Sweetening composition and method for producing the same

Abstract

This application discloses a sweetening composition comprising, based on the total weight of the sweetening composition, 70% to 85% mogroside V, 5% to 12% 11-O-mogroside V, 0 to 0.5% siamenoside I, 0 to 0.05% mogroside II and mogroside III, 0 to 0.5% protein, and 0 to 0.5% total sugar. This application also discloses a method for preparing a sweetening composition, comprising an extraction step of extracting monk fruit to obtain an extract; a clarification step of clarifying the extract to obtain a clarified liquid; an ion exchange treatment step of sequentially treating the clarified liquid with a strongly acidic cation exchange resin column and a weakly basic anion exchange resin column to obtain an ion exchange treatment solution; and a multi-stage adsorption separation step of introducing the ion exchange treatment solution into a combination of adsorption resin columns for multi-stage adsorption separation and elution, collecting the eluate, and drying to obtain the sweetening composition, wherein the ion exchange treatment step is performed before the multi-stage adsorption separation step.

Description

Technical Field

[0001] This application relates to the field of extraction technology, and particularly to sweet compositions and methods for preparing the same.

Background Art

[0002] Momordica grosvenori Swingle fruit is the mature fruit of a perennial vine plant of the genus Momordica in the Cucurbitaceae family. Its properties and flavor are sweet and sour and cold. It returns to the meridians of the lungs and large intestine, removes heat, moistens the lungs, smooths the intestines, and has a laxative function. It is a traditional Chinese medicine commonly used in China and is also the first medicinal and edible plant approved by the health department. Momordica grosvenori Swingle fruit is mainly produced in Guilin, Guangxi, and is widely cultivated in Yongfu County, Lingui County, Longsheng County, etc. in Guangxi. In recent years, due to the rapid growth in demand for mogroside sweeteners, the cultivation of Momordica grosvenori Swingle fruit has gradually expanded to cities and counties in Guizhou Province and Hunan Province adjacent to Guilin.

[0003] Mogroside is a general term for sweet cucurbitan-type triterpene saponins found in monk fruit. It is a type of triterpene glucoside, and its aglycone is triterpenol. Among them, mogroside V is the most important sweetening component. It is non-toxic, low in calories, highly sweet, and has good heat stability. It is an important indicator for content control under the Chinese National Food Safety Standard (GB1886.77-2016) for natural sweeteners. Currently, mogroside is widely used in the food, beverage, and dairy industries as a low-calorie natural sweetener to replace sucrose. In the current market, the purity of mogroside V in mogroside products is mainly 40-50%, and products with a purity of 60% or less are rare. These mogroside products contain not only various extremely sweet sweet glycosides, but also small amounts of bitter glycosides such as mogroside II and mogroside III. This is because, during the extraction of fresh monk fruit, bitter components from unripe or under-mature monk fruit are mixed in, negatively affecting the overall taste of the sweet glycoside product. Furthermore, current mogroside products typically still retain the high protein content and certain sugars derived from fresh monk fruit, and the presence of these nutrients imposes certain limitations on the product's shelf life and its use in applications requiring high energy intake. [Overview of the Initiative] [Problems that the invention aims to solve]

[0004] To improve the quality of mogroside products, researchers have also developed several methods for preparing high-purity mogroside V, such as Chinese Patent CN110669095A "Method for Extracting High-Purity Mogroside V" and Chinese Patent CN101402665B "Method for Preparing Mogroside V." While these methods have yielded mogroside V products with a purity of over 99%, all of these processes employ purification techniques centered on reverse-phase chromatography with 0.45 μm particle size C18 fillers. This technology uses medium-to-high pressure preparative liquid chromatography, resulting in significant capital investment and limited mass production capacity. Furthermore, the process uses toxic solvents such as acetonitrile, which is detrimental to food safety management.

[0005] Chinese patent CN101407535B discloses a method for preparing high-purity mogroside V, comprising dissolving it in methanol, filtering it, adding acetone to precipitate the filtrate, subjecting the filtered precipitate to silica gel chromatography, eluting it with a mixed solvent of fatty acid ester and ethanol, and collecting, concentrating, and crystallizing the eluate to obtain mogroside V with a purity of 98% or higher. The process uses organic solvents such as acetone, fatty acid ester, and ethanol, and silica gel chromatography is difficult to reuse due to the small amount of preparation, making large-scale mass production difficult.

[0006] EP2622969A1 discloses a sweetener mixture containing mogroside V with a purity of 60-70%. High-purity mogroside can be obtained by a method of preparing a high-purity mogroside mixture from a low-purity mogroside mixture. Although an innovative multi-column adsorption column system is used in this method, a complex combination of acidic, alkaline, aqueous, and alcoholic solutions is used for impurity elution. The mogroside V content obtained by this method is low, and it fails to provide a method and effect for controlling the content of mogroside II and mogroside III, which are bitter glycosides derived from immature or insufficiently ripe monk fruit raw materials. Furthermore, it fails to provide a method and effect for controlling the protein and sugar content, which affects the application of the product.

[0007] CN104059122A discloses a method for preparing high-purity mogroside V. The core of the method for purifying mogroside V is the innovative use of an unprecedented anion and cation exchange resin with a low particle size of 100-200 mesh (74-149 μm) as a chromatography packing material, and is an atypical and innovative method that uses an aqueous solution with a specific pH gradient to obtain the target product, and does not aim for deionization, but instead uses an ion exchange resin to increase mogroside V. This method does not provide a method or effect for controlling the content of mogroside II and mogroside III, which are bitter glycosides derived from immature or underripe monk fruit raw materials, nor does it provide a method or effect for controlling the protein and sugar content that affects the application of the product.

[0008] To overcome the shortcomings of existing technologies, this application provides a sweet glycoside composition having low protein, low bitterness, low sugar content, and high purity of mogroside V, as well as a method for preparing the same. [Means for solving the problem]

[0009] The specific technical proposal of this application is as follows:

[0010] In one embodiment, the present application is based on the total weight of the sweetening composition, 70%~85% mogroside V, 5% to 12% 11-O-mogroside V, 0-0.5% Siamenoside I, 0-0.05% mogroside II and mogroside III, The present invention provides a sweetening composition containing 0-0.5% protein and 0-0.5% total sugar.

[0011] In another aspect, this application also, Extraction step to obtain an extract from monk fruit; A clarification step is to clarify the extract to obtain a clarified liquid; The ion exchange treatment step involves sequentially treating the clarified liquid with a strongly acidic cation exchange resin column and a weakly basic anion exchange resin column to obtain an ion exchange treatment solution; The ion exchange treatment solution is placed in an adsorption resin column combination and subjected to multi-stage adsorption separation to eluate, the eluate is collected and dried to obtain a sweet composition, and the process is a multi-stage adsorption separation step. The present invention provides a method for preparing a sweetening composition, which includes the following: The ion exchange treatment step must be performed before the multi-stage adsorption separation step.

[0012] Furthermore, in the method for preparing the sweetening composition of this application, the ion exchange treatment solution is sequentially introduced into the 1st, 2nd, ..., nth (n≧2) adsorption resin columns of the adsorption resin column combination to perform multi-stage adsorption separation, and the separated column eluent obtained after passing through each adsorption resin column is either directly introduced into the subsequent adsorption column or The ion exchange treatment solution is sequentially introduced into the 1st, 2nd, ..., nth (n≧2) adsorption resin columns of the adsorption resin column combination to perform multi-stage adsorption separation. After passing through each of the adsorption resin columns from the 1st to the (x-1)th (n≧x≧2) adsorption resin columns, the separated column eluates obtained after passing through each adsorption resin column are collected and combined, then charged into the next adsorption resin column for separation. The column eluates adsorbed and collected in the (x-1)th adsorption resin column are sequentially introduced into the xth, x+1st, x+2nd, ..., nth adsorption resin columns for separation.

