JP3092297B2 - Method for producing natural antioxidants - Google Patents

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 added to foods, pharmaceuticals, cosmetics and the like.

[0002]

2. Description of the Related Art A thermoplastic resin such as a plastic film is susceptible to oxidative deterioration, so that mechanical strength is reduced, an odor is generated, and discoloration occurs. In addition, foods containing fats and oils are liable to be oxidized and deteriorated, not only making the appearance of the foods worse, but also accompanied by a deterioration in flavor and may further generate toxic substances.

As one of the measures, an antioxidant has been added. However, the widely used antioxidants BHT (butylhydroxytoluene) and BHA (butylhydroxyanisole), etc.
In recent years, involvement in carcinogenesis and adverse effects on the lungs and liver have been pointed out, and consumers are concerned about the safety of artificial synthetic chemicals, and interest in natural products has increased.

[0004] Antioxidants derived from natural products include tocopherols widely present in plants, flavone derivatives,
Coffee acid derivatives, gallic acid derivatives, gossypol, sezamole, and the like are known. On the other hand, from the viewpoint of plant types, complex antioxidants are obtained from spices, crude drugs, natural oils and fat plants, but no usable antioxidants have been obtained yet. In addition, a general method for extracting antioxidants from plant materials requires complicated procedures and treatments.

[0005]

SUMMARY OF THE INVENTION The present invention has been made in order to solve such problems, and an object of the present invention is to provide a method for easily extracting and separating antioxidants from available natural materials. Is to provide.

[0006]

In order to solve this problem, the present invention crushes and dries the leaves of Nanten (Nandina domestica Thunb.), Removes non-polar substances with a non-polar organic solvent, and removes the non-polar substances with a polar organic solvent. After extracting the polar substance and drying under reduced pressure, p
Adjusted to H3 to remove chloroform-soluble part, adjusted water-soluble part to pH 12 to extract chloroform-soluble part, dried again under reduced pressure, and obtained by preparative thin-layer chromatography. And a method for producing a natural antioxidant.

In the present invention, Nanten (Nandina domest)
ica Thunb.) can be easily obtained because it grows wild or planted in garden trees in the warm mountainous areas of Japan, and is used as a side dish for side dishes and the fruit is used as a crude drug (antitussive).

[0008] Nanten used in the present invention is selected from about 300 kinds of plant materials as a material having high antioxidant activity by antioxidant activity assay. The antioxidant activity was assayed by the following gravimetric method or thiocyanation method. FIG. 1 shows the results.

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

<Thiocyanation> A treatment solution having the following composition was prepared in a 50 ml Erlenmeyer flask, left in a thermostat at 40 ° C., and 0.1 ml each of the solution was taken out from the treatment day, and 75% ethanol was added. The mixture was placed in a wide-mouthed vial (32.7 ml capacity) containing 7 ml, and 0.1 ml of an aqueous solution of ammonium thiocyanate (30%) and 0.1 ml of ferrous chloride (12 × 10 ^-2 M, 3.5% aqueous hydrochloric acid) were added. 1 ml was added to develop the color, and 3 minutes later, the absorbance at 500 nm was measured.

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

[0012] In the figure, a blank refers to a sample containing no test solution. In addition, a commercially available BHT was treated in the same manner, and the activity was measured and compared.

Hereinafter, the present invention will be described in detail. As shown in FIG. 1, since the leaves of Nanten have strong activity, Nanten leaves are used in the present invention. Fresh or dried leaves are used for Nanten leaves. In the case of fresh leaves, grind with solvent as it is,
In the case of dry leaves, dry naturally in the shade or warm air (5
(Preferably 0 ° C. or lower) and pulverized before use.

Next, polar substances are extracted from the Nanten leaves. For efficient extraction, non-polar substances are removed in advance with a non-polar organic solvent such as hexane or dichloromethane, and the residue is subjected to antioxidation with a single polar organic solvent such as ethanol, methanol, acetone or a mixture of two or more solvents. Extract polar substances containing substances.

A basic 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, and then the chloroform-soluble portion is removed.
The remaining water-soluble part was adjusted to pH 12 with an aqueous solution of sodium hydroxide.
And a chloroform-soluble part is extracted, and this is dried under reduced pressure to obtain a basic substance containing an antioxidant.

