JPH0314572A - Production of catechins of plant body - Google Patents

Abstract

PURPOSE:To obtain safe, inexpensive and high-purity catechins having antioxidizing, deodorizing and antimicrobial action by efficiency extracting catechins which are active ingredients from a plant body with a specific solvent and purifying the resultant catechins without using toxic substances. CONSTITUTION:Catechins, such as (+)-catechin, (-)-epicatechin or gallic acid, are extracted from a plant body especially obtained from tea (Camellia sinensis var. sinensis, etc.) as a raw material using an aqueous solution of a lower alcohol and the resultant extract is then adsorbed on gel beads and subsequently washed with water to liberate impurities other than the catechins adsorbed on the gel beads with an aqueous solution of a lower alcohol in a low concentration. The catechins adsorbed on the gel beads are finally isolated with an aqueous solution of a lower alcohol in a high concentration to afford the catechins on a commercial scale. The above-mentioned catechins hardly contain any impurities, such as caffeine, and can be used as a natural food additive or an additive of confectionery for children, tooth powder, etc.

Description

[Detailed description of the invention]

(Industrial field in Icheon) The present invention extracts catechins contained in plants using lower alcohol, especially safe ethanol, and adsorbs Rf+ to a gel material such as gel beads to reduce the concentration. This invention relates to a method for producing catechins, which involves removing impurities and producing catechins using different lower alcohols. (Prior art) As a method for extracting catechin from tea confectionery and producing pods, it is extracted from MM with aqueous acetone, further extracted with ethyl alcohol, and then extracted with Sephadex LH-20YaI-aH/l. , II W-40 column +: adsorption,
The p method for preparing tn is known, but it is used for preparing analytical samples or on a laboratory scale. Moreover, Sephadex L
II-20 and Toyopearl II W-40
fl i! of the sample against the gel! ! This method has a small f m and also serves as a partial production of catechins by the Chroma 1 method using a column, and it is a method for producing large ffi sheaths of catechins contained in Okawa and about 15% of the dry worm dish in a tea towel. is inappropriate. In addition, as a method for extracting ingredients in tea confectionery, the commercial standard IQ is 1 liter of boiled water for producing canned drinks and instant tea.
There is only a method for extracting organic solvents, and there is no method for using organic solvents. Antioxidant effect (Japanese Patent Application Laid-Open No. 60-1450
70) and the harmful effects of N-nitrosamine formation (Special Publication No. 6)
2-11571) has been obtained, but
The content of catechins, which have these effects, is low, and the mixture also contains many impurities such as caffeine. In addition, techniques that can be applied to the i method for removing caffeine contained in extracts include various decaffeination methods from coffee, black tea, etc.; 61-22934), ■Supercritical liquid carbonic acid extraction method (Japanese Patent Publication No. 59-10775, 59-
41377) +3 Fatty {{extraction with material (1986-41378), <3) Hot water +1Q treatment method 11 (1977-41 69 2.), ■Caffeine-tannin complex precipitation Qth (1ff public 11Q6 1-5
1 8 4 9 ), @ 4M B'a Adsorption method (Japanese Patent Publication No. 60-5249), ■ Method of adsorbing activated carbon, silica gel, activated alumina, locust and soybean as suction cuttings (Japanese Patent Publication No. 59-39097, No. 59-46576) ) etc. are known. Among these, from the viewpoint of selectively removing caffeine from a mixture of catechins and caffeine, ■
It is expected that the ester type catechins will be decomposed by heat, and ■
There is a concern that thermal decomposition of gallic acid esters, which are common in tea confectionery, will occur; in ■, a loss of many catechins is expected; and in ■ and ■, non-selective adsorption of not only caffeine but also catechins is expected. On the other hand, methods ① and ③ are considered as methods that may be able to selectively remove only caffeine from the tea extract mixture. However, in the selective removal of caffeine using halogen (chlorine)-containing organic solvents such as chloroform and methylene chloride (Japanese Patent Publication No. 61-229-34), the solvent itself is toxic and carcinogenic. It poses a problem if it remains in food, and its use as a decaf method is prohibited in the United States. In addition, although extraction using liquid carbonic acid in a supercritical state (■) produces safe extracts, it requires large-scale extraction equipment that is subject to regulations under the High Pressure Gas Control Act, and the extraction costs are high, resulting in lower unit prices. There is no practical use for anything other than high *m extraction and bamboo production. (Problem N to be solved by the invention) The purpose of the present invention is to efficiently extract catechins, which are active ingredients, from plants and produce them without using harmful substances, thereby producing a safe and inexpensive solution. , antioxidant effect, /I!
