CN117256832A - Preparation method and application of mogroside - Google Patents

Abstract

The invention discloses a preparation method and application of mogroside, wherein the preparation method comprises the following steps: s1.30-90 ℃ fully heating the water extract of the momordica grosvenori to obtain a crude extract; s2, decoloring the crude extract obtained in the step S1; s3, purifying the decolorized solution obtained in the step S2 by using macroporous adsorption resin, and collecting the decolorized solution in sections by using ethanol as eluent; s4, concentrating. The invention can obtain most of mogrosides in the raw materials only by one time of extraction, does not need to use other organic solvents to improve the extraction rate, has simple and convenient operation and low cost, and can be used for industrial production and processing of foods. The momordica grosvenori sweet glycoside and the enzymolysis momordica grosvenori sweet glycoside provided by the invention have a taste similar to that of sucrose, can be used as a sweetener for replacing sucrose, are suitable for different types of foods, and can meet different production demands of markets and enterprises.

Description

A kind of preparation method and application of mogroside

Technical field

The present invention relates to raw materials and additives in the food field. Specifically, it relates to a preparation method of mogroside and its application.

Background technique

Sweetness is one of the basic human tastes. Sweetness is the taste of happiness that countless people cannot resist. Substances in food that can provide sweetness include nutritive sweeteners and non-nutritive sweeteners. Nutritive sweeteners include white sugar, fructose syrup, crystallized fructose, etc.; non-nutritive sweeteners are divided into synthetic sweeteners and natural sweeteners. Synthetic sweeteners include sucralose, acesulfame potassium, and aspergillus. Batame, etc.; natural sweeteners include steviol glycosides, mogrosides, etc. Bulk sweeteners can provide solid taste and low energy value. They are mostly sugar alcohols, including erythritol, xylitol, maltitol, etc.

Non-nutritive sweeteners generally have low or no caloric value and can effectively reduce cardiovascular and cerebrovascular diseases, dental caries and other diseases caused by excessive sugar intake. In addition, non-nutritive sweeteners can also meet the sweetness requirements of some patients who are not suitable for consuming sucrose, especially those with abnormal sugar metabolism. In order to meet consumers' health needs to reduce sugar intake, non-nutritive sweeteners gradually replace nutritional sweeteners. However, the safety and stability issues of synthetic sweeteners have limited their application to a certain extent. For example, saccharin has certain toxicity and is a potential carcinogen, and has been banned or restricted in many countries. However, traditional natural sweeteners such as steviol glycosides and sweet tea glycosides generally have obvious bitterness and strong plant flavor. They have insufficient front sweetness and too long after-sweetness, so their acceptance by consumers is not high.

The preparation process of natural sweeteners includes plant extraction and biotransformation. The natural sweetener prepared from Luo Han Guo, which is used both as medicine and food, is an ideal sucrose substitute. It not only has high sweetness and low calories, but also has the effects of enhancing immunity, anti-fatigue and lowering blood sugar. It is not absorbed by the body after consumption and can Effectively reduce the intake of energy substances. However, in the prior art, Luo Han Guo is used to prepare natural sweeteners, but there are problems such as product odor, product loss, product instability, high cost, and low economic value.

Contents of the invention

In order to solve the problems of complex preparation process and lack of taste of natural sweeteners in the prior art, the present invention provides a preparation method of mogroside and its application. By optimizing the preparation process of natural sweeteners and different sweetness The compounding technology of raw materials improves the overall taste of the sweetener, making it closer to sucrose in taste than similar products.

The first object of the present invention is to provide a method for preparing mogroside.

The second object of the present invention is to provide mogroside prepared by the above method.

The third object of the present invention is to provide the application of the above-mentioned mogroside as a sweetener in food preparation and/or processing.

The fourth object of the present invention is to provide a method for enzymatic hydrolysis of mogroside.

The fifth object of the present invention is to provide products prepared by the above method.

The sixth object of the present invention is to provide the application of the above product as a sweetener in food preparation and/or processing.

The seventh object of the present invention is to provide a sweetener.

In order to achieve the above objects, the present invention is achieved through the following solutions:

By adjusting the extraction process materials and parameters, the present invention obtains mogroside with pure sweetness through a segmented extraction method. Afterwards, compounding technology was used to optimize the odor (bitterness, licorice taste, etc.) and sweetness of the raw materials.

The present invention searches for the mogroside extract with the best sweetness through sensory evaluation during the decolorization, deodorization and enrichment stages. The finally selected extract has a pure sweetness and no foreign taste, is convenient and fast, and can be directly used for the development of subsequent products. The extracts in the remaining stages can be recycled to enrich high-quality extracts, or reprocessed for other applications.

