JPH07268322A - Antioxidation agent - Google Patents

Abstract

PURPOSE:To obtain an antioxidation agent containing a compound extracted from tamarind testa and having a specific structure as an active component, exhibiting antioxidation activity and useful for preventing the oxidative deteriora tion of the quality of foods, feeds, cosmetics, pharmaceuticals, etc. CONSTITUTION:This antioxidation agent contains a compound of formula I (R is methoxycarbonyl, acetyl or hydroxymethylcarbonyl) as an active component. The compound can be produced by extracting the testa of Tamarindus indica L. (an evergreen tree native to tropical district and belonging to the family Legminosae) preferably with ethyl acetate. Examples of the compound of formula I are compounds of formula II and formula III. The amount of the compound in the agent is 0.03-1wt.%, preferably 0.05-0.5wt.%.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antioxidant derived from a natural product, which has an antioxidant activity useful for preventing deterioration of quality of foods, feeds, cosmetics, pharmaceuticals or raw materials thereof due to oxidation.

[0002]

2. Description of the Related Art Conventionally, as an antioxidant, for example, BH
A (butylhydroxyanisole), BHT (butylhydroxytoluene), sodium erythorbate, isopropyl citrate, dl-α-tocopherol, propyl gallate and the like are used. However, in recent years
Due to increasing consumer awareness of the safety of food additives consisting of chemically synthesized products, the use of chemically synthesized products such as BHA and BHT tends to be avoided, and other than those that are considered to be relatively safe. It is difficult to find any effective antioxidant. Because of this,
Many attempts have been made to obtain highly safe and sufficiently effective antioxidants from natural products, and some have been put to practical use. For example, as antioxidants derived from natural products currently used in food processing, tocopherols contained in various plant oils, flavones contained in various plant tissues, quercetin, flavone derivatives such as rutin, tea leaves, and coffee. Beans, caffeic acid contained in cacao beans, gallic acid, gallic acid derivatives such as ferulic acid and polyphenols of catechins, gossypol contained in cottonseed oil, γ-oryzanol in rice bran, sezamol / sezamoline in sesame oil, various spices And the like, and protein hydrolysates (peptides, amino acids) and the like (for example, JP-A-59-453).
85, JP-A-59-166585, JP-A-60-3.
No. 6584, Japanese Patent Publication No. 61-34792, JP-A No. 3-7.
565, etc.).

[0003]

However, the above-mentioned antioxidants derived from natural products are not yet sufficiently satisfactory.
Furthermore, the development of new antioxidants that exhibit excellent effects has been desired. While conducting various searches and studies on antioxidant components in natural products, the present inventors surprisingly have excellent antioxidant activity from tamarind seed coat which has not been used as a source of antioxidants until now. The compound was successfully isolated and identified, and the present invention was completed.

[0004]

That is, the present invention provides the formula (1):

[0005]

[Chemical 4]

A compound represented by the formula (wherein R represents a methoxycarbonyl group, an acetyl group or a hydroxymethylcarbonyl group), particularly an antioxidant comprising a compound of the formula (1) derived from tamarind seed coat as an active ingredient. It is provided. In the compound of formula (1), R is methoxycarbonyl, that is, formula (2):

[0007]

[Chemical 5]

The compounds other than the compound represented by the formula are novel compounds which have not been described in the literature, and the present invention also provides a compound of the formula (3):

[0009]

[Chemical 6]

Further, equation (4):

[0011]

[Chemical 7]