The purpose is to remove catechins, which have odor and antibacterial effects and contain almost no impurities other than the active ingredients. These catechins are fat-soluble and water-soluble, and can be used as additives in natural foods, as well as in confectionery for children, polishing powder, etc., which contain almost no caffeine. (Means for solving the problem) The catechins contained in tea confectionery are known to have antioxidant effects, antibacterial effects, mutation suppressing effects, blood cholesterol regulating effects, angiotensin I exchange enzyme suppressing effects, etc. There is. In particular, it is expected to be used as food additives, such as antioxidants and antibacterial agents extracted from natural products. However, when extracting kachkin from tea confectionery with hot water or a solvent, caffeine is always extracted. There is no commercially available method for safely removing the caffeine contained in the mixture. In addition, as a method for decaffeinating coffee and tea, a method is known in which it is adsorbed onto a nonionic hydrophobic polymer resin (Japanese Patent Publication No. 00-5249). I II<
There is no point in removing it. Therefore, conventional products extracted from catechins by the absorption method using T4 fat are mixtures containing caffeine, and the future of this product has been determined. Therefore, the purpose of the present invention is to provide the following method as an efficient extraction and production method for catechins from plants, which can be added to foods in consideration of safety.In particular, Extracting with alcohol, ■
It is characterized by increasing the content of the extract relative to gel beads and producing a highly pure catechin mixture that contains almost no caffeine. (1) Method for extracting catechins from a hammered object Using an aqueous solution of lower alcohols, especially an aqueous solution of ethanol,
1 ゜▼ar. sls6nglg, Hr. *ssmaica
etc.) and leaves of plants of the genus Camellia containing catechins and their hybrids, other {M-body components, and mallet objects including processed products thereof, especially from manufactured tea and tea leaves, gallic T-monoacid, {■one-catechin, {1)one-shrimp catechin, <<-1-shrimp gallocatechin, (-)mono- and digallate ester of one-shrimp gallocatechin, (
-) Mono- and digallate esters of epicatechin, as well as catechins that are double or triple east combinations or monomers containing these compounds as eight components, are extracted. Lower alcohol aqueous solution is 15 to 95
Body In %, good * L < Ha 5 0 ~ 8 5 bodies? n
% is used. Manufactured tea refers to processed tea such as sencha, steaming process of tea manufacturing process, leaf beating process, rough tea process,
It is also possible to use dried tea after any process such as rolling or medium rolling, oolong tea or black tea that has been subjected to Iff treatment, and powders and tea waste produced during the process of turning these into products. These dried plants have a moisture content of 15%.
The diameter must be 2 mm or less. Raw plant leaves and their components can be extracted by adding 7 g of lower alcohol aqueous solution and crushing them using a mixer or the like. 1 to 100 times as much as Higashim to crushed stem 1 budai or raw plant leaves and their pastes,
Preferably, add a lower alcohol aqueous solution of 5 to 20 times the strength, and heat for 10 minutes to 24 hours, preferably 30 minutes to 14 hours, 10' to 100,011 revolutions/min, or preferably 40 to 400 revolutions/min. Stir with or 3 to 500
Catechins are extracted by mixing by scooping at a rate of 10 to 100 times/minute. The extract obtained by fractionation using a centrifuge or filtration using paper, filters, etc. is preferably subjected to nitrogen purging to reduce the water content by freeze-drying or turning it into a dry powder to prevent desiccation polymerization of catechins. Then, store it in a dark place at low temperature. (2) Adsorption of plant stamina chicken extract onto gel beads Hydrophilic vinyl polymer beads (Toyobar ItW-40
Gel beads such as (+1 made by Tosoh), Dekiss!・Gel beads with acylated or acetylated (hydroxyglopylated) hydroxyl groups of starch gels such as orchids (gel beads such as Sephadex L II-2 0, 60, manufactured by GXPharmacia), agarose-based gels Hyperchroma 1 Graffi River Gel Beads

[Gel beads such as Biogel A (manufactured by Bio-Rad Laboratories)] and gel-like substances, etc. Yes! 11 Place an extract of a lower alcohol aqueous solution of a plant or a dried product thereof onto gel beads resistant to FB medium, and the extract is 1 to 5.
