JP5886367B2 - Deodorant composition - Google Patents

Description

  The present invention relates to a deodorizing composition containing laccase and mate tea extract, and a food or drink containing the same.

Conventionally, as a method for deodorizing and deodorizing malodors, a method of removing malodors by masking with an aromatic substance, chemical deodorization with an oxidizing agent, neutralizing agent, fixing agent, etc., or adsorption with activated carbon or the like has been used. It was. However, these methods often have significant restrictions depending on the purpose of use. Particularly in the food field, it is necessary to use substances that are safe for the human body when ingested.
Natural product extracts such as herbal medicines and herbs that have been used since ancient times have attracted attention as deodorants with excellent deodorizing effects that have solved these problems, and have been used since ancient times. It is known that there is an effect (patent documents 1-7, non-patent documents 1-3). In particular, Non-Patent Document 2 describes a deodorizing effect by using a combination of chlorogenic acid, which is the main component of mate tea, and an apple extract having a polyphenol oxidase effect.

  However, it has a drawback that the deodorizing effect of the mate tea extract is significantly reduced under weakly acidic conditions. In order to enhance the deodorizing effect of mate tea extract in foods and drinks under mildly acidic conditions, it is necessary to use mate tea extract much more than the normal application amount, resulting in a deodorant composition Things are expensive. In addition, mate tea has a strong bitterness and astringency, and is easily subject to restrictions on the amount and flavor when commercialized. Therefore, it has been considered undesirable to use a large amount of mate tea extract in order to enhance the deodorizing effect from the viewpoint of practical use.

Japanese Patent Publication No. 2007-215831 Japanese Patent Publication No. 2007-000508 Japanese Patent Publication No. 8-127514 Japanese Patent Publication No. 7-124469 Japanese Patent No. 3341008 Patent Publication 2008-088102 Japanese Patent Publication No. 6-135848 Japanese Patent Publication No. 2009-190990

Negishi “Ingredients and functionality of bitter tea”: Journal of Japanese Society of Food Life, vol. 18, no. 1,25-31 Negishi et al., “Deodorization with Ku-ding-cha Containing a Large Amountof Caffeoyl Quinic Acid Dericatives”: J. Agric. Food Chem ,. Vol 52, No 17, 5513-5518 (2004) Miyamoto et al. “Deodorizing Action by Plant Extract”: Fragr. j. , No. 65, 82-85 (1984)

  The present invention relates to a deodorizing composition using a mate tea extract, and relates to a composition having a high deodorizing effect even under weak acidity.

As a result of intensive studies, the present inventors have found that a combination of mate tea extract and laccase exhibits a high deodorizing effect even under weakly acidic conditions.
That is, this invention relates to the deodorizing composition containing a mate tea extract and laccase.

  The deodorizing composition of the present invention has a high deodorizing effect even under conditions of weak acidity in which the deodorizing effect has been reduced in the conventional deodorant composition using mate tea extract.

It is the figure which compared the deodorizing effect of the mate tea water extract under weakly acidic conditions with the case where laccase is not added and the case where it is not added. It is a figure which shows each deodorizing effect of a mate tea and a strawberry tea water extract. It is a figure which shows the difference in the taste evaluation by the extraction conditions of a mate tea extract. It is a figure which shows each deodorizing effect of mate tea and chlorogenic acid.

The deodorant composition of the present invention uses mate tea extract as a main raw material.
Mate tea is a tea made from evergreen oak, Yerba mate leaves and twigs, which are widely used mainly in Brazil, Argentina, Paraguay and Uruguay, as coffee and tea. It is said to be a “drinking salad” because it contains a large amount of minerals and polyphenols, and is an important source of nutrient intake in some parts of South America where it is difficult to grow vegetables. They have abundant dietary experience, and record production has been conducted mainly in Paraguay since the end of the 15th century. Currently, the production volume is about 200,000 tons in Brazil, about 170,000 tons in Argentina, and about 40,000 in Paraguay. It reaches t.
As the mate tea extract, mate tea peel, leaves, pulp, fruits, materials, bark, roots, preferably dried leaves thereof are used. The method for obtaining the plant extract as an active ingredient of the present invention is not particularly limited, but the plant is pulverized by an appropriate pulverization means, and the extract is prepared by a method such as solvent extraction including two-stage extraction. As an extraction solvent, water or a lower alcohol such as methanol, ethanol, n-propanol and n-butanol, or one or more organic solvents such as ether, chloroform, ethyl acetate, acetone, glycerin and propylene glycol are mixed. Preferably, water or a hydrophilic organic solvent is used. In view of the fact that the extract of the present invention is often used as a food or drink, it is preferable to use a combination of water and ethanol as an extraction solvent from the viewpoint of safety. More preferably, the extraction is performed with 90% or less of ethanol, more preferably 60% or less of ethanol, further preferably 30% or less of ethanol, and most preferably with water.