[0013] Furthermore, in the method for preparing the sweetening composition of this application, after multi-stage adsorption separation, the sweetening composition of this application is obtained from each of the x~n (2≦x≦n) adsorption resin columns.

[0014] Furthermore, in the method for preparing the sweetening composition of this application, in the multi-stage adsorption separation step, the mass of mogroside V in the ion exchange treatment solution that has passed through the 1st, ..., x-1 (2≦x≦n) adsorption resin columns is 1 to 4 times, preferably 1 to 3 times, and more preferably 1.5 to 3 times, the total adsorbed mass of the 1st to x-1 adsorption resin columns when mogroside V is saturated adsorbed.

[0015] Furthermore, in the method for preparing the sweet composition of the present application, after the clarification step and before the ion exchange treatment step, a step of concentrating mogroside by an adsorption resin column A'' is also included.

[0016] Furthermore, in the method for preparing the sweet composition of the present application, the adsorption resin column A'' is a weakly polar macroporous adsorption resin column. Preferably, the adsorption resin column A'' is a macroporous adsorption resin column whose skeletal material is polyethylene divinylbenzene. Preferably, the particle size of the resin is 250 μm to 1250 μm, preferably 250 μm to 450 μm. Preferably, the resin model of the adsorption resin column A'' is any one selected from LX-15, D101, AB-8, SP700, XD16N, and XD1600N, and preferably SP700.

[0017] Furthermore, in the method for preparing the sweet composition of the present application, the extract is obtained by hot water extraction.

[0018] Furthermore, in the method for preparing the sweet composition of the present application, the clarified liquid is obtained by centrifugation and / or filtration.

[0019] Furthermore, in the method for preparing the sweet composition of the present application, the particle size of the resin of the strongly acidic cation exchange resin column is 300 to 1100 μm, preferably 300 to 600 μm. Preferably, the resin of the strongly acidic cation exchange resin column is one selected from Marathon MSC, AMBERLITE FPC22H, DIAION SK series, and DIAION PK series, and preferably Marathon MSC. Preferably, the particle size of the resin of the weakly basic anion exchange resin column is 350 to 1250 μm, preferably 350 to 600 μm. Preferably, the resin of the weakly basic anion exchange resin column is one selected from Marathon WBA, SCAV2, FPA53 and LX-T5, preferably Marathon WBA or SCAV2.

[0020] Furthermore, in the method for preparing the sweet composition of the present application, each adsorption resin column used in the multi-stage adsorption separation step is a weakly polar macroporous adsorption resin column. Preferably, each of the adsorption resin columns is a macroporous adsorption resin column in which the skeletal material is polyethylene divinylbenzene. Preferably, the resin of each of the adsorption resin columns is one selected from LX-15, D101, AB-8, SP700, XD16N and XD1600N, preferably SP700.

[0021] Furthermore, in the method for preparing the sweet composition of the present application, the multi-stage adsorption separation is a multi-stage adsorption separation using two or more, preferably 2 to 6, adsorption resin columns.

[0022] Furthermore, in the method for preparing the sweet composition of the present application, based on the total weight of the sweet composition, the sweet composition contains 70% to 85% mogroside V, 5% to 12% 11-O-mogroside V, 0 to 0.5% cyanomogroside I, 0 to 0.`05% mogroside II and mogroside III, 0 to 0.5% protein and 0 to 0.5% total sugar.

[0023] <{ Furthermore, in the method for preparing the sweet composition of the present application, in the multi-stage adsorption separation step, after the charging of the ion exchange treatment liquid is completed, the adsorption resin column combination is first eluted with water or organic solvent 1, and then eluted with organic solvent 2. Preferably, the organic solvent 1 is ethanol with a concentration of 10% to 30%. Preferably, the organic solvent 2 is one or two selected from hydrophilic ketones and lower alcohols. Preferably, the organic solvent 2 is one or more selected from methanol, ethanol, propanol, and acetone. Preferably, the organic solvent 2 is ethanol with a concentration of 55% or higher. [Effects of the Invention]

[0024] The method employed in this application first removes ionic substances such as inorganic salts, organic acids, alkaloids, flavones, and proteins from the monk fruit extract using a combination of cation exchange resins and anion exchange resins. Next, it passes through multi-column adsorption resin chromatography, and by controlling the combination and amount of resin in each column, a size exclusion chromatography for adsorbing mogroside components is formed. By utilizing the slight differences in the binding ability of each sweet glycoside component to the resin, the bitter mogroside II and mogroside III first accumulate in the resin of the adsorption resin column, while components with weaker binding ability to the resin, such as 11-O-mogroside V and mogroside V, are pushed into the subsequent series-connected resin columns. Finally, the desired product is obtained by eluting with alcohol of various concentrations. In this application, not only can bitter glycosides be separated and removed, but a more appropriate combination of 11-O-mogroside V and mogroside V can be formed, resulting in a product with a superior taste compared to pure mogroside V. [Brief explanation of the drawing]

[0025] [Figure 1] This is an HPLC chart of various glycoside components in monk fruit according to a specific embodiment of this application. [Figure 2] This is a high-performance liquid chromatogram of the product obtained by eluting the D column with 60% alcohol in Example 5 of this application. [Figure 3] This is a high-performance liquid chromatogram of the product obtained by eluting column E with 60% alcohol in Example 5 of this application. [Modes for carrying out the invention]

[0026] For the sake of ease of understanding, exemplary embodiments of this application are described below, including various details of the embodiments; however, these should be considered merely illustrative. Therefore, those skilled in the art will see that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of this application. Furthermore, for clarity and brevity, well-known descriptions of functions, structures, and reagents are omitted from the following description.

[0027] In one embodiment, the present application provides a sweetening composition, the sweetening composition being based on the total weight of the sweetening composition, 70%~85% mogroside V, 5% to 12% 11-O-mogroside V, 0-0.5% siamenoside I, 0-0.05% mogroside II and mogroside III, It contains 0-0.5% protein and 0-0.5% total sugar.

[0028] The mogroside V of this application is a cucurbitan-type triterpene saponin whose chemical structure is known in the art. In some embodiments, the sweetening composition may contain, based on the total weight of the sweetening composition, for example, 70%, 70.5%, 71%, 71.5%, 72%, 72.5%, 73%, 73.5%, 74%, 74.5%, 75%, 75.5%, 76%, 76.5%, 77%, 77.5%, 78%, 78.5%, 79%, 79.5%, 80%, 80.5%, 81%, 81.5%, 82%, 82.5%, 83%, 83.5%, 84%, 84.5%, 85%, or any range in between.

[0029] The 11-O-mogroside V of this application is a cucurbitan-type triterpene saponin, a sweetener whose chemical structure is known in the art, possesses potent antioxidant activity, and exhibits a significant inhibitory effect on reactive oxygen species. In some embodiments, the sweetening composition may contain 11-O-mogroside V in amounts of, for example, 5%, 5.2%, 5.5%, 5.8%, 6%, 6.2%, 6.5%, 6.7%, 7%, 7.3%, 7.5%, 7.7%, 8%, 8.2%, 8.5%, 8.8%, 9%, 9.2%, 9.5%, 9.8%, 10%, 10.3%, 10.5%, 10.8%, 11%, 11.2%, 11.5%, 11.7%, 12%, or any range between these amounts, based on the total weight of the sweetening composition.

[0030] Siamenoside I is a cucurbitan-type triterpene saponin and the sweetest component among mogrosides, with a sweetness approximately 460 to 563 times that of sucrose, although it is naturally present in very small amounts in monk fruit. In some embodiments, the sweetening composition is expressed in amounts based on the total weight of the sweetening composition, for example, 0%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, It may contain siamenoside I in any range between 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, or any range between them.

