JP6838858B2 - Functional agent - Google Patents

Description

The present invention is a functional agent having an antioxidant activity (or a functional agent having other activities in addition to the antioxidant activity) containing an extract separated and obtained from the "leaf part" of a Theaceae plant through a specific step as an active ingredient. It is about.

(Patent Document 1)
The following inventions are shown in claims 1 to 3 of Japanese Patent Application Laid-Open No. 2014-205658 (Patent Document 1) according to the application of the present applicant. Since there are some points that are confusing with the constituent requirements of the present invention, they are quoted in a little more detail.

-1-
Claim 1: A Maillard reaction inhibitor, which comprises a vacuum-dried product obtained by carbonizing a plant raw material under reduced pressure conditions as an active ingredient.
As a reminder, the above-mentioned "dry distillation product" is "distillate liquid distilled from the top of the vacuum carbonization device".

-2-
The claim 2: The vacuum carbonization product is obtained by carbonizing a plant material in a dry or non-dry state under a temperature condition in the range of room temperature to 350 ° C. and a pressure reduction condition of 100 mmHg or less. The Maillard reaction inhibitor according to claim 1, wherein the product is a product.

-3-
3. The Maillard reaction inhibitor according to claim 1 or 2, which is a pentosidine production inhibitor.

-4-
In that paragraph 0025, as examples of plants used as raw materials, "ginger, tea, green onion, moon peach, kakinoha, igusa, shisoyo (shiso leaf), keihi, oregano, chamomile, sage, thyme, basil, peppermint, Lavender, lemon balm, rosemary, shikuwasa, rose, noni, fir tree, rice bran, buckwheat, wheat bran, rapeseed oil cake, camellia oil cake, sesame oil cake, sesame, sardines, keihi, kumazasa, bamboo, green onion, sudachi, autumn corn, Yuzu, Casablanca, Mikan, Jabara, etc. "

-5-
Paragraph 0038 describes the preparation of vacuum dry distillates from raw material plants. That is, the plant is placed in a container, purified water is added, and then heated at 220 ° C. for 4 hours under normal pressure to obtain a “normal pressure distilled liquid”, and then the liquid in the container is further lowered to 220 ° C. under normal pressure. The water-rich liquid distilled at this time is discarded, and then the liquid in the container whose water has been reduced as described above is heated at 300 ° C. for 4 hours under a reduced pressure condition of 25 to 30 mmHg. A dry distillation operation is performed, and a "vacuum dry distillation solution", which is the liquid distilled by this operation, is obtained.

-6-
After that paragraph 0039, the yield of "normal pressure and vacuum distillation solution" when various plants described above were used as raw materials, and the inhibition rate of those "normal pressure and vacuum distillation solutions" (pentosidine production inhibition rate). ), Cleavage rate (AGE-protein cross-linking product model cleavage activity) and the like are shown.

-7-
Examples of the examples: Table 1 shows the yields of the atmospheric distillation solution and the vacuum distillation solution of tea, Table 2 shows the inhibition rate of the atmospheric distillation solution and the vacuum distillation solution of tea (pentocidin production inhibition rate), and Table 3 shows. Is the inhibition rate of the vacuum distillation solution of tea (pentocidin production inhibition rate), Table 4 shows the yield of the vacuum distillation solution of tea, Table 5 shows the inhibition rate of the vacuum distillation solution of tea, and Table 6 shows Tables 2 and 5. A comparison of the inhibition rates of tea in Japan is shown. Table 8 shows the inhibition rate of the vacuum dry distillate of tea as a reference example.

-8-
By the way, the "normal pressure distilled solution" in Patent Document 1 (see paragraph 0038 of the distillate) refers to 200 g of a cut raw material plant in a container, 400 g of purified water, and then at "220 ° C." under normal pressure. This is a method of obtaining a distillate by performing atmospheric distillation by heating for 4 hours. Water was added for steam distillation.
The term "decompressed dry distillation solution" (see paragraph 0038 of the same) refers to the above-mentioned atmospheric distillation in which the solution remaining in the container is heated under atmospheric pressure to dispose of the distillate water-rich solution, and the water content is reduced as such. This is the “liquid to be distilled off” when the liquid in the container is subjected to the vacuum carbonization operation of heating at 300 ° C. for 4 hours under a reduced pressure condition of 25 to 30 mmHg.

