CN103242314A - Method for comprehensively extracting effective ingredients in tea - Google Patents
Method for comprehensively extracting effective ingredients in tea Download PDFInfo
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Abstract
The invention relates to a method for comprehensively extracting effective ingredients in tea. The method comprises the following steps of: with a faintly acidic water solution as a main extraction solvent and a certain amount of organic solvent as an auxiliary extraction solvent, carrying out step-by-step extraction on water extracts to obtain four effective ingredients, namely tea polyphenol, caffeine, tea polysaccharide and theanine; and meanwhile, extracting tea protein from tea residues by using an alkali dissolution and acid deposition method. According to the method, as water is used as the main solvent, the cost is low, the reactions are mild and active ingredients are favorably preserved; as a certain amount of organic solvent is used as the auxiliary extraction solvent, the problems of overhigh cost and product pollution caused by use of a large amount of organic solvents are solved; meanwhile, five effective ingredients, namely the tea polyphenol, the tea polysaccharide, the caffeine, the theanine and the tea protein are extracted, and therefore high extraction rate, high purity and high utilization rate of tea are achieved; the solvents are recycled in the extraction process, so that the preparation cost is reduced; and no alkali and acid waste liquor is discharged in the overall process, and therefore the method is environment-friendly. The overall preparation flow is high in efficiency and low in cost, and therefore the method has a favorable application value.
Description
Technical field
The invention belongs to field of deep tea processing technology, relate to the method for effective constituent in the efficient production tealeaves of the method, particularly a kind of suitable suitability for industrialized production of effective constituent in a kind of comprehensive extraction tealeaves.
Background technology
As the Traditional health care beverage of the Chinese nation, tealeaves just is subjected to liking of people since ancient times and payes attention to.Moreover, in the world today, tealeaves has become first of the three big beverages (tealeaves, coffee, cocoa).China is the quite abundant country of a tea resources, yet along with superior day by day, a large amount of low and middle-grade tealeaves deadstocks of tealeaves consumption, can produce a large amount of byproducts simultaneously in the Tea Production course of processing, and utilization ratio is extremely low.Therefore, utilize the tankage of low and middle-grade tealeaves and Tea Processing to extract its effective constituent, greatly develop deep tea processing technology, open up the new purposes of tealeaves, not only can open up low and middle-grade tea market, take full advantage of tea resources, also will become the new direction of China's tealeaves industry development.
Scientific research and clinical medicine experiment show that tealeaves has many health-care effects, and this is inseparable with the effective efficiency composition in the tealeaves.Effective constituent in the tealeaves comprises tea-polyphenol, tea polysaccharide, trimethyl-xanthine, theanine, tea albumen etc.Tea-polyphenol is not only a kind of natural antioxidant, and have reducing blood-fat, a hypotensive and reducing cholesterol, arteriosclerosis, anti-mutation, anticancer change, anti-ageing, radioprotective, physiological function such as antiviral have purposes widely in industries such as medicine, beverage, food, health care; Tea polysaccharide has excellent pharmacological function, experiment shows, after tea polysaccharide enters human body, but antithrombotic, anticoagulation, radioprotective, reducing blood-fat, hypoglycemic delay senility, and can strengthen the non-specific immunity of human body at short notice, control Tiroidina hypertrophy and diabetes are improved hemopoietic function etc.; Trimethyl-xanthine has cardiac stimulus, excited cerebral nerve and diuretic properties as important medical material, also is the additive of multiple beverage simultaneously; Theanine has step-down, improves memory, strengthens the effects such as ability that human body is resisted virus infection, is medicine and the healthcare products composition of a kind of novel antiviral, bacterium, fungi; Tea albumen can be removed superoxide anion, has reducing blood-fat, and effects such as anti-mutation, atherosclerosis and coronary heart disease are protein a kind of not only nutritious but also that have plurality of health care functions.Therefore, high efficiency extraction tea-polyphenol from tealeaves, tea polysaccharide, trimethyl-xanthine, effective constituents such as theanine and tea albumen can produce great economic benefit and social benefit.
The method of extracts active ingredients mainly contains water extract method in the tealeaves, organic solvent extraction, the precipitator method, supercritical CO
2Extraction process, ultrasonic extraction method and microwave extraction method etc.Though the organic solvent extraction extraction yield is higher, cause organic solvent to pollute easily; Precipitator method three-waste pollution is serious, has food safety hidden danger; Supercritical CO
2Extraction process is to the equipment requirements height, and device is complicated, involves great expense; Ultrasonic extraction method and microwave extraction method energy consumption are big, and how the cost height is as supplementary means.Therefore, water extract method is both economical method for extracting effective content of tea efficiently.
