阅读说明：本技术 一种黑果腺肋花楸多酚提取工艺 (A kind of Black Box Tracing polyphenol extraction technology ) 是由 王让成 韩泽民 刘红卫 贾慧 朱日超 于 2019-08-22 设计创作，主要内容包括：本发明涉及一种黑果腺肋花楸多酚提取工艺,其包括以下提取步骤：S1：对黑果腺肋花楸果渣进行预处理,制得含有黑果腺肋花楸多酚的原料液；S2：采用双水相萃取原料液的活性成分,并采用微波辅助提取法,制得黑果腺肋花楸多酚粗提液；微波功率300-500W、提取时间5-10min；S3：将黑果腺肋花楸多酚粗提液旋蒸除醇后,制得A混合物；S4：在A混合物中加入稳定剂,搅拌均匀后,制得黑果腺肋花楸多酚提取液；S5：将黑果腺肋花楸多酚提取液冷冻干燥处理得到黑果腺肋花楸多酚提取物。通过选用黑果腺肋花楸果渣作为原料,对其进行预处理,便于之后的活性物质提取,且减少资源的浪费,提高资源的利用率。(The present invention relates to a kind of Black Box Tracing polyphenol extraction technologies comprising following extraction step: S1: pre-processing Black Box Tracing pomace, and the material liquid containing Black Box Tracing polyphenol is made；S2: using the active constituent of aqueous two-phase extraction material liquid, and microwave―assisted extraction is used, Black Box Tracing polyphenolic extract is made；Microwave power 300-500W, extraction time 5-10min；S3: after Black Box Tracing polyphenolic extract revolving is removed alcohol, A mixture is made；S4: being added stabilizer in A mixture, after mixing evenly, Black Box Tracing polyphenol extracting solution is made；S5: Black Box Tracing polyphenol extracting solution freeze-drying process is obtained into Black Box Tracing polyphenol extract.By selecting Black Box Tracing pomace as raw material, it is pre-processed, is extracted convenient for active material later, and reduces the waste of resource, improves the utilization rate of resource.)

1. a kind of Black Box Tracing polyphenol extraction technology, it is characterised in that: including following extraction step:

S1: pre-processing Black Box Tracing pomace, and the material liquid containing Black Box Tracing polyphenol is made；

S2: using the active constituent of aqueous two-phase extraction material liquid, and microwave―assisted extraction is used, it is more that Black Box Tracing is made Phenol crude extract；Microwave power 300-500W, extraction time 5-10min；

S3: after Black Box Tracing polyphenolic extract revolving is removed alcohol, A mixture is made；

S4: being added stabilizer in A mixture, after mixing evenly, Black Box Tracing polyphenol extracting solution is made；

S5: Black Box Tracing polyphenol extracting solution freeze-drying process is obtained into Black Box Tracing polyphenol extract.

2. a kind of Black Box Tracing polyphenol extraction technology according to claim 1, it is characterised in that: the S1 pretreatment Including following preparation process:

1) ethyl alcohol for being 40-60% by Black Box Tracing pomace and mass concentration, after solid-liquid ratio 1:(20-30) g/mL mixing, Enzyme auxiliary agent is added and is heated to 50-60 DEG C, insulation reaction 40-60min after mixing evenly, is then assisted using ultrasonic wave After reacting 20-30min, the first mixed liquor is made in method；

2) in the first mixed liquor,-two -2- ethylhexyl sodium sulfonate of succinic acid and n-hexane is added, after stirring 30-50min, In It is evaporated under reduced pressure at 30-35 DEG C, recycles n-hexane, residual mixed liquor is the second mixed liquor；

3) in the second mixed liquor, the first auxiliary agent is added, and be warming up to 40-50 DEG C, after mixing evenly, using being filtered under diminished pressure, obtains The clear liquid arrived is the material liquid containing Black Box Tracing polyphenol；First auxiliary agent and the Black Box Tracing pomace Weight ratio is (1-3): 1.