-9-
As described above, in Patent Document 1, no interest is paid to the residue remaining in the carbonization apparatus during carbonization under reduced pressure (that is, "can residue"), and there is no explanation. Such canned residue is treated as waste, as in the case of ordinary distillation.

(Patent Document 2)
-1-
Claim 1 of Japanese Patent Application Laid-Open No. 2011-006623 (Patent No. 5437709) (Patent Document 2) according to the application of the present applicant was obtained by carbonizing mugwort under reduced pressure conditions of 260 to 350 ° C. and 100 mmHg or less. Antioxidants containing a dry distillation solution are shown.
As a reminder, the vacuum carbonization in Patent Document 2 is related to "vacuum carbonization at a high temperature of 260 to 350 ° C." Further, the "dry distillation solution" in this document 1 is a "distillate distillate from the top of the column" at the time of "high temperature vacuum carbonization".

-2-
In Tables 1 and 2 of Patent Document 2, as Example 1, the antioxidant capacity of the dry distillation solution obtained by carbonizing "mugwort" at a high temperature of "300 ° C., 25 mmHg" is equivalent to "1003 nmol α-tocopherol". It has been shown to be "quantity / assay".
In addition, in Table 2 of the paragraph 0021 and its paragraph 0022, "When the mugwort was carbonized at 220 ° C., the antioxidant capacity was 15 nmol α-tocopherol equivalent / assay, and when the same mugwort was used as the raw material. However, it was found that the antioxidant capacity was significantly different depending on the difference in carbonization temperature. "

-3-
In Table 1 of Patent Document 2, as Comparative Example 1, the antioxidant capacity when "tea" is used under the same conditions as the above-mentioned mugwort (300 ° C., reduced pressure condition of 100 mmHg or less) is "301 nmol α". -Tocopherol equivalent / assay "has been shown.
The antioxidant capacity of the distillate when tea is carbonized at high temperature under reduced pressure is only 30% of that of mugwort.
Further, according to Table 1 of Patent Document 2, it can be seen that even in the various plants of Comparative Examples 2 to 15, the antioxidant ability is considerably lower than that of mugwort.

-4-
As described above, the invention described in Patent Document 2 relates to "distillate distillate from the top of the column" during carbonization under reduced pressure at "high temperature".
Further, in Patent Document 2, as in Patent Document 1 described above, no attention is paid to the "residue (that is," can residue ") remaining in the carbonization apparatus during carbonization under reduced pressure.

JP-A-2014-205658 JP 2011-006623

-1- (About Patent Document 1)
In Patent Document 1, as stated in claim 1, "a Maillard reaction inhibitor characterized by containing a vacuum-dried product obtained by carbonizing a plant material under reduced pressure conditions as an active ingredient." Although there is a description about "distillate liquid distilled from the top of the carbonization device", no attention is paid to the residue (can residue) remaining at the bottom of the column during carbonization under reduced pressure.

-2- (About Patent Document 2)
The invention described in Patent Document 2 is limited to the "distillate distilled from the top of the column" during carbonization under reduced pressure at "high temperature".
Similar to Patent Document 1, in Patent Document 2, no attention is paid to the residue (can residue) remaining at the bottom of the column during carbonization under reduced pressure.

-3- (Basic differences from Patent Documents 1 and 2)
The inventions described in Patent Documents 1 and 2 are fundamentally different from the inventions of the present application in terms of points of view and technical ideas, without needing to compare and contrast the superiority or inferiority of the action and effect.

The functional agent having an antioxidant activity of the present invention is
The component that is distilled when a water-containing plant material is carbonized under low-temperature and low-pressure conditions using a carbonization device equipped with a decompression mechanism is defined as "low-temperature carbonization distillation (D)" and its low-temperature low-pressure dry distillation operation. When the dry powder or flake-like can residue that remains in the carbonization device later is referred to as "low temperature vacuum carbonization residue (R)",
The plant material is the leaves of Theaceae plants, and
The active ingredient is the extract (E) obtained by extracting the low-temperature carbonization residue (R) of the leaves of the Theaceae plant with a solvent.
It is characterized by.

(About the specificity of the idea and the effect)
-1-
As in Patent Document 1, it is a technical convention that the component of interest (referred to as "f") in the raw material plant is contained in the fraction (distillate) distilled from the top of the column by distillation. The solid residue (can residue) remaining in the carbonization device should be treated as waste.

