JP2006345724A - Peroxynitrite-eliminating agent, and food and drink, healthy tea leaf and healthy drink respectively containing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a peroxynitrite-eliminating agent exerting excellent peroxynitrite-eliminating function, food and drink containing the agent, and healthy tea leaves and healthy drink exerting excellent peroxynitrite-eliminating function and containing no caffeine. <P>SOLUTION: The peroxynitrite-eliminating agent contains okra or okra seeds as active ingredients. The food and drink contain the peroxynitrite-eliminating agent. The healthy tea leaves contain roasted okra or okra seeds as main raw material. The healthy tea leaves comprise a combination of the roasted okra or okra seeds and at least one kind of aloe, turmeric, barley, persimmon leaves, striped bamboo, black beans, mulberry leaves, brown rice, adlay, Cassia tora, barley tea, bracket fungus of the genus Fomes, dietary fiber and orange peel. The healthy drink is produced by diluting or leaching out the healthy tea leaves. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention not only prevents the deterioration of various symptoms by suppressing the production of peroxynitrite, which causes various diseases such as lifestyle-related diseases and aging in the body, but also prevents aging in the body. The present invention relates to a peroxynitrite scavenger useful for maintaining health and foods and drinks containing the same. The present invention also relates to a healthy tea leaf that does not contain caffeine as a component and exhibits a high peroxynitrite erasing action, and a health drink produced thereby.

  In recent years, reactive oxygen species such as superoxide, hydrogen peroxide, singlet oxygen, hydroxy radical, nitric oxide, and peroxynitrite oxidize DNA, proteins, lipids, etc. in the body, various cancers, heart diseases, strokes, It has been clarified that it causes various diseases such as diabetes and other lifestyle-related diseases.

Among these active oxygen species, peroxynitrite has a very strong oxidizing power and has a long half-life even under physiological conditions. Therefore, it is said that pathological factors increase when it exists in vivo. Peroxynitrite causes many biological disorders such as protein, aromatic amino acid nitration, hydroxylation, DNA cleavage, and lipid peroxidation. In addition, it has been reported that ONOOCO ₂ ⁻ is generated by reaction with carbon dioxide (CO ₂ ), which is abundant in living organisms, and exhibits higher reactivity. Therefore, a substance having a strong peroxynitrite scavenging activity has attracted much attention in terms of preventing the pathological condition caused by peroxynitrite and maintaining its health, and is considered to be promising as a new health food material.

  Green tea is an example of a food having a high peroxynitrite-erasing action. However, green tea contains a large amount of caffeine, so caffeine intake cannot be avoided. Caffeine has an action to excite the central nervous system and is said to be effective in preventing drowsiness, fatigue and headache, and is also used as a medicine. However, due to the physiological effects of caffeine, symptoms such as arrhythmia, dizziness, and insomnia may occur depending on the amount consumed and individual differences, especially in pregnant women, since caffeine passes through the placenta and should be avoided. It is also recommended to avoid children and people with impaired liver function. For these reasons, it is desired to have a low caffeine content and a strong peroxynitrite erasing action.

  In addition, for okra, as a problem in production areas, things that are damaged by typhoons, etc., those that are deformed, those that are out of specification, etc. cannot be distributed as products. It is rare.

  As for okra, it has come to be used in cosmetics such as hyaluronic acid synthesis accelerators and whitening cosmetics as one of cosmetic ingredients (see, for example, Patent Documents 1 and 2). As for okra seeds, there is an example in which a protein fraction extracted from okra seeds is used as a cosmetic (see, for example, Patent Document 3). However, these are mainly examples of cosmetic ingredients, and no peroxynitrite erasing effect of okra and okra seeds and no patents relating to foods and drinks using the effect have been reported.

  On the other hand, regarding the production of beverages of okra, a patent relating to a production method for reducing the slime when a juice extract of vegetables having a slime such as okra is prepared has been reported (for example, see Patent Document 4). Moreover, as a manufacturing method of green vegetable juice, what described okra as one of the materials of green vegetables is mentioned (for example, refer patent document 5). However, there is no example of applying okra or okra seeds to drinks such as tea.

JP 2004-51533 A JP 2003-246722 A JP-T-2001-518910 JP 2000-228962 A JP-A-8-308542

In view of the above points, the present invention effectively uses okra that is not distributed as a product for reasons such as non-standards in production areas and the like, and a peroxynitrite erasing agent that can exhibit a high peroxynitrite erasing action and its The purpose is to provide food and drinks.
Another object of the present invention is to provide a healthy tea leaf that does not contain caffeine, which is a problem with green tea, and that can exhibit a high peroxynitrite-erasing action and its beverage.

