阅读说明：本技术 一种甜菊糖甙rb的提纯方法 (Purification method of stevioside RB ) 是由 郑越 于 2019-09-30 设计创作，主要内容包括：本发明公开了一种甜菊糖甙RB的提纯方法,具体涉及植物提纯技术领域,包括以下步骤：步骤一,取甜菊糖甙母液,将母液倒入至坩埚内,并向坩埚内添加氧化钙粉末后搅拌均匀,静置1-3小时,然后再向坩埚内添加2mol·L<Sup>-1</Sup>的FeSO4溶液,搅拌均匀后,将其加热并浓缩至原质量的40%,获取A相溶液；步骤二,将蒸馏水浸泡好的阴离子交换树脂和阳离子交换树脂分别装入离子交换柱中,并按先阴离子后阳离子的顺序连接好装置。本发明提供的提纯方法,能够制得RB含量45%以上的甜菊糖甙,提供了以RB为主的甜菊糖甙产品,满足了消费者的不同需求,能够适应多样化的市场需求,备要求低,可操作性强,具有良好的社会推广应用。(The invention discloses a method for purifying stevioside RB, which particularly relates to the technical field of plant purification and comprises the following steps: step one, taking stevioside mother liquor, pouring the mother liquor into a crucible, adding calcium oxide powder into the crucible, uniformly stirring, standing for 1-3 hours, and then adding 2 mol. L into the crucible ‑1 Uniformly stirring the FeSO4 solution, heating and concentrating the solution to 40% of the original mass to obtain an A phase solution; and step two, respectively filling the anion exchange resin and the cation exchange resin soaked in the distilled water into an ion exchange column, and connecting the devices according to the sequence of anion and cation. The purification method provided by the invention can prepare the stevioside with the RB content of more than 45 percent, provides the stevioside product mainly containing RB, meets different requirements of consumers, can adapt to diversified market requirements, and is ready for useThe method has the advantages of low requirement, strong operability and good social popularization and application.)

1. A method for purifying stevioside RB is characterized by comprising the following steps:

step one, taking stevioside mother liquor, pouring the mother liquor into a crucible, adding calcium oxide powder into the crucible, uniformly stirring, standing for 1-3 hours, and then adding 2 mol. L into the crucible^-1Uniformly stirring the FeSO4 solution, heating and concentrating the solution to 40% of the original mass to obtain an A phase solution;

step two, respectively filling anion exchange resin and cation exchange resin soaked by distilled water into an ion exchange column, connecting a device according to the sequence of anion and cation, then adding the phase A solution into the anion exchange column, washing by using ethanol with the volume fraction of 95%, collecting the ethanol solution into a crucible, and heating the crucible to concentrate the ethanol solution to obtain an extract;

step three, recrystallizing the extract by using a methanol solution with volume fraction of 5% to obtain a stevioside crystal, adding the stevioside crystal into a wide-mouth beaker, pouring deionized water with the mass ratio of 5:1 to the crystal, heating the beaker to enable the deionized water to be heated to 60-80 ℃ and the stevioside crystal to be melted, and then preserving heat for 3-5 hours to obtain a C-phase solution;

pouring the C phase solution into a container, heating to 110 ℃ in a high-pressure environment, boiling the solution to evaporate water in the solution, stopping heating after the solution is concentrated to 50% of the initial mass, and pouring the solution into a cylindrical container after the solution is cooled to the normal temperature;

inserting a large-aperture adsorption resin column with the diameter smaller than the inner diameter of the cylindrical container into the cylindrical container, immersing the whole adsorption resin column in the solution, performing differential adsorption on the polarities of different components in the solution by using the adsorption resin column, taking out the adsorption resin column after soaking for 10-15 hours, resolving the stevioside adsorbed on the resin column by using 1200L of 75% ethanol, collecting eluent, and measuring the RB content in the eluent by using a liquid phase method.

2. The method of claim 1, wherein the purification of RB comprises: wherein, 1200L of 75% ethanol is used for resolving the stevioside adsorbed on the resin column, then the eluent is collected and concentrated at the temperature of 60 ℃, the solid content after concentration is controlled at 50%, the obtained solid and liquid are respectively dried to obtain the stevioside, and the RB detection is carried out by a liquid phase method, and the RB content of the stevioside is detected to be 47.64%.

