TW201332452A - A beverage including coffee extract and the manufacturing method thereof - Google Patents

Abstract

A beverage including coffee extract, comprises: a coffee extract, which includes active element to inhibit fat accumulating; and a substrate, which includes at least one storage stabilizer, the storage stabilizer is selected from a group of sucrose, fructose, fructooligosaccharide (FOS), and isomaltooligosaccharide (IMO). The present invention also discloses a manufacturing method of the beverage, which comprises: a drying step, drying at least a coffee to the water content under 10%, and obtain a dry coffee; a extraction step, adding a solvent to extract active element which can inhibit fat accumulating in the dry coffee, and obtain a coffee extract liquid; a condensation step, removing the solvent and obtaining a coffee extract; and a mixing step, adding the coffee extract into a substrate, and obtain a beverage including coffee extract.

Description

Drink containing raw coffee extract and preparation method thereof

The present invention relates to a beverage containing a coffee extract and a preparation method thereof, and particularly to a beverage containing a green coffee extract for improving or inhibiting fat accumulation, which has better storage stability and preparation of the beverage The method is capable of obtaining a higher concentration of green coffee extract.

In daily life, modern people, in addition to supplementing water to break down thirst, want to be able to meet their needs from a wide variety of drinks. For example, by drinking a beverage containing lactic acid bacteria to adjust the body, improve immunity, or inhibit Helicobacter pylori, choose a tea containing catechin to suppress the rise of body fat, or to enhance the spirit by drinking coffee. Wait.

Many studies have confirmed that green coffee beans contain many physiologically active ingredients. Raw coffee beans contain about 0.6% of coffee sterols (Cafestol), about 0.3% of coffee white fat (Kahweol), and 1.5 to 3.0% of caffeine. (Caffeine) and 4.0 to 8.0% of chlorogenic acid, wherein chlorogenic acid is a phenolic compound having a molecular formula of C ₁₆ H ₁₈ O ₉ and its structure is a cinnamic acid compound such as caffeic acid. ), ferulic acid (ferulic acid) and p-coumaric acid (p -coumaric acid) and other compounds, bonded to an ester bond with quinic acid (quinic acid) (Fujioka et al., as published in Food Chemistry 2008 The journal's paper) has good antioxidant power and can inhibit fat accumulation and regulate the metabolism of sugar.

Previous studies have found that chlorogenic acid has the effect of inhibiting fat accumulation, which is derived from: (1) inhibition of pancreatic lipase activity and reduction of intestinal fat decomposition and absorption (Nazawa et al ., 2009) (2) inhibiting the activity of hepatic fatty acid synthase and reducing fatty acid production (Tanaka et al ., 2009); (3) promoting fatty acid β-oxidation and accelerating fat metabolism (Cho et al) , 2010) (4) inhibiting the differentiation of 3T3-L1 preadipocytes and reducing body fat accumulation (Hsu et al ., 2006).

However, it has not yet been prepared as a functional drink by the active ingredient contained in coffee, which inhibits the accumulation of fat, because the active ingredients such as chlorogenic acid, coffee white fat and coffee sterol contained in the coffee are preserved for a period of time ( Such as one week to one month, may be damaged due to poor storage conditions, or its own structural instability and self-decomposition, the inhibition of fat accumulation of these active ingredients is invalid, so that the drinker after the drink, the active ingredients The concentration is also lowered, so that the drinker cannot obtain a better effect of suppressing fat accumulation by the drink.

Therefore, it is necessary to provide a beverage having a better storage stability of green coffee extract and a preparation method thereof, which enables the beverage to maintain a high concentration after being stored for a long period of time (for example, more than one month). The active ingredient, and enables the drinker to achieve a physiological effect of effectively inhibiting fat accumulation by the high concentration of the active ingredient in the drink.

The main object of the present invention is to provide a beverage containing a green coffee extract which has better storage stability.

A second object of the present invention is to provide a method for preparing a beverage containing a green coffee extract which is effective for enhancing an active ingredient which effectively inhibits fat accumulation in the beverage.

In order to achieve the foregoing object, the technical content of the present invention includes:

A beverage containing a raw coffee extract, comprising: a raw coffee extract, comprising an active ingredient for inhibiting fat accumulation, the active ingredient comprising chlorogenic acid; and a substrate solution comprising at least one storage stabilizer, the storage The stabilizer is selected from the group consisting of sucrose, fructose, fructooligosaccharide and isomalt oligosaccharide, and the storage stabilizer has a concentration by weight of 5 to 15%.

The present invention contains a green coffee extract containing 5 mg/mL of green coffee extract.

In the beverage containing the green coffee extract of the present invention, the pH value of the beverage is preferably pH 7-8.

In the beverage containing the green coffee extract of the present invention, the beverage is preferably additionally added with an antioxidant, and the antioxidant may be selected from ascorbic acid.

In the beverage containing the green coffee extract of the present invention, the beverage is preferably additionally added with an inorganic salt selected from the group consisting of sodium chloride, calcium chloride, potassium chloride and magnesium chloride.