Moreover, it is preferable to extract the mate tea extract in two stages because taste is improved.
Although the said extraction solvent can be used also as a solvent used for two-stage extraction, after extracting with 100% ethanol first, it is preferable to extract with water or 0-15% ethanol.

As extraction conditions, extraction can be performed at any one of high temperature, room temperature, and low temperature, but it is preferable to extract at 50 to 90 ° C. for about 1 to 5 hours. When the extraction is performed in two stages, it is preferable to first extract the first stage at 50 to 90 ° C. for about 1 to 5 hours, and then perform the second stage extraction at 50 to 90 ° C. for about 1 to 5 hours. The obtained extract may be filtered and the extraction solvent may be distilled off, followed by concentration or lyophilization under reduced pressure. In addition, those obtained by fractionating and purifying these extracts using an organic solvent, column chromatography, or the like can also be used.
In addition, since the deodorant composition of the present invention is highly safe, for example, a deodorant composition such as a mouthwash, a toothpaste, a deodorant spray, or a chewing gum, candy, tablet candy, gummy jelly, chocolate, biscuits. , Snacks and other confectionery, ice cream, sorbet, frozen confectionery such as ice confectionery, beverages, bread, hot cakes, dairy products, ham, sausage and other livestock products, sea bream, chikuwa and other fish products, side dishes, pudding, soup and It can be blended into foods and drinks such as jam and used on a daily basis. In addition to products put in the mouth, the deodorant composition of the present invention can be added to soaps, shampoos, rinses, creams, lotions, pet deodorants, indoor deodorants, indoor cleaning filters, toilet deodorants, etc. It is possible to make acidic products with excellent odor activity.
The blending amount may vary depending on various production conditions, but the mate tea extract is 0.01% by weight or more and 2.0% by weight or less, preferably 0.01% by weight or more and 1% with respect to the deodorant composition. 0.0 wt% or less, more preferably 0.05 wt% or more and 0.4 wt% or less, and the amount of laccase added varies depending on the titer of the laccase preparation, but is 0.01 wt% or more and 1.0 wt% or less, preferably It is preferable to blend 0.015 wt% or more and 0.12 wt% or less.
Moreover, when the addition amount of the said laccase is shown with a titer, laccase 5.90 * 10 < ² > U or more and 5.9 * 10 < ⁴ > U or less with respect to 100g of deodorant compositions, Preferably 3.0 * 10 < ³ > U or more 2 It is preferable to blend 4 × 10 ⁴ U or less.
Here, the titer of laccase is defined as 1 U unit of enzyme activity that increases the absorbance at 420 nm of the condensate produced by the oxidative condensation reaction of chlorogenic acid catalyzed by laccase at 0.001 per 30 seconds at 37 ° C. To do. 1U unit is obtained by adding 100 μl of 50 mM chlorogenic acid solution (0.1 M sodium phosphate buffer (pH 6.0)) to 500 μl of enzyme solution (0.1 M sodium phosphate buffer (pH 6.0)) The reaction was performed at 37 ° C. for 5 minutes. Thereafter, 500 μl of 0.1N sulfuric acid was added to stop the reaction, and the absorbance at 420 nm was measured. The enzyme activity that increases the absorbance at 420 nm 30 seconds after the start of the reaction by 0.001 was defined as 1 U unit.

The deodorizing composition according to the present invention exhibits a deodorizing effect even under weakly acidic conditions, in which the deodorizing effect is significantly reduced in the conventional deodorizing composition. That is, even if the pH in saliva after ingestion is in the range of 4.0 or more and 7.0 or less, the deodorizing composition of the present invention has a deodorizing effect.
Moreover, since this invention uses the mate tea conventionally utilized, even if it ingests in a body, there is no problem about the safety.

