HK1101135B - Lipase activity inhibitor containing high-molecular weight polyphenol fraction, tea extract and process for producing the same - Google Patents

Description

Lipase activity inhibitor containing high molecular weight polyphenol fraction, tea extract and its preparation method

Technical Field

The present invention focuses on that a high molecular weight polyphenol (polymeric catechin) contained in tea such as oolong tea has various effects and is less astringent and less bitter unlike other tannins and non-polymeric catechins, and relates to a method for selectively separating a polymeric catechin such as tea, a composition such as a tea extract obtained by concentrating the polymeric catechin obtained by the method, a lipase activity inhibitor containing a fraction obtained by the method, and a food or beverage having a high palatability and intended for health promotion without impairing the flavor and palatability when the composition is added.

Background

In recent years, with the westernization of the lifestyle of japanese people, intake of high-fat foods has been increasing. According to the report of national nutrition survey in the first 11 years, although the energy intake is reduced year by year, the ratio of the lipid energy ratio is more than 25%, and the ratio of the neutral fat value and the cholesterol value is higher, namely, the ratio of the lipid energy ratio to the lipid energy ratio, is 5-6 for people over 60 years or over 60 years (the Processary labor is the summary clinical nutrition 2001 of the results of national nutrition survey in the first 11 years; 98 (5): 577-.

Obesity is one of the most important diseases in today's society, and its main cause is excessive intake of fat. In addition, excessive intake of fat may cause obesity, and diabetes, hyperlipidemia, hypertension, arteriosclerosis, etc. may be caused by obesity. The only therapeutic drug for obesity recognized in japan is the appetite suppressant, mazindol r (mazindol), but it has been reported that it has side effects such as thirst, constipation, stomach discomfort, nausea, vomiting, and the like [ clinical evaluation 1985; 13(2): 419-459 clinical evaluation 1985; 13(2): 461-515]. Further, Xenocol R (Xenical), which has The effect of inhibiting The absorption of fat in The intestinal tract by inhibiting lipase activity in overseas, is also marketed as an obesity-ameliorating agent, but is still not safe because of The side effects of fatty stools, increased number of stools, soft stools, diarrhea, abdominal pain, etc. (The Lancet 1998; 352: 67-172).

Although it is effective to reduce the intake of calories by diet control for obesity prevention, strict nutritional guidelines must be accepted, and it is difficult to practice the diet in daily life. Therefore, it is a realistic and effective method to safely and healthily inhibit the absorption of dietary fat into the body and to treat obesity and diseases related thereto or to improve health.

In this context, the development of specified health foods which have proven to be both safe and effective for the human body is attracting attention. Heretofore, as a food material for suppressing the increase in the serum neutral fat value after meal, there has been sold a globin hydrolysate for suppressing fat absorption by suppressing pancreatic lipase [ j.nutr.1988; 128: 56-60, journal of Japan society of clinical and food and diet 1999; 52(2): 71-77 health and nutritional food research 2002; 5(3): 131-144], a diglyceride having a different digestion and absorption property from triglyceride [ J.Am.Coll.Nutr.2000; 19(6): 789-; 11(2): 109-117], eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and the like extracted from fish oil.

Meanwhile, as reports showing lipid-improving effects of oolong tea, there are reports that neutral fat values in blood were significantly reduced after drinking commercial oolong tea for 6 weeks at 1330ml per day (journal of japan nutrition and food society 1991; 44(4): 251-259], and also with respect to a test subject of 102 male and female simple obese persons who took oolong tea orally (2g × 4/day) for 6 weeks, 67% of subjects showed a decrease in body weight of 1kg or more and subjects having a high value of neutral fat in blood showed a significant improvement effect after taking oolong tea [ journal of japanese society for clinical nutrition 1998; 20(1): 83-90]. Such a large amount of oolong tea, though effective, is difficult to sustain in daily life. Furthermore, even if oolong tea is provided as a pure concentrated oolong tea, it is not practical because the bitterness and astringency are too strong and the amount of caffeine increases.

The high molecular weight polyphenol (polymer catechin) contained in oolong tea and the like not only has various effects, but also has little astringency and bitterness unlike other tannins, non-polymer catechins and the like. Therefore, it is highly desirable to establish a method for efficiently separating non-polymeric catechins and polymeric catechins from catechins contained in oolong tea and the like.

