JPH07241181A - Method for stabilizing food or the like containing colorant and/or vitamin - Google Patents

Abstract

PURPOSE:To prevent a colorant or vitamins from destroying without changing the color tone by adding a solubilized hesperidin to a food, etc., containing the colorant and/or vitamins. CONSTITUTION:This method for stabilizing a food, etc., containing a colorant and/or vitamins is to add about 0.01-1wt.% solubilized hesperidin to a liquid, a gelatinous or a solid food and drink, a cosmetic, a medicine, etc., containing the colorant and/or vitamins (e.g. vitamin A or B group, C or D) dissolved therein. Thereby, ultraviolet rays of sunlight, a fluorescent lamp, etc., are absorbed to prevent the colorant and/or vitamins poor in stability from destroying. Similar effects are obtained even by adding solubilized naringin in place of the solubilized hesperidin thereto.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a method for stabilizing colorants and / or vitamins.

[0002]

2. Description of the Related Art Conventionally, flavonoids such as hesperidin, rutin, and naringin are known to stabilize colorants and vitamins due to their strong UV-absorbing action in addition to their medicinal effects. ing. However, since these flavonoids are poorly soluble in water, the amount that can be added to the target is limited. The inventors of the present invention have a technology for glycosidating these flavonoids to efficiently produce solubilized flavonoids (Patent Application No. 5-25614).
No. 1) was developed.

Of these solubilized flavonoids, α-
Since glucosyl rutin and α-glucosyl quercetin are yellow, the color tone changes when added to an object. Therefore, the amount that can be added to the object is limited.

[0004]

The solubilized hesperidin used in the present invention is mainly a compound in which glucose is bound to the 4-position of glucose of hesperidin by an α-1,4 bond (hereinafter, this compound is referred to as “α-glucose”). Pyranosyl hesperidin ") and its glucose at the 4-position with 1 to 10 or more α-1,4 bonds of glucose.

The solubilized naringin used in the present invention contains glucose α-1, -1, at the 4-position of glucose of naringin.
A compound bound by 4 bonds (hereinafter, this compound is referred to as "α-glucopyranosyl naringin") and glucose at the 4-position of glucose with one α-1,4 bond to 1 ~.
It is a mixture mainly composed of 10 or more bonded together.

The object of the present invention has a colorant (preferably 0.01 to 1% by weight based on the object) and / or a vitamin (preferably 0.01 to 1% by weight based on the object) dissolved therein. Liquid, gel or solid food and drink products, cosmetics, pharmaceutical products and the like (hereinafter, these are collectively referred to as food and the like).

Coloring agents include crocin (carotenoid pigment), phycocyanin (porphyrin pigment), beet red (betacyanine pigment), safflower yellow pigment (flavonoid pigment), annatto pigment (carotenoid pigment) and cochineal (anthraquinone pigment). ) Etc. are not particularly limited, but those with poor stability are targeted.

Vitamins are ordinary vitamins, for example, vitamins A, B, C and D.

The present invention is particularly effective when the object is contained in a transparent or translucent container and is exposed to sunlight or fluorescent light. The amount of solubilized hesperidin or solubilized naringin used is usually 0.01 to 1% by weight from the viewpoint of effectiveness. The solubilized hesperidin or the solubilized naringin is made into a solution having a high concentration and mixed or dispersed in the object.

The solubilized hesperidin or solubilized naringin is CGTavse, that is, 1,4-α-D-gluca.
n; 4-α-D- (1,4-glucano) -tra
nsferase (EC 2.4.1.19.) can be used for production (Patent Application 5-25
6141). Any enzyme may be used as long as it is CGTase. However, since the yield of solubilized hesperidin or solubilized naringin is remarkably reduced when a substance which is unstable in an alkaline region is used, an alkali-resistant substance is preferably used.

