JPH0375140B2 - - Google Patents

Description

[Detailed description of the invention]

This invention focuses on rebaudioside A relative to stevioside among sweet components contained in stevia.
This invention relates to a stevia natural sweetener with a high content of rebaudioside C and a relatively low content of rebaudioside C. Stevia is a perennial plant of the Asteraceae family that is native to Paraguay in South America, and its scientific name is Stevia Rebaudiana.
Bertoni). Stevia contains a sweet component that is over 300 times sweeter than sugar, so it is cultivated to extract this sweet component and use it as a natural sweetener. The sweet component of stevia is stevioside (C ₃₈ H ₆₀ O ₁₈ , molecular weight 804), which has the following structural formula: Rebaudioside A having the following structural formula (C ₄₄ H ₇₀ O ₂₃ , molecular weight 966) and rebaudioside C (C ₄₄ H ₇₀ O ₂₂ , molecular weight 950, also called dulcoside B) having the following structural formula: In addition, rebaudioside D, E, zurcoside A, etc. are known. In commonly cultivated stevia varieties, stevioside is the main component among the above sweetening components, with the content of rebaudioside A being about three-tenths of that of stevioside, and the content of rebaudioside C being slightly lower. (4th
(see figure), depending on the variety, rebaudioside A
and those containing no C, and those containing rebaudioside C as a main component. Stevioside is used in the food industry as a natural sweetener because it is 300 times sweeter than sugar. Its sweet taste is relatively similar to sugar, but it has the disadvantage of leaving an unpleasant aftertaste such as bitterness. In addition, rebaudioside C is 40% of sugar.
Not only is it 60 times as sweet, but its sweetness also has a strong astringent taste. Therefore, containing large amounts of stevioside and rebaudioside C,
It is not desirable as a sweetener. On the other hand, rebaudioside A is similar to stevioside.
It has a sweetness level of 1.3 to 1.5 times, and its sweetness is similar to sugar, mild and does not leave any unpleasant taste. Therefore, this inventor discovered that rebaudioside A
The aim was to create a sweetener with a high content of Stevioside and Rebaudioside C and a low content of Stevioside and Rebaudioside C. It is already known that stevioside in a stevia extract can be removed to some extent by methods such as crystallization. By removing stevioside in this manner, it is possible to obtain a sweetener having a low stevioside content and a relatively high rebaudioside A content as a mother liquor. However, even if attempts were made to remove rebaudioside C by crystallization using various solvents, it was not possible to selectively remove rebaudioside C. Not only that, but when stevioside was removed as described above, the content of rebaudioside C in the mother liquor was relatively increased, and the quality of the sweetness was even worse. Moreover, impurities with poor taste other than rebaudioside C were also concentrated in the mother liquor, which adversely affected the quality of sweetness. In order to improve the above drawbacks, this inventor
First, we attempted to improve the variety of stevia. And 1.5 rebaudioside A to stevioside.
We succeeded in growing a new variety that contains ~2.55 times as much rebaudioside C and 0.3 to 0.4 times as much rebaudioside C, and by extracting this, we have developed a new variety that contains rebaudioside A 1.5 to 2.55 times as much as stevioside. This invention was completed by confirming that a sweetener containing 0.3 to 0.4 times more Side C could be obtained. That is, this invention provides rebaudioside A that is 1.5 to 2.55 times as much as stevioside and 0.3 to 0.4 times as much rebaudioside A as stevioside.
Stevia containing twice the rebaudioside C
It is obtained by extracting the plant body or dried leaves of Rebaudiana Bertoni with water or a water-containing solvent, and separating and collecting sweet components from the resulting extract.
It is a sweetener containing 1.5 to 2.55 times more rebaudioside A and 0.3 to 0.4 times more rebaudioside C than stevioside. The breeding process of the new Stevia variety used in this invention is as follows, but is not limited to this breeding process.
Any breeding or cultivation method may be used as long as Stevia containing 1.5 to 2.55 rebaudioside A and 0.3 to 0.4 rebaudioside C can be obtained. From October to December 1974, Rebaudioside A