It has been known that various components of tea can be separated by various resins. For example, treatment with activated carbon can remove tannins, remove caffeine. However, an effective method for selectively separating the polymerized catechins and the non-polymerized catechins by an adsorbent such as activated carbon or an adsorbent resin has not been known.

[ Nonpatent document 1 ] summary clinical nutrition 2001 of national nutrition survey results, which is well established in the past 11 years by the Min's province of fat birth labor; 98(5): 577-588

[ non-patent document 2 ] clinical evaluation 1985; 13(2): 419-459

[ non-patent document 3 ] clinical evaluation 1985; 13(2): 461-515

[ non-patent document 4] The Lancet 1998; 352: 67-172

[ non-patent document 5] J.Nutr.1988; 128: 56-60, 1988

[ non-patent document 6 ] Japan society of clinical and food and drug sciences 1999; 52(2): 71-77

[ non-patent document 7] research on health and nutritious foods 2002; 5(3): 131-144

[ non-patent document 8 ] J.am.Coll.Nutr.2000; 19(6): 789-796

[ non-patent document 9] Clin. Chim. acta.2001; 11(2): 109-117

[ non-patent document 10 ] Japan society of Nutrition and diet 1991; 44(4): 251-259

[ non-patent document 11 ] journal of the Japanese society for clinical nutrition 1998; 20(1): 83-90

Disclosure of Invention

The present invention provides a food and drink product and a lipase activity inhibitor which inhibit the absorption of fat in the diet, inhibit the increase of neutral fat in the blood, and/or prevent obesity.

The present invention provides a process for efficiently separating non-polymeric catechins and polymeric catechins from a mixture of both, such as oolong tea.

The present invention provides a method for selectively removing non-polymeric catechins by bringing an aqueous liquid containing polymeric catechins and non-polymeric catechins into contact with an adsorbent at a temperature of 50 ℃ or more than 50 ℃ to obtain an aqueous liquid having an increased ratio of polymeric catechins to non-polymeric catechins as compared with the original aqueous liquid, and a composition comprising the aqueous liquid obtained by the method or a concentrate or dried product thereof.

The present invention provides a tea extract, particularly an oolong tea extract, and preferably an oolong tea extract in which the ratio of polymerized catechins to non-polymerized catechins is higher than that of a usual oolong tea (extract).

The invention also provides a lipase inhibitor and a beverage food additive containing the composition or the tea extract.

Definition of

The term "polymerized catechin" as used herein means a substance polymerized with a dimer or more, and is an antisense term for a non-polymerized catechin, which is used synonymously with a high-molecular polyphenol contained in tea. Tea catechins are a generic term for catechins contained in tea, and catechins contained in tea are derived from other raw materials or from synthesis, and are also included in the definition of tea catechins in the present specification.

Non-polymeric catechins refer to monomeric tea catechins. The main nonpolymeric catechin contained in oolong tea is 8 kinds of catechin, gallocatechin, Catechin Gallate (CG), gallocatechin gallate (GCG), epicatechin, epigallocatechin, epicatechin gallate (ECG) and epigallocatechin gallate (EGCG).

The term "high-molecular-weight polyphenol fraction" as used herein refers to an oolong tea extract obtained by selectively concentrating polymerized catechins in an aqueous extract of a material such as oolong tea, which is obtained by mixing polymerized catechins and non-polymerized catechins, wherein the ratio of the polymerized catechins to the non-polymerized catechins is higher than that of the oolong tea extract (aqueous extract).

The adsorbed catechins in the present specification mean tea catechins adsorbed by the adsorbent when the method of the present invention (for example, the method of example 2 in which the temperature is 60 ℃) is carried out, and most of them are nonpolymeric catechins. One skilled in the art will appreciate that the adsorbent will also adsorb some of the polymerized catechins. Non-adsorbed catechins refer to tea catechins which are not adsorbed by the adsorbent, and most of them are polymerized catechins. One skilled in the art will also appreciate that non-adsorbed catechins will also contain some non-polymeric catechins.

