Sweetness composition and preparation method thereof

Abstract

A sweetness composition, based on the total weight of the sweetness composition, comprising: 70% to 85% of mogroside V; 5% to 12% of 11-O-mogroside V; 0 to 0.5% of simonoside I; 0 to 0.05% of mogroside II and mogroside III; 0 to 0.5% of protein; and 0 to 0.5% of total sugar. Also disclosed is a method for preparing the sweetness composition, comprising the following steps: extracting Siraitia grosvenorii to obtain an extract; clarifying the extract to obtain a clarified liquid; sequentially treating the clarified liquid with a strong acid cation exchange resin column and a weak alkaline anion exchange resin column to obtain an ion exchange treated liquid; and the ion exchange treated liquid entering an adsorption resin column combination for multi-stage adsorption separation and elution, collecting the eluate, and drying to obtain the sweetness composition, wherein, the ion exchange treatment step needs to be performed before the multi-stage adsorption separation step.

Description

Sweet composition and preparation method thereof Technical Field

[0001] The present application relates to the field of extraction technology, and in particular to a sweetening composition and a preparation method thereof. Background Art

[0002] Monk fruit (Shengguo) is the mature fruit of a perennial herbaceous vine in the Cucurbitaceae family. Its glycosides are cooling in nature and enter the lung and large intestine meridians, clearing heat and moistening the lungs, while also promoting bowel movements. It is a commonly used Traditional Chinese Medicine in my country and one of the first medicinal and edible plants approved by the Ministry of Health. Monk fruit is primarily produced in Guilin, Guangxi, with large-scale cultivation in Yongfu, Lingui, and Longsheng counties in Guangxi. In recent years, with the rapid growth of demand for mogroside sweeteners, its cultivation has gradually expanded to cities and counties in Guizhou and Hunan, neighboring Guilin.

[0003] Mogrosides are a collective term for the sweet-tasting cucurbitane triterpenoid saponins found in Luo Han fruit. They are a class of triterpene glucosides whose aglycones are triterpenoid alcohols. Mogroside V is the primary sweetening component. It is non-toxic, low in calories, highly sweet, and thermally stable, making it a key indicator for content control in the National Food Safety Standard for Mogrosides (GB1886.77-2016), a natural sweetener. Currently, mogrosides are 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 primarily 40-50%, with only a few reaching 60% purity. In addition to the various extremely sweet glycosides, these mogroside products also contain small amounts of bitter glycosides such as mogroside II and mogroside III. This is due to the bitterness introduced by some immature or less mature monk fruit when the fresh monk fruit is extracted, which negatively affects the overall taste of the mogroside products. Furthermore, current mogroside products typically still retain a high protein content and a certain sugar content derived from fresh monk fruit. The presence of these nutritional ingredients imposes certain limitations on the product's shelf life and applications requiring high energy intake.

[0004] Summary of the Invention

[0005] Although researchers have also developed some methods for preparing high-purity mogroside V to improve the quality of mogroside products, such as patents CN110669095A "A method for extracting high-purity mogroside V" and CN101402665B "A method for preparing mogroside V", although both obtain mogroside V products with a purity of more than 99%, their processes all use a purification technology based on reverse-phase chromatography with C18 filler with a particle size of 0.45μm. The technology uses medium- and high-pressure preparative liquid chromatography, with large equipment investment and small mass production scale. In addition, toxic solvents such as acetonitrile are used in the process, which is not conducive to food safety control.

[0006] Patent CN101407535B discloses a method for preparing high-purity mogroside V, which involves dissolving and filtering with methanol, adding acetone to the filtrate for precipitation, subjecting the filtered precipitate to silica gel chromatography, eluting with a mixed solvent of fatty acid esters and ethanol, and concentrating and crystallizing the eluate to obtain mogroside V with a purity exceeding 98%. This process uses organic solvents such as acetone, fatty acid esters, and ethanol, and the silica gel chromatography process produces a small amount of product, making reuse difficult and prohibiting large-scale production.

[0007] EP2622969A1 discloses a sweetener mixture comprising mogroside V with a purity of 60-70%. The high-purity mogroside can be obtained by a method for preparing a highly purified mogroside mixture from a low-purity mogroside mixture. Although the method innovatively uses a multi-column adsorption column system, a complex combination of acidic, alkaline and aqueous alcohol solutions is used to elute impurities. In addition, the content of mogroside V obtained by this method is relatively low, and the method fails to provide a method and effect for controlling the content of bitter glycosides derived from immature or under-mature monk fruit raw materials, namely mogroside II and mogroside III. It also fails to provide a method and effect for controlling the content of protein and sugar that affect product application.

[0008] CN104059122A discloses a method for preparing high-purity mogroside V. The core of the method for purifying mogroside V is the innovative use of unconventional low-particle-size 100-200 mesh (74-149 μm) anion and cation exchange resins as chromatographic fillers, and a specific pH gradient aqueous solution for elution to obtain the target product. This is an atypical innovative method for improving mogroside V using ion exchange resins but not for the purpose of deionization. The method also fails to provide a method and effect for controlling the content of bitter glycosides derived from immature or under-mature mogroside raw materials, namely mogroside II and mogroside III, and also fails to provide a method and effect for controlling the content of protein and sugar that affect product application.

[0009] In order to overcome the defects of the prior art, the present application provides a low-protein, low-bitter glycoside, low-sugar content, high-purity sweet glycoside composition of mogroside V and a preparation method thereof.

[0010] The specific technical solutions of this application are as follows:

[0011] In one aspect, the present application provides a sweetening composition, wherein, based on the total weight of the sweetening composition, it comprises:

[0012] 70% to 85% mogroside V;

[0013] 5% to 12% 11-O-mogroside V;

[0014] 0-0.5% simenoside I;

[0015] 0-0.05% mogroside II and mogroside III;

[0016] 0-0.5% protein and 0-0.5% total sugars.

[0017] On the other hand, the present application also provides a method for preparing a sweet composition, which comprises the following steps:

[0018] Extraction: extracting Momordica grosvenori to obtain an extract;

[0019] Clarifying: clarifying the extract to obtain a clarified solution;

[0020] Ion exchange treatment: the clarified liquid is sequentially treated with a strong acid cation exchange resin column and a weak base anion exchange resin column to obtain an ion exchange treated liquid;

[0021] Multi-stage adsorption separation: the ion exchange treated liquid enters the adsorption resin column combination for multi-stage adsorption separation, elution, collection of the eluate, and drying to obtain a sweet composition;

[0022] Among them, the ion exchange treatment step needs to be carried out before the multi-stage adsorption separation step.

[0023] Furthermore, in the method for preparing the sweet composition of the present application, the ion exchange treated liquid sequentially enters the first, second, ..., nth (n≥2) adsorption resin columns of the adsorption resin column combination for multi-stage adsorption separation, wherein the separated column liquid obtained by passing through each adsorption resin column directly enters the subsequent adsorption column; or

[0024] The ion exchange treatment liquid is sequentially passed through the 1st, 2nd, ..., nth (n≥2) adsorption resin columns of the adsorption resin column combination for multi-stage adsorption separation, wherein, after passing through each of the 1st to (x-1) (n≥x≥2) adsorption resin columns, the separated column liquid obtained after passing through each adsorption resin column is collected and combined, and then fed to the next adsorption resin column for separation; and then the column liquid collected after adsorption by the (x-1)th adsorption resin column enters the xth, x+1th, x+2th, ..., nth adsorption resin columns in sequence for adsorption separation.

[0025] Furthermore, in the method for preparing the sweet composition of the present application, after multi-stage adsorption separation, the sweet composition of the present application is obtained from each of the xth to nth (2≤x≤n) adsorption resin columns.