At present, relevant comprehensively the patent report of extraction plurality of active ingredients is less from tealeaves, as: Chinese patent (96113134.9) has been reported a kind of method of extracting tea-polyphenol and byproduct thereof from tealeaves, the technology relative complex, and energy consumption is higher; Chinese patent (02132857.9) has been reported a kind of cold extraction method of effective content of tea, obtain effective constituent extracting solution and not with each component separating; Chinese patent application (200410015905.2) has reported that a kind of microwave-assisted extracts the method for tea-polyphenol, tea polysaccharide, trimethyl-xanthine, and the purity of tea-polyphenol is 80% in this production technique, but yield only is 5%; Chinese patent application (200410089179.9) has been reported the method for effective constituent in a kind of ultrasonic concussion extract at low temperature tealeaves, though extraction time is short, and the energy consumption height, the yield of effective constituent is also undesirable.Chinese patent application (200510042629.3) has reported that a kind of complex enzyme hydrolysis extracts and the production technique of water extraction tea-polyphenol and effective constituents such as by-product trimethyl-xanthine and tea polysaccharide; Chinese patent application (200910040927.7) has been reported effective constituent in a kind of high efficiency extraction Camellia nitidissima Chi leaf and concentrated solution has been applied to technology in the cigarette preparation; Chinese patent application (201210170786.2) has been reported a kind of method that adopts light wave to extract plurality of active ingredients from tealeaves.All do not consider the extraction of same valuable tea albumen in the tea grounds in the above-mentioned patent.
Summary of the invention
The purpose of this invention is to provide a kind of from tealeaves, comprehensively the extraction efficiently and comprise tea-polyphenol, tea polysaccharide, trimethyl-xanthine, theanine, the method for effective constituents such as tea albumen.Present method is raw material with commercially available low and middle-grade green tea, be main extraction solvent with weakly acidic water solution, organic solvent is as the auxiliary extraction solvent, substep extracts tea-polyphenol in the tealeaves, trimethyl-xanthine, tea polysaccharide, theanine, effective constituents such as tea albumen, this method can effectively improve utilization ratio and the productivity effect of tealeaves, reduces preparation cost.Process flow sheet as shown in Figure 1.
The method of effective constituent in a kind of comprehensive extraction tealeaves of the present invention, its step is as follows:
(1) the weakly acidic water solution lixiviate of effective content of tea
Tealeaves is crossed 60～80 mesh sieves, obtain the tea powder, by tea powder: phosphate buffered saline buffer=1:15～1:30(g/mL) solid-to-liquid ratio places phosphate buffered saline buffer with the tea powder, place the water bath with thermostatic control concussion to extract 30～60min this system, repeat to extract, filter and collect tea grounds, merging filtrate namely gets effective content of tea water extract;
(2) extraction of tea albumen
With gained tea grounds oven dry in the step (1), solid-to-liquid ratio (g/mL) by tea grounds: alkali lye=1:20～1:40 places the alkaline NaOH aqueous solution with tea grounds, place the water bath with thermostatic control concussion to extract 60～120min this system, filter cleaner gets the alkaline water extract of tea albumen; Add hydrochloric acid in the alkaline water extract of gained tea albumen, regulate pH and be 4.5 and precipitate, it is tea albumen crude product more than 50% that the precipitation freeze-drying gets purity;
(3) extracting and separating of trimethyl-xanthine
With gained effective content of tea water extract in the step (1), revolve inspissation and be reduced to 1/3 of original volume, make solid content reach 1%～5%, add and the isopyknic chloroform of concentrated solution or methylene dichloride; Concussion extraction trimethyl-xanthine under room temperature, fully after the extraction, the upper strata water separates with lower floor's organic phase, takes off a layer organic phase, and reduction vaporization reclaims organic solvent, and the gained solid substance is multiple water-soluble, and freeze-drying gets purity 85% above trimethyl-xanthine;
(4) extracting and separating of tea-polyphenol
Extraction back gained water in the step (3) is shaken extraction with isopyknic ethyl acetate with it under room temperature, fully after the extraction, upper organic phase is separated with lower layer of water, get upper organic phase, reduction vaporization reclaims ethyl acetate, the gained solid substance is multiple water-soluble, and freeze-drying gets the tea-polyphenol of purity more than 80%;
(5) precipitate and separate of tea polysaccharide and theanine
With extraction back gained water in the step (4), use isopyknic 95% ethanol sedimentation with it, centrifugal supernatant liquor and the precipitation of collecting respectively, the precipitation freeze-drying gets total sugar content 60% above tea polysaccharide; With the freeze-drying of gained supernatant liquor, solid substance dissolves with less water, uses dehydrated alcohol recrystallization repeatedly under low temperature then, gets the theanine of purity more than 65%.