3. a kind of Black Box Tracing polyphenol extraction technology according to claim 2, it is characterised in that: the enzyme auxiliary agent with The weight ratio of the Black Box Tracing pomace is (0.03-0.07): 1.

4. a kind of Black Box Tracing polyphenol extraction technology according to claim 2, it is characterised in that: the enzyme auxiliary agent packet Include following weight percent meter raw material: cellulase 20-40%, pectinesterase 10-30%, disodium hydrogen phosphate 1-5%, biphosphate Sodium 3-7% and excess water.

5. a kind of Black Box Tracing polyphenol extraction technology according to claim 2, it is characterised in that: the succinic acid- The weight ratio of two -2- ethylhexyl sodium sulfonates and the Black Box Tracing pomace is (0.01-0.05): 1, the n-hexane with The volume ratio of first mixed liquor is (0.8-1.2): 1.

6. a kind of Black Box Tracing polyphenol extraction technology according to claim 2, it is characterised in that: first auxiliary agent Including following weight percent agent raw material: lysine 10-14%, histidine 16-20%, ethyl alcohol 30-34% and excess water；Made Concentration of alcohol is identical as concentration of alcohol used in the step 1.

7. a kind of Black Box Tracing polyphenol extraction technology according to claim 2, it is characterised in that: double water in the S2 Mutually extraction comprises the following steps that: ethyl alcohol and ammonium sulfate are added into material liquid, it is lasting to stir, and pH adjusting agent is added, it adjusts Section to pH value is 2.5-3.5, stands after mixing evenly, phase alcohol liquid i.e. Black Box Tracing polyphenolic extract is taken after layering.

8. a kind of Black Box Tracing polyphenol extraction technology according to claim 7, it is characterised in that: made in the S2 Concentration of alcohol is identical as concentration of alcohol used in the S1, and the solid-liquid ratio of the ethyl alcohol and Black Box Tracing pomace For 1:(30-50) g/mL；The ammonium sulfate additional amount and the weight ratio of Black Box Tracing pomace are (0.04-0.08): 1.

9. a kind of Black Box Tracing polyphenol extraction technology according to claim 7, it is characterised in that: the pH adjusting agent Including following weight percent meter raw material: glacial acetic acid 30-40%, aspartic acid 16-20%, glutamic acid 10-14% and excess water.

10. a kind of Black Box Tracing polyphenol extraction technology according to claim 1, it is characterised in that: the stabilizer Weight ratio with Black Box Tracing pomace is (0.02-0.06): 1；The stabilizer includes following weight percent meter raw material: Potassium sorbate 0.6-1%, histidine 8-12%, isoleucine 6-10%, tryptophan 10-14% and excess water.

Technical field

The present invention relates to the technical fields that natural plant polyphenols extract, and mention more particularly, to a kind of Black Box Tracing polyphenol Taking technique.

Background technique

Black Box Tracing is a kind of small berries tree species with very high nutritive value and medical value, is rich in fruit Polyphenol substance be even more there are the physiological activity such as cardiovascular disease resistant, antitumor, anti-oxidant, have very high Development volue.

Due to its high nutritive value, pole was liked by people in recent years, by-product jam, fruit juice and processing wine It is even more very popular.

In process, the pomace that Black Box Tracing generates generally all directly is filled out as fertilizer due to bad processing Bury in soil.But many nutritional ingredients are still remained in the pomace of Black Box Tracing in fact, directly landfill will cause The nutrition of this part wastes.

Summary of the invention

The object of the present invention is to provide a kind of Black Box Tracing polyphenol extraction technologies, by selecting Black Box Tracing fruit Slag pre-processes it as raw material, extracts convenient for active material later, and reduces the waste of resource, improves resource Utilization rate.

Foregoing invention purpose of the invention has the technical scheme that

A kind of Black Box Tracing polyphenol extraction technology, including following extraction step:

S1: pre-processing Black Box Tracing pomace, and the material liquid containing Black Box Tracing polyphenol is made；

S2: using the active constituent of aqueous two-phase extraction material liquid, and using microwave―assisted extraction, and black fruit gland rib flower is made Chinese catalpa polyphenolic extract；Microwave power 300-500W, extraction time 5-10min；

S3: after Black Box Tracing polyphenolic extract revolving is removed alcohol, A mixture is made；

S4: being added stabilizer in A mixture, after mixing evenly, Black Box Tracing polyphenol extracting solution is made；

S5: Black Box Tracing polyphenol extracting solution freeze-drying process is obtained into Black Box Tracing polyphenol extract.