[0026] Furthermore, in the preparation method of the sweet composition of the present application, in the multi-stage adsorption separation step, the mass of mogroside V in the ion exchange treatment liquid passing through the 1st, ..., x-1th (2≤x≤n) adsorption resin columns should be 1 to 4 times the sum of the adsorption masses of mogroside V when saturated adsorbed by the 1st to x-1th adsorption resin columns, preferably 1 to 3 times, and more preferably 1.5 to 3 times.

[0027] Furthermore, in the preparation method of the sweet composition of the present application, after the clarification step and before the ion exchange treatment step, a mogroside enrichment step using an adsorption resin column A" is further included.

[0028] Furthermore, in the preparation method of the sweet composition of the present application, the adsorption resin column A" is a weakly polar macroporous adsorption resin column;

[0029] Preferably, the adsorption resin column A" is a macroporous adsorption resin column whose skeleton material is polyethylene divinylbenzene;

[0030] Preferably, the particle size of the resin is 250 μm to 1250 μm, preferably 250 μm to 450 μm;

[0031] Preferably, the resin model of the adsorption resin column A" is selected from one of LX-15, D101, AB-8, SP700, XD16N and XD1600N, preferably SP700.

[0032] Furthermore, in the preparation method of the sweet composition of the present application, hot water is used to extract to obtain the extract.

[0033] Furthermore, in the method for preparing the sweet composition of the present application, the clarified liquid is obtained by centrifugation and / or filtration.

[0034] Furthermore, in the method for preparing the sweet composition of the present application, the particle size of the resin of the strong acid cation exchange resin column is 300 to 1100 μm, preferably 300 to 600 μm;

[0035] Preferably, the resin of the strong acid cation exchange resin column is selected from one of Marathon MSC, AMBERLITE FPC22H, DIAION SK series and DIAION PK series, preferably Marathon MSC;

[0036] 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;

[0037] Preferably, the resin of the weakly basic anion exchange resin column is selected from one of Marathon WBA, SCAV2, FPA53 and LX-T5, preferably Marathon WBA or SCAV2.

[0038] 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;

[0039] Preferably, each of the adsorption resin columns is a macroporous adsorption resin column whose skeleton material is polyethylene divinylbenzene;

[0040] Preferably, the resin of each adsorption resin column is selected from one of LX-15, D101, AB-8, SP700, XD16N and XD1600N, preferably SP700.

[0041] Furthermore, in the method for preparing the sweet composition of the present application, the multi-stage adsorption separation is a multi-stage adsorption separation through two or more, preferably two to six, adsorption resin columns.

[0042] Furthermore, in the preparation method of the sweet composition of the present application, the sweet composition comprises, based on the total weight of the sweet composition:

[0043] 70% to 85% mogroside V;

[0044] 5% to 12% 11-O-mogroside V;

[0045] 0-0.5% simenoside I;

[0046] 0-0.05% mogroside II and mogroside III;

[0047] 0-0.5% protein and 0-0.5% carbohydrates.

[0048] Furthermore, in the method for preparing the sweet composition of the present application, in the multi-stage adsorption separation step, after the ion exchange treatment liquid is fed, the adsorption resin column combination is first eluted with water or organic solvent 1, and then eluted with organic solvent 2;

[0049] Preferably, the organic solvent 1 is ethanol with a concentration of 10% to 30%;

[0050] Preferably, the second organic solvent is selected from one or both of hydrophilic ketones and low-alcohols;

[0051] Preferably, the second organic solvent is selected from one or more of methanol, ethanol, propanol and acetone;

[0052] Preferably, the second organic solvent is ethanol with a concentration of 55% or more.

[0053] Effects of the Invention

[0054] The method adopted in the present application first removes ionic substances such as inorganic salts, organic acids, alkaloids, flavonoids, and proteins in the monk fruit extract with a combination of cation and anion exchange resins, and then forms a molecular exclusion chromatography for the adsorption of mogroside components by multi-column adsorption resin chromatography. By combining the amount of resin in each column and controlling the feed amount, the slight difference in the binding ability of each mogroside component with the resin is used to accumulate the bitter mogroside II and mogroside III in the resin that first enters the adsorption resin column, and pushes the components with slightly weaker binding ability to the resin, such as 11-O-mogroside V and mogroside V, to the resin columns connected in series later, and then eluting with different concentrations of alcohol to obtain the target product. In addition to separating and removing the bitter glycosides, the present application also forms a better combination of 11-O-mogroside V and mogroside V, so that the product has a purer mogroside V and a better taste. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] FIG1 is an HPLC spectrum of various glycoside components in Momordica grosvenori according to a specific embodiment of the present application.

[0056] FIG2 is a high performance liquid chromatogram of the product obtained by eluting with 60% alcohol from column D in Example 5 of the present application.

[0057] FIG3 is a high performance liquid chromatogram of the product obtained by eluting with 60% alcohol from column E in Example 5 of the present application. DETAILED DESCRIPTION

[0058] Below, the exemplary embodiments of the present application are described, including various details of the embodiments of the present application to facilitate understanding, which should be considered as merely exemplary. Therefore, it should be appreciated by those skilled in the art that various changes and modifications may be made to the embodiments described herein without departing from the scope and spirit of the present application. Similarly, for clarity and conciseness, the description of known functions, structures, and reagents has been omitted in the following description.

[0059] In one aspect, the present application provides a sweetening composition, wherein, based on the total weight of the sweetening composition, it comprises:

[0060] 70% to 85% mogroside V;

[0061] 5% to 12% 11-O-mogroside V;

[0062] 0-0.5% simenoside I;

[0063] 0-0.05% mogroside II and mogroside III;

[0064] 0-0.5% protein and 0-0.5% total sugars.

[0065] The mogroside V of the present application is a cucurbitane-type triterpenoid saponin, and its chemical structure is known in the art. In some embodiments, the sweetening composition may include, 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 therebetween of mogroside V, based on the total weight of the sweetening composition.

[0066] The 11-O-mogroside V of the present application is a cucurbitane-type triterpenoid saponin and a sweetener. Its chemical structure is known in the art. It has strong antioxidant activity and has a significant inhibitory effect on reactive oxygen species. In some embodiments, the sweetening composition may include, 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 therebetween, based on the total weight of the sweetening composition.

[0067] Simenoside I is a cucurbitane-type triterpenoid saponin and the sweetest component of mogroside. Its sweetness is about 460 to 563 times that of sucrose, but its natural abundance in monk fruit is extremely low. In some embodiments, based on the total weight of the sweetening composition, the sweetening composition may include, 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%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.50%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.59%, 0.60%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.70%, %, 0.24%, 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 therebetween of simenoside I.

[0068] Mogrosides typically have varying numbers (2 to 6) of glucose units attached to carbon 3 and carbon 24 of the triterpene backbone. Mogroside II is the simplest mogroside, having one glucose residue attached to carbon 3 and one glucose residue attached to carbon 24. Mogroside III differs in that it has an additional glucose residue attached to carbon 24. In some embodiments, the sweetening composition may include mogroside II and mogroside III in an amount, for example, in an amount of 0%, 0.001%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, or any range therebetween, by weight of the total weight of the sweetening composition.

[0069] In some embodiments, the sweetening composition may comprise, 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 thereof, based on the total weight of the sweetening composition. The protein content of the present application can be determined by any suitable method or technique known in the art, for example, the first method of GB5009.5-2016 can be used for detection.

[0070] In some embodiments, the sweetening composition may include, 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 thereof, based on the total weight of the sweetening composition. The total sugar content of the present application refers to the sum of the contents of monosaccharides and disaccharides based on dry weight. The total sugar content of the present application can be determined by any suitable method or technique known in the art, for example, the first method of GB 5009.8-2016, HPLC-RID detection can be used.

