanidin methanol solution;
4), the purifying of cyanidin
Cyanidin methanol solution, ethyl acetate and pure water by volume 1:2:2 mix, and under room temperature, extract, and collect water cornflowerblue cellulose solution; Water cornflowerblue cellulose solution and pure water by volume 1:10 mix, and through AB-8 macroporous resin adsorption, by the methanol-eluted fractions of 1 liter of concentration 20%, then use 100 milliliters of pure methanol-eluted fractions, collect red meoh eluate, obtain cornflowerblue cellulose solution;
5), the preparation of cyanidin sterling
Cornflowerblue cellulose solution is added to Rotary Evaporators, 40 ℃ of water-bath evaporations of temperature 10～30 minutes, Precipitation to be had, stops evaporation; 1 milliliter of the concentrated hydrochloric acid that adds concentration 36%, mixes, and places 1h at 4 ℃, 5000 revs/min of condition centrifugal collecting precipitates of rotating speed, and Nitrogen evaporator dries up throw out, obtains the cyanidin powder that purity is greater than 98%; It is carmine that the cyanidin powder of producing is, and is insoluble in water, is soluble in methanol solution.Cyanidin methanol solution is rose-red;
6), purifying obtains the efficient liquid phase chromatographic analysis of cyanidin sterling
Utilize high-efficient liquid phase chromatogram technology (HPLC) to detect the purity of the cyanidin that purifying obtains.
HPLC chromatographic condition: Japanese Shimadzu 20AD high performance liquid chromatograph; Chromatographic column: the dark 250*4.6 millimeter of the holy and pure blue 4u spoke of Féraud door (Phenomenex Synergi 4u Fusion 250*4.6mm); Detector: Japanese Shimadzu SPD6AV wavelengthtunable UV-detector; Sensitivity: 0.01; Detect wavelength: 530 nm; Flow velocity: 1.0 ml/min; Sample size: 5 microlitres; Adopt condition of gradient elution A phase: 1% acetic acid, B phase: trifluoroacetic acid aqueous solution, gradient be in first 20 minutes A by 90% to 87%, B by 10% to 13%; 20 minutes to 40 minutes A by 87% to 70%, B by 13% to 30%.
As seen from Figure 1, under 530nm wavelength, detect, only have the single chromatographic peak of cyanidin, without other assorted peaks, purity is greater than 98% relatively.
Claims (1)
1. a preparation method for high purity cyanidin, is characterized in that concrete operation step is as follows:
1.1, the extraction of pycnogenols
The Tea root that 20g is fresh is put into refiner, add 200 milliliters of extracting solutions, 1000 revs/min of rotating speeds, homogenate 5 minutes, collects homogenate; Under 4 ℃ of temperature, 5000 revs/min of conditions of rotating speed, centrifugation 10 minutes, collects supernatant liquor, obtains pycnogenols solution; Described extracting solution adds 29.5 ml waters and 0.5 milliliter of acetic acid to make by 70 milliliters of acetone;
1.2, the purifying of pycnogenols
Pycnogenols solution and pure water by volume 1:10 mix, and through AB-8 macroporous resin adsorption, by the methanol-eluted fractions of 1 liter of concentration 20%, then use 100 milliliters of pure methanol-eluted fractions, collect meoh eluate, obtain pycnogenols crude product solution;
1.3, cyanidin methanol solution produces
Pycnogenols crude product solution and pyrolysis solution by volume 1:10 mix, and 95 ℃ of pyrolysis of temperature 1 hour, obtain cyanidin pyrolysis solution; Utilize Rotary Evaporators, temperature 70 C water-bath evaporation 1 hour, removes pyrolysis solution, and 10 milliliters of pure dissolve with methanol for residual dry powder, obtain cyanidin methanol solution; Described pyrolysis solution is added in 5 milliliters of concentrated hydrochloric acids and is made by 95 milliliters of propyl carbinol;
1.4, the purifying of cyanidin
Cyanidin methanol solution, ethyl acetate and pure water by volume 1:2:2 mix, and under room temperature, extract, and collect water cornflowerblue cellulose solution; Water cornflowerblue cellulose solution and pure water by volume 1:10 mix, and through AB-8 macroporous resin adsorption, by the methanol-eluted fractions of 1 liter of concentration 20%, then use 100 milliliters of pure methanol-eluted fractions, collect red meoh eluate, obtain cornflowerblue cellulose solution;
1.5, the preparation of cyanidin sterling
Cornflowerblue cellulose solution is added to Rotary Evaporators, 40 ℃ of water-bath evaporations of temperature 10～30 minutes, Precipitation to be had, stops evaporation; Add 1 milliliter of concentrated hydrochloric acid, mix, place 1h at 4 ℃, 5000 revs/min of condition centrifugal collecting precipitates of rotating speed, Nitrogen evaporator dries up throw out, obtains the cyanidin powder that purity is greater than 98%; It is carmine that cyanidin powder is, and is insoluble in water, is soluble in methanol solution.
