Preparation method and application of cocoa shell color

Abstract

The invention provides a preparation method and application of cocoa shell color. The preparation method comprises the following steps: crushing cocoa shells, immersing the crushed cocoa shells into a container filled with mixed enzymatic hydrolysate, carrying out enzymolysis in a constant-temperature dark place, filtering, adding alkali liquor, regulating the pH value, carrying out centrifugal separation, heating and concentrating, regulating the pH value, filtering and sterilizing, and drying to obtain a cocoa shell color finished product. The mixed enzymatic hydrolysate is an aqueous solution of cellulase RS, hemicellulase and macerozyme R10. The production efficiency of the cocoa shell color is high, the operation process is simple and convenient, the production cost is low, pollution is avoided, the black brown or dark brown high-purity cocoa shell color powder with the color value larger than 50 can be stably obtained, the stability of the cocoa shell color is good, and the cocoa shell color powder can be used for adding food pigments.

Description

Technical Field

[0001] The invention relates to the field of preparation of cocoa husk color, in particular to a preparation method of cocoa husk color and application thereof. Background Art

[0002] Cocoa peel accounts for about 28-30% of the dry weight of cocoa fruit, contains a variety of active substances such as flavonoids, anthocyanins, chlorogenic acid, etc., and has antioxidant and anti-inflammatory effects. However, only a small amount of cocoa peel is reused, and most of it is discarded as waste, which not only causes a waste of resources, but also has adverse effects on the environment. Cocoa shell color is a brown pigment formed by oxidation or condensation of substances such as catechins, anthocyanins, and anthocyanins contained in cocoa beans and their outer shells during fermentation and baking. Cocoa shell color is a natural edible pigment, which is a brown powder, odorless, slightly bitter, easy to absorb moisture, and easily soluble in water and dilute ethanol solutions. Therefore, improving the reuse value of cocoa peel and realizing the efficient utilization of cocoa peel can promote the healthy development of the cocoa industry. At present, the extraction and application of cocoa shell color have the following problems: first, the extraction rate is low and there are residues. Since the leaching solution uses a high-concentration sodium hydroxide solution, it may cause adverse environmental impacts. Second, there are many operating steps and the process is complicated. Currently, multi-stage countercurrent extraction and gradient elution are used, which increases expenses and costs, and the adsorption rate is extremely low, resulting in low product yield and purity. Summary of the invention

[0003] In view of this, the present invention provides a preparation method of cocoa shell color and application thereof to solve the above problems.

[0004] The technical solution of the present invention is achieved in this way: A method for preparing cocoa shell color comprises the following steps: S1, taking cocoa shells, crushing them and immersing them in a container containing a mixed enzymatic hydrolysate, wherein the mixed enzymatic hydrolysate consists of cellulase RS, hemicellulase, mesenzyme R10 and water, performing enzymatic hydrolysis at a constant temperature and avoiding light, filtering and taking the supernatant to obtain cocoa shell powder enzymatic hydrolysate; S2, adding alkali solution to the cocoa shell powder enzymatic hydrolyzate, soaking and filtering, adjusting the pH value, centrifuging, taking the supernatant, and obtaining a centrifugal supernatant; S3, heating and concentrating the centrifugal supernatant, standing, and filtering to obtain a pigment solution; S4, concentrating the pigment solution, adjusting the pH value, filtering, sterilizing, and drying to obtain the cocoa shell color finished product.

[0005] Furthermore, in step S1, the cocoa shell is crushed and then passed through a 80-100 mesh sieve.

[0006] Furthermore, in step S1, the mixed enzymatic solution is cellulase RS, hemicellulase, macerate R10 and water in a mass volume ratio of 1-1.5 g:2-3 g:1-1.5 g:80-100 mL.

[0007] Furthermore, in step S1, the rotation speed of the constant temperature stirring is 65-70 rpm, the enzymolysis temperature is 35-40° C., the enzymolysis time is 3-4 h, and the enzymolysis pH value is 4.8-5.5.

