CN115286604A - Centrifugal fractional extraction to separate gallocatechin, epicatechin and caffeine from tea polyphenols - Google Patents
Centrifugal fractional extraction to separate gallocatechin, epicatechin and caffeine from tea polyphenols Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- 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
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/02—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6
- C07D473/04—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 two oxygen atoms
- C07D473/06—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 two oxygen atoms with radicals containing only hydrogen and carbon atoms, attached in position 1 or 3
- C07D473/12—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 two oxygen atoms with radicals containing only hydrogen and carbon atoms, attached in position 1 or 3 with methyl radicals in positions 1, 3, and 7, e.g. caffeine
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Abstract
The patent introduces a method for separating gallocatechin, epicatechin and caffeine from tea polyphenol by using a fractionation extraction technology, and no reaction extractant is added in the whole extraction process, and only physical distribution is involved. Has the advantages of low cost, strong operation continuity, simple operation flow, easy realization of industrial production and the like. The method utilizes the difference of distribution behaviors of various substances in the tea polyphenol in an organic phase and a water phase, realizes high mass transfer efficiency through the high-speed centrifugal action of the centrifugal extractor, and realizes the separation and purification of gallocatechin, epicatechin and caffeine in the tea polyphenol. Finally obtaining the gallocatechin in the water phase with the purity and yield of 83.7 percent and 95.8 percent respectively; the purity and yield of epicatechin obtained in the organic phase were 99.1% and 50.7%, respectively. The purity and yield of caffeine in the organic phase were 98.4% and 55.2%, respectively.
Description
Technical Field
The invention relates to a method for selectively separating epigallocatechin, epicatechin and caffeine from tea polyphenol by solvent extraction and multistage centrifugal fractionation extraction of a centrifugal extractor, which comprises the following steps: centrifugal fractional extraction method.
Background
Tea, as a beverage, is reputed to the world for its health benefits. Tea polyphenol is a main bioactive substance in tea, has various health-care effects, and is widely applied to the food industry. The tea extract contains tea polyphenols as main ingredient, is a compound of more than 30 kinds of phenolic compounds separated from natural plant tea, and comprises epigallocatechin gallate (EGCG), epigallocatechin (EGC), epicatechin gallate (ECG), epicatechin (EC), caffeine (CAF), epigallocatechin gallate (GCG), catechin (DL-C), etc., and many components in the extract have their own biological properties. The epigallocatechin has antioxidant, anticancer, and antimutagenic activities, and can stimulate bone formation and help slow bone aging; epicatechin has various physiological activities, such as reducing blood fat and weight, preventing cardiovascular and cerebrovascular diseases, preventing cancer, resisting mutation, etc. Compared with other catechins, the catechin compound can obviously improve the spatial memory and cognitive functions of middle-aged and elderly subjects, and arouses great interest of scholars at home and abroad.
The catechin is the most important functional active component of the tea, and the development of the green high-efficiency separation technology of the catechin monomer in the tea has remarkable economic and social benefits. At present, the technologies for separating catechin monomers mainly comprise chromatography, membrane separation, precipitation method and the like, the separation cost is high, the separation amount is small, the process is discontinuous, industrial amplification is difficult, and the market development of high-value-added functional active ingredients of tea leaves is limited. Meanwhile, a large amount of low-end tea leaves cannot be effectively utilized, and resource waste is caused. The development of research and application of a new green and efficient separation technology of catechin monomers can powerfully promote the deep development of tea resources in our province, and has very important strategic significance on the development of the tea industry in our province.
Aiming at the extraction of tea polyphenol in tea, a series of methods are developed at present, including solvent extraction, metal ion precipitation method and supercritical CO 2 Extraction methods, membrane separation methods, and the like. Tea polyphenol is low in price, is mainly used for food additives at present, and has no value given full play. In order to realize high-value utilization of tea polyphenol, catechin monomers in the tea polyphenol need to be further separated. In recent years, with the development of society and the evolution of technology, the extraction efficiency of catechin monomers and the quality of products need to be further improved. On the one hand, the product purity is required to be continuously improved, meanwhile, the production cost of unit products is continuously reduced, on the other hand, the product specification is diversified, namely, the tea polyphenol is subjected to differential purification and separation, and serial products with different specifications are obtained to meet the diversified market demands. The catechin monomers in tea polyphenols have similar structure and physicochemical properties, and are difficult to realize by conventional methodEfficient separation of bodies.