[0031] Mogrosides typically have a varying number (2-6) of glucose units linked to carbon 3 and carbon 24 of the triterpene backbone. Mogroside II is the simplest form of mogroside, with one glucose residue linked to both carbon 3 and carbon 24. Mogroside III differs in that an additional glucose residue is linked to carbon 24. In some embodiments, the total weight of mogroside II and mogroside III in the sweetening composition may be, for example, 0%, 0.001%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, or any range in between, of the total weight of the sweetening composition.

[0032] In some embodiments, the sweetening composition may contain protein in any range between 0%, 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or any other range based on the total weight of the sweetening composition. The protein content in this application can be measured by any suitable method or technique known in the art, for example, by the first method of GB5009.5-2016.

[0033] In some embodiments, the sweetening composition may contain total sugars in any range between 0%, 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or any other range based on the total weight of the sweetening composition. In this application, total sugar content refers to the sum of monosaccharide and disaccharide content based on dry weight. In this application, total sugar content can be measured by any suitable method or technique known in the art, for example, by the first method of GB5009.8-2016, which can be detected using HPLC-RID.

[0034] In another aspect, this application also, Extraction step to obtain an extract from monk fruit; A clarification step is to clarify the extract to obtain a clarified liquid; The ion exchange treatment step involves sequentially treating the clarified liquid with a strongly acidic cation exchange resin column and a weakly basic anion exchange resin column to obtain an ion exchange treatment solution; The ion exchange treatment solution is placed in an adsorption resin column combination and subjected to multi-stage adsorption separation to eluate, the eluate is collected and dried to obtain a sweet composition, and the process is a multi-stage adsorption separation step. The present invention provides a method for preparing a sweetening composition, which includes the following: The ion exchange treatment step must be performed before the multi-stage adsorption separation step.

[0035] This application investigates the differences in the selectivity of adsorption of mogroside V, other mogroside components, and other components in macroporous adsorption resins for monk fruit, based on a conventional mogroside manufacturing process, namely a process combining mogroside-grinding-water extraction-adsorption on macroporous resins and decolorization of the resin. As a result, it was found that when ion-exchange treated monk fruit extract is passed through a column composed of multiple macroporous resin columns, the distribution of mogroside V, other mogroside components, and other components in each column can be adjusted by controlling the amount of charge. By utilizing the elution and drying of each column, sweet compositions containing different combinations of mogrosides can be obtained, and the mogroside V content in the sweet compositions can reach 70% to 85%.

[0036] In some embodiments, the method for preparing the sweetening composition of this application includes contacting monk fruit with water to prepare an extract containing mogroside V, other terpene glycosides and sugars; clarifying the extract to remove pectin and proteins to facilitate subsequent processing; sequentially contacting the clarified liquid with a strongly acidic cation exchange resin column and a weakly basic anion exchange resin column to decolorize, deacidify, and further deproteinize; contacting the ion-exchanged liquid obtained after ion exchange treatment with a combination of adsorption resin columns to elute, maximizing the content of mogroside V, 11-O-mogroside V, and siamenoside I in the eluate while minimizing the content of mogroside II, mogroside III, and proteins; and finally drying to obtain the sweetening composition of this application.

[0037] In some embodiments, the extract is obtained by extraction with hot water. The temperature of the hot water may be, for example, 70°C, 72°C, 74°C, 76°C, 78°C, 80°C, 82°C, 84°C, 86°C, 88°C, 90°C, 92°C, 94°C, 96°C, 98°C, 100°C, etc.

[0038] In some embodiments, the extraction step involves crushing fresh or dried monk fruit and extracting it with hot water to obtain an extract.

[0039] In this application, clarification can be carried out using any suitable method known in the art, such as centrifugation and / or filtration, preferably by ultrafiltration, for example, using an ultrafiltration membrane. Alternatively, clarification can be carried out by treatment with phosphoric acid or pectinase. Pectinase in the form of a commercially available enzyme mixture containing pectinase can be used to decompose pectin from the extract and precipitate pectin-stabilized peptides and proteins.

[0040] In some embodiments, the extract is initially clarified by centrifugation in a centrifuge, and the supernatant is further clarified by ultrafiltration through a ceramic membrane to obtain a clarified liquid.

[0041] The method of this application uses an ion exchange resin to remove inorganic salts, proteins, amino acids, and charged components from a clarified liquid obtained from monk fruit, while simultaneously retaining the maximum amount of specific terpene glycosides, particularly mogroside V, which helps to obtain a sweet composition with a pure flavor. An ion exchange resin is a polymer that allows specific ions within the polymer to be exchanged with ions in the solution passing through the resin. Since ion exchange resins can be insoluble acids or bases, and their salts are also insoluble, the resin can exchange positively charged ions (cation exchange resins) or negatively charged ions (anion exchange resins). In this application, the strong sweetness of the sweet composition can be retained to the maximum extent by performing the ion exchange treatment using a strongly acidic cation exchange resin column and a weakly basic anion exchange resin column in sequence. It should be understood that the ion exchange resins used in the method described herein can be obtained from any source, including any commercially available source.

[0042] In some embodiments, the compound responsible for the grassy or earthy taste or smell, and bitterness, is a bitter melanoidin. In some embodiments, the compound is a bitter peptide. In some embodiments, the compound is a bitter terpenoid. In some embodiments, the compound is picrol. In some embodiments, the compound is a bitter polyphenol. In some embodiments, the compound is a bitter phenolic oligomer. In some embodiments, the compound is a bitter condensed polyphenol. In some embodiments, the compound is a bitter terpene glycoside.

[0043] In some embodiments, the resin particle size of the strongly acidic cation exchange resin column is 300 to 1100 μm, and the resin particle size of the weakly basic anion exchange resin column is 350 to 1250 μm.

[0044] In some embodiments, the resin particle size of the strongly acidic cation exchange resin column is 300 to 600 μm, and the resin particle size of the weakly basic anion exchange resin column is 350 to 600 μm.

[0045] In some embodiments, the resin for the strongly acidic cation exchange resin column is selected from Marathon MSC, AMBERLITE FPC22H, DIAION SK series, and DIAION PK series, and the resin for the weakly basic anion exchange resin column is selected from Marathon WBA, SCAV2, FPA53, and LX-T5.

[0046] In some embodiments, the resin of the strongly acidic cation exchange resin column is Marathon MSC, and the resin of the weakly basic anion exchange resin column is Marathon WBA or SCAV2.

[0047] Typically, based on the selectivity of macroporous adsorption resins, in common water and alcohol solvent systems, mogroside IV, siamenoside I, mogroside II, mogroside III, mogrol, and other non-sweet glycoside components, which are less polar than mogroside V, are more strongly retained on the resin than mogroside V. Conversely, components that are more polar than mogroside V, such as 11-O-mogroside V and other non-sweet glycoside components, exhibit weaker retention than mogroside V. These differences in resin selectivity are not evident in typical single-column resin adsorption and are difficult to fully utilize. However, when using a multi-column combination, components in the material accumulate and are amplified based on differences in polarity or adsorption selectivity, forming an effect similar to exclusion chromatography. When charged into a resin column, less polar components and components with relatively strong adsorption capacity gradually accumulate in the earlier columns. If the amount of charged material is sufficient, these components occupy the adsorption capacity of the earlier resin columns, while mogroside V and other more polar components, having weaker competitive adsorption capacity, are eluted or pushed into subsequent chromatography columns. Components entering the later columns also differ in polarity and resin adsorption capacity. Components more polar than mogroside V are competitively adsorbed by mogroside V and pushed further into the later columns or the eluent. As a result, there are significant differences in the composition of components and the amount of mogroside V in adsorption columns arranged in different configurations.

