JPS6016933A - Antimutagenic agent - Google Patents

Abstract

PURPOSE:To provide the titled agent capable of lowering or eliminating the mutagenicity of food, table luxuries, etc., and composed of an embryo preparation such as malt, soybean, rice bran, and their extracted fractions, etc. CONSTITUTION:An antimutagenic agent is obtained by using an embryo preparation selected from the group of malt, soybean, rice bran, and their extracted fractions, as a component. The mutagens proved to be existent in foods and table luxuries are e.g. benzo[a]pyrene, aflatoxin B1, 2-aminofluorene, benz[a] anthracene, etc. In the usual foods and drinks, the level of mutagenicity is high in coffee. The above embryo preparation inactivates the mutagenicity in black tea and straight bourbon as well as coffee. The necessary amount of the preparation is 0.25-25mg of the malt powder, soybean flour or rice bran, or 0.05-0.5mg of the dried powder of said extracted fraction based on the normal concentration of coffee (5-20mg of coffee powder per 1 liter).

Description

DETAILED DESCRIPTION OF THE INVENTION Technical Field The present invention relates to an antimutagenic agent that reduces or eliminates the mutagenicity of foods, luxury goods, and the like. More specifically, the present invention relates to an antimutagenic agent comprising germ preparations such as malt, soybean, rice bran, and their respective extract fractions.

■) Background Art Currently, most cancers in humans are thought to be caused by carcinogenic factors in the living environment. It is estimated that only about 2% of human cancers are caused by genetic factors alone (Higginson).

J., & Muir, C.S.: J., Natl., Ca.
ncer In5t,, Vol.

63.1291. : 79),.

(1) The foods and drinks we consume on a daily basis are considered the most important cause of cancer, which accounts for about 4 to 15 deaths in humans, and there is too much epidemiological data to support this view. In recent years, the Salmonella test (also known as the Ames method), which uses histidine requirement as an indicator, has been established as a means of predicting the carcinogenicity of environmental factors such as foods, drinks, and pharmaceuticals.
Ames, B.N., et al.: Mutatio
n ILes,, vol 31, 347t '75).

Many research institutions have found a high correlation of 80-90% between the mutagenicity and carcinogenicity of the compounds investigated in this study (McCann, J. et al.
: Prnc, Natl, Acad, Sci. (U, S,
A.), VoI.

72.5135. '75).

Therefore, eliminating mutagenicity is expected to have a considerable effect on protecting humans from the risk of carcinogenesis.7 Not only that, mutations occur as the result of damage to chromosomes or DNA, and even if they do not cause carcinogenesis. The effects of mutagens on human health cannot be underestimated, even if they do not reach the same level as human health.

(2) This is why food and drink products that are not mutagenic are desired.

Today, mutagens that are clearly found in foods, drinks, and luxury goods include benzo[a]pyrene and aflatoxin B.
These include 1.2-aminofluorene, benz[a]anthracene, chrysene, dimethylnitrosamine, β-naphthylamine, nitrosopyrrolidine, methylglyoxal, and many other substances that have been proven to be carcinogenic. but,
No matter how much mutagenicity is recognized, it is a hot topic to take legal prohibition measures in the case of products that have been closely connected to human life, such as food, drinks, and luxury goods. It is also difficult to live without it.

Many of the luxury foods for men and women have higher mutagenicity than everyday foods and drinks (see Table 1).

(3) Table 1 Mutation activity of luxury goods Qin Yu Favorite goods Number of reversion mutation colonies 1 cigarette 5500 (TA98.+) 1 cup of instant coffee (150+++/) 45DOO (
TA, 100. -) 1 cup of regular coffee (15 marks d
) 195D00 (TAloo, -) 1 cup of black tea (150
m7! ') 38,000 (TAloo, -) 1 cup of green tea (
150m1) 21,000 (TAloo, -) Sencha 1
Cup (1,50m1), 17,000 (TAloo, -
) Salmonella typhimurium T
A100 strain or TA98 strain was used.

*◆ Indicates the presence or absence of rat liver homogenate (89 mix).

(-1-1: 89 mixes added, Sun: 89 mixes not added.

In fact, it is often reported that there is a correlation between tobacco and oropharyngeal cancer, pharyngeal cancer, esophageal cancer, stomach cancer, and lung cancer, coffee and pancreatic cancer, bladder cancer, and alcoholic beverages and esophageal cancer and liver cancer.

The present invention aims to improve the safety of foods or luxury goods by eliminating or dissociating them from mutagenicity (4), thereby restoring the reliability of the goods. It is something.