Stevia native variety S, which contains 6/10 of stevioside, was artificially hybridized at Morita Chemical Industries Co., Ltd.'s Niimi Plant in Ashami, Niimi City, Okayama Prefecture, and the resulting seeds were harvested in early March 1980. In early May of the same year, the seeds were sown in a greenhouse for growing seedlings in the same place, and in early May of the same year, the sprouted and grown seedlings were transplanted to the field.In early August of the same year, the seedlings with good growth, branching, and large leaves were selected and sweetened. Investigate the ingredient content and select seedlings containing 1:1 or more of rebaudioside A to stevioside,
I set it as SF ₁ . SF ₁ was propagated by cuttings, artificially crossed in a plastic greenhouse from October to December of the same year, and the resulting seeds were used as Showa
The seeds were sown in a greenhouse for seedlings in February 1956, transplanted to the field in late April of the same year, and the sweet component content was investigated in early August of the same year. The seedlings were selected and named the new variety SF ₂ . When SF ₂ was propagated by cuttings, overwintered, and the overwintered plants were propagated by cuttings and division, and the sweet component content of 100 plants of each was investigated, the ratio of rebaudioside A to stevioside was 1:1.5 to 2.55. , rebaudioside C was found to be 1:0.3-0.4. Furthermore, it is also possible to use stevia and its dried leaves obtained by raising seedlings and cultivating a portion of SF ₂ using a propagation method using biotechnology such as tissue culture, as well as its dried leaves, as raw materials for the present invention. . To obtain the sweetener of the present invention, first, the above-mentioned new variety of Stevia is extracted with water or a water-containing solvent. Next, the extract is concentrated as it is, or if necessary, ionic impurities are removed using a cation exchange resin and an anion exchange resin, and then adsorbed on an adsorption resin, eluted with a hydrophilic solvent, and the eluate is concentrated. others,
Conventional purification means such as decolorization can be applied as appropriate.
Other sweeteners, diluents, etc. can be added to the resulting sweetener. It is desirable that the content of rebaudioside A in the sweetener of this invention be as high as possible, and there is no particular upper limit, but in practice, the content of rebaudioside A in the sweetener obtained by extraction and purification from stevia is generally 1:1 to stevioside. Up to about 7. In addition, it is desirable that the content of rebaudioside C be as low as possible, and there is no particular lower limit, but in practice, the content of rebaudioside C obtained by extraction and purification from stevia is approximately 1:0.1 to stevioside. , up to about 1:0.1 to stevioside,
Recrystallization, adsorption resin treatment, and oxygen treatment can further increase the ratio of rebaudioside A to stevioside, and recrystallization can further remove impurities. Next, this invention will be explained by examples, and the effects of this invention will be clarified by comparative examples and test examples. The present inventor is prepared to distribute seeds of the above-mentioned new Stevia variety SF ₂ to those who wish to cultivate them for testing and research purposes. Examples 1 to 3 Dried leaves A (containing 3.6% stevioside, 5.6% rebaudioside A, 1.1% rebaudioside C, Example 1) and dried leaves B (2.9% stevioside) of the new Stevia variety SF ₂ , rebaudioside A5.8%,
Contains 0.9% rebaudioside C, Example 2) and dried leaf C (contains 2.4% stevioside, 6.0% rebaudioside A, 0.9% rebaudioside C,
Extract 20 g of each of Example 3) several times with 10 to 20 times the amount of water until the sweetness is no longer felt, and the extract is soaked in a cation exchange resin (Amberlite IR-120B).
The passing liquid is passed through a column packed with 100 ml of anion exchange resin (Duolite A-4) and a column packed with 100 ml of anion exchange resin (Duolite A-4).
Sweet components are adsorbed through a column packed with 100 ml of XAD-2), washed thoroughly with water, and eluted with 300 ml of methanol. The eluate is concentrated under reduced pressure and dried to obtain a white powder. The powder analysis method and analysis results are shown in the table below. In addition, in the table below, ST is stevioside, RA is rebaudioside A, and RC is rebaudioside C.