[0008] Further, in step S2, the alkali solution is composed of an aqueous sodium hydroxide solution and an aqueous sodium carbonate solution in a volume ratio of 1-3:1, the mass percentage of the aqueous sodium hydroxide solution is 15%-25%, the mass percentage of the aqueous sodium carbonate solution is 10%-20%, and the volume ratio of the enzymatic hydrolyzate to the alkali solution is 1:0.2-0.4.

[0009] Furthermore, in step S2, the pH value is adjusted to 9-11, and in step S4, the pH value is adjusted to 6-7.5.

[0010] Furthermore, in step S3, the concentration is nanofiltration concentration, and the molecular weight of the nanofiltration membrane used is 200-2000.

[0011] Furthermore, in step S4, the sterilization temperature is 85-95° C. and the sterilization time is 40-50 min.

[0012] Compared with the prior art, the present invention has the following beneficial effects: The invention crushes the cocoa shell and then performs enzymolysis, adjusts the pH value twice, and concentrates and dries to obtain a cocoa shell color product. The invention has a simple and convenient operation process for producing cocoa shell color, low production cost, high production efficiency, and no pollution. The enzymolysis step increases the color value of the cocoa shell color, and the control of the pH value increases the acid dissolution maintenance rate and the salt dissolution maintenance rate of the cocoa shell color, thereby increasing the stability of the cocoa shell color. DETAILED DESCRIPTION

[0013] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0014] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods, and the reagents used in the examples are all available from commercial sources.

[0015] Cellulase RS ( Cellulase RS ), freeze-dried powder, cellulase activity>16000U / g, number 61003ES08, manufacturer is Yisheng Biotechnology (Shanghai) Co., Ltd.; hemicellulase ( Hemicellulase), CAS No. 9025-56-3, BR activity ≥ 5unit / mg, manufacturer is Shanghai Yanjin Biotechnology Co., Ltd.; Isolase R10 ( Macerozyme R-10 ), CAS number is 9032-75-1, enzyme activity ≥3000u / g, manufacturer is Shanghai ELISA Biotechnology Co., Ltd.

[0016] Example 1 A method for preparing cocoa shell color comprises the following steps: Step (1): Take clean cocoa shells and crush them, then pass them through an 80-mesh sieve. The sieved cocoa shell powder is immersed in a container filled with a mixed enzymatic solution, wherein the mixed enzymatic solution is cellulase RS, hemicellulase, cleavage enzyme R10 and water in a mass volume ratio of 1g:2g:1g:80mL. The cocoa shell powder is enzymatically hydrolyzed in a constant temperature and light-proof stirring environment and then filtered. During the enzymatic hydrolysis, the constant temperature stirring speed is 65rpm, the enzymatic hydrolysis temperature is 35°C, the enzymatic hydrolysis time is 3h, and the enzymatic hydrolysis pH is 4.8. After the enzymatic hydrolysis, the supernatant is taken to obtain the cocoa shell powder enzymatic hydrolyzate; Step (2): adding cocoa shell powder enzymatic hydrolysate to an alkali solution, wherein the alkali solution is composed of a sodium hydroxide aqueous solution and a sodium carbonate aqueous solution in a volume ratio of 1:1, the mass percentage of the sodium hydroxide aqueous solution is 15%, the mass percentage of the sodium carbonate aqueous solution is 10%, and the volume ratio of the enzymatic hydrolysate to the alkali solution is 1:0.2; after adding the alkali solution to the cocoa shell powder enzymatic hydrolysate, the cocoa shell powder enzymatic hydrolysate is soaked and filtered, the pH value is adjusted to 9, centrifuged, and the supernatant is taken to obtain a cocoa shell powder centrifugal supernatant; Step (3): heating the supernatant of the cocoa shell powder centrifuged and concentrating it by nanofiltration, the molecular weight of the nanofiltration membrane is 800, and filtering it after concentration to obtain a pigment solution; Step (4): Concentrate the pigment solution, adjust the pH value to 6, filter, sterilize at a temperature of 85° C. for 40 min, and dry after sterilization to obtain a cocoa shell color product.