It is said that coffee was introduced as a daily drink in Arabia in the 1400s. Since then, as human society has become more highly civilized, the demand for it as a luxury item has increased more and more. Japan has also recently recorded rapid growth in coffee consumption (approximately six times in the ten years since 1969).

However, in recent years, concerns have begun to be raised about coffee's negative effects on health in various ways, and it is believed to be a major reason why coffee consumption in the United States slumped last year. That is,
The relationship between caffeine and cancer, including its cytotoxicity, and in particular the correlation between coffee drinking and pancreatic cancer pointed out by Dr. MacMain, caused a sensation mainly in Europe and the United States.
Mahon.

B, et at, :N, Engl:J, Med, V
ol, 304 + 630° 1981). Other than pancreatic cancer, the other organs where increased carcinogenicity is seen due to coffee are the lower urinary tract, including the bladder (Simon, D.
, et al. :J, Natl(5) Cancer In5t,, Vol, 54,587.1
975).

Furthermore, there are reports that it increases the incidence of prostate cancer, leukemia, and ovarian cancer, so more detailed investigation is awaited in the future.

Regarding the mutagenicity mentioned above, coffee has a high level among everyday foods and drinks (see Table 1).

Nowadays, coffee is consumed in large quantities as a daily drink, and there is an urgent need to eliminate the various points that are considered harmful to health. Research in this field has produced many results to date. For example, peroxidase against the carcinogen Trp-P-1, N-methyl-N/-nitro-
Thiol compounds for N-nitrosoguanidine, 7.
It has been reported that selenium compounds for 12-dimethylbenzanthracene have mutagenic inactivating effects on carcinogens (Martin, 8.E, et al.
, :MutationRes,, Vol. 82, 41
．． 1981). However, many inactivating agents are themselves harmful to living organisms or are unsuitable for regular consumption due to palatability, making it difficult to put them into practical use.

(6) By using substances that are harmless to humans and do not impair the original characteristics of coffee such as taste and aroma,
Eliminating stasis and mutagenicity is absolutely necessary to protect human health.

(0 Disclosure of the Invention In order to achieve the above-mentioned object, the present inventors have made extensive research efforts and hereby present the extracts of malt, soybean, rice bran, and their respective extract fractions (hereinafter collectively referred to as 'germ preparations'). We have discovered the fact that this ingredient has remarkable efficacy as an inactivator for mutagenicity in coffee.The above ingredients can be used to produce non-mutagenic foods or luxury goods, or to reduce the mutagenicity of foods or luxury goods. The use of the above-mentioned ingredients to eliminate coffee is completely new.The amount of germ preparation required to suppress the mutagenicity of coffee is approximately the same as the concentration of coffee normally consumed by humans (5 to 2 o Da/TLl as coffee powder). ) K for malt flour, soybean flour, rice bran: o, 25-2.5~, malt, soybean, and rice bran extract fraction dry powder: 0.05-0.5■
With this addition amount, the following effect (7) can be obtained. However, when drinking coffee as hot coffee, it is inevitable that the activity will be reduced to some extent, so it would be best to use an amount 2 to 6 times the amount added above. This has led to the spread of Salmonella typhimurium (Salmonella Ia) in coffee.
typhjmurium) TA 100 strain is eliminated by approximately 50% to 100%.

For coffee, coffee beans or coffee powder can be mixed with germ preparations (regular type), or germ preparations can be added to coffee liquid extracted from roasted coffee beans (liquid coffee type, for example, canned coffee). Alternatively, you can homogeneously mix the germ preparation into powdered coffee by spray-drying or freeze-drying, or spray the same into the coffee powder as a liquid (
It can be produced by fluidized bed granulation (instant coffee type). Furthermore, when drinking, it may be mixed into sugar, coffee cream, powdered cream, etc. in the form of powder or solution. (8) Alternatively, the desired germ preparation can be immersed and attached to a filter paper or cloth for dispersing regular coffee infusion.

Coffee whose mutagenicity has been reduced or completely inactivated by water droplets has excellent palatability and is therefore considered more desirable for the purpose of reducing risks to human health.

In addition to coffee, the germ preparation also inactivates the mutagenic effects of tea and straight bourbon. Regarding tea,
After drying by fire, mutagenicity can be reduced by mixing the germ preparation into powder or by attaching it to tea leaves using a fluidized bed granulation method. It may still be mixed in powder or solution form when drinking. For straight bourbon, whether the germ preparation is included in the product itself or
Its mutagenicity can be inactivated by adding it in powder or solution form at the time of drinking. The present invention is based on the above novel findings.