There is also provided a compound of formula Hereinafter, the present invention will be described in detail. The compounds of formula used as an active ingredient (1) of the antioxidant of the present invention, extracted from seed coat of tropical native leguminous evergreen tree tamarind (Tamarindus indica L.), obtained by isolation and purification. Tamarind is available as a general food in the tropics, and its seed mucilage is used for adjusting the viscosity of foods and the like. The compound of formula (1) is obtained from tamarind seed coat as follows, for example. First, tamarind seed coat is pulverized by a known method, this is extracted with an appropriate amount of a solvent, and the extract is concentrated to obtain a crude extract. A polar or non-polar solvent such as ethyl acetate or ethanol can be used as the extraction solvent, and ethyl acetate is preferred. The extraction conditions are not particularly limited, but may be 8 to 1 at normal temperature to 30 ° C. under normal pressure to reduced pressure.
Extraction can be performed efficiently by performing extraction for 6 hours 2 to 4 times.
A known method is used for concentration, but vacuum concentration at room temperature to 40 ° C. is preferable from the viewpoint of less damage to the extract.

The crude extract obtained is further subjected to purification by high performance liquid chromatography. The conditions of high performance liquid chromatography are not particularly limited as long as the compound of the present invention can be isolated and purified. For example, a silica gel-based one as a separation column and a trifluoroacetic acid-containing acetonitrile / water as a solvent are used. Appropriate. The compound is preferably detected by ultraviolet absorption at 280 nm because the compound of formula (1) has a benzene ring. From the fraction containing the compound of formula (1), the solvent is removed by, for example, drying under reduced pressure to obtain the isolated and purified compound of formula (1).

The compound of formula (1) thus obtained may be used as an antioxidant as it is, or may be used in the form of a crude ethyl acetate extract from tamarind seed coat.
Alternatively, with these, known excipients acceptable for foods, cosmetics, pharmaceuticals and the like, and other appropriate additives, for example, lactose, dextrin, gums, glyceric acid ester and the like, by a method known per se. It can also be used in solid or liquid form, for example in the form of powders, pastes, emulsions, oil solutions, aqueous solutions. The amount of the compound of the formula (1) in such a formulation is appropriately selected depending on the intended use, the intended antioxidant action, etc., but is usually 0.03 to
Up to 1% by weight, preferably 0.05 to 0.5% by weight.

The compound of formula (1) is obtained from the seed coat of a natural product used for food and is considered to have extremely low toxicity. Therefore, the antioxidant of the present invention is used in foods, feeds, cosmetics and pharmaceuticals. Also, it can be used as an antioxidant for those raw materials and the like. For example, added in the manufacturing process of food etc.,
It can be used by mixing, wetting or other methods. The amount used can be appropriately selected depending on the desired purpose, but usually the concentration of the compound of formula (1) is adjusted to 0.05 to 0.5% by weight in the desired product.

The present invention will be specifically described below with reference to examples, but the present invention is not limited thereto. The antioxidant activity was measured using a model system using linoleic acid as a substrate [T. Osawa and M. Namiki (M. N.
amiki), Agricultural and Biological Chemistry (Agric. Biol. Chem.), Vol. 45, 73.
5 (1981)], using the Rodan iron method [Mitsuda et al., Nutrition and Food Vol. 19, 210 (1966)] and TB.
Method A [O. Ottolenghi, Achievements of Biochemistry and Biophysics, Vol. 79, p. 355]
(1959)].

Example 1 Extraction and Isolation of Antioxidant Compounds 460 g of tamarind seed coat was crushed with a crusher (R-8 type, manufactured by Nippon Rikagaku Kikai Co., Ltd.), and 2 liters of ethyl acetate was added at 20 ° C. under normal pressure. Extracted 4 times with 40
Concentration under reduced pressure at or below ℃ gave 3.4 g of crude extract. The obtained crude extract was subjected to preparative high performance liquid chromatography [column: Novel Chemical Co., Ltd. Develosil ODS-10 (20 mm × 2
50 mm), solvent: 0.1% (w / v) trifluoroacetic acid-containing 17% (v / v) acetonitrile / water, flow rate:
5.0 ml / min, detection: 280 nm ultraviolet absorption] to separate an antioxidant activity fraction consisting of a single component and an antioxidant activity fraction consisting of a plurality of components. The solvent is removed under reduced pressure from the fraction consisting of a single component, and the antioxidant active substance 28
1.2 mg was obtained and was named compound 2.