(1) Lipase activity inhibitor invention

The present inventors have found that a component of a lipase, particularly pancreatic lipase, necessary for inhibiting fat absorption in oolong tea is obtained by using oolong tea as an adsorbent column of activated carbon or the like, and is present in an oolong tea polyphenol fraction containing none or only a trace amount of non-polymeric catechins, and caffeine recovered as a non-adsorbed fraction. Based on the above findings, the present invention is directed to a lipase activity inhibitor comprising a high-molecular polyphenol fraction isolated from oolong tea as an active ingredient for inhibiting fat absorption, inhibiting the increase of neutral fat in blood, and/or preventing obesity, and a food or drink containing the high-molecular polyphenol fraction.

The high molecular weight polyphenol fraction extracted from oolong tea used in the present invention preferably contains polymerized catechin (high molecular weight polyphenol) in an amount of at least 4 times as much as that of non-polymerized catechin. The method for obtaining the high molecular weight polyphenol component is part of the present invention and is described in detail later.

The lipase activity inhibiting component of the present invention contains no or only a trace amount of non-polymeric catechins, and therefore, basically does not feel bitter or astringent, and also contains no caffeine, so that the flavor is not spoiled when added in an arbitrary amount to a food or beverage, and excessive ingestion of caffeine is not caused even when a large amount is ingested. Furthermore, the component for inhibiting lipase activity of the present invention is derived from oolong tea, and therefore, is highly safe. The food and drink of the present invention can be taken daily or daily, and can exert desired effects continuously. Therefore, there is no substantial upper limit or lower limit of the amount of the polymer polyphenol component to be added to the food or beverage. However, in order to obtain the effect of inhibiting lipase activity, it is preferable that the amount of polymerized catechin ingestible in each portion (for example, about 250ml) to be added to the food or beverage is 67m g or more than 67m g. In this case, the amount of the polymerized catechin contained in the food or beverage after the addition can be measured by the high performance liquid chromatography shown in example 1, in which the fractionated polymerized catechin is used as a standard substance and is subjected to gradient elution by a reverse phase column.

Examples of the beverage and food containing oolong tea high molecular polyphenol component as the component for inhibiting lipase activity include liquid tonic, health beverage, nutritional supplement beverage, and sport beverage. When used as food, it can be used as health food, nutritional supplement food, etc.

Lipase activity inhibitor is a liquid component obtained by contacting an aqueous extract of oolong tea with an adsorbent selected from activated carbon and adsorbent resin to selectively remove non-polymeric catechin, and concentrating or drying the liquid component to increase the ratio of polymeric catechin to non-polymeric catechin.

(2) Method for producing high-molecular polyphenol fraction derived from tea

The present inventors have made intensive studies on a method for separating non-polymeric catechins from polymeric catechins, and as a result, have found a method for obtaining a high-molecular polyphenol fraction (a composition or tea extract having an increased ratio of polymeric catechins to non-polymeric catechins) by selectively concentrating only polymeric catechins by using an adsorbent in which the temperature is controlled to 50 ℃ or more than 50 ℃.

Therefore, the method of the present invention is configured to selectively remove non-polymeric catechins by bringing an aqueous liquid containing polymeric catechins and non-polymeric catechins into contact with an adsorbent selected from activated carbon and an adsorbent resin at a liquid temperature of 50 ℃ or more, thereby obtaining an aqueous liquid having an increased ratio of polymeric catechins to non-polymeric catechins as a high molecular weight polyphenol component as compared with the original aqueous liquid.

Temperature of

The temperature is set to 50 ℃ or more than 50 ℃ which is an essential condition for removing only the non-polymeric catechins by selectively adsorbing them on the adsorbent in the present invention, and below this temperature, for example, the polymeric catechins and the non-polymeric catechins cannot be selectively separated at room temperature. The temperature is not particularly limited, and may be any temperature up to the boiling temperature. Temperatures in excess of 100 ℃ under pressure may also be used, as desired.

Raw materials

The aqueous liquid containing polymeric catechin and non-polymeric catechin is not particularly limited, and the method of the present invention is typically used for efficiently separating polymeric catechin and non-polymeric catechin from a plant extract, for example, an extract of tea, particularly oolong tea. The oolong tea is described below as an example, but not limited thereto.