By using above-mentioned technical proposal, in S1, by being pre-processed to Black Box Tracing pomace, convenient for later Active material extracts, and improves Black Box Tracing polyphenol dissolution rate.

In S2, S3, to extract Black Box Tracing polyphenol.

In S4, there is stabilizer by being added, improve the stability of Black Box Tracing polyphenol, it is more to increase Black Box Tracing The holding time of phenol and using effect.

The present invention is further arranged to: the S1 pretreatment includes following preparation process:

1) ethyl alcohol for being 40-60% by Black Box Tracing pomace and mass concentration, it is mixed by solid-liquid ratio 1:(20-30) g/mL After conjunction, enzyme auxiliary agent is added and is heated to 50-60 DEG C, insulation reaction 40-60min after mixing evenly, then uses ultrasonic wave After reacting 20-30min, the first mixed liquor is made in auxiliary law；

2) in the first mixed liquor,-two -2- ethylhexyl sodium sulfonate of succinic acid and n-hexane is added, stirs 30-50min Afterwards, it is evaporated under reduced pressure at 30-35 DEG C, recycles n-hexane, residual mixed liquor is the second mixed liquor；

3) in the second mixed liquor, the first auxiliary agent is added, and is warming up to 40-50 DEG C, after mixing evenly, using depressurizing Filter, obtained clear liquid are the material liquid containing Black Box Tracing polyphenol；First auxiliary agent and the Black Box Tracing The weight ratio of pomace is (1-3): 1.

By using above-mentioned technical proposal, due to the Black Box Tracing polyphenol in Black Box Tracing pomace content Through seldom, remain in Black Box Tracing polyphenol in Black Box Tracing pomace generally all in pericarp.Due in pericarp Cell wall is generally all more firm, not easily passs through conventional means and extracts Black Box Tracing polyphenol.

In step 1, there is enzyme auxiliary agent by being added, be hydrolyzed to the cell wall to plant, so that cell wall It is not located at intracellular remaining Black Box Tracing polyphenol in protection, so that the Black Box Tracing polyphenol remained in cell is easy In ethanol in dissolution, it is extracted convenient for Black Box Tracing polyphenol later.

Since Black Box Tracing polyphenol is generally all incorporated on protein, by increasing the dissolution of protein in water Amount, can be improved the meltage of Black Box Tracing polyphenol in water, to improve the extracted amount of Black Box Tracing polyphenol

In step 2, there are-two -2- ethylhexyl sodium sulfonate of succinic acid and n-hexane by being added, in A mixed liquor Form reverse micelle, to extract protein and Black Box Tracing polyphenol in aqueous solution so that protein in aqueous solution and Black Box Tracing polyphenol content is reduced, so that the protein and Black Box Tracing polyphenol that are located in cell are precipitated, is mentioned The content of high Black Box Tracing polyphenol.

Ethyl alcohol in step 1 can be used as cosurfactant in step 2, improve the active and steady of biocatalytic reaction It is qualitative.

Since the boiling point of n-hexane is at 68 DEG C, and Black Box Tracing polyphenol can decompose at relatively high temperatures, by using Vacuum distillation, so that Black Box Tracing polyphenol will not be made to decompose under the purpose for realizing separation n-hexane.At 30-35 DEG C Lower vacuum distillation improves the speed of vacuum distillation, the time required to reducing.

Addition has the first auxiliary agent, breaks the balance of reverse micelle extraction, so that the protein being located in reverse micelle comes out, is convenient for Black Box Tracing polyphenol is extracted later.

