CN1301698A - Method for separating catechin monomer from tea extration - Google Patents
Method for separating catechin monomer from tea extration Download PDFInfo
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- CN1301698A CN1301698A CN 99127607 CN99127607A CN1301698A CN 1301698 A CN1301698 A CN 1301698A CN 99127607 CN99127607 CN 99127607 CN 99127607 A CN99127607 A CN 99127607A CN 1301698 A CN1301698 A CN 1301698A
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Abstract
In separation of catechin monometer from the tea extract, a countercurrent chromatograph is used as separating equipment and a solvent system comprising alkane, ethyl acetate and water with or without further addition of alcohol and acetic acid, and they being liquids at normal temperature and normal pressure is used. The separation is an economic process utilizing simple equipment and cheap solvent system.
Description
The present invention relates to deep processing field of tea, particularly a kind of from tea extract the separating catechin monomer methods.
Tea extract is the mixing active ingredients thing that extracts from tealeaves, and catechin is a wherein topmost active component, again with EGCG, ECG, the tool economic worth of GCG, EGC, has been listed in a kind of potential cancer therapy drug in the catechin.The method of the above-mentioned catechin monomers of separation and Extraction adopted Sephedax-LH20 gel column chromatography (WilkinsC.K.et al from tealeaves in the past, J.Sci.Food Agri., 22,1977,480.) just separate with Sephedax-LH20-preparative high performance liquid chromatography separates (Yuyabe F.Etal again, Nippon NogeikagakuKaishi, 1989,63 (4): 845).Its limitation is that the Sephedax-LH20 gel costs an arm and a leg, and difficulty is used in regeneration, thereby the monomer production cost is higher; High performance liquid chromatography is chromatographic column costliness not only then, and equipment is also complicated, is a kind of more uneconomic method.
The object of the present invention is to provide a kind of separating catechin monomer methods from tea extract of economy.
For achieving the above object, the present invention is by the following technical solutions: it with counter current chromatograph as separating device; Its solvent systems is by being in liquid alkane under the normal temperature and pressure, ethyl acetate and water are formed, the volumetric ratio of alkane and ethyl acetate is 1: 4～12, the consumption of water should guarantee to make phase layering up and down at least, perhaps described solvent systems is by being in liquid alkane under the normal temperature and pressure, ethyl acetate, alcohol, water and acetate are formed, the volumetric ratio of alkane and ethyl acetate is 1: 1～4, ethyl acetate is 1: 1～3 with the volumetric ratio of alcohol, the volumetric ratio of acetate and alcohol is 1: 4～50, the consumption of water should guarantee to make phase layering up and down at least, above-mentioned alkane is the mixture of single kind of alkane or several alkane, and above-mentioned alcohol is propyl alcohol, butanols, hexanol or enanthol;
And carry out according to the following steps successively:
1). the configuration solvent systems, treat static layering after, upper and lower two-phase is separately standby, will inject the chromatographic column of counter current chromatograph on the above-mentioned solvent systems mutually, get phased soln tea extract under the part of above-mentioned solvent systems;
2). open counter current chromatograph, inject above-mentioned dissolved tea extract and remaining phase down; Its injection order can be to inject above-mentioned dissolved tea extract earlier to inject remaining phase down then, also can be inject earlier residue down the part of phase inject remaining following phase then with the pre-equilibration above-mentioned dissolved tea extract that reinjects, also can be to inject the remaining above-mentioned dissolved tea extract that injects then mutually down earlier;
3). regularly collect elutriant.
When adopting above technical scheme, the present invention also can adopt following further technical scheme:
Described alkane is best with the sherwood oil, and described alcohol is best with the propyl carbinol.
Owing to adopted technique scheme, counter current chromatograph is a General Instrument, its more efficient liquid chromatograph far is a simple cheap, its solvent systems is prepared by usual vehicle, price is also cheap far beyond chromatographic column of the prior art, is a kind of separating catechin monomer methods from tea extract of economy therefore.
The synoptic diagram of the 10L-V high-speed counter-current chromatograph that Fig. 1 is adopted for embodiment 5,6,7.
The invention will be further described below in conjunction with drawings and Examples.
Embodiment 1
The solvent systems of present embodiment adopts sherwood oil: ethyl acetate: water=1: 8: 20, counter current chromatograph adopts homemade GS-10A high-speed counter-current chromatograph, the consisting of of tea extract: EGCG accounts for 23.80%, ECG accounts for 7.0%, GCG accounts for 14.80%, EC accounts for 7.22%, EGC accounts for 10.90%, DL-C accounts for 4.6%, caffeine accounts for 7.57%, all the other 24% are not clear thing.