In order to achieve the above object, the present inventors have conducted extensive research and found that okra (the whole edible portion of okra) or okra seeds have peroxynitrite scavenging activity. Was found to have stronger peroxynitrite scavenging activity, and the present invention was completed.
That is, according to the present invention, a peroxynitrite scavenger containing okra or okra seeds as an active ingredient is provided, and a food or drink containing the peroxynitrite scavenger is provided.
In addition, health tea leaves mainly made from roasted okra or okra seeds are provided, and health drinks produced by decocting or leaching the health tea leaves are also provided.

The peroxynitrite scavenger of the present invention contains okra or okra seeds as an active ingredient and has excellent peroxynitrite scavenging activity.
Moreover, since the food / beverage products using the okra or okra seeds of the present invention include okra or okra seeds exhibiting excellent peroxynitrite erasing activity, an excellent peroxynitrite erasing effect can be expected.
Moreover, healthy tea leaves and health drinks produced from okra or okra seeds do not contain caffeine and have high peroxynitrite scavenging activity.
Furthermore, when implementing the present invention, it is possible to effectively utilize okra which has been conventionally regarded as waste.

  For the preparation of the peroxynitrite scavenger in the present invention, a normal commercially available okra can be used, but an okra that is disposed of as non-standard in the production area or the like can also be used.

  Examples of the peroxynitrite scavenger in the present invention include the following.

  That is, okra or okra seeds may be either themselves or a pulverized product (raw or dried product) thereof, or may be extracted with hot water or an organic solvent. Examples of the organic solvent used for extraction include methanol, ethanol, acetone, ethyl acetate or their hydrates, chloroform, dichloromethane, pentane, hexane, heptane and the like. Among these, methanol, ethanol, hexane and the like are preferable. .

  The resulting extract may be concentrated and the concentrate may be dried. Further, the obtained extract may be purified before and after concentration, and a chromatographic method or the like can be used as the purification operation.

  The okra or okra seed peroxynitrite-eliminating agent can be made into various forms such as granules, tablets, capsules and the like by known methods as food and drink products.

  Okra or okra seeds can be used as a food and drink for the purpose of maintaining and promoting health and wellness by using the high peroxynitrite-erasing action of okra or okra seeds. Liquid, semisolid or solid products such as soft drinks and soup Etc.

  In addition, okra or okra seeds may be used as okra or okra seed tea, because the peroxynitrite erasing ability is not significantly impaired by roasting or the like.

  For the preparation of healthy tea leaves in the present invention, normal commercially available okra can be used, but okra that is disposed of as non-standard in the production area or the like can also be used. The okra or okra seed may be either itself or a pulverized product thereof (raw or dried product), but when making healthy tea leaves using okra, it is preferably not pulverized product.

  When making healthy tea leaves using okra itself, if it is finely pulverized, it will become sticky during roasting, so it is preferable to dry the whole without cutting. Also, there is no need for boiling. Moreover, in order to eliminate the bacteria which exist, it is preferable to heat at 130 degreeC or more.

  The health tea leaves or the leachate thereof according to the present invention can be used not only as a single use thereof, but also as a tea leaf or a health drink as a mixture with other teas or the leachate thereof. In this case, other teas or their exudates include aloe, turmeric, barley, birch leaves, bears, black beans, mulberry leaves, brown rice, hato barley, hub tea, barley tea, ganoderma, dietary fiber, mandarin orange Examples include leather.

  Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples.

Example 1: Okra or okra seed powder and extract Okra or okra seed powder, hot water extract and ethanol extract were prepared using the following methods.

1) Okra powder 12.5 kg of okra powder was frozen, water was evaporated with a freeze dryer, freeze dried, and then pulverized with a pulverizer to obtain 1.25 kg of okra powder.
2) Okra seed powder 600 g of okra seed was frozen, water was evaporated with a freeze dryer, freeze dried, and then pulverized with a pulverizer to obtain 78 g of okra seed powder.
3) Okra hot water extract 3 g of the above okra powder was soaked in 90 mL of hot water and extracted with stirring for 15 minutes. The residue was filtered off, the filtrate was frozen, and the water was evaporated in a freeze dryer to obtain 854 mg of okra hot water extract.
4) Okra seed hot water extract 3 g of the above okra seed powder was dipped in 90 mL of hot water and extracted by stirring for 15 minutes. The residue was filtered off, the filtrate was frozen, and water was evaporated in a freeze dryer to obtain 1469 mg of okra seed hot water extract.
5) Okra ethanol extract 1 g of the above okra powder was dipped in 200 mL of ethanol and extracted with stirring overnight. The residue was filtered off and dried under reduced pressure on a rotary evaporator to obtain 222 mg of okra ethanol extract.
6) Okra seed ethanol extract 1 g of the above okra seed powder was dipped in 200 mL of ethanol and extracted with stirring overnight. The residue was filtered off and dried under reduced pressure on a rotary evaporator to obtain 155 mg of okra seed ethanol extract.

