CN112852184A - Process for preparing high-purity black rice anthocyanin by secondary column chromatography and special chromatography device used in process - Google Patents
Process for preparing high-purity black rice anthocyanin by secondary column chromatography and special chromatography device used in process Download PDFInfo
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/12—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the preparation of the feed
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- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
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- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/22—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the construction of the column
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- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
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- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
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- B01D15/42—Selective adsorption, e.g. chromatography characterised by the development mode, e.g. by displacement or by elution
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- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
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Abstract
The invention discloses a process for preparing high-purity black rice anthocyanin by secondary column chromatography and a special chromatographic column device used in the process, wherein the process takes a common commercially available black rice extract with the anthocyanin content of 10-25% as a raw material, and the high-purity black rice anthocyanin dry powder is prepared by performing column chromatography purification, desorption solution concentration and spray drying on 200-mesh polyamide resin. The process and the special chromatographic column device can further purify and refine the common and commercially available low-purity black rice extract with the anthocyanin content of 10-25% to obtain the high-purity black rice anthocyanin with the anthocyanin content of 30-60%.
Description
Technical Field
The invention relates to a preparation method for further purifying a black rice extract by secondary column chromatography to obtain high-purity black rice anthocyanin, and belongs to the technical field of natural plant extraction.
Background
The black rice is a special variety formed by long-term cultivation of gramineous plant rice, is a treasure in rice, is vegetatively reputed as tribute rice, medicinal rice and longevity rice, and has special nutritional value. The black rice has high nutritive value and medicinal value, and is recorded in compendium of materia medica: has effects in nourishing yin, invigorating kidney, invigorating spleen, warming liver, improving eyesight, and promoting blood circulation. It has excellent curative effect on dizziness, anemia, leukotrichia, eye diseases, etc.
The color of black rice is different from that of other rice mainly because the outer skin layer contains natural plant pigment, namely anthocyanin, which has strong anti-aging effect. The total flavone substance in the black rice peel mainly comprises black rice anthocyanin.
The anthocyanin is a water-soluble natural pigment widely existing in plants in the nature, is also a main coloring matter in plant petals, is related to most of colorful colors of fruits, vegetables, flowers and the like, is widely existing in plants such as black rice, blueberries, bilberry, purple cabbage and the like, is a pure natural edible pigment with certain nutrition and pharmacological effects, and can resist various related diseases caused by free radicals of a human body. The health preserving and health care efficacy of anthocyanins is well known internationally and is known as "soft gold". In the beauty treatment world in western countries such as europe and the united states, anthocyanins are called "youth nutrition" and "oral cosmetics", and have been widely used for their antioxidant and radical-scavenging effects.
The black rice contains a large amount of anthocyanin, the marked component of the black rice extract is black rice anthocyanin, the content of the anthocyanin in the black rice extract produced in the domestic production and manufacturing industry is generally 10-25%, individual manufacturers can achieve the purity of the black rice anthocyanin close to 30%, the extraction method of the black rice extract has water extraction, alcohol extraction, enzymolysis, supercritical method and the like, but the purification and refining method is mainly based on macroporous resin column chromatography purification basically, when the black rice extract is used as a health-care functional food raw material or is added and used in the fields of medicines and cosmetics, the purity of the anthocyanin is important, the purity of the anthocyanin raw material used in the fields of foods and medicines in developed countries in Europe and America is generally more than 35%, and therefore, the preparation method of the high-purity black rice anthocyanin suitable for large-scale production has practicability and practical guiding significance. The application prospect of the high-purity black rice anthocyanin in the fields of health food, medicine and export trade of natural plant extracts is expected to be extremely wide.
Disclosure of Invention
The invention aims to provide a process for preparing high-purity black rice anthocyanin by secondary column chromatography and a special chromatography device used by the process. The invention takes a common commercially available black rice extract (the anthocyanin content is 10-25%) as a raw material, and prepares the high-purity black rice anthocyanin dry powder through polyamide column chromatography purification, desorption solution concentration and spray drying.
The process for preparing the high-purity black rice anthocyanin by the secondary column chromatography comprises the following steps:
1. column loading liquid pretreatment: according to the weight portion, the anthocyanin content of the commercially available black rice extract raw material is 10-25%, 10-15 portions of the commercially available black rice extract raw material are weighed, 100 portions of hydrochloric acid aqueous solution are fully stirred and dissolved at the temperature of 30-60 ℃, and the volume ratio concentration of the adopted hydrochloric acid aqueous solution is 2%.