The prior method for purifying and refining catechin monomers mainly comprises the following steps: chromatography, supercritical fluid extraction, etc. The preparation chromatography is the most important preparation separation technology for obtaining high-purity natural active substances at present, the technology is rapidly developed in recent years, and the technologies such as column chromatography, high performance liquid chromatography, supercritical fluid chromatography, simulated moving bed chromatography, high-speed countercurrent chromatography and the like are developed, but most of the technologies are mainly used for preparing gallocatechin gallate and epicatechin gallate, but the technologies of epigallocatechin and epicatechin are relatively few, mainly comprise preparation type high performance liquid chromatography and high-speed countercurrent chromatography, belong to the separation range of laboratories, and the preparation amount is usually only a few milligrams, so that the industrial production cannot be realized. The content of caffeine in the tea is about 1% -2.8%, and the existing separation methods comprise a solvent method, an extraction sublimation method and a microwave-assisted method. However, high purity caffeine is difficult to obtain by a single extraction and still needs to be further purified. The main purification methods of caffeine include column chromatography, supercritical fluid extraction and biological enzymes. Column chromatography is currently the most common method for purifying caffeine, but does not meet the requirements of high purity and high volume production. Supercritical fluid extraction has high selectivity, safety and environmental protection, but has higher requirements on equipment. The biological enzyme method requires high storage conditions, and therefore, the cost for separation and purification is correspondingly high. These methods are not suitable for industrial continuous production.
The invention adopts a multi-stage centrifugal fractionation extraction technology to extract and separate gallocatechin, epicatechin and caffeine in tea polyphenol. According to the invention, according to different distribution behaviors of various substances in the tea polyphenol in a solvent system, the centrifugal action force of a centrifugal extractor is utilized to further enhance the mass transfer efficiency, and the epigallocatechin, the epicatechin and the caffeine are quickly and efficiently obtained. The technology overcomes the separation problems of other separation technologies, and has the characteristics of simple and convenient operation, high production efficiency, continuous large-scale production and high yield and purity.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for separating gallocatechin, epicatechin and caffeine from tea polyphenol, the method uses a green and efficient solvent system without a reaction extractant, utilizes a multi-stage centrifugal extraction separation technology, is simple and efficient in production process, is green and environment-friendly, meets the requirements of environmental protection and safety, and is suitable for industrial production of high-purity gallocatechin, epicatechin and caffeine.
The technical scheme provided by the invention is as follows: a method for continuously separating and purifying epigallocatechin, epicatechin and caffeine from tea polyphenol comprises the following steps:
(1) Dispensing experiments were performed. Dissolving tea polyphenol in water to serve as a water phase, and taking a pure organic solvent as an organic phase; mixing two phases with the same volume at a certain temperature, oscillating and standing, analyzing the concentrations of substances in aqueous phase and organic phase tea polyphenol by using a high performance liquid chromatograph, calculating the distribution ratio (monomer substance organic phase concentration/monomer substance water phase concentration) and separation factor of gallocatechin, epicatechin and caffeine in the tea polyphenol, and selecting a proper solvent system to perform a multistage centrifugal extraction experiment.
(2) A multi-stage centrifugal extraction experiment was performed. Dissolving a proper amount of tea polyphenol in water, and preparing a tea polyphenol water solution with a certain concentration as a feed liquid phase; taking the organic solvent system screened in the step (1) as an organic phase; taking pure water as a water phase, and carrying out a multi-stage centrifugal fractional extraction experiment. Wherein the volume flow ratio of the organic phase to the feed liquid phase is 0.1-10, the volume flow ratio of the organic phase to the water phase is 0.2-20, the feed liquid phase is at a feed position of 2-N-1, the temperature is controlled at 0-25 ℃, and N (N is more than or equal to 2 and less than or equal to 100) level centrifugal fractional extraction is carried out. The multi-stage centrifugal fractionation extraction experiment comprises the following steps: firstly, relatively large gallocatechin gallate, epigallocatechin gallate and epicatechin gallate are distributed in the tea polyphenol to an organic phase; taking the water phase containing epigallocatechin, epicatechin, caffeine and the like as a material liquid phase, performing fractional extraction for the second step, and remaining caffeine, catechin and impurities with relatively small distribution in the water phase to obtain an organic phase only containing epigallocatechin and epicatechin and a water phase containing caffeine; and distilling the organic phase under reduced pressure to remove the organic solvent, dissolving in water, performing experiment as the third step of fractionating the extracted liquid phase, and obtaining epigallocatechin in the water phase and epicatechin in the organic phase. Using the aqueous phase containing caffeine obtained in the second step as a feed liquid phase, removing catechin with partition coefficient higher than that of caffeine by fractional extraction, and performing fractional extraction in the second step to obtain an organic phase containing only caffeine.