On the other hand, the fraction containing a plurality of components was further subjected to preparative high performance liquid chromatography [column: Novel Chemical Co., Ltd. Develosil ODS-10 (20 mm × 250 mm),
Solvent: 0.1% (w / v) trifluoroacetic acid-containing 7%
(V / v) acetonitrile / water, flow rate: 5.0 ml /
Min., Detection: 280 nm ultraviolet absorption], and two fractions each containing a single component were collected. The solvent was removed from this under reduced pressure, and 21.3 mg and 7.8 of antioxidant active substances, respectively.
mg were obtained and were named compounds 1 and 3, respectively.

Example 2 Identification of Compounds 1 to 3 Structural analysis of Compounds 1 to 3 was carried out by ultraviolet absorption (UV) spectrum, infrared absorption (IR) spectrum, electron impact mass spectrometry spectrum (EI-MS), one-dimensional analysis. ¹ H nuclear magnetic resonance absorption ( ¹ H-NMR, CD ₃ OD, 400 MHz) and ¹³ C
Nuclear magnetic resonance absorption ( ¹³ C-NMR, CD ₃ OD, 100M
Hz). The results of Compound 1 are shown in Table 1, the results of Compound 2 are shown in Table 2, and the results of Compound 3 are shown in Table 3. Furthermore, two-dimensional NMR (COSY, H
MQC, HMBC), and from the results of Tables 1 to 3, compound 1 is methyl 3,4-dihydroxybenzoate represented by the above formula (2) and compound 2 is 3 represented by the above formula (3). , 4-dihydroxyphenylacetate, compound 3
To 2-hydroxy-3 ′, 4′- represented by the above formula (4)
It was identified as dihydroxyacetophenone.

[0021]

[Table 1]

[0022]

[Table 2]

[0023]

[Table 3]

Example 3 Antioxidant activity of compounds 1 to 3 0.13 ml of linoleic acid, 10 m of 99% distilled ethanol
The sample was added to the mixed solution of 1 to a final concentration of 20 μM, and 50 mM phosphate buffer (pH = 7.
0) 10 ml was added, and distilled water was added to make 25 ml. For comparison, a control to which no sample was added and a standard sample to which 20 μM of α-tocopherol was added were prepared. These were placed in a 50 ml stoppered flask, sealed tightly with Parafilm, stored at 40 ° C., and the antioxidant activity was measured with the Rhodan iron method and the TBA method over time. The results are shown in Fig. 1 (Rhodan iron method) and Fig. 2 (TBA
Method). In addition, the results are shown in% of the degree of oxidation in each sample addition system when the absorbance (oxidation degree) of the control (no addition) is 100%. Compounds 1 to 3 each showed an antioxidant activity at a concentration of 20 μM by both the rhodan iron method and the TBA method. Compounds 1 and 3 had strong antioxidant activity, and had the same level of antioxidant activity as α-tocopherol.

[0025]

As described above, according to the present invention,
Provided is a highly safe and highly active antioxidant derived from tamarind seed coat, which is suitable for foods, feeds, cosmetics, pharmaceuticals and raw materials thereof.

[Brief description of drawings]

FIG. 1 is a graph showing antioxidant activity measured by the Rhodan iron method.

FIG. 2 is a graph showing antioxidant activity measured by TBA method.

Claims (5)

[Claims] 1. Formula (1): (In the formula, R means a methoxycarbonyl group, an acetyl group or a hydroxymethylcarbonyl group), which is an antioxidant. 2. The antioxidant according to claim 1, wherein the compound of formula (1) is extracted from tamarind seed coat. 3. The antioxidant according to claim 1, which contains the compound of formula (1) as an ethyl acetate extract of tamarind seed coat. 4. Formula (3): The compound according to claim 4, which is represented by 5. Formula (4): embedded image The compound according to claim 4, which is represented by