JPH06212154A - Production of oxidation-resistant substance - Google Patents

Abstract

PURPOSE:To obtain an-oxidation resistant substance which is suitable for preventing the quality degradation of foods contg. an oil, a fat, or a lipid by extracting the substance contained in a plant of the genus Quphea with a polar org. solvent. CONSTITUTION:A wild species of the genus Quphea originally coming from Mexico (e.g. quphea labia or Quphea ritei), pref. its seed, is washed with water, air-dried, crushed, and extracted for about 1-3hr by immersing it in a polar org. solvent (e.g. methanol). The extract is partially purified by adsorption chromatography, etc., fractionated by eluting with polar org. solvents with various concns., concentrated, and purified by reversed-phase chromatography, giving the objective substance. A crude product of the substance may be used in some applications.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a method for producing an antioxidant, and more particularly to a method for producing an antioxidant derived from a natural product extracted and collected from a plant of the genus Cuphea.

[0002]

BACKGROUND OF THE INVENTION The main cause of chemical deterioration of food is due to lipid oxidation. In particular, foods containing unsaturated fatty acids are easily oxidized,
It causes a decrease in flavor and develops toxicity. Also,
In the human body, various radical species produced by lipid peroxidation cause DNA damage and mutation,
It has been clarified that it is greatly involved in cancer and aging.

From such a background, a substance that suppresses oxidation in food and in vivo has been attracting attention. Antioxidants have come into widespread use for the purpose of preventing the oxidation of foods because the synthetic antioxidants butylhydroxyanisole (BHA) and dibutylhydroxytoluene (BHT) have been developed since the 1950s. Because.

By the way, in recent years, safety has come to be questioned because it is pointed out that these substances are carcinogenic and have an adverse effect on the lungs and liver.
On the other hand, there is a tendency toward natural products by consumers, and the amount of use is drastically decreasing. Recently, tocopherol, which is a natural antioxidant, has been added to many foods in place of these synthetic antioxidants. Tocopherol is contained in many agricultural crops and there is no problem in terms of safety, but the essential antioxidant power is not sufficient, and the development of stronger natural antioxidants is desired. It is also known that flavonoid compounds contained in plants have antioxidant properties.

The present inventors searched for antioxidant substances contained in plants of the genus Cuphea, which have been attracting attention in recent years, by producing medium-chain fatty acid triglyceride (MCT). As a result, the plant of the genus Cuphea has antioxidant properties. They have found that they are producing substances, and have completed the present invention based on these findings.

[0006]

That is, the present invention relates to a method for producing an antioxidant, which comprises extracting and collecting an antioxidant contained in a plant of the genus Cuphea with a polar organic solvent.

[0007] The plant of the genus Cuphea used in the present invention is a wild species whose origin is Mexico, and studies on cropping have been conducted in the United States and the like. As a Kufea plant,
Although it is not particularly limited and can be arbitrarily used, cuphea ravia, cuphea retainer, cuphea laminudilina, cuphea tolcana, cuphea reputapoda and the like are particularly preferable. Also, regarding the applied part, the seed,
There are leaves, stems, etc., and although not particularly limited, seeds are usually used.

Examples of polar organic solvents used for extracting antioxidants from plants of the genus Cuphea include methanol, ethanol, acetone and the like, and these solvents are usually used in a mixed state with water.

Explaining the details of the method for producing an antioxidant according to the present invention, seeds of the genus Cuphea are washed with water, air-dried, crushed and then immersed in a polar organic solvent for extraction. In addition, MC
When impurities such as T and impurities are contained, they are treated with a solvent such as hexane for removal, and then extracted. The extraction time is not particularly limited, but is usually about 1 to 3 hours. When the polar organic solvent is used as a mixed liquid with water, the compounding ratio is preferably the former: the latter = 40 to 100: 60 to 0. The extraction operation can be repeated several times as needed.

Purification and concentration of the extract can be carried out by a conventional method such as various kinds of chromatography. As a specific method, after partial purification by adsorption chromatography or the like, fractionation and concentration are carried out by eluting with a polar organic solvent of various concentrations. In addition, reverse phase chromatography can be applied to increase the degree of purification. For example, it is possible to fractionate into a fraction having an antioxidant property by high performance liquid chromatography using YMC-pack polymer C18. The antioxidant substance obtained by the present invention may be not only a purified product but also a crude product depending on its application.

The antioxidative activity may be measured by a conventional method. As one example, as shown in Table 1, in a system in which ethyl linoleate is forcibly peroxidized with Fenton's reagent, cuphea is used. It can be performed by adding an extract from a seed of a genus plant or a control (25 μM α-tocopherol or BHT), reacting at 35 ° C. for 16 hours, and measuring the produced malonaldehyde by the thiobarbituric acid (TBA) method. . The TBA method is carried out by heating a solution having the composition shown in Table 2 for 1 hour in boiling water, adding butanol to develop the color, and measuring the absorbance at 532 nm.

