阅读说明：本技术 接骨木莓压片糖果及接骨木莓提取物的制备方法 (Preparation method of elderberry tablet candy and elderberry extract ) 是由 赵海瑛 张艳立 冯亚净 张香香 谢勇 程坤杰 于 2020-12-17 设计创作，主要内容包括：本发明公开了一种接骨木莓压片糖果及接骨木莓提取物的制备方法。制备方法为：将经挑选的接骨木莓打浆后,加入纤维素酶和果胶酶进行酶解,之后接种酵母菌,发酵后在超声处理下进行提取得到发酵初液,泵入发酵罐,接种醋酸菌,发酵到一定条件后,再次超声提取,滤液经旋转蒸发后冻干成粉。本发明所得提取物中花青素含量高达77.2-81.3mg/g。本发明整个制备过程未使用有机提取溶媒,无溶剂残留等食品安全风险,与传统提取方式比较具有设备简单,控制条件温和,操作简便,针对花青素提取效率和提取特异性高,经济环保,该方法有效降低生产成本,利于产业化规模化生产。本发明还提供了一种添加接骨木莓提取物的压片糖果。(The invention discloses a method for preparing elderberry tablet candy and an elderberry extract. The preparation method comprises the following steps: pulping selected elderberry, adding cellulase and pectinase for enzymolysis, inoculating saccharomycetes, fermenting, extracting under ultrasonic treatment to obtain primary fermentation liquid, pumping into a fermentation tank, inoculating acetic acid bacteria, fermenting to a certain condition, performing ultrasonic extraction again, performing rotary evaporation on the filtrate, and freeze-drying to obtain powder. The anthocyanin content in the extract obtained by the invention is up to 77.2-81.3 mg/g. The whole preparation process of the invention does not use organic extraction solvent, has no food safety risks such as solvent residue and the like, has simple equipment, mild control condition, simple and convenient operation compared with the traditional extraction mode, has high extraction efficiency and extraction specificity aiming at anthocyanin, is economic and environment-friendly, effectively reduces the production cost, and is beneficial to industrialized large-scale production. The invention also provides a tablet candy added with the elderberry extract.)

1. The bonesetting raspberry tabletting candy is characterized by comprising the following components in parts by weight: 0.2-0.4 part of elderberry extract, 0.5-0.8 part of acerola cherry concentrated powder, 0.05-0.08 part of vitamin C, 7-9 parts of sorbitol, 0.1-0.3 part of citric acid, 0.005-0.007 part of aspartame, 0.2-0.3 part of magnesium stearate and 0.1-0.2 part of edible essence.

2. The preparation method of the elderberry extract is characterized by comprising the following steps:

a. crushing elderberry into pulp, and adding cellulase and pectinase for enzymolysis;

b. inoculating 2-5wt% of yeast after enzymolysis, fermenting at 18-23 deg.C in dark place, stopping fermentation until alcoholic strength is 5.5-8.7 °, subjecting the fermentation product to ultrasonic extraction at 40-45 deg.C for 1-2 hr, and filtering to obtain primary fermentation solution;

c. pumping the primary fermentation liquid into a fermentation tank, adding 8-10 wt% of acetic acid bacteria, keeping the ventilation amount of sterile air at 0.06-0.08vvm, keeping the fermentation temperature at 28-33 ℃, stopping fermentation when the pH value is 2.8-3 after stirring and fermenting, placing the fermentation product at 40-45 ℃ for ultrasonic extraction for 1-2h, and filtering;

d. and performing rotary evaporation on the filtrate, and then freeze-drying into powder to obtain the elderberry extract.

3. The preparation method of claim 2, wherein in step a, the cellulase is 0.5-1wt%, the pectinase is 0.07-0.1wt%, the enzymolysis temperature is 45-50 ℃, and the enzymolysis time is 1.5-2.5 h.

4. The method according to claim 2, wherein in step b, the yeast is a highly active dry yeast agent, and the yeast is stirred for 2-3 times per day during the light-shielding fermentation process.