[0071] On the other hand, the present application also provides a method for preparing a sweet composition, which comprises the following steps:

[0072] Extraction: extracting Momordica grosvenori to obtain an extract;

[0073] Clarifying: clarifying the extract to obtain a clarified solution;

[0074] Ion exchange treatment: the clarified liquid is sequentially treated with a strong acid cation exchange resin column and a weak base anion exchange resin column to obtain an ion exchange treated liquid;

[0075] Multi-stage adsorption separation: the ion exchange treated liquid enters the adsorption resin column combination for multi-stage adsorption separation, elution, collection of the eluate, and drying to obtain a sweet composition;

[0076] Among them, the ion exchange treatment step needs to be carried out before the multi-stage adsorption separation step.

[0077] This application, based on the conventional production process of mogrosides: a combined process of monk fruit crushing, water extraction, macroporous resin adsorption, and resin decolorization, studies the selectivity differences in the adsorption of mogroside V, other mogroside components, and other components in monk fruit on macroporous adsorption resins. It was found that when the monk fruit extract treated with ion exchange is passed through a combination of macroporous resin columns, the distribution of mogroside V, other mogroside components, and other components in each column can be adjusted by controlling the feed rate. Sweet compositions containing different combinations of mogrosides can be obtained by elution and drying from each column, wherein the content of mogroside V in the sweet compositions can reach 70% to 85%.

[0078] In some embodiments, the preparation method of the sweet composition of the present application includes: contacting Luo Han Guo with water to prepare an extract, wherein the extract contains mogroside V, other terpene glycosides and sugars; clarifying the extract to remove pectin and protein, etc., so as to facilitate subsequent processing; contacting the clarified liquid obtained after clarification with a strong acid cation exchange resin column and a weak base anion exchange resin column in sequence for decolorization, deacidification, and further deproteinization; contacting the ion exchange treated liquid obtained after the ion exchange treatment with an adsorption resin column combination for elution, so that the content of mogroside V, 11-O-mogroside V and simenoside I in the eluate is maximized, while the content of mogroside II, mogroside III and protein is minimized, and finally drying is performed to obtain the sweet composition of the present application.

[0079] In some embodiments, the extract is obtained by extraction with hot water, and the temperature of the hot water can 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.

[0080] In some embodiments, the extraction step comprises: crushing the fresh or dried monk fruit and extracting with hot water to obtain the extract.

[0081] In the present application, clarification can be performed 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 performed by phosphoric acid or pectinase treatment. Pectinase can be used in the form of a commercially available enzyme mixture containing pectinase to cleave pectin from the extract and precipitate pectin-stabilized peptides and proteins.

[0082] In some embodiments, the extract is initially clarified by centrifugation, and the clear liquid is further clarified by ultrafiltration through a ceramic membrane to obtain a clarified liquid.

[0083] The method of the present application utilizes ion exchange resins to remove inorganic salts, proteins, amino acids, and charged components from the clarified liquid obtained from Momordica grosvenori, while maximally retaining the amount of certain terpene glycosides, particularly mogroside V, which helps to obtain a sweet composition with a pure taste. Ion exchange resins are polymers that can exchange specific ions within the polymer with ions in a solution passing through the resin. Ion exchange resins can be insoluble in acids or bases, and their salts are also insoluble, allowing the resin to exchange with positively charged ions (cation exchange resins) or negatively charged ions (anion exchange resins). The present application sequentially uses a strong acid cation exchange resin column and a weak base anion exchange resin column for ion exchange treatment, which can retain the strong sweetness in the sweet composition to the greatest extent. 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.

[0084] In some embodiments, the compound responsible for a grassy or earthy taste or odor and a bitter taste 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 a bitter phenol. 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 polyphenol. In some embodiments, the compound is a bitter terpene glycoside.

[0085] In some embodiments, the particle size of the resin of the strong acid cation exchange resin column is 300 to 1100 μm, and the particle size of the resin of the weak base anion exchange resin column is 350 to 1250 μm.

[0086] In some embodiments, the particle size of the resin of the strong acid cation exchange resin column is 300-600 μm, and the particle size of the resin of the weak base anion exchange resin column is 350-600 μm.

[0087] In some embodiments, the resin of the strong acid cation exchange resin column is selected from one of Marathon MSC, AMBERLITE FPC22H, DIAION SK series and DIAION PK series, and the resin of the weak base anion exchange resin column is selected from one of Marathon WBA, SCAV2, FPA53 and LX-T5.

[0088] In some embodiments, the resin of the strong acid cation exchange resin column is Marathon MSC, and the resin of the weak base anion exchange resin column is Marathon WBA or SCAV2.

[0089] Typically, based on the selectivity of macroporous adsorption resins, in typical aqueous and alcoholic solvent systems, mogroside IV, simenoside I, mogroside II, mogroside III, mogrool, and other non-sweet glycosides, which are less polar than mogroside V, are retained more strongly on the resin than mogroside V. Meanwhile, components with greater polarity than mogroside V, such as mogroside 11-O-mogroside V and other non-sweet glycosides, are less retained than mogroside V. These differences in resin selectivity are not readily apparent in typical single-column adsorption and are difficult to fully exploit. However, when using a multi-column system, components in the feed can be accumulated and amplified based on differences in polarity or adsorption selectivity, creating an effect similar to size exclusion chromatography. When loading the resin column, components with less polarity or relatively strong adsorption properties gradually accumulate in the columns ranked higher. With sufficient feed volume, these components will dominate the adsorption capacity of the leading resin columns, while mogroside V and other more polar components, due to less competitive adsorption capacity, are eluted or pushed to later columns. The components entering the rear-end column also have differences in polarity and resin adsorption capacity. Components with stronger polarity than mogroside V will be competitively adsorbed by mogroside V and pushed to the rear columns or the column liquid. This makes the composition of the components and the content of mogroside V in adsorption columns of different orders have significant differences.

[0090] In some embodiments, the ion exchange treatment liquid sequentially passes through the 1st, 2nd, ..., nth (n≥2) adsorption resin columns of the adsorption resin column combination for multi-stage adsorption separation, wherein, after passing through each of the 1st to (x-1) (n≥x≥2) adsorption resin columns, the separated column liquid obtained after passing through each adsorption resin column is collected and combined, and then fed to the next adsorption resin column for separation; then, the column liquid collected after adsorption by the (x-1)th adsorption resin column enters the xth, x+1th, x+2th, ..., nth adsorption resin columns in sequence for adsorption separation. In some embodiments, the ion exchange treatment liquid sequentially enters the 1st, 2nd, ..., nth (n≥2) adsorption resin columns of the adsorption resin column combination for multi-stage adsorption separation, wherein, the separated column liquid obtained after passing through each adsorption resin column is not collected but directly enters the subsequent adsorption column. During serial separation, the separated components are distributed sequentially within the resin layer based on differences in molecular binding strength to the resin (strongly adsorbed substances are distributed later, weakly adsorbed substances are discharged first). After collecting and merging the materials between columns, the order of the components separated by the first column is disrupted, making it difficult to increase the separation of the subsequent column. Therefore, of the two aforementioned embodiments, the latter has a superior adsorption separation effect compared to the former. In the method of the present application, after multi-stage adsorption separation, the sweet composition of the present application is obtained from each of the xth to nth (2≤x≤n)th adsorption resin columns.

[0091] In some embodiments, the multi-stage adsorption separation described herein is performed using two or more, preferably two to six, adsorption resin columns. For example, the multi-stage adsorption separation may be performed using two, three, four, five, or six adsorption resin columns, more preferably two, three, or four adsorption resin columns. In the present application, the resin types of the adsorption resin columns may be the same or different, and the resin amounts of the adsorption resin columns may be the same or different.