(3) Distilling the water phase containing epigallocatechin, the organic phase containing epicatechin and the organic phase containing caffeine obtained in the step (2) under reduced pressure to obtain high-purity epigallocatechin, epicatechin and caffeine powder respectively.
The method is characterized in that: the method comprises the steps of firstly adopting a centrifugal fractional extraction method to selectively separate epigallocatechin, epicatechin and caffeine from tea polyphenol, selecting a proper extraction system, and utilizing the centrifugal acting force of a centrifugal extractor to realize rapid mixing and rapid separation of an aqueous phase and an organic phase according to different distribution behaviors of different monomer substances in the tea polyphenol in the aqueous phase and the organic phase so as to rapidly and efficiently obtain the epigallocatechin, epicatechin and caffeine with high purity. The purity and yield of epigallocatechin, epicatechin and caffeine can be greatly improved by multi-stage centrifugal fractional extraction. The method has the advantages of simple process, simple and convenient operation, stable product quality and suitability for large-scale production.
Description of the drawings:
FIG. 1 is a schematic diagram of a device used in a centrifugal extractor multi-stage fractionation extraction technology, wherein each serial number represents: (1) step (2) centrifugal fractionation extraction water phase inlet, step (2) centrifugal fractionation extraction water phase outlet, step (3) centrifugal fractionation extraction organic phase inlet, step (2) centrifugal fractionation extraction liquid phase inlet, and step (5) centrifugal fractionation extraction organic phase outlet. The centrifuges at the water phase inlet are numbered as 1 (1'), and the centrifuges are numbered in sequence according to the flow direction of the water phase, and each centrifuge forms 1 extraction stage. f denotes a feed position, stages 1 to f-1 constitute a washing section, and stages f to N constitute an extraction section.
[ detailed description ] embodiments
The invention is further illustrated below with reference to examples of the invention:
1. testing and analysis
Analysis of the yield and residual yield of the isolated product in the examples of the present invention was carried out using Agilent 1260 II HPLC (Agilent technologies, inc., agilent technologies, USA) and Inertsil ODS-3 column (250 mm X4.6 mm,5 μm, GL Sciences, japan).
2. Examples of the embodiments
Example 1
A solution having a concentration of 0.02 g/mL in water is taken as an aqueous phase, and an organic solvent (n-butanol, n-pentanol, isopentanol, n-hexanol, heptanol, n-octanol, ethyl acetate, butyl acetate, isobutyl acetate, n-hexane), a mixed solvent of ethyl acetate and n-hexane (mixed volume ratio: 8. Taking 5mL of each of the aqueous phase and the organic phase in a 50 mL centrifuge tube, fully shaking at 5 ℃ in a water bath constant temperature oscillator, standing for layering, calculating the distribution ratio (k) of each substance monomer in the tea polyphenol during equilibrium, wherein when butyl acetate is selected as an organic solvent, the distribution ratio of epigallocatechin is 0.934, the distribution ratio of epicatechin is 1.028, and the separation factors of substances adjacent to epigallocatechin and epicatechin are respectively 0.290 and 4.861. When the mixed solvent of ethyl acetate and n-hexane (8:1) is an organic phase, the partition ratio of epigallocatechin to epicatechin is 0.794 and 1.636, respectively. When a mixed solvent (1, v/v) of n-butyl alcohol and butyl acetate is used as an organic solvent, the distribution ratio of caffeine is 1.585, the distribution ratio of catechin is 6.640, and the separation factor between the two is 4.189. When n-butanol is the organic phase, the partition ratio of caffeine to impurities is 1.448 and 0.615, respectively, and the separation factor between them is 2.354.