-2-
Since a part of the component f of interest that should be transferred to the distillate by the distillation operation may remain on the residue (can residue) side, it is not possible to recover the component f of interest from the residue side as much as possible. Although it is conceivable, the component f of interest recovered from the residue is inferior in purity and may contain impurities and colored substances. Therefore, in cases where it is expected to be used on the human body, such recovery is usually not performed. is there.
Even when it is recovered, it is necessary to carry out a refining process on the recovered product, which is rather expensive in terms of cost.

-3-
The applicant has been commercializing the distillate obtained by carbonizing tea leaves under reduced pressure at high temperature (sold under the registered trademark of "Fresh Shiraimatsu" for a wide range of applications requiring deodorant properties). Since the residue (can residue) after carbonization at high temperature and reduced pressure is a carbonized raw material leaf as it is, there is no choice but to dispose of it. Rather, it was difficult to dispose of it.

-4-
By the way, when the tea leaves were "carbonized under low temperature and reduced pressure conditions", the aim was to use the distillate from the top of the tower as a product. The residue (can residue) from carbonization may be available. " It is the intuition of those involved in research.
Therefore, "try rather than think" and "even if the success rate is zero, it is meaningful to be able to confirm that (there is a harvest)", and the "residue (can residue) itself" and " When the "extraction of the residue with a solvent" was examined, and in the latter case, the solvent was also examined in various ways. Rather, it turned out that excellent results can be obtained (rather, much better results can be obtained).

-5-
As in the examples described later, with respect to the "antioxidant activity", the activity of the distillate of the low-temperature carbonization distillation is practically zero or close to zero, whereas the above-mentioned "low-temperature carbonization residue solvent" is used. The "extracted portion" is considerably high, and in particular, the extracted portion of the above residue (can residue) when 50% by volume of ethanol water is used as the solvent shows extremely high "antioxidant activity" as in the examples described later. It was.
In addition, in addition to antioxidant activity, Maillard reaction inhibitory activity (inhibition of pentosidine production), AGE-protein cross-linking product model cleavage activity, and tyrosinase inhibitory activity are also the residue (can residue) after carbonization at low temperature and reduced pressure. It was found that the solvent-based extract often gives favorable results.

(Plant raw material)
-1-
In the present invention, the leaves of Theaceae plants are used as plant raw materials. This is because when the leaves of Theaceae plants are used, a target product containing an active ingredient according to the object of the present invention can be obtained. However, even if some parts other than the leaves (stems, etc.) are mixed, there is no particular problem.
-2-
Examples of Theaceae plants include tea, camellia, ternstroemia, southern mosquito, cleyera japonica, eurya japonica, camellia japonica, stewartia japonica, and tall stewartia.
Among these, tea is particularly important in terms of action and effect (antioxidant activity), and any of the first tea, the second tea, the third tea, and the fourth tea is preferably used. Since even the fourth tea can be preferably used, it is advantageous in terms of the amount of raw materials obtained, the period of raw materials obtained, and the cost of obtaining raw materials.
As for tea types, there are ordinary sencha, gyokuro tea / tencha, tamaryokucha, black tea, etc., and the number of varieties belonging to each tea type is also appropriate. And varieties are used. For example, one example of the variety is "Yabukita", which is mainly used for ordinary sencha, and another example of the variety is "Benifuuki", which is mainly used for black tea and semi-fermented tea.
Powdery parts produced as a by-product in the tea making process, products not commercialized or recovered in the distribution process of tea products, etc. can also be used.
Camellia is also useful, though not as much as tea, because it gives favorable results (see Examples below).

(Conditions for moisture content during carbonization at low temperature and reduced pressure)
In the present invention, the leaves of a water-containing Theaceae plant are carbonized under low temperature and reduced pressure conditions by using a carbonization device provided with a decompression mechanism. However, if the water content is too high, it takes a long time for carbonization and lacks industriality. Therefore, it is desirable that the water content is about 90% by weight or less, particularly about 70 to 80% by weight. Incidentally, the water content of raw tea leaves is often, for example, about 70 to 80% by weight, and the water content gradually decreases when left unattended.