Phosphate buffered saline buffer pH is 5～6 in the described step (1), and the water bath with thermostatic control temperature is 50 ° of C～80 ° C.
Repeating extraction time in the described step (1) is 1～3 time, filters and the collection tea grounds, and merging filtrate obtains effective content of tea water extract.
Described step (2) neutral and alkali NaOH concentration of aqueous solution is 0.06～0.10mol/L, and bath temperature is 70 ° of C～90 ° C, and HCl concentration is 0.05mol/L～0.10mol/L.
Behind extraction time 0.5h～1h that fully condition of extraction is in described step (3) or (4), leave standstill the abundant extraction time.
The ethanol sedimentation number of times is 2 times in the described step (5), and low temperature is-10 ° of C～-20 ° C.
The present invention has the following advantages:
(1) adopt distilled water to extract as primary solvent, with low cost, environmental protection; Extract as secondary solvent with certain amount of organic solvent, avoid the too high and product contamination problem of with an organic solvent bringing in a large number of cost;
(2) adopt step-by-step extracting method, at first extract with weakly acidic water solution, the tea-polyphenol extraction yield improves nearly 30% than the extraction with aqueous solution speed under the same terms, too much tea albumen is conducive to obtain the higher tea-polyphenol of purity by the shortcoming that lixiviate goes out when having avoided weak alkaline aqueous solution to extract; Adopt alkali to put forward the heavy method of acid afterwards and extract tea albumen, method is simple and direct, and cost is low, and can obtain the higher tea albumen of purity;
(3) extract tea-polyphenol in tradition, trimethyl-xanthine on the basis of tea polysaccharide and theanine, has increased the extraction of tea albumen in the tea grounds, has improved utilization ratio and the productivity effect of tealeaves.
(4) method that adopts the washing of repeatedly solvent extraction and deionized water to combine in the leaching process makes effective constituent, extraction solvent and all recyclable utilizations of deionized water that each step uses, reduced production cost effectively, the environmental protection of whole production flow high efficiency, cost are lower, have good using value.
The present invention is main extraction solvent with weakly acidic water solution, and certain amount of organic solvent is extracted tea-polyphenol, trimethyl-xanthine, tea polysaccharide, 4 kinds of effective constituents of theanine as the auxiliary extraction solvent step by step to the water extract; Adopt alkali to put forward the heavy method of acid simultaneously and extract tea albumen in the tea grounds.Advantage of the present invention: (1) is primary solvent with water, and cost is low, and mild condition, is beneficial to the preservation activeconstituents; Extract as secondary solvent with certain amount of organic solvent, avoid the too high and product contamination problem of with an organic solvent bringing in a large number of cost; (2) extract tea-polyphenol, tea polysaccharide, trimethyl-xanthine, theanine and 5 kinds of effective constituents of tea albumen, extraction yield height, purity height, the utilization ratio height of tealeaves simultaneously; (3) all recyclable utilizations of the solvent in the leaching process reduce preparation cost; (4) there are not alkalescence and acid waste liquid discharge, environmental friendliness in a whole set of technology.A complete set of preparation flow is efficient, low-cost, has good using value.
Description of drawings
Fig. 1 integrated extraction technique schema;
The color atlas of tea-polyphenol behind Fig. 2 embodiment 1 purifying.
Embodiment
Embodiment 1:
(1) the weakly acidic water solution lixiviate of effective content of tea
With tealeaves dedusting, the pulverizing of oven dry, cross 60～80 mesh sieves.Get 10g60～80 purpose tea powder, adding 150mL pH is 5 phosphate buffered saline buffer, and then this system being placed temperature is the water bath with thermostatic control concussion extraction of 50 ° of C, behind the extraction 30min, filter, and collection tea grounds drying, it is standby to get supernatant liquor.