0! Dilute with water to TL 4%, preferably 5 to 25% by weight, and dry in a heavy plate to give 20 to 1000g% OF, preferably 100 to 3%, based on the weight of the gel beads.
00 g of plant extract is loaded and allowed to adsorb for 5 minutes to 24 hours, preferably 30 minutes to 4 hours. The particle size of the gel beads used is 5μIn to 2mm, preferably 50μIn to 300μm. Adsorption, washing, desorption, etc. to gel beads are done using gel beads. Both the batch method, which is carried out in eight chambers, and the column method, in which the solvent passes through the chamber, are both 1iR. (3) Washing of gel beads that have adsorbed 1lh extract Gel beads that have adsorbed plant adsorbates are washed using distilled water, mt ion water, and water with a low mold content of σ and other ions. In the batch method, the gel bead body IH is 175 to 100
Use twice as much water or 1 to 5 times as much water, wash for 5 minutes to 24 hours, preferably 10 minutes to 4 hours, and stir at 10-1000 rpm or 3 times. ~
Perform by shaking 500 times/min. Repeat this operation up to 5 times, preferably 2 to 3 times. In the column method, water is used in an amount of 1 to 1000 times as much water as the gel bead body f7[, and the water is continued to flow until the absorbance of 2801111 of the ultraviolet detector decreases and becomes constant.
One. (4) Release and removal of caffeine and other impurities other than catechins from the gel beads adsorbed with the plant extract using a low concentration lower alcohol aqueous solution.
When brought into contact with a lower alcohol aqueous solution of W degree, caffeine etc. are liberated, and by washing and separating, only catechins can be adsorbed to the gel beads and made to be irrigated. The low concentration aqueous solution used for this operation is 5-4
It is an aqueous solution containing 0% lower alcohol, preferably 15-25% lower alcohol. Note that specific examples of lower alcohols are the same as those mentioned above. In the batch method, a low concentration lower alcohol aqueous solution is used that is 1/5 to 100 times the volume of the gel beads, preferably 1 to 5 times the volume, and the washing time is 5 minutes to 24 hours, preferably 20 minutes to 40 hours. Within 10~1000I! ! 1
Stir at 3 to 500 revolutions per minute or shake at 3 to 500 times per minute. Repeat this operation up to 5 times, preferably 2 to 3 times. In the column method, the gel bead physical size is 1 to 10 o o 1
0. Ultraviolet ray detection of low concentration lower alcohol aqueous solution! Continue to flow until the absorbance at 280 nm of 1 decreases and becomes constant, and increase the yi separation to {7. (5) Releasing and regenerating catechins from gel beads washed with a low concentration lower alcohol aqueous solution using a high concentration lower alcohol aqueous solution Gel beads are washed with a low concentration lower alcohol aqueous solution to release and wash and separate caffeine, etc. When contacted with an appropriately high concentration lower alcohol aqueous solution, desorption of catechins occurs. The highly concentrated aqueous solution used in this operation is an aqueous solution containing lower alcohol in an amount of 40 to 95% by volume, preferably 50 to 85% by volume. Note that specific examples of lower alcohols are the same as those listed in nu. In batch mode, 175-10 of the volume of gel beads
Use a highly concentrated lower alcohol aqueous solution of 0 times the volume, preferably 1 to 5 times the amount, and wash for 5 minutes to 24 hours, preferably 20 minutes to 4 hours, stirring at 10 to 1000 concave rotations/min. It is carried out with a tithe or with a raw iron at a rate of 3 to 500 times/minute. This operation is 51! Within I, preferably 2 to 3 reli, repeat 47. In the column method, a highly concentrated lower alcohol aqueous solution of 1 to 1000 times the volume of the gel beads is applied to the ultraviolet detector.