(9) Example 0■) Method for measuring mutagenicity and mutagenicity suppressing effect (1) Method: Frein incubation method (cedar wood, nagaya;
Chemical Mutagiens, Volume 6, Page 41, 1981
)by.

(11) Bacterial strain used: histidine-requiring Salmonella
Typhimurium (Salmonella typhi)
murium) Ti2O3 strain (hereinafter abbreviated as "S, TA100 strain").

011) Preparation of sample a) For instant coffee: Dissolve instant coffee (powder) in distilled water. On the other hand, a predetermined amount of the germ preparation is also suspended or dissolved in distilled water, and 50 μl of each solution is mixed.

Mouth) For regular coffee: 2507Wf per 20g of roasted coffee beans! ! Extract with hot distilled water and filter the extract with coffee filter paper (Kalita Seiya 12). Freeze-dry the r solution, measure its dry weight, and then dissolve it in distilled water. On the other hand, a predetermined amount of the embryo preparation is also suspended or dissolved in distilled water, and (1o) 50 μl of both solutions are mixed.

c) For black tea: Extract 5 to 7 g of commercially available black tea with 200 md of boiled distilled water. After extraction, the leaves are removed, the extract is freeze-dried, and distilled water is added to the dried product to dissolve it to a predetermined concentration. On the other hand, a predetermined amount of the germ preparation is suspended or dissolved in distilled water, and both solutions are mixed at a ratio of 1=4 to make a total of 100 μl.

2) In the case of bourbon: Commercially available bourbon (also known as straight bourbon or bourbon whiskey) is concentrated to dryness under reduced pressure at 40° C. using a rotary evaporator. After redissolving in DMSO, suspend or dissolve a predetermined amount of the germ preparation in distilled water.1. , both solutions were mixed in equal amounts to make a total of 1001 tll.

e) Germ preparation: Among the germ preparations, rice bran was prepared as is, malt and soybean were ground and then distilled water was added to a predetermined amount.

Each extracted fraction dried powder was obtained by extracting 1 g of crushed malted soybeans and rice bran with 10 ml of distilled water at 4°C for 160 minutes.
After centrifuging the extract (11) (1,000xy), the supernatant was filtered through a Millipore filter (pore size: 0.45 μm), and the filtrate was freeze-dried and powdered. A predetermined amount of this was dissolved in distilled water and used for experiments.

To 100 μl of each sample obtained according to ivl Mutagenicity Measurement Pre-book, mark ~), add 100 μl of S, TA10 to 500 μl of 0.1 M sodium phosphate buffer (pH 7,4).
Add 0 strain culture solution. This mixture was heated to 67°C.
After shaking for 20 minutes, mix with 2rnl of dissolved soft agar and spread on a 0.1% glucose agar plate. In addition, 0.1 micromol/2 ml of soft agar/plate of histidine, which is necessary to divide the bacteria several times on the plate, is added to the soft agar.

After standing at 37°C for 48 hours, the number of colonies on the plate is counted as a revertant strain. In addition, the mutagenicity suppression rate is calculated using the following formula.

(12) ■) Results (1) Mutagenicity suppressing effect of germ preparation on coffee.

(a) Regular coffee Coffee bean extract prepared as described above Dissolve 15 μl of the extract powder in distilled water to make 50 μl.

On the other hand, a predetermined amount of the germ preparation is suspended or dissolved in distilled water to a total volume of 50 μl, and the two are mixed and allowed to stand at room temperature for 20 minutes.

This was supplemented with 0.1 M sodium phosphate buffer (pI 47.
4) 0.5m/ and the culture solution of the above S and TA100 strains 0
．． The mutagenicity was measured using the pressure method described in (C)I) (1) above, and the results shown in FIG. 1 were obtained.

The mutagenicity of regular coffee can be determined by adding malt flour, soybean flour, or rice bran to 5% (W/W) of the extracted coffee powder, or by adding 1% (W/W) malt or rice bran to the extracted coffee powder.
Most of it can be inactivated by adding dry powder extracted from soybeans and rice bran. For complete inactivation of mutagenicity, the amount added to coffee powder must be 20% as malt flour, soybean flour, and rice bran, and 4% as (IB) malt, soybean, and rice bran extract dry powder. be.

(b) Instant coffee Using commercially available instant coffee powder, the antimutagenic activity of the germ agent was examined in the same manner as for regular coffee. The results are shown in Figure 2.