The method for preparing a beverage containing a green coffee extract comprises: a drying step of drying at least one raw coffee to a moisture content of less than 10% to obtain a dried green coffee; and an extraction step of methanol and ethanol Or ethyl acetate as an extraction solvent, extracting the active ingredient in the dried green coffee for inhibiting fat accumulation, to obtain a raw coffee extract; and a concentration step, removing the solvent of the green coffee extract to obtain a raw coffee extract; And a mixing step of adding the raw coffee extract to a substrate solution, the matrix solution comprising at least one storage stabilizer selected from the group consisting of sucrose, fructose, fructooligosaccharide and isomalt oligosaccharide In the group, the substrate solution contains a storage stabilizer in a concentration of 5 to 15% by weight.

In the preparation method of the beverage containing the green coffee extract of the present invention, the green coffee extract is preferably obtained by extracting unbaked green coffee fruit, green coffee beans or a combination thereof.

In the preparation method of the beverage containing the green coffee extract of the present invention, the unroasted green coffee fruit and green coffee beans are preferably selected from the 7th to 10th month after flowering.

In the preparation method of the beverage containing the green coffee extract of the present invention, the extraction time of the extraction step is preferably 2 to 4 hours.

In the preparation method of the beverage containing the green coffee extract of the present invention, the pH value of the beverage is preferably pH 7-8.

In the preparation method of the beverage containing the green coffee extract of the present invention, the matrix solution is preferably additionally added with an antioxidant, and the antioxidant may be selected as ascorbic acid.

In the preparation method of the beverage containing the green coffee extract of the present invention, the beverage is further added with an inorganic salt selected from the group consisting of sodium chloride, calcium chloride, potassium chloride and magnesium chloride.

The above and other objects, features and advantages of the present invention will become more <RTIgt;

The beverage containing the raw coffee extract of the present invention comprises a primary coffee extract, comprising an active ingredient for inhibiting fat accumulation, the active ingredient comprising chlorogenic acid; and a substrate solution comprising at least one storage stabilizer. The storage stabilizer is selected from the group consisting of sucrose, fructose, fructooligosaccharide and isomalt oligosaccharide, so that the beverage containing the green coffee extract can be stored at a temperature of -20 ° C to 55 ° C. For up to three months, the active ingredient used to inhibit fat accumulation can still reach more than 50%. The pH value of the beverage containing the green coffee extract is preferably pH 7-8.

The raw coffee of the present embodiment refers to coffee fruit, coffee beans and combinations thereof from three major stocks, wherein the three major species include but are not limited to Coffea arabica or Robusta coffee ( Coffa Robusta) Libyan and coffee (Coffea liverica) and the like, wherein the coffee cherries, coffee bean, and combinations thereof, preferably selected after flowering lines 7 to 10 months in order to inhibit fat accumulation and extracted several active ingredients, such as chlorogenic acid And caffeine, wherein the chlorogenic acid includes: 3-caffeoylquinic acid, 3-CQA; 4-caffeoylquinic acid, 4-CQA or 5-caffeoylquinic acid, 5-CQA, 3,4-dicaffeoylquinic acid, 3,4-diCQA; 4,5-dicaffeoylquinic acid, 4,5-CQA or 3,5-dicaffeoylquinic acid, 3,5-CQA) and avidinic acid (3- A compound such as feruloylquinic acid, 3-FQA, 4-feruloylquinic acid, 4-FQA or 5-feruloylquinic acid (5-FQA), and a plurality of active ingredients for inhibiting fat accumulation, hereinafter referred to as active ingredients. This example selects, but does not limit, the Arabica coffee grown in Dongshan Township, Tainan County, to prepare the beverage and conduct the following tests.

Referring to FIG. 1, the beverage containing the green coffee extract of the present invention is prepared by the following steps: a drying step S1, an extraction step S2, a concentration step S3, and a mixing step S4.

The drying step S1 is to dry at least one raw coffee to a moisture content of less than 10% to obtain a dried green coffee. More specifically, in this embodiment, the unroasted green coffee fruit, green coffee beans or a combination thereof is dried to reduce the water activity of the green coffee fruit, green coffee beans or a combination thereof, thereby avoiding The active ingredient contained in the fat-inhibiting decomposition is decomposed or self-degraded, and helps to retain the high content of the active ingredient in the raw coffee or green coffee beans.

In the extraction step S2, methanol, ethanol or ethyl acetate is used as an extraction solvent, and an active ingredient for suppressing fat accumulation in the dried green coffee is extracted to obtain a primary coffee extract. This embodiment is selected, but not limited to, to grind the dried green coffee to a screen that can pass through a mesh of 60 mesh (i.e., particles or powders having a diameter of 1.5 mm or less) to help improve the extraction efficiency of the active ingredient. More specifically, the dried green coffee is preferably ground to a suitable size of coffee granules or powder, and the active ingredient is extracted by a cable extractor; for example, the present embodiment is selected from methanol, ethanol and acetic acid. One of the ethyl esters is used as an extraction solvent, and after extracting at a temperature of 80 ° C for 2 to 4 hours, the green coffee extract is collected, and more preferably extracted with methanol or ethanol at a concentration of 70 to 100%. In this embodiment, the extraction is carried out after 7 to 10 months of raw coffee fruit or coffee beans, which has a high chlorogenic acid content.