[0017] Example 2 A method for preparing cocoa shell color comprises the following steps: Step (1): Take clean cocoa shells and crush them, then pass them through an 80-mesh sieve. The sieved cocoa shell powder is immersed in a container filled with a mixed enzymatic hydrolysis solution, wherein the mixed enzymatic hydrolysis solution is cellulase RS, hemicellulase, mesenzyme R10 and water in a mass volume ratio of 1g:3g:1.5g:80mL. The cocoa shell powder is enzymatically hydrolyzed in a constant temperature and light-proof stirring environment and then filtered. During the enzymatic hydrolysis, the constant temperature stirring speed is 70rpm, the enzymatic hydrolysis temperature is 40°C, the enzymatic hydrolysis time is 4h, and the enzymatic hydrolysis pH is 5. After the enzymatic hydrolysis, the supernatant is taken to obtain the cocoa shell powder enzymatic hydrolysis solution; Step (2): adding cocoa shell powder enzymatic hydrolysate to an alkali solution, wherein the alkali solution is composed of a sodium hydroxide aqueous solution and a sodium carbonate aqueous solution in a volume ratio of 3:1, the mass percentage of the sodium hydroxide aqueous solution is 25%, the mass percentage of the sodium carbonate aqueous solution is 12%, and the volume ratio of the enzymatic hydrolysate to the alkali solution is 1:0.2; after adding the alkali solution to the cocoa shell powder enzymatic hydrolysate, the cocoa shell powder enzymatic hydrolysate is soaked and filtered, the pH value is adjusted to 11, centrifuged, and the supernatant is taken to obtain a cocoa shell powder centrifugal supernatant; Step (3): heating the supernatant of the cocoa shell powder centrifuged and concentrating it by nanofiltration, the molecular weight of the nanofiltration membrane is 1000, and filtering it after concentration to obtain a pigment solution; Step (4): Concentrate the pigment solution, adjust the pH value to 7.5, filter, sterilize at a temperature of 95° C. for 50 min, and dry after sterilization to obtain a cocoa shell color product.

[0018] Example 3 A method for preparing cocoa shell color comprises the following steps: Step (1): Take clean cocoa shells and crush them, then pass them through an 80-mesh sieve. The sieved cocoa shell powder is immersed in a container filled with a mixed enzymatic hydrolysis solution, wherein the mixed enzymatic hydrolysis solution is cellulase RS, hemicellulase, cleavage enzyme R10 and water in a mass volume ratio of 1g:2g:1.5g:100mL. The cocoa shell powder is enzymatically hydrolyzed in a constant temperature and light-proof stirring environment and then filtered. During the enzymatic hydrolysis, the constant temperature stirring speed is 70rpm, the enzymatic hydrolysis temperature is 35°C, the enzymatic hydrolysis time is 3.5h, and the enzymatic hydrolysis pH is 5.0. After the enzymatic hydrolysis, the supernatant is taken to obtain the cocoa shell powder enzymatic hydrolysis solution; Step (2): adding cocoa shell powder enzymatic hydrolysate to an alkali solution, wherein the alkali solution is composed of a sodium hydroxide aqueous solution and a sodium carbonate aqueous solution in a volume ratio of 2:1, the mass percentage of the sodium hydroxide aqueous solution is 20%, the mass percentage of the sodium carbonate aqueous solution is 15%, and the volume ratio of the enzymatic hydrolysate to the alkali solution is 1:0.3; after adding the alkali solution to the cocoa shell powder enzymatic hydrolysate, the cocoa shell powder enzymatic hydrolysate is soaked and filtered, the pH value is adjusted to 10, centrifuged, and the supernatant is taken to obtain a cocoa shell powder centrifugal supernatant; Step (3): heating the supernatant of the cocoa shell powder centrifuged and concentrating it by nanofiltration, the molecular weight of the nanofiltration membrane is 1000, and filtering it after concentration to obtain a pigment solution; Step (4): Concentrate the pigment solution, adjust the pH value to 7, filter, sterilize at a temperature of 90° C. for 45 min, and dry after sterilization to obtain a cocoa shell color product.