5. The process according to claim 2, wherein in the step b, the fermentation is stopped when the alcoholic strength is 7 to 8 °.

6. The process according to claim 2, wherein the viable count of the acetic acid bacteria in step c is not less than 1X 10⁸cfu/g。

7. The method according to claim 2, wherein in the step d, the rotary evaporation temperature is 45-50 ℃; the freeze-drying temperature is-40 to-35 ℃.

8. Use of an elderberry extract prepared by the method of any one of claims 2 to 7 in a tabletted confection.

9. The use according to claim 8, wherein the tabletted confectionery product comprises the following components in parts by weight: 0.2-0.4 part of elderberry extract, 0.5-0.8 part of acerola cherry concentrated powder, 0.05-0.08 part of vitamin C, 7-9 parts of sorbitol, 0.1-0.3 part of citric acid, 0.005-0.007 part of aspartame, 0.2-0.3 part of magnesium stearate and 0.1-0.2 part of edible essence.

Technical Field

The invention relates to the technical field of functional food processing, in particular to a preparation method of an elderberry tablet candy and an elderberry extract.

Background

Anthocyanins, also called anthocyanidins, are water-soluble flavonoid compounds which can change color with the acid and alkali of cellular fluid and are one of the main pigments forming the color of petals and fruits. The anthocyanin is a secondary metabolite of plants, and researches show that the anthocyanin has an oxidation resistance effect which is fifty times higher than that of vitamin E and twenty times higher than that of vitamin C, is a natural antioxidant and has extremely high bioavailability. In addition, anthocyanin also has various biological activities of resisting fatigue, aging and tumor, preventing cardiovascular and cerebrovascular diseases and the like, and is widely applied to industries such as functional food, cosmetics, pharmacy and the like at present.

The elderberry is the berry of elderberry of Caprifoliaceae, is bitter, pungent and mild in flavor, is red or blue-violet black oval or nearly round, is rich in polyphenol substances and phytochemicals, can resist viruses, bacteria, nasal mucositis, diuresis, sweating promotion and the like, and the anthocyanin in the elderberry has the functions of helping to regulate immune response and enhancing human body resistance. The anthocyanin in the elderberry is mainly an anthocyanin substance taking cyanidin as a main component and an anthocyanin substance taking cyanidin as an aglycone, and is one of the plant sources containing more anthocyanin at present. Johansen isolated 4 anthocyanins from the fruit of sambucus canadensis. Nakatani et al isolated 2 novel acylated anthocyanin compounds from the fruit. At present, researches on anthocyanin at home and abroad are mostly concentrated on crops such as blueberries, purple potatoes, mulberries, black barbary wolfberry fruits and the like, and few reports are made on researches on extraction processes of the anthocyanin of the blackberries.

At present, the common method for extracting the anthocyanin is an organic solvent extraction method, the solvent is alcohol solution such as ethanol or methanol, the extraction rate can be improved by adding a small amount of hydrochloric acid or acetic acid solution, an ultrasonic auxiliary method, a microwave auxiliary method and the like, the method is simple and convenient to operate, and complex instruments and equipment are not needed; however, the use of a large amount of organic solvents during extraction results in increased production cost, easily damaged active ingredients, and poor safety due to environmental pollution caused by certain toxicity of solvents such as methanol.

Generally, mature plant cell walls are composed of cellulose, hemicellulose, pectin, and lignin. The principle of the enzymatic extraction is that hydrolytic enzymes such as cellulase, pectinase, hemicellulase and the like are used for decomposing plant cell walls, destroying cell structures and promoting the diffusion and leaching of functional components, so that the extraction rate is improved. And the enzyme extraction is carried out at a lower temperature, so that the effective components can be well protected from being damaged. The solid state fermentation refers to a fermentation process in which a reaction system is anhydrous or close to anhydrous, the treatment process of the raw materials is relatively simple, and the damage to the nutrient components and the sensory characteristics of the raw materials is small. In the fermentation process, macromolecular fibers, organic acids, pectin and other substances are fermented by microorganisms to form rich amino acids, enzymes, vitamins, mineral substances and secondary metabolites, so that the nutrition is rich and the absorption by a human body is facilitated. At present, no report about the enzymatic extraction of the elderberry anthocyanin and the preparation of the elderberry extract by using a fermentation method exists.