Dissolving tea polyphenol in deionized water to prepare a 3L solution with the concentration of 0.02 g/mL as a feed liquid phase; butyl acetate solvent as the organic phase; taking deionized water as a water phase; connecting 10-stage centrifugal extractors in series, pumping the water phase into the centrifugal extractors by using a constant flow pump, pumping the organic phase into the centrifugal extractors from corresponding inlets when the water phase flows out from a raffinate phase outlet, and pumping the material liquid phase into the 3 rd-stage centrifugal extractors after the two-phase flow is stable for a period of time. The volume flow rate of the water phase is 5mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 5:2, and the volume flow rate ratio of the organic phase to the water phase is 5:3. The temperature is controlled to be 5 ℃ by circulating cooling water, a centrifugal fractional extraction experiment is carried out, and the extraction reaches the balance after 3 hours. By liquid phase detection and analysis, after extraction balance, the tea polyphenol with large proportion of gallocatechin gallate, epigallocatechin gallate and epicatechin gallate enter an organic phase, and other substances with small proportion enter a water phase; the yield of epigallocatechin in the aqueous phase was 89.8% and that of epicatechin was 81.3%.
Example 2
Dissolving tea polyphenol in deionized water to prepare a 3L solution with the concentration of 0.02 g/mL as a feed liquid phase; butyl acetate solvent as the organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the organic phase is pumped into the centrifugal extractors from corresponding inlets, and the material liquid phase is pumped into the 3 rd-stage centrifugal extractors after the two-phase flow is stable for a period of time. The volume flow rate of the water phase is 5mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 5:2, and the volume flow rate ratio of the organic phase to the water phase is 5:2. The temperature is controlled to be 5 ℃ by circulating cooling water, a centrifugal fractional extraction experiment is carried out, and the extraction reaches the balance after 3 hours. Through liquid phase detection and analysis, after extraction balance, the gallocatechin gallate, the epigallocatechin gallate and the epicatechin gallate with larger proportion in the tea polyphenol enter an organic phase, and other substances with smaller proportion enter a water phase; the yields of epigallocatechin and epicatechin in the aqueous phase were 89.8% and 89.2%, respectively.
Example 3
Taking the outlet water phase in example 2 as a feed liquid phase; butyl acetate solvent as the organic phase; taking deionized water as a water phase; and connecting 20-stage centrifugal extractors in series, pumping the water phase into the centrifugal extractors by using a constant flow pump, pumping the mixed organic solvent into the centrifugal extractors from corresponding inlets when the water phase flows out from a raffinate phase outlet, and pumping the liquid material liquid phase into the 6 th-stage centrifugal extractors after the two-phase flow is stable. The volume flow rate of the water phase is 2.5 mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 2.5. The extraction is carried out for 3 hours by centrifugal fractional distillation with circulating cooling water at 5 ℃ to ensure that the extraction is balanced. Through liquid phase detection and analysis, after extraction balance, the epigallocatechin and the epicatechin with larger distribution ratio in the liquid phase enter an organic phase, and other catechin, caffeine and other substances with smaller distribution ratio enter a water phase; an organic phase containing only epigallocatechin and epicatechin is obtained. The yields of epigallocatechin and epicatechin in the organic phase were 73.1% and 97.1%, respectively.
Example 4
Taking the organic phase only containing the epigallocatechin and the epicatechin in the example 3, distilling under reduced pressure to remove the organic solvent, and dissolving in water to obtain a water phase containing the epigallocatechin and the epicatechin as a feed liquid phase; a mixed solvent (8:1, v/v) of butyl acetate and n-hexane is used as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the mixed organic solvent is pumped into the centrifugal extractors from corresponding inlets, and the liquid material liquid phase is pumped into the 7 th-stage centrifugal extractors after the two-phase flow is stable. The volume flow rate of the water phase is 5mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 5:1, and the volume flow rate ratio of the organic phase to the water phase is 4:5. The temperature is controlled by circulating cooling water to carry out centrifugal fractional extraction for 3 hours at 5 ℃ so as to ensure that the extraction reaches the balance. Through liquid phase detection and analysis, after extraction balance, the epigallocatechin enters the water phase in the epicatechin organic phase in the feed liquid phase; the organic phase gave epicatechin with a purity of 99.1% and a yield of 50.7%.
Example 5
Taking the organic phase only containing the epigallocatechin and the epicatechin in the example 3, distilling under reduced pressure to remove the organic solvent, and dissolving in water to obtain a water phase containing the epigallocatechin and the epicatechin as a feed liquid phase; a mixed solvent (8:1, v/v) of butyl acetate and n-hexane is used as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the mixed organic solvent is pumped into the centrifugal extractors from corresponding inlets, and the liquid material liquid phase is pumped into the 7 th-stage centrifugal extractors after the two-phase flow is stable. The volume flow rate of the water phase is 5mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 5:1, and the volume flow rate ratio of the organic phase to the water phase is 1:1. The temperature is controlled by circulating cooling water to carry out centrifugal fractional extraction for 3 hours at 5 ℃ so as to ensure that the extraction reaches the balance. Detecting and analyzing by liquid phase, and after extraction balance, allowing epigallocatechin to enter into water phase in epicatechin organic phase in the feed liquid phase; the purity of the aqueous phase epigallocatechin was 83.7%, and the yield was 95.8%.