A method for preparing mogroside, comprising the following steps:

S1. Fully heat the aqueous extract of Luo Han Guo at 30°C to 90°C to obtain a crude extract;

S2. Use macroporous ion exchange resin to decolorize the crude extract obtained in step S1, elute with water, measure the sweetness of the eluate, and use the sweetness lower than 1% (w/v) sucrose aqueous solution as the elution end point. Collect the resulting eluate to obtain decolorizing liquid;

S3. Use macroporous adsorption resin to purify the decolorized liquid obtained in step S2, use 50% (v/v) to 90% (v/v) ethanol to elute, and collect the eluate in sections to obtain the purified liquid; The method of segmented collection is as follows: according to 0.2 column volume to 0.5 column volume being 1 part, collect the 5th to 8th part of the eluent from the beginning of elution;

S4. Concentrate the purified liquid obtained in step S3 to obtain mogroside.

The present invention has no special restrictions on the variety, source and processing technology of Luo Han Guo raw materials. The processing technology of Luo Han Guo includes but is not limited to freeze-drying, air-drying and baking.

The peel of freeze-dried monk fruit is easy to remove. Compared with fresh monk fruit and baked monk fruit, it not only solves the problem of ordinary fresh monk fruit being unable to be stored for a long time and the raw material planting cycle, but also solves the problem of baked monk fruit having strong odor and burdening the decolorization process. Big question. The extracted raw materials retain the kernels with the highest mogroside content and relatively few impurities.

Preferably, the Luo Han Guo is freeze-dried Luo Han Guo.

Preferably, in step S1, the aqueous extract of Luo Han Guo is obtained by ultrasonic treatment at 20 KHz to 100 KHz for 10 to 20 minutes with an aqueous solution of Luo Han Guo powder.

More preferably, in step S1, the ultrasonic method is: ultrasonic treatment at 40 KHz for 10 minutes.

More preferably, in step S1, the preparation method of the monk fruit powder is: after removing the shell of the monk fruit, crush it and sieve.

Further preferably, in step S1, the preparation method of the Luo Han Guo powder is: after removing the shell from the Luo Han Guo, crush it through a wall breaker and pass through a 200 mesh sieve.

More preferably, in step S1, in the aqueous solution containing monk fruit powder, the material-to-liquid ratio of monk fruit powder and water is 1: (10-20).

More preferably, in step S1, in the aqueous solution containing monk fruit powder, the material-to-liquid ratio of monk fruit powder and water is 1:15.

Preferably, in step S1, heating is performed at 30-90°C for 60-120 minutes.

More preferably, in step S1, heat at 30°C for 90 minutes.

Preferably, in step S1, the heating method is water bath heating.

More preferably, in step S1, stirring is also required during the heating process, and the stirring rate is 200 rpm to 300 rpm.

Further preferably, in step S1, stirring is also required during the heating process, and the stirring rate is 200 rpm.

Preferably, in step S1, centrifugation is required after heating, and the centrifugation method is: centrifugation at 6,000 rpm to 10,000 rpm for 5 min to 10 min.

More preferably, in step S1, the centrifugation method is: centrifuge at 10,000 rpm for 10 minutes.

Preferably, in step S2, the macroporous ion exchange resin is any one of D280 macroporous anion exchange resin, LSA-700 macroporous anion exchange resin or D941 macroporous anion exchange resin.

More preferably, in step S2, the macroporous ion exchange resin is D280 resin.

Preferably, in step S2, the decolorization includes feeding and elution.

More preferably, in step S2, the feed flow rate for decolorization is 2 mL/min to 4 mL/min.

Further preferably, in step S2, the feed flow rate for decolorization is 3 mL/min.

More preferably, in step S2, the elution is to flush the macroporous ion exchange resin with 2 column volumes of purified water at a flow rate of 2 mL/min to 4 mL/min, and is prepared according to the method for measuring sweetness in GB 8270-1999 1% (w/v) sucrose aqueous solution. Sensory evaluation personnel were trained and screened in accordance with GB/T 16291.2 2010. 1% (w/v) sucrose aqueous solution was used as the standard solution. The resulting eluate was sensory evaluated by the sensory evaluation personnel until it was discharged. When the sweetness of solution 1 is lower than the standard solution, it is the elution end point. The resulting eluate is collected to obtain the decolorized solution.

Further preferably, in step S2, the elution is to rinse the macroporous ion exchange resin with 2 column volumes of purified water at a flow rate of 3mL/min, and prepare 1% (w) according to the method for measuring sweetness in GB 8270-1999 /v) sucrose aqueous solution, the sensory evaluation personnel were trained and screened in accordance with GB/T 16291.2 2010, and 1% (w/v) sucrose aqueous solution was used as the standard solution. The resulting eluate was sensory evaluated by the sensory evaluation personnel until the sweetness of the effluent 1 The elution end point is when the concentration is lower than the standard solution, and the resulting eluate is collected to obtain the decolorizing solution.

Preferably, in step S3, the macroporous adsorption resin is any one of D101 resin, AB-8 resin or D941 resin.