The present invention is further arranged to: the weight ratio of the enzyme auxiliary agent and the Black Box Tracing pomace is (0.03- 0.07):1。

By using above-mentioned technical proposal, prevent enzyme auxiliary agent excessive, so that in the process of Research Advances in Reversed Micellar Extraction In, excessive enzyme auxiliary agent enters in reverse micelle, to influence with Black Box Tracing polyphenolic protein and black fruit gland rib flower The extraction of Chinese catalpa polyphenol.Enzyme auxiliary agent is very few, and the cell wall amount that will lead to hydrolysis is reduced, and influences the precipitation of Black Box Tracing polyphenol Amount.

The present invention is further arranged to: the enzyme auxiliary agent includes following weight percent meter raw material: cellulase 20- 40%, pectinesterase 10-30%, disodium hydrogen phosphate 1-5%, sodium dihydrogen phosphate 3-7% and excess water.

By using above-mentioned technical proposal, plant cell wall is mainly cellulose and pectin, and used addition has cellulase And pectinesterase, to hydrolyzing plant cell wall.

Due to not only containing Black Box Tracing polyphenol into the cell, also contain remaining nutriment, some nutriments In acid or alkalinity, the pH value of solution will affect, to influence the activity of cellulase and pectinesterase, reduce cellulase With pectinesterase to the hydrolysis rate of plant cell wall.

There are disodium hydrogen phosphate and sodium dihydrogen phosphate by being added, to adjust solution ph, the pH value of solution is maintained to tend to Stable state reduces solution ph to the activity influence of cellulase and pectinesterase.

In Research Advances in Reversed Micellar Extraction, the pH value of solution has very big shadow to the rate of extraction and extraction quantity of protein It rings, has the addition of disodium hydrogen phosphate and sodium dihydrogen phosphate by being added, can automatically adjust required when Research Advances in Reversed Micellar Extraction PH value, be convenient for worker operation, improve the operability of worker.And the amount of Research Advances in Reversed Micellar Extraction is improved, to improve black The content of fruit gland rib Sorbus alnifloria polyphenol

The present invention is further arranged to:-two -2- ethylhexyl sodium sulfonate of succinic acid and the Black Box Tracing fruit The weight ratio of slag is (0.01-0.05): 1, the volume ratio of the n-hexane and first mixed liquor is (0.8-1.2): 1.

By using above-mentioned technical proposal, when the addition excess of-two -2- ethylhexyl sodium sulfonate of succinic acid and n-hexane, The stability that will affect Black Box Tracing polyphenol makes Black Box Tracing polyphenol be easy to fail.

When the addition of-two -2- ethylhexyl sodium sulfonate of succinic acid and n-hexane is very few, Black Box Tracing polyphenol can be reduced Extracted amount.

The present invention is further arranged to: first auxiliary agent includes following weight percent agent raw material: lysine 10- 14%, histidine 16-20%, ethyl alcohol 30-34% and excess water；Used in used concentration of alcohol and the step 1 Concentration of alcohol is identical.

By using above-mentioned technical proposal, there are lysine and histidine by being added, to adjust the pH value of solution, so that The pH value of solution is in alkalinity, and the protein convenient for being located in reverse micelle is precipitated, to improve Black Box Tracing polyphenol in solution Content.

Since alkaline environment is easy to influence the stability of Black Box Tracing polyphenol, by using lysine and histidine, All it is one of element necessary to human body and plant since the two is all basic amino acid, is a kind of substance of safe green, it is right The influence of Black Box Tracing polyphenol is smaller, improves the stability of Black Box Tracing polyphenol.

And histidine can not only be used for proton donor, but also as proton acceptor, so that the pH value of solution tends to balance, It prevents the pH value of the solution of solution too high or too low, influences the stability of Black Box Tracing polyphenol.

Ethyl alcohol is as cosurfactant, and the protein convenient for being located in reverse micelle is precipitated, to improve black fruit in solution The content of gland rib Sorbus alnifloria polyphenol.Using the ethyl alcohol of same concentration, worker need to only deploy a kind of concentration ethanol solution, grasp convenient for worker Make, improve work task efficiency, realizes production on a large scale.