Comparative test 1: Peroxynitrite erasing activity measurement 1) Sample preparation Okra powder, okra seed powder, okra hot water extract, okra seed hot water extract, okra ethanol extract and okra seed ethanol extract prepared by the above method In addition, using BHT and α-tocopherol as samples, 10 mL of methanol was added to 10 mg of each sample, and this was further diluted 2-fold with methanol to obtain a sample solution. Methanol was used as a control.

2) Peroxynitrite scavenging activity measurement method Tyrosine is nitrated by peroxynitrite to form nitrotyrosine. The area of the nitrotyrosine peak when the control solution was used was defined as an erasure rate of 0, and the state where no peak was detected was defined as an erasure rate of 100. Higher erasure rate means higher peroxynitrite erasing power.

(1) Reagents a) Synthesis of peroxynitrite: Put 200 mL of ice-cooled 1.5M NaOH into an absorption bottle and suck it, and suck it in advance and add 0.6M HCl 100mL containing 0.6MH ₂ O ₂ and 0.6M NaNO ₂ 100mL. Was poured at the same time. The reaction solution was put into a freezer, and when used, the upper concentrated solution generated at the stage where the reaction solution was removed from the freezer and thawed a little was used. H ₂ O ₂ present in the concentrate was removed with manganese dioxide and used.
b) As for the L-tyrosine solution, 72 mg of L-tyrosine and 3.36 mg of Fe (III) -EDTA were added to 200 mL of 100 mM phosphate buffer (pH 6.4) and dissolved with an ultrasonic cleaner.

(2) Measuring method
1 mL of the sample solution was added to 1 mL of the L-tyrosine solution, and 20 μL of peroxynitrite was added, followed by stirring for 5 minutes. The area of nitrotyrosine produced in the reaction solution was measured using HPLC.

(3) Analysis conditions
The analysis conditions for HPLC analysis were as follows (repeated three times).
Column: Capsule PAK, C18 AG120 (4.6 × 250)
Guard column: ODS-120A
Eluent: 50 mM pH 6.4 Phosphate buffer: Methanol = 97: 3
Flow rate: 0.9mL / min
Detector: UV absorptiometer (measurement wavelength 280nm)
Column temperature: 40 ° C
Injection volume: 20μL

(4) Measurement results The measurement results were as follows.

  As shown in Table 1 above, okra powder, okra seed powder, okra hot water extract, okra seed hot water extract, okra ethanol extract and okra seed ethanol extract have strong peroxynitrite scavenging activity. It was also confirmed that the okra seed powder and extract had a stronger scavenging activity than α-tocopherol and comparable to BHT.

Example 2: Food / beverage products Tablets of okra or okra seeds were prepared using the following method.
1) Okra tablets 50 parts by weight of okra powder, 200 parts by weight of sucrose, and 10 parts by weight of glycerin fatty acid ester were mixed and mixed and tableted by methods known in the art to produce tablet-shaped foods.
2) Tablets of okra seeds Okra seed powder 50 parts by weight, sucrose 200 parts by weight, and glycerin fatty acid ester 10 parts by weight were mixed and mixed and tableted by a method known in the art to produce tablet-shaped foods.

Example 3 Food / Beverage Products Okra or okra seed soup was prepared using the following method.
1) Okra soup A soup was prepared at a ratio of 6% okra powder, 20% chicken, 5% vegetables, 6% seasonings and 63% water.
2) Okra Seed Soup A soup was prepared in a ratio of 0.5% okra seed powder, 25% pumpkin, 7% onion paste, 29% processed milk, 7% seasonings and 31.5% water.

Example 4: Food and beverage products Okra or okra seed drinks were prepared using the following method.
1) Okra beverage (sport drink type)
Okra powder 0.5g, liquid sugar 1g, xylitol 1g, sodium chloride 0.05g, potassium chloride 0.03g, monosodium phosphate 0.02g, anhydrous citric acid 0.65g, aspartame 0.05g, flavor 0.23g, water 96.47g A beverage was prepared.
2) Okra seed beverage (sports drink type)
Okra seed powder 0.5g, liquid sugar 1g, xylitol 1g, sodium chloride 0.05g, potassium chloride 0.03g, monosodium phosphate 0.02g, anhydrous citric acid 0.65g, aspartame 0.05g, flavor 0.23g, water 96.47g An okra seed drink was prepared.