(2) extraction of tea albumen
Get the tea grounds 5g that collects in (1) step, adding 100mL concentration by the solid-to-liquid ratio of 1:20 is in the alkaline NaOH aqueous solution of 0.06mol/L, and it is that 60min is extracted in the water bath with thermostatic control concussion of 70 ° of C that this system is placed temperature, filter the alkaline water extract of tea albumen; Add 0.05mol/L hydrochloric acid in the alkaline water extract of tea albumen, regulate pH and be 4.5 and precipitate, the centrifugal supernatant liquor of removing, collecting precipitation, freeze-drying gets tea albumen crude product 816.28mg, and extraction yield is 8.16%, purity 50.59%.
(3) extracting and separating of trimethyl-xanthine
Gained supernatant liquor in (1) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, add and the isopyknic chloroform of concentrated solution or methylene dichloride, fully shake extraction 0.5h under the room temperature, leave standstill 0.5h, collect upper strata water and lower floor's organic phase respectively, organic phase is again with isopyknic chloroform or methylene dichloride room temperature concussion extraction 0.5h, leave standstill 0.5h, collect upper strata water and lower floor's organic phase again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed chloroform or methylene dichloride reclaim standbyly, the solid substance that obtains adds 5mL distilled water washing twice, and lyophilize gets the 10.52mg trimethyl-xanthine, and extraction yield is 1.05%, and purity is 85.5%.
(4) extracting and separating of tea-polyphenol
Merging gained water in (3) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, use isopyknic ethyl acetate, fully concussion extraction 0.5h leaves standstill 0.5h, collects upper organic phase and lower floor's water respectively, organic phase is again with isopyknic ethyl acetate concussion extraction 0.5h, leave standstill 0.5h, collect upper organic phase and lower floor's water again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed ethyl acetate reclaim standbyly, the solid substance that obtains adds 5mL distilled water washing twice, and lyophilize gets the 855.06mg tea-polyphenol, extraction yield 41.1%, and purity is 81.5%.
The color atlas of tea-polyphenol is seen Fig. 2 behind the purifying, wherein contain gallic acid (GA) 5.11%, epi-nutgall acid catechin (EGC) 5.05%, l-Epicatechol (EC) 6.27%, NVP-XAA 723 (EGCG) 50.12%, nutgall catechin gallic acid ester (GCG) 1.67%, L-Epicatechin gallate (ECG) 13.28%.
(5) precipitate and separate of tea polysaccharide and theanine
Revolve inspissation and be reduced to 1/3 of original volume merging the gained water in (4) step, make solid content reach 1%～5%, with isopyknic 95% ethanol sedimentation 2 times, centrifugal supernatant liquor and the precipitation of collecting respectively.The supernatant liquor freeze-drying, solid substance dissolves with less water, uses dehydrated alcohol in-10 ° of C recrystallizations 2 times then, gets the 10.9mg theanine, extraction yield 1.09%, purity 65.2%; Precipitation washes twice with distilled water, and lyophilize gets the 786mg tea polysaccharide, and total sugar content 65.5%, extraction yield are 7.86%.
Embodiment 2:
(1) the weakly acidic water solution lixiviate of effective content of tea
With tealeaves dedusting, the pulverizing of oven dry, cross 60～80 mesh sieves.Get 10g60～80 purpose tea powder, adding 250mL pH is 5～6 phosphate buffered saline buffer, then this system being placed temperature is the water bath with thermostatic control concussion extraction of 70 ° of C, after extracting 40min, filtering tea grounds, to add 250mL pH by above-mentioned steps be that 5.5 phosphate buffered saline buffer extracts 1 time again, filter and collect the tea grounds drying, the supernatant liquor that merges extracted twice is standby.
(2) extraction of tea albumen
Get the tea grounds 5g that collects in (1) step, adding 150mL concentration by the solid-to-liquid ratio of 1:30 is in the alkaline NaOH aqueous solution of 0.08mol/L, and it is that 90min is extracted in the water bath with thermostatic control concussion of 80 ° of C that this system is placed temperature, filter the alkaline water extract of tea albumen; Add 0.075mol/L hydrochloric acid in the alkaline water extract of tea albumen, regulate pH and be 4.5 and precipitate, the centrifugal supernatant liquor of removing, collecting precipitation, freeze-drying gets tea albumen crude product 942.5mg, and extraction yield is 9.43%, and purity is 52.32%.