Pretreatment of

A raw material containing polymerized catechin and non-polymerized catechin, such as oolong tea leaf, is cut into pieces as required, and extracted with water as appropriate. The temperature of water during extraction is not particularly limited, but is preferably 50 to 99 ℃ and more preferably 80 to 99 ℃ in order to improve the extraction efficiency by extracting in a short time. Sodium bicarbonate can be added to make the extractive solution slightly alkaline. Sodium bicarbonate may be added at any concentration from a saturated state without addition. For example, 1.0 to 2.0g of sodium bicarbonate can be added to 1L of warm water, or sodium bicarbonate can be added to a pH of 8.0 to 8.5, preferably about 8.2. Instead of sodium bicarbonate, other weakly basic substances with high safety can also be used. After extraction, for removing solid components, standing, centrifuging and/or filtering, and optionally adding Vitamin C (VC).

The concentration of oolong tea extract contacted with the adsorbent is Brix 2.0-6.0, preferably Brix is about 3.7.

Adsorbent and process for producing the same

The adsorbent used for the separation is selected from activated carbon and adsorbent resin. The adsorbent is preferably adapted for use in a chromatographic column, the choice of particle size of the adsorbent to be subjected to the column is also an important factor. The larger the particle size, the better the pressure loss when using a column, but the smaller the particle size, the more preferable the particle size is to ensure the adsorption surface area of the adsorbent for efficient separation. The selection of the optimum particle size in accordance with the adsorbent used can be carried out within the ordinary skill of the person skilled in the art. As shown in examples, the granular activated carbon is particularly preferably 32 to 60 mesh in size.

As the synthetic adsorbent resin for granular activated carbon, for example, a resin using polystyrene as a raw material can be used. In this case, the pore size is an important factor, and an appropriate resin is selected for efficient separation. Among commercially available resins, Sepabeeds SP825 (average pore diameter 57.4) of Mitsubishi chemical can be preferably used) SP850 (average pore diameter: 38.1)) And the like.

Amount of adsorbent used

In order to efficiently carry out the process of the present invention, it is preferable to use the adsorbent in an amount as large as possible. That is, in the case of the column method, the amount of the oolong tea extract separated by the column is more effective as the amount is smaller, and the capacity is preferably 5 times (5CV) to 10 times (10CV) the capacity of the column. The amount of the oolong tea extract is important in the present invention, provided that the adsorbent is used in an amount of at least a certain amount, in addition to the temperature of 50 ℃ or more during the adsorption treatment.

That is, it is considered that neither polymeric catechin nor non-polymeric catechin is sufficiently adsorbed by the adsorbent, but according to the present invention, non-polymeric catechin can be selectively and sufficiently adsorbed on the adsorbent by maintaining the adsorption treatment temperature at 50 ℃ or more than 50 ℃ and using a sufficient amount of the adsorbent.

Time of contact with adsorbent

The time for contacting the oolong tea extract (aqueous extract) with the adsorbent is not particularly limited as long as the nonpolymeric catechin is sufficiently adsorbed by the adsorbent. When adsorption is performed using a column, the adsorption can be performed at a flow rate of SV of 1 to 6, preferably SV of about 3.

Post-treatment

The oolong tea extract (high molecular polyphenol fraction, composition, tea extract) separated from adsorbent can be used as beverage rich in high molecular polyphenol (polymerized catechin) directly, or used as material for beverage food or medicinal products. If necessary, the extract can be used as a concentrate or a dried product by a heating method, a reduced pressure method, a freeze-drying method, or the like.

(3) A high molecular polyphenol fraction according to the production method

The present invention relates to -containing composition in an aqueous, wet or dry form, which is obtained by the above-mentioned method of the present invention and contains a high-molecular polyphenol fraction having an increased ratio of polymerized catechins to non-polymerized catechins. The composition of the invention is a tea extract, preferably an oolong tea extract. In the composition (tea extract), it is preferable that the ratio of the polymerized catechin to the non-polymerized catechin is at least 4 times.

The ratio of the polymerized catechins to the non-polymerized catechins can be confirmed by measurement by the method described later in example 1.

The commercial oolong tea has a weak effect of suppressing the increase of neutral fat after meal, and if the concentration is simply increased, it is not suitable as a beverage due to the increase of bitterness and astringency and the increase of caffeine amount. The present invention can achieve the effect of suppressing the increase of neutral fat and prevent obesity without impairing the flavor of foods and drinks by separating a large amount of a high molecular polyphenol component containing polymerized catechin contained in oolong tea and adding the same to foods and drinks. In order to suppress the absorption of edible fat, it is preferable to ingest the food at the same time, so that the beverage enriched with the effective component obtained from tea is more significant.