Get sherwood oil 30ml, ethyl acetate 240ml, water 600ml, mix jolting 20 times in the 1000ml separating funnel, behind the standing demix, two-phase is respectively charged into reagent bottle up and down.Last flow velocity with 10ml/min is pumped into the adverse current chromatogram post.Take by weighing tea extract 3 and restrain in the 50ml reagent bottle, with phase solution dissolving under the 40ml.Open counter current chromatograph to 800 commentaries on classics/min, the flow velocity with 1ml/min pumps into tea extract solution then, uses the following to the catechin composition in the 1ml/min flow velocity wash-out post of remainder instead after treating into to finish.Press 5ml/ pipe or 10ml/ pipe collection elutriant with Fraction Collector, the elution time section of every kind of catechin monomers can be determined by monitor when preparing for the first time.Collect the dry catechin monomers 2090mg of getting of each one-component, wherein EGCG720mg purity 81%, ECG220mg purity 87%, GCG account for 450mg purity 90%, EGC350mg purity 75%, EC+DL-C350mg.
The solvent systems of present embodiment adopts pentane: ethyl acetate: water=1: 5: 10, counter current chromatograph adopts homemade GS-10A high-speed counter-current chromatograph, the consisting of of tea extract: EGCG accounts for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get pentane 60ml, ethyl acetate 300ml, water 600ml, mix jolting 20 times in the 1000ml separating funnel, behind the standing demix, two-phase is respectively charged into reagent bottle up and down.Last flow velocity with 10ml/min is pumped into the adverse current chromatogram post.Take by weighing tea extract 1.5 and restrain in the 50ml reagent bottle, with phase solution dissolving under the 40ml.Open counter current chromatograph to 600 commentaries on classics/min, flow velocity with 1.4ml/min pumps into down 30 minutes mutually then, again tea extract solution is pumped into chromatographic column with the flow velocity of 1.4ml/min, treat to use instead after into intact remaining following with the catechin composition in the 1.4ml/min flow velocity wash-out post.Press 5ml/ pipe or the 10ml/ pipe is collected elutriant with Fraction Collector, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG800mg purity 87% that gets.
The solvent systems of present embodiment adopts heptane: ethyl acetate: water=1: 11: 40, counter current chromatograph adopts homemade GS-10A high-speed counter-current chromatograph, the consisting of of tea extract: EGCG accounts for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get heptane 25ml, ethyl acetate 275ml, water 1000ml, mix jolting 20 times in the 1500ml separating funnel, behind the standing demix, two-phase is respectively charged into reagent bottle up and down.Last flow velocity with 10ml/min is pumped into the adverse current chromatogram post.Take by weighing tea extract 3 and restrain in the 50ml reagent bottle, with phase solution dissolving under the 100ml.Open counter current chromatograph to 900 commentaries on classics/min, the flow velocity with 1.4ml/min pumps into tea extract solution then, uses the following to the catechin composition in the 1.4ml/min flow velocity wash-out post of remainder instead after treating into to finish.Press 5ml/ pipe or the 10ml/ pipe is collected elutriant with Fraction Collector, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG1700mg purity 87% that gets.
Embodiment 4
The solvent systems of present embodiment adopts sherwood oil: ethyl acetate: propyl carbinol: water: acetate=1: 1.5: 3: 12: 0.60, counter current chromatograph adopts homemade GS-10A high-speed counter-current chromatograph, the consisting of of tea extract: EGCG accounts for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get sherwood oil 40ml, ethyl acetate 60ml, propyl carbinol 120ml, water 480ml, acetate 24ml, mix jolting 20 times in the 1000ml separating funnel, behind the standing demix, two-phase is respectively charged into reagent bottle up and down.Last flow velocity with 10ml/min is pumped into the adverse current chromatogram post.Take by weighing tea extract 1.5 and restrain in the 50ml reagent bottle, with phase solution dissolving under the 12ml.Open counter current chromatograph to 400 commentaries on classics/min, the flow velocity with 2ml/min pumps into tea extract solution then, uses the following to the catechin composition in the 2ml/min flow velocity wash-out post of remainder instead after treating into to finish.Press 5ml/ pipe or the 10ml/ pipe is collected elutriant with Fraction Collector, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG780mg purity 87% that gets.