More preferably, in step S3, the macroporous adsorption resin is D101 resin.

Preferably, in step S3, before elution, the macroporous adsorption resin is washed with 2 column volumes of purified water at a flow rate of 2 mL/min to 4 mL/min.

Preferably, in step S3, 75% (v/v) ethanol is used as the eluent.

Preferably, in step S3, the method of collecting the eluate in stages is: taking 0.32 column volumes as 1 part, collecting the 6th part of the eluate since the start of elution.

Preferably, in step S3, the concentration method is vacuum concentration under reduced pressure.

Preferably, in step S3, the concentration method is rotary evaporation under vacuum at 60°C in a water bath and 0.098Mpa until evaporated to dryness.

Mogroside prepared by any of the above methods should also be within the protection scope of the present invention.

The application of mogroside prepared by any of the above methods as a sweetener in food preparation and/or processing should also be within the protection scope of the present invention.

Preferably, the food is a drink.

More preferably, the beverage is tea.

The present invention uses enzymatic hydrolysis technology to modify the odor of mogroside and uses a compounding process to mix different ingredients to create a synergistic effect with each other, thereby making the taste closer to sucrose in the complete process of showing sweetness, especially the unique characteristics of mogroside. The cooling feeling, astringency and trailing feeling are significantly improved.

A method for enzymatically hydrolyzing mogroside. Maltodextrin and the mogroside are fully mixed evenly, and the resulting mixture is fully enzymatically hydrolyzed by glycosyltransferase.

Preferably, the mass ratio of mogroside and maltodextrin is 1: (1-3).

More preferably, the mass ratio of mogroside and maltodextrin is 1:2.

Preferably, the mass-to-volume ratio of mogroside and glycosyltransferase is 1 g: (0.01-0.02) mL.

More preferably, the method for sufficient enzymatic hydrolysis is: fully mix mogroside and maltodextrin according to the mass ratio of 1: (1 to 3), and the resulting mixture is based on the mass to volume ratio of mogroside and glycosyltransferase. 1g: (0.01～0.02)mL, mix thoroughly and react at 50℃～60℃ for 100min～150min.

Further preferably, the sufficient reaction method is: fully mix the above-mentioned mogroside and maltodextrin according to a mass ratio of 1:2, and the resulting mixture has a mass volume of 1g:0.0125mL of mogroside and glycosyltransferase. Mix thoroughly and react at 60°C for 120 minutes.

More preferably, the glycosyltransferase is ɑ-Amylase G "Amano" L liquid enzyme of Amano Enzyme Inc. Shanghai Branch, EINECS NO. 232-565-6.

Further preferably, the mass-volume ratio of mogroside and ɑ-Amylase G "Amano" L liquid enzyme is 1g:0.0125mL.

More preferably, the method for sufficient enzymatic hydrolysis is: fully mix the mogroside and maltodextrin according to a mass ratio of 1: (1-3), and the resulting mixture is based on the ratio of mogroside and ɑ-Amylase G" Amano"L liquid enzyme is mixed thoroughly according to the mass-volume ratio of 1g:(0.01~0.02)mL, and reacted at 50℃~60℃ for 100min~150min.

Most preferably, the sufficient reaction method is: fully mix the mogroside and maltodextrin in a mass ratio of 1:2, and the resulting mixture is in accordance with the formula of mogroside and ɑ-Amylase G "Amano" L liquid enzyme The mass-to-volume ratio is 1g:0.0125mL, mix thoroughly, and react at 60°C for 120 minutes.

Preferably, the method further comprises heat inactivating the glycosyltransferase.

More preferably, the heating inactivation method is heating at 80°C to 95°C for 10 to 20 minutes.

Further preferably, the heating inactivation method is heating at 90°C for 10 minutes.

Products prepared by any of the above methods should also be within the protection scope of the present invention.

The application of the product prepared by any of the above methods as a sweetener in food preparation and/or processing should also be within the protection scope of the present invention.

Preferably, the food is a drink.

More preferably, the drink is coconut milk.

Further preferably, the coconut milk is sugar-free coconut milk.

A sweetener comprising any of the above-mentioned mogrosides and/or any product prepared by any of the above-mentioned methods.

Preferably, the sweetener further contains any one of thaumatin, erythritol or arabinose.

More preferably, the sweetener consists of a product prepared by any of the above methods and arabinose.

Further preferably, the sweetener is composed of 1 to 10 parts by weight of the product prepared by any of the above methods and 1 to 500 parts by weight of arabinose.

More preferably, the sweetener is composed of 1 to 10 parts by weight of the product prepared by any of the above methods and 300 to 400 parts by weight of arabinose.

More preferably, the sweetener is composed of 1 part by weight of the above-mentioned mogroside and 300-400 parts by weight of arabinose.

Most preferably, the sweetener consists of 1 part by weight of the product produced by any method and 333.3 parts by weight of arabinose.