The concentration step S3 removes the solvent of the green coffee extract to obtain a raw coffee extract. More specifically, the present embodiment is used for extracting the dried green coffee, and contains an active ingredient for inhibiting fat accumulation, mainly a compound such as chlorogenic acid and caffeine. The coffee extract is concentrated and dried to obtain the green coffee extract by means of concentration under reduced pressure or freeze-drying, and the green coffee extract is in the form of a powder, so that the raw coffee extract can be subsequently added to the drink, and the raw coffee can be conveniently adjusted. The concentration of the coffee extract in the drink.

In the mixing step S4, the green coffee extract is added to a substrate solution to obtain a beverage containing a green coffee extract, wherein the matrix solution comprises at least one storage stabilizer selected from the group consisting of sucrose, fructose, and fructooligosaccharide. And the group consisting of isomalto-oligosaccharides, the substrate solution contains a storage stabilizer at a concentration of 5 to 15% by weight; and the storage stabilizer of the embodiment is selected to be 10%. For example, the substrate solution can be any beverage with good taste, such as water, coffee, juice, fruit tea or herbal tea, and the beverage can be added with other additives, such as antioxidants (such as EDTA). , perfumes, organic acids, organic acid salts, inorganic acids, inorganic acid salts, pigments, emulsifiers, flavoring agents, sweeteners, vitamins, amino acids, fruit juice extracts, vegetable extracts or nectar extracts, etc. . Wherein, the fragrance, the coloring matter, the flavoring agent, the sweetener, the juice extract, the vegetable extract or the nectar extract, etc., can be used to improve the taste of the drink, and the antioxidant is used to stabilize the active ingredient; The vitamins include vitamins A, B, C, D or E to increase the nutritional value of the drink.

In order to confirm that the beverage containing the green coffee extract of the present invention can maintain its effect of inhibiting fat accumulation under a long period of time (at least three months), the following test is carried out: (A) extraction solvent test, B) preferred extraction concentration test for different extraction solvents, (C) preferred extraction time test, (D) green coffee test of different maturity, (E) storage temperature test of the beverage containing the green coffee extract of the present invention, ( F) a storage pH test of the beverage containing the green coffee extract of the present invention and (G) a preferred storage stabilizer test of the beverage containing the green coffee extract of the present invention, and confirming that the present invention contains a raw animal in a hyperlipidemia mode Drinks from coffee extracts are indeed effective in inhibiting fat accumulation (Test H).

The test results of the tests (A) to (G) of the present embodiment include measuring the chlorogenic acid content in the green coffee extract or drink, and also performing the pancreatic lipase of the green coffee extract or drink. An activity test to confirm the effect of different embodiments on inhibiting fat accumulation, wherein the pancreatic lipolytic enzyme adheres to the fat emulsified particles for hydrolysis, and the triglyceride is decomposed into monoglycerides (mono Glyceride and free fatty acid make small intestinal cells easy to absorb, and then fat cells in the body re-synthesize fat or sterols. Therefore, the higher the activity of the pancreatic lipolytic enzyme, the more fat that the organism can absorb, resulting in fat accumulation.

The lipolytic enzyme reaction with a substrate (p -nitrophenyl palmitate, abbreviated p NPP): In this embodiment pancreatic lipolytic activity is determined based on the reference method McDougall et al, Journal of Food Chemistry 2009, which works as follows Analysis , which is decomposed into p- nitrophenol ( p NP) and Palmitic acid. The p NP has a maximum absorbance at a wavelength of 400 nm. The absorbance (AB) of the control group and the experimental group are measured to convert the test group fat. The percentage of activity of the degrading enzyme (hereinafter referred to as the lipolytic enzyme inhibition rate), wherein the lipolytic enzyme inhibition rate is calculated as follows:

More specifically, in the present embodiment, after preparing the test solution as shown in Table 1, it is preferred to use 100 μL of the beverage containing the green coffee extract and 100 μL of the pancreatic lipolytic enzyme solution (2.5 mg/ mL, purchased from Sigma, USA, after reacting at 37 ° C for 10 minutes, adding 600 μL of the third solution to react at 37 ° C for 30 minutes, and then adding 500 μL of trichloroacetic acid solution to terminate the reaction, to 5570 × After centrifugation for 10 minutes, 500 μL of the supernatant was added, and 300 μL of sodium hydroxide solution (1 M, purchased from Nacalai Tesque, Japan) was added, and the absorbance at a wavelength of 400 nm was measured with a spectrophotometer. Phosphate buffer solution without p NPP was used as a blank group (Blank).

In addition, in this example, the measurement of chlorogenic acid was carried out by high performance liquid chromatography (HPLC). For example, in this embodiment, the green coffee extract is formulated into a solution of 1000 μg/mL in 70% methanol, filtered through a 0.45 μm filter, and used as a sample solution for HPLC quantification; and a chlorogenic acid is further prepared. Standards (concentrations of 100, 200, 300, 400, 500, 600 μg/mL, purchased from Sigma, USA), filtered through a 0.45 μm filter, and injected into HPLC for analysis, with the standard wave front area as the Y axis And the standard concentration is the X axis, draw a standard curve, and find the linear regression equation (y = ax + b) and the coefficient of determination (r ² ).