[0092] In some embodiments, in the multi-stage adsorption separation step, the mass of mogroside V in the ion exchange treatment liquid passing through the 1st, ..., x-1th (2≤x≤n) adsorption resin columns should be 1 to 4 times the sum of the adsorption masses of the 1st to x-1th adsorption resin columns when saturated with mogroside V. For example, it can be 1 time, 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2 times, 2.1 times, 2.2 times, 2.3 times, 2.4 times, 2.5 times, 2.6 times, 2.7 times, 2.8 times, 2.9 times, 3 times, 3.1 times, 3.2 times, 3.3 times, 3.4 times, 3.5 times, 3.6 times, 3.7 times, 3.8 times, 3.9 times, 4 times, etc., preferably 1 to 3 times, and more preferably 1.5 to 3 times. The multiple is relative to the volume of resin in the column, which cannot achieve high purity. Too large a multiple means that there are many strongly adsorbed impurities (weakly polar substances, mogroside II, mogroside III, etc.) and they exceed the adsorption and retention capacity of the resin in column x-1, and are pushed to the xth column, increasing the impurities in column x and affecting the purity or taste of the purified sweet glycosides. Too small a feed rate results in low utilization of the resin in column x-1, and too little sweet glycoside enters columns x-n. Although a higher purity product can be obtained, the yield is too low, and the economic benefits are very poor.

[0093] In some embodiments, after the clarification step and before the ion exchange treatment step, a mogroside enrichment step is further included using an adsorption resin column A". In some embodiments, the mogroside enrichment step includes: enriching mogroside by adsorption on the clarified liquid through 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 filtered supernatant for use in the subsequent ion exchange treatment step.

[0094] In some embodiments, in the enrichment step, the adsorption resin column A" is a macroporous adsorption resin column whose framework material is polyethylene divinylbenzene; and the particle size of the resin is 250 μm to 1250 μm.

[0095] In some embodiments, in the enrichment step, the resin model of the adsorption resin column A" includes but is not limited to commonly used macroporous adsorption resins such as LX-15, D101, AB-8, SP700, XD16N and XD1600N, preferably SP700.

[0096] In some embodiments, each adsorption resin column used in the multi-stage adsorption separation step is a weak polar macroporous adsorption resin column; each adsorption resin column is preferably a macroporous adsorption resin column whose skeleton material is polyethylene divinylbenzene; 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, preferably SP700.

[0097] In some embodiments, the sweetening composition can include, 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 therebetween of mogroside V, based on the total weight of the sweetening composition.

[0098] In some embodiments, the sweetening composition may include, 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 therebetween of 11-O-mogroside V, based on the total weight of the sweetening composition.

[0099] In some embodiments, the sweetening composition may comprise, 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. %, 0.24%, 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 therebetween of simenoside I.

[0100] In some embodiments, the sweetening composition may include mogroside II and mogroside III in an amount of, for example, 0%, 0.001%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05% or any range therebetween, of the total weight of the sweetening composition.

[0101] In some embodiments, the sweetening composition may comprise, 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% protein, or any range therebetween, based on the total weight of the sweetening composition.

[0102] In some embodiments, the sweetening composition may comprise, 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% total sugars, based on the total weight of the sweetening composition.

[0103] In some embodiments, in the multi-stage adsorption separation step, after the ion exchange treatment liquid is fed, 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.

[0104] In some embodiments, in the multi-stage adsorption separation step, after the ion exchange treatment liquid is fed, the adsorption resin column assembly is first eluted with 30% ethanol and then with 55% or more ethanol.

[0105] The sweetening compositions described herein can be used in food, beverage, pharmaceutical, or dietary supplement products.

[0106] Example

[0107] This application provides a general and / or specific description of the materials and experimental methods used in the experiments. In the following examples, the materials or instruments used without indicating the manufacturer are all conventional materials or instruments that can be obtained commercially.

[0108] Example 1

[0109] 300 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90°C was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0110] The ultrafiltration clear liquid was filled with 50L of macroporous resin (model SP700, particle size 250μm, specific surface area 1200m 2 / g) was treated with a chromatography column (diameter 30 cm * column height 90 cm) to adsorb mogroside, the column was washed with 200 L of purified water, and then eluted with 150 L of 60% alcohol. The alcohol eluate was collected and concentrated under reduced pressure to obtain a mogroside concentrate b after the alcohol was removed, with a net weight of 13.8 kg, a solid content of 30% in the concentrate, and a content of mogroside V in the concentrate as dry matter of 26.5%;

[0111] The concentrated solution is diluted with purified water to a solid content of 5% for ion exchange treatment;

[0112] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series. The strong-acid cation exchange resin is DOWEX Marathon MSC, with a particle size of 550-900 μm, and the weak-base anion exchange resin is DOWEX Marathon WBA, with a particle size of 475-575 μm. Each column is loaded with 15 L of resin. The diluted concentrate is sequentially fed into the cation and anion exchange resin columns. After the feed is complete, the columns are washed with purified water. The column flowthrough treated with the ion exchange resin and the wash water are combined to obtain a mixed deionized solution C with a solids content of 2.8%, a volume of 65.9 L, and a dry matter content of 59.6% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0113] The multi-stage adsorption separation column group consists of three resin columns each containing 6 L of macroporous adsorption resin (SP700), numbered A, B, and C in sequence, and all are regenerated and washed with water for use.

[0114] The above-mentioned deionized solution c was passed through macroporous adsorption resin columns A, B, and C in sequence at a flow rate of 3 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 2.6 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12 L of purified water, and then column A was directly washed with 60% ethanol, and column B and C were eluted with 12 L of 25% alcohol respectively. Finally, each column was eluted with 60% alcohol separately, and the eluent was collected separately, concentrated and dried respectively, and the content of mogroside V and other glycoside components was detected. The results are shown in Table 1.

[0115] Table 1

[0116] Note: The net weight and content of each part are calculated on a dry matter basis, and the same applies to the following expressions.

[0117] Example 2

[0118] 356 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain 1200 L of ultrafiltrate clear solution a, which was then subjected to ion exchange treatment.

[0119] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 120 L of resin. The diluted concentrate is sequentially fed into the cation and anion exchange resin columns. The strong-acid cation exchange resin is DOWEX Marathon MSC, with a particle size of 550-900 μm, and the weak-base anion exchange resin is DOWEX Marathon WBA, with a particle size of 475-575 μm. After the feed is complete, the columns are washed with purified water. The column flowthrough treated with the ion exchange resin and the wash water are combined to produce a mixed deionized solution C with a solids content of 1.92%, a volume of 1300 L, and a dry matter content of 5.4% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0120] The multi-stage adsorption separation column group consists of four resin columns each containing 5 L of macroporous adsorption resin. The resin model is SP700, with a particle size of 250 μm and a specific surface area of ​​1200 m 2 / g, columns are numbered A, B, C, D, and all are regenerated and washed with water for use.

[0121] The above-mentioned deionized solution c was passed through macroporous adsorption resin columns A, B, C, and D in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treatment solution after A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly washed with 60% ethanol. Columns B, C, and D were eluted with 12 L of 25% alcohol, respectively. Finally, each column was eluted with 10 L of 60% alcohol. The eluates were collected separately, concentrated and dried, and the content of mogroside V and other glycosides was detected. The results are shown in Table 2.

[0122] Table 2

[0123] Example 3

[0124] 370 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear solution a, which was then subjected to ion exchange treatment.

[0125] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 130 L of resin. The strong-acid cation exchange resin is AMBERLITE FPC3500, with a particle size of 450-650 μm, and the weak-base anion exchange resin is SCAV2, with a particle size of 600-800 μm. Ultrafiltration supernatant (a) is sequentially fed into the cation and anion exchange resin columns. After the ultrafiltration, the columns are washed with purified water. The column flow-through and the wash water are combined to produce a mixed deionized solution (c). This solution has a solids content of 1.7%, a volume of 1200 L, and a dry matter content of 5.5% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0126] The multi-stage adsorption separation column group consists of three resin columns, each filled with 8L of macroporous adsorption resin. The resin model is XAD16N, with a particle size of 560-710μm and a specific surface area of ​​800m 2 / g, and the preliminary test shows that the adsorption capacity of XAD16N per unit resin glycoside in saturated state is 6.3g / 100ml.