The application of the above-mentioned sweeteners in food manufacturing and/or food processing should also be within the protection scope of the present invention.

Preferably, the food is a drink.

More preferably, the drink is coconut milk.

Further preferably, the coconut milk is sugar-free coconut milk.

Compared with the prior art, the present invention has the following beneficial effects:

The method only needs one extraction to obtain most of the mogrosides in the raw materials, and does not need to use other organic solvents to increase the extraction rate. It is easy to operate, has low cost, and can be used for industrial production and processing of food.

The present invention contains mogroside and enzymatically hydrolyzed mogroside, has a taste that is highly similar to that of sucrose, can be used as a sweetener to replace sucrose, is suitable for different types of food, and can meet the different production needs of the market and enterprises. .

Description of the drawings

Figure 1 shows the sensory evaluation results of mogroside used as a sweetener.

Figure 2 shows the sensory evaluation results of a composition containing mogroside used as a sweetener.

Detailed ways

The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments of the description. The embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention. The test methods used in the following examples are conventional methods unless otherwise stated; the materials and reagents used, unless otherwise stated, are commercially available reagents and materials. The volume of one column in the examples of the present invention is equivalent to 125 mL.

Embodiment 1 A method for preparing mogroside

1. Raw material pretreatment

Select dried monk fruit raw materials, remove the shell, take 10g and crush it with a wall breaker, and pass it through a 200 mesh sieve to obtain powdered raw materials.

2. Extraction

Mix the powdery raw material obtained in step 1 with pure water, the material-to-liquid ratio is 1:15; perform ultrasonic extraction pretreatment for 10 minutes, and the ultrasonic frequency is 40KHz.

Then put it into a water bath that has been preheated to 30°C for stirring and extraction for 90 minutes. The stirring speed is 200 rpm.

Use a centrifuge to centrifuge at 10,000 rpm for 10 minutes and collect the supernatant to obtain a crude extract.

3. Decolorization

Use a chromatography column containing D280 macroporous anion exchange resin for decolorization, and control the feed flow rate to 3 mL/min; after the feed is completed, use 2 column volumes of pure water to flush the D280 resin at 3 mL/min.

Prepare 1% (w/v) sucrose aqueous solution according to the method for measuring sweetness in GB 8270-1999; train and screen sensory evaluation personnel according to GB/T16291.2 2010, use 1% (w/v) sucrose aqueous solution as the standard solution, The obtained eluate was subjected to sensory evaluation by an evaluator until the sweetness of the aqueous eluate was lower than 1% (w/v) sucrose aqueous solution, which was the elution end point.

Collect the water wash to obtain the decolorized extract.

4. Purification

Purify the decolorized extract obtained in step 3 through D101 macroporous adsorption resin, and control the feed flow rate to 3mL/min; after the feed is completed, use 2 column volumes of pure water to rinse the D101 resin at 3mL/min, and use 75% Elute with (v/v) ethanol. Starting from adding 75% (v/v) ethanol, collect the eluate in stages according to each 0.32 column volume as a stage. A total of 8 stages of eluate are collected.

5. Concentrate

The 8 portions of the eluate collected in step 4 were concentrated and rotary evaporated under vacuum at 60°C and 0.098Mpa in a water bath until evaporated to dryness, and 8 portions of the concentrate were collected.

6. Sweetness measurement

Sensory evaluation personnel are trained and screened in accordance with GB/T 16291.2 2010, and tasted sweetener monomer samples including but not limited to commercially available steviol glycosides, erythritol, mogrosides, and arabinose when they are sweeter than 10% (w/v ) sensory characteristics of sucrose solution, and screened out the more common characteristic lexicon for mainstream sweeteners, such as sweetness, bitterness, astringency, coolness, grassy taste, metallic feeling and tailing feeling (sweetness). The lingering feeling of taste in the mouth) and other sensory descriptions.

Dissolve 8 portions of the concentrate obtained in the previous step with 40 mL of pure water. Prepare a 10% (w/v) sucrose aqueous solution according to the method for measuring sweetness in GB 8270-1999; use 10% (w/v) sucrose aqueous solution as the standard solution. Evaluate the sweetness and off-flavor of 8 concentrated liquids collected in sections. The specific judgment criteria are as follows:

Sweetness judgment: After evaluation by the sensory evaluator, the sweetness of the liquid to be tested is significantly lower than 10% (w/v) sucrose water and close to RO water, then the sweetness is judged to be no sweetness; after evaluation by the sensory evaluator, the sweetness of the liquid to be tested is If the sweetness of the liquid to be tested is close to or equal to 10% (w/v) sucrose water, the sweetness is judged to be slightly sweet; after evaluation by the sensory evaluator, if the sweetness of the liquid to be tested is significantly higher than 10% (w/v) sucrose water, the sweetness is judged to be slightly sweet. The sweetness is very sweet;

Judgment of peculiar smell: After evaluation by the sensory evaluator, if the liquid to be tested is close to RO water and has no obvious peculiar smell, then the peculiar smell is judged to be no peculiar smell; after evaluation by the sensory evaluator, if the liquid to be tested has a slight bitter, cool or grassy smell, then the smell is judged to be zero. The odor is judged to be light; if the test liquid has obvious bitter, cool or grassy odor after evaluation by the sensory evaluation personnel, the odor is judged to be heavy.