The flow was continued until the absorbance at 0 nm decreased and became constant, and the release was reduced to {1. 1q of gel beads contains 60-95% Q of lower alcohol used for desorption of catechins.
] Use an aqueous solution and finally wash with water to finish. In the batch method, a lower alcohol aqueous solution containing 1 to 5 ∆mold of the volume of gel beads is used, and the washing time is 24 minutes or more.
Within 1 hour, by stirring at 10 to 1000 revolutions/min or milling at 3 to 500 revolutions/min. Repeat this operation 2 to 3 times. Finally, the gel beads are washed with water and regenerated. In the column method, after the lower alcohol aqueous solution is filtered out and the absorbance at 280 nm on the ultraviolet detector decreases and becomes constant, the lower alcohol aqueous solution is continued to flow at 10-1000 times the size of the gel bead body. and finally washed with water to obtain +II-grown gel beads. (Example) Next, the present invention will be explained with reference to an example. Example 1 We tracked the time course of the extraction of catechin heads in tea cakes at room temperature by various j1m solvents and their aqueous solutions (81), 8
The catechin extraction rate was 10 in the 0% ethanol aqueous solution and 50% methanol aqueous solution for 3 hours, and in the 80% methanol aqueous solution 1α and 20% acetone aqueous solution for 3 hours.
It became 0%, but 10% and 20% methanol water? 1
With one solution, the catechin extraction rates were 41% and 81%, respectively, even after 24-inch extraction. In addition, caffeine was also extracted into the extract with a high yield in the alcohol aqueous solution. On the other hand, with ethyl acetate and ethyl ether, the extraction rates of catechin I were 5% and 4%, respectively, and almost no catechin I was extracted. Table 1 Extraction with organic solvents and their aqueous solutions Conditions Extraction time (h) Cuff x 4 yield (Catechin extraction I!) 10 Methanol aqueous solution 24 (41%) 81% 20
X meta/-/l, // H (8t) 79
50 methanol/l 3 (100) 10080
%methanol // l(//) 1008
0% ethanol // 3 (//) 79
20% ace 1. //
1(//) 8000%lTV Rf
i L f Le 2+ (5) 100g of tea leaves (
2+n m or more) in ill solvent is 1↑(300
Rotation/min) Extraction Catechin extraction rate: Extraction wheel Example 2 for catechins contained in tea leaves Nonionic adsorption resin, Amberlite XAD-2, XA
D-4, XAD-7, XAD-8,
II), Duolite S-587, S-7431, S
-862 (Duolite is an adsorption tree made by Dow Chemical.
), Organic solvent resistant gel r Toyokawa filler, Toyovar H
W-40 and Sephadex LI1-20 were each swollen in water with a body diameter of 100 m I.
25 g of tea ethanol 1111 Il + lyophilized product was dissolved in 200 ml of deionized water and adsorbed for 1 hour while stirring at 200 rpm. 31! f&, Washed with deionized water until the color of water disappeared (approximately 600 ml).
Next, 80% ethanol water 72lf liquid 2 0 0 + n
M was added, and desorption was performed for 1 hour with IM stirring at 3001 + 4 J rotation/min.