2. Preparing a desorption solution: preparing an ethanol solution with the volume ratio of 60 percent, and adding 2 percent V/V hydrochloric acid.
3. The pressure of upper column liquid, washing liquid and desorption liquid is controlled to keep high and constant by adopting a chromatographic column device, and the high-pressure penetrating power is used for ensuring that feed liquid stably and smoothly penetrates through polyamide resin of the chromatographic column;
4. column loading and adsorption: and adsorbing the pretreated upper column liquid in the feed liquid storage tank 12 by polyamide resin in the chromatographic column 147 through an adsorption and desorption loop of the chromatographic column device, wherein the upper column pressure is set to be 5-7 kg, and the flow rate is controlled to be 2-2.5 column volumes per hour.
5. Washing with water: and after adsorption, washing the polyamide resin in the chromatographic column 147 with the washing liquid in the feed liquid storage tank 12 through an adsorption and desorption loop of the chromatographic column device, setting the upper column pressure to be 7-9 kg, controlling the flow rate to be 3-4 column volumes per hour, and washing until the eluent is clear.
6. Desorbing: after the water washing is finished, desorbing the polyamide resin in the chromatographic column 147 by using the desorption solution in the feed liquid storage tank 12 through an adsorption and desorption loop of a chromatographic column device, setting the column pressure of the desorption solution to be 5-7 kilograms, and controlling the flow rate to be 2.5-3 column volumes per hour to obtain the high-purity black rice anthocyanin desorption solution.
7. And (3) carrying out reduced pressure concentration, ethanol recovery and spray drying on the high-purity black rice anthocyanin desorption solution to obtain a dry powdery high-purity black rice anthocyanin finished product. The anthocyanin content of the high-purity black rice anthocyanin finished product is 30-60%.
8. Backwashing: the polyamide resin in the column 147 is backwashed by a backwash circuit of the column unit with alkaline and acidic backwash liquids alternately to restore the adsorption function of the polyamide resin.
The special chromatographic device used in the process of preparing the high-purity black rice anthocyanin by the secondary column chromatography comprises a constant-pressure water supply system, a feed liquid storage tank and a chromatographic column device, wherein a feed inlet of the constant-pressure water supply system is communicated with the feed liquid storage tank, an outlet of the constant-pressure water supply system is communicated with a first interface of the chromatographic column device through a first pipeline, a first valve is arranged between the first pipeline and the first interface, a third interface of the chromatographic column device is communicated with a liquid discharge pipe, the liquid discharge pipe is provided with a fourth valve, the first pipeline is communicated with the liquid discharge pipe at an inlet of the fourth valve through a second valve, a third valve and a second pipeline, and a second interface of the chromatographic column device is communicated with the liquid discharge pipe through a fifth valve, a third pipeline and a sixth valve;
the first pipeline, the first valve, the first interface, the chromatographic column device, the third interface, the fourth valve and the liquid discharge pipe form an adsorption and desorption loop;
the first pipeline, the second valve, the second pipeline, the third valve, the third interface, the chromatographic column device, the second interface, the fifth valve, the third pipeline, the sixth valve and the liquid discharge pipe form a back washing loop.
The chromatography column device comprises an enclosure, a first flange, a second flange, a baffle, a screen, a cylinder body, chromatography columns and end plates, wherein the end plates are arranged at two ends of the cylinder body, the peripheral surface of each end plate and the inner wall of the cylinder body are in a sealed state, a plurality of chromatography columns are tightly arranged in the cylinder body and penetrate through the end plates, the peripheral surface of each chromatography column and the hole wall of each end plate are in a sealed state, the first flange is welded at two ends of the cylinder body in a sealed mode, the second flange is welded at the enclosure at two ends of the cylinder body in a sealed mode, the first flange and the second flange are fixed together through bolts and nuts, the baffle and the screen are clamped between the first flange and the second flange, the screen is located on the inner side, and a; a first connector and a second connector are arranged on the sealing cover on the upper part of the cylinder body; a third interface is arranged on the sealing cover at the lower part of the cylinder body; the chromatographic column is filled with 200-mesh polyamide resin by a wet method.
The diameter-height ratio of the chromatographic column is 1: 12.
the metal parts of the special chromatographic device are all made of stainless steel.