Disclosure of Invention

The invention aims to provide an elderberry tablet candy and a preparation method of an elderberry extract, and aims to solve the problems that the cost is high, effective components are easily damaged, and the safety is poor when the prior art is applied to extracting elderberry anthocyanin.

The technical scheme adopted by the invention is as follows: the elderberry tabletting candy comprises the following components in parts by weight: 0.2-0.4 part of elderberry extract, 0.5-0.8 part of acerola cherry concentrated powder, 0.05-0.08 part of vitamin C, 7-9 parts of sorbitol, 0.1-0.3 part of citric acid, 0.005-0.007 part of aspartame, 0.2-0.3 part of magnesium stearate and 0.1-0.2 part of edible essence. Wherein the elderberry extract is prepared by the following method.

A preparation method of elderberry extract comprises the following steps:

a. crushing elderberry into pulp, and adding cellulase and pectinase for enzymolysis;

b. inoculating 2-5wt% of yeast after enzymolysis, fermenting at 18-23 deg.C in dark place, stopping fermentation until alcoholic strength is 5.5-8.7 °, subjecting the fermentation product to ultrasonic extraction at 40-45 deg.C for 1-2 hr, and filtering to obtain primary fermentation solution;

c. pumping the primary fermentation liquid into a fermentation tank, adding 8-10 wt% of acetic acid bacteria, keeping the ventilation amount of sterile air at 0.06-0.08vvm, keeping the fermentation temperature at 28-33 ℃, stopping fermentation when the pH value is 2.8-3 after stirring and fermenting, placing the fermentation product at 40-45 ℃ for ultrasonic extraction for 1-2h, and filtering;

d. and performing rotary evaporation on the filtrate, and then freeze-drying into powder to obtain the elderberry extract.

In the step a, the dosage of the cellulase is 0.5 to 1 weight percent, the dosage of the pectinase is 0.07 to 0.1 weight percent, the enzymolysis temperature is 45 to 50 ℃, and the enzymolysis time is 1.5 to 2.5 hours.

In the step b, the yeast is a high-activity dry yeast agent, and is stirred for 2-3 times every day in the process of light-resistant fermentation.

And b, stopping fermentation until the alcoholic strength is 7-8 degrees.

In the step c, the viable count of the acetic acid bacteria is more than or equal to 1 multiplied by 10⁸cfu/g。

In the step d, the rotary evaporation temperature is 45-50 ℃; the freeze-drying temperature is-40 to-35 ℃.

The application of the elderberry extract prepared by the method in tabletting candies.

The tabletted candy comprises the following components in parts by weight: 0.2-0.4 part of elderberry extract, 0.5-0.8 part of acerola cherry concentrated powder, 0.05-0.08 part of vitamin C, 7-9 parts of sorbitol, 0.1-0.3 part of citric acid, 0.005-0.007 part of aspartame, 0.2-0.3 part of magnesium stearate and 0.1-0.2 part of edible essence.

The raw material used in the invention is elderberry, most of the documents related to the elderberry at present are preparation methods of concentrated fruit juice thereof, and no report is provided about a method for extracting anthocyanin in the elderberry.

According to the invention, after the elderberry is pulped, cellulase and pectinase are added for enzymolysis reaction, and the cell wall of the elderberry is destroyed to promote the anthocyanin to be better dissolved out. Then adding a specific amount of yeast, fermenting to generate ethanol to a specific alcoholic strength, and performing ultrasonic-assisted extraction to obtain a fermentation primary liquid. Inoculating the initial fermentation liquid into a fermentation tank, adding a specific amount of acetic acid bacteria, fermenting to produce acetic acid, stopping fermentation when the pH value is reduced to 2.8-3, and keeping the pH value within the range, wherein the anthocyanin has the best stability and can protect the effective components to the maximum extent. Finally, carrying out ultrasonic treatment again for extraction and freeze-drying into powder, wherein the content of anthocyanin in the obtained extract is up to 77.2-81.3 mg/g. The method adopts an ultrasonic-assisted solvent to extract the anthocyanin in the elderberry twice under the optimal condition, the content of the anthocyanin is only 57.3mg/g, and the extraction rate of the anthocyanin is improved by 34.7-41.9%.