Washed with 80% ethanol. The last operation is 3
I repeated the circle. In other words, it was {1 in batch mode. Departure II
After removing the ethanol in the liquid using an evaporator, it was freeze-dried and weighed. The moisture content was approximately 5%● Table 2 shows! The m (total adsorption ■) of the kimono, the total catechin m contained therein, the purity of catechin, and the caffeine content are shown. Amparato XAD-8 and XAD- in full absorption iffl
10 is common, but all catechin dishes are Sephadec 7. L
II-20, Elbar II W-4
It was 1'1 degree, the same as 0, and impurities were also adsorbed to Taka. It 1: On the other hand, Sephadex L II-2
0 and Toyobar II W-40 had high catechin purity and less adsorption of impurities such as caffeine. In particular, Toyobar II W-40 specifically absorbed catechin. Table 2ryA arrival 4#4 open etc.'? 3.45 g of catechin nods eluted from ft leaves.
l. ll$g// XAD-4 7.5
5 4.4+1//XAD-7
1.25 4.7? //XAD-
It l. 08 L 87//I
AD-18 10.04 6.11f'Suolite 15117 4.0 1+11//
s-tg+ 4. go LN/
/ S-11N? ．． 44 4.
71 to 3 h-k 11140? ,eye
6.8ISe7rf゜TsukuX Llli0 11
．． 54? ．． HSL? X 111. ! X Ill 1.7 74.1 7.2 75.2 10.5 61. I N. 8 73.8 1.2 57.1 +6. I N. 0 14.4 14. I L2 io. Chewing 5.2 Catechin purity ← Total catechin 11t/Total amount of catechin, including h7xin m = Power 71inIn/Total intake t?
Eyes, full power N冫m is iron tartrate method, power 7x {/Jli is ll
By removing the PLcf, the fixed plate 1 H-Light and T-EOL are made of non-ionic n-absorbing resin.
To set 7T tex, Gel V Flow Charger Example 3 Toyobar H W-4 0, 100ml, 1
Approximately 200 mml of deionized water = ethanol extract of O area
The FB was adsorbed while stirring at 200 rpm for 1 hour. Adsorption {Apply suction with paper and wash with deionized water until the color of the water disappears (approximately 600 mg). Next, the specimens were desorbed with 2 U m It of ethanol of each concentration for 1 hour while being pumped in the same manner, suctioned with a mouthpiece, and washed with 100 m m of ethanol of each concentration. Perform this operation 2. Repeated times. Next, add 80% ethanol 200 mIt to 1:1 and stir 1↑ in the same way.
1 0 0
Washed with 80% ethanol. This operation was repeated three times. The ethanol in the desorption solution was removed using an evaporator, and the sample was then ttm (Table 3).
As a result, when determining the ethanol concentration in which most of the caffeine in the vaping liquid is removed and catechins are less likely to be lost, the following is obtained:
5 was 20%. Furthermore, if the concentration of ethanol to be desorbed is less than 10% (especially more than 5%), caffeine will not be completely desorbed;
At this point, catechins were also removed at the same time as caffeine.
3[! Even after desorption using 50% ethanol, approximately 20% of the catechin mass was still adsorbed to the adsorption resin. 1t3 Concentration of catechins and caffeine due to differences in ethanol concentration Total utt arrival Europe Total catechin chew 5% 4.53g 4.42 animals 10
4.81 4.19I5 4
．． 8g 4.8020 4.3+
4.2425 4.06
3.7430 co. 65
ko, 5G402.302.15 50 1.17 1.09ka 7L
Including Europe 0.27% 0.1l O,01 0.01 0.01 Total cadechin 0.44g o. 33 0,61 0.88 I,83 2,17 Ko,82 4,64 Removal material: Washed with ethanol of various concentrations and removed Adsorbed material: Washed and separated the removed material with 10I concentration ethanol. Example 4 The recovery rate of catechins by washing with a 15% ethanol aqueous solution in Example 3 was as follows:
- 57% (approximately 10 g / 100
mg of gel beads) to 73% (approximately 3 g/100+n
M gel beads), and their catechin purity is approximately l
It was close to 00% and contained almost no caffeine (Table 4
). a4 Recovery rate of catechin and its purity including flm Inhalation t1m Recovery rate Purity Power 7 Amount H
．． 4g 1L41g 56.7% 100%
0.02% 1g. 7 1. G9 g
+1.0 19.4 0.0+1.4
s. 11 N. 4 1L2
Contains 0.01! ! : Total catechin concentration in the adsorbed ethanol extract Example 5 The catechin composition was measured by high performance liquid chromatography (Table 5). As a result, when comparing the catechin composition in the ethanol extract, it was found that the catechin composition in the Toyobar HW-40 adsorbate,
Inhalation with 5% ethanol to remove caffeine, etc.! At t1#, there is less reduction of (-) one bigallocatechin (EGC) and {-}12 bigallocatechin (EGC).