The Bacillus strain A2-5a developed by the inventors of the present application (Deposit F of the Institute of Biotechnology, Institute of Industrial Science and Technology)
The novel CGTase collected from the culture of ERM P-13864 (hereinafter referred to as the present strain) retains high activity in the alkaline region and is therefore preferably used. The culture conditions of this strain and the method of collecting the enzyme are not special.

The conditions for sugar transfer to hesperidin or naringin using this novel CGTase are the same as in the conventional method except that they are carried out in the alkaline region, and they are not special. Preferably pH 8-11, temperature 20-
75 ° C, the concentration of the donor is 0.1 to 30%, preferably 1 to
20%, the concentration of hesperidin or 0.1-5%, preferably 0.5-2%, the enzyme 0.1-100 units /
Start the reaction to make ml. The longer the reaction time, the better as long as the enzyme activity is maintained. 100 reactions
Stop by heating at 5 ° C for 5 minutes.

As the donor, various oligosaccharides such as cyclodextrin and maltooligosaccharide, and α-1,4-glucans having α-1,4 bond such as starch are used.

As an example of the above means, solubilized hesperidin was produced as follows and then isolated to determine its structure. 0.5% hesperidin, 5% soluble starch, 1,
600 m of reaction solution containing 200 units of CGTase
1 was reacted at 40 ° C. for 16 hours. After the reaction, the reaction was stopped by heating at 100 ° C. for 5 minutes. To this, 10 units of β-amylase was added, reacted at 40 ° C. for 1 hour, and
The reaction was stopped by heating at 00 ° C for 5 minutes. This was applied to an Amberlite XAD-16 column, the column was washed with distilled water, and then Amberlite XAD- with 50% ethanol.
The hesperidin glycoside adsorbed on 16 was eluted. This eluate was concentrated and purified by preparative FPLC (column; ODS, solvent; 20% ethanol, flow rate; 2 ml / min). Further, this is prepared by TLC (silica gel plate, developing solvent; chloroform: methanol: distilled water =
65:35:10 upper layer, developed by ascending method), scrape the spots having absorption at 253 nm from this silica gel plate, extract with distilled water, freeze-dry these,
The main reaction products were hesperidin monoglucoside and diglucoside 4.2.
mg, 15.4 mg were obtained.

The compounds believed to be hesperidin monoglucoside and diglucoside purified as described above are 10 units / ml of glucoamylase or α-.
Treatment with glucosidase at 40 ° C. for 16 hours produced hesperidin and glucose in a molar ratio of about 1: 1 and 1: 2, respectively. Moreover, it was not decomposed by β-glucosidase treatment under the same reaction conditions as these. From these facts, these compounds show that hesperidin is added to glucose at a molar ratio of 1: 1 and 1: 2, respectively.
It was found to be hesperidin monoglucoside and diglucoside linked at -1,4.

The hesperidin released by this enzyme treatment was analyzed by the following HPLC (ODS), and the released glucose was analyzed by the glucose oxidase method (I. Mi.
waetal. Clin. Chem. Acta , 37,
538-540 (1972)). HPLC (ODS) analysis conditions

Further, the structure of hesperidin monoglucoside was analyzed in detail by methylation analysis. Dissolve sufficiently dried hesperidin monoglucoside in 0.25 ml of dimethyl sulfoxide, add newly prepared methylsulfinyl carbanion,
The reaction was carried out at room temperature for 2 hours. To this, 0.15 ml of methyl iodide was added and reacted at room temperature for 4 hours for methylation. Methylated product in 90% (v / v) formic acid at 100 ° C. for 1 hour,
Then, it was hydrolyzed in 1N sulfuric acid at 200 ° C. for 4 hours.
Add 13 mg of NaBH ₄ to the methylated product at 100 ° C for 2
After reduction for an hour, the mixture was reacted in 0.5 ml of acetic anhydride / pyridine mixed solution (1: 1, v / v) at 100 ° C. for 1 hour for acetylation. The partially methylated alditol acetate prepared in this manner was analyzed by gas chromatography (GC). The analysis conditions of GC are as follows. As a standard sample, hesperidin and maltosyl-
β-cyclodextrin was used.