Table 1 shows the measurement results of the sweetness and off-flavor of 8 concentrated solutions.

Table 1 Measurement results of sweetness and off-flavor of the concentrated liquid of Example 1

The results show that the sweetest among the 8 concentrates is the 6th concentrate, that is, the eluent from the 6th stage (201mL~240mL) collected in stages, as the mogroside finally obtained in this example .

Example 2 A method for preparing mogroside

1. Raw material pretreatment

Same as step 1 of Example 1.

2. Extraction

Mix the powdery raw material obtained in step 1 with pure water, the material-to-liquid ratio is 1:15; perform ultrasonic extraction pretreatment for 15 minutes, and the ultrasonic frequency is 40KHz.

Then put it into a water bath that has been preheated to 90°C for stirring and extraction for 120 minutes, and the stirring speed is 200 rpm.

Use a centrifuge to centrifuge at 10,000 rpm for 10 minutes and collect the supernatant to obtain a crude extract.

3. Decolorization

Use a chromatography column containing D280 macroporous anion exchange resin for decolorization, and control the feed flow rate to 3 mL/min; after the feed is completed, use 2 column volumes of purified water to flush the D280 resin at 3 mL/min.

Evaluate the sweetness of the washing liquid according to step 3 of Example 1 until the washing liquid has no sweetness as the rinse end point.

Collect the water wash to obtain the decolorized extract.

4. Purification

Purify the decolorized extract obtained in step 3 through D101 macroporous adsorption resin, and control the feed flow rate to 3mL/min; after the feed is completed, use 2 column volumes of purified water to rinse the D101 resin at 4mL/min, and use 75% (v/v) ethanol is used to elute D101 resin. Starting from adding 75% (v/v) ethanol, the eluate is collected in stages according to every 0.4 column volumes. A total of 6 stages of elution are collected. liquid.

5. Concentrate

Concentrate 6 parts of the eluent according to step 5 of Example 1 to obtain 6 parts of concentrate.

6. Sweetness measurement

According to step 6 of Example 1, the sweetness and off-flavor of 6 concentrated solutions were measured, and the results are shown in Table 2.

Table 2 Measurement results of sweetness and off-flavor of the concentrate of Example 2

The results show that the sweetest among the 6 concentrates is the 5th concentrate, that is, the eluate from the 5th stage (201mL~250mL) collected in stages, as the mogroside finally obtained in this example .

Embodiment 3 A method for preparing mogroside

1. Raw material pretreatment

Same as step 1 of Example 1.

2. Extraction

Mix the powdery raw material obtained in step 1 with pure water, the material-liquid ratio is 1:20; perform ultrasonic extraction pretreatment for 20 minutes, and the ultrasonic frequency is 80KHz.

Then put it into a water bath that has been preheated to 30°C for stirring and extraction for 60 minutes, and the stirring speed is 200 rpm.

Use a centrifuge to centrifuge at 10,000 rpm for 10 minutes and collect the supernatant to obtain a crude extract.

3. Decolorization

Use a chromatography column containing D280 macroporous anion exchange resin for decolorization, and control the feed flow rate to 4 mL/min; after the feed is completed, use 2 column volumes of purified water to flush the D280 resin at 4 mL/min.

Evaluate the sweetness of the washing liquid according to step 3 of Example 1 until the washing liquid has no sweetness as the rinse end point.

Collect the water wash to obtain the decolorized extract.

4. Purification

Purify the decolorized extract obtained in step 3 through AB-8 macroporous adsorption resin, and control the feed flow rate to 4 mL/min; after the feed is completed, use 2 column volumes of purified water to flush the AB-8 resin at 2 mL/min. , use 75% (v/v) ethanol to elute the AB-8 resin. Starting from adding 75% (v/v) ethanol, collect the eluent in stages according to every 0.24 column volumes. A total of 10 stages of eluent.

5. Concentrate

Concentrate 10 parts of the eluate according to step 5 of Example 1 to obtain 10 parts of concentrate.

6. Sweetness measurement

According to step 6 of Example 1, the sweetness and off-flavor of 10 portions of the concentrated liquid were measured, and the results are shown in Table 3.

Table 3 Measurement results of sweetness and off-flavor of the concentrate of Example 3

The results show that the sweetest one among the 10 concentrates is the 8th concentrate, which is the eluate from the 8th stage (211mL~240mL) collected in stages, as the mogroside finally obtained in this example .