(3) extracting and separating of trimethyl-xanthine
Gained supernatant liquor in (1) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, with isopyknic chloroform or methylene dichloride, fully concussion extraction 45min leaves standstill 45min, collects upper strata water and lower floor's organic phase respectively, organic phase is again with isopyknic chloroform or methylene dichloride concussion extraction 45min, leave standstill 45min, collect upper strata water and lower floor's organic phase again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed chloroform/methylene dichloride reclaim standbyly, the solid substance that obtains adds 5mL distilled water washing twice, and lyophilize gets the 11.8mg trimethyl-xanthine, and extraction yield is 1.18%, and purity is 87%.
(4) extracting and separating of tea-polyphenol
Merging gained water in (3) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, use isopyknic ethyl acetate, fully concussion extraction 45min leaves standstill 45min, collects upper organic phase and lower floor's water respectively, organic phase is again with isopyknic ethyl acetate concussion extraction 45min, leave standstill 45min, collect upper organic phase and lower floor's water again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed ethyl acetate reclaim standbyly, the solid substance that obtains adds 5mL distilled water washing twice, and lyophilize gets the 1530.2mg tea-polyphenol, extraction yield 78.16%, and purity is 82.7%.
(5) precipitate and separate of tea polysaccharide and theanine
Be concentrated into 1/3 of original volume with merging the gained water in (4) step, make solid content reach 1%～5%, with isopyknic 95% ethanol sedimentation 2 times, centrifugal supernatant liquor and the precipitation of collecting respectively.The supernatant liquor freeze-drying, solid substance dissolves with less water, uses dehydrated alcohol in-15 ° of C recrystallizations 2 times then, gets the 13.9mg theanine, and extraction yield is 1.39%, and purity is 66.4%.Precipitation washes twice with distilled water, and lyophilize gets the 1346.5mg tea polysaccharide, and total sugar content 66.8%, extraction yield are 13.47%.
Embodiment 3:
(1) the weakly acidic water solution lixiviate of effective content of tea
With tealeaves dedusting, the pulverizing of oven dry, cross 60～80 mesh sieves.Get 10g60～80 purpose tea powder, adding 300mL pH is 5～6 phosphate buffered saline buffer, then this system being placed temperature is the water bath with thermostatic control concussion extraction of 80 ° of C, after extracting 60min, filter collecting tea grounds, to add 300mL pH be that 6 phosphate buffered saline buffer extracts 2 times more repeatedly, filter, collect the tea grounds drying, the supernatant liquor that merges 3 extractions is standby.
(2) extraction of tea albumen
Get the tea grounds 5g that collects in (1) step, adding 200mL concentration by the solid-to-liquid ratio of 1:40 is in the alkaline NaOH aqueous solution of 0.10mol/L, and it is that 120min is extracted in the water bath with thermostatic control concussion of 90 ° of C that this system is placed temperature, filter the alkaline water extract of tea albumen; In the alkaline water extract of tea albumen, add 0.10mol/L hydrochloric acid and regulate pH and 4.5 precipitate, the centrifugal supernatant liquor of removing, collecting precipitation, freeze-drying gets tea albumen crude product 1025.8mg, and extraction yield is 10.26%, and purity is 53.56%.
(3) extracting and separating of trimethyl-xanthine
Gained supernatant liquor in (1) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, with isopyknic chloroform or methylene dichloride, fully concussion extraction 1h leaves standstill 1h, collects upper strata water and lower floor's organic phase respectively, organic phase is again with isopyknic chloroform/methylene dichloride concussion extraction 1h, leave standstill 1h, collect upper strata water and lower floor's organic phase again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed chloroform or methylene dichloride reclaim standbyly, the solid substance that obtains adds 5mL distilled water washing twice, and lyophilize gets the 13.6mg trimethyl-xanthine, and extraction yield is 1.36%, and purity is 90%.
(4) extracting and separating of tea-polyphenol
Be concentrated into 1/3 of original volume with merging the gained water in (3) step, make solid content reach 1%～5%, use isopyknic ethyl acetate, fully concussion extraction 1h leaves standstill 1h, collects upper organic phase and lower floor's water respectively, organic phase is again with equal volume of ethyl acetate concussion extraction 1h, leave standstill 1h, collect upper organic phase and lower floor's water again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed ethyl acetate reclaim standbyly, the solid substance that obtains adds 5mL distilled water washing twice, and lyophilize gets the 1838.9mg tea-polyphenol, extraction yield 90.1%, and purity is 83.5%.