The working process and principle of the special chromatographic device are as follows:
the constant pressure water supply system keeps the pressure of the upper column liquid, the washing liquid and the desorption liquid high and constant, ensures that the feed liquid stably and smoothly permeates through the polyamide resin of the chromatographic column by using high-pressure penetrating power,
1. column loading and adsorption: the constant pressure water supply system absorbs the pretreated upper column liquid in the feed liquid storage tank through the polyamide resin in the chromatographic column through the adsorption and desorption loops of the chromatographic column device.
2. Washing with water: after the adsorption is finished, the constant-pressure water supply system washes the water washing liquid in the feed liquid storage tank through an adsorption and desorption loop of the chromatographic column device to wash the polyamide resin in the chromatographic column.
3. Desorbing: after the water washing is finished, the constant-pressure water supply system desorbs polyamide resin in the chromatographic column from the desorption solution in the feed liquid storage tank through an adsorption and desorption loop of the chromatographic column device, and the high-purity black rice anthocyanin desorption solution is obtained after desorption.
4. Backwashing: the polyamide resin in the column 147 is backwashed by a backwash circuit of the column unit with alkaline and acidic backwash liquids alternately to restore the adsorption function of the polyamide resin.
The method uses the low-purity black rice extract as a raw material, performs secondary column chromatography purification through a 200-mesh polyamide resin column, and prepares the high-purity black rice anthocyanin dry powder by utilizing the principle that polyamide resin and anthocyanin molecules are selectively adsorbed by hydrogen bond combination, concentrating desorption solution and spray drying.
The invention has the beneficial effects that:
1. polyamide resin is the most ideal adsorbent for flavonoids, is mainly used for the subdivision and purification of flavonoids, and is different from the adsorption principle of macroporous resin, wherein the polyamide resin belongs to the chemical adsorption principle by virtue of hydrogen bond adsorption, the macroporous resin is the physical adsorption principle, and the finer the resin is in chromatographic application, the better the separation effect is. Therefore, the separation and purification of the black rice extract by using 200-mesh polyamide resin secondary column chromatography is a fundamental guarantee for preparing high-purity anthocyanin, and the low-purity black rice extract with the anthocyanin content of 10-25% in the common market can be further purified and refined to obtain the high-purity black rice anthocyanin with the anthocyanin content of 30-60%.
2. The application obstacle of the polyamide resin in the large-scale production is the flow velocity problem caused by the resin fineness, the flow velocity is slowed when the resin fineness exceeds 60 meshes, the permeability is deteriorated, the resin fineness exceeds 100 meshes, the flow velocity on a column is nearly stopped under normal pressure, and the production period is too long, so that the large-scale production and manufacture cannot be adapted to, therefore, in the actual production, manufacturers mostly select macroporous resin for purification, which is a problem of no choice, one of the innovations of the invention is to solve the application obstacle of the polyamide resin with the particle size of more than 100 meshes in the large-scale production, control the pressure of upper column liquid, washing liquid and desorption liquid to be kept high and constant by adopting a chromatographic column device, ensure that the feed liquid stably and smoothly penetrates through the polyamide resin by using high-pressure penetrating power, so that the flow. The constant-pressure water supply system adopting the chromatographic column device adopts the prior art, and the control principle is that a high-pressure stainless steel pump is connected with the variable-frequency control of an intelligent constant-pressure control cabinet.
3. In the application of the polyamide resin in column chromatography, the separation and purification effect is better as the fineness of the resin is smaller, and the separation and purification effect is better as the diameter-height ratio is larger, the diameter-height ratio is generally required to be more than 1:8, the laboratory can achieve 1:50, but in actual large-scale production, the separation and purification effect is better as the diameter-height ratio is larger, the height of a chromatographic column is about 3 meters, the cost performance is the highest, the operation is most convenient for workers, and one of the innovativeness of the invention is to ensure the practicability and the economy of large-scale production. Therefore, the chromatographic column adopts a combination mode that stainless steel is welded to manufacture small columns (the diameter-height ratio of the small columns is 1: 12) with the diameter of 0.2 meter and the height of 2.4 meters, and the small columns are assembled in a large column with the diameter of 1 meter or 0.6 meter in a bundling manner, 19 small columns can be assembled in the large column with the diameter of 1 meter, and 7 small columns can be assembled in the large column with the diameter of 0.6 meter. The advanced performance of the system is that the operation of a plurality of small chromatographic columns is met by one set of variable frequency pump feeding system of the intelligent constant-pressure control cabinet on the premise of ensuring the radial-height ratio, the space usage is saved, the labor is saved, and the auxiliary facilities are saved.
