JPH0978061A - Antioxidant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antioxidant which is useful for preventing food products, feeding, cosmetics, medicines and the like from being deteriorated by oxidation by using a hydrolyzate of the extract from olive plant with a solvent as an active ingredient because of its excellent antioxidant activity originating from the natural product. SOLUTION: Methanol is added to olive leaves and they are heated at 60 deg.C for 15 minutes to effect extraction. This operation is repeated three times to collect the extract and the solvent is evaporated at 40 deg.C, freeze-dried to obtain an extract of olive leaves. This extract is mixed with 0.5N hydrochloric acid in 0.2% concentration, stirred at room temperature one overnight to give the hydrolyzate of the extract from olive leaf with a solvent. The hydrolyzate is adjusted in its pH to 3.0 with sodium hydroxide, subjected to column chromatography to collect the fractions eluted with 20% methanol aqueous solution. Then, the collected fraction is evaporated, freeze-dried to prepare the purified hydrolyzate of the extract from olive leaf. Finally, this purified hydrolyzate is used in food product, feedings, cosmetics, medicines and the like, as an active ingredient of a naturally occurring antioxidant.

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 increased 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. Under these circumstances,
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 vegetable oils, flavones contained in various plant tissues, quercetin, flavone derivatives such as rutin, tea leaves, and coffee. Beans, caffeic acid contained in cocoa 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. The present inventors have unexpectedly found that the hydrolyzate of olive plant solvent extract has an excellent antioxidant effect during various searches and studies on antioxidant components in natural products. Has been completed. It is known that solvent extracts of olive plants have antioxidant activity (Phytochemistry Vol.31, No.4, pp.1173-1178, 199).
2) It is also known that olive oil is an oil that is relatively difficult to oxidize, but it has never been known that the hydrolyzate of olive plant solvent extract has a very excellent antioxidant effect. Didn't.

[0004]

The present invention provides an antioxidant comprising a hydrolyzate of olive plant solvent extract as an active ingredient. The antioxidant of the present invention is derived from a natural product, has a high level of safety, and exhibits an excellent antioxidant effect, and is particularly suitable for use in foods, feeds, cosmetics, pharmaceuticals, etc. Is suitable.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION The olive plant used as the antioxidant of the present invention is an evergreen tree of the genus Olive, typically olive (Olea europaea) leaves, pulp, olive oil pomace, etc., or a mixture thereof. However, leaves and pulps are preferable because a component excellent in antioxidant action can be obtained by extraction and hydrolysis.

[0006] These are raw or appropriately dried or appropriately chopped, and water, a lower alcohol having 1 to 4 carbon atoms (eg, methanol, ethanol, propanol,
Extract with a solvent selected from butanol), acetone, ethyl acetate and a mixed solvent thereof. The extraction can be carried out using olive plants with an appropriate amount of solvent, and the extract is concentrated to obtain a crude extract. The extraction conditions are not particularly limited, but may be 8-16 at normal temperature to 30 ° C. under normal pressure to reduced pressure.
If the time is extracted 2 to 4 times, the extraction can be performed efficiently. Although a known method is used for concentration, freeze-drying or 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 hydrolysis. For hydrolysis, enzymes such as esterase,
Hydrolysis with acids such as hydrochloric acid, sulfuric acid and acetic acid can be adopted.
The hydrolysis conditions are also not particularly limited, but in the case of enzymatic hydrolysis, at an appropriate enzyme concentration, the optimum temperature of the enzyme to be used, pH, usually 30 to 55 ° C., pH 4 to 8
The hydrolysis is performed for about 0 to 24 hours. In the case of acid hydrolysis, hydrolysis is usually performed at room temperature to 60 ° C. for 2 to 24 hours at an acid concentration of 2 to 10% by weight. Although this has not been clearly demonstrated, this hydrolysis decomposes the polyphenol component in olive plants, resulting in hydroxytyrosol (JAOCS, Vol.68, No.9 (September), which has a stronger antioxidant power.
1991)), it is considered that the antioxidant activity is improved.