The counter current chromatograph of present embodiment adopts the 10L-V high-speed counter-current chromatograph, and its structure is as follows:
With reference to accompanying drawing 1.It comprises housing 1, can manage 4,5 around the separator column 3 of its axis 2 rotations and leading in/out of being connected with separator column, the outside of separator column also is provided with the rotatable fixed frame 6 that leads in/out pipe, its turning axle and above-mentioned axis coaxial line, the coupling end that leads in/out pipe and separator column passes axis, and the other end passes the Partner 7 of turning axle.
One end of fixed frame 6 is provided with link span 8, and connects by the power output shaft 10 of link span and motor 9, and the bearing 11 of the Partner 7 of turning axle is located on the housing 1.The bearing 12,13 of axis is located on the fixed frame 6, the power input shaft 14 of separator column passes framework, the coaxial gear 15,16 that is provided with in its two ends, gear 15 is connected by belt or carrier gear 18 with in-line gears 17 on the power output shaft 10, in-line gears 19 toe joints on gear 16 and the axis.The inwall of forming the pipe of separator column is screw-like or corrugated, and is the Teflon pipe.The described pipe that leads in/out is the Teflon pipe.
The solvent systems of present embodiment adopts hexane: ethyl acetate: enanthol: water: acetate=1: 2.5: 7: 25: 0.35, the consisting of of tea extract: EGCG accounted for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get hexane 1600ml, ethyl acetate 2400ml, enanthol 10200ml, water 40000ml, acetate 560ml, mix in 100000ml mixing-layering jar, behind the standing demix, two-phase is separately standby up and down.Last flow velocity with 40ml/min is pumped into or be pressed into the adverse current chromatogram post with the air pressure mode.Take by weighing tea extract 120 grams, with phase solution dissolving under the 2000ml.Open counter current chromatograph to 25 commentariess on classics/min, pump into down with the flow velocity of 5ml/min then that the flow velocity with 30ml/min pumped into tea extract solution again with pre-equilibration in 1 hour mutually, use the following of remainder instead after treating into to finish with the catechin composition in the 5ml/min flow velocity wash-out post.Regularly collect elutriant, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG62g purity 85% that gets.
Embodiment 6
The solvent systems of present embodiment adopts sherwood oil: ethyl acetate: amylalcohol: water: acetate=1: 3.5: 4.2: 35: 0.14, counter current chromatograph adopts the 10L-V high-speed counter-current chromatograph, the consisting of of tea extract: EGCG accounts for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get sherwood oil 2500ml, ethyl acetate 8750ml, amylalcohol 10500ml, water 87500ml, acetate 350ml, mix in 120000ml mixing-layering jar, behind the standing demix, two-phase is separately standby up and down.Last flow velocity with 40ml/min is pumped into or be pressed into the adverse current chromatogram post with the air pressure mode.Take by weighing tea extract 60 grams, with phase solution dissolving under the 1500ml.Open counter current chromatograph to 15 commentaries on classics/min, the flow velocity with 30ml/min pumps into tea extract solution then, uses the following to the catechin composition in the 10ml/min flow velocity wash-out post of remainder instead after treating into to finish.Regularly collect elutriant, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG42g purity 87% that gets.
The solvent systems of present embodiment adopts sherwood oil: ethyl acetate: propyl carbinol: water: acetate=1: 4: 8: 40: 0.2, counter current chromatograph adopts the 10L-V high-speed counter-current chromatograph, the consisting of of tea extract: EGCG accounts for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get sherwood oil 2500ml, ethyl acetate 10000ml, propyl carbinol 20000ml, water 100000ml, acetate 500ml, mix in 150000ml mixing-layering jar, behind the standing demix, two-phase is separately standby up and down.Last flow velocity with 40ml/min is pumped into or be pressed into the adverse current chromatogram post with the air pressure mode.Take by weighing tea extract 180 grams, with phase solution dissolving under the 1500ml.Open counter current chromatograph to 15 commentaries on classics/min, the flow velocity with 30ml/min pumps into tea extract solution then, uses the following to the catechin composition in the 5ml/min flow velocity wash-out post of remainder instead after treating into to finish.Regularly collect elutriant, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG93g purity 82% that gets.