Embodiment 4 A method for preparing mogroside

1. Raw material pretreatment

Same as step 1 of Example 1.

2. Extraction

Mix the powdery raw material obtained in step 1 with pure water, the material-to-liquid ratio is 1:10; perform ultrasonic extraction pretreatment for 20 minutes, and the ultrasonic frequency is 40KHz.

Then put it into a water bath that has been preheated to 60°C for stirring and extraction for 120 minutes, and the stirring speed is 200 rpm.

Use a centrifuge to centrifuge at 10,000 rpm for 10 minutes and collect the supernatant to obtain a crude extract.

3. Decolorization

Use a chromatography column containing LSA-700 macroporous anion exchange resin for decolorization, and control the feed flow rate to 3 mL/min; after the feed is completed, use 2 column volumes of purified water to flush the LSA-700 resin at 3 mL/min.

Evaluate the sweetness of the washing liquid according to step 3 of Example 1 until the washing liquid has no sweetness as the rinse end point.

Collect the water wash to obtain the decolorized extract.

4. Purification

Purify the decolorized extract obtained in step 3 through AB-8 macroporous adsorption resin, and control the feed flow rate to 3 mL/min; after the feed is completed, use 2 column volumes of purified water to flush the AB-8 resin at 2 mL/min. , use 90% (v/v) ethanol to elute the AB-8 resin. Starting from adding 90% (v/v) ethanol, collect the eluent in stages according to every 0.24 column volumes. A total of 10 stages of eluent.

5. Concentrate

Concentrate 10 parts of the eluate according to step 5 of Example 1 to obtain 10 parts of concentrate.

6. Sweetness measurement

According to step 6 of Example 1, the sweetness and off-flavor of 10 portions of the concentrated solution were measured, and the results are shown in Table 4.

Table 4 Measurement results of sweetness and off-flavor of the concentrated liquid of Example 4

The results show that the sweetest one among the 10 concentrates is the 8th concentrate, which is the eluate from the 8th stage (211mL~240mL) collected in stages, as the mogroside finally obtained in this example .

Embodiment 5 A method for preparing mogroside

1. Raw material pretreatment

Same as step 1 of Example 1.

2. Extraction

Mix the powdery raw material obtained in step 1 with pure water, the material-to-liquid ratio is 1:15; perform ultrasonic extraction pretreatment for 10 minutes, and the ultrasonic frequency is 40KHz.

Then put it into a water bath that has been preheated to 60°C for stirring and extraction for 120 minutes, and the stirring speed is 200 rpm.

Use a centrifuge to centrifuge at 10,000 rpm for 10 minutes and collect the supernatant to obtain a crude extract.

3. Decolorization

Use a chromatography column containing D941 macroporous anion exchange resin for decolorization, and control the feed flow rate to 3 mL/min; after the feed is completed, use 2 column volumes of purified water to flush the D941 resin at 3 mL/min.

Evaluate the sweetness of the washing liquid according to step 3 of Example 1 until the washing liquid has no sweetness as the rinse end point.

Collect the water wash to obtain the decolorized extract.

4. Purification

Purify the decolorized extract obtained in step 3 through ADS-7 macroporous adsorption resin, and control the feed flow rate to 3mL/min; after the feed is completed, use 2 column volumes of purified water to flush the ADS-7 resin at 3mL/min. , use 75% (v/v) ethanol to elute the ADS-7 resin. Starting from adding 75% (v/v) ethanol, collect the eluent in stages according to every 0.32 column volumes. A total of 8 stages of eluent.

5. Concentrate

Concentrate 8 parts of the eluate according to step 5 of Example 1 to obtain 8 parts of concentrate.

6. Sweetness measurement

According to step 6 of Example 1, the sweetness and off-flavor of 8 portions of the concentrated liquid were measured, and the results are shown in Table 5.

Table 5 Measurement results of sweetness and off-flavor of the concentrated liquid of Example 5

The results show that the sweetest among the 8 concentrates is the 6th concentrate, that is, the eluent from the 6th stage (201mL~240mL) collected in stages, as the mogroside finally obtained in this example .

Application Example 1 Mogroside used as sweetener

1. Experimental methods

1. Sweetness calibration of sweeteners

The mogroside prepared in Example 1 and the commercially available monk fruit extract (mogroside V50, CAS No.: 88901-36-4) and steviol glycoside RA97% (CAS No.: 58543-16-1) were used as the sweetener to be tested. Flavor agent, using the sweetness of 10% (w/v) sucrose aqueous solution at 20°C as a control (recorded as "sweetness is 1.0"), perform sensory evaluation and measurement according to step 6 in Example 1. The sweetness to be tested The agent was subjected to sensory evaluation and sweetness evaluation, and the amount of the sweetener to be tested in pure water was calibrated based on the results. The specific situation is shown in Table 6.