The HPLC analysis conditions of this example were as follows: the column was Agilent ZORBAX SB-C18 column (4.6 x 150 mm, 5 μm), the column temperature was 30 ° C, the mobile phase was Acetonitrile-0.4% phosphoric acid, and the flow rate was set to 1 mL/min, each injection volume is 20 μL, and the absorbance at a wavelength of 318 nm is measured, and the measured absorbance is brought into the linear regression equation of the standard curve to obtain the content of the chlorogenic acid.

(A) Extraction solvent test

The beverage containing the green coffee extract of the present invention is obtained by using methanol or ethanol as an extraction solvent, and the obtained green coffee extract contains a high active ingredient for inhibiting fat accumulation. For example, in this embodiment, five sets of dried green coffee (100 grams each) are taken, respectively, with 40 volumes of methanol (purchased from Taiwan Jingming Chemical Co., Ltd.) and ethanol (purchased from Taiwan Province tobacco and alcohol company). ), ethyl acetate (purchased from Taiwan Jingming Chemical Co., Ltd.), acetone (purchased from Taiwan Jingming Chemical Co., Ltd.) and water as extraction solvent, extracted with a cable extractor for 2 hours, and collected the green coffee extract, and The green coffee extract was dried to obtain a green coffee extract, and the active ingredient content of each group of raw coffee extracts (represented by the content of chlorogenic acid in this example) and its activity of inhibiting lipolytic enzyme were measured.

Please refer to Table 2 and Figures 2 and 3 for the extraction conditions, the chlorogenic acid content and the lipolytic enzyme inhibition rate obtained under the extraction conditions of the examples.

As can be seen from Figures 2 and 3, in the present example, the A2 group extracted with ethanol has more chlorogenic acid (300 μg/mg or more), and the lipolytic enzyme inhibition rate can reach 70% or more, followed by methanol. The extracted Group A1 and the A3 group of ethyl acetate all contain at least 250 μg/mg of chlorogenic acid, and the lipolytic enzyme inhibition rate can reach 50% or more.

It can be seen that the beverage containing the green coffee extract of the present invention can be extracted by using methanol, ethanol or ethyl acetate as an extraction solvent to obtain a higher content of the active ingredient, and the active ingredient can effectively inhibit the lipolytic enzyme. The activity has the effect of providing a beverage that is effective in inhibiting fat accumulation.

(B) Preferred extraction concentration test for different extraction solvents

The beverage containing the green coffee extract of the invention is prepared by using 60-100% methanol or ethanol as an extraction solvent, and the obtained raw coffee extract contains a high active ingredient for inhibiting fat accumulation. For example, in this embodiment, 7 sets of dried green coffee (100 grams each) are taken, and 40 times volume of different concentrations of methanol and ethanol are used as extraction solvents, and extracted by a cable extractor for 2 hours, and collected. The raw coffee extract is dried, and the raw coffee extract is dried to obtain a green coffee extract, and the active ingredient content of each group of raw coffee extracts is measured (this example is represented by the content of chlorogenic acid) and its inhibition The activity of lipolytic enzymes.

Please refer to Table 3 and Figures 4 and 5 for the extraction conditions, the chlorogenic acid content and the lipolytic enzyme inhibition rate of each extraction condition.

As can be seen from Figures 4 and 5, in Group B7, which was extracted with 95% ethanol in this example, had more chlorogenic acid (300 μg/mg or more), and its lipolytic enzyme inhibition rate was over 70%, followed by Extracted from 70% ethanol, 80% methanol, 100% methanol and 50% ethanol, all contain at least 250 μg/mg of chlorogenic acid, and the lipolytic enzyme inhibition rate can reach more than 50%.

It can be seen that the beverage containing the green coffee extract of the present invention can be extracted by 70-100% methanol or ethanol, and can extract a high content of the active ingredient, and the active ingredient inhibiting fat accumulation can be effectively effective. Inhibiting the activity of lipolytic enzymes provides a method of extracting active ingredients effective to inhibit fat accumulation at a lower concentration of solvent to reduce the cost of preparation of the beverage.

(C) preferred extraction time test

The beverage containing the raw coffee extract of the invention adopts 95% ethanol as an extraction solvent, and after the extraction time is 2 to 4 hours, the obtained green coffee extract contains a high effective for inhibiting fat accumulation. ingredient. For example, in this embodiment, four sets of dried green coffee (100 grams each) are taken, and 40 times volume of 95% ethanol is used as an extraction solvent, and extracted by a cable extractor for 2 to 4 hours, and collected. The raw coffee extract is dried, and the green coffee extract is dried to obtain a green coffee extract, and the active ingredient content of each group of raw coffee extracts is measured (this example is represented by the content of chlorogenic acid) and its inhibition The activity of lipolytic enzymes.

Please refer to Table 4 and Figures 6 and 7 for the extraction conditions, the chlorogenic acid content and the lipolytic enzyme inhibition rate of each extraction condition.

