Preparation method of sugarcane polyphenol with caramel taste

Abstract

The invention relates to the technical field of natural active substance extraction, in particular to a preparation method of sugarcane polyphenol with caramel flavor, which comprises the following steps: (1) taking cane molasses, and diluting; (2) performing biological fermentation, inactivating, cooling and filtering; (3) performing chromatography through four macroporous resin columns connected in series, performing gradient elution, and performing low-temperature concentration; and (4) carrying out alcohol precipitation, concentration, paste preparation and drying to obtain the sugarcane polyphenol composition. According to the method, biological fermentation and gradient elution of different solvents in which four macroporous resins are connected in series are applied, separation of effective components and impurities is utilized, the purity of the sugarcane polyphenol is optimized, the process is simple, the whole process is safe, the method is suitable for large-scale factory production, the polyphenol content in the obtained sugarcane polyphenol composition is larger than 20%, and the composition is good in water solubility, has scorch aroma and is suitable for industrial production. The method can be widely applied to food, health-care products, beverages and the like.

Description

Technical Field

[0001] The invention relates to the technical field of natural active substance extraction, and in particular to a method for preparing sugarcane polyphenols with caramel flavor. Background Art

[0002] Every year, my country produces a huge amount of sugarcane, and the sugarcane resources are extremely rich. Sugarcane contains phenolic acids, flavonoids and other phenolic compounds. The polyphenol extracts in sugarcane have certain effects on scavenging free radicals and anti-oxidation. It can also be used to reduce cholesterol, fight obesity, fight diabetes, fight bacteria, and preserve fresh food. Sugarcane molasses is the thick liquid left after the sugar industry squeezes out the sugarcane juice through heating, neutralization, precipitation, filtration, concentration, crystallization and other processes. Molasses is not only an energy raw material, but also has the advantages of fast digestion and absorption, improved palatability, reduced dust, and improved particle quality. Sugarcane molasses is rich in a large number of high-value-added products, so it is necessary to increase the research on sugarcane molasses, innovate simpler and more efficient extraction methods to extract natural active substances in sugarcane molasses, and develop a broader research and utilization field for sugarcane polyphenol extracts.

[0003] Currently, most of the process research is focused on sugarcane peel and sugarcane scum. For example, CN105816702A discloses a method for preparing a sugarcane polyphenol extract, using sugarcane or bagasse as raw materials. There is little research on molasses after sugar extraction. The present invention provides a preparation scheme for the research and development of obtaining sugarcane polyphenols from molasses. Summary of the invention

[0004] The object of the present invention is to provide a method for preparing a sugarcane polyphenol composition with caramel flavor by using sugarcane molasses, wherein the method separates effective components from impurities and optimizes the purity of the sugarcane polyphenol.

[0005] The preparation method comprises the following steps:

[0006] (1) Take sugarcane molasses and dilute it;

[0007] (2) Biological fermentation, inactivation, cooling, and filtration;

[0008] (3) chromatography through four large-pore columns connected in series, gradient elution, and cryogenic concentration;

[0009] (4) alcohol precipitation, concentration, paste making, and drying to obtain a sugarcane polyphenol composition.

[0010] In step (1), the polyphenol content of the sugarcane molasses is 2.5% and the solid content is 70%, and pure water is added to dilute the solid content to 7-10%.

[0011] Step (2) specifically comprises adding highly active yeast, fermenting at 30-32° C. for 4-6 hours, then heating to greater than 98° C., maintaining for 5-10 minutes, cooling to 25-30° C., filtering through a 200-mesh filter cloth, and obtaining a medicinal solution 1.

[0012] The highly active yeast is Angel Yeast, and the added amount is 0.5-1wt.% of the amount of sugarcane molasses.

[0013] The model of the macroporous resin is xda-6, DM18 or LK-83S, and the manufacturer is Xi'an Lanxiao Biotechnology Co., Ltd.

[0014] The mass ratio of the amount of each macroporous resin to the solid content in the sugarcane molasses is 1:1.