When the fermentation method is adopted for extraction, no additional C source and N source are added, the nutrition of the elderberry is utilized, the fermentation cut-off index is optimized, the best effect is achieved in the shortest time, and the production period is saved. The whole preparation process does not use organic extraction solvent, has no food safety risks such as solvent residue and the like, has simple equipment, mild control conditions, simple and convenient operation compared with the traditional extraction mode, has high extraction efficiency and extraction specificity aiming at anthocyanin, is economic and environment-friendly, effectively reduces the production cost, and is beneficial to industrialized mass production. The elderberry extract prepared by the method can be widely applied to the field of food, and the invention also provides an application method of the elderberry extract in tabletting candies, and the tabletting candies have certain oxidation resistance in hydroxyl free radicals, DPPH free radicals and total reducing capacity systems.

Drawings

FIG. 1 is a graph showing the comparison of the anthocyanin content of the extracts obtained in example 1 under the action of various enzymes.

Fig. 2 is a photograph of the tabletted confectioneries prepared in examples 6 and 7.

Detailed Description

The present invention is described in detail below with reference to specific examples, wherein reagents and procedures not mentioned in the examples are all performed according to the routine procedures in the art.

In the examples, pectinase (30000U/g) and cellulase (20000U/g) were obtained from Henschel Biotechnology Ltd, yeast was obtained from Angel Yeast Ltd, and acetic acid bacteria (Shanghai brewing 1.01) were obtained from Shanghai brewing Co., Ltd.

Example 1 comparison of the Effect of different enzymes

Weighing elderberry, pulping into fruit pulp by using a homogenizer, and dividing into 4 groups, wherein the step a is that no enzyme is added; b, adding 0.5% of cellulase; c, adding 0.07 percent of pectinase; d: adding 0.5% cellulase and 0.07% pectase, and performing enzymolysis at 50 deg.C for 2 hr.

Inoculating 2% yeast according to weight of each part, fermenting at 20 deg.C in the absence of light, stirring for 2 times per day, detecting alcohol content to 7 °, performing ultrasonic extraction at 40 deg.C for 1.5 hr, and filtering to obtain primary fermentation liquid.

Pumping the primary fermentation liquid into a fermentation tank, inoculating acetobacter with the total weight of 8%, fermenting at the conditions of ventilation volume of 0.08vvm and 30 ℃, monitoring the pH value, reducing to 3, performing ultrasonic extraction at 40 ℃ for 1.5h, and filtering to obtain fermentation liquid.

Performing rotary evaporation and concentration on the fermentation liquor at 45 ℃, freeze-drying at-35 ℃ for 24h, crushing and sieving to obtain the elderberry extract. The results of the detection of the obtained extract are shown in figure 1, and the addition of cellulase and pectinase can effectively promote the dissolution of anthocyanin and improve the extraction rate, but the effect is not as good as that of the compound enzyme of the two. The single cellulase is used for enzymolysis, and the anthocyanin content is improved by 12.9 percent; the anthocyanin content is improved by 10.2 percent by single pectinase for enzymolysis; the anthocyanin content is improved by 14.8 percent through the enzymolysis of the compound enzyme. Therefore, the enzymolysis is carried out by adopting the complex enzyme of the cellulase and the pectinase.

Example 2

Weighing 50kg of elderberry, pulping into fruit pulp by using a homogenizer, placing in parts, adding 0.5% of cellulase and 0.07% of pectinase according to the weight of each part respectively, and performing enzymolysis for 2h at 50 ℃.

Adding yeast 1% by weight, fermenting at 18 deg.C in the absence of light, stirring for 2 times per day, detecting alcohol content to 6 °, performing ultrasonic extraction at 40 deg.C for 1.5 hr, and filtering to obtain primary fermentation liquid.