  The present invention will be further described below with reference to examples, but these examples do not limit the scope of the present invention.

Comparison of deodorizing effect when laccase was added to various tea leaf extracts The deodorizing effect when laccase was added to tea leaf extract was compared by the following procedure.

(Example 1-1) Reagent Methyl mercaptan sodium used the Tokyo Chemical Industry Co., Ltd. product. Methyl mercaptan standard solution (benzene solution) was manufactured by Wako Pure Chemical Industries, Ltd. As the laccase (EC 1.10.3.2), “Lacase Daiwa Y120” (trade name) manufactured by Daiwa Kasei Co., Ltd. was used.

(Example 1-2) Test plants The following dry plants were tested. Mate tea (Ilex paraguariensis) leaves, black tea leaves, oolong tea leaves and Rubus suavissimus leaves were obtained from Nippon Green Tea Center Co., Ltd. Green tea leaves were obtained from Nanzanen Co., Ltd.

(Example 1-3) Plant extraction method For each dry plant, 1 L of an extraction solvent (water or water-containing ethanol) was added to 50 g of powder, and reflux extraction was performed at 90 ° C for 1 hour. The extract was filtered, concentrated under reduced pressure, and lyophilized to obtain an extract.

(Example 1-4) Deodorant activity evaluation method for methyl mercaptan i) GC apparatus and analysis conditions The GC analysis conditions for preparing and quantifying a calibration curve for methyl mercaptan were the following apparatuses and conditions.
・ GC system: Agilent 7890A GC System (Agilent Technology Co., Ltd.)
Column: HP-5 (10 m × 0.53 mm, 2.65 μm, Agilent Technologies Inc.)
-Column flow rate: He, 10 ml / min
・ Split ratio: 15: 1
・ Inlet: Split, 200 ℃
・ Oven temperature: 50 ℃
・ Detector: FPD detector, 200 ℃
ii) Preparation of calibration curve A methyl mercaptan standard solution (benzene solution) was diluted with benzene, and a predetermined amount thereof was injected into the GC, and a calibration curve was created based on the peak height of the obtained methyl mercaptan.
iii) Evaluation method of deodorant activity After dissolving the sample in 900 μl of 0.2 M phosphate buffer (pH 4.5) in a 30 ml vial (control is buffer only), 0.2 M phosphate buffer is also used. Add 100 μl of 10 mg / ml laccase solution dissolved in (pH 4.5), add 500 μl of 25 ppm methyl mercaptan sodium aqueous solution, immediately plug with a Teflon (registered trademark) -coated rubber stopper, and stir. Shake for 5 minutes in a 37 ° C. water bath. After shaking, 50 μl of headspace gas was changed to GC, and the amount of methyl mercaptan (MS) when the sample was added and the amount of methyl mercaptan (MB) when no sample was added were determined, and the methyl mercaptan deodorization rate was calculated by the following equation. .
Methyl mercaptan deodorization rate (%) = (MB-MS) / MB × 100

(Example 1-5) Comparison of deodorizing effect Table 1 shows a comparison of the deodorizing effect on methyl mercaptan when laccase was not added to and added with 50 μg of various tea leaf extracts extracted under different extraction conditions. . Compared with other tea leaf extracts, mate tea extract had a very high deodorant effect, and the combination of mate tea water extract and laccase showed the strongest deodorant activity.

(Example 1-6) Deodorizing effect of pH and mate tea water extract About extract of mate tea water with methyl mercaptan deodorization rate at different pH by changing the pH of the reaction solution (Example 1-4) and It measured similarly.
The results are shown in FIG. Under mild acidity up to pH 4-7, the deodorization rate of the mate tea water extract was significantly increased by the addition of laccase.

Comparison of deodorizing effect between mate tea and strawberry tea In order to compare the deodorizing effect of a known deodorant composition (Patent Document 8) containing a strawberry case extract of rosaceae and laccase, (Examples) The following experiment was conducted using the extract of mate tea water extracted in 1-3) and the extract of strawberry tea water.