[0048] In some embodiments, the ion exchange treatment solution is sequentially introduced into the 1st, 2nd, ..., nth (n≧2) adsorption resin columns of the adsorption resin column combination to perform multi-stage adsorption separation. Here, after passing through each of the adsorption resin columns from the 1st to the (x-1)th (n≧x≧2) adsorption resin columns, the separated column phosphate obtained after passing through each adsorption resin column is collected and combined, then charged into the next adsorption resin column for separation. The column phosphate adsorbed and collected in the (x-1)th adsorption resin column is sequentially introduced into the xth, x+1th, x+2nd, ..., nth adsorption resin columns for adsorption separation. In some embodiments, the ion exchange treatment solution is sequentially introduced into the 1st, 2nd, ..., nth (n≧2) adsorption resin columns of the adsorption resin column combination to perform multi-stage adsorption separation. Here, the separated column phosphate obtained after passing through each adsorption resin column is not collected but directly entered into the next adsorption column. In series separation, the separated components are distributed sequentially within the resin layer according to the difference in the bonding strength between the molecules and the resin (strongly adsorbed substances flow out later, and weakly adsorbed substances flow out earlier). When the fractions are collected between columns and combined, the order of the components separated in the previous column is disrupted again, making separation in subsequent columns even more difficult. Therefore, in the two embodiments described above, the latter has a superior adsorption separation effect than the former. In the method of this application, after multi-stage adsorption separation, the sweetening composition of this application is obtained from each of the x~n (2≦x≦n) adsorption resin columns.

[0049] In some embodiments, the multistage adsorption separation in this application is a multistage adsorption separation using two or more, preferably two to six, adsorption resin columns, for example, two, three, four, five, or six adsorption resin columns, and more preferably two, three, or four adsorption resin columns. In this application, the resin models of each adsorption resin column are the same or different, and the resin amounts of each adsorption resin column are the same or different.

[0050] In some embodiments, in the multi-stage adsorption separation step, the mass of mogroside V in the ion exchange solution that has passed through the 1st, ..., x-1 (2≦x≦n) adsorption resin columns is 1 to 4 times the total adsorbed mass of the 1st to x-1 adsorption resin columns when mogroside V is saturated adsorbed, for example, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, and so on, preferably 1 to 3 times, more preferably 1.5 to 3 times. The multiplier is relative to the volume of resin in the column that cannot achieve high purity. If the multiplier is too high, there will be a large amount of strongly adsorbed impurities (weakly polar substances, mogroside II, mogroside III, etc.), which will be pushed out into column x beyond the adsorption and retention capacity of column x, increasing the amount of impurities in column x and affecting the purity and taste of the purified sweet glycosides. If the amount of material used is too small, the resin utilization rate of column x-1 will be too low, resulting in too few sweet glycosides entering columns x-n. While this may yield a higher purity product, the yield will be too low, resulting in very low economic benefits.

[0051] In some embodiments, a mogroside concentration step using an adsorption resin column A'' is also included after the clarification step and before the ion exchange treatment step. In some embodiments, the mogroside concentration step includes concentrating the mogroside by adsorbing the clarified liquid with a macroporous adsorption resin, eluting with alcohol to obtain a mogroside eluate, concentrating and separating the alcohol to obtain a mogroside concentrate, diluting and filtering to obtain a filtrate for use in the subsequent ion exchange treatment step.

[0052] In some embodiments, during the concentration step, the adsorption resin column A'' is a macroporous adsorption resin column whose backbone material is polyethylenedivinylbenzene, and the particle size of the resin is 250 μm to 1250 μm.

[0053] In some embodiments, the resin model of the adsorption resin column A'' in the concentration step includes, but is not limited to, commonly used macroporous adsorption resins such as LX-15, D101, AB-8, SP700, XD16N, and XD1600N, and is preferably SP700.

[0054] In some embodiments, each adsorption resin column used in the multi-stage adsorption separation step is a weakly polar macroporous adsorption resin column, preferably each adsorption resin column A is a macroporous adsorption resin column whose backbone material is polyethylenedivinylbenzene, and the resin of each adsorption resin column includes, but is not limited to, commonly used macroporous adsorption resins such as LX-15, D101, AB-8, SP700, XD16N, and XD1600N, but is preferably SP700.

[0055] In some embodiments, the sweetening composition may contain mogroside V in any range between these amounts, based on the total weight of the sweetening composition, for example, 70%, 70.5%, 71%, 71.5%, 72%, 72.5%, 73%, 73.5%, 74%, 74.5%, 75%, 75.5%, 76%, 76.5%, 77%, 77.5%, 78%, 78.5%, 79%, 79.5%, 80%, 80.5%, 81%, 81.5%, 82%, 82.5%, 83%, 83.5%, 84%, 84.5%, 85%, or any range between these amounts.

[0056] In some embodiments, the sweetening composition may contain 11-O-mogroside V in amounts based on the total weight of the sweetening composition, for example, 5%, 5.2%, 5.5%, 5.8%, 6%, 6.2%, 6.5%, 6.7%, 7%, 7.3%, 7.5%, 7.7%, 8%, 8.2%, 8.5%, 8.8%, 9%, 9.2%, 9.5%, 9.8%, 10%, 10.3%, 10.5%, 10.8%, 11%, 11.2%, 11.5%, 11.7%, 12%, or any range between these amounts.

[0057] In some embodiments, the sweetening composition is expressed in amounts based on the total weight of the sweetening composition, for example, 0%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, It may contain siamenoside I in any range between 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, or any range between them.

[0058] In some embodiments, the total weight of mogroside II and mogroside III contained in the sweetening composition may be, for example, 0%, 0.001%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, or any range in between, of the total weight of the sweetening composition.

[0059] In some embodiments, the sweetening composition may contain protein in any range between 0%, 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or any range between these, based on the total weight of the sweetening composition.

[0060] In some embodiments, the sweetening composition may contain, based on the total weight of the sweetening composition, total sugars of, for example, 0%, 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or any range in between.

[0061] In some embodiments, in the multi-stage adsorption separation step, after the preparation of the ion exchange treatment solution is complete, the adsorption resin column combination is first eluted with water or 10% to 30% ethanol, and then eluted with one or more solvents selected from methanol, ethanol, propanol, and acetone.

[0062] In some embodiments, in the multi-stage adsorption separation step, after the preparation of the ion exchange treatment solution is complete, the adsorption resin column combination is first eluted with 30% ethanol, and then eluted with 55% or more ethanol.

[0063] The sweetening compositions described herein can be used in food, beverage, pharmaceutical, or dietary supplement products. [Examples]

[0064] This application provides a general and / or specific description of the materials and test methods used in the tests. In the following examples, the materials or equipment used are conventional materials or equipment that are commercially available unless the manufacturer is indicated.

[0065] Example 1 300 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0066] The ultrafiltrate is processed using a macroporous resin (model SP700, particle size 250 μm, specific surface area 1200 m²). 2The sample was placed in a chromatography column (30 cm in diameter x 90 cm in height) equipped with 50 L of ( / g) of mogrosides, adsorbed, and the column was washed with 200 L of purified water. Next, it was eluted with 150 L of 60% alcohol, the alcohol eluate was collected and concentrated under reduced pressure to obtain alcohol-free sweet glycoside-containing concentrate b. The net weight was 13.8 kg, the solid content of the concentrate was 30%, and the mogroside V content in the concentrate, converted to dry matter, was 26.5%. The concentrated solution is diluted with purified water to a solid content of 5% and subjected to ion exchange treatment. The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, with a particle size of 550-900 μm. The weakly basic anion exchange resin model is DOWEX Marathon WBA, with a particle size of 475-575 μm. The resin capacity of each column is 15 L. Diluted concentrated solutions are sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are washed with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 2.8%, the volume is 65.9 L, and the mogroside V content in the solution, converted to dry matter, is 59.6%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of three resin columns, each packed with 6L of macroporous adsorption resin (SP700), and is numbered A, B, and C in order. All columns are regenerated, washed with water, and prepared for use.