4. In the pretreatment of the upper column liquid, hydrochloric acid is added, because the anthocyanin has better solubility and stability in an acid solvent, the anthocyanin is heated to 30-60 ℃ for quick dissolution, and the anthocyanin is easy to be adsorbed by resin when being loaded on the column at high temperature.
5. In the preparation of the desorption solution, the purpose of adopting acid preparation is to facilitate desorption and ensure the stability of the anthocyanin.
Drawings
FIG. 1 is a schematic diagram of the structure of a special chromatographic device used in the process of the invention.
FIG. 2 is a cross-sectional view of a chromatography column of the dedicated chromatography device.
FIG. 3 is an end view of a chromatography column of the dedicated chromatography device.
Detailed Description
The first embodiment is as follows:
with reference to fig. 1, 2 and 3, the process for preparing high-purity black rice anthocyanin by secondary column chromatography comprises the following steps:
1. column loading liquid pretreatment: according to the weight portion, the anthocyanin content of the commercially available black rice extract raw material is 20%, 10 portions of the commercially available black rice extract raw material are weighed, 100 portions of hydrochloric acid aqueous solution are fully stirred and dissolved at the temperature of 30-60 ℃, and the volume ratio concentration of the adopted hydrochloric acid aqueous solution is 2%.
2. Preparing a desorption solution: preparing an ethanol solution with the volume ratio of 60 percent, and adding 2 percent V/V hydrochloric acid.
3. The special chromatographic device is adopted to control the pressure of the upper column liquid, the washing liquid and the desorption liquid to keep high and constant, and the high-pressure penetrating power is used for ensuring that the feed liquid stably and smoothly penetrates through the polyamide resin of the chromatographic column; the diameter of the chromatographic column 147 of the chromatographic column device is 0.2 meter, the height of the chromatographic column 147 is 2.4 meters, the number of the chromatographic columns 147 is 19, and the inner diameter of the cylinder 146 is 1 meter; the chromatographic column 147 is wet packed with 200 mesh polyamide resin.
4. Column loading and adsorption: and adsorbing the pretreated upper column liquid in the feed liquid storage tank 12 by polyamide resin in the chromatographic column 147 through an adsorption and desorption loop of the chromatographic column device, wherein the upper column pressure is set to be 5-7 kg, and the flow rate is controlled to be 2-2.5 column volumes per hour.
5. Washing with water: and after adsorption, washing the polyamide resin in the chromatographic column 147 with the washing liquid in the feed liquid storage tank 12 through an adsorption and desorption loop of the chromatographic column device, setting the upper column pressure to be 7-9 kg, controlling the flow rate to be 3-4 column volumes per hour, and washing until the eluent is clear.
6. Desorbing: after the water washing is finished, desorbing the polyamide resin in the chromatographic column 147 by using the desorption solution in the feed liquid storage tank 12 through an adsorption and desorption loop of a chromatographic column device, setting the column pressure of the desorption solution to be 5-7 kilograms, and controlling the flow rate to be 2.5-3 column volumes per hour to obtain the high-purity black rice anthocyanin desorption solution.
7. Concentrating the high-purity black rice anthocyanin desorption solution under reduced pressure, recovering ethanol, and spray drying to obtain a dry powdery high-purity black rice anthocyanin finished product; the anthocyanin content of the high-purity black rice anthocyanin finished product is 30-60%.
8. Backwashing: the polyamide resin in the column 147 is backwashed by a backwash circuit of the column unit with alkaline and acidic backwash liquids alternately to restore the adsorption function of the polyamide resin.
Example two:
with reference to fig. 1, 2 and 3, the process for preparing high-purity black rice anthocyanin by secondary column chromatography comprises the following steps:
1. column loading liquid pretreatment: the anthocyanin content of the commercially available black rice extract raw material is 20% in parts by weight, 15 parts of the commercially available black rice extract raw material is weighed, and is fully stirred and dissolved at the temperature of 30-60 ℃ by 100 parts of hydrochloric acid aqueous solution, and the volume ratio concentration of the adopted hydrochloric acid aqueous solution is 2%.