(Temperature and pressure conditions during carbonization at low temperature and reduced pressure)
-1-
As the temperature condition for carbonization at low temperature and reduced pressure, a low temperature in the range of 60 to 20 ° C. is suitable, a more preferable range is 55 to 25 ° C., and a more preferable range is 50 to 30 ° C.
When carbonization under reduced pressure is performed under conditions where the temperature condition exceeds 60 ° C., even if the powder or flake-like residue remaining in the carbonization apparatus is extracted with a solvent, the amount of the active ingredient in the extracted portion (E) is small. In addition, the antioxidative function and other functions (such as the Maillard reaction suppressing function) when the obtained extract (E) is used tend to be insufficient.
-2-
The pressure condition (decompression condition) for carbonization at low temperature is preferably -88 kPa or less (-660 mmHg or less), usually -96 to -100 kPa (-720 to -750 mmHg) in gauge pressure notation.
In absolute pressure notation, it is preferably 13.3 kPa or less (100 mmHg or less), usually 1.3 to 5.3 kPa (10 to 40 mmHg).
If the degree of decompression becomes looser than the above range (if the degree of decompression is insufficient), it will take a long time for carbonization, while if the degree of decompression is too large, there are restrictions on the vacuum system, so both are industrial. It will lack sex.
-3-
By carbonization at low temperature and reduced pressure under the above-mentioned conditions, the desired product can be obtained industrially and efficiently.

(Low temperature vacuum carbonization residue (R), extract from the residue (E))
-1-
When the low-temperature carbonization is performed as described above, the dry particulate can residue remaining in the dry distillation apparatus after the low-temperature carbonization operation is the "low-temperature vacuum carbonization residue (R)", and the solvent from the residue. The "extracted component (E)" according to the above is the object of the present invention.

-2-
Here, various solvents can be used as the solvent, but water, ethanol, or a mixture of water and ethanol is preferable in consideration of obtaining a functional agent applicable to the human body. In particular, a mixed solvent of water and ethanol having a weight ratio of ethanol of 20 to 80% by volume (especially 30 to 70% by volume) (that is, "ethanol water") is optimal.

-3-
The former "low temperature vacuum carbonization residue (R)" itself becomes a product, but the purchaser of the product uses the residue (R) to extract with a solvent to obtain an extract (E), and the extraction thereof. It is also possible to manufacture and sell secondary products and tertiary products by using the fraction (E) by itself, and to further sell the extracted portion (E) to a third party.

-4-
The latter "extracted from the low-temperature carbonization residue (R) with a solvent (E)" not only has excellent antioxidant activity, but also has excellent Maillard reaction inhibitory activity (pentosidine production inhibitory activity) and AGE-protein cross-linking formation model. Since it exhibits preferable activities in terms of cleavage activity and tyrosinase inhibitory activity, it is extremely useful as a functional agent having these activities (functions) at the same time.

(Low temperature decompression dry distillation liquid (D))
Although the above-mentioned carbonization liquid during low-temperature carbonization (low-temperature carbonization distillation (D)) is not the object of the present invention, it may be used for other purposes because it contains an antibacterial component and an aroma component. It can be done and has the advantage of not being wasted.

Next, the present invention will be further described with reference to Examples and Reference Examples.

(Preparation of plant materials)
-1-
As a plant raw material, raw tea leaves (raw tea leaf A) of the fourth tea "Yabukita", which is mainly used for ordinary sencha, were prepared. The reason why we used No. 4 tea as raw tea leaves is that only No. 4 tea was available at the start of this test (September). (If good results are obtained with No. 4 tea, at least the same or better results can be obtained when using No. 1 tea, No. 2 tea, and No. 3 tea, in comparison with the case of using No. 4 tea. It can be expected that it will be.)
In subsequent experiments, raw tea leaves (raw tea leaves B) of "Benifuuki", which are mainly used for black tea and semi-fermented tea, were used as plant raw materials.
These prepared raw tea leaves A and B were literally in the shape of whole leaves, and their water content was 74.7% and 75.0%, respectively. The raw tea leaves were subjected to carbonization at low temperature and reduced pressure, which will be described later. (By the way, when this raw tea leaf is dried, it becomes fine crushed pieces, and when the crushed pieces are crushed with a coffee mill, it becomes fine powder.)
-2-
In addition, as plant raw materials for comparison, blue mandarin orange (squeezed fruit slag, water content is about 80% by weight), blue jiso (leaf part), bitter melon (leaf part), Iyokan (squeezed fruit slag), lemon ( Fruit squeezed slag) and bitter gourd (fruit) were prepared. The water content of these plant raw materials was in the range of 60 to 95% by weight.