[0067] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, and C at a flow rate of 3 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange treatment solution after passing through column A was 2.6 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B and C were eluted with 12 L of 25% alcohol each. Then, each column was eluted with 60% ethanol, and the eluates were collected separately, concentrated, and dried to detect the content of mogroside V and other glycoside components. The results are shown in Table 1.

[0068] [Table 1]

[0069] Example 2 356 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor and extracted with 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain 1200 L of ultrafiltrate a. This was then subjected to ion exchange treatment. The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 120 L for each column. Diluted concentrated solutions are sequentially supplied to the cation exchange resin column and the anion exchange resin column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, with a particle size of 550-900 μm. The weakly basic anion exchange resin model is DOWEX Marathon WBA, with a particle size of 475-575 μm. After the setup is complete, the columns are eluted with purified water, and the fluid flowed through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 1.92%, the volume is 1300 L, and the mogroside V content, converted to dry matter in the solution, is 5.4%. This solution is then used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each packed with 5L of macroporous adsorption resin. The resin model is SP700, with a particle size of 250μm and a specific surface area of ​​1200m². 2 The quantity is per g, and the columns are numbered A, B, C, and D in order. All are regenerated, washed with water, and prepared for use.

[0070] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange treatment solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was washed with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were each eluted with 12 L of 25% alcohol. Then, each column was eluted with 10 L of 60% ethanol, and the eluates were collected separately, concentrated, and dried. The content of mogroside V and other glycoside components was detected. The results are shown in Table 2.

[0071] [Table 2]

[0072] Example 3 370 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a. This was then subjected to ion exchange treatment. The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 130 L for each column. The strongly acidic cation exchange resin model is AMBERLITE FPC3500, with a particle size of 450-650 μm. The weakly basic anion exchange resin model is SCAV2, with a particle size of 600-800 μm. Ultrafiltrate a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are washed with purified water, and the fluid flowed through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 1.7%, the volume is 1200 L, and the mogroside V content, converted to dry matter in the solution, is 5.5%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of three resin columns, each packed with 8L of macroporous adsorption resin. The resin model is XAD16N, with a particle size of 560-710 μm and a specific surface area of ​​800 m². 2 The result was / g, and preliminary tests showed that the glycoside adsorption capacity of saturated XAD16N unit resin was 6.3g / 100ml.

[0073] The resin columns are numbered A, B, and C in order, and all are regenerated, washed with water, and prepared for use.

[0074] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, and C at a flow rate of 2.4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange treatment solution after passing through column A was three times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 2.5 BV purified water, then eluted with 16 L of 25% alcohol, and finally eluted with 16 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycoside components was detected. The results are shown in Table 3.

[0075] [Table 3]

[0076] Example 4 264 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 900 L of 85°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a. The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 92 L for each column. The strongly acidic cation exchange resin model is AMBERLITE FPC3500, with a particle size of 450-650 μm. The weakly basic anion exchange resin model is SCAV2, with a particle size of 600-800 μm. Diluted concentrated solutions are sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowed through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 1.8%, the volume is 12.0 L, and the mogroside V content, converted to dry matter in the solution, is 5.5%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of two resin columns (model XAD1600N) packed with 6L and 8L of macroporous adsorption resin, respectively. The particle size is 350-450 μm, and the specific surface area is 800 m². 2 They are labeled A and B in order, and both are recycled, washed with water, and prepared for use.

[0077] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A and B at a flow rate of 1.8 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange treatment solution after passing through column A was 2.7 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 3 BV of purified water, then with 2 BV of 25% alcohol, and finally with 2 BV of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 4.

[0078] [Table 4]

[0079] Example 5 300 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0080] The ultrafiltrate is processed using a macroporous resin (Model LX-15, particle size 315-1250 μm, specific surface area 750 m²). 2 The sample was placed in a chromatography column (30 cm in diameter x 90 cm in height) equipped with 45 L of ( / g) of mogrosides, adsorbed, and the column was eluted with 200 L of purified water. Next, it was eluted with 150 L of 60% alcohol, and the alcohol eluate was collected and concentrated under reduced pressure to obtain alcohol-free sweet glycoside-containing concentrate b. The net weight was 13.6 kg, the solid content of the concentrate was 35%, and the mogroside V content in the concentrate, converted to dry matter, was 25.8%. The concentrated solution is diluted with purified water to a solid content of 5% and subjected to ion exchange treatment. The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, with a particle size of 550-900 μm. The weakly basic anion exchange resin model is SCAV2, with a particle size of 600-800 μm. The resin capacity of each column is 5.5 L. Diluted concentrated solutions are sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowed through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 3.0%, the volume is 63.5 L, and the mogroside V content, converted to dry matter in the solution, is 58.0%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of five adsorption resin columns numbered A, B, C, D, and E in order. Column A is filled with 5.5 L of Model SP700 resin, Column B with 4.5 L of Model XAD1600N resin, and Columns C, D, and E with 2.25 L of XAD1600N resin. All columns are regenerated with a 5% sodium hydroxide solution and washed with water before use.

[0081] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, D, and E at a flow rate of 1.2 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was 1.7 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted and purified with 2 BV of purified water. Next, columns A and B were directly eluted with 60% ethanol, and columns C, D, and E were eluted with 5 L of 25% alcohol each, followed by 5 L of 60% ethanol each. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, protein content, and total sugar content was detected. The results are shown in Table 5.

[0082] [Table 5]

[0083] Example 6 330 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0084] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 115 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, with a particle size of 450-650 μm. The weakly basic anion exchange resin model is SCAV2, with a particle size of 600-800 μm. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the preparation is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.1%, the volume is 1212 L, and the mogroside V content in the solution, converted to dry matter, is 5.5%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0085] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were eluted with 25% alcohol of 2 BV each, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content was detected. The results are shown in Table 6.

[0086] [Table 6]

[0087] Example 7 356 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0088] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 125 L for each column. The strongly acidic cation exchange resin model is AMBERLITE FPC22H, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid obtained from flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 2.3%, the volume is 1210 L, and the mogroside V content in the solution, converted to dry matter, is 5.4%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0089] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were eluted with 25% alcohol of 2 BV each, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content was detected. The results are shown in Table 7.

[0090] [Table 7]

[0091] Example 8 360 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0092] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 126 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is LX-T5. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.1%, the volume is 1370 L, and the mogroside V content in the solution, converted to dry matter, is 5.3%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0093] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were each eluted with 25% alcohol of 2 BV, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content was detected. The results are shown in Table 8.

[0094] [Table 8]

[0095] Example 9 361 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0096] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 130 L for each column. The strongly acidic cation exchange resin model is AMBERLITE FPC22H, and the weakly basic anion exchange resin model is FPA53. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.2%, the volume is 1238 L, and the mogroside V content in the solution, converted to dry matter, is 5.6%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0097] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were eluted with 25% alcohol of 2 BV each, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content was detected. The results are shown in Table 9.

[0098] [Table 9]

[0099] Example 10 366 kg of fresh monk fruit were finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0100] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 130 L for each column. The strongly acidic cation exchange resin model is FPC88 UPS Na, and the weakly basic anion exchange resin model is FPA66 UPS. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 2.3%, the volume is 1247 L, and the mogroside V content in the solution, converted to dry matter, is 5.4%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0101] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were eluted with 25% alcohol of 2 BV each, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content was detected. The results are shown in Table 10.