As can be seen from Figures 6 and 7, in the present embodiment, the C2 group having an extraction time of 2 hours has more chlorogenic acid (300 μg/mg or more), and the lipolytic enzyme inhibition rate can reach 70% or more. The next 3 and 4 hour groups contain about 300 μg/mg or more of chlorogenic acid, and the lipolytic enzyme inhibition rate can reach more than 65%.

It can be seen that the beverage containing the green coffee extract of the present invention is extracted with 95% ethanol for 2 to 4 hours, and can be extracted to obtain a higher content of the active ingredient, and the active ingredient can effectively inhibit the activity of the lipolytic enzyme. To provide a method for extracting active ingredients for inhibiting fat accumulation, which is capable of extracting the active ingredients in the raw coffee almost completely, in order to achieve the full utilization of resources.

(D) Raw coffee test with different maturity

The beverage containing the green coffee extract of the present invention is extracted from the coffee fruit and the coffee beans in the 5th to 10th month after flowering, and the obtained green coffee extract contains a higher content of the active ingredient. For example, in this embodiment, 6 sets of dried green coffees of different maturity (100 grams each) are taken, and 40 times volume of 95% ethanol is used as an extraction solvent, and extracted by a cable extractor for 2 hours. Collecting the raw coffee extract, and drying the green coffee extract to obtain a green coffee extract, and measuring the active ingredient content of each group of raw coffee extracts (this embodiment is represented by the content of chlorogenic acid) and It inhibits the activity of lipolytic enzymes.

Please refer to Table 5 and Figures 8 and 9 for the extraction conditions of this example, while Figures 8 and 9 are for the chlorogenic acid content and the lipolytic enzyme inhibition rate of each group. .

It can be seen from Figures 8 and 9 that the chlorogenic acid content of the raw coffee fruit increased significantly in the 5th to 6th month after flowering, and the chlorogenic acid content of the raw coffee fruit after 7th to 10th month after flowering was flat. It does not increase, and has chlorogenic acid of 300 μg/mg or more, and its lipolytic enzyme inhibition rate can reach 50% or more.

It can be seen that the beverage containing the green coffee extract of the present invention is preferably extracted from the raw coffee of the 7th to 10th month after flowering, thereby obtaining a higher content of the active ingredient, and the active ingredient can effectively inhibit the fat. The activity of the enzyme is solved to prepare raw coffee containing a higher content of the active ingredient, so as to improve the effect of suppressing fat accumulation of the beverage containing the green coffee extract.

After extracting the primary coffee from the conditions shown in the tests (A) to (D), a green coffee extract containing a higher content of the active ingredient can be obtained, and the raw coffee extract is prepared into a drink, wherein the drink is It can be stored in an environment with a temperature of -20~55 ° C, and the pH value of the drink is preferably pH 7-8, and the beverage can be additionally added with a storage stabilizer, which is selected from the group consisting of sucrose and fructose. The group consisting of oligosaccharides and isomalto-oligosaccharides can contribute to the long-term storage stability of the active ingredient.

(E) Storage temperature test of the beverage containing the green coffee extract of the present invention

The beverage containing the raw coffee extract of the invention can be stored for at least three months at a temperature of -20 to 55 ° C, and the active ingredient of the raw coffee extract can still retain more than 10% of lipolytic enzyme inhibition. The rate is a kind of drink that can provide an organism that effectively inhibits fat accumulation. In this embodiment, the semi-inhibitory concentration test of the green coffee extract for the lipolytic enzyme is preferably carried out, for example, the raw coffee extracts are respectively formulated into different concentrations (1.5, 2.0, 2.5, 3.0, and 3.5 mg/ (mL) raw coffee extract aqueous solution, and measure the inhibition rate of lipolytic enzymes of each group, the concentration of each group is X axis, the lipolytic enzyme inhibition rate is Y axis, calculate the linear regression equation and its coefficient of determination (r ² ) The linear regression equation was y=24.84x-16.06, r ² =0.9939; the half-inhibitory concentration of the green coffee extract of the present invention was determined to be 2.65 mg/mL.

Please refer to Table 6 and Figures 10 and 11, for the storage type and temperature of each group in this example, and Figures 10 and 11 for the lipolysis of each group after an interval of storage. The enzyme inhibition rate line graph, wherein Fig. 10 is a graph showing the results of the aqueous solution of the green coffee extract, and Fig. 11 is a graph showing the results of the dried powder of the green coffee extract.

It can be seen from Fig. 10 that when the green coffee extract is prepared into an aqueous solution (containing 5 mg/mL of green coffee extract), the storage stability of the dried powder of the green coffee extract is poor, wherein, the storage is poor. At 4 ° C (group E1-2) and room temperature (group E1-3), the inhibition ability of lipolytic enzyme activity decreased rapidly. When stored for two weeks, the inhibition rate of lipolytic enzyme in group E1-2 was 62.6. % decreased to 33.7%, while the E1-3 group decreased from 66.5% to 36.2%. After three months of storage, the lipolytic enzyme inhibition rates of the E1-2 and E1-3 groups decreased to 9.5%, respectively. 10.1%; compared with the E1-2 and E1-3 groups, the inhibition rate of lipolytic enzyme in the E1-1 group (-20 °C) was slower, and the lipid in the E1-1 group was stored four weeks after storage. The inhibition rate of enzyme solution decreased from 68.9% to 41.7%, and decreased to 14.5% after three months. The inhibition rate of lipolytic enzyme in group E1-4 (55 °C) decreased the most, after three months of storage, E1 The lipolytic enzyme inhibition rate of the -1 group was reduced from 66.2% to 40.5%.