[0015] The step (3) of passing through four series-connected macroporous columns for chromatography comprises: drug solution 1 first enters the first resin, and after entering, it is eluted with pure water, and the effluent 1 and the water washing liquid 1 are collected and combined into drug solution 2; drug solution 2 enters the second resin, and after entering, it is eluted with pure water, and the effluent 2 and the water washing liquid 2 are collected and combined into drug solution 3; drug solution 3 enters the third resin, and after entering, it is eluted with pure water, and the effluent 3 and the water washing liquid 3 are collected and combined into drug solution 4; drug solution 4 enters the fourth resin, and after entering, the four resins are eluted again with pure water in turn, until the effluent is clear and transparent; the amount of pure water used for elution after entering the first to third resins is 0.5-1 times the amount of resin, and the amount of pure water used for elution again after entering the fourth resin is 2-4 times the amount of resin.

[0016] The gradient elution described in step (3) includes: eluting four resins simultaneously, first eluting with 10% ethanol to wash away impurities, and then eluting with 50-60% ethanol. After the effluent is dried and the polyphenol content is detected to be greater than 0.8%, the ethanol eluate is collected and the eluates of the second and third resins are combined.

[0017] The concentration in step (3) is to concentrate the combined eluate using a low-temperature concentrator at a temperature < 60° C. until it is alcohol-free and the concentration is 48-52 BRIX.

[0018] Furthermore, the elution volume of the 10% ethanol elution is 2-5 times the amount of the resin, and the elution flow rate is 1-1.5 BV / h; the elution volume of the 50-60% ethanol elution is 2-4 times the amount, and the elution flow rate is 0.8-2 BV / h.

[0019] Step (4) specifically comprises adding 3-4 times the volume of pure methanol or anhydrous ethanol of the concentrate, sealing well, keeping stirring constantly, keeping at 4-6° C. for 1-2 hours, centrifuging, taking the supernatant and concentrating it to be alcohol-free, making a paste to a brix of 35-40 BRIX, spray drying, and obtaining a sugarcane polyphenol composition.

[0020] The sugarcane polyphenol composition contains 20-25% of polyphenols, 1-20% of flavonoids and 0.1-2% of polysaccharides.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) Biofermentation plus gradient elution of four macroporous resins in series with different solvents, utilizing the separation of active ingredients and impurities, was used to optimize the purity of sugarcane polyphenols, and the polyphenol content obtained was greater than 20%.

[0023] (2) Low temperature stirring and alcohol precipitation further increase the content of polyphenols and effective ingredients, while also taking into account the decolorization effect. At the same time, water is added for concentration, ethanol residue is removed, and spraying is performed to ensure that the product has no solvent residue and meets the needs of the development of big health.

[0024] (3) The preparation method is simple in process, safe throughout the process, environmentally friendly, and suitable for large-scale factory production. The obtained sugarcane polyphenol composition has good water solubility and a caramel flavor, and can be widely used in food, health products, beverages, etc. DETAILED DESCRIPTION

[0025] The specific implementation of the present invention is further described below in conjunction with examples, but the implementation and protection of the present invention are not limited thereto. It should be noted that if there are processes that are not particularly described in detail below, they can be implemented or understood by those skilled in the art with reference to the prior art.

[0026] Example 1

[0027] Take 428.6 g of sugar cane molasses (polyphenol content: 2.5%, solid content: 70%) and dilute it with pure water until the solid content is 7%.

[0028] Add 3g of highly active yeast (Angel Yeast), ferment at 32℃ for 5 hours, heat to 99℃, keep for 10 minutes to inactivate, cool the solution, wait for the solution to cool to 28℃, and filter it through a 200-mesh filter cloth.

[0029] The drug solution was chromatographed through 4 macroporous resins (DM18, Xi'an Lanxiao Biotechnology Co., Ltd.), with each resin using 300 g. The drug solution first entered the first resin (the column flow rate was 1 BV / h), and after it was completed, it was eluted with 300 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the second resin, and after the second resin was completely filled with the drug solution, it was eluted with 300 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the third resin, and after the third resin was completely filled with the drug solution, it was eluted with 300 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the fourth resin, and after the fourth resin was completely filled with the drug solution, the four resins were eluted again with pure water in turn until the effluent was clear and transparent (the dosage was 1200 mL of pure water). Gradient elution, four resins are eluted at the same time, first eluted with 10% ethanol, the elution volume is 600mL, the elution flow rate is 1.5BV / h, wash away impurities; then eluted with 50% ethanol, the elution volume is 900mL, the elution flow rate is 1.5BV / h; after the collected effluent is dried and the polyphenol content is greater than 0.8%, start collecting 50% ethanol eluate, and combine the eluate of the 2nd and 3rd resins. Use a low-temperature concentrator (temperature <60℃) to concentrate to alcohol-free, and concentrate to 48BRIX. Add 3 times the volume of anhydrous ethanol, seal well, keep stirring continuously, keep at 6℃ for 1 hour, centrifuge, take the supernatant and concentrate to alcohol-free, and make a paste to 35BRIX.