The present invention is further arranged to: aqueous two-phase extraction comprises the following steps that in the S2: being added into material liquid Ethyl alcohol and ammonium sulfate, it is lasting to stir, and pH adjusting agent is added, being adjusted to pH value is 2.5-3.5, is stood after mixing evenly, is layered After take phase alcohol liquid i.e. Black Box Tracing polyphenolic extract.

The present invention is further arranged to: concentration of alcohol used in the S2 and concentration of alcohol phase used in the S1 Together, and the ethyl alcohol and the solid-liquid ratio of Black Box Tracing pomace are 1:(30-50) g/mL；The ammonium sulfate additional amount and black fruit The weight ratio of gland rib Sorbus alnifloria pomace is (0.04-0.08): 1.

By using above-mentioned technical proposal, aqueous two-phase extraction uses ethyl alcohol and ammonium sulfate for extractant, will not introduce new Element improves the degree of purity of Black Box Tracing polyphenolic extract.

When using aqueous two-phase extraction feed liquid, the amount between ethyl alcohol and ammonium sulfate is excessive or very few all will affect than critically important The extraction of required object, and the amount ratio between the required ethyl alcohol and ammonium sulfate of different extracts is all different, using such range The extraction that interior ethyl alcohol and volume of ammonium sulfate compare Black Box Tracing polyphenol is suitble to the most, improves Black Box Tracing polyphenol and extracts While amount, Black Box Tracing polyphenol will not be made to fail.

The present invention is further arranged to: the pH adjusting agent includes following weight percent meter raw material: glacial acetic acid 30- 40%, aspartic acid 16-20%, glutamic acid 10-14% and excess water.

By using above-mentioned technical proposal, glacial acetic acid and esterification can occur with ethyl alcohol, generate ethyl acetate, acetic acid second The stability and solubility of Black Box Tracing polyphenol can be improved in ester.

Both aspartic acid and glutamic acid are all acidic amino acids, are all one of elements necessary to human body and plant, are A kind of substance of safe green, the influence to Black Box Tracing polyphenol is smaller, and Black Box Tracing polyphenol can be improved Stability and meltage.

The present invention is further arranged to: the weight ratio of the stabilizer and Black Box Tracing pomace is (0.02-0.06): 1；The stabilizer includes following weight percent meter raw material: potassium sorbate 0.6-1%, histidine 8-12%, isoleucine 6- 10%, tryptophan 10-14% and excess water.

By using above-mentioned technical proposal, potassium sorbate prevents Black Box Tracing polyphenol by micro- life as preservative The influence of object and decompose, improve Black Box Tracing polyphenol using effect and storage time.

Histidine can not only be used for proton donor, but also as proton acceptor, so that the pH value of solution tends to balance, prevent Only the pH value of the solution of solution is too high or too low, influences the stability of Black Box Tracing polyphenol.

Isoleucine can decompose generation glucose, so that water activity gradually decreases in solution, to reduce black fruit The degradation of gland rib Sorbus alnifloria polyphenol.

Ultraviolet light can be absorbed in tryptophan, prevents Black Box Tracing polyphenol by fast degradation after illumination, influences its Using effect and storage time.

Histidine, isoleucine and tryptophan three are all one of neutral amino acid, element necessary to human body and plant, It is a kind of substance of safe green, the influence to Black Box Tracing polyphenol is smaller.And when edible, the body of people will not influence Body health can also improve human body and absorb to Black Box Tracing polyphenol.

In conclusion advantageous effects of the invention are as follows:

1, by selecting Black Box Tracing pomace as raw material, it is pre-processed, convenient for active material later It extracts, and reduces the waste of resource, improve the utilization rate of resource；

2, there are-two -2- ethylhexyl sodium sulfonate of succinic acid and n-hexane by being added, using Research Advances in Reversed Micellar Extraction, To improve the amount of dissolution of Black Box Tracing polyphenol；

3, there is stabilizer by being added, improve the stability of Black Box Tracing polyphenol, increase Black Box Tracing polyphenol Holding time and using effect.

Specific embodiment