2. Preparing a desorption solution: preparing an ethanol solution with the volume ratio of 60 percent, and adding 2 percent V/V hydrochloric acid.
3. The special chromatographic device is adopted to control the pressure of the upper column liquid, the washing liquid and the desorption liquid to keep high and constant, and the high-pressure penetrating power is used for ensuring that the feed liquid stably and smoothly penetrates through the polyamide resin of the chromatographic column; the diameter of the chromatographic column 147 of the chromatographic column device is 0.2 meter, the height of the chromatographic column 147 is 2.4 meters, the number of the chromatographic columns 147 is 19, and the inner diameter of the cylinder 146 is 1 meter; the chromatographic column 147 is wet packed with 200 mesh polyamide resin.
4. Column loading and adsorption: and adsorbing the pretreated upper column liquid in the feed liquid storage tank 12 by polyamide resin in the chromatographic column 147 through an adsorption and desorption loop of the chromatographic column device, wherein the upper column pressure is set to be 5-7 kg, and the flow rate is controlled to be 2-2.5 column volumes per hour.
5. Washing with water: and after adsorption, washing the polyamide resin in the chromatographic column 147 with the washing liquid in the feed liquid storage tank 12 through an adsorption and desorption loop of the chromatographic column device, setting the upper column pressure to be 7-9 kg, controlling the flow rate to be 3-4 column volumes per hour, and washing until the eluent is clear.
6. Desorbing: after the water washing is finished, desorbing the polyamide resin in the chromatographic column 147 by using the desorption solution in the feed liquid storage tank 12 through an adsorption and desorption loop of a chromatographic column device, setting the column pressure of the desorption solution to be 5-7 kilograms, and controlling the flow rate to be 2.5-3 column volumes per hour to obtain the high-purity black rice anthocyanin desorption solution.
7. And (3) carrying out reduced pressure concentration, ethanol recovery and spray drying on the high-purity black rice anthocyanin desorption solution to obtain a dry powdery high-purity black rice anthocyanin finished product. The anthocyanin content of the high-purity black rice anthocyanin finished product is 30-60%.
8. Backwashing: the polyamide resin in the column 147 is backwashed by a backwash circuit of the column unit with alkaline and acidic backwash liquids alternately to restore the adsorption function of the polyamide resin.
As shown in fig. 1, fig. 2 and fig. 3, the special chromatography device for the process of preparing high-purity black rice anthocyanin by secondary column chromatography, the special chromatography device 1 comprises a constant-pressure water supply system 11, a feed liquid storage tank 12 and a chromatography column device 14, a feed inlet of the constant-pressure water supply system 11 is communicated with the feed liquid storage tank 12, an outlet of the constant-pressure water supply system 11 is communicated with a first interface 15 of the chromatography column device 14 through a first pipeline 13, a first valve 21 is arranged between the first pipeline 13 and the first interface 15, a third interface 17 of the chromatography column device 14 is communicated with a liquid discharge pipe 18, the liquid discharge pipe 18 is provided with a fourth valve 24, the first pipeline 13 is communicated with a second valve 22, the third valve 23 and the second pipeline 19 are communicated with the drain pipe 18 at the inlet of the fourth valve 24, and the second interface 16 of the chromatographic column device 14 is communicated with the drain pipe 18 through a fifth valve 25, a third pipeline 20 and a sixth valve 26;
the first pipeline 13, the first valve 21, the first interface 15, the chromatographic column device 14, the third interface 17, the fourth valve 24 and the liquid discharge pipe 18 form an adsorption and desorption loop;
the first line 13, the second valve 22, the second line 19, the third valve 23, the third interface 17, the chromatography column device 14, the second interface 16, the fifth valve 25, the third line 20, the sixth valve 26 and the drain 18 constitute a back-flushing circuit.