[0127] The resin columns are numbered A, B, and C in sequence, and all are regenerated and washed with purified water before use.

[0128] The above-mentioned deionized solution c was passed through macroporous adsorption resin columns A, B, and C in sequence at a flow rate of 2.4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 3 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 2.5 BV of purified water, and each column was eluted with 16 L of 25% alcohol and then eluted with 16 L of 60% alcohol. The eluates were collected separately, concentrated and dried, and the contents of mogroside V and other glycosides were detected. The results are shown in Table 3.

[0129] Table 3

[0130] Example 4

[0131] 264 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 900 L of hot water at 85° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear solution a.

[0132] The ion exchange resin column consists of a strong acid cation exchange resin column and a weak base anion exchange resin column connected in series, with each column holding 92 L of resin. The strong acid cation exchange resin is AMBERLITE FPC3500 with a particle size of 450-650 μm, and the weak base anion exchange resin is SCAV2 with a particle size of 600-800 μm. The diluted concentrate is sequentially fed into the cation exchange and anion exchange resin columns. After the feed is complete, the columns are washed with purified water. The column flowthrough treated with the ion exchange resin and the washing water are combined to obtain a mixed deionized solution C with a solids content of 1.8%, a volume of 12.0 L, and a mogroside V content of 5.5% on a dry matter basis. Multi-stage adsorption separation and purification is then performed.

[0133] The multi-stage adsorption separation column group consists of two resin columns filled with 6L and 10L macroporous adsorption resin (model XAD1600N, particle size 350-450μm, specific surface area 800m 2 / g resin columns, numbered A and B in sequence, all of which were regenerated and washed with water for use.

[0134] The above-mentioned deionized solution c was passed through the A and B macroporous adsorption resin columns in sequence at a flow rate of 1.8 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 2.7 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 3BV of purified water, and then eluted with 2BV of 25% alcohol, and then eluted with 2BV of 60% alcohol. 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.

[0135] Table 4

[0136] Example 5

[0137] 300 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90°C was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0138] The ultrafiltration clear liquid was filled with 45L of macroporous resin (model LX-15, particle size 315~1250μm, specific surface area 750m 2 / g) was treated with a chromatography column (diameter 30 cm * column height 90 cm) to adsorb mogroside, the column was washed with 200 L of purified water, and then eluted with 150 L of 60% alcohol. The alcohol eluate was collected and concentrated under reduced pressure to obtain a mogroside concentrate b after the alcohol was removed, with a net weight of 13.6 kg, a solid content of 35% in the concentrate, and a content of mogroside V in the concentrate as dry matter of 25.8%;

[0139] The concentrated solution is diluted with purified water to a solid content of 5% for ion exchange treatment;

[0140] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series. The strong-acid cation exchange resin is DOWEX Marathon MSC, with a particle size of 550-900 μm, and the weak-base anion exchange resin is SCAV2, with a particle size of 600-800 μm. Each column is loaded with 5.5 L of resin. The diluted concentrate is sequentially fed into the cation and anion exchange resin columns. After the feed is complete, the columns are washed with purified water. The column flowthrough treated with the ion exchange resin and the wash water are combined to produce a mixed deionized solution C with a solids content of 3.0%, a volume of 63.5 L, and a dry matter content of 58.0% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0141] The multi-stage adsorption separation column group consists of 5 adsorption resin columns numbered A, B, C, D, and E. Column A is loaded with 5.5 L of model SP700 resin, column B is loaded with 4.5 L of model XAD1600N resin, and columns C, D, and E are all loaded with 2.25 L of XAD1600N resin. All are regenerated with 5% sodium hydroxide solution and washed with water for use.

[0142] The above-mentioned deionized solution c was passed through A, B, C, D, and E macroporous adsorption resin columns in sequence at a flow rate of 1.2 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was 1.7 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 2BV of purified water, and then columns A and B were directly eluted with 60% ethanol, and columns C, D, and E were eluted with 5L of 25% alcohol, and then eluted separately with 5L of 60% alcohol. The eluates were collected separately, concentrated and dried, and the contents of mogroside V and other glycosides and protein and total sugar were detected. The results are shown in Table 5.

[0143] Table 5

[0144] Example 6

[0145] 330 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0146] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 115 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, with a particle size of 450-650 μm, and the weak-base anion exchange resin is SCAV2, with a particle size of 600-800 μm. Solution a is sequentially passed through the cation and anion exchange resin columns. After the feed is complete, the columns are washed with purified water. The column flowthrough after ion exchange resin treatment and the wash water are combined to produce a mixed deionized solution c with a solids content of 2.1%, a volume of 1212 L, and a mogroside V content of 5.5% on a dry basis. Multi-stage adsorption separation and purification is then performed.

[0147] The multi-stage adsorption separation column group consists of 4 resin columns each containing 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0148] The above-mentioned deionized solution c was passed through A, B, C, and D macroporous adsorption resin columns in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2BV of 25% alcohol and then with 10 L of 60% alcohol. 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 6.

[0149] Table 6

[0150] Example 7

[0151] 356 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0152] An ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each loaded with 125 L of resin. The strong-acid cation exchange resin is AMBERLITE FPC22H, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation exchange and anion exchange resin columns. After completion of the feed, the columns are washed with purified water. The column flow-through after ion exchange resin treatment and the wash water are combined to obtain a mixed deionized solution c. The solution has a solids content of 2.3%, a volume of 1210 L, and a dry matter content of 5.4% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0153] The multi-stage adsorption separation column group consists of 4 resin columns each containing 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0154] The above-mentioned deionized solution c was passed through A, B, C, and D macroporous adsorption resin columns in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2BV of 25% alcohol and then with 10 L of 60% alcohol. 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 7.

[0155] Table 7

[0156] Example 8

[0157] 360 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0158] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 126 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is LX-T5. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.1%, a volume of 1370 L, and a dry matter content of 5.3% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0159] The multi-stage adsorption separation column group consists of 4 resin columns each containing 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0160] The deionized solution c was passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange-treated liquid after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2 BV of 25% alcohol and then with 10 L of 60% alcohol. The eluates were collected separately, concentrated and dried, and the contents of mogroside V and other glycosides, sugar content, and protein content were tested. The results are shown in Table 8.

[0161] Table 8

[0162] Example 9

[0163] 361 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear solution a.

[0164] The ion exchange resin column consists of a strong acid cation exchange resin column and a weak base anion exchange resin column connected in series, with each column holding 130 L of resin. The strong acid cation exchange resin is AMBERLITE FPC22H, and the weak base anion exchange resin is FPA53. Solution a is sequentially fed into the cation exchange and anion exchange resin columns. After the feed is complete, the columns are washed with purified water. The column flowthrough after ion exchange resin treatment and the wash water are combined to obtain a mixed deionized solution c. The solution has a solids content of 2.2%, a volume of 1238 L, and a dry matter content of 5.6% mogroside V. Multi-stage adsorption separation and purification is to be performed.

[0165] The multi-stage adsorption separation column group consists of 4 resin columns each containing 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0166] The deionized solution c was passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange-treated solution after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2 BV of 25% alcohol and then with 10 L of 60% alcohol. The eluates were collected separately, concentrated and dried, and the contents of mogroside V and other glycosides, sugar content, and protein content were tested. The results are shown in Table 9.