In the present invention, the hydrolyzate thus obtained may be used as it is as an antioxidant, and further purified by a method known per se, for example, chromatography, membrane separation, porous resin separation, and used. You may. Further, in the present invention, with these, known excipients acceptable for foods, cosmetics, pharmaceuticals and the like, and other suitable additives, for example, lactose, dextrin, gums, glyceric acid ester, etc., In solid or liquid form in a manner known per se, for example, powder, paste, emulsion, oil liquid,
It can also be used as a dosage form such as an aqueous solution. The amount of the hydrolyzate in such a formulation is appropriately selected depending on the intended use, the intended antioxidant effect, etc., but is usually 70
Up to 80% by weight, preferably 10-20% by weight.

[0009] The hydrolyzate of the present invention is obtained from olives whose fruits are edible, 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, it can be used by adding, mixing, moistening or other methods in the manufacturing process of foods and the like. The amount used can be appropriately selected according to the desired purpose, but normally, the concentration of the hydrolyzate is 0.0 in the desired product.
It should be 1 to 0.05% by weight. In particular, the antioxidant of the present invention exerts an excellent antioxidant action as an antioxidant for edible oil, and is also suitable as an antioxidant for oils and fats containing highly unsaturated fatty acids such as DHA.

It has also been found that the antioxidative effect is synergistically improved by the combined use with other antioxidizing agents such as tocopherol. Therefore, such combination is also within the scope of the present invention. The combination ratio can be appropriately selected, but in the case of tocopherol, the weight ratio of olive plant solvent extract hydrolyzate: tocopherol of the present invention is usually 1: 0.
When it is 5 to 1, a good synergistic effect is obtained.

[0011]

EXAMPLES Next, the present invention will be described in more detail by way of examples, which should not be construed as limiting the invention. Example 1 Methanol (1 liter) was added to olive leaves (100 g).
Was added and the operation of extracting at 60 ° C. for 15 minutes was repeated 3 times. The extracts were collected, the extraction solvent was evaporated at 40 ° C, and the extract was dried with a freeze dryer to obtain an olive leaf extract (36.6).
g) was obtained. To this extract (36.6 g), 0.5 N hydrochloric acid was added at a concentration of 0.2%, mixed and stirred overnight at room temperature to obtain a solvent extract hydrolyzate of olive leaf. After adjusting the hydrolyzate to pH 3.0 with sodium hydroxide, ODS
Column chromatography was performed using a column packed with (C ₁₈ ), 20% methanol aqueous solution eluate fractions were collected, and solvent evaporation and lyophilization were performed in the same manner as above to give a hydrolyzed purified product (7.0 g). ) Got.

Example 2 In the same manner as in Example 1, a hydrolyzed product (8.5 g) was obtained using olive pulp (100 g).

Example 3 The solvent extract hydrolyzed purified product of olive leaf obtained in Example 1 was added to the lard at a ratio of 0.01% by weight or 0.05% by weight and incubated at 70 ° C. The change in the peroxide value of lard with time was traced. Similarly, with respect to lard added with 0.1% by weight of a hydrolyzed and purified product of a solvent extract of olive pulp obtained in Example 2, the change in peroxide value with time was traced. As a control, lard containing 0.01% by weight of α-tocopherol and lard containing no antioxidant were similarly tested. The peroxide value (POV) was measured as follows.

Reagent solvent: a 2: 3 volume ratio mixture of chloroform: acetic acid. Saturated potassium iodide solution: Prepared by saturating 25 g of potassium iodide with 15 ml of distilled water. 0.01N sodium thiosulfate standard solution: 0.1N sodium thiosulfate standard solution was diluted 10 times with distilled water. 1% starch solution: 1 g of soluble starch in 100 parts of water
Prepared by filtering after adding to ml and heating to dissolve.

Measuring Method The fats and oils were accurately weighed, an antioxidant was added, and the mixture was incubated at a predetermined temperature. To this, 10 ml of the above solvent was added and dissolved, and the mixture was transferred to an Erlenmeyer flask with a stopper. 1 ml of saturated potassium iodide solution was added, stirred for 1 minute and left in the dark for 5 minutes. Then, 15 ml of water was added and after stirring for 1 minute, 1 ml of starch solution was added. Titration with 0.01N sodium thiosulfate standard solution was taken as the end point when the blue color due to starch disappeared.
g) was calculated.