(Example 2-1) Measurement of 50% effective concentration (EC ₅₀ ) With respect to the extract of mate tea and strawberry tea water, the deodorization rate at each concentration was obtained in the same manner as in Example 1 except that the sample concentration was changed. It was. Thereafter, a graph with the logarithm of the sample concentration as the horizontal axis and the deodorization rate as the vertical axis was prepared, and the concentration (EC ₅₀ ) at which 50% deodorization rate was obtained was obtained.
The prepared graph is shown in FIG. 2 and Table 2 shows the deodorizing effect of mate tea and tea tea water extract.
2. From FIG. 2, it was recognized that mate tea has a deodorizing effect on methyl mercaptan at a lower concentration than strawberry tea. Further, from Table 2, EC ₅₀ of mate tea was 20.53 μg / ml, and a deodorizing effect three times or more than that of strawberry tea was recognized.

Mate Tea Extraction Condition and Deodorizing Effect In order to evaluate the difference in odor eliminating effect depending on the mate tea extraction condition, methylate was extracted in the same manner as in Example 1 except that mate tea was extracted under different extraction conditions as follows. The deodorizing activity for mercaptans was evaluated.

(Example 3-1) Plant extraction method i) One-step extraction 1 L of extraction solvent was added to 50 g of dried plant powder, and reflux extraction was performed at 90 ° C for 1 hour. The extract was filtered, concentrated under reduced pressure, and lyophilized to obtain an extract. As the extraction solvent, water or water-containing ethanol with different concentrations was used.
ii) Two-stage extraction 1 L of ethanol was added to 50 g of dry powder of mate tea, reflux extraction was performed at 90 ° C. for 1 hour, and the extract was separated by filtration. Water or 1 L of 15% aqueous ethanol was added to the extraction residue obtained by filtration, and reflux extraction was performed at 90 ° C. for 1 hour. The extract was filtered, concentrated under reduced pressure, and lyophilized to obtain an extract.

(Example 3-2) Comparison of deodorizing effect Table 3 shows the deodorizing effect on methyl mercaptan when laccase was added to 20 µg of mate tea extract with different extraction conditions. In one-stage extraction, mate tea 15% ethanol extract showed the strongest deodorizing activity, followed by water extract, and the tendency was that deodorizing activity tended to be higher when the extraction solvent was aqueous. . In addition, in the case of strawberry tea belonging to the family Rosaceae, it is reported that the deodorizing effect is improved by performing two-stage extraction with water extraction after pretreatment with 100% ethanol (Patent Document 8). Therefore, mate tea was also subjected to two-stage extraction in which pretreatment with 100% ethanol was performed to evaluate the deodorizing effect. As a result of the test, the two-stage extraction did not significantly improve the deodorizing effect, but no decrease was observed.

Difference in taste depending on mate tea extraction conditions In order to evaluate the difference in taste due to mate tea extraction conditions, the following sensory test was performed using the mate tea extract extracted in Example 3 above with laccase added. went.

(Example 4-1) Sensory test A 0.25% (wt / v) aqueous solution was used for the sensory test of the mate tea extract. The inspection items were “bitterness”, “astringency”, “aftertaste”, “acidity”, and “sweetness”, and taste evaluation was performed according to the evaluation criteria shown in Table 4 by five panelists.

(Example 4-2) Comparison of taste The taste evaluation of the mate tea extract (0.25% aqueous solution) by five panelists is shown in FIG. The higher the ethanol concentration of the extraction solvent, the higher the “bitter taste” and “astringency” scores. It was also confirmed that when the two-step extraction process was performed on the water extract and the 15% ethanol extract, the scores of “bitter taste” and “astringency” decreased.

Ingredient composition and deodorizing effect of mate tea extract and influence on taste In order to investigate the influence on taste of ingredient composition in mate tea extract, the following analysis was performed using the mate tea extracted in Example 3 above. The contents of chlorogenic acid, caffeine and caffeic acid contained in mate tea extracted under each extraction condition were analyzed.