[0102] [Table 10]

[0103] Example 11 110 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 300 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0104] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 39 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 2.3%, the volume is 320 L, and the mogroside V content in the solution, converted to dry matter, is 5.7%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of two resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A and B, and all are regenerated, washed with water, and prepared for use.

[0105] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A and B at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange solution after passing through column A was 1.2 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. The two columns were eluted with 3 BV of purified water. Next, column A was directly eluted with 60% ethanol, and the other column was eluted with 2 BV of 25% alcohol, followed by 2 BV of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 11.

[0106] [Table 11]

[0107] Example 12 135 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 300 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0108] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 47 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.5%, the volume is 396 L, and the mogroside V content in the solution, converted to dry matter, is 5.3%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of two resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A and B, and all are regenerated, washed with water, and prepared for use.

[0109] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A and B at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange solution after passing through column A was 1.5 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. The two columns were eluted with 3 BV of purified water. Next, column A was directly eluted with 60% ethanol, and the other column was eluted with 2 BV of 25% alcohol, followed by 2 BV of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 12.

[0110] [Table 12]

[0111] Example 13 200 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 600 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0112] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 70 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.5%, the volume is 625 L, and the mogroside V content in the solution, converted to dry matter, is 5.3%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of three resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, and C in order, and all are regenerated, washed with water, and prepared for use.

[0113] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, and C at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange treatment solution after passing through column A was 2.2 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 3 BV of purified water. Next, column A was directly eluted with 60% ethanol, and the other columns were eluted with 2 BV of 25% alcohol, followed by 2 BV of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 13.

[0114] [Table 13]

[0115] Example 14 270 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 600 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0116] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 95 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.4%, the volume is 754 L, and the mogroside V content in the solution, converted to dry matter, is 5.8%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of three resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, and C in order, and all are regenerated, washed with water, and prepared for use.

[0117] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, and C at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchange treatment solution after passing through column A was three times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 3 BV of purified water. Next, column A was directly eluted with 60% ethanol, and the other columns were eluted with 2 BV of 25% alcohol, followed by 2 BV of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 14.

[0118] [Table 14]

[0119] Example 15 565 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 900 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0120] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 200 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.35%, the volume is 1820 L, and the mogroside V content in the solution, converted to dry matter, is 5.73%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0121] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 5 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was 3.5 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 3 BV of purified water. Next, column A was directly eluted with 60% ethanol, and the other columns were eluted with 2 BV of 25% alcohol, followed by 2 BV of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 15.

[0122] [Table 15]

[0123] Example 16 282 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0124] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 100 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.1%, the volume is 918 L, and the mogroside V content in the solution, converted to dry matter, is 5.5%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model AB-8). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0125] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were eluted with 25% alcohol of 2BV, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 16.

[0126] [Table 16]

[0127] Example 17 263 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0128] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 92 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are washed with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content of this solution is 2.1%, the volume is 866 L, and the mogroside V content in the solution, converted to dry matter, is 5.5%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (Model D101). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0129] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12.5 L of purified water. Next, column A was directly eluted with 60% ethanol, and columns B, C, and D were eluted with 25% alcohol of 2BV, followed by elution with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 17.

[0130] [Table 17]

[0131] Example 18 360 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 900 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0132] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 126 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.4%, the volume is 1005 L, and the mogroside V content in the solution, converted to dry matter, is 5.8%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of five resin columns, each filled with 4L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, D, and E in order, and all are regenerated, washed with water, and prepared for use.

[0133] The above deionized solution c was sequentially passed through macroporous adsorption resin columns A, B, C, D, and E at a flow rate of 4 L / h. After the flow was complete, the total amount of mogroside V in the ion-exchanged solution after passing through columns A and B was 2.5 times the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12 L of purified water. Next, column A was directly eluted with 10 L of 60% ethanol. Columns B, C, D, and E were first eluted with 2BV of 25% alcohol, and then eluted with 10 L of 60% ethanol. The eluates were collected separately, concentrated, and dried, and the content of mogroside V and other glycosides, protein content, and total sugar content was detected. The results are shown in Table 18.

[0134] [Table 18]

[0135] Example 19 360 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 900 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0136] The ion exchange resin column consists of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 126 L for each column. The strongly acidic cation exchange resin model is DOWEX Marathon MSC, and the weakly basic anion exchange resin model is DOWEX Marathon WBA. Solution a is sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup is complete, the columns are eluted with purified water, and the fluid flowing through the ion exchange resin-treated columns is combined with the eluted aqueous solution to obtain a mixed deionized solution c. The solid content is 2.4%, the volume is 1005 L, and the mogroside V content in the solution, converted to dry matter, is 5.8%. This solution is used for multi-stage adsorption separation and purification. The multi-stage adsorption separation column group consists of four resin columns, each filled with 5L of macroporous adsorption resin (model SP700). The columns are numbered A, B, C, and D in order, and all are regenerated, washed with water, and prepared for use.

[0137] The above deionizing solution c was passed through column A at a flow rate of 5 L / h, the column solution was collected, and then passed through column B at a flow rate of 5 L / h, and the column solution was collected. Next, the column solutions were sequentially passed through macroporous adsorption resin columns C and D. After the passage was complete, the total amount of mogroside V in the ion exchange treatment solution after passing through columns A and B was twice the saturation adsorption amount of mogroside V based on the corresponding amount of resin. Each column was eluted with 12 L of purified water. Next, column A was directly eluted with 10 L of 60% ethanol, and columns B, C, and D were eluted with 2 BV of 25% alcohol, and then eluted with 10 L of 60% ethanol. The eluates were collected separately, concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 19.

[0138] [Table 19]

[0139] Comparative Example 1 300 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor, and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0140] The ultrafiltrate is processed using a macroporous resin (model SP700, particle size 250 μm, specific surface area 1200 m²). 2 The sample was placed in a chromatography column (30 cm in diameter x 90 cm in height) equipped with 50 L of ( / g) of mogrosides, adsorbed, and the column was eluted with 200 L of purified water. Next, it was eluted with 150 L of 60% alcohol, and the alcohol eluate was collected and concentrated under reduced pressure to obtain alcohol-free sweet glycoside-containing concentrate b. The net weight was 13.8 kg, the solid content of the concentrate was 30%, and the mogroside V content in the concentrate, converted to dry matter, was 26.5%. The concentrated solution is diluted with purified water to a solid content of 5% and subjected to a multi-stage adsorption separation process. The multi-stage adsorption separation column group consists of three resin columns, each packed with 6 L of macroporous adsorption resin. The resin model is SP700, and the columns are numbered A, B, and C in order. All are regenerated and washed with water to prepare for use. The ultrafiltrate a described above is passed sequentially through macroporous adsorption resin columns A, B, and C at a flow rate of 3000 ml / h. After the flow is complete, each column is eluted with 12 L of purified water. Next, column A is directly washed with 60% ethanol, and columns B and C are eluted with 12 L of 25% alcohol each. Then, each column is eluted with 60% ethanol, and the eluates are collected separately and concentrated to obtain 60% alcohol eluates for columns A, B, and C. Each concentrate is diluted with purified water to a Brix concentration of approximately 5.0%, and each is subjected to ion exchange treatment.

[0141] The ion exchange resin column consisted of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 1.8 L for each column. Diluted concentrated solutions were sequentially supplied to the cation exchange resin column and the anion exchange resin column. The strongly acidic cation exchange resin model was DOWEX Marathon MSC, and the weakly basic anion exchange resin model was DOWEX Marathon WBA. After preparation, the columns were eluted with purified water, and the fluid flowed through the ion exchange resin-treated columns was combined with the eluted aqueous solution to obtain a mixed deionized solution. The deionized solutions from each column were concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 20.