[0019] Comparative Example 1 The main difference between Comparative Example 1 and Example 3 is that the enzymatic hydrolysis step is removed from step (1) of Example 3, and the preparation methods of other steps (2) to (4) remain unchanged.

[0020] Comparative Example 2 The main difference between Comparative Example 2 and Example 3 is that the mass volume ratio of the enzymatic hydrolysate in step (1) in Example 3 is changed to 1 g:0.2 g:0.2 g:100 mL of cellulase RS, hemicellulase, cleavage enzyme R10 and water, and the nanofiltration concentration in step (3) is changed to heating evaporation concentration, and the preparation methods of other steps (1) to (4) remain unchanged.

[0021] Comparative Example 3 The main difference between Comparative Example 3 and Example 3 is that the step of adjusting the pH value is removed from step (2) and step (4) of Example 3, and the preparation method of other steps (1) to step (4) remains unchanged.

[0022] Comparison Commercially available cocoa shell color was used, and the manufacturer was Henan Kangzhiwang Biotechnology Co., Ltd.

[0023] test According to the method A.6 in Appendix A of GB1886.30 "Food Safety Standard Food Additive Cocoa Shell Color", the color value of the natural cocoa shell color powder and the control product prepared by the methods of Examples 1-3 and Comparative Examples 1-3 was measured and calculated. The specific operation method is: weigh 0.1000g of cocoa shell color, dissolve it with distilled water and dilute it to 100 mL, shake it well; use a pipette to draw 10 ml of the sample solution in the shaking state, and then dilute it to 100 mL (that is, 0.1% aqueous solution), use a spectrophotometer, use water as a reference solution, measure its absorbance at a wavelength of 400 nm in a 1 cm cuvette, and calculate the color value.

[0024] The formula for calculating the color price of the finished product is E 1%(400nm) =A×f / (100×m), A is the absorbance of the test sample; f is the dilution factor; m is the mass of the sample.

[0025] Table 1 Acid dissolution maintenance test: Under the condition of maintaining the temperature at 15°C and 25°C, 10 ml of 0.3% citric acid aqueous solution was added to the cocoa pigments of Examples 1-3, Comparative Examples 1-3 and the control, and stirred until the color value was 0.1 (E 10 ％(400nm) ). The mixture was allowed to stand for 24 hours, centrifuged at 3000 rpm for 30 minutes, and the cocoa pigment was diluted with a citric acid buffer solution with a pH value of 7. The concentration of cocoa pigment was measured by a spectrophotometer.

[0026] Acid dissolution maintenance rate (%) = dissolved cocoa pigment / total amount of cocoa pigment*100%.

[0027] Salt dissolution maintenance test: Under the condition of maintaining the temperature at 15°C and 25°C, 10 ml of 10% sodium chloride aqueous solution was added with cocoa pigments of Examples 1-3, Comparative Examples 1-3 and the control and stirred to make the color value 0.1 (E 10 ％(400nm) ). The mixture was allowed to stand for 24 hours, centrifuged at 3000 rpm for 30 minutes, and the cocoa pigment was diluted with a citric acid buffer solution with a pH value of 7. The concentration of cocoa pigment was measured by a spectrophotometer.

[0028] Salt dissolution maintenance rate (%) = dissolved cocoa pigment / total amount of cocoa pigment*100%.