Further, according to the present invention, it is possible to easily perform the operation of selectively separating the polymeric catechins and the non-polymeric catechins, which has been difficult so far.

Drawings

FIG. 1 is a schematic view showing an example of the change of the components of an oolong tea extract liquid at 60 ℃.

FIG. 2-1 is a graph showing a comparison between the change amount (. DELTA.TG) of the neutral fat value in blood with time in the group of intake of water (2E) as the oolong tea polyphenol fraction and the fat load of the water, and FIG. 2-2 shows the area under the curve (AUC) in FIG. 2-1.

FIG. 3-1 is a graph showing the change in the amount of change in neutral fat value (. DELTA.TG) in blood with time after the intake of the polyphenol-enriched oolong tea (OT + E) and the fat load of water, and FIG. 3-2 is a graph showing the area under the curve (AUC) in FIG. 3-1.

FIG. 4-1 is a graph showing the change in the amount of change in neutral fat value (. DELTA.TG) in blood with time after taking up the fat load of water in each of the intake groups of Oolong Tea (OT) and polyphenol-fortified oolong tea (OT +0.5E), and FIG. 4-2 is a graph showing the area under the curve (AUC) of FIG. 4-1.

FIG. 5 shows the fat excretion-promoting effect of oolong tea reinforced by polyphenol.

Examples

Example 1

Extraction of tea leaves

(1) The extraction efficiency of solid oolong tea was compared between the case where 1.67g of sodium hydrogencarbonate (pH 8.22) was dissolved in 1L of warm water at 90 ℃ and the case where no sodium hydrogencarbonate was added (pH 6.79). 100g of oolong tea leaves were added to 1L of warm water, and the mixture was slowly stirred at 90 ℃ for 20 minutes to extract the tea leaves.

When the Brix concentration and pH of the extract were measured, the Brix was 3.76 and the pH was 5.06 in the absence of sodium hydrogencarbonate, and the Brix was 4.27 and the pH was 6.15 in the presence of sodium hydrogencarbonate. Since sodium bicarbonate was used, it was apparent that a concentrated extract was obtained, and the improvement in the extraction of the solid matter was confirmed from the Brix value.

(2) 1.67g of sodium bicarbonate was dissolved in 1L of warm water at 90 deg.C, and 100g of oolong tea leaves was added. Extraction was carried out while keeping at 90 ℃ and stirring slowly for 5 minutes. After extraction, the tea leaves and the extract were separated by a 140-mesh sieve, and the extract was centrifuged to remove fine solid components. The separated liquid was added with 1.59g of VC to obtain an oolong tea extract having Brix of 3.7.

(3) Method for measuring polymerized catechin and non-polymerized catechin using Develosil C30 UG-3 (C)Nomuram chemical co.), mobile phase: a: 0.1% HCOOH/H2O, B: 90% CH3CN, 0.1% HCOOH/H2O, 0.2ml/min, gradient: b10% → 70% (15min), B70% iso (10min), detection was performed A280 nm.

The standard substance was polymerized catechin fraction repeatedly fractionated by ODS column chromatography for fractionation from oolong tea extract.

The components of the extract are shown in Table 1.

TABLE 1

^*A non-polymeric catechin; total of catechin, gallocatechin, Catechin Gallate (CG), gallocatechin gallate (GCG), epicatechin, epigallocatechin, epicatechin gallate (ECG), and epigallocatechin gallate (EGCG)

Example 2

Separation of

50ml of a commercially available granular activated carbon (for example, GW-H32/60 manufactured by Kuraray Co., Ltd.) was degassed and packed in a column. The oolong tea extract obtained in example 1 was passed through a flow of SV 3 to separate the components. In this case, the analytical values of the separated liquids separated into about 5CV each by setting the temperatures of the extract liquid and the column to 20, 40, 50 and 60 ℃ are shown in Table 2.

TABLE 2

Thus, when the separation is performed at a higher temperature, the non-polymer catechins can be more selectively separated and removed.

The variation of the components in the separation at 60 ℃ is shown in FIG. 1.