The chromatographic column device 14 is composed of an enclosure 141, a first flange 142, a second flange 143, a baffle 144, a screen 145, a cylinder 146, chromatographic columns 147 and end plates 148, wherein the end plates 148 are arranged at two ends of the cylinder 146, the peripheral surface of the end plate 148 and the inner wall of the cylinder 146 are in a sealed state, the cylinder 146 is internally and tightly provided with the plurality of chromatographic columns 147, the plurality of chromatographic columns 147 penetrate through the end plates 148, the peripheral surface of each chromatographic column 147 and the hole wall of the end plate 148 are in a sealed state, the two ends of the cylinder 146 are hermetically welded with the first flange 142, the enclosure 141 at the two ends of the cylinder 146 is hermetically welded with the second flange 143, the first flange 142 and the second flange 143 are fixed together through bolts and nuts, the baffle 144 and the screen 145 are clamped between the first flange 142 and the second flange 143, the screen 14 is positioned at the inner side, and the baffle 144 is provided with; the enclosure 141 on the upper part of the cylinder 146 is provided with a first interface 15 and a second interface 16; the enclosure 141 at the lower part of the cylinder 146 is provided with a third interface 17; the chromatographic column 147 is wet packed with 200 mesh polyamide resin.
The aspect ratio of the column 147 is 1: 12.
the metal parts of the special chromatographic device 1 are all made of stainless steel.
In this embodiment, the diameter of the chromatography column 147 is 0.2 m, the height of the chromatography column 147 is 2.4 m, the number of the chromatography columns 147 is 19, and the inner diameter of the cylinder 146 is 1 m; or the number of the chromatography columns 147 is 7, and the inner diameter of the cylinder 146 is 0.6 m.
The working process and principle of the special chromatographic device are as follows:
the constant pressure water supply system 11 keeps the pressure of the upper column liquid, the washing liquid and the desorption liquid high and constant, ensures that the feed liquid stably and smoothly permeates through the polyamide resin of the chromatographic column by using high-pressure penetrating power,
1. column loading and adsorption: the constant pressure water supply system 11 makes the pretreated upper column liquid in the feed liquid storage tank 12 pass through the adsorption and desorption loop of the chromatographic column device and be adsorbed by the polyamide resin in the chromatographic column 147.
2. Washing with water: after the adsorption is finished, the constant pressure water supply system 11 washes the polyamide resin in the chromatographic column 147 with the washing liquid in the feed liquid storage tank 12 through the adsorption and desorption loops of the chromatographic column device.
3. Desorbing: after the water washing is finished, the constant-pressure water supply system 11 desorbs the polyamide resin in the chromatographic column 147 from the desorption solution in the feed solution storage tank 12 through the adsorption and desorption loops of the chromatographic column device, and the high-purity black rice anthocyanin desorption solution is obtained after desorption.
4. Backwashing: the polyamide resin in the column 147 is backwashed by means of a backwash circuit of the column unit with alkaline and acidic backwash liquids to restore the adsorption function of the polyamide resin.
Claims (5)
1. The process for preparing the high-purity black rice anthocyanin by the secondary column chromatography is characterized by comprising the following steps of: the method comprises the following steps:
1) and column liquid pretreatment: weighing 10-15 parts of commercially available black rice extract raw material according to the weight part of the anthocyanin content of the commercially available black rice extract raw material, and fully stirring and dissolving 100 parts of hydrochloric acid aqueous solution at the temperature of 30-60 ℃, wherein the volume ratio concentration of the adopted hydrochloric acid aqueous solution is 2%;
2) and preparing desorption liquid: preparing an ethanol solution with the volume ratio of 60%, and adding 2% V/V hydrochloric acid;
3) the special chromatographic device is adopted to control the pressure of the upper column liquid, the water washing liquid and the desorption liquid to keep high and constant, and the high-pressure penetrating power is used for ensuring that the feed liquid stably and smoothly penetrates through the polyamide resin of the chromatographic column;
4) and column loading adsorption: adsorbing pretreated upper column liquid in a feed liquid storage tank (12) by polyamide resin in a chromatographic column (147) through an adsorption and desorption loop of a chromatographic device, wherein the upper column pressure is set to be 5-7 kilograms, and the flow rate is controlled to be 2-2.5 column volumes per hour;
5) and washing with water: after adsorption, washing the polyamide resin in the chromatographic column (147) with water from the water washing liquid in the feed liquid storage tank (12) through an adsorption and desorption loop of the chromatographic device, setting the upper column pressure to be 7-9 kg, controlling the flow rate to be 3-4 column volumes per hour, and washing until the eluent is clear;
6) and desorption: after the water washing is finished, desorbing polyamide resin in the chromatographic column (147) by using a desorption solution in the feed liquid storage tank (12) through an adsorption and desorption loop of a chromatographic device, setting the column pressure of the desorption solution to be 5-7 kg, and controlling the flow rate to be 2.5-3 column volumes per hour to obtain a high-purity black rice anthocyanin desorption solution;
7) concentrating the high-purity black rice anthocyanin desorption solution under reduced pressure, recovering ethanol, and spray drying to obtain a dry powdery high-purity black rice anthocyanin finished product;
8) and backwashing: the polyamide resin in the column 147 is back-flushed through the back-flushing circuit of the chromatography apparatus with alkaline and acidic back-flushing liquids alternately to restore the adsorption function of the polyamide resin.