(Low temperature vacuum carbonization operation)
In the tank of the tank-shaped carbonization device equipped with a decompression mechanism, the water-containing plant raw material (raw plant raw material) prepared above is put into the tank, and the pressure is reduced (at a gauge pressure) at a low temperature (35-40 ° C) under stirring. The carbonization operation was carried out under the conditions of −98 kPa (-735 mmHg) and 3.3 kPa (25 mmHg) at absolute pressure. The carbonization time was approximately 8 hours, excluding the time required for charging the raw materials, the time required for removing the residue in the tank, the time required for depressurization, and the time required for returning to normal pressure.
These low-temperature vacuum carbonization operations were performed using the same vacuum carbonization device (experimental device with an overall height of about 1.5 meters).

("Residue (can residue) (R)" and "Extract of the residue (can residue) (R) with various solvents (E)" obtained by carbonization at low temperature under reduced pressure)
-1-
What are the properties of the "residue (can residue) (R)" when each of the above eight types of "raw tea leaves A, B, blue oranges, blue jiso, bitter melon, Iyokan, lemon, bitter gourd" is carbonized at low temperature and reduced pressure? Was also in the form of dry powder (powder or flakes).
-2-
For the obtained 8 types of "residues (can residue) (R) in the form of dry powder (powder or flakes)", weigh 5.0 g of each, add 50 mL of the following extraction solvent, and then bring to room temperature. Extraction was performed by stirring for 4 hours.
-Extraction solvent 1: Ethanol-Extraction solvent 2: 50% by volume ethanol water (hereinafter abbreviated as "50% ethanol water")
-Extraction solvent 3: Water With this, each of the above eight types of "residues (can residue) (R) in the form of dry powder (powder or flakes)" is extracted with each of the extraction solvents 1, 2 and 3. 24 kinds of "extracted components (E)" (8 kinds x 3 kinds = 24 kinds) were obtained.

-3-
("Distillate (D)" obtained by carbonization at low temperature and reduced pressure)
In addition, when each of the above eight kinds of plant raw materials was carbonized at low temperature under reduced pressure, "eight kinds of distillates (D) distilled from the top of the tower" were also recovered, and various antioxidant activities were collected. It was used for comparison with the above-mentioned -2- "extracted component (E)" regarding activity.

(Measurement item)
The measurement items are as follows.
(Part 1) Antioxidant activity (Part 2) Maillard reaction inhibitory activity (inhibition of pentosidine production)
(Part 3) AGE-protein cross-linking product model cleavage activity (Part 4) Tyrosinase inhibitory activity (whitening)

(Part 1) Antioxidant activity (1) Method (1-1) Preparation of 400 μM DPPH (1,1-diphenyl-2-picrylhydrazyl) ethanol solution Weigh 3.94 mg of DPPH and dissolve in 25 mL of ethanol by stirring.

(1-2) Preparation of calibration curve a: A 400 μM DPPH ethanol solution is diluted 3-fold with ethanol, and 0.9 mL of the diluted solution is dispensed into a test tube.
B: Add 300, 250, 200, 150, 100 μL of ethanol to the test tube containing the diluent at n = 2.
C: Add 0.2 mM α-tocopherol ethanol solution (8.6 mg → 100 mL of ethanol) to the test tube (n = 2) so that the total of ethanol added in B and 0.2 mM α-tocopherol ethanol solution becomes 300 μL. , Stir. At this time, the amount of α-tocopherol is 0, 10, 20, 30, 40 nmol / assay.
D: 20 minutes after addition, the absorbance is measured at 516 nm.
E: Plot the amount of α-tocopherol (nmol / assay) on the horizontal axis and the absorbance on the vertical axis, and prepare a calibration curve by the method of least squares.

(1-3) Measurement of test solution a: 0.9 mL of DPPH diluted solution is dispensed into a test tube, and 250 μL of ethanol is added thereto.
B: Add 50 μL of the test solution to the test tube (n = 2) and stir.
C: 20 minutes after addition, the absorbance is measured at 516 nm.
D: From the obtained absorbance, calculate the equivalent amount of α-tocopherol using a calibration curve.