[0142] [Table 20]

[0143] Comparative Example 2 300 kg of fresh monk fruit were finely crushed in a pulverizer, then placed in a countercurrent extractor and extracted with 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain 1200 L of ultrafiltrate a. This was then subjected to a multi-stage adsorption separation process. The multi-stage adsorption separation column group consists of four resin columns, each packed with 5L of macroporous adsorption resin. The resin model is SP700, and the columns are numbered A, B, C, and D in order. All are regenerated and washed with water to prepare for use. The ultrafiltrate a described above is passed sequentially through macroporous adsorption resin columns A, B, C, and D at a flow rate of 3900 ml / h. After the flow is complete, each column is eluted with 10L of purified water. Next, column A is directly washed with 60% ethanol, and columns B, C, and D are eluted with 10L of 25% alcohol each. Then, each column is eluted with 60% ethanol, and the eluates are collected separately and concentrated to obtain 60% alcohol eluates for columns A, B, C, and D. Each concentrate is diluted with purified water to a Brix concentration of approximately 5.0%, and each is subjected to ion exchange treatment.

[0144] The ion exchange resin column consisted of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 1.5 L for each column. Diluted concentrated solutions were sequentially supplied to the cation exchange resin column and the anion exchange resin column. The strongly acidic cation exchange resin model was DOWEX Marathon MSC, and the weakly basic anion exchange resin model was DOWEX Marathon WBA. After preparation, the columns were eluted with purified water, and the fluid flowed through the ion exchange resin-treated columns was combined with the eluted aqueous solution to obtain a mixed deionized solution. The deionized solutions from each column were concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 21.

[0145] [Table 21]

[0146] Comparative Example 3 280 kg of fresh monk fruit was finely crushed in a pulverizer, then placed in a countercurrent extractor and extracted using 1000 L of 90°C hot water. The extract was processed in a press, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a. This was then subjected to a multi-stage adsorption separation process.

[0147] The multi-stage adsorption separation column group consists of three resin columns, each packed with 8L of macroporous adsorption resin. The resin model is XAD16N, and the columns are numbered A, B, and C in order. All are regenerated and washed with water to prepare for use. The ultrafiltrate a described above is passed sequentially through macroporous adsorption resin columns A, B, and C at a flow rate of 3200 ml / h. After the flow is complete, each column is eluted with 16L of purified water. Next, column A is directly washed with 60% ethanol, and columns B and C are eluted with 16L of 25% alcohol each. Then, each column is eluted with 60% ethanol, and the eluates are collected separately and concentrated to obtain 60% alcohol eluates for columns A, B, C, and D. Each concentrate is diluted with purified water to a Brix concentration of approximately 5.0%, and each is subjected to ion exchange treatment.

[0148] The ion exchange resin column consisted of one strongly acidic cation exchange resin column and one weakly basic anion exchange resin column connected in series, with a resin capacity of 1.8 L for each column. The strongly acidic cation exchange resin model was AMBERLITE FPC3500, and the weakly basic anion exchange resin model was SCAV2. Ultrafiltrate a was sequentially supplied to the cation exchange resin column and the anion exchange resin column. After the setup was complete, the columns were eluted with purified water, and the fluid flowed through the ion exchange resin-treated columns was combined with the eluted aqueous solution to obtain a mixed deionized solution. The deionized solutions from each column were concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were detected. The results are shown in Table 22.

[0149] [Table 22]

[0150] The experimental parameters, conditions, and effect data for each example and comparative example are shown in Table 23 below.

[0151] [Table 23A]

[0152] [Table 23B]

[0153] [Table 23C]

[0154] Comparing the above examples with comparative examples, it can be seen that when the monk fruit extract was subjected to multi-stage adsorption separation directly without ion exchange treatment, a sweet glycoside composition with more than 70% mogroside V could not be obtained. This indicates that in materials that have not undergone ion exchange treatment, the components are too complex, affecting the separation efficiency of the adsorption resin, and that an ion exchange treatment step is necessary before adsorption separation.

[0155] In this application, the extraction effect of mogroside V is comprehensively evaluated from two perspectives: the purity of the obtained product and the proportion of high-purity products. The higher the purity of the obtained product and the greater the proportion of high-purity products, the greater the effect. Comparing the separation and purification effects of each example and comparative example, when the content of mogroside V separated in the last resin column is similar, the greater the total net weight percentage of mogroside V obtained from each resin column where the separated mogroside V content is 70% or more, the greater the separation and purification effect. When the total net weight percentage of mogroside V obtained from each resin column where the separated mogroside V content is 70% or more is similar, the greater the content of mogroside V separated in the last resin column, the greater the separation and purification effect.

[0156] Experimental example 1. Method for detecting each component of monk fruit products 1.1 Method for detecting the content of mogroside V Mogroside V was detected according to the method corresponding to national standard GB 1886.77-2016. 1.2 Method for detecting the content of other glycoside components in monk fruit Mogroside components were detected by high-performance liquid chromatography using an external standard method. The chromatography conditions and sample handling procedure are as follows. 1.2.1 Chromatography Conditions 1.2.1.1 Chromatography column: ZORBAX SB-C18, 4.6 × 250 mm, 5 μm. 1.2.1.2 The mobile phase and its corresponding gradient are shown in Table 24.

[0157] [Table 24]

[0158] 1.2.1.3 Detector: Diode array UV detector 1.2.1.4 Detection wavelength: 203nm 1.2.1.5 Column temperature: 30℃ 1.2.1.6 Preparation volume: 20 μL 1.2.2 Sample Operation Method 1.2.2.1 Siamenoside I, mogroside IIe, mogroside III, mogroside IVe, 11-O-mogroside V, and mogroside V control substances were accurately weighed. Siamenoside I, mogroside IIe, mogroside III, and mogroside IVe were dissolved in purified water to prepare four standard solutions at concentrations of 0.015 mg / ml, 0.03 mg / ml, 0.045 mg / ml, and 0.075 mg / ml. Standard solutions with concentrations of 11-O-mogroside V of 0.03 mg / ml, 0.06 mg / ml, 0.09 mg / ml, and 0.15 mg / ml, respectively, and standard solutions with concentrations of mogroside V of 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, and 0.5 mg / ml were prepared. Figure 1 shows the HPLC spectra of various glycoside components in monk fruit, obtained by detecting the prepared samples according to the above chromatography conditions. Peak area data was collected to create external calibration curves for each control substance. 1.2.2.2 An appropriate amount of sample was taken, accurately weighed, dissolved in ultrapure water, and made up to prepare a sample solution of approximately 3 mg / ml. Liquid chromatography detection was performed according to the above chromatographic conditions, a chromatogram was collected, and the components in the sample were identified using the retention time corresponding to each standard. Quantitative analysis was performed according to the corresponding calibration curve to calculate the content of each component. 1.3 Method for detecting the protein content of monk fruit products The method for detecting the protein content of monk fruit products in this application employs the first method described in GB 5009.5-2016. 1.4 Method for detecting the total sugar content of monk fruit products The method for detecting the total sugar content of monk fruit products in this application employs the first method, HPLC-RID detection, as described in GB 5009.8-2016.

[0159] 2. Taste test of samples containing mogroside II and mogroside III Mogroside II and mogroside III are representative components of immature monk fruit and have a bitter taste, with mogroside II being particularly bitter (Chinese Academy of Sciences, Guangxi Institute of Botany, Research on Monk Fruit Cultivation and Chemistry, 1st edition, Nanning: Guangxi Science and Technology Press, 2010.5). If the bitter components in the product cannot be controlled to a certain level, it will directly affect the overall taste of the product. Samples obtained after separation and elution using resin column A in Example 1 were collected, purified, and then dissolved in pure water at concentrations ranging from low to high. The taste was then recorded. The results are shown in Table 25 below.