[0018]

[Table 1]

The results of the GC analysis are shown in Table 1. From hesperidin monoglucoside 2,3,4,6-te
tra-O-methyl-glucitol ace
tate, 2,3-di-O-methyl-gluc
itol acetate, 2,3,6-tri-O-
Since methyl-rhamnitol acetate was detected, the glucose transferred by CGTase was α- at the 4-position of hesperidin glucose.
It was revealed that they are linked by 1,4 bonds, that is, α-glucopyranosyl hesperidin.

FIG. 2 shows a HPLC (ODS) chromatograph (analysis conditions are as described above) when the hesperidin glycoside was treated with β-amylase.
From this, it is considered that a series of hesperidin glycosides has a sugar bound to hesperidin in maltose units.

From the results of these enzymatic treatments and methylation analysis, solubilized hesperidin was mainly found in α-glucopyranosyl hesperidin and its glucose at the 4-position, and several to ten or more glucose atoms with α-1,4 bonds. It was found to be a mixture of individual bonds.

Similarly, α-glucopyranosyl naringin can be isolated from solubilized naringin.

[0023]

The reaction product obtained by the above production method, that is, the absorption spectrum of solubilized hesperidin is investigated,
It was compared with the absorption spectrum of solubilized rutin as a control (FIG. 1). As shown in FIG. 1, solubilized hesperidin has a strong absorption in the ultraviolet region. Also, 350-400 nm
It was found that there was no absorption in the vicinity and it was extremely colorless.
On the other hand, solubilized rutin showed strong absorption at that wavelength. That is, it has a deep yellow color. The solubility of α-glucosyl hesperidin (monoglucoside) or diglucoside in water was improved by about 350 or 420 times that of hesperidin. The molar extinction coefficient of α-glucosyl hesperidin (monoglucoside) or diglucoside was 8900 and 6800, respectively.

(Comparative Example 1) 0.01%, 0.05% and 0.1% (wt%) of solubilized hesperidin was added to a solution containing 0.05% (wt%) of phycocyanin (porphyrin dye), Put in a test tube and test section 1, 2, 3 respectively
And At the same time, 0.01%, 0.05%, 0.1
% (Wt%) of solubilized rutin was added to the test sections 4, 5 and 6, respectively. Let these stand at 4 ℃, 12,
Irradiation from all sides with a fluorescent lamp of 000 lux, the absorbance was measured at the maximum absorption of phycocyanin at 620 nm over time, and the absorbance when starting to stand (0.6 in this case)
Was set to 100, and the residual amount of the dye was shown as a percentage. At this time, a phycocyanin solution containing neither solubilized hesperidin nor solubilized rutin was used as a control. The control area gradually faded (absorbance decreased) over time, and
After 0 hours and 100 hours, the residual amount of dye is 3 respectively.
It became 0% and 20%. On the other hand, the dye in the test section was stabilized, and the residual amount of the dye after 50 hours was in the test sections 1, 2, 3
Was 65%, 75%, and 80%, respectively. In the test plots 2 and 3, the initial color tone was visually maintained. Moreover, after 100 hours, they were 45%, 55%, and 60%, respectively. On the other hand, a test section added with solubilized rutin, especially,
In the test groups 5 and 6, the blue color of the phycocyanin was changed to green because the solubilized rutin had a yellow color tone.

[0025]

【Example】

(Example 1) 0.01%, 0.05% and 0.1% (wt%) of solubilized hesperidin was added to a solution containing 0.05% (wt%) of gardenia crocin (carotenoid pigment). , Put in the test tube and test section 1, 2, 3 respectively
And Let these stand at 4 ℃, 12,000lux
Was irradiated from all sides with a fluorescent lamp of No. 4, and the absorbance was measured at the maximum absorption of crocin at 442 nm with time. The absorbance at the start of standing was set to 100, and the residual dye amount was shown as a percentage. At this time, a crocin solution containing no solubilized hesperidin was used as a control. The control group gradually faded over time (absorbance decreased), and
After 8 hours, the residual amounts of dye were 25% and 5%, respectively. In comparison, in the test plots, the dye was stabilized, and the residual amount of dye after 4 hours was 45 in test plots 1, 2 and 3, respectively.
%, 60% and 75%. In the test plot 3, the initial color tone was visually maintained. After 8 hours, the rates were 10%, 25%, and 50%, respectively.