Table 6 Sweetness calibration situation of application example 1

2. Preparation of products to be tested

Soak tea leaves and boiling water at a ratio of (w/v) 1:50 for 5 minutes to obtain a tea solution.

According to the sweetness calibration results shown in Table 6, add 0.04% (w/v) mogroside prepared in Example 1 to the tea solution, and mix thoroughly to obtain product 1 to be tested. Add 0.05% to the tea solution. % (w/v) commercially available Luo Han Guo extract (mogroside V50, CAS number: 88901-36-4), mix thoroughly to obtain the product to be tested 2; add 0.05% (w/v) to the tea solution Steviol glycoside RA97% (CAS number: 58543-16-1), mix thoroughly to obtain the product to be tested 3; add 10% (w/v) sucrose (CAS number: 57-50-1) to the tea solution, Mix thoroughly to obtain the control product.

3. Sensory evaluation

According to the regulations of GB/T 16291.1&2 2010, sensory evaluation personnel are trained and selected to conduct sensory evaluation of sweetness and off-flavor of the products to be tested 1 to 5 based on the control product.

The evaluation criteria are as follows:

Sweetness: the lowest score is 0 points, the highest score is 15 points;

Bitterness: the lowest score is 0 points, the highest score is 15 points;

Green grass feeling: the lowest score is 0 points, the highest score is 15 points;

Cooling feeling: the lowest score is 0 points, the highest score is 15 points;

Astringency: the lowest score is 0 points, the highest score is 15 points;

When conducting sensory evaluation of the above products to be tested, a straight line with a length of 15 cm is used as a scale. The left end of the scale indicates "none or the smallest" evaluation characteristics, and the right end of the scale indicates the "strongest or maximum" evaluation characteristics. The tested samples were compared with each other to select the main features and marked their intensity with a scale. The characteristics suitable for tea are sweetness, bitterness, grassy taste, coolness and astringency.

2. Experimental results

Table 7 Sensory evaluation results of application example 1

As shown in Table 7 and Figure 1, using the method of Example 1 of the present invention, the mogroside obtained through segmented collection is used as a sweetener and added to tea. The sweetness is closer to sucrose. Compared with other The product to be tested also had reduced astringency, bitterness and grassiness to varying degrees, and added a unique and pleasant honey-like flavor.

Example 6 An enzymatic hydrolysis method of mogroside

The enzymatic hydrolysis method provided in this example is suitable for mogroside prepared by any of the methods described in Examples 1 to 5 or by conventional methods in this field.

Taking mogroside prepared in Example 1 of the present invention as an example, the specific operating steps are as follows:

1. Raw material pretreatment

The mogroside and maltodextrin (CAS number: 9050-36-6) prepared in Example 1 were thoroughly mixed and dissolved according to a mass ratio of 1:2 to obtain the enzymatic hydrolysis raw material.

2. Enzymatic hydrolysis

According to the mass and volume ratio of mogroside and glycosyltransferase = 1g:0.0125mL, add glycosyltransferase (i.e., ɑ- from Amano Enzyme Inc. Shanghai Branch) to the enzymatic hydrolysis raw material obtained in step 1. Amylase (G "Amano" L liquid enzyme) is enzymatically hydrolyzed in a water bath at a temperature of 60°C for 120 minutes to obtain the enzymatic hydrolysis product.

3. Inactivation

Place the enzymatic hydrolysis product obtained in step 2 in a 90°C water bath for 10 minutes to fully inactivate the glycosyltransferase, thereby obtaining enzymatically hydrolyzed mogroside.

Example 7 An enzymatic hydrolysis method of mogroside

The enzymatic hydrolysis method provided in this example is suitable for mogroside prepared by any of the methods described in Examples 1 to 5 or by conventional methods in this field.

Taking mogroside prepared in Example 1 of the present invention as an example, the specific operating steps are basically the same as those in Example 6, and the only difference is:

In step 1, the mogroside and maltodextrin (CAS number: 9050-36-6) prepared in Example 1 are thoroughly mixed and dissolved according to the mass ratio of 1:3; in step 2, mogroside and glycosyltransferase (i.e. ɑ-Amylase G "Amano" L liquid enzyme from AmanoEnzyme Inc. Shanghai Branch) was mixed at a mass/volume ratio of 1:0.05 for enzymatic hydrolysis; the temperature was 50°C, and the reaction time was 240min; in step 3, Place the enzymatic hydrolysis product obtained in step 2 in a 90°C water bath for 20 minutes.

Application Example 2: A composition containing mogroside is used as a compound sweetener

1. Experimental methods

The enzymatic mogroside provided by the invention can be combined with conventional commercial sweeteners (such as thaumatin (CAS No.: 71396-29-7), erythritol (CAS No.: 149-32-6) and Arabinose (CAS No.: 147-81-9)) is used for compounding. The following takes arabinose (CAS No.: 147-81-9) as an example to provide a composition containing mogroside and its use for compounding. Specific uses of sweeteners.