The antioxidant is extracted from the extract.
The extract is banded on a thin-layer chromatographic plate and developed by a one-dimensional ascent method. The developing plate was checked for antioxidant activity with a linoleic acid ethanol solution and an N, N-dimethyl-p-phenylenediamine hydrochloride-ethanol solution, and the band where this activity was confirmed was scraped off and extracted with chloroform. Vacuum to dryness. Further, thin-layer chromatography is performed with a different solvent in the same manner as described above to separate the dye, extract and dry under reduced pressure. The substance thus obtained is used as it is or dissolved in an appropriate solvent, and used alone or mixed with other auxiliaries.

[0017]

EXAMPLE About 100 g of hot-air dried powder of Nanten leaves was extracted three times with 500 ml of hexane and three times with 500 ml of dichloromethane using a separatory funnel (all extraction operations were the same hereinafter). And then extracted three times with 99.5% ethanol. The extract was dried under reduced pressure using a rotary evaporator (hereinafter, the same applies to all dried under reduced pressure). This was dissolved in about 500 ml of distilled water and adjusted to pH 3 with sodium hydroxide or hydrochloric acid.
Extract several times with 1 of chloroform, discard it. The residue was further adjusted to pH 12, the same extraction was performed, the extract was dried under reduced pressure, and an appropriate amount was dissolved in chloroform.
It is used for the following thin layer chromatography.

[0018] This liquid as a thin layer chromatography on silica gel plates (Merck "60F _254"), chloroform: ether: acetic acid (parts by weight 92.2: 2.
3: 0.5 V / V) A one-dimensional ascent method using a solvent was performed.
An ethanol solution of 2.5% linoleic acid was sprayed on this developing plate, and irradiated with ultraviolet rays (365 μm) for 15 minutes.
When the 1% methyl-p-phenylenediamine hydrochloride-ethanol solution was sprayed, antioxidant activity was confirmed as a white spot on a pink fabric in a pF range of 0.4 to 0.5.

The portion where the antioxidant activity was confirmed was scraped into a band, extracted with chloroform, and dried under reduced pressure to obtain a light green solid. This was similarly subjected to a one-dimensional ascent method using a solvent of chloroform: methanol: ethyl acetate (96: 4: 0.5 by weight) in a thin-layer chromatograph, developed, and stripped of a portion other than the green dye in a strip shape. This was extracted with chloroform and dried under reduced pressure to give several mg of a separated component as a white powder.

It is confirmed by thin layer chromatography that the separated components are different from naturally occurring tocopherol and ascorbic acid.
The antioxidant activity was assayed in the same manner as in FIG. 1 using ml (equivalent to 0.5 mg of the separated component) as an assay solution, and also compared with commercially available ascorbic acid, δ-tocopherol, and BHT. Infrared spectra were also measured. The infrared spectrum is “FT / IR-5M” manufactured by JASCO Corporation.
P ". The results are shown in FIGS.

As is clear from FIG. 2, which is the result of assaying the antioxidant activity, the antioxidant activity of this separated component is stronger than δ-tocopherol, which is the strongest among tocopherols, and is slightly inferior to BHT. Do you get it. In addition, judging from FIG. 3, this separated component was estimated to be an aliphatic ester compound having an unsaturated bond.

[0022]

According to the present invention, an antioxidant can be separated from an ethanol extract of Nanten leaves, which has not been attempted to date, by a simple method called preparative thin-layer chromatography. It became. In addition, the isolated component according to the present invention is an antioxidant different from naturally-occurring tocopherols and ascorbic acid, has an activity superior to these, and has an activity close to BHT which has been widely used. Can be used as a substitute for

[0023]

[Brief description of the drawings]

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

FIG. 2 shows the results of assaying the components separated from Nanten leaves according to the present invention for the antioxidant activity by the thiocyanation method.
It is the graph shown compared with T and (delta) tocopherol.

FIG. 3 is a view showing an infrared spectrum of a component separated from Nanten leaves according to the present invention.

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int. Cl. ⁷ , DB name) C09K 15/34 BIOSIS (DIALOG) JICST file (JOIS)

Claims (1)

(57) [Claims] 1. A non-polar organic solvent is used to remove a non-polar substance from a leaf of Nanten (Nandina domestica Thunb.), A polar substance is extracted with a polar organic solvent, and then dried under reduced pressure. Remove the soluble part and adjust the water-soluble part to pH 12
A method for producing a natural antioxidant, comprising extracting a chloroform-soluble portion by adjusting the pressure, drying again under reduced pressure, and then obtaining the product by preparative thin-layer chromatography.