[Table] Analysis method High performance liquid chromatography column Lichrosolve NH ₂ Flow rate 2 ml/min Solvent Acetonitrile: Water = 80:20 Measurement wavelength 210 nm Comparative example 1 20 g of dried leaves of a native variety of Stevia from Shibushi-cho, Soo-gun, Kagoshima Prefecture ( Stevioside 7.2%, Rebaudioside A 2.7%, Rebaudioside C 1.2%)
Extraction and post-treatment were carried out in the same manner as in Examples 1 to 3 to obtain a white powder. Comparative Example 2 The same dried leaves as in Comparative Example 1 were extracted and treated with resin in the same manner as in Examples 1 to 3, the eluate was concentrated, and after adding 100 ml of methanol, it was concentrated to 30 ml.
After standing at 4°C for 30 hours, precipitated crystals (mainly stevioside) were removed, and the mother liquor was concentrated and dried under reduced pressure to obtain a pale yellow powder. The analysis results of the powder of Comparative Example 1, the crystals of Comparative Example 2, and the pale yellow powder are shown in the table below.

[Table] Figures 1 to 5 show graphs obtained by analyzing the powders of Examples 1 to 3 and Comparative Examples 1 and 2, respectively. Analysis method Thin layer chromatography scanner Model used Shimadzu chromatography scanner 910 TLC plate Merck silica gel plate 60F254 Developing solution Chloroform: methanol: water = 30:20:4 Color developer 50% sulfuric acid measurement method Reflection zigzag scanning method Measurement wavelength 350nm scan speed 20mm/min Test example 1 0.27% of the powders of Examples 1 to 3 and Comparative example 1
The powder of 2 is made into a 0.32% aqueous solution, and the powder of 2 is made into a 0.4% aqueous solution.
Bitterness was tested using a two-point comparison method by 10 panelists, and the following results were obtained. Note that the numbers indicate the number of people.

[Table] Test Example 2 The powders of Examples 1 to 3 and Comparative Examples 1 and 2 were tested for astringency using the same method as Test Example 1, and the following results were obtained.

【table】

[Table] Test Example 3 The sweetness quality of the powders of Examples 1 to 3 and Comparative Examples 1 and 2 was tested by the same method as Test Example 1, and the following results were obtained.

【table】

[Table] Comparative example 3 Stevioside dried leaves (stevioside 5.36
%, Rebaudioside A 5.85%, Rebaudioside C 2.1%) was extracted and treated with a resin in the same manner as in Example 1-3. After concentrating the eluent to 15 ml and removing as much of the solvent as possible in a vacuum dryer, add 40 ml of methanol, dissolve by heating, and heat at 4°C.
Crystals (mainly stevioside) that precipitated after standing for 40 hours were removed by filtration, and the mother liquor was concentrated and dried under reduced pressure to obtain a pale yellow powder. The analytical values for the crystals and powder are shown in the table below.

[Table] Test Example 4 Aqueous solutions of 0.27% of the powder obtained in Example 2 and 0.33% of the powder of Comparative Example 3 were prepared, and bitterness, astringency, and sweetness were tested using a two-point comparison method by 10 panelists. , I got the following results. 0 people found the powder of Example 2 more bitter than Comparative Example 3 10 people found the powder of Example 2 less bitter than Comparative Example 3 0 people found the powder of Example 2 more astringent than Comparative Example 3 Those who found the powder of Example 2 to be less astringent than Comparative Example 3 10 Those who found the powder of Example 2 to have better sweetness than Comparative Example 3 10 Those who found the opposite of the above 0 From the above results, the sweetener of this invention It can be seen that this sweetener is superior to the sweetener obtained by extracting native Stevia varieties and the sweetener obtained by removing stevioside from the sweetener and having a relatively high content of rebaudioside.

[Brief explanation of drawings]

Figures 1 to 5 show Examples 1 to 5, respectively.
3. The analysis results of the powders obtained in Comparative Examples 1 and 2 are shown. In each figure, ST indicates the peak of stevioside, RA indicates the peak of rebaudioside A, and RC indicates the peak of rebaudioside C.

Claims (1)

[Claims] 1. The plant or dried leaves of Stevia rebaudiana Bertoni containing 1.5 to 2.55 times more rebaudioside A and 0.3 to 0.4 times more rebaudioside C than stevioside are extracted with water or a water-containing solvent. It is characterized by separating and collecting sweet components from the extracted liquid. Rebaudioside A and 0.3 to 0.4
A method for producing a sweetener containing twice as much rebaudioside C.