Example 3

Manufacture of test

600Kg of oolong tea leaves were extracted by column chromatography using 0.15% sodium bicarbonate solution (95 ℃) to obtain about 6000Kg of extract. The liquid temperature was maintained at 60 to 65 ℃ and non-polymeric catechins and caffeine were selectively removed by 400kg of granular activated carbon (GW-H32/60, manufactured by Kuraray). The filtrate was concentrated under reduced pressure to prepare about 900kg of a tea extract having Brix of 10 or more.

The obtained tea extract had 14648ppm of polymerized catechin (OTPP), and 366ppm of adsorbed catechin (EGCG + GCG + ECG + CG).

Example 4

Lipase inhibitory Activity

The extract of example 1 and the tea extract obtained by selectively concentrating (selectively removing non-polymeric catechins) polymeric catechins obtained in example 3 were compared for the inhibitory effect on lipase known to be involved in fat absorption.

Determination of lipase activity:

the lipase activity was measured by measuring the fluorescence of 4-methylumbelliferone produced by the reaction using a fluorescent 4-methylumbelliferone oleate (4-UMO) as a substrate.

For the assay, 13mM Tris-HCl (pH 8.0) containing 150mM NaCl, 1.36mM CaCl2 was used as the buffer. Substrate 4-UMO (manufactured by Sigma Co.) which was a product obtained by diluting a 0.1M DMSO solution prepared 1000-fold with the above buffer, and lipase which was a product obtained by preparing a 400U/ml solution of porcine pancreatic lipase (manufactured by Sigma Co.) with the same buffer were used for enzyme assay.

Enzyme reaction at 25 ℃ in 96 hole enzyme labeling instrument adding and mixing 50 u l 4-UMO buffer solution, 25 u l distilled water (or sample water solution), then by adding 25 u l lipase buffer solution to the reaction start. After the reaction was carried out for 30 minutes, 100. mu.l of 0.1M citric acid buffer (pH 4.2) was added to stop the reaction, and the fluorescence (excitation wavelength 355nm, emission wavelength 460nm) of 4-methylumbelliferone produced by the reaction was measured with a fluorescence analyzer (Fluoroskan asset CF, manufactured by Labsystems).

The inhibitory activity of the test sample was determined as IC50(μ g/ml) in terms of the amount of the sample at which 50% inhibition was observed relative to the activity of the control (distilled water).

The results are summarized in Table 3.

TABLE 3

Example 5

Taste evaluation

The extract of example 1 and the tea extract obtained in example 3 were adjusted to have Brix of 0.5 with distilled water, and sensory evaluation of taste was performed by 11 panelists. The results are summarized in Table 4.

TABLE 4

From the above results, it was confirmed that the tea extract can suppress bitterness while ensuring efficacy.

Example 6

Discussing the ratio by using the effect of inhibiting lipase as an index

The lipase activity of the macromolecular polyphenol fraction produced in preparation example was measured by measuring the fluorescence of 4-methylumbelliferone produced by the reaction using fluorescent 4-methylumbelliferone oleate (4-UMO) as a substrate.

For the assay, 13mM Tris-HCl (pH 8.0) containing 150mM NaCl, 1.36mM CaCl2 was used as the buffer. Substrate 4-UMO (manufactured by Sigma Co.) which was a product obtained by diluting a 0.1M DMSO solution prepared 1000-fold with the above buffer, and lipase which was a product obtained by preparing a 400U/ml solution of porcine pancreatic lipase (manufactured by Sigma Co.) with the same buffer were used for enzyme assay.

Enzyme reaction at 25 ℃ in 96-well microplate reader, 50 u l 4-UMO buffer solution, 25 u l distilled water (or sample water solution) is added and mixed, then by adding 25 u l lipase buffer solution to the reaction start. After the reaction was carried out for 30 minutes, 100. mu.l of 0.1M citric acid buffer (pH 4.2) was added to stop the reaction, and the fluorescence (excitation wavelength 355nm, emission wavelength 460nm) of 4-methylumbelliferone produced by the reaction was measured with a fluorescence analyzer (Fluoroskan asset CF, manufactured by Labsystems).

The inhibitory activity of the test sample was determined as IC50(μ g/ml) in terms of the amount of the sample at which 50% inhibition was observed relative to the activity of the control (distilled water). The reciprocal of the IC50 value was calculated as a relative activity value (unit) per μ g, and the intensity of inhibiting lipase activity in the sample solution was compared.