It can be seen from Fig. 11 that the lipolysis enzyme inhibition rate of the green coffee extract after storage for three months at different temperatures, at -20 ° C (group E2-1), 4 ° C (group E2-2) At room temperature (Group E2-3) and 55 °C (Group E2-4) for three months, the inhibition rate of lipolytic enzymes in each group can be maintained above 60%, confirming the raw coffee extract Although it has better stability in a dry state, the green coffee extract can be formulated into a form that is easy for the user to drink (e.g., a drink), which still has a physiological effect of inhibiting fat accumulation.

It can be seen that the beverage containing the green coffee extract of the present invention is preferably prepared in a dry form and then prepared into a drink before drinking. The storage stability of the active ingredient in the drink is better, thereby prolonging the activity. The storage time of the component and the activity of the lipolytic enzyme are effectively inhibited to enhance the effect of suppressing fat accumulation of the beverage containing the green coffee extract.

(F) Storage pH test of the beverage containing the green coffee extract of the present invention

The storage pH value of the beverage containing the green coffee extract of the invention is adjusted and stored under the condition of pH 7-8 for at least three months, and the active ingredient can still maintain the inhibition rate of lipolytic enzyme by more than 40%, which can provide Organism A drink that effectively inhibits the accumulation of fat. In this embodiment, the green coffee extract is prepared into an aqueous solution having a concentration of 5 mg/mL in a citrate buffer solution of pH 3-8, and stored for three months at a temperature of 4 °C.

As can be seen from Fig. 12, the pH value of the green coffee extract is adjusted to pH 3, 4, 5, 6, 7, and 8 (in the order of F1 to F6), wherein the storage pH value is pH 3~ 4 (Groups F1 and F2), the inhibition rate of lipolytic enzyme decreased rapidly after two weeks of storage, and after three months of storage, the inhibition rate of lipolytic enzyme decreased to 25.5% and 31%, respectively; The base value was pH 5~6 (groups F3 and F4). Although the inhibition rate of lipolytic enzyme was stable for two weeks after storage, the inhibition rate of lipolytic enzyme in group F3 decreased to 33.2% after three months of storage. In group F4, it decreased to 33.7%; the pH value of storage and storage was pH7~8 (groups F5 and F6), and the inhibition rate of lipolytic enzyme was slower. After three months of storage, the group F5 The lipolytic enzyme inhibition rate decreased from 68.9% to 49.8%, and the F6 group decreased from 66.2% to 41.6%.

It can be seen that the raw coffee extract is stored in a neutral pH environment, which helps to maintain a 50% or more lipolytic enzyme inhibition rate after long-term storage (three months). Although the raw coffee extract has better stability in a dry state, it helps the green coffee extract-containing beverage to retain the effect of inhibiting fat accumulation in the case of long-term storage.

(G) A preferred storage stabilizer test for a beverage containing a green coffee extract of the present invention

The beverage containing the raw coffee extract of the present invention can be stored for at least three months after adding a storage stabilizer, and the raw coffee extract can still retain 30% or more of the lipolytic enzyme inhibition rate, and can provide the living body. A drink that effectively inhibits fat accumulation.

In this embodiment, the green coffee extract is formulated into an aqueous solution having a concentration of 5 mg/mL, and a storage stabilizer is added, and the storage stabilizer may be selected from the group consisting of sucrose, fructose, fructooligosaccharide and isomaltose oligosaccharide. The group is composed and stored for three months at a temperature of -20 to 55 °C. More specifically, the saccharide used in the present embodiment as a storage stabilizer can increase the stability of chlorogenic acid, prevent chlorogenic acid from decomposing during storage, and maintain its lipolytic enzyme activity inhibiting ability.

It can be seen from Fig. 13 that sucrose, fructose, fructooligosaccharide and isomalto-oligosaccharide are added in 10% by weight to the aqueous solution of the raw coffee extract of the G1-1 to G1-4 groups, respectively. For the sugar, another group G1-5 without a storage stabilizer was used as the control group, and the G1-2 and G1-4 groups all had the effect of slowing down the inhibition rate of the lipolytic enzyme compared with the control group. After three months of storage, the inhibition rate of lipolytic enzymes in each group can be higher than 50%; although the remission effect of the G1-1 group is poor, after three months of storage, it can still make it lipolytic The enzyme inhibition rate is maintained to about 30%.

As can be seen from Fig. 14, in the aqueous solution of the raw coffee extract of the G2-1 to G2-4 group, sodium chloride, calcium chloride, potassium chloride and magnesium chloride having a concentration of 5 mM were sequentially added, and another one was set. The G2-5 group without the storage stabilizer was the control group, and the G2-1, G2-2 and G2-4 groups all had the effect of slowing down the inhibition rate of the lipolytic enzyme compared with the control group. After three months of storage, the inhibition rate of lipolytic enzymes in each group was higher than 30%; while the remission effect in the G2-3 group was poor, and the lipolytic enzyme inhibition rate was still obtained after six weeks of storage. Maintained to about 30%, but only 10% of the lipolytic enzyme inhibition rate remained for three months.