(Example 5-1) Reagent Methyl mercaptan sodium and chlorogenic acid were manufactured by Tokyo Chemical Industry Co., Ltd. Methyl mercaptan standard solution (benzene solution), caffeine, caffeic acid and ursolic acid were manufactured by Wako Pure Chemical Industries, Ltd. Laccase (EC 1.10.3.2) trade name “Lacase Daiwa Y120” manufactured by Daiwa Kasei Co., Ltd. was used. Acetonitrile, methanol, phosphoric acid, acetic acid, distilled water were used for HPLC, and other reagents used were commercial grades.

(Example 5-2) Quantification of chlorogenic acid, caffeine, and caffeic acid i) HPLC apparatus and analytical conditions HPLC analysis for preparing and quantifying calibration curves for chlorogenic acid, caffeine, and caffeic acid was performed using the following apparatus. Performed under conditions.
・ HPLC equipment: Alliance HPLC (Waters)
・ Column: Cadenza CD-C18 (4.6 × 250 mm, 3 μm, intact)
Guard column: Cadenza CD-C18 (2 × 5 mm, 3 μm, intact)
-Mobile phase: A 0.5% (v / v) acetic acid aqueous solution, B methanol, C acetonitrile
0-15min A: B: C = 70: 30: 0
15-30min A: B: C = 0: 0: 100
・ Detector: UV detector (274 nm)
・ Temperature: 35 ℃
・ Flow rate: 0.7 ml / min
・ Injection volume: 2 μl
ii) Preparation of calibration curve A solution obtained by dissolving chlorogenic acid, caffeine, and caffeic acid in 0.5% (v / v) acetic acid aqueous solution and appropriately diluting with 0.5% (v / v) acetic acid aqueous solution is used as a membrane filter. (0.45 μm), a fixed amount of each standard solution was injected into the HPLC, and a calibration curve was prepared based on the obtained peak area.
iii) Preparation of test solution 5 ml of methanol was added to 10 mg of mate tea extract and dissolved, and the volume was adjusted to 10 ml with 0.5% (v / v) acetic acid aqueous solution, followed by filtration with a membrane filter (0.45 μm). The filtrate was used as a test solution.
iv) Quantification HPLC analysis was performed with the above apparatus using the obtained test solution, and the respective contents were quantified based on the calibration curve.

(Example 5-3) Quantification of saponin The saponin of the mate tea extract was determined by measuring the amount of ursolic acid after acid hydrolysis and converting it to matesaponin 1.
i) HPLC apparatus and analysis conditions HPLC analysis for the preparation and quantification of ursolic acid calibration curve was performed with the following apparatus and conditions.
・ HPLC equipment: Alliance HPLC (Waters)
・ Column: Cadenza CD-C18 (4.6 × 250 mm, 3 μm, intact)
Guard column: Cadenza CD-C18 (2 × 5 mm, 3 μm, intact)
-Mobile phase: A distilled water, B acetonitrile
A: B = 10: 90
・ Detector: UV detector (203 nm)
・ Temperature: 25 ℃
・ Flow rate: 1.0 ml / min
・ Injection volume: 2 μl
ii) Preparation of calibration curve Dissolve ursolic acid in acetonitrile, filter the solution diluted with acetonitrile as appropriate with a membrane filter (0.45 μm), inject a fixed amount of each standard solution into HPLC, and determine the peak area obtained. A calibration curve was created.
iii) Preparation and quantification of test solution Dissolve mate tea extract in water to prepare a 5 mg / ml aqueous solution, take 10 ml of this solution in a screw-cap test tube, add 1.5 ml of 4M HCl, and vortex well. Thereafter, the mixture was refluxed at 110 ° C. for 2 hours. After refluxing, the mixture was cooled at room temperature, extracted with chloroform (5 ml × 4 times), and chloroform was completely removed by drying under reduced pressure. After making it dry, it melt | dissolves in acetonitrile, After making constant volume to 10 ml, it filtered with the membrane filter (0.45 micrometer), and used the filtrate as the test solution.
Using the obtained test solution, HPLC analysis was performed with the above apparatus, and the amount of saponin was determined by converting into matesaponin 1 according to the following formula based on the amount of ursolic acid (C _u ) in the obtained sample.
Saponin content (% (wt / wt)) = C _u × (MW _s / MW _u )
MW _s : molecular weight of matesaponin 1 (MW = 913.108)
MW _u : molecular weight of ursolic acid (MW = 456.711)

(Example 5-4) Ingredient composition Table 5 shows the content of chlorogenic acid as a polyphenol component, caffeine, caffeic acid, and saponin as taste components in a mate tea extract. Since the caffeine content and the saponin content are low in the two-stage extract, it is considered that improvement was recognized in the items of “bitter taste” and “astringency” in the taste evaluation.