3. The method of claim 1, wherein the purification of RB comprises: wherein, 1200L of 75% ethanol is used for resolving the stevioside adsorbed on the resin column, then the eluent is collected and concentrated at 70 ℃, the solid content after concentration is controlled at 50%, the obtained solid and liquid are respectively dried to obtain the stevioside, and the RB detection is carried out by a liquid phase method, and the RB content of the stevioside is detected to be 49.37%.

4. The method of claim 1, wherein the purification of RB comprises: wherein, 1200L of 75% ethanol is used for resolving the stevioside adsorbed on the resin column, then the eluent is collected and concentrated under the condition of 80 ℃, the solid content after concentration is controlled at 50%, the obtained solid and liquid are respectively dried to obtain the stevioside, and the RB detection is carried out by a liquid phase method, and the RB content of the stevioside is detected to be 53.81%.

5. The method of claim 1, wherein the purification of RB comprises: in the first step, the particle mesh number of the calcium oxide powder is 300-500 meshes.

6. The method of claim 1, wherein the purification of RB comprises: in the fourth step, the high pressure is 1.42 standard atmospheric pressures.

Technical Field

The invention relates to the technical field of plant purification, in particular to a method for purifying stevioside RB.

Background

Stevia rebaudiana (Bertoni) Hemsl, a scientific name, is a perennial herb of the Stevia genus of the Compositae family. The plant height is 1-1.3 m. The root tips are large, 50-60 strips, and the length can reach 25 cm. Upright stem, lignified shoot at the base, tender upper part, dense short hairy hair, purple red or white corolla at the base and white upper part. The thin fruit is linear, slightly flat and brown and has crown hair. The flowering period is 7-9 months, and the fruit period is 9-11 months. Native to alpine grasslands bordering paraguay and brazil. Since 1977, China Beijing, Hebei, Shanxi, Jiangsu, Anhui, Fujian, Hunan, Yunnan and other places have been introduced for cultivation. The species prefers to grow in a warm and humid environment and is sensitive to light. The leaf contains 6-12% of inulin glycoside, and the refined product is white powder, is a natural sweetener with low calorie and high sweetness, and is one of raw materials in food and pharmaceutical industry.

The stevia rebaudiana is widely applied to the fields of food, beverage, wine making, medicine, cosmetics and the like after being refined. In recent years, stevia glycosides, an extract of stevia rebaudiana is more used as a sweetener. The sweetener has very low calorie and heat energy, does not cause adverse reaction to patients with odontopathy and diabetes, becomes an excellent sweetener, and has wide market prospect. Stevioside is a general name of stevia extract, and currently, in the official method of FDA in the united states, the following 9 components are mainly involved in relation to stevioside: stevioside (Stevioside), Rebaudioside A (Rebaudioside A), Rubusoside F (Rebaudioside F), Rebaudioside D (Rebaudioside D), Steviolbioside (Steviolbioside), Rebaudioside B (Rebaudioside B), and Stevioside RB is one of the components Rebaudioside B. These different components all have different tastes and can be confronted with different consumer groups, such as: consumers in the Mega area prefer RA, while consumers in Japan and Korea prefer STV.

At present, stevioside products on the market are mainly RA and STV, and products mainly comprising RB are not available, so that the extraction method of stevioside is mainly concentrated on the purification and refining of RA and STV, and a better purification method of RB is not available. Therefore, it is a market demand to provide a method for purifying stevioside RB to meet the diversified needs of consumers.

Disclosure of Invention

In order to overcome the above defects of the prior art, embodiments of the present invention provide a method for purifying rebaudioside b, wherein after a stevioside crystal is mixed with deionized water, evaporating water under high pressure, concentrating the solution, introducing an adsorbent resin column into the container, adsorbing different components of the solution with different polarities by the adsorbent resin column to separate RB, and then, the recrystallization of the eluent is completed, the temperature is accurately controlled, the stevioside with the RB content is obtained, the energy consumption is low in the whole process, no waste water is generated, the environment is more protected, the stevioside with the RB content of more than 45% can be prepared, the stevioside product mainly containing RB is provided, different requirements of consumers are met, the method can adapt to diversified market demands, the preparation requirement is low, the operability is high, and the method has good social popularization and application.