[0012]

[Table 1] Table 1 Ethyl linoleate 10 μl Tris-hydrochloric acid buffer (0.05 M, pH 7.4) containing 0.2% SDS and 0.15 M KCl 5 ml 20 mM FeCl ₂ 50 μl 10 mM H ₂ O ₂ 50 μl Cuphea plant Extract or control ethanol solution 100 μl

[0013]

[Table 2] Table 2 Fenton reagent reaction solution 0.2 ml 4% BHT ethanol solution 50 μl 8% SDS solution 0.2 ml 0.67% TBA solution 1.5 ml Acetate buffer (1N, pH 3.5) 1.5 ml

When the antioxidant properties of five kinds of Qupheea plant seed extracts were examined by the above-mentioned method, all of them had antioxidant properties as shown in FIG. Also,
Extracts such as leaves and stems are about 1/2 to 1 of the seed extract.
It had an antioxidant power of / 3. Most of these Qupheea plant extracts are tasteless and odorless and do not adversely affect the flavor when added to foods. Some of them, like Kufeea leptapoda, have a coconut-like aroma, and this property can also be used positively.

The antioxidant contained in the Qufeea plant can be purified by various chromatographies. For example, the antioxidant contained in the seed of Qupheier ravia is MCT with hexane as shown in FIG. After removing impurities and impurities, extraction with a polar organic solvent, adsorption on a resin packed in a column (Amberlite XAD-2, etc.), and elution with water and various concentrations of methanol and acetone can be performed for fractionation. FIG. 3 shows the antioxidant activity of 1 mg of each fraction. By eluting with various concentrations of aqueous methanol solution, it can be concentrated about 10 times.

Since flavonoids are well known as antioxidant substances contained in plants, 14 fractions having antioxidant properties obtained by the method of the present invention will be used.
Modified avis method (Toshiro Nakabayashi, Journal of Food Industry Society, vol. 9, 28
Pp. 38, 1962).
As shown in the table, there are 4 fractions that are considered to belong to flavonoids, and 5 are those that cannot be judged.
Five fractions were determined to be non-flavonoids. Since these substances were obtained by removing the hexane extract, it is clear that they are not tocopherols. In addition, the hexane extract did not show any antioxidant property.

[0017]

[Table 3]

As described above, the antioxidant substances obtained by the present invention include not only tocopherol but also those belonging to flavonoids and other substances, and they may be novel substances. It is also possible to obtain an antioxidant substance from the residue after extracting MCT with hexane.

[0019]

EXAMPLES Next, the present invention will be described in detail with reference to Examples. Example 1 19 g of cuphea larvia seeds were washed with water, air-dried, crushed, immersed in a 3-fold amount of 80% ethanol-water mixture overnight, and extracted twice with the same solvent. The extract was concentrated under reduced pressure and dried to obtain 1.76 g of extract. next,
100 μl of linoleic acid and 40 μg of 80% ethanol extract of the seeds of cuphea larvia described above were used, and the inner diameter was 1.6 m.
It was put in a test tube having a length of m and a length of 12 cm, the solvent was removed under nitrogen gas, and the oxidation was measured at 30 ° C. in the dark. On the other hand, as a control, an 80% ethanol-water mixed solution was added instead of the cuphea seed extract. Oxidation was measured by the Rodan iron method. That is, 4.8 ml of 75% ethanol, 0.1 ml of 30% ammonium rhodanide and 0.1 ml were added to the sample.
Add 20 ml of ferrous chloride (20 mM), react for 3 minutes, then add 50
Absorbance at 0 nm was measured. The results are shown in Table 4.

[0020]

[Table 4] Table 4 ───────────────────────── Samples Increase in absorbance 1 day 3 days ────────── ————————————————————————————————— Cofe genus plant seed extract 0.0461 Unadded 0.041 0.218 ————————————————— ──────────

As is clear from the table, the addition of cuphea seed extract suppressed the oxidation of linoleic acid.

Example 2 Using the Cufaea seed extract obtained in Example 1, the oxidative stability of soybean oil was tested in the same manner as in Example 1. The results are shown in Fig. 4. The addition of cuphea seed extract suppressed the oxidation of soybean oil.

[0023]

INDUSTRIAL APPLICABILITY According to the present invention, a substance rich in antioxidant can be obtained from a plant of the genus Cuphea. It is possible to suppress the deterioration of quality by adding this antioxidant substance to foods containing fats and oils and lipids.

[Brief description of drawings]

FIG. 1 is a graph showing the antioxidant properties of 80% ethanolic extracts of cuphea seeds and control substances.

FIG. 2 shows a method for fractionating antioxidants contained in seeds of the genus Cuphea.

FIG. 3 is a graph comparing the antioxidant properties of antioxidants extracted from cuphea seeds and control substances against the oxidation of linoleic acid ethyl ester.

FIG. 4 is a graph comparing the antioxidant properties of antioxidants extracted from seeds of the genus Cuphea and control substances against the oxidation of soybean oil.

Front page continuation (72) Yoshiyuki Fujii, 1-20-46, Zao-cho, Fukuyama City, Hiroshima Prefecture (72) Naoki Hosoi, 269, Kamiyama Minami, Numakuma-cho, Numakuma-gun, Hiroshima Prefecture (72) Kazumi Tachikada, Tsukuba, Ibaraki Prefecture 1-chome, Kasuga, Ichi 4203 Building-308

Claims (3)

[Claims] 1. A method for producing an antioxidant, which comprises extracting and collecting an antioxidant contained in a plant of the genus Cuphea with a polar organic solvent. 2. The production method according to claim 1, wherein seeds of the plant are used as a plant of the genus Cuphea. 3. The process according to claim 1, wherein the polar organic solvent is selected from methanol, ethanol and acetone.