Contribution of chlorogenic acid in the deodorizing effect of mate tea extract In order to determine the chlorogenic acid contribution in 15% ethanol extract, which was the most active in mate tea extract in Example 3, mate tea was used in the same manner as in Example 2. The 50% effective concentration (EC ₅₀ ) of the extract and chlorogenic acid was measured, and the contribution rate of chlorogenic acid to the deodorizing activity was determined. A graph with the logarithm of the sample concentration on the horizontal axis and the deodorization rate on the vertical axis is shown in FIG. 4, and the 50% effective concentration (EC ₅₀ ) of mate tea extract and chlorogenic acid and the contribution ratio of chlorogenic acid are shown in Table 6. The contribution rate was calculated from each EC ₅₀ by the following formula.
Contribution rate = _{(EC 50} of _{EC 50} / chlorogenic acids mate) × (chlorogenic acid content in the mate (%) / 100)
From Table 6, since the contribution rate of chlorogenic acid is as low as 12.3%, it is considered that polyphenol components other than chlorogenic acid are also involved in the activity.

Trial Production of Mate Tea Extract, Citric Acid, and Laccase Chewing Gum The raw materials described in the following formulation examples containing laccase and citric acid were mixed with the prepared strawberry tea extract, and chewing gum was obtained by a conventional method.
Moreover, as a control example, the raw materials described in the following prescription examples containing only the prepared strawberry tea extract were mixed, and chewing gum was obtained by a conventional method.

Produced gum garlic odor control effect (human test)
(Example 8-1) Sensory evaluation method The sensory evaluation value was evaluated according to Table 8 below, and the values of five panelists were averaged and digitized. After collecting the bad breath of the subject 10 minutes before the test, 6 pieces of dumplings were ingested for 2 minutes, and the gum in Example 7 and the control example (chewing for 5 minutes) or gargle (washed with 20 ml of water twice). The breath immediately after ingestion was collected in a smelly bag, and the degree of bad breath was evaluated. The results are shown in Table 9. It was confirmed that the gum of Example 7 suppressed the garlic odor most strongly after ingestion. The saliva pH after ingesting the gum of Example 7 was about 6.0.

  Using the extract prepared by the method shown in Example 1-3, candy, gummy jelly, and troche were produced according to the following formulation.

(Example 9-1)
Candy prescription sugar 50.0% by weight
Mizuame 34.0
Citric acid 2.0
Mate tea extract 0.2
Laccase formulation 0.1
Fragrance 0.2
Water remaining
100.0

(Example 9-2)
Gummy jelly prescription gelatin 60.0% by weight
Mizuame 21.0
Sugar 8.5
Vegetable oil 4.5
Mannitol 3.0
Malic acid 2.0
Mate tea extract 0.2
Laccase formulation 0.2
Fragrance 0.6
100.0

(Example 9-3)
Troche prescription glucose 72.3% by weight
Lactose 15.0
Gum arabic 6.0
Fragrance 1.0
Sodium monofluorophosphate 0.7
Mate tea extract 2.0
Laccase formulation 1.0
Lactic acid 2.0
100.0

Claims (3)

  It is characterized by adding 100% ethanol to the peel, leaf, pulp, fruit, wood, bark, root or leaf of mate tea and extracting it with water or 0-15% aqueous ethanol. The manufacturing method of the deodorizing composition containing a mate tea extract and laccase. The content of the mate tea extract in the deodorant composition is 0.01% by weight or more and 2.0% by weight or less , and the content of the laccase in the deodorant composition is 0.01% by weight or more and 1.0% by weight. %. The method for producing a deodorant composition according to claim 1, wherein the deodorant composition is further prepared so as to be not more than%. A method for producing a chewing gum, candy, tablet, or gummy jelly, comprising the deodorant composition obtained by the production method according to claim 1 or 2 .

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