(Example 2) 0.005% (wt%) of riboflavin, which is a vitamin B ₂ and a yellow pigment,
Solubilized hesperidin 0.01%, 0.05
%, 0.1% (wt%), and put into a test tube to make test sections 1, 2, and 3, respectively. Let them stand at 4 ° C for 1
Irradiation was carried out from all sides with a 2,000 lux fluorescent lamp, and the absorbance was measured at the maximum absorption of riboflavin at 445 nm over time. The absorbance at the start of standing was set to 100, and the residual amount of riboflavin was shown as a percentage. At this time, a riboflavin solution containing no solubilized hesperidin was used as a control. The control section rapidly faded (absorbance decreased) with the passage of time, and the residual amount was 10 after 2 hours.
Fell below%. In comparison, riboflavin in the test plots was stabilized, and the residual amounts after 2 hours were 45%, 60%, and 70% in the test plots 1, 2, and 3, respectively. Moreover, the residual amount of the test section 3 was 50% or more even after 4 hours.

Example 3 Solubilized hesperidin was added at 0.01%, 0.05% and 0.1% (% by weight) to a solution containing 0.1% (% by weight) of cochineal (anthraquinone dye), Test tubes 1, 2 and 3 were placed in test tubes. These were allowed to stand at 4 ° C., irradiated from all sides with a 12,000 lux fluorescent lamp, the absorbance was measured at the maximum absorption of cochineal at 525 nm, and the absorbance at the start of standing was set to 100, and the residual dye amount was determined. Is shown as a percentage. At this time, a cochineal solution containing no solubilized hesperidin was used as a control. The control section gradually faded over time (absorbance decreased), and the
After the lapse of time, the residual amounts of the dyes became 20% and 10%, respectively. In comparison, the test area was stabilized with the dye and 10
The remaining amount of dye after 25 hours was 25 in each of test sections 1, 2 and 3.
%, 50% and 75%. Further, the residual amount of the dye in the test section 3 was 75% even after 48 hours. The initial color tone of the test section 3 was visually maintained.

(Example 4) 0.01% of α-glucopyranosyl hesperidin was added to a solution containing 0.05% (% by weight) of crocin (carotenoid pigment) which is a gardenia pigment.
%, 0.05%, and 0.1% (weight%) were added and put in a test tube to make test sections 1, 2, and 3, respectively. These were left to stand at 4 ° C., irradiated from all sides with a 12,000 lux fluorescent lamp, and the absorbance was measured at the maximum absorption of crocin at 442 nm over time. The absorbance at the start of standing was set to 100, and the amount of residual dye remained. Is shown as a percentage. At this time, a crocin solution containing no solubilized hesperidin was used as a control. As a result, the initial color tone of α-glucopyranosyl hesperidin was visually maintained in the test sections 2 and 3.

(Example 5) 0.01% solubilized hesperidin was added to a juice having a formulation shown in Table 2 containing 0.05% (wt%) of red cabbage pigment (containing anthocyanin) in an amount of 0.01%.
Add 05% and 0.1% (wt%) and make juice by the usual manufacturing method, put in a test tube and put in test sections 1, 2, 3 respectively.
And These were allowed to stand still at 4 ° C., irradiated with a 12,000 lux fluorescent lamp from all sides, and the maximum absorption of anthocyan of 538 nm was measured with time. At this time, juice containing no solubilized hesperidin was used as a control. The control group gradually discolored (decrease in absorbance) with the lapse of time, and the residual amount of the dye became 50% after 24 hours. On the other hand, the dyes in the test plots were stabilized, and the residual amount of the dye after 24 hours was 75% or more in all the test plots. It was judged that these had sufficient commercial value because the initial color tone was visually maintained. Further, in the test section 3, the color tone was stable and the residual amount of the dye did not decrease even after 24 hours. Further, with the above addition amount, there was no change in the taste and color tone of the juice due to the solubilized hesperidin.