The beverage was prepared by adding the tea extract (E) obtained in example 3 to oolong tea (OT; Brix0.275) in a concentration of 2 times that of oolong tea (OT; Brix0.275), wherein OT + E was obtained for tea added to Brix0.55, and OT +0.5E was obtained for tea added to tea extract at a concentration of half of that. The oolong tea herein refers to a product obtained by extracting oolong tea leaves with hot water.

TABLE 5

Example 7

Taste evaluation (beverage)

The sensory evaluation of taste was performed by 24 judges on polyphenol-enriched oolong tea (OT + E and OT +0.5E) and usual oolong tea (2OT) at 2-fold concentration, respectively. The results are shown below.

TABLE 6

Based on the above results, when the ratio (weight ratio of solid components) of Oolong Tea (OT) to tea extract (E) is at least 1: 1 to 2: 1, a beverage with reduced bitterness and efficacy can be obtained.

Example 8

Evaluation test of beverage action on human body (verification of neutral fat lowering Effect after eating)

The cross test was performed with healthy adult male and female as test subjects. Each test was carried out with 7 days as a 1 cycle, of which 6 days were a washing period and 7 days were a fat load test. Trial 1 was performed with the oolong tea polyphenol component plus water (2E), trial 2 with polyphenol fortified oolong tea (OT + E), trial 3 with Oolong Tea (OT) and polyphenol fortified oolong tea (OT + 0.5E). 245ml of each was used. The fat-loaded meals were ice cream (2) and egg roll cake (1.5) with a fat intake of 40 g. Blood was collected from the upper arm vein at 0, 1, 3, 4, and 6 hours after the meal load, and the effect of the change in the amount of change in the neutral fat value (Δ TG) in blood after the fat load and the area under the curve (AUC) were compared to confirm the effect of suppressing the increase in the neutral fat value in blood after the fat load in the beverage of 2E, OT +0.5E, OT + E. From this result, it was confirmed that the polymerized catechin can have an effect when fortified to 360mg/L (OT +0.5E) or more than 360mg/L (OT + 0.5E).

The results are shown in FIGS. 2 to 4.

Example 9

Evaluation test of action of beverage on human body (verification of fat excretion promoting Effect)

The cross-over test was performed with healthy adult male female 12 as the test subject. The test beverage used polyphenol fortified oolong tea (OT +0.7E) and the control beverage used water colored with caramel and added with flavors. Each test was a continuous intake of test beverage samples for 10 days, and the last 3 days to compare the total fat excreted in the feces. The washing period was 7 days. As a result, it was confirmed that the polyphenol-enriched oolong tea has a significant fat excretion promoting effect.

The results are shown in FIG. 5.

Since the composition or tea extract of the present invention has no non-polymeric catechin removed, bitterness and astringency are significantly reduced as compared to before the removal of the non-polymeric catechin. Also, caffeine is sufficiently removed by the adsorbent treatment. Therefore, it can be added into tea (tea extract) such as oolong tea and green tea for improving taste.

Also, according to the present invention, it can be demonstrated that the lipase activity inhibitor, composition or tea extract of the present invention has a significant effect of inhibiting lipase activity. Therefore, it can be added to a food or drink, which may be oolong tea, as an additive for suppressing the absorption of fat in the diet, suppressing the increase of neutral fat in the blood, and/or preventing obesity. The composition or tea extract of the present invention is highly safe because it is derived from natural substances, and can be taken and acted on regularly for a long period of time (for example, every several days, every day, or every meal).

The present invention further relates to a pharmaceutical agent for inhibiting dietary fat absorption, inhibiting an increase in neutral fat in blood, and/or preventing obesity, which contains the composition or tea extract of the present invention as an active ingredient for inhibiting lipase. The pharmaceutical product is preferably in a form suitable for oral administration, and may be, for example, in the form of powder, powder or granule, or tablet, pill, oral contraceptive, capsule, troche, candy or chocolate which can be taken directly or dissolved in water for drinking. The amount of the composition or tea extract of the present invention in the pharmaceutical preparation is, for example, 67 to 5000mg of polymerized catechin per ingestion.

Claims (33)

1. A tea extract in which non-polymeric catechins are selectively removed, wherein the content ratio of polymeric catechins to non-polymeric catechins is increased as compared with the original aqueous liquid of an aqueous tea extract.