Pumping the primary fermentation liquid into a fermentation tank, inoculating acetobacter with the total weight of 8%, fermenting at the conditions of ventilation volume of 0.06vvm and 28 ℃, monitoring the pH value, reducing to 2.8, performing ultrasonic extraction at 40 ℃ for 1.5h, and filtering to obtain fermentation liquid.

Performing rotary evaporation and concentration on the fermentation liquor at 50 ℃, freeze-drying for 24h at-35 ℃, and crushing and sieving to obtain the elderberry extract.

Through detection, the content of anthocyanin in the elderberry extract is 77.2 mg/g. Compared with the anthocyanin content in the extract obtained by the ultrasonic-assisted solvent extraction method of 57.3mg/g, the anthocyanin content is improved by 34.7%.

Example 3

Weighing 100kg of elderberry, pulping into fruit pulp by using a homogenizer, placing in parts, adding 0.7% of cellulase and 0.1% of pectinase according to the weight of each part respectively, and performing enzymolysis for 2h at 45 ℃.

Inoculating 2% of yeast according to the weight of each part, fermenting under the conditions of no light and 20 ℃, stirring for 2 times per day on time, detecting that the alcoholic strength reaches 8.7 degrees, performing ultrasonic extraction for 1.5h at 45 ℃, and filtering to obtain a primary fermentation solution.

Pumping the primary fermentation liquid into a fermentation tank, inoculating acetobacter with the total weight of 10%, fermenting at the conditions of ventilation volume of 0.06vvm and 30 ℃, monitoring the pH value, reducing to 2.9, performing ultrasonic extraction at 45 ℃ for 1.5h, and filtering to obtain fermentation liquid.

Performing rotary evaporation and concentration on the fermentation liquor at 45 ℃, freeze-drying at-40 ℃ for 24h, crushing and sieving to obtain the elderberry extract.

Through detection, the content of anthocyanin in the elderberry extract is 79.4 mg/g. Compared with the anthocyanin content in the extract obtained by the ultrasonic-assisted solvent extraction method of 57.3mg/g, the anthocyanin content is improved by 38.6 percent.

Example 4

Weighing 200kg of elderberry, pulping into fruit pulp by using a homogenizer, placing in parts, adding 1.0% of cellulase and 0.1% of pectinase according to the weight of each part respectively, and performing enzymolysis for 2h at 45 ℃.

Inoculating 5% of yeast according to weight of each part, fermenting at 23 deg.C in the absence of light, stirring for 3 times per day, detecting alcohol content to 7.5 °, performing ultrasonic extraction at 45 deg.C for 1.5 hr, and filtering to obtain primary fermentation liquid.

Pumping the primary fermentation liquid into a fermentation tank, inoculating acetobacter with the total weight of 10%, fermenting at the conditions of ventilation volume of 0.08vvm and 33 ℃, monitoring the pH value, reducing to 3, performing ultrasonic extraction at 45 ℃ for 1.5h, and filtering to obtain fermentation liquid.

Performing rotary evaporation and concentration on the fermentation liquor at 45 ℃, freeze-drying at-40 ℃ for 24h, crushing and sieving to obtain the elderberry extract.

Through detection, the content of anthocyanin in the elderberry extract is 82.3 mg/g. Compared with the anthocyanin content in the extract obtained by the ultrasonic-assisted solvent extraction method of 57.3mg/g, the anthocyanin content is improved by 41.9%.

Example 5

Weighing 50kg of elderberry, pulping into fruit pulp by using a homogenizer, placing in parts, adding 0.5% of cellulase and 0.07% of pectinase according to the weight of each part respectively, and performing enzymolysis for 2h at 50 ℃.

Adding yeast 1% by weight, fermenting at 18 deg.C in the absence of light, stirring for 2 times per day, detecting alcohol content to 4 °, performing ultrasonic extraction at 40 deg.C for 1.5 hr, and filtering to obtain primary fermentation liquid.