(Part 2) Maillard reaction inhibitory activity (inhibition of pentosidine production)
-1-
The reaction solution having the composition shown in the table below was prepared in a plastic tube having a capacity of 1.5 mL and incubated on a heat block at 60 ° C. for 24 hours.

(Table 1)

Reaction solution composition Addition amount
50 mM ribose 100 μL
50 mM lysine 100 μL
50 mM arginine 100 μL
100 mM Na2HPO4 (pH 7.4) 100 μL
Sample solution or Blank 100 μL

-2-
After completion of the reaction, 400 μL of purified water was added to 100 μL of the reaction solution, and the peak area of pentosidine obtained by HPLC analysis of the diluted solution was measured to determine the inhibition rate.
In addition, a 10 mM solution of aminoguanidine hydrochloride was used as a positive control.

-HPLC analysis conditions Column: YMC-Pack ODS A-312
150 x 6 mm I. D.
Eluate: 3% CH3CN / 0.1% TFA
Flow rate: 1.0 mL / min
Column temperature 40 ° C
Detector: Spectral fluorescence detector EX335nm, EM380nm
Injection volume: 20 μL
Holding time: Approximately 12 minutes

-Inhibition rate The inhibition rate was calculated by the following formula.
Inhibition rate (%) = 100-[(peak area of pentosidine in sample solution / peak area of blank pentosidine) x 100]

(Part 3) AGE-Protein cross-linking product model cleavage activity-1-
S. The cleavage activity of the AGE-protein crosslinked product model was measured according to the method of Vasan et al. (Nature, Vol. 382, p275-278, 1966).
That is, the reaction solution having the composition shown in Table 2 below was prepared in a plastic tube having a capacity of 1.5 mL, and shaken at 37 ° C. for 4 hours.
-2-
After completion of the reaction, 200 μm of 2N HCl was added and stirred to stop the reaction.
The solution was filtered through a 0.2 μm filter to obtain an HPLC analysis sample solution.

(Table 2)

Reaction solution composition Addition amount
500 mM Na2HPO4 (pH 7.4) 800 μL
100 mM 1-Phenyl-1,2-Propanedione 100 μL
Sample solution 100 μL

-1-
-HPLC analysis conditions Column: YMC-Pack ODS A-312
150 x 6 mm I. D.
Eluate: 40% MeOH / 0.1% TFA
Flow rate: 1.0 mL / min
Column temperature: 40 ° C
Detector: UV wavelength 273 nm
Injection volume: 20 μL
Retention time: Approximately 12 minutes -2-
-How to determine the cleavage rate The cleavage rate was calculated according to the following formula because it can be assumed that 10 mM benzoic acid is liberated when all 1-phenyl-1,2-propanedione is cleaved.
Cleavage rate (%) = (peak area of benzoic acid generated from each analytical sample / peak area of 10 mM benzoic acid) × 100

(Part 4) Tyrosinase inhibitory activity-1-
A. 30 mM phosphate buffer (pH 6.8) 1.8 mL,
1.66 mM L-tyrosine solution 1.0 mL,
0.1 mL of each sample solution
Was mixed and preheated for 5 minutes in a constant temperature bath at 37 ° C.
-2-
I. After adding 0.1 mL of the tyrosinase solution, the mixture was stirred and heated again in a constant temperature bath at 37 ° C. for 10 minutes.
-3-
C. 0.1 mL of 1M sodium azide was added, and the absorbance at 475 nm was measured to calculate the tyrosinase activity inhibition rate (%). A 3 mM arbutin solution was used as a positive control.

(About each measurement item and result)

The results for the above measurement items are shown in Tables 3 to 5 below.
Table 3: Antioxidant activity Table 4: Maillard reaction inhibitory activity (inhibition of pentosidine production)
Table 5: AGE-protein cross-linking product model cleavage activity Table 6: Tyrosinase inhibitory activity

(Table 3)

(1) Antioxidant activity

Antioxidant activity (n mole α-tocopherol equivalent / assay)
Plant name Extraction of dry distillation residue (can residue) with solvent (E) Distillate (D)
Ethanol 50% ethanol water water
Blue mandarin orange 20 133 121 0
Blue Jiso 37 41 60
Camellia 1028 1856 42 0
Iyokan 8 106 9400
Lemon 34 230 1940
Bitter gourd 6 53 77 2
Raw tea leaves A 1210 3990 1290 0
Raw tea leaves B 3180 5930 2190 1

(Note) Extract E: Extraction of low-temperature vacuum carbonization residue (can residue) with solvent (Note) Distillate D: Distillate from the top of the tower during low-temperature carbonization (Note) Raw tea leaf A is " The leaf part of "Yabukita" and the raw tea leaf B are the leaf part of "Benifuuki".