As can be seen from Fig. 15, in the aqueous solution of the raw coffee extract of the G3-1 and G3-2 groups, 0.2% by weight of citric acid and 0.05% by weight of ascorbic acid were added, and another The G3-3 group without the storage stabilizer was the control group, and the G3-2 group had the effect of slowing down the inhibition rate of the lipolytic enzyme compared with the control group, and after three months of storage, The lipolytic enzyme inhibition rate was higher than 20%; while the G3-1 group had a poorer alleviation effect, after six weeks of storage, the lipolytic enzyme inhibition rate was maintained to about 30%, but stored for three months. Only 10% of the lipolytic enzyme inhibition rate remains.

It can be seen that the raw coffee extract is additionally added with a storage stabilizer, such as sugars such as sucrose, fructose, fructooligosaccharides and isomalto-oligosaccharides, or metal ions such as sodium ions, potassium ions and magnesium ions or ascorbic acid. Helps to maintain more than 20% lipolytic enzyme inhibition rate after long-term storage (three months), among which sugar is the main storage stabilizer, which really helps the invention to contain green coffee extract The beverage has better storage stability during long-term storage, and the green coffee extract beverage retains the physiological effect of inhibiting fat accumulation after long-term storage.

(H) High blood lipid animal test

In order to confirm that the beverage containing the green coffee extract of the present invention has an effect of effectively inhibiting the accumulation of fat, this embodiment refers to the "Principle I" animal proposed in the "Method for Evaluating the Body Fat Function of Healthy Foods" which is published by the Department of Health of the Executive Yuan. In the test mode, after the obesity phenomenon of the experimental animal is induced by the high-calorie feed, the beverage containing the green coffee extract of the present invention is further administered, and the beverage containing the green coffee extract of the present invention is judged by various indexes to have the effect of inhibiting fat accumulation.

In this example, 36 male Sprague-Dawley rats (eight weeks old, weighing about 200-250 g) purchased from the National Laboratory Animal Breeding and Research Center were selected and randomly divided into four groups. The animals were kept in the normal range of temperature (25±1 °C) and light (12 hours of sunshine), and the rats were given free access to food and water. The feeding conditions of each group are shown in Table 7, which is the H1. In the H4 group, the induction of the hyperlipidemic animal model of the present example was completed after the 12th week of feeding, wherein the body weight of the H2 to H4 group was 13-21% of the H1 group, and the latter five weeks was 34.8. The coffee extract of mg/kg/day was fed to the H3 group, and the H4 group was fed at 174.1 mg/kg/day, and the body weight was measured after six weeks, and the blood and the adipose tissue of the kidney and the testicle were taken. In the analysis, among the blood biochemical index, the serum total cholesterol content of the H3 and H4 groups was about 15~25 mg/dl lower than that of the H2 group, and the triglyceride content was about 30-40 mg/dl lower; There were no significant differences between ester and low density ester proteins.

Please refer to Table 8, which is the percentage of body weight of the groups in this example at the end of the experiment and the weight of adipose tissue located in the kidney and testicles. The body weight of the H2 group was significantly higher than that of the H3 and H4 groups, and the weight of the adipose tissue of the H3 and H4 groups was also lower than that of the H2 group, indicating that the beverage of the green coffee extract of the present invention can actually reduce body fat accumulation.

It can be seen from the above tests (E) to (H) that the beverage containing the green coffee extract of the present invention is composed of at least one storage stabilizer, and is selected from the group consisting of sucrose, fructose, fructooligosaccharide and isomalt oligosaccharide. Group, or add an inorganic salt (such as sodium, potassium and magnesium) or an antioxidant (such as ascorbic acid) to enhance the storage stability of the active ingredient in the drink to inhibit fat accumulation, and the drink does It has the effect of inhibiting the accumulation of fat in the body.

In the beverage containing the raw coffee extract of the present invention, the active ingredient for inhibiting fat accumulation is mainly chlorogenic acid derived from the raw coffee extract, preferably before the user drinks, and the raw coffee extract is dissolved in the raw coffee extract. a solution containing a storage stabilizer to prepare a beverage containing the green coffee extract for consumption by a user; or storing the beverage of the present invention at a temperature of -20 ° C to 55 ° C, or an acid or alkali of the beverage The value is adjusted to pH 7~8. Thus, the beverage containing the green coffee extract of the present invention can withstand long-term storage (for example, three months or more), and the active ingredient contained therein for suppressing fat accumulation can still maintain 30~60. The inhibition rate of %, therefore, the beverage containing the green coffee extract of the present invention can effectively inhibit the accumulation of fat in the living body, and can further be used to control the body weight of the organism.

The beverage containing the green coffee extract of the present invention retains the effect of suppressing fat accumulation by 50% or more after the long-term storage of the beverage by using a storage stabilizer. The storage of the active ingredient in the beverage is periodically extended.