[0016]

[Equation 1]

The results are shown in the graph of FIG. As is clear from FIG. 1, the olive plant solvent extract hydrolyzate, in particular, the olive leaf extract hydrolyzate markedly suppressed the production of peroxide in lard as compared with the control, and showed excellent anti-oxidant activity. It can be seen that it has an oxidizing action.

Example 4 Soybean oil was added with the olive leaf solvent extract hydrolyzate obtained in Example 1 in a proportion of 0.01% by weight, and as in Example 3, the peroxide value changed with time. Was tracked. As a control,
BHA 0.01 wt% added soybean oil, BHT 0.01 wt%
Similarly, the daily changes in the peroxide value of the added soybean oil and the antioxidant-free soybean oil were tracked. The results are shown in the graph of FIG. As is clear from FIG. 2, the hydrolyzate of olive leaf extract has an excellent antioxidant effect.

Example 5 To the lard was added 0.015% by weight of a 0.5: 1 weight ratio mixture of the olive leaf solvent extract hydrolyzate obtained in Example 1 and α-tocopherol, and the same as in Example 3. Then, the daily change of the peroxide value was traced. Also, the same applies to lard containing only 0.005% by weight of the olive leaf solvent extract hydrolyzate of Example 1, lard containing only 0.01% by weight of α-tocopherol, and lard without addition of an antioxidant. The daily change of oxide value was traced. The results are shown in the graph of FIG. As is clear from FIG. 3, when olive leaf solvent extract hydrolyzate and α-tocopherol are used in combination,
A significant improvement in the antioxidant effect due to the synergistic effect is seen.

Example 6 The olive leaf solvent extract hydrolyzate obtained in Example 1 was added to DHA at a ratio of 0.1% by weight, and the same as in Example 3, except that the incubation temperature was 37 ° C. hand,
The daily change in peroxide value was traced. As a control, BHA
0.1 wt% added DHA, catechin 0.1 wt% added DH
Similarly, the daily change of the peroxide value was traced for A and DHA without antioxidant. The results are shown in the graph of FIG. As is clear from FIG. 4, the hydrolyzate of olive leaf extract has an excellent antioxidant effect.

[0021]

As described above, according to the present invention,
Provided is an antioxidant derived from olive, which is highly safe and has excellent activity, which is suitable for foods, feeds, cosmetics, pharmaceuticals and raw materials thereof.

[Brief description of drawings]

FIG. 1 is a graph showing the antioxidant activity of olive plant solvent extract hydrolysates in lard.

FIG. 2 is a graph showing the antioxidant activity of olive leaf solvent extract hydrolyzate in soybean oil.

FIG. 3 is a graph showing the antioxidant activity of a combination of olive leaf solvent extract hydrolyzate and α-tocopherol in lard.

FIG. 4 is a graph showing the antioxidant activity of olive leaf solvent extract hydrolyzate in DHA.

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Claims (7)

[Claims] 1. An antioxidant comprising a hydrolyzate of olive plant solvent extract as an active ingredient. 2. The antioxidant according to claim 1, wherein the olive plant solvent extract is an extract of olive leaves with a solvent selected from water, lower alcohols, acetone, ethyl acetate and a mixed solvent thereof. 3. The antioxidant according to claim 1, wherein the olive plant solvent extract is an extract of olive pulp with a solvent selected from water, lower alcohols, acetone, ethyl acetate and a mixed solvent thereof. 4. The antioxidant according to claim 1, wherein the hydrolyzate is an enzymatic hydrolyzate or an acid hydrolyzate. 5. An antioxidant for edible oils.
The antioxidant according to any one of 1. 6. The antioxidant according to any one of claims 1 to 4, which is used in combination with another antioxidant. 7. The antioxidant according to claim 6, wherein the other antioxidant is tocopherol.