(5) precipitate and separate of tea polysaccharide and theanine
Revolve inspissation and be reduced to 1/3 of original volume merging the gained water in (4) step, make solid content reach 1%～5%, with isopyknic 95% ethanol sedimentation 2 times, centrifugal supernatant liquor and the precipitation of collecting respectively.The supernatant liquor freeze-drying, solid substance dissolves with less water, uses dehydrated alcohol in-20 ° of C recrystallizations 2 times then, gets the 14.2mg theanine, and extraction yield is 1.42%, purity 67.1%.Precipitation washes twice with distilled water, and lyophilize gets the 1446.2mg tea polysaccharide, and total sugar content 68.5%, extraction yield are 14.46%.
Embodiment 4:
(1) the weakly acidic water solution lixiviate of effective content of tea
With tealeaves dedusting, the pulverizing of oven dry, cross 60～80 mesh sieves.Get 1kg60～80 purpose tea powder, adding 25L pH is 5.5 phosphate buffered saline buffer, then this system being placed temperature is the water bath with thermostatic control concussion extraction of 70 ° of C, after extracting 40min, filter collecting tea grounds, to add 25L pH be that 5.5 phosphate buffered saline buffer extracts 1 time again, filter and collect the tea grounds drying, the supernatant liquor that merges 2 extractions is standby.
(2) extraction of tea albumen
Get the tea grounds 500g that collects in (1) step, adding 20L concentration by solid-to-liquid ratio 1:40 is in the alkaline NaOH aqueous solution of 0.10mol/L, and it is that 120min is extracted in the water bath with thermostatic control concussion of 90 ° of C that this system is placed temperature, filter the alkaline water extract of tea albumen; In the alkaline water extract of tea albumen, add 0.10mol/L hydrochloric acid and regulate pH and 4.5 precipitate, the centrifugal supernatant liquor that goes, collecting precipitation, freeze-drying gets tea albumen crude product 9.26g, and extraction yield is 9.26%, purity 50.28%.
(3) extracting and separating of trimethyl-xanthine
The supernatant liquor of gained in (1) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, with isopyknic chloroform or methylene dichloride, fully concussion extraction 1h leaves standstill 1h, collects upper strata water and lower floor's organic phase respectively, organic phase is again with isopyknic chloroform or methylene dichloride concussion extraction 1h, leave standstill 1h, collect upper strata water and lower floor's organic phase again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed chloroform/methylene dichloride reclaim standbyly, the solid substance that obtains adds 500mL distilled water washing twice, and lyophilize gets the 1.32g trimethyl-xanthine, and extraction yield is 1.32%, and purity is 90%.
(4) extracting and separating of tea-polyphenol
Merging gained water in (3) step is revolved inspissation be reduced to 1/3 of original volume, make solid content reach 1%～5%, use isopyknic ethyl acetate, fully concussion extraction 1h leaves standstill 1h, collects upper organic phase and lower floor's water respectively, organic phase is again with isopyknic ethyl acetate concussion extraction 1h, leave standstill 1h, collect upper organic phase and lower floor's water again, merge water and organic phase that twice extraction obtains respectively.Organic phase reduction vaporization after merging is removed ethyl acetate reclaim standbyly, the solid substance that obtains adds 500mL distilled water washing twice, and lyophilize gets the 12.39g tea-polyphenol, extraction yield 60.1%, and purity is 81.52%.
(5) precipitate and separate of tea polysaccharide and theanine
Revolve inspissation and be reduced to 1/3 of original volume merging the gained water in (4) step, make solid content reach 1%～5%, with isopyknic 95% ethanol sedimentation 2 times, centrifugal supernatant liquor and the precipitation of collecting respectively.The supernatant liquor freeze-drying, solid substance dissolves with less water, uses dehydrated alcohol in-20 ° of C recrystallizations 2 times then, gets the 1.41g theanine, and extraction yield is 1.41%, purity 65.8%.Precipitation washes twice with distilled water, and lyophilize gets the 14.46g tea polysaccharide, and extraction yield is 14.46%, total sugar content 66.3%.
The method of effective constituent in a kind of comprehensive extraction tealeaves that the present invention proposes, be described by preferred embodiment, person skilled obviously can be changed method as herein described in not breaking away from content of the present invention, spirit and scope or suitably be changed with combination and realize the technology of the present invention.Special needs to be pointed out is, the replacement that all are similar and change apparent to those skilled in the artly, they are regarded as being included in spirit of the present invention, scope and the content.