In the case of instant coffee, mutagenicity was significantly reduced or completely eliminated by the germ preparation, as was the case with regular coffee mentioned above. In other words, Instagram 2/
I. For coffee powder, add 5 parts of malt powder, soybean flour, and rice bran.
% (W/W), 1. y, c W/W ), most of its mutagenicity can be inactivated. Mutagenicity can be completely eliminated if the amount added to the coffee powder is 20% as malt flour or soybean flour, and 4% as malt, soybean, or extracted fraction dry powder. In addition, in the case of rice bran,
They are 30% and 6%, respectively.

Regular coffee, instant coffee and (14) were once inactivated by the addition of a 39 mix (a combination of 9oOo,y supernatant of rat liver homogenate and reduced nicotinamide adenine dinucleotide phosphate (NADPH) producing thread). The mutagenicity that had developed was never to appear again.

(11) Inhibitory effect of germ preparation on black tea mutagenicity.

The black tea prepared by the above-mentioned method is as shown in Figure 3.
It has mutagenicity against Salmonella enterica (S, TAloo) at a frequency of approximately 200/108 viable bacteria per ml. On the other hand, the mutagenicity of malt, soybean, and rice bran was significantly reduced by adding 20% or 4% dry powder of malt, soybean, and rice bran extracts, respectively. .

Qii) Inhibitory effect of germ preparations on the mutagenicity of straight bourbon.

Straight bourbon is made of L sea urchin, S, as shown in Figure 4.
, has mutagenicity (approximately 800 revertant colonies/108 viable bacteria) (1.5) against TAloo. On the other hand, when malt flour, soybean flour, and rice bran are added at a concentration of 20% (W/W), or each extract fraction is added at a concentration of 4% (W/W) as dry powder, their mutagenicity is reduced. Approximately 40-85% was inactivated.

(G) Beyond industrial applicability, coffee and other beverages that have had their mutagen reduced or eliminated using germ preparations have lost their original taste and aroma. Furthermore, since the germ preparation itself is rich in vitamins, it can be commercialized as an additive for producing more desirable types of beverages and foods for human health.

[Brief explanation of the drawing]

Figure 1 is a graph showing the inhibitory effect of germ preparations on the mutagenicity of Salmonella typhimurium TA100 strain in regular coffee; a) Figure is malt flour and its extracted fraction dry powder; b) Figure Figure C) shows dried powder of soybean flour and its extracted fraction, and Figure C) shows dried powder of rice bran and its extracted fraction. (xi) In Figure 1, * indicates additive-free coffee, and M, M, and ■ indicate 2.
0% (W/W') of malt flour, soybean flour, and rice bran are added to coffee. show. Figure 2 is a graph showing the inhibitory effect of germ preparations on the mutagenicity of Salmonella typhimurium TA100 strain in instant coffee. and soybean extract fraction dry powder, C) Figure is a graph about rice bran and rice bran extract fraction dry powder.・, Mu, Ma, △, , ■ in FIG. 2, and the dry taste in the mouth are the same as in FIG. 1. FIG. 3 is a graph showing the reduction of black tea mutagenic germ preparation against Salmonella typhimurium strain TA100. In Figure 3, ・ indicates black tea without the addition of germ preparations, and MU, MA, and ■ indicate 20% malt, soybean flour, and rice bran (W/W).
) △,, Mouth is 4% dry powder extracted from malt, soybean, and rice bran.
(W/W) Indicates added black tea. (17) FIG. 4 is a graph showing the reduction of bourbon mutagenicity against Salmonella typhimurium strain TA100 by germ formulation. In Figure 4, * is bourbon without the addition of germ preparations, and MU, MA, and ■ are bourbon containing malt flour, soybean flour, and rice bran.
0% (W/W), △, Mouth indicates bourbon to which 4% (W/W) of dry powder extracted from malt, soybeans, and rice bran was added. Patent applicant Suntory Ltd. (18) -4-q. Mr. Kazuo Wakasugi, Commissioner of the Japan Patent Office, 1 cup. Indication of the case Patent Application No. 124244 of 1982 2, Name of the invention Anti-mutagenic agent 6, Person making the amendment Relationship to the case Note: Name of applicant's address (190) Zantry Co., Ltd. 4, Attorney's specification [Detailed Description of the Invention] Column 6, the content of the amendment is corrected as follows: Page Line Before correction After correction 6 17 1981), 1981, etc.), 1 row 11
Previous (C) ■) Previous (D) I) 17 1 Ma, ^ Mu,
Ma or above

Claims (1)

[Claims] An antimutagenic agent comprising an germ preparation selected from the group consisting of malt, soybean, rice bran, and their respective extract fractions.