Embodiment 8
The solvent systems of present embodiment adopts sherwood oil: ethyl acetate: propyl carbinol: water: acetate=1: 2: 4: 45: 0.08, counter current chromatograph adopts U.S. CPC-828 orthogonal axis type high-speed counter-current chromatograph (Y.Shibusawa and Y.Ito, J.Chromatogr.596:118～122,1992), consisting of of tea extract: EGCG accounts for 50.80%, ECG accounts for 7.01%, GCG accounts for 1.80%, EC accounts for 1.22%, EGC accounts for 5.90%, DL-C accounts for 0.6%, caffeine accounts for 1.57%, all the other are not clear thing.
Get sherwood oil 100ml, ethyl acetate 200ml, propyl carbinol 400ml, water 4500ml, acetate 8ml, mix jolting 20 times in the 6000ml separating funnel, behind the standing demix, two-phase is respectively charged into reagent bottle up and down.Last flow velocity with 20ml/min is pumped into the adverse current chromatogram post.Take by weighing tea extract 10 grams, with phase solution dissolving under the 150ml.Open counter current chromatograph to 500 commentaries on classics/min, the flow velocity with 20ml/min pumps into tea extract solution then, uses the following to the catechin composition in the 3ml/min flow velocity wash-out post of remainder instead after treating into to finish.Regularly collect elutriant, the determining of catechin monomers elution time section with embodiment 1.The dry EGCG5g purity 82% that gets.
Claims (2)
1, a kind of from tea extract the separating catechin monomer methods, it is characterized in that it with counter current chromatograph as separating device; Its solvent systems is by being in liquid alkane under the normal temperature and pressure, ethyl acetate and water are formed, the volumetric ratio of alkane and ethyl acetate is 1: 4～12, the consumption of water should guarantee to make phase layering up and down at least, perhaps described solvent systems is by being in liquid alkane under the normal temperature and pressure, ethyl acetate, alcohol, water and acetate are formed, the volumetric ratio of alkane and ethyl acetate is 1: 1～4, ethyl acetate is 1: 1～3 with the volumetric ratio of alcohol, the volumetric ratio of acetate and alcohol is 1: 4～50, the consumption of water should guarantee to make phase layering up and down at least, above-mentioned alkane is the mixture of single kind of alkane or several alkane, and above-mentioned alcohol is propyl alcohol, butanols, alcohol or enanthol;
And carry out according to the following steps successively:
1). the configuration solvent systems, treat static layering after, upper and lower two-phase is separately standby, will inject the chromatographic column of counter current chromatograph on the above-mentioned solvent systems mutually, get phased soln tea extract under the part of above-mentioned solvent systems;
2). open counter current chromatograph, inject above-mentioned dissolved tea extract and remaining phase down;
3). regularly collect elutriant.
2, as claimed in claim 1 a kind of from tea extract the separating catechin monomer methods, it is characterized in that described alkane is best with the sherwood oil, described alcohol is best with the propyl carbinol.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991276078A CN1159308C (en) | 1999-12-28 | 1999-12-28 | Method for separating catechin monomer from tea extration |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991276078A CN1159308C (en) | 1999-12-28 | 1999-12-28 | Method for separating catechin monomer from tea extration |
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| CN1301698A true CN1301698A (en) | 2001-07-04 |
| CN1159308C CN1159308C (en) | 2004-07-28 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100422188C (en) * | 2006-08-25 | 2008-10-01 | 浙江大学 | Method for separating preparing tripterygium wilfordii monomer from tripterygium wilfordii by countercurrent flow chromatography |
| CN100582102C (en) * | 2007-12-07 | 2010-01-20 | 西南大学 | Process for preparing methylation catechin by tea |
| CN105218503A (en) * | 2014-07-05 | 2016-01-06 | 中华全国供销合作总社杭州茶叶研究所 | Tea catechin preparation technology in tea fresh leaves |
-
1999
- 1999-12-28 CN CNB991276078A patent/CN1159308C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100422188C (en) * | 2006-08-25 | 2008-10-01 | 浙江大学 | Method for separating preparing tripterygium wilfordii monomer from tripterygium wilfordii by countercurrent flow chromatography |
| CN100582102C (en) * | 2007-12-07 | 2010-01-20 | 西南大学 | Process for preparing methylation catechin by tea |
| CN105218503A (en) * | 2014-07-05 | 2016-01-06 | 中华全国供销合作总社杭州茶叶研究所 | Tea catechin preparation technology in tea fresh leaves |
| CN105218503B (en) * | 2014-07-05 | 2018-10-19 | 中华全国供销合作总社杭州茶叶研究所 | Tea catechin preparation process in fresh tea leaves |
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| CN1159308C (en) | 2004-07-28 |
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