1. Preparation of compound sweeteners

The enzymatic mogroside prepared in Example 6 of the present invention was used as the main material, and arabinose (CAS No.: 147-81-9) was used as the auxiliary material. The mass ratio of the main material to the auxiliary material was 1:333.3, and the mixture was evenly mixed. That is, compound sweetener 1 (containing the enzymatic mogroside prepared in Example 6) was obtained.

2. Sweetness calibration of compound sweeteners

Using sugar-free coconut milk as the test matrix, compound sweetener 1, mogroside prepared in Example 1, and commercially available monk fruit extract (mogroside V50, CAS number: 88901-36-4) and stevia Glycoside RA97% (CAS number: 58543-16-1) was used as the sweetener to be tested, and a 5% (w/v) sucrose aqueous solution was used as a control with a sweetness of 0.5 as measured by the evaluator at 20°C. According to the application example Step 1 in 1 carries out sweetness calibration, the details are shown in Table 8.

Table 8 Sweetness calibration situation of application example 2

3. Preparation of products to be tested

According to the sweetness calibration results shown in Table 8, add 4.012% (w/v) compound sweetener 1 to the sugar-free coconut milk and mix thoroughly to obtain the product to be tested 1; add 0.02% (w/v) mogroside of Example 1, mix thoroughly to obtain the product to be tested 2; add 0.025% (w/v) commercially available monk fruit extract (mogroside V50, 88901-36-4), mix thoroughly to obtain the product to be tested 3; add 0.025% (w/v) steviol glycoside RA97% (58543-16-1) to the sugar-free coconut milk, mix thoroughly to obtain Product to be tested 4: Add 5% (w/v) sucrose (CAS number: 57-50-1) to sugar-free coconut milk, mix thoroughly to obtain the control product.

4. Sensory evaluation

According to the "sensory evaluation" method in Application Example 1, using the control product of this Application Example as the standard, conduct sensory evaluation of sweetness and off-flavor of the products to be tested 1 to 5 of this Application Example.

2. Experimental results

Table 9 Sensory evaluation results of application example 2

As shown in Table 9 and Figure 2, compared with other products to be tested, the compound composition of mogroside and arabinose (compound sweetener 1) enzymatically hydrolyzed by the method of Example 6 of the present invention has better effects on tailing. The taste, coolness, astringency and bitterness all have a significant modification effect, making up for part of the lack of taste, making it taste closer to sucrose in the complete process of showing sweetness.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and do not limit the protection scope of the present invention. For those of ordinary skill in the art, based on the above descriptions and ideas, they can also make There are other variations or modifications in different forms, and it is not necessary and impossible to exhaustively enumerate all implementations here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention shall be included in the protection scope of the claims of the present invention.

Claims (10)

1. A method for preparing mogroside, characterized in that it includes the following steps: S1. Fully heat the aqueous extract of Luo Han Guo at 30°C to 90°C to obtain a crude extract; S2. Use macroporous ion exchange resin to decolorize the crude extract obtained in step S1, elute with water, measure the sweetness of the eluate, and use the sweetness lower than 1% (w/v) sucrose aqueous solution as the elution end point. Collect the resulting eluate to obtain decolorizing liquid; S3. Use macroporous adsorption resin to purify the decolorized liquid obtained in step S2, use 50% (v/v) to 90% (v/v) ethanol to elute, and collect the eluate in sections to obtain the purified liquid; The method of segmented collection is as follows: according to 0.2 column volume to 0.5 column volume being 1 part, collect the 5th to 8th part of the eluent from the beginning of elution; S4. Concentrate the purified liquid obtained in step S3 to obtain mogroside. 2. The method according to claim 1, characterized in that in step S1, heating is performed at 30°C to 90°C for 60 to 120 minutes. 3. The method according to claim 1, characterized in that, in step S1, the aqueous extract of Luo Han Guo is obtained from the aqueous solution of Luo Han Guo powder by ultrasonic treatment at 20 KHz to 100 KHz for 10 to 20 minutes. 4. Mogroside prepared by the method of any one of claims 1 to 3. 5. The application of mogroside as claimed in claim 4 as a sweetener in food production and/or processing. 6. A method for enzymatic hydrolysis of mogroside, characterized in that maltodextrin and mogroside according to claim 4 are fully mixed evenly, and the resulting mixture is fully enzymatically hydrolyzed by glycosyltransferase. 7. The method according to claim 6, characterized in that the mass ratio of the maltodextrin and the mogroside of claim 4 is 1: (1-3). 8. The product prepared by the method of any one of claims 6 to 7. 9. Application of the product of claim 8 as a sweetener in food preparation and/or processing. 10. A sweetener, characterized by comprising the mogroside of claim 4 and/or the product of claim 8.