Pumping the primary fermentation liquid into a fermentation tank, inoculating acetobacter with the total weight of 8%, fermenting at the conditions of ventilation volume of 0.06vvm and 28 ℃, monitoring the pH value to 3.5, performing ultrasonic extraction at 40 ℃ for 1.5h, and filtering to obtain fermentation liquid.

Performing rotary evaporation and concentration on the fermentation liquor at 50 ℃, freeze-drying for 24h at-35 ℃, and crushing and sieving to obtain the elderberry extract.

Through detection, the content of anthocyanin in the elderberry extract is 71.3 mg/g. Compared with the example 1, the fermentation cut-off index is changed, other conditions are not changed, and the extraction rate is reduced by 7.6%, so that the strict control of the fermentation cut-off index is very important.

Example 6

According to the weight portion, 0.2 portion of elderberry extract, 0.5 portion of acerola cherry concentrated powder, 0.05 portion of vitamin C, 8.845 portions of sorbitol, 0.1 portion of citric acid, 0.005 portion of aspartame, 0.2 portion of magnesium stearate and 0.1 portion of edible essence are weighed.

After the raw materials are qualified, the obtained tablets are complete and smooth, have uniform color and no visible foreign matters with normal vision through the processes of weighing, mixing and tabletting, and meet various regulations, as shown in figure 2.

Example 7

According to the weight portion, 0.4 portion of elderberry extract, 0.8 portion of acerola cherry concentrated powder, 0.08 portion of vitamin C, 7.913 portions of sorbitol, 0.3 portion of citric acid, 0.007 portion of aspartame, 0.3 portion of magnesium stearate and 0.2 portion of edible essence are weighed.

After the raw materials are qualified, the obtained tablets are complete and smooth, have uniform color and no visible foreign matters with normal vision through the processes of weighing, mixing and tabletting, and meet various regulations, as shown in figure 2.

Example 8 in vitro antioxidant Effect of tabletted confections

The tabletted candies of example 6 and example 7 were each taken and tabletted several times, crushed and dissolved in a suitable amount of purified water, and filtered to obtain a sample solution.

Adding 1mL of the sample solution into a test tube respectively, and adding 8mmol/L FeSO₄And 8mmol/L salicylic acid-ethanol solution each 1mL, mixing well, adding 1mLH₂O₂Reacting the solution at 37 deg.C for 30min, measuring absorbance at 520nm, and recording as A₁(ii) a Using distilled water as reference control instead of salicylic acid solution, and measuring absorbance value A₂(ii) a Using distilled water as a blank control instead of the sample solution, and measuring the absorbance as A₀. The test was repeated three times, and the hydroxyl radical clearance of the two sample solutions was determined to be 59.32% and 77.24%, respectively.

Adding 2mL of the sample solution into a test tube, adding 2mL of 6mmol/L DPPH ethanol solution, mixing, standing in a dark room for 30min, and measuring absorbance at 517nmValue, denoted as A₁(ii) a Replacing sample solution with purified water, and measuring absorbance value A₀. The three experiments were repeated and the DPPH radical clearance was found to be 71.92% and 89.45% for the two sample solutions, respectively.

Adding 1mL of the sample solution into a test tube, sequentially adding 1mL of each of 0.2mol/L phosphate buffer solution (pH =6.6) and 1% potassium ferricyanide solution, uniformly mixing, reacting in a water bath at 50 ℃ for 20min, rapidly cooling, adding 1mL of 10% trichloroacetic acid solution, and centrifuging for 10min at 5000 r/min. Taking 2mL of the supernatant fluid, adding 2mL of distilled water and 0.4mL of 0.1% ferric trichloride solution into a test tube, fully and uniformly mixing, standing in a dark room for reaction for 10min, taking the distilled water as a blank for zero adjustment, and measuring the absorbance value at 700 nm. The test was repeated three times, and the absorbance values of the two sample solutions were measured to be 1.84 and 2.76, respectively.

The tabletted confectionery products exhibit a certain antioxidant capacity in hydroxyl radicals, DPPH radicals and total reducing power systems.