[0167] Table 9

[0168] Example 10

[0169] 366 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90°C was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0170] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each with a resin loading of 130 L. The strong-acid cation exchange resin model is FPC88 UPS Na, and the weak-base anion exchange resin model is FPA66 UPS. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are washed with purified water. The column flowthrough after ion exchange resin treatment and the wash water are combined to obtain a mixed deionized solution c. Its solid content is 2.3%, the volume is 1247 L, and the dry matter content of the solution is 5.4% mogroside V. Multi-stage adsorption separation and purification is to be performed.

[0171] The multi-stage adsorption separation column group consists of 4 resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0172] The deionized solution c was passed through macroporous adsorption resin columns A, B, C, and D at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange-treated solution after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2 BV of 25% alcohol and then with 10 L of 60% alcohol. The eluates were collected separately, concentrated and dried, and the contents of mogroside V and other glycosides, sugar content, and protein content were tested. The results are shown in Table 10.

[0173] Table 10

[0174] Example 11

[0175] 110 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 300 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0176] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 39 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.3%, a volume of 320 L, and a dry matter content of 5.7% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0177] The multi-stage adsorption separation column group consists of two resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A and B in sequence, and both are regenerated and washed with water for use.

[0178] The above-mentioned deionized solution c was passed through the A and B macroporous adsorption resin columns in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 1.2 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. The two columns were washed with 3BV of purified water, and then column A was directly eluted with 60% alcohol. The other columns were first eluted with 2BV of 25% alcohol, and then eluted with 2BV of 60% alcohol. 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.

[0179] Table 11

[0180] Example 12

[0181] 135 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 300 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear solution a.

[0182] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 47 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.5%, a volume of 396 L, and a dry matter content of 5.3% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0183] The multi-stage adsorption separation column group consists of two resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A and B in sequence, and both are regenerated and washed with water for use.

[0184] The above-mentioned deionized solution c was passed through the A and B macroporous adsorption resin columns in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 1.5 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. The two columns were washed with 3BV of purified water, and then column A was directly eluted with 60% alcohol. The other columns were first eluted with 2BV of 25% alcohol, and then eluted with 2BV of 60% alcohol. 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.

[0185] Table 12

[0186] Example 13

[0187] 200 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 600 L of hot water at 90°C was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0188] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 70 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.5%, a volume of 625 L, and a dry matter content of 5.3% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0189] The multi-stage adsorption separation column group consists of three resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A, B, and C in sequence, and all are regenerated and washed with water for use.

[0190] The above-mentioned deionized solution c was passed through macroporous adsorption resin columns A, B, and C in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 2.2 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 3BV of purified water, and then column A was directly eluted with 60% alcohol. The other columns were first eluted with 2BV of 25% alcohol, and then eluted with 2BV of 60% alcohol. 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.

[0191] Table 13

[0192] Example 14

[0193] 270 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 600 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear solution a.

[0194] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 95 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.4%, a volume of 754 L, and a dry matter content of 5.8% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0195] The multi-stage adsorption separation column group consists of three resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A, B, and C in sequence, and all are regenerated and washed with water for use.

[0196] The above-mentioned deionized solution c was passed through macroporous adsorption resin columns A, B, and C in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after column A was 3 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 3BV of purified water, and then column A was directly eluted with 60% alcohol. The other columns were first eluted with 2BV of 25% alcohol, and then eluted with 2BV of 60% alcohol. 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.

[0197] Table 14

[0198] Example 15

[0199] 565 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 900 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0200] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 200 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c. Its solids content is 2.35%, its volume is 1820 L, and its dry matter content of mogroside V is 5.73%. Multi-stage adsorption separation and purification is then performed.

[0201] The multi-stage adsorption separation column group consists of 4 resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0202] The above-mentioned deionized solution c was passed through A, B, C, and D macroporous adsorption resin columns in sequence at a flow rate of 5 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was 3.5 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 3BV of purified water, and then column A was directly eluted with 60% alcohol. The other columns were first eluted with 2BV of 25% alcohol, and then eluted with 2BV of 60% alcohol. 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.

[0203] Table 15

[0204] Example 16

[0205] 282 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear solution a.

[0206] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 100 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.1%, a volume of 918 L, and a dry matter content of 5.5% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0207] The multi-stage adsorption separation column group consists of 4 resin columns each filled with 5 L of macroporous adsorption resin (model AB-8), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0208] The above-mentioned deionized solution c was passed through A, B, C, and D macroporous adsorption resin columns in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2BV of 25% alcohol and then with 10 L of 60% alcohol. 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.

[0209] Table 16

[0210] Example 17

[0211] 263 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0212] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 92 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are rinsed with purified water. The column flow-through and the rinse water are combined to produce a mixed deionized solution c with a solids content of 2.1%, a volume of 866 L, and a dry matter content of 5.5% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0213] The multi-stage adsorption separation column group consists of 4 resin columns each filled with 5 L of macroporous adsorption resin (model D101), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0214] The above-mentioned deionized solution c was passed through A, B, C, and D macroporous adsorption resin columns in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12.5 L of purified water, and then column A was directly eluted with 60% alcohol. Columns B, C, and D were first eluted with 2BV of 25% alcohol and then with 10 L of 60% alcohol. 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.

[0215] Table 17

[0216] Example 18

[0217] 360 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 900 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0218] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 126 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are washed with purified water. The column flow-through and the wash water are combined to produce a mixed deionized solution c with a solids content of 2.4%, a volume of 1005 L, and a dry matter content of 5.8% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0219] The multi-stage adsorption separation column group consists of 5 resin columns each filled with 4 L of macroporous adsorption resin (model SP700), numbered A, B, C, D, and E in sequence, and all are regenerated and washed with water for use.

[0220] The above-mentioned deionized solution c was passed through macroporous adsorption resin columns A, B, C, D, and E in sequence at a flow rate of 4 L / h. After the feeding was completed, the total amount of mogroside V in the ion exchange treated liquid after columns A and B was 2.5 times the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12 L of purified water, and then column A was directly eluted with 10 L of 60% alcohol. Columns B, C, D, and E were first eluted with 2BV of 25% alcohol and then with 10 L of 60% alcohol. 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 18.

[0221] Table 18

[0222] Example 19

[0223] 360 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 900 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0224] The ion exchange resin column consists of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, each column holding 126 L of resin. The strong-acid cation exchange resin is DOWEX Marathon MSC, and the weak-base anion exchange resin is DOWEX Marathon WBA. Solution a is sequentially fed into the cation and anion exchange resin columns. After completion of the feed, the columns are washed with purified water. The column flow-through and the wash water are combined to produce a mixed deionized solution c with a solids content of 2.4%, a volume of 1005 L, and a dry matter content of 5.8% mogroside V. Multi-stage adsorption separation and purification is then performed.

[0225] The multi-stage adsorption separation column group consists of 4 resin columns each filled with 5 L of macroporous adsorption resin (model SP700), numbered A, B, C, and D in sequence, and all are regenerated and washed with water for use.

[0226] The above-mentioned deionized solution c was first passed through column A at a flow rate of 5 L / h, and the column liquid was collected. It was then passed through column B at a flow rate of 5 L / h, and the column liquid was collected. The column liquid was then passed through macroporous adsorption resin columns C and D in sequence. After the feeding was completed, the total amount of glycoside V in the ion exchange treated liquid after columns A and B was twice the saturated adsorption capacity of the corresponding amount of resin for mogroside V. Each column was washed with 12 L of purified water, and then column A was directly eluted with 10 L of 60% alcohol. Columns B, C, and D were first eluted with 2BV of 25% alcohol, and then eluted with 10 L of 60% alcohol. 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.

[0227] Table 19

[0228] Comparative Example 1

[0229] 300 kg of fresh monk fruit was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90°C was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain ultrafiltrate a.