-1-
In addition to the above Table 3, the antioxidant activity of the extract (E) of the dry distillation residue (can residue) of raw tea leaf B with 50% ethanol water is "5930 n mole α-tocopherol equivalent / assay". Is.
This value is the antioxidant capacity of yomogi obtained by drying under high temperature and reduced pressure conditions (1003 n mole α-tocopherol equivalent amount /) in Patent Document 2 described in paragraphs 0012 to 0015 of the specification of the present application. Compared with "assay) and the antioxidant capacity of tea (301 n mole α-tocopherol equivalent / assay)", it is an extremely strong antioxidant activity that can be said to be amazing.
The applicant of Patent Document 2 is the same as that of the present application, and some of the inventors are also common. However, according to the present application, the champion data of the application of Patent Document 2 is rewritten at once.
-2-
In addition, the raw tea leaf B dry powder used in Table 3 above is a can residue (powder in a dry state or flake-like residue) during carbonization at low temperature and reduced pressure, and is on the top side of the column during carbonization at low temperature. It is not a distillate that dives from. Despite the use of such a can residue, as shown in Table 3 above, the antioxidant activity of the 50% ethanol water extract of the can residue is "5930 n mole α-tocopherol equivalent / assay". There was.

(Table 4)

(2) Maillard reaction inhibitory activity (inhibition of pentosidine production)

Blocking rate (%)
Plant name Extract distillate of dry powder (remaining can) (D)
Ethanol 50% ethanol water water
Blue oranges 24 64 92 91
Blue Jiso 12 89 89 85
Camellia 97 99 93 67
Iyokan 7 83 93 86
Lemon 32 49 83 92
Bitter gourd 0 0 51 93
Raw tea leaves A 97 100 99 93
Raw tea leaves B 99 100 99 60

(Note) The inhibition rate of 10 mM aminoguanidine hydrochloride in the positive control was 41% in the assay of blue mandarin orange, blue jiso, bitter melon, Iyokan, and raw tea leaf A, 52% in the assay of bitter gourd, and raw tea leaf B. It was 45% at the time of the assay.
(Note) Raw tea leaf A is the leaf part of "Yabukita", and raw tea leaf B is the leaf part of "Benifuuki".

(Table 5)

(3) AGE-Protein cross-linking product model cleavage activity

Cutoff rate (%)
Plant name Extract distillate of dry powder (remaining can) (D)
Ethanol 50% ethanol water water
Blue oranges 3 4 4 7
Blue Jiso 3 4 4 2
Camellia 7 8 6 2
Iyokan 4 5 5 2
Lemon 7 11 10 6
Bitter gourd 8 9 9 8
Raw tea leaves A 10 32 15 2
Raw tea leaves B 16 24 12 2

(Note) The cleavage rate of 10 mM-PTB of the positive control was as follows.
When assaying blue oranges, blue jiso, camellia, and Iyokan: 40%
During the lemon assay: 43%
During bitter gourd assay: 46%
When assaying raw tea leaf A: 39%
When assaying raw tea leaf B: 44%
(Note) Raw tea leaf A is the leaf part of "Yabukita", and raw tea leaf B is the leaf part of "Benifuuki".