2. The tea extract according to claim 1, wherein the amount of the polymeric catechin is at least 4 times as large as the non-polymeric catechin.

3. The tea extract according to claim 1 or 2, which is an oolong tea extract.

4. The tea extract according to any one of claims 1 to 3, which is an aqueous liquid component, a composition in a concentrated or dried state.

5. A lipase activity inhibitor comprising the tea extract according to any one of claims 1 to 4.

6. The lipase activity inhibitor according to claim 5, wherein the lipase activity inhibition means inhibition of absorption of fat in the diet, inhibition of increase of neutral fat in blood, and/or prevention of obesity.

7. A process for producing a tea extract as claimed in any one of claims 1 to 4, which comprises bringing an aqueous liquid into contact with an adsorbent to selectively remove non-polymeric catechins, thereby obtaining an aqueous tea extract having an increased content ratio of polymeric catechins to non-polymeric catechins as compared with the aqueous liquid.

8. The method of claim 7, wherein the native aqueous liquid is obtained by: under the slightly alkaline condition, the tea leaves are treated by water with the temperature of 50-99 ℃.

9. The method of claim 8 wherein said water temperature is 80 ℃ to 99 ℃.

10. The method of claim 8, wherein said slight alkalinity is achieved by the addition of sodium bicarbonate.

11. The method of claim 8, wherein the slightly basic is a pH of 8.0 to 8.5.

12. The method of claim 11, wherein said slight alkalinity is a pH of 8.2.

13. The method of claim 7, wherein the concentration of the original aqueous liquid contacted with the adsorbent is Brix 2.0 to 6.0.

14. The method of claim 13, wherein the original aqueous liquor contacted with the adsorbent has a concentration of Brix 3.7.

15. The method of claim 7, wherein the native aqueous liquid is contacted with the adsorbent at a liquid temperature of 50 ℃ or greater than 50 ℃.

16. The process according to claim 15, wherein the adsorption treatment is carried out at a temperature of 50 ℃ to 65 ℃.

17. The method of claim 7, wherein the adsorbent is a selected adsorbent from activated carbon and adsorbent resin.

18. The method of claim 17, wherein the adsorbent is granular activated carbon or synthetic adsorbent resin using polystyrene as a raw material.

19. The process as in claim 18, wherein the granular activated carbon is sized from 32 mesh to 60 mesh.

20. A process according to any one of claims 7 to 19, which comprises packing a chromatographic column with an adsorbent selected from activated carbon or an adsorbent resin, passing the original aqueous liquid through the column, recovering the effluent from the column to obtain a liquid component, and optionally concentrating or drying the effluent to obtain a concentrated product or a dried product.

21. The method of claim 20, wherein the original aqueous liquid having a column capacity of 5 to 10 times that of the original aqueous liquid is passed through the chromatography column.

22. The method of claim 20, wherein the adsorption treatment is performed at a temperature of 50 ℃ or above 50 ℃.

23. The process according to claim 22, wherein the adsorption treatment is carried out at a temperature of 50 ℃ to 65 ℃.

24. The method of claim 20, wherein said adsorbent is granular activated carbon or synthetic adsorbent resin using polystyrene as a raw material.

25. The process as in claim 24, wherein the granular activated carbon is sized from 32 mesh to 60 mesh.

26. An additive for food and drink containing the tea extract according to any one of claims 1 to 4 or the lipase activity inhibitor according to claim 5 or 6.

27. The additive for foods and beverages according to claim 26, which is added to foods and beverages to inhibit absorption of fat in the diet, inhibit increase of neutral fat in the blood, and/or prevent obesity.

28. A food or drink containing the tea extract according to any one of claims 1 to 4 or the lipase activity inhibitor according to claim 5 or 6.

29. The food and drink according to claim 28, which is a health food or health drink.

30. The beverage product according to claim 28 which is a tea beverage.

31. The beverage food product of claim 30 which is an oolong tea beverage.

32. The food or beverage according to any one of claims 28 to 31, wherein the polymerized catechin in the food or beverage is enriched so that the content of polymerized catechin in 1L of the beverage is 268mg or more.

33. The food or beverage according to claim 32, wherein the polymerized catechin in the food or beverage is enriched so that the content of polymerized catechin in 1L of the beverage is 268 to 3600 mg.