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| CN108586415A (en) * | 2018-04-18 | 2018-09-28 | 安徽农业大学 | The method for preparing high-purity cyanidin from violet cabbage |
| CN108586414A (en) * | 2018-04-18 | 2018-09-28 | 安徽农业大学 | The method for preparing high-purity delphinidin from purple eggplant |
| CN108484701A (en) * | 2018-05-14 | 2018-09-04 | 湖南农业大学 | A method of separation prepares tea tree purple bud anthocyanin high sterling |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1431205A (en) * | 2002-01-11 | 2003-07-23 | 华中农业大学 | Extraction agent used for extracting proanthocyanidin from plants and extracting method |
| CN101041678A (en) * | 2007-04-27 | 2007-09-26 | 浙江工商大学 | Method for separating preparation of corn-flower pigment-3-amylaceum glycosides from red bayberry |
| CN101353362A (en) * | 2008-04-15 | 2009-01-28 | 天津市尖峰天然产物研究开发有限公司 | Extraction method of cyaniding 3-0-glucoside |
| CN101380338A (en) * | 2007-08-01 | 2009-03-11 | 法鲁克斯控股股份有限公司 | Process of preparation of extracts rich in proanthocyanidins |
| WO2010142364A1 (en) * | 2009-06-09 | 2010-12-16 | Sartorius Stedim Biotech Gmbh | Method for obtaining secondary plant contents |
| CN102060829A (en) * | 2010-11-29 | 2011-05-18 | 中国科学院西北高原生物研究所 | Method for extracting anthocyanin from nitratia seeds |
| CN102532085A (en) * | 2011-12-29 | 2012-07-04 | 晨光生物科技集团股份有限公司 | Method for extracting proanthocyanidin from peanut coats |
-
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Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1431205A (en) * | 2002-01-11 | 2003-07-23 | 华中农业大学 | Extraction agent used for extracting proanthocyanidin from plants and extracting method |
| CN101041678A (en) * | 2007-04-27 | 2007-09-26 | 浙江工商大学 | Method for separating preparation of corn-flower pigment-3-amylaceum glycosides from red bayberry |
| CN101380338A (en) * | 2007-08-01 | 2009-03-11 | 法鲁克斯控股股份有限公司 | Process of preparation of extracts rich in proanthocyanidins |
| CN101353362A (en) * | 2008-04-15 | 2009-01-28 | 天津市尖峰天然产物研究开发有限公司 | Extraction method of cyaniding 3-0-glucoside |
| WO2010142364A1 (en) * | 2009-06-09 | 2010-12-16 | Sartorius Stedim Biotech Gmbh | Method for obtaining secondary plant contents |
| CN102060829A (en) * | 2010-11-29 | 2011-05-18 | 中国科学院西北高原生物研究所 | Method for extracting anthocyanin from nitratia seeds |
| CN102532085A (en) * | 2011-12-29 | 2012-07-04 | 晨光生物科技集团股份有限公司 | Method for extracting proanthocyanidin from peanut coats |
Non-Patent Citations (4)
| Title |
|---|
| Jian-Bin Li，等.Anthocyanins from red flowers of Camellia cultivar‘Dalicha’.《Phytochemistry》.2008,第69卷 * |
| 孟凡丽,等.红豆越橘果实中3-半乳糖-矢车菊色素的提取、分离及结构鉴定.《吉林农业大学学报》.2004,第26卷(第5期), * |
| 李春阳，等.葡萄籽原花青素结构单元的红外光谱分析.《食品与生物技术学报》.2005,第24卷(第4期), * |
| 肖付才,等.葡萄籽原花青素的提取和检测方法.《食品研究与开发》.2007,第28卷(第6期), * |
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