(Table 6)

(4) Tyrosinase inhibitory activity

Blocking rate (%)
Plant name Extract distillate of dry powder (remaining can) (D)
Ethanol 50% ethanol water water
Blue mandarin orange 3 3 0 2
Blue Jiso 0 1 0 0
Camellia 0 0 0 2
Iyokan 0 0 0 0
Lemon 0 0 0 0
Bitter gourd 0 1 3 0
Raw tea leaves A 0 14 4 0
Raw tea leaves B 3 18 0 0

(Note) The dry powder extract was tested using a 10-fold diluted solution.
(Note) Raw tea leaf A was the leaf part of "Yabukita", and raw tea leaf B was the leaf part of "Benifuuki". (Note) The inhibition rate of 3 mM arbutin in the positive control was as follows.
When assaying blue oranges, blue jiso, camellia, and Iyokan:
・ Ethanol extract is 20%
・ 50% ethanol water extract is 17%
・ 18% of water extract and distillate
Lemon, raw tea leaf A assay:
・ Ethanol extract is 18%
・ 50% ethanol water extract is 19%
・ 20% of water extract and distillate
During bitter gourd assay:
・ Ethanol extract is 23%
・ 50% ethanol water extract is 20%
・ 19% of water extract and distillate
When assaying raw tea leaf B:
-Ethanol extract and 50% ethanol water extract are 23%
・ 17% of water extract and distillate

The functional agent of the present invention containing the solvent-based extract (E) from the low-temperature vacuum-dried residue (R) of the leaves of Theaceae plants as an active ingredient has an outstanding antioxidant function and a Maillard reaction inhibitory activity (pentosidine). It is useful as an additive to foods, cosmetics, quasi-drugs, etc. because it has preferable functions in terms of production inhibitory activity), AGE-protein cross-linking product model cleavage activity, and tyrosinase inhibitory activity.

Claims (6)

The component that is distilled when a water-containing plant material is carbonized under low-temperature and low-pressure conditions using a carbonization device equipped with a decompression mechanism is defined as "low-temperature low-pressure dry distillation liquid (D)" and its low-temperature low-pressure dry distillation operation. When the dry powder or flake-like can residue that remains in the carbonization device later is referred to as "low temperature vacuum carbonization residue (R)",
The plant material is tea or camellia leaves, and
The low-temperature vacuum carbonization residue (R) of the tea leaves is extracted with water, ethanol, or a mixture of water and ethanol (E), or the low-temperature vacuum carbonization residue (R) of the camellia leaves is ethanol or The active ingredient is the extract (E) when extracted with a mixture of water and ethanol.
An antioxidant characterized by. The component that is distilled when a water-containing plant material is carbonized under low-temperature and low-pressure conditions using a carbonization device equipped with a decompression mechanism is defined as "low-temperature low-pressure dry distillation liquid (D)" and its low-temperature low-pressure dry distillation operation. When the dry powder or flake-like can residue that remains in the carbonization device later is referred to as "low temperature vacuum carbonization residue (R)",
The plant material is tea or camellia leaves, and
The active ingredient is the extract (E) obtained by extracting the low-temperature vacuum-dried residue (R) of the tea or camellia leaves with water, ethanol, or a mixture of water and ethanol.
A Maillard reaction inhibitor characterized by. The component that is distilled when a water-containing plant material is carbonized under low-temperature and low-pressure conditions using a carbonization device equipped with a decompression mechanism is defined as "low-temperature low-pressure dry distillation liquid (D)" and its low-temperature low-pressure dry distillation operation. When the dry powder or flake-like can residue that remains in the carbonization device later is referred to as "low temperature vacuum carbonization residue (R)",
The plant material is tea leaves, and
The active ingredient is the extract (E) obtained by extracting the low-temperature carbonization residue (R) of the tea leaves with a mixture of water and ethanol containing 50% by volume of ethanol.
A tyrosinase inhibitor characterized by. The low-temperature carbonization operation is performed on the leaves of tea or camellia having a moisture content of 90% by weight or less under low-temperature conditions of 60 to 20 ° C. and under reduced pressure conditions of −88 kPa or less at a gauge pressure. The antioxidant according to claim 1 , which is a dry distillation operation. The low-temperature carbonization operation is performed on the leaves of tea or camellia having a moisture content of 90% by weight or less under low-temperature conditions of 60 to 20 ° C. and under reduced pressure conditions of −88 kPa or less at a gauge pressure. The Maillard reaction inhibitor according to claim 2, which is a dry distillation operation. The low-temperature carbonization operation is performed on the leaves of tea or camellia having a moisture content of 90% by weight or less under low-temperature conditions of 60 to 20 ° C. and under reduced pressure conditions of −88 kPa or less at a gauge pressure. The tyrosinase inhibitor according to claim 3, wherein the dry distillation operation is performed.