Example 6
Taking the organic phase only containing the epigallocatechin and the epicatechin in the example 3, distilling under reduced pressure to remove the organic solvent, and dissolving in water to obtain a water phase containing the epigallocatechin and the epicatechin as a feed liquid phase; a mixed solvent (8:1, v/v) of butyl acetate and n-hexane is used as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the mixed organic solvent is pumped into the centrifugal extractors from corresponding inlets, and the liquid material liquid phase is pumped into the 7 th-stage centrifugal extractors after the two-phase flow is stable. The volume flow of the water phase is 3mL/min, the volume flow ratio of the water phase to the feed liquid phase is 3:1, and the volume flow ratio of the organic phase to the water phase is 1:1. The extraction is carried out for 3 hours by centrifugal fractional distillation with circulating cooling water at 5 ℃ to ensure that the extraction is balanced. Through liquid phase detection and analysis, after extraction balance, the epigallocatechin enters the water phase in the epicatechin organic phase in the feed liquid phase; the purity and yield of epicatechin in the organic phase were 96.5% and 70.2%, respectively, while the purity and yield of epigallocatechin in the aqueous phase were 96.4% and 67.0%, respectively.
Example 7
Taking the outlet water phase containing the caffeine in the example 3 as a feed liquid phase; a mixed solvent of n-butanol and butyl acetate (1, v/v) solvent as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the organic phase is pumped into the centrifugal extractors from corresponding inlets, and the material liquid phase is pumped into the 5 th-stage centrifugal extractors after the two-phase flow is stable for a period of time. The volume flow rate of the water phase is 6 mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 3:2, and the volume flow rate ratio of the organic phase to the water phase is 1:3. The temperature is controlled to be 5 ℃ by circulating cooling water, a centrifugal fractional extraction experiment is carried out, and the extraction reaches the balance after 3 hours. Through liquid phase detection and analysis, after extraction balance, substances with a distribution ratio larger than that of caffeine in the water phase enter the organic phase, and other substances with a smaller distribution ratio enter the water phase; the yield of caffeine in the aqueous phase was 98.3%.
Example 8
Taking the outlet water phase in example 7 as a feed liquid phase; n-butanol as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the organic phase is pumped into the centrifugal extractors from corresponding inlets, and the material liquid phase is pumped into the 6 th-stage centrifugal extractors after the two-phase flow is stable for a period of time. The volume flow rate of the water phase is 2 mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 1:1, and the volume flow rate ratio of the organic phase to the water phase is 3:2. The temperature is controlled to be 5 ℃ by circulating cooling water, a centrifugal fractional extraction experiment is carried out, and the extraction reaches the balance after 3 hours. Through liquid phase detection and analysis, after extraction balance, more caffeine is distributed to enter an organic phase, and other substances with smaller distribution ratio enter a water phase; the yield and purity of caffeine were 72.1% and 98.0%, respectively.
Example 9
Taking the outlet water phase in example 7 as a feed liquid phase; n-butanol as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the organic phase is pumped into the centrifugal extractors from corresponding inlets, and the material liquid phase is pumped into the 6 th-stage centrifugal extractors after the two-phase flow is stable for a period of time. The volume flow rate of the water phase is 2 mL/min, the volume flow rate ratio of the water phase to the feed liquid is 1:1, and the volume flow rate ratio of the organic phase to the water phase is 1:1. The temperature is controlled to be 5 ℃ by circulating cooling water, a centrifugal fractional extraction experiment is carried out, and the extraction reaches the balance after 3 hours. Through liquid phase detection and analysis, after extraction balance, more caffeine is distributed to enter an organic phase, and other substances with smaller distribution ratio enter a water phase; the yield and purity of caffeine were 43.4% and 98.5%, respectively.