[0230] The ultrafiltration clear liquid was filled with 50L of macroporous resin (model SP700, particle size 250μm, specific surface area 1200m 2 / g) was treated with a chromatography column (diameter 30 cm * column height 90 cm) to adsorb mogroside, the column was washed with 200 L of purified water, and then eluted with 150 L of 60% alcohol. The alcohol eluate was collected and concentrated under reduced pressure to obtain a mogroside concentrate b after the alcohol was removed, with a net weight of 13.8 kg, a solid content of 30% in the concentrate, and a content of mogroside V in the concentrate as dry matter of 26.5%;

[0231] The concentrated liquid is diluted with purified water to a solid content of 5% and then subjected to multi-stage adsorption separation treatment;

[0232] The multi-stage adsorption separation column set consisted of three resin columns, each loaded with 6 L of macroporous adsorption resin (SP700). The columns were sequentially numbered A, B, and C, and all were regenerated and washed with water before use. The ultrafiltrate supernatant a was passed through the A, B, and C macroporous adsorption resin columns at a flow rate of 3000 ml / h. After the feeding was completed, each column was washed with 12 L of purified water. Column A was then directly washed with 60% ethanol. Columns B and C were each eluted with 12 L of 25% ethanol. Finally, each column was eluted with 60% ethanol. The eluates were collected and concentrated separately to obtain a concentrate of the 60% ethanol eluate from columns A, B, and C. Each concentrate was then diluted with purified water to a Brix concentration of approximately 5.0% before undergoing ion exchange treatment.

[0233] The ion exchange resin column is composed of a strong acid cation exchange resin column and a weak base anion exchange resin column connected in series, and the resin loading capacity of each column is 1.8 L. The diluted liquid of the concentrate is sequentially fed into the cation and anion exchange resin columns, wherein the strong acid cation exchange resin model is DOWEX Marathon MSC and the weak base anion exchange resin model is DOWEX Marathon WBA. After the material is fed into the column, the column is washed with purified water. The column flow-through treated with the ion exchange resin and the washing water are combined to obtain a mixed deionized solution. The deionized liquid of each column is concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content are tested. The results are shown in Table 20.

[0234] Table 20

[0235] Comparative Example 2

[0236] 300 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain 1200 L of ultrafiltrate clear liquid a, which was then subjected to multi-stage adsorption separation treatment.

[0237] The multi-stage adsorption separation column set consists of four resin columns, each loaded with 5 L of macroporous adsorption resin (SP700). The columns are sequentially numbered A, B, C, and D, and all columns are regenerated and washed with water for later use. The ultrafiltrate supernatant a is passed through the A, B, C, and D macroporous adsorption resin columns at a flow rate of 3900 ml / h. After the feeding is completed, each column is washed with 10 L of purified water. Column A is then directly washed with 60% ethanol. Columns B, C, and D are each eluted with 10 L of 25% ethanol. Finally, each column is eluted with 60% ethanol. The eluates are collected and concentrated separately to obtain a concentrate of the 60% ethanol eluate from columns A, B, C, and D. Each concentrate is then diluted with purified water to a Brix concentration of approximately 5.0% before undergoing ion exchange treatment.

[0238] The ion exchange resin column consists of a strong acid cation exchange resin column and a weak base anion exchange resin column connected in series, with each column loaded with 1.5 L of resin. The diluted concentrate was sequentially fed into the cation and anion exchange resin columns, where the strong acid cation exchange resin model was DOWEX Marathon MSC and the weak base anion exchange resin model was DOWEX Marathon WBA. After the material was fed into the columns, the columns were washed with purified water. The column flow-through treated with the ion exchange resin and the washing water were combined to obtain a mixed deionized solution. The deionized liquid from each column was concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were tested. The results are shown in Table 21.

[0239] Table 21

[0240] Comparative Example 3

[0241] 280 kg of fresh Momordica grosvenori was crushed into small particles by a crusher and placed in a countercurrent extraction device. 1000 L of hot water at 90° C. was added for extraction. The extract was processed by a juicer, centrifuged, and then clarified by ultrafiltration using a ceramic membrane (pore size 200 nm) to obtain an ultrafiltrate clear liquid a, which was then subjected to multi-stage adsorption separation treatment.

[0242] The multi-stage adsorption separation column set consisted of three resin columns, each loaded with 8 L of macroporous adsorption resin (model XAD16N). The columns were sequentially numbered A, B, and C, and all were regenerated and washed with water before use. The ultrafiltrate supernatant a was passed through the A, B, and C macroporous adsorption resin columns at a flow rate of 3200 ml / h. After the feeding was completed, each column was washed with 16 L of purified water. Column A was then directly washed with 60% ethanol. Columns B and C were each eluted with 16 L of 25% ethanol. Finally, each column was eluted with 60% ethanol. The eluates were collected and concentrated separately to obtain a 60% ethanol eluate concentrate from columns A, B, C, and D. Each concentrate was then diluted with purified water to a Brix concentration of approximately 5.0% before undergoing ion exchange treatment.

[0243] The ion exchange resin column consisted of a strong-acid cation exchange resin column and a weak-base anion exchange resin column connected in series, with each column loaded with 1.8 L of resin. The strong-acid cation exchange resin model was AMBERLITE FPC3500, and the weak-base anion exchange resin model was SCAV2. The ultrafiltrate supernatant a was sequentially passed through the cation and anion exchange resin columns. After the materials were passed through, the columns were washed with purified water. The column flowthrough treated with the ion exchange resin and the wash water were combined to obtain a mixed deionized solution. The deionized liquid from each column was concentrated and dried, and the content of mogroside V and other glycosides, sugar content, and protein content were tested. The results are shown in Table 22.

[0244] Table 22

[0245] The experimental parameter conditions and effect data of each embodiment and comparative example are listed in the following table 23:

[0246] Table 23

[0247] By comparing the above embodiments and comparative examples, it can be seen that the Momordica grosvenori extract was directly subjected to multi-stage adsorption separation without ion exchange treatment and no sweet glycoside composition with a mogroside V content of more than 70% was obtained, indicating that the components of the material that has not been treated with ion exchange are too complex, affecting the separation efficiency of the adsorption resin, and illustrating the necessity of placing the ion exchange treatment step before adsorption separation.

[0248] The effect of extracting mogroside V in the present application is comprehensively evaluated from the two perspectives of the purity of the product obtained and the proportion of high-purity products obtained. The higher the purity of the product obtained, the greater the proportion of high-purity products, and the better the effect. When comparing the separation and purification effects of the embodiments and comparative examples, when the content of mogroside V separated in the last resin column is close, the greater the sum of the net weight proportions of glycoside V obtained in each resin column with a content of mogroside V of more than 70% is, the better the separation and purification effect is; when the sum of the net weight proportions of glycoside V obtained in each resin column with a content of mogroside V of more than 70% is close, the greater the content of mogroside V separated in the last resin column is, the better the separation and purification effect is.

[0249] Experimental example

[0250] 1. Detection methods for various ingredients in monk fruit products

[0251] 1.1 Detection method of mogroside V content

[0252] Mogroside V was tested using the method corresponding to the national standard GB 1886.77-2016;

[0253] 1.2 Detection methods for the content of other glycosides in Momordica grosvenori

[0254] The detection of mogroside components was performed by high performance liquid chromatography external standard method, wherein the chromatographic conditions and sample operation method are as follows:

[0255] 1.2.1 Chromatographic conditions

[0256] 1.2.1.1 Chromatographic column: ZORBAX SB-C18, 4.6×250 mm, 5 μm.