The method for preparing a green coffee extract-containing beverage of the present invention, which comprises an active ingredient which effectively inhibits fat accumulation in the beverage, retains a 50% inhibition rate after long-term storage, so that the drinker can Drink the drink to achieve effective inhibition of fat accumulation.

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

S1. . . Drying step

S2. . . Extraction step

S3. . . Concentration step

S4. . . Mixing step

Fig. 1 is a block diagram showing the steps of a method for preparing a beverage containing a green coffee extract of the present invention.

Figure 2: Bar graph of chlorogenic acid content in groups A1 to A5 of this example.

Fig. 3 is a bar graph of the lipolytic enzyme inhibition rate of the groups A1 to A5 of the present Example.

Figure 4: Bar graph of chlorogenic acid content in groups B1 to B7 of this example.

Fig. 5 is a bar graph of the lipolytic enzyme inhibition rate of the groups B1 to B7 of the present Example.

Figure 6: Bar graph of chlorogenic acid content in groups C1 to C4 of this example.

Fig. 7 is a bar graph of the lipolytic enzyme inhibition rate in the groups C1 to C4 of the present Example.

Figure 8: Bar graph of chlorogenic acid content in groups D1 to D6 of this example.

Fig. 9 is a bar graph of the lipolytic enzyme inhibition rate in the groups D1 to D6 of the present Example.

Fig. 10 is a line diagram showing the rate of inhibition of lipolytic enzymes in the groups E1-1 to E1-4 of the present Example.

Fig. 11 is a line diagram showing the rate of inhibition of lipolytic enzymes in the groups E2-1 to E2-4 of the present Example.

Fig. 12 is a line diagram showing the rate of inhibition of lipolytic enzymes in the groups F1 to F6 of the present Example.

Fig. 13 is a line diagram showing the rate of inhibition of lipolytic enzymes in the G1-1 to G1-5 groups of the present Example.

Fig. 14 is a line diagram showing the rate of inhibition of lipolytic enzymes in the groups G2-1 to G2-5 of the present Example.

Fig. 15 is a line diagram showing the rate of inhibition of lipolytic enzymes in the G3-1 to G3-3 groups of the present Example.

S1. . . Drying step

S2. . . Extraction step

S3. . . Concentration step

S4. . . Mixing step

Claims (14)

A beverage containing a raw coffee extract, comprising: a raw coffee extract, comprising an active ingredient for inhibiting fat accumulation, the active ingredient comprising chlorogenic acid; and a substrate solution comprising at least one storage stabilizer, the storage The stabilizer is selected from the group consisting of sucrose, fructose, fructooligosaccharide and isomalt oligosaccharide, and the storage stabilizer has a concentration by weight of 5 to 15%. A green coffee extract-containing beverage according to claim 1, wherein the drink contains 5 mg/mL of green coffee extract. A beverage containing a green coffee extract according to claim 1 of the patent application, wherein the beverage has a pH of pH 7-8. A drink containing a green coffee extract according to claim 1 of the patent application, wherein the drink is additionally added with an antioxidant. A beverage containing a green coffee extract according to claim 4, wherein the antioxidant is ascorbic acid. The beverage containing the green coffee extract according to claim 1, wherein the beverage is further added with an inorganic salt selected from the group consisting of sodium chloride, calcium chloride, potassium chloride and magnesium chloride. The group that makes up. A method for preparing a beverage containing a green coffee extract comprises: a drying step of drying at least one raw coffee to a moisture content of less than 10% to obtain a dried green coffee; and an extraction step of methanol or ethanol or Ethyl acetate as an extraction solvent, extracting the active ingredient in the dried green coffee for inhibiting fat accumulation, and obtaining a raw coffee extract; and a concentration step of removing the solvent of the green coffee extract to obtain a raw coffee extract; a mixing step of adding the green coffee extract to a substrate solution comprising at least one storage stabilizer selected from the group consisting of sucrose, fructose, fructooligosaccharides and isomaltose oligosaccharides In the group, the substrate solution contains a storage stabilizer in a concentration of 5 to 15% by weight. The method for preparing a green coffee extract-containing beverage according to claim 7, wherein the green coffee extract is obtained by extracting unbaked green coffee fruit, green coffee beans or a combination thereof. The method for preparing a beverage containing green coffee extract according to claim 8 of the patent application, wherein the unroasted green coffee fruit and green coffee beans refer to the 7th to 10th months after flowering. The method for preparing a beverage containing green coffee extract according to claim 7 of the patent application, wherein the extraction step is performed for 2 to 4 hours. The method for preparing a beverage containing green coffee extract according to claim 7 of the patent application, wherein the drink has a pH of pH 7-8. The method for preparing a beverage containing green coffee extract according to claim 7 of the patent application, wherein the substrate solution is further added with an antioxidant. A method for preparing a beverage containing a green coffee extract according to claim 12, wherein the antioxidant is ascorbic acid. The beverage containing green coffee extract according to claim 7 of the patent application, wherein the beverage is further added with an inorganic salt selected from the group consisting of sodium chloride, calcium chloride, potassium chloride and magnesium chloride. The group that makes up.