Example 10
Taking the outlet water phase in example 7 as a feed liquid phase; n-butanol as an organic phase; taking deionized water as a water phase; the 10-stage centrifugal extractors are connected in series, a constant flow pump is used for pumping the water phase into the centrifugal extractors, when the water phase flows out from the raffinate phase outlet, the organic phase is pumped into the centrifugal extractors from corresponding inlets, and the material liquid phase is pumped into the 6 th-stage centrifugal extractors after the two-phase flow is stable for a period of time. The volume flow rate of the water phase is 2 mL/min, the volume flow rate ratio of the water phase to the feed liquid phase is 2:1, and the volume flow rate ratio of the organic phase to the water phase is 1:1. The temperature is controlled to be 5 ℃ by circulating cooling water, a centrifugal fractional extraction experiment is carried out, and the extraction reaches the balance after 3 hours. Through liquid phase detection and analysis, after extraction balance, more caffeine is distributed to enter an organic phase, and other substances with smaller distribution ratio enter a water phase; the yield and purity of caffeine were 55.2% and 98.4%, respectively.
The above examples merely represent some embodiments of the present invention, which are described in more detail and in more detail, but are not to be construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent should be subject to the appended claims.
Claims (6)
1. The patent provides a method for separating tea polyphenol, namely Epigallocatechin (EGC) (1), epicatechin (EC) (2) and Caffeine (CAF) (3) by centrifugal fractionation and extraction,
(1) Epigallocatechin (EGC) (2) Epicatechin (EC) (3) Caffeine (CAF)
The method is realized by the device shown in the attached figure 1, and is characterized by comprising the following operation steps:
(1) Dissolving pre-purchased tea polyphenol with deionized water as a solvent to obtain a tea polyphenol water solution;
(2) Taking a tea polyphenol water solution as a water phase, taking an organic solvent as an organic phase, mixing the two phases for extraction and separation, fully oscillating, standing and sampling, analyzing by using high performance liquid chromatography to obtain the concentration of each substance in the balanced water phase and organic phase, calculating the distribution coefficient and separation factor of gallocatechin, epicatechin and caffeine in the tea polyphenol in an extraction system, and screening out a proper solvent system;
(3) Taking a proper amount of tea polyphenol to be dissolved in deionized water as a material liquid phase, taking the screened organic solvent as an organic phase, and taking the deionized water as a water phase;
(4) Pumping a water phase into an inlet (1) by using a metering pump, pumping an organic phase into an inlet (3) when the water phase flows out from an outlet (2), pumping a feed liquid phase into an inlet (4) after the organic phase flows out from an organic phase outlet (5), enabling the water phase and the organic phase to flow in a countercurrent manner, performing centrifugal fractional extraction with the N being more than or equal to 2 stages at a certain temperature, enabling the extraction process to reach a stable state, distributing substances with a distribution ratio being more than that of epigallocatechin and epicatechin to the organic phase to obtain a water phase containing epigallocatechin and epicatechin, taking the obtained water phase as the feed liquid phase, and performing centrifugal fractional extraction again; secondly, distributing epigallocatechin and epicatechin into an organic phase, removing an organic solvent from the organic phase by reduced pressure distillation and dissolving the organic phase in water, taking a water phase containing epigallocatechin and epicatechin as a material liquid phase, and carrying out fractional extraction in the third step; and thirdly, epigallocatechin is obtained in the water phase, and epicatechin is obtained in the organic phase. And (4) taking the outlet water phase in the second step as a feed liquid phase, and separating caffeine in the outlet water phase. Firstly, distributing substances with the distribution ratio larger than that of caffeine to an organic phase by one-step fractional extraction, continuously performing fractional extraction by using a water phase containing caffeine to obtain an organic phase only containing caffeine (5), and distilling the two-phase solution obtained in the step (4) under reduced pressure to obtain gallocatechin, epicatechin and caffeine powder.
2. The method of claim 1, wherein: the organic solvent is selected from n-butanol, n-pentanol, isoamyl alcohol, n-hexanol, heptanol, n-octanol, ethyl acetate, butyl acetate, isobutyl acetate, methyl tert-butyl ether and n-hexane.
3. The method of claim 1, wherein: selectively extracting gallocatechin, epicatechin and caffeine from tea polyphenols by using a multistage centrifugal extractor.
4. The method of claim 1, wherein: the molar ratio of the organic phase to the water phase in the multistage centrifugal fractional extraction is 0.2-20, and the molar ratio of the organic phase to the material liquid phase is 0.1-10.
5. The process of claim 1, wherein the feed liquid phase is fed at a level f (2. Ltoreq. F. Ltoreq.N-1).
6. The method of claim 1, wherein the temperature of the multi-stage centrifugal extraction process is controlled to be 0-25 ℃.
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