2. The special chromatographic device for the process of preparing the high-purity black rice anthocyanin by the secondary column chromatography in claim 1 is characterized in that: the special chromatographic device (1) comprises a constant-pressure water supply system (11), a feed liquid storage tank (12) and a chromatographic device (14), wherein a feed inlet of the constant-pressure water supply system (11) is communicated with the feed liquid storage tank (12), an outlet of the constant-pressure water supply system (11) is communicated with a first interface (15) of the chromatographic device (14) through a first pipeline (13), a first valve (21) is arranged between the first pipeline (13) and the first interface (15), a third interface (17) of the chromatographic device (14) is communicated with a liquid discharge pipe (18), the liquid discharge pipe (18) is provided with a fourth valve (24), the first pipeline (13) is communicated with the liquid discharge pipe (18) at an inlet of the fourth valve (24) through a second valve (22), a third valve (23) and a second pipeline (19), the second interface (16) of the chromatography device (14) is communicated with the drain pipe (18) through a fifth valve (25), a third pipeline (20) and a sixth valve (26);
the first pipeline (13), the first valve (21), the first interface (15), the chromatography device (14), the third interface (17), the fourth valve (24) and the liquid discharge pipe (18) form an adsorption and desorption loop;
the first pipeline (13), the second valve (22), the second pipeline (19), the third valve (23), the third interface (17), the chromatography device (14), the second interface (16), the fifth valve (25), the third pipeline (20), the sixth valve (26) and the liquid discharge pipe (18) form a back washing loop.
The chromatography device (14) is composed of an enclosure (141), a first flange (142), a second flange (143), a baffle (144), a screen (145), a cylinder (146), chromatography columns (147) and end plates (148), wherein the end plates (148) are arranged at two ends of the cylinder (146), the peripheral surface of each end plate (148) is in a sealing state with the inner wall of the cylinder (146), a plurality of chromatography columns (147) are closely arranged in the cylinder (146), the plurality of chromatography columns (147) penetrate through the end plates (148), the peripheral surface of each chromatography column (147) is in a sealing state with the hole walls of the end plates (148), the first flange (142) is hermetically welded at two ends of the cylinder (146), the second flange (143) is hermetically welded at the enclosure (141) at two ends of the cylinder (146), the first flange (142) and the second flange (143) are fixed together through bolts and nuts, the baffle (144) and the screen (145) are clamped between the first flange (142) and the second flange (143), the screen is positioned at the inner side of the screen 14, and the baffle (144) is provided with a plurality of through holes (1441); the enclosure (141) at the upper part of the cylinder body (146) is provided with a first interface (15) and a second interface (16); the enclosure (141) at the lower part of the cylinder body (146) is provided with a third interface (17); the chromatographic column (147) is packed with 200 mesh polyamide resin by wet method.
3. The special chromatographic device for the process of preparing the high-purity black rice anthocyanin by the secondary column chromatography in claim 2 is characterized in that: the diameter-height ratio of the chromatographic column (147) is 1: 12.
4. the special chromatographic device for the process of preparing the high-purity black rice anthocyanin by the secondary column chromatography in claim 2 is characterized in that: the metal parts of the special chromatographic device (1) are all made of stainless steel.
5. The special chromatographic device for the process of preparing the high-purity black rice anthocyanin by the secondary column chromatography in claim 3 is characterized in that: the diameter of the chromatographic column (147) is 0.2 m, the height of the chromatographic column (147) is 2.4 m, the number of the chromatographic columns (147) is 19, and the inner diameter of the cylinder body (146) is 1 m; or the diameter of the chromatographic column (147) is 0.2 m, the height of the chromatographic column (147) is 2.4 m, the number of the chromatographic columns (147) is 7, and the inner diameter of the cylinder body (146) is 0.6 m.
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