JPH06108049A - Production of natural antioxidant - Google Patents

Abstract

PURPOSE:To provide a method to easily extract and separate an antioxidant from an easily available natural material. CONSTITUTION:Leaves of a Japanese snowbell (Styrax japonica Sieb. et Zucc.) are dried, ground, treated with a nonpolar organic solvent to remove nonpolar substances, treated with a polar organic solvent to extract polar substances, and evaporated to dryness in a vacuum, and the pH of the dried product is adjusted to 3 to obtain chloroform-extractibles. A saturated sodium hydrogencarbonate solution is added to the extractibles to remove chloroform- solubles, and the pH of the remaining water-solubles is adjusted to 3 to obtain a chloroform-solubles, which are recovered by split-sampling high-performance liquid chromatography.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a thermoplastic resin in industrial products or an antioxidant substance added to foods, pharmaceuticals, cosmetics and the like.

[0002]

2. Description of the Related Art Since a thermoplastic resin such as a plastic film is susceptible to oxidative deterioration, mechanical strength is reduced, an offensive odor is generated, and discoloration occurs. Further, foods containing oils and fats are also susceptible to oxidative deterioration, which not only makes the foods look unattractive but also deteriorates the flavor, and may also produce toxic substances.

As one of the measures against this, an antioxidant is added. However, antioxidants such as BHT (butylhydroxytoluene) and BHA (butylhydroxyanisole), which were once widely used,
In recent years, the involvement in carcinogenesis and the adverse effects on the lungs and liver have been pointed out, and consumers are concerned about the safety of artificial synthetic chemicals.

Antioxidants derived from natural products include tocopherols widely present in plants, flavone derivatives,
Caffeic acid derivatives, gallic acid derivatives, gossypol, sezamol, etc. are known. On the other hand, from the viewpoint of plant type, although complex antioxidants are obtained from spices, crude drugs, natural oil and fat plants, etc., usable antioxidants have not yet been obtained. Further, a general method for extracting an antioxidant from a plant material requires a troublesome procedure and treatment.

[0005]

SUMMARY OF THE INVENTION The present invention has been made to solve such a problem, and its object is a method for easily extracting and separating an antioxidant from an available natural material. Is to serve as a bank.

[0006]

Means for Solving the Problems In order to solve this problem, the present invention is to dry and grind the leaves of Egonaceae, remove nonpolar substances with nonpolar organic solvents, and extract polar substances with polar organic solvents. After drying under reduced pressure, the pH is adjusted to 3 to obtain a chloroform extraction part. Then, a saturated aqueous solution of sodium hydrogen carbonate was added to this to remove the chloroform-soluble part, and the remaining water-soluble part was further adjusted to pH 3 to obtain a chloroform-soluble part, which was obtained by preparative high performance liquid chromatography. To provide a method for producing a natural antioxidant substance characterized by:

[0007] In the present invention, the Japanese cypress is a deciduous small tree that is widely distributed in the temperate zone of Hokkaido to Okinawa in the subtropical zone in Japan, and it is also used for landscaping, and is easily available. On the other hand, the extract of Perilla frutescens is recognized as a preservative for foods.

The Egonoki tree used in the present invention was selected from about 300 kinds of plant materials as a material having a high antioxidant activity by an antioxidant activity assay. The antioxidant activity was assayed by the following gravimetric method or thiocyanation method. The results are shown in FIG.

<Gravimetric method> linoleic acid 5 purified by distillation
Take 00mg in a vial (32.4ml capacity),
After adding 0.1 ml of the extract and stirring well and removing the solvent under reduced pressure, place it in a thermo-hygrostat at 40 ° C and RH 50%.
Thereafter, the weight was measured every day for 4 to 5 days.

<Thiocyanation method> A treatment liquid having the following composition was prepared in a 50 ml Erlenmeyer flask, left in a thermostat at 40 ° C., and 0.1 ml each was taken out from the treatment day, 7
The mixture was placed in a wide-mouth vial (32.7 ml capacity) containing 5% ethanol (9.7 ml), and further 0.1 ml of an aqueous solution of ammonium thiocyanate (30%) and ferrous chloride (1).
0.1 ml of 2 × 10 ^-2 M, 3.5% hydrochloric acid aqueous solution) was added to develop color, and after 3 minutes, the absorbance at 500 nm was measured.

<Composition of treatment liquid> Ethanol (99.5%) 1.90 ml Linoleic acid ethanol liquid (2.5%) 2.05 ml Phosphate buffer solution (0.05 M, pH 7.0) 4.00 ml Distilled water 1.95 ml Assay solution (ethanol extract) 0.10 ml Total 10.00 ml

[0012] In the figure, the blank refers to a blank containing no assay solution. Further, a commercially available product of BHT was treated in the same manner, and the activity was measured and compared.

The present invention will be described in detail below. As shown in FIG. 1, since the leaves of Egon tree have a strong activity, the leaves of Egon tree are used in the present invention. As the leaves of the echinoki, raw leaves or dried leaves are used. In the case of fresh leaves, they are ground with a solvent as they are, and in the case of dried leaves, they are either naturally dried in the shade or dried with warm air (50 ° C or lower), and then crushed before use.