[0030] The sugarcane polyphenol composition was obtained by spray drying. The composition was found to contain 22% polyphenols, 20% flavonoids and 2% polysaccharides.

[0031] Example 2

[0032] Take 428.6 g of sugar cane molasses (polyphenol content: 2.5%, solid content: 70%) and dilute it with pure water until the solid content is 8%.

[0033] Add 1.5g of highly active yeast (Angel Yeast), ferment at 30℃ for 4 hours, heat to 100℃, keep for 10 minutes to inactivate, cool the solution, wait for the solution to cool to 25℃, and filter it through a 200-mesh filter cloth.

[0034] The drug solution was chromatographed through 4 macroporous resins (XDA-6, Xi'an Lanxiao Biotechnology Co., Ltd.) connected in series, with each resin using 300 g. The drug solution first entered the first resin (the column flow rate was 1.5 BV / h), and after it was completed, it was eluted with 150 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the second resin, and after the second resin was completely filled with the drug solution, it was eluted with 150 mL of pure water, and the effluent and the washing liquid were collected and combined together; then it entered the third resin, and after the third resin was completely filled with the drug solution, it was eluted with 150 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the fourth resin, and after the fourth resin was completely filled with the drug solution, the four resins were eluted again with pure water in turn, and the effluent was eluted until the effluent was clear and transparent (the dosage was 900 mL of pure water). Gradient elution, four resins are eluted at the same time, first eluted with 10% ethanol, the elution volume is 900mL, the elution flow rate is 1BV / h, and impurities are washed away; then eluted with 60% ethanol, the elution volume is 900mL, and the elution flow rate is 0.8BV / h; after the collected effluent is dried and the polyphenol content is detected to be greater than 0.8%, start collecting 60% ethanol eluate and combine the eluate of the 2nd and 3rd resins. Use a low-temperature concentrator (temperature <60℃) to concentrate to no alcohol and concentrate to 52BRIX. Add 4 times the volume of pure methanol, seal well, keep stirring continuously, keep at 4℃ for 2 hours, centrifuge, take the supernatant and concentrate to no alcohol, and make a paste to 40BRIX.

[0035] The sugarcane polyphenol composition was obtained by spray drying. The composition was found to contain 20% polyphenols, 15% flavonoids and 1% polysaccharides by testing.

[0036] Example 3

[0037] Take 428.6 g of sugar cane molasses (polyphenol content: 2.5%, solid content: 70%) and dilute it with pure water until the solid content is 10%.

[0038] Add 1.5g of highly active yeast (Angel Yeast), ferment at 31°C for 6 hours, heat to 99°C, keep for 8 minutes to inactivate, cool the solution, wait for the solution to cool to 28°C, and filter it through a 200-mesh filter cloth.

[0039] The drug solution was chromatographed through 4 macroporous resins (LK-83S, Xi'an Lanxiao Biopharmaceutical Co., Ltd.) connected in series, with each resin using 300 g. The drug solution first entered the first resin (column flow rate was 1.5 BV / h), and after it was completed, it was eluted with 300 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the second resin, and after the second resin was completely filled with the drug solution, it was eluted with 300 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the third resin, and after the third resin was completely filled with the drug solution, it was eluted with 300 mL of pure water, and the effluent and the washing liquid were collected and combined together; it entered the fourth resin, and after the fourth resin was completely filled with the drug solution, the four resins were eluted again with pure water in turn until the effluent was clear and transparent (the dosage was 600 mL of pure water). Gradient elution, four resins are eluted at the same time, first eluted with 10% ethanol, the elution volume is 1500mL, the elution flow rate is 1BV / h, and impurities are washed away; then eluted with 55% ethanol, the elution volume is 900mL, and the elution flow rate is 2BV / h; after the collected effluent is dried and the polyphenol content is detected to be greater than 0.8%, start collecting 55% ethanol eluate, and combine the eluate of the 2nd and 3rd resins. Use a low-temperature concentrator (temperature <60℃) to concentrate to alcohol-free and concentrate to 50BRIX. Add 4 times the volume of pure methanol, seal well, keep stirring continuously, keep at 5℃ for 1.5 hours, centrifuge, take the supernatant and concentrate to alcohol-free, and make a paste to 38BRIX.