Example 5: Healthy tea leaves and healthy beverages 27 kg of raw okra were heat-dried with a drum heater (300 ° C., 1 hour 20 minutes, product temperature 95 ° C.). Then it was cooled and left for 1 hour. Thereafter, drying under reduced pressure was carried out at 60 ° C. for 3 hours. It was dried with a drum heater (300 ° C., 20 minutes, product temperature 135 ° C.). Thereafter, the mixture was pulverized by a pulverizer to obtain 2.6 kg of okra tea. When 10 g of this okra tea was eluted with 500 mL of hot water for 2 to 3 minutes, a good okra tea sufficiently containing the active ingredients was obtained (see Table 2 below).

Example 6: Healthy tea leaves and health drink 200 g of raw okra seeds were heated and roasted for 15 minutes with a roaster. Thereafter, the mixture was pulverized by a pulverizer to obtain 27 g of okra seed tea. When 10 g of this okra seed tea was eluted with 500 mL of hot water for 2 to 3 minutes, a good okra tea containing sufficient active ingredients was obtained (see Table 2 below).

Example 7: Healthy tea leaves and health drinks Roasted okra tea of Example 5 and other teas were mixed in the proportions shown in Table 2 to prepare healthy tea. When 10 g of this healthy tea was eluted with 500 mL of hot water for 2 to 3 minutes, a good okra tea sufficiently containing active ingredients was obtained (see Table 2).

Example 8:
The roasted okra seed tea of Example 6 and other teas were mixed in the proportions shown in Table 2 to prepare healthy tea. When 10 g of this healthy tea was eluted with 500 mL of hot water for 2 to 3 minutes, a good okra tea sufficiently containing active ingredients was obtained (see Table 2).

Comparative test 2: Peroxynitrite erasing activity and caffeine content measurement The peroxynitrite erasing activity of Examples 5 to 8 and Comparative Example 1 (green tea) was measured. The sample solution was prepared in the following manner, and the measurement method was in accordance with Comparative Test 1 above.
1) Sample preparation 100 mL of hot water was added to 2 g of each sample and filtered after 2 minutes. The filtrate was diluted 40 times with methanol and measured. Methanol was used as a control.

Moreover, the amount of caffeine contained in Examples 5 to 8 and Comparative Example 1 (green tea) was measured.
1) Sample preparation 100 mL of hot water was added to 2 g of each sample and filtered after 2 minutes, and this filtrate was used for measurement. Caffeine (10 mg / 100 mL) was used as the standard solution, and the concentration (mg / 100 mL) of the sample solution was measured. Caffeine was analyzed by HPLC.
2) Analysis conditions
The HPLC analysis conditions were as follows (repeated three times).
Column: Capsule PAK, C18 UG120 (4.6mm x 100mm)
Guard column: ODS-120A
Eluent: 0.5v / v% phosphoric acid: methanol = 84: 16
Flow rate: 0.8mL / min
Detector: UV absorptiometer (measurement wavelength 280nm)
Column temperature: 40 ° C
Injection volume: 10 μL

Table 3 shows the results of measuring the peroxynitrite erasing activity and the amount of caffeine contained.

  As shown in Table 3, tea using okra or okra seeds showed high peroxynitrite scavenging activity, and Examples 6 and 8 showed high scavenging activity almost comparable to green tea. Moreover, it was confirmed that tea using okra or okra seeds does not contain caffeine.

Claims (9)

A peroxynitrite scavenger comprising okra or okra seeds as an active ingredient. The peroxynitrite scavenger according to claim 1,
A peroxynitrite scavenger characterized by using okra or crushed okra seeds as a main raw material. The peroxynitrite scavenger according to claim 1,
A peroxynitrite erasing agent characterized in that the main raw material is okra or an okra seed pulverized product extracted with hot water or an organic solvent. The peroxynitrite scavenger according to claim 1,
A peroxynitrite scavenger characterized by using as a main raw material an extract obtained by extracting okra or a ground product of okra seeds with hot water or an organic solvent. The peroxynitrite scavenger according to claim 1,
A peroxynitrite scavenger characterized in that a main raw material is an extract obtained by extracting an okra or a ground product of okra seeds with hot water or an organic solvent and concentrating to dryness. A food or drink comprising the peroxynitrite scavenger according to any one of claims 1 to 5. Healthy tea leaves characterized by using okra or roasted okra seeds as a main ingredient. Okra or roasted okra seeds and aloe, turmeric, barley, birch leaves, bear cocoons, black beans, mulberry leaves, brown rice, hato barley, hub tea, barley tea, ganoderma, dietary fiber and orange peel A healthy tea leaf comprising a combination of at least one of them. A health drink produced by brewing or leaching the health tea leaves according to claim 7 or 8.