[Table 2]

Example 6 Phycocyanin 0.1%
(% By weight) 0.01%, 0.05%, and 0.1% of solubilized hesperidin in ice cream having the composition shown in Table 3
(% By weight), ice cream was prepared by the usual production method, and put into transparent plastic containers (without lid) to give test sections 1, 2, and 3, respectively. These were allowed to stand at −20 ° C., irradiated from all sides with a 12,000 lux fluorescent lamp, dissolved over time, and the maximum absorption of phycocyanin at 620 nm was measured. At this time, an ice cream containing no solubilized hesperidin was used as a control. In the control group, the color gradually faded (absorption decreased) with the lapse of time, and after 48 hours, the residual amount of the dye became 35%. In comparison, the dye in the test section was stabilized, and the residual amount of the dye after 48 hours was in the test section 1,
It was 70%, 75%, and 85% for 2 and 3, respectively. In particular, in the test plots 2 and 3, it was visually judged that the initial color tone was maintained and the product had a sufficient commercial value. Further, at the above addition amounts, the taste and color tone of the ice cream due to the solubilized hesperidin did not change.

[Table 3]

Example 7 The same test as in Example 1 was conducted on the solubilized naringin. As a result, similar results were obtained with solubilized hesperidin.

Example 8 The same test as in Example 4 was conducted on α-glucopyranosyl naringin. As a result, similar results were obtained with α-glucopyranosyl hesperidin.

[0033]

[Effect] When solubilized hesperidin is added to an object containing colorants and vitamins, it has an effect of absorbing ultraviolet rays such as sunlight and fluorescent lamps and preventing the colorants and vitamins from being destroyed. Further, since the dissolved solubilized hesperidin is colorless and transparent, the color tone of the object does not change by adding it. Therefore, a large amount of solubilized hesperidin can be added to the target.

[0034]

[Brief description of drawings]

FIG. 1 shows absorption spectra of solubilized hesperidin and solubilized rutin.

FIG. 2 Comparison of HPLC patterns of hesperidin glycosides before and after β-amylase treatment Left diagram; before β-amylase treatment Right diagram; after β-amylase treatment A; Hesperidin triglyceride B; Hesperidin diglyceride C; Hesperidin monoglyceride D; Hesperidin

Claims (6)

[Claims] 1. A method for stabilizing foods and the like containing colorants and / or vitamins, characterized in that solubilized hesperidin is added. 2. The method for stabilizing foods and the like containing a coloring agent and / or vitamin according to claim 1, wherein the solubilized hesperidin is α-glucopyranosyl hesperidin. 3. A strain A2-5a whose solubilized hesperidin is of the genus Bacillus (Deposit F of the Institute of Biotechnology, Institute of Industrial Science and Technology)
CG harvested from a culture of ERM P-13864)
The method for stabilizing foods or the like containing a coloring agent and / or vitamin according to claim 1, characterized in that it is produced using Tase. 4. A method for stabilizing foods and the like containing colorants and / or vitamins, characterized in that solubilized naringin is added. 5. The method for stabilizing foods and the like containing a coloring agent and / or vitamin according to claim 1, wherein the solubilized naringin is α-glucopyranosyl naringin. 6. The solubilized naringin strain Bacillus strain A
2-5a (Institute of Biotechnology, Institute of Biotechnology, Fungus Deposited FE
CGT harvested from cultures of RM P-13864)
The method for stabilizing foods and the like containing a coloring agent and / or vitamin according to claim 1, characterized in that it is produced using ase.