Claims (6)
1. comprehensive method of extracting effective constituent in the tealeaves is characterized in that step is as follows:
(1) the weakly acidic water solution lixiviate of effective content of tea
Tealeaves is crossed 60～80 mesh sieves, obtain the tea powder, by tea powder: phosphate buffered saline buffer=1:15～1:30(g/mL) solid-to-liquid ratio places phosphate buffered saline buffer with the tea powder, place the water bath with thermostatic control concussion to extract 30～60min this system, repeat to extract, filter and collect tea grounds, merging filtrate namely gets effective content of tea water extract;
(2) extraction of tea albumen
With gained tea grounds oven dry in the step (1), solid-to-liquid ratio (g/mL) by tea grounds: alkali lye=1:20～1:40 places the alkaline NaOH aqueous solution with tea grounds, place the water bath with thermostatic control concussion to extract 60～120min this system, filter cleaner gets the alkaline water extract of tea albumen; Add hydrochloric acid in the alkaline water extract of gained tea albumen, regulate pH and be 4.5 and precipitate, it is tea albumen crude product more than 50% that the precipitation freeze-drying gets purity;
(3) extracting and separating of trimethyl-xanthine
With gained effective content of tea water extract in the step (1), revolve inspissation and be reduced to 1/3 of original volume, make solid content reach 1%～5%, add and the isopyknic chloroform of concentrated solution or methylene dichloride; Concussion extraction trimethyl-xanthine under room temperature, fully after the extraction, the upper strata water separates with lower floor's organic phase, takes off a layer organic phase, and reduction vaporization reclaims organic solvent, and the gained solid substance is multiple water-soluble, and freeze-drying gets purity 85% above trimethyl-xanthine;
(4) extracting and separating of tea-polyphenol
Extraction back gained water in the step (3) is shaken extraction with isopyknic ethyl acetate with it under room temperature, fully after the extraction, upper organic phase is separated with lower layer of water, get upper organic phase, reduction vaporization reclaims ethyl acetate, the gained solid substance is multiple water-soluble, and freeze-drying gets the tea-polyphenol of purity more than 80%;
(5) precipitate and separate of tea polysaccharide and theanine
With extraction back gained water in the step (4), use isopyknic 95% ethanol sedimentation with it, centrifugal supernatant liquor and the precipitation of collecting respectively, the precipitation freeze-drying gets total sugar content 60% above tea polysaccharide; With the freeze-drying of gained supernatant liquor, solid substance dissolves with less water, uses dehydrated alcohol recrystallization repeatedly under low temperature then, gets the theanine of purity more than 65%.
2. the method for claim 1 is characterized in that phosphate buffered saline buffer pH is 5～6 in the step (1), and the water bath with thermostatic control temperature is 50 ° of C～80 ° C.
3. the method for claim 1 is characterized in that repeating extraction time according to step (1) is 1～3 time, filters and the collection tea grounds, and merging filtrate obtains effective content of tea water extract.
4. the method for claim 1 is characterized in that step (2) neutral and alkali NaOH concentration of aqueous solution is 0.06～0.10mol/L, and bath temperature is 70 ° of C～90 ° C, and HCl concentration is 0.05mol/L～0.10mol/L.
5. the method for claim 1, it is characterized in that fully the condition of extraction is extraction time 0.5h～1h in described step (3) or (4) after, leave standstill the abundant extraction time.
6. the method for claim 1 is characterized in that the ethanol sedimentation number of times is 2 times in the step (5), and low temperature is-10 ° of C～-20 ° C.