Then, a polar substance is extracted from the leaves of the echinacea tree. To promote extraction efficiently, remove non-polar substances in advance with non-polar organic solvents such as hexane and dichloromethane, and use antioxidants with the polar organic solvents such as ethanol, methanol, and acetone alone or in a mixed solvent of two or more types. Extract polar substances containing substances.

An acidic substance is extracted from this extract. The extract is dissolved in water, adjusted to pH 3 with an aqueous solution of sodium hydroxide or hydrochloric acid, the chloroform-soluble portion is taken, dried under reduced pressure, and dissolved in a saturated aqueous solution of sodium hydrogen carbonate. After this, the chloroform-soluble part was removed and the water-soluble part was adjusted to pH.
After adjusting to 3, an acidic substance in the chloroform-soluble part is obtained.

Antioxidants are separated from this extract. The extract was dissolved in ethanol and subjected to preparative high performance liquid chromatography using methanol as an eluent to give a concentration of 20-
Fractionation is carried out stepwise in the range of 80%. As shown in the results of FIG. 3, a fraction having a strong antioxidant activity is obtained in the range of 65 to 75%. Further, this fraction is fractionated with an eluent of 50% methanol. As shown in the results of FIG. 4, two components A and B are obtained, which are dried under reduced pressure. The substance thus obtained is used as it is, or dissolved in a suitable solvent, and used alone or mixed with other auxiliary materials.

[0017]

[Examples] 1.7 kg of dried hot air dried powder of Egonoki was extracted once with 2.0 l of hexane and three times with 2.0 l of dichloromethane in this order using a separating funnel (all extraction operations are the same below). Then, 2.0 l of 99.5% ethanol was added to this residue and the mixture was extracted 3 times, and the extract was dried under reduced pressure using a rotary evaporator (the same applies to reduced pressure dryness hereafter).
The ethanol-soluble portion was dissolved in 1.0 liter of distilled water and adjusted to pH 3 with sodium hydroxide or hydrochloric acid, and then 2
Extract several times with 00 ml of chloroform, combine the extracts, and dry under reduced pressure. A saturated aqueous solution of sodium hydrogencarbonate is added to and dissolved in this, and extracted several times with chloroform to remove it. The aqueous solution of this residue is adjusted to pH 3, extracted with chloroform several times, and dried under reduced pressure. By the above operation, 1.5 g of acidic substance was obtained.

An acidic substance is dissolved in 99.5% ethanol and preparative high performance liquid chromatography (manufactured by JASCO Corporation: "Chromatograph LC80" using methanol as an eluent).
0 series ”) (Used column: ODS-10, 30 × 2
50 mm (manufactured by Nomura Chemical Co., Ltd .: “DEVELOSI
L ”)), and the antioxidant is separated at a flow rate of 10 ml / min. That is, first, a fraction in the range of 65 to 75% is divided in the stepwise fractionation with a methanol concentration of 20% to 80%, and then this is fractionated with an eluent of 50% to obtain two components A and B. To collect. Both components are dried under reduced pressure. Two
The component yields were about 200 mg A and about 100 mg B.

The two components were dissolved in ethanol and assayed in the same manner as in FIG. 1 while comparing with commercially available BHT and δ-tocopherol. The assay results of antioxidant activity are shown in FIG. It was found that both of the two components are much stronger than δ-tocopherol, which has the highest activity among the tocopherols, and are superior to BHT.

[0020]

INDUSTRIAL APPLICABILITY According to the present invention, it has become possible to easily separate the antioxidant from the ethanol extract of the leaves of Egonaceae, which has never been tried to be used, by preparative high performance liquid chromatography. . Moreover, since it was found that the separated component according to the present invention has a higher activity than tocopherols and BHT, it can be used as a substitute for BHT.

[0021]

[Brief description of drawings]

FIG. 1 is a graph showing the results of assaying the antioxidant activity of an ethanol extract of leaves or fruits of Egonaceae by a gravimetric method and a thiocyanation method.

FIG. 2 is a graph showing the results of assaying the antioxidant activity of the component isolated from the leaves of Scutellaria barbata according to the present invention by the gravimetric method and the thiocyanation method, in comparison with BHT and δ-tocophenol.

FIG. 3 is a graph showing the results of stepwise fractionation of acidic components fractionated from the leaves of Egonaceae according to the present invention by preparative high performance liquid chromatography.

FIG. 4 is a graph showing the results of further fractionation of acidic components fractionated from the leaves of Egonaceae according to the present invention, which were fractionally fractionated by preparative high performance liquid chromatography.

Claims (2)

[Claims] 1. A goat tree (Styrax japonica Sieb. Et Zuc)
c.) A method for producing a natural antioxidant, which is obtained from leaves. 2. A cypress (Styrax japonica Sieb. Et Zuc)
c.) Leaves are treated with non-polar organic solvents to remove non-polar substances, polar organic solvents are used to extract polar substances, dried under reduced pressure, adjusted to pH 3 and the chloroform-soluble portion is collected. A natural antioxidant substance characterized by being obtained by preparative high performance liquid chromatography using a polar organic solvent as an eluent to extract a chloroform-soluble portion by adjusting the pH of the water-soluble portion obtained by adding an aqueous solution Production method.