[0029] Table 2 Group 15℃ acid dissolution maintenance rate (%) 15℃ Salt dissolution maintenance rate (%) 25℃ acid dissolution maintenance rate (%) 25℃ Salt dissolution maintenance rate (%) Example 1 87.2 95.6 87.9 96.5 Example 2 86.9 94.1 87.7 94.9 Example 3 87.5 96.6 88.5 97.4 Comparative Example 1 73.0 82.3 74.1 82.8 Comparative Example 2 76.8 78.4 77.2 79.3 Comparative Example 3 72.4 80.5 73.9 81.7 Comparison 72.0 82.2 72.8 83.1 The test results show that the cocoa shell color prepared by the method of the present invention can obtain a dark brown or dark brown cocoa shell color powder with a finished cocoa shell color color value of 54.6-56.7, and the color value is greater than 50. After the enzymatic hydrolysis step is removed in the preparation method of comparative example 1, the color value of the finished product is reduced to 32.4; after the mass volume ratio of the enzymatic hydrolyzate is modified and the concentration process is modified in the preparation method of comparative example 2, the color value of the finished cocoa shell color is reduced to 33.5; the pH adjustment step is removed in the preparation method of comparative example 2, which also reduces the color value, indicating that the preparation method of the cocoa shell color of the present invention is beneficial to improving the color value of the finished cocoa shell color. At the same time, the analysis of the acid solubility maintenance rate and the salt solubility maintenance rate of the cocoa shell color shows that the cocoa shell color prepared by the method of the present invention is relatively stable, relatively acid-resistant and salt-resistant, and not easy to deteriorate. In summary, the present invention has high production efficiency for producing cocoa shell color, simple process, convenient operation, low production cost, no pollution, can stably obtain black brown or dark brown high-purity cocoa shell color powder with a color value greater than 50, and the cocoa shell color has good stability, and can be used for adding food pigments.

[0030] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing cocoa shell color, characterized in that: The following steps are involved: S1, taking cocoa shells, crushing them and immersing them in a container containing a mixed enzymatic hydrolysate, wherein the mixed enzymatic hydrolysate consists of cellulase RS, hemicellulase, mesenzyme R10 and water, performing enzymatic hydrolysis at a constant temperature and avoiding light, filtering and taking the supernatant to obtain cocoa shell powder enzymatic hydrolysate; S2, adding the cocoa shell powder enzymatic hydrolyzate to the alkali solution, soaking and filtering, adjusting the pH value, centrifuging, taking the supernatant, and obtaining the centrifuged supernatant; S3, heating and concentrating the centrifugal supernatant, standing, and filtering to obtain a pigment solution; S4, concentrating the pigment solution, adjusting the pH value, filtering, sterilizing, and drying to obtain the cocoa shell color finished product.

2. The method for preparing cocoa shell color according to claim 1, characterized in that: Step S1, the cocoa shell is crushed and passed through a 80-100 mesh sieve after being crushed.

3. The method for preparing cocoa shell color according to claim 1, characterized in that: Step S1, the mass volume ratio of the cellulase RS, hemicellulase, macerate R10 and water is 1-1.5g:2-3g:1-1.5g:80-100mL of cellulase RS, hemicellulase, macerate R10 and water.

4. The method for preparing cocoa shell color according to claim 1, characterized in that: Step S1, the rotation speed of the constant temperature stirring is 65-70 rpm, the enzymolysis temperature is 35-40°C, the enzymolysis time is 3-4h, and the enzymolysis pH value is 4.8-5.

5.

5. The method for preparing cocoa shell color according to claim 1, characterized in that: Step S2, the alkali solution is composed of a sodium hydroxide aqueous solution and a sodium carbonate aqueous solution in a volume ratio of 1-3:1, the mass percentage of the sodium hydroxide aqueous solution is 15%-25%, the mass percentage of the sodium carbonate aqueous solution is 10%-20%, and the volume ratio of the enzymatic hydrolyzate to the alkali solution is 1:0.2-0.

4.

6. The method for preparing cocoa shell