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| CN103891964A (en) * | 2014-04-20 | 2014-07-02 | 张文艳 | Method for massively extracting caffeine from tea |
| CN104829486A (en) * | 2015-06-04 | 2015-08-12 | 成都华高生物制品有限公司 | Method for extracting high-purity theanine from fresh tea leaves |
| CN105713067A (en) * | 2014-08-13 | 2016-06-29 | 肖斌 | Industrial extraction method of selenium protein from selenium-enriched tea |
| CN105712897A (en) * | 2015-01-25 | 2016-06-29 | 宜昌绿源生物技术有限公司 | Tea polyphenol extracting and recycling method |
| CN105859801A (en) * | 2016-04-05 | 2016-08-17 | 林云权 | Oolong tea polyphenol and extraction method thereof |
| CN106905412A (en) * | 2017-02-09 | 2017-06-30 | 上海师范大学 | Tea polysaccharide, the method for glycoprotein in a kind of removal Se rich tea albumen |
| CN107759599A (en) * | 2017-10-12 | 2018-03-06 | 贵州省石阡正岩苔生态茶业有限公司 | A kind of method of tealeaves extraction caffeine inferior |
| CN109497218A (en) * | 2019-01-15 | 2019-03-22 | 福建仙洋洋生物科技有限公司 | A kind of technique of production energy-saving high-quality tea concentrate |
| CN109535270A (en) * | 2018-10-31 | 2019-03-29 | 浙江医院 | A kind of preparation method of Anji white tea polysaccharide and its new application of anti-human cytomegalovirus |
| CN109601673A (en) * | 2018-12-19 | 2019-04-12 | 统企业(中国)投资有限公司昆山研究开发中心 | Clarification type ground coffee and preparation method thereof |
| CN110272354A (en) * | 2019-03-08 | 2019-09-24 | 贺州学院 | The technique of theanine is extracted from tea polyphenols waste liquid using foam separating technology |
| CN111358045A (en) * | 2020-03-13 | 2020-07-03 | 颐中（青岛）实业有限公司 | Black tea extract compound and application thereof |
| CN112858604A (en) * | 2021-01-06 | 2021-05-28 | 华润怡宝饮料（中国）有限公司 | Evaluation method of tea extraction end point |
| CN113190066A (en) * | 2021-04-29 | 2021-07-30 | 佛山市龙生光启科技有限公司 | High-purity EGCG automatic extraction system based on Internet of things |
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| CN103891964A (en) * | 2014-04-20 | 2014-07-02 | 张文艳 | Method for massively extracting caffeine from tea |
| CN105713067A (en) * | 2014-08-13 | 2016-06-29 | 肖斌 | Industrial extraction method of selenium protein from selenium-enriched tea |
| CN105712897A (en) * | 2015-01-25 | 2016-06-29 | 宜昌绿源生物技术有限公司 | Tea polyphenol extracting and recycling method |
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| CN105859801A (en) * | 2016-04-05 | 2016-08-17 | 林云权 | Oolong tea polyphenol and extraction method thereof |
| CN105859801B (en) * | 2016-04-05 | 2017-12-05 | 林云权 | A kind of oolong tea polyphenol and its extracting method |
| CN106905412A (en) * | 2017-02-09 | 2017-06-30 | 上海师范大学 | Tea polysaccharide, the method for glycoprotein in a kind of removal Se rich tea albumen |
| CN106905412B (en) * | 2017-02-09 | 2020-08-07 | 上海师范大学 | Method for removing tea polysaccharide and glycoprotein in selenium-rich tea protein |
| CN107759599A (en) * | 2017-10-12 | 2018-03-06 | 贵州省石阡正岩苔生态茶业有限公司 | A kind of method of tealeaves extraction caffeine inferior |
| CN109535270A (en) * | 2018-10-31 | 2019-03-29 | 浙江医院 | A kind of preparation method of Anji white tea polysaccharide and its new application of anti-human cytomegalovirus |
| CN109601673A (en) * | 2018-12-19 | 2019-04-12 | 统企业(中国)投资有限公司昆山研究开发中心 | Clarification type ground coffee and preparation method thereof |
| CN109601673B (en) * | 2018-12-19 | 2021-09-17 | 统一企业(中国)投资有限公司昆山研究开发中心 | Clear coffee powder and preparation method thereof |
| CN109497218A (en) * | 2019-01-15 | 2019-03-22 | 福建仙洋洋生物科技有限公司 | A kind of technique of production energy-saving high-quality tea concentrate |
| CN110272354A (en) * | 2019-03-08 | 2019-09-24 | 贺州学院 | The technique of theanine is extracted from tea polyphenols waste liquid using foam separating technology |
| CN111358045A (en) * | 2020-03-13 | 2020-07-03 | 颐中（青岛）实业有限公司 | Black tea extract compound and application thereof |
| CN112858604A (en) * | 2021-01-06 | 2021-05-28 | 华润怡宝饮料（中国）有限公司 | Evaluation method of tea extraction end point |
| CN113190066A (en) * | 2021-04-29 | 2021-07-30 | 佛山市龙生光启科技有限公司 | High-purity EGCG automatic extraction system based on Internet of things |
| CN113190066B (en) * | 2021-04-29 | 2022-04-01 | 广西河丰药业有限责任公司 | High-purity EGCG automatic extraction system based on Internet of things |
| CN116235894A (en) * | 2023-03-02 | 2023-06-09 | 大闽食品（漳州）有限公司 | Preparation method and application of tea dreg protein |
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