[0257] 1.2.1.2 The mobile phase and its corresponding gradient are shown in Table 24:

[0258] Table 24

[0259] 1.2.1.3 Detector: Diode array UV detector

[0260] 1.2.1.4 Detection wavelength: 203 nm

[0261] 1.2.1.5 Column temperature: 30°C

[0262] 1.2.1.6 Injection volume: 20 μL

[0263] 1.2.2 Sample operation method

[0264] 1.2.2.1 Accurately weigh simenoside I, mogroside IIe, mogroside III, mogroside IVe, 11-O-mogroside V, and mogroside V reference substances, and dissolve them in purified water to prepare four standard solutions containing simenoside I, mogroside IIe, mogroside III, and mogroside IVe at concentrations of 0.015 mg / ml, 0.03 mg / ml, 0.045 mg / ml, and 0.075 mg / ml, respectively; 11-O-mogroside V, 0.015 mg / ml, 0.03 mg / ml, 0.045 mg / ml, and 0.075 mg / ml, respectively. The concentrations of mogroside V were 0.03 mg / ml, 0.06 mg / ml, 0.09 mg / ml and 0.15 mg / ml respectively; the concentrations of mogroside V were 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml and 0.5 mg / ml respectively. The samples were injected and tested according to the above chromatographic conditions. The HPLC spectra of various glycoside components in Momordica grosvenori were shown in FIG1 . The peak area data were collected to prepare the external standard curves of each reference substance.

[0265] 1.2.2.2 Take an appropriate amount of sample, accurately weigh it, dissolve it in ultrapure water and make up to a sample solution of approximately 3 mg / ml. Perform liquid chromatography under the above chromatographic conditions. Collect the chromatogram and identify the components in the sample using the retention time corresponding to each standard. Perform quantitative analysis according to the corresponding standard curve and calculate the content of each component.

[0266] 1.3 Protein content detection method in monk fruit products

[0267] The protein content detection method in the monk fruit product of this application adopts the first method of GB 5009.5-2016.

[0268] 1.4 Detection Method of Total Sugar Content in Monk Fruit Products

[0269] The method for detecting the total sugar content in the monk fruit product of this application adopts the first method of GB 5009.8-2016, HPLC-RID detection.

[0270] 2. Taste test of samples containing mogroside II and mogroside III

[0271] Mogroside II and Mogroside III are representative components of immature Momordica grosvenori and are bitter in taste, with Mogroside II being extremely bitter (Chinese Academy of Sciences, Guangxi Institute of Botany, Research on the Cultivation and Chemistry of Momordica grosvenori, 1st edition, Nanning: Guangxi Science and Technology Press, 2010.5). If the bitterness in the product cannot be controlled at a certain level, it will directly affect the overall taste of the product.

[0272] The samples obtained by elution after separation with resin column A in Example 1 were taken, and after specific purification, they were dissolved in purified water in increasing concentrations. The taste performance was tasted and recorded. The results are shown in Table 25 below.

[0273] Table 25

[0274] The test results in Table 25 above confirm the literature's statement that mogroside II and mogroside III are bitter ingredients, and it is found that even products containing mogroside II and mogroside III still have a significant bitter taste at lower formulation concentrations, indicating that controlling the content of these two ingredients in the product is necessary for product use.

[0275] 3. Comparison of sweetness characteristics of mogroside V and mogroside V

[0276] Samples were taken and separated to obtain a sample of Momordica grosvenori 11-O-glycoside V with a purity of about 77.3%. An appropriate amount was taken and blended to achieve a relative sweetness of 5% relative to sucrose. Another conventional sample that had not been separated and purified (after ion exchange and without multi-column separation and purification) was taken and blended in an appropriate amount to achieve a relative sweetness of 5% relative to sucrose. A taste test was conducted to compare the sweetness peak time and sweetness duration. The results are shown in Table 26.

[0277] Table 26

[0278] In mogroside products, 11-O-mogroside V is beneficial for improving the speed of sweetness response and shortening the aftersweetness to a certain extent, making the overall taste of the sweetener closer to sucrose and improving the eating experience of the product. This shows that obtaining high-purity mogroside V is not necessarily better if the purity is higher. While removing components that have a negative effect on the taste of the product, such as mogroside II and mogroside III, retaining components that promote the overall taste improvement, such as 11-O-mogroside V, and ensuring that the content level is certain is a better combination and process solution.

[0279] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application in any other manner. Any person skilled in the art may utilize the above disclosed technical content to modify or modify the present application into equivalent embodiments with equivalent variations. However, any simple modifications, equivalent variations, and modifications to the above embodiments that do not depart from the technical content of the present application and are based on the technical essence of the present application shall still fall within the scope of protection of the present application.

Claims

1. A sweetening composition, characterized in that: Based on the total weight of the sweetening composition, it comprises: 70% to 85% mogroside V; 5% to 12% 11-O-mogroside V; 0-0.5% simanoside I; 0-0.05% mogroside II and mogroside III; 0-0.5% protein and 0-0.5% total sugars.

2. A method for preparing a sweet composition, characterized in that: The following steps are involved: Extraction: extracting Momordica grosvenori to obtain an extract; Clarifying: clarifying the extract to obtain a clarified solution; Ion exchange treatment: the clarified liquid is sequentially treated with a strong acid cation exchange resin column and a weak base anion exchange resin column to obtain an ion exchange treated liquid; Multi-stage adsorption separation: the ion exchange treated liquid enters the adsorption resin column combination for multi-stage adsorption separation, elution, collection of the eluate, and drying to obtain a sweet composition; Among them, the ion exchange treatment step needs to be carried out before the multi-stage adsorption separation step.

3. The preparation method according to claim 2, characterized in that: The ion exchange treated liquid sequentially enters the first, second, ..., nth (n≥2) adsorption resin columns of the adsorption resin column combination for multi-stage adsorption separation, wherein the separated column liquid obtained after passing through each adsorption resin column directly enters the subsequent adsorption column; or The ion exchange treatment liquid is sequentially passed through the 1st, 2nd, ..., nth (n≥2) adsorption resin columns of the adsorption resin column combination for multi-stage adsorption separation, wherein, after passing through each of the 1st to (x-1) (n≥x≥2) adsorption resin columns, the separated column liquid obtained after passing through each adsorption resin column is collected and combined, and then fed to the next adsorption resin column for separation; then, the column liquid collected after adsorption by the (x-1)th adsorption resin column is sequentially fed into the xth, x+1th, x+2th, ..., nth adsorption resin columns for adsorption separation.

4. The preparation method according to claim 3, characterized in that: After multi-stage adsorption separation, the sweet composition of the present application is obtained from each of the x-th to n-th (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 liquid passing through the 1st, ..., x-1th (2≤x≤n) adsorption resin columns should be 1 to 4 times the sum of the adsorption masses of mogroside V when saturated with the 1st to x-1th adsorption resin columns, preferably 1 to 3 times, and more preferably 1.5 to 3 times.

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, the method further comprises a step of enriching mogroside by using an adsorption resin column A".

7. The preparation method according to claim 6, characterized in that: The adsorption resin column A" is a weak polar macroporous adsorption resin column; Preferably, the adsorption resin column A" is a macroporous adsorption resin column whose framework 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 selected from one of LX-15, D101, AB-8, SP700, XD16N and XD1600N, preferably SP700.

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

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

10. The preparation method according to any one of claims 2 to 9, characterized in that: The particle size of the resin of the strong acid cation exchange resin column is 300 to 1100 μm, preferably 300 to 600 μm; Preferably, the resin of the strong acid cation exchange resin column is selected from one of Marathon MSC, AMBERLITE FPC22H, DIAION SK series and DIAION PK series, 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 selected from one of Marathon WBA, SCAV2, FPA53 and LX-T5, preferably Marathon WBA or SCAV2.

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

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

13. The preparation method according to claims 2 to 12, characterized in that: Based on the total weight of the sweetening composition, the sweetening composition comprises: 70% to 85% mogroside V; 5% to 12% 11-O-mogroside V; 0-0.5% simanoside I; 0-0.05% mogroside II and mogroside III; 0-0.5% protein and 0-0.5% carbohydrates.

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

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