[0040] The sugarcane polyphenol composition was obtained by spray drying. The composition was found to contain 22% polyphenols, 18% flavonoids and 0.5% polysaccharides.

[0041] The above embodiments are only preferred implementation modes of the present invention and are only used to explain the present invention rather than to limit the present invention. Any changes, substitutions, modifications, etc. made by those skilled in the art without departing from the spirit of the present invention should fall within the scope of protection of the present invention.

Claims

1. A method for preparing sugarcane polyphenols with caramel flavor, characterized in that: The preparation method comprises the following steps: (1) Take sugarcane molasses and dilute it; (2) Biological fermentation, inactivation, cooling, and filtration; (3) chromatography through four macroporous resin columns connected in series, gradient elution, and cryogenic concentration; (4) alcohol precipitation, concentration, paste making, and drying to obtain a sugarcane polyphenol composition.

2. The preparation method according to claim 1, characterized in that: In step (1), the polyphenol content of the sugarcane molasses is 2.5% and the solid content is 70%, and pure water is added to dilute the solid content to 7-10%.

3. The preparation method according to claim 1, characterized in that: Step (2) specifically comprises adding highly active yeast, fermenting at 30-32° C. for 4-6 hours, then heating to greater than 98° C., maintaining for 5-10 minutes, cooling to 25-30° C., filtering through a 200-mesh filter cloth, and obtaining liquid 1; the highly active yeast is Angel Yeast, and the amount added is 0.5-1wt.% of the amount of sugarcane molasses.

4. The preparation method according to claim 1, characterized in that: The mass ratio of the amount of each macroporous resin to the solid content in the sugarcane molasses is 1:

1. The model of the macroporous resin is xda-6, DM18 or LK-83S, and the manufacturer is Xi'an Lanxiao Biomedicine Co., Ltd.

5. The preparation method according to claim 1, characterized in that: The step (3) of passing through four series-connected macroporous columns for chromatography comprises: drug solution 1 first enters the first resin, and after entering, it is eluted with pure water, and the effluent 1 and the water washing liquid 1 are collected and combined into drug solution 2; drug solution 2 enters the second resin, and after entering, it is eluted with pure water, and the effluent 2 and the water washing liquid 2 are collected and combined into drug solution 3; drug solution 3 enters the third resin, and after entering, it is eluted with pure water, and the effluent 3 and the water washing liquid 3 are collected and combined into drug solution 4; drug solution 4 enters the fourth resin, and after entering, the four resins are eluted again with pure water in turn, until the effluent is clear and transparent; the amount of pure water used for elution after entering the first to third resins is 0.5-1 times the amount of resin, and the amount of pure water used for elution again after entering the fourth resin is 2-4 times the amount of resin.

6. The preparation method according to claim 1, characterized in that: The gradient elution described in step (3) includes: eluting four resins simultaneously, first eluting with 10% ethanol to wash away impurities, and then eluting with 50-60% ethanol. After the effluent is dried and the polyphenol content is detected to be greater than 0.8%, the ethanol eluate is collected and the eluates of the second and third resins are combined.

7. The preparation method according to claim 1, characterized in that: The concentration in step (3) is to concentrate the combined eluate using a low-temperature concentrator at a temperature < 60° C. until it is alcohol-free and the concentration is 48-52 BRIX.

8. The preparation method according to claim 6, characterized in that: The elution volume of the 10% ethanol elution is 2-5 times the amount of the resin, and the elution flow rate is 1-1.5 BV / h; the elution volume of the 50-60% ethanol elution is 2-4 times the amount, and the elution flow rate is 0.8-2 BV / h.

9. The preparation method according to claim 1, characterized in that: Step (4) specifically comprises adding 3-4 times the volume of pure methanol or anhydrous ethanol of the concentrate, sealing well, keeping stirring constantly, keeping at 4-6° C. for 1-2 hours, centrifuging, taking the supernatant and concentrating it to be alcohol-free, making a paste to a brix of 35-40 BRIX, spray drying, and obtaining a sugarcane polyphenol composition.

10. The preparation method according to claim 1, characterized in that: The sugarcane polyphenol composition contains 20-25% polyphenol, 1-20% flavonoids and 0.1-2% polysaccharide.