In order to achieve the purpose, the invention provides the following technical scheme: a method for purifying stevioside RB comprises the following steps:

step one, taking stevioside mother liquor, pouring the mother liquor into a crucible, adding calcium oxide powder into the crucible, uniformly stirring, standing for 1-3 hours, and then adding 2 mol. L into the crucible^-1Uniformly stirring the FeSO4 solution, heating and concentrating the solution to 40% of the original mass to obtain an A phase solution;

step two, respectively filling anion exchange resin and cation exchange resin soaked by distilled water into an ion exchange column, connecting a device according to the sequence of anion and cation, then adding the phase A solution into the anion exchange column, washing by using ethanol with the volume fraction of 95%, collecting the ethanol solution into a crucible, and heating the crucible to concentrate the ethanol solution to obtain an extract;

step three, recrystallizing the extract by using a methanol solution with volume fraction of 5% to obtain a stevioside crystal, adding the stevioside crystal into a wide-mouth beaker, pouring deionized water with the mass ratio of 5:1 to the crystal, heating the beaker to enable the deionized water to be heated to 60-80 ℃ and the stevioside crystal to be melted, and then preserving heat for 3-5 hours to obtain a C-phase solution;

pouring the C phase solution into a container, heating to 110 ℃ in a high-pressure environment, boiling the solution to evaporate water in the solution, stopping heating after the solution is concentrated to 50% of the initial mass, and pouring the solution into a cylindrical container after the solution is cooled to the normal temperature;

inserting a large-aperture adsorption resin column with the diameter smaller than the inner diameter of the cylindrical container into the cylindrical container, immersing the whole adsorption resin column in the solution, performing differential adsorption on the polarities of different components in the solution by using the adsorption resin column, taking out the adsorption resin column after soaking for 10-15 hours, resolving the stevioside adsorbed on the resin column by using 1200L of 75% ethanol, collecting eluent, and measuring the RB content in the eluent by using a liquid phase method.

In a preferred embodiment, 1200L of 75% ethanol is used for resolving the stevioside adsorbed on the resin column, then the eluent is collected and concentrated at 60 ℃, the solid content after concentration is controlled to be 50%, the obtained solid and liquid are respectively dried to obtain the stevioside, and the RB detection is carried out by a liquid phase method, and the RB content of the stevioside is detected to be 47.64%.

In a preferred embodiment, 1200L of 75% ethanol is used for resolving the stevioside adsorbed on the resin column, then the eluent is collected and concentrated at 70 ℃, the solid content after concentration is controlled to be 50%, the obtained solid and liquid are respectively dried to obtain the stevioside, and the RB detection is carried out by a liquid phase method, and the RB content of the stevioside is detected to be 49.37%.

In a preferred embodiment, 1200L of 75% ethanol is used for resolving the stevioside adsorbed on the resin column, then the eluent is collected and concentrated at 80 ℃, the solid content after concentration is controlled to be 50%, the obtained solid and liquid are respectively dried to obtain the stevioside, and the RB detection is carried out by a liquid phase method, and the RB content of the stevioside is detected to be 53.81%.

In a preferred embodiment, in the first step, the particle size of the calcium oxide powder is 300-500 mesh.

In a preferred embodiment, in step four, the high pressure is specifically 1.42 atm.

The invention has the technical effects and advantages that:

1. the purification method provided by the invention can prepare the stevioside with the RB content of more than 45 percent, provides the stevioside product mainly containing RB, meets different requirements of consumers, can adapt to diversified market requirements, has low preparation requirement and strong operability, and has good social popularization and application;

2. after the stevioside crystal is mixed with deionized water, evaporation of water is completed under a high-pressure environment, the solution is concentrated, a cylindrical container is used, an adsorption resin column is inserted into the container, the adsorption resin column is used for distinguishing and adsorbing polarities of different components in the solution, RB is separated out, then recrystallization of eluent is completed, the temperature is accurately controlled, the stevioside with the RB content is obtained, energy consumption in the whole process is low, no waste water is generated, and the method is more environment-friendly.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.