[0160] [Table 25]

[0161] The test results in Table 25 above support the literature's description that mogroside II and mogroside III are bitter components. It was found that products containing mogroside II and mogroside III at lower formulation concentrations still exhibit a significant bitter taste, explaining that the content of these two components in the product must be controlled before use.

[0162] 3. Comparative study of the sweetness characteristics of 3.11-oxomogroside V (11-O-mogroside V) and mogroside V. Samples were collected, separated, and prepared to obtain a sample with a purity of approximately 77.3% for 11-oxomogloside V. An appropriate amount was taken and prepared to achieve a relative sucrose sweetness of 5%. Additionally, an appropriate amount of an unseparated and unpurified conventional sample (ion-exchanged but not multi-column separated and purified) was taken and prepared to achieve a relative sucrose sweetness of 5%. Taste tests were conducted and the results were compared to determine and recorded the peak time of sweetness and the duration of sweetness. The results are shown in Table 26.

[0163] [Table 26]

[0164] The 11-oxomogroside V contained in mogroside products is beneficial in improving the speed of the sweetness response and shortening the after-sweetness performance to some extent, bringing the overall taste of the sweetener closer to sucrose and improving the texture of the product. Obtaining high-purity mogroside V does not mean that the higher the purity, the better. A better combination and process solution is to remove components that negatively affect the flavor of the product, such as mogroside II and mogroside III, while retaining components that can enhance the overall flavor, such as 11-oxomogroside V, and maintaining a certain content level.

[0165] The foregoing describes only preferred embodiments of this application and is not intended to limit this application to other forms. Those skilled in the art can use the above-disclosed technical content to make equivalent modifications or changes to equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of this application without departing from the content of the proposed technical concept of this application remain within the scope of protection of the proposed technical concept of this application.

Claims

1. Based on the total weight of the sweetening composition, 70% to 85% mogroside V, 5% to 12% 11-O-mogroside V, 0-0.5% siamenoside I and 0-0.05% mogroside II and mogroside III, 0-0.5% protein and 0-0.5% total sugar A sweetening composition characterized by containing [a certain ingredient].

2. The extraction step involves extracting monk fruit to obtain an extract; A clarification step of clarifying the extract to obtain a clarified liquid; An ion exchange treatment step is performed by sequentially treating the clarified liquid with a strongly acidic cation exchange resin column and a weakly basic anion exchange resin column to obtain an ion exchange treatment solution; The ion exchange treatment solution is placed in an adsorption resin column combination and subjected to multi-stage adsorption separation to eluate, the eluate is collected and dried to obtain a sweet composition, and the process is a multi-stage adsorption separation step. Includes, A method for preparing a sweetening composition, characterized in that the ion exchange treatment step is performed before the multi-stage adsorption separation step.

3. The ion exchange treatment solution is sequentially introduced into the 1st, 2nd, ..., nth (n≧2) adsorption resin columns of the adsorption resin column combination to perform multi-stage adsorption separation, and the separated column eluent obtained after passing through each adsorption resin column directly enters the next adsorption column, or The preparation method according to claim 2, characterized in that the ion exchange treatment solution is sequentially introduced into the 1st, 2nd, ..., nth (n≧2) adsorption resin columns of the adsorption resin column combination to perform multi-stage adsorption separation, after passing through each of the adsorption resin columns from the 1st to the (x-1)th (n≧x≧2)th adsorption resin columns, the separated column eluates obtained after passing through each adsorption resin column are collected and combined, and then charged into the next adsorption resin column for separation, and the column eluates adsorbed and collected in the (x-1)th adsorption resin column are sequentially introduced into the xth, x+1st, x+2nd, ..., nth adsorption resin columns for separation.

4. The preparation method according to claim 3, characterized in that, after multi-stage adsorption separation, the sweet composition of this application is obtained from each of the adsorption resin columns from the xth to nth (2 ≤ x ≤ n) adsorption resin columns.

5. The preparation method according to claim 4, characterized in that, in the multi-stage adsorption separation step, the mass of mogroside V in the ion exchange treatment solution that has passed through the 1st, ..., x-1 (2 ≤ x ≤ n) adsorption resin columns is 1 to 4 times, preferably 1 to 3 times, more preferably 1.5 to 3 times, the total adsorbed mass of the 1st to x-1 adsorption resin columns when mogroside V is saturated adsorbed.

6. The preparation method according to any one of claims 2 to 5, characterized in that, after the clarification step and before the ion exchange treatment step, a mogroside concentration step using an adsorption resin column A'' is also included.

7. The adsorption resin column A'' is a weakly polar macroporous adsorption resin column, Preferably, the adsorption resin column A'' is a macroporous adsorption resin column whose backbone material is polyethylenedivinylbenzene. Preferably, the particle size of the resin is 250 μm to 1250 μm, preferably 250 μm to 450 μm. The preparation method according to claim 6, preferably characterized in that the resin model of the adsorption resin column A'' is selected from LX-15, D101, AB-8, SP700, XD16N, and XD1600N, and preferably SP700.

8. The preparation method according to any one of claims 2 to 7, characterized in that the extract is obtained by hot water extraction.

9. The preparation method according to any one of claims 2 to 8, characterized in that the clarified liquid is obtained by centrifugation and / or filtration.

10. The particle size of the resin in the aforementioned strongly acidic cation exchange resin column is 300 to 1100 μm, preferably 300 to 600 μm. Preferably, the resin of the strongly acidic cation exchange resin column is one selected from Marathon MSC, AMBERLITE FPC22H, DIAION SK series and DIAION PK series, and preferably Marathon MSC. Preferably, the particle size of the resin in the weakly basic anion exchange resin column is 350 to 1250 μm, and more preferably 350 to 600 μm. The preparation method according to any one of claims 2 to 9, preferably characterized in that the resin of the weakly basic anion exchange resin column is one selected from Marathon WBA, SCAV2, FPA53 and LX-T5, and preferably Marathon WBA or SCAV2.

11. Each adsorption resin column used in the multi-stage adsorption separation step is a weakly polar macroporous adsorption resin column. Preferably, each of the adsorption resin columns is a macroporous adsorption resin column whose backbone material is polyethylenedivinylbenzene. The preparation method according to any one of claims 2 to 10, preferably characterized in that the resin of each adsorption resin column is one selected from LX-15, D101, AB-8, SP700, XD16N, and XD1600N, and preferably SP700.

12. The preparation method according to any one of claims 2 to 11, characterized in that the multi-stage adsorption separation is performed using two or more, preferably two to six, adsorption resin columns.

13. Based on the total weight of the sweetening composition, the sweetening composition is 70% to 85% mogroside V, 5% to 12% 11-O-mogroside V, 0-0.5% siamenoside I and 0-0.05% mogroside II and mogroside III, 0-0.5% protein and 0-0.5% total sugar A preparation method according to any one of claims 2 to 12, characterized by including the following:

14. In the multi-stage adsorption separation step, after the preparation of the ion exchange treatment solution is complete, the adsorption resin column combination is first eluted with water or organic solvent 1, and then eluted with organic solvent 2. Preferably, the organic solvent 1 is ethanol with a concentration of 10% to 30%. Preferably, the organic solvent 2 is one or two selected from hydrophilic ketones and low-hydride alcohols. Preferably, the organic solvent 2 is one or more selected from methanol, ethanol, propanol, and acetone. Preferably, the organic solvent 2 is ethanol with a concentration of 55% or more, The preparation method according to any one of claims 2 to 13.

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