GCG) and (-) ichieki tekin gallate (ECG), on the other hand, increased. 15% washing from ethanol extract
Calculating the recovery rate of the removed adsorbate, (1) one shrimp gallocatechin was 30%, (-) one shrimp gallocatechin gallate was 77%, and one yen one epicatechin gallate was 100%. Table 5 Composition of catechins in extracts and adsorbates (%) Catechins GA gGc KGCG gc
G ethanol extract 1. +26. L
66.8 1.0 to 38”-k adsorption 1 course 0.5 stitches.3 74.8 8
．． 6IS% cleaning/adsorbent 0.5 11.7
78.1 9.6G/M Gallic T monoacid, EGC
: (-) One shrimp gallocatechin, E G C G;
(−) Shrimp gallocatechin gallate, E c
a: (1) Ichieki Riki Tekin Gallate 15% washing/adsorbent: Toyobar H W-4 0
Comparative example 1 of absorbent was washed with 15% ethanol and desorbed with 80% ethanol.
Add 20 g of ethanol extract to approximately 200 m of deionized water.
It was dissolved in M and adsorbed while stirring at 200 rpm for 1 hour. The sample was suctioned with suction paper and paper, and washed with deionized water until the color of the water disappeared (approximately 600 summer n9).
Next, 30% ethanol 200 In Q″c1
The sample was desorbed with stirring for 1 hour, suctioned with a mouthpiece, and washed with ethanol at various concentrations of l O O m H. This operation was repeated twice. Next, in the same manner as I! for 1 hour with 200ml of 80% ethanol. +'?
Detach it from L and suck it with the mouth paper r3At,, l O
Washed with 80% ethanol of O+nQ. This operation was repeated three times. Ethanol in the desorption solution was removed using an evaporator, and then freeze-dried and smoked (Table 6). As a result, Amberlite XAD-16 was unable to selectively remove caffeine using the difference in alcohol concentration. Similar results were obtained for other nonionic absorbent fl. 11 fats, Amberlite XAD-7 and XAD-8. Table 6 Catechin and caffeine in the desorbed product due to changes in ethanol concentration of 7-mparato 11.6%94
47. 4 13. 0 extraction 1 ka =
Extract g1 pickled with 30% ethanol: 30%
Ethanol Extract with 90% Ethanol (Effects of the Invention) According to the present invention, catechins can be efficiently produced on a commercial scale using plants, especially tea, as raw materials. Since these catechins contain almost no impurities such as caffeine, they can be used not only as natural food additives, but also as a supplement for children's snacks and powders. ．．

Claims (3)

[Claims] (1) (a) A step of extracting catechins from a plant using an aqueous solution of lower alcohol, (b) A step of adsorbing the extract to gel beads, (c) Washing the gel beads adsorbed with the extract with water. (d) a step of liberating impurities other than catechins in the gel beads adsorbed matter with a low concentration aqueous solution of lower alcohol; (e) a step of liberating catechins adsorbed on the gel beads with a high concentration aqueous solution of lower alcohol. A method for producing catechins, comprising the following steps. (2) The plant in step (a) is
a￥￥sinensis￥var. siensis, v.
ar. 2. The method according to claim 1, which is a plant of the genus Canellia containing catechins (e.g., Canellia assanica) and a hybrid thereof, and its leaves and other tree constituents, and processed products thereof. (3) The catechins in step (a) are (+)-catechin,
(-)-epicatechin, (-)-epigallocatechin, gallic acid, mono- and digallate esters of (-)-epigallocatechin, mono- and digallate esters of (-)-epicatechin, and further these compounds. The method according to claim 1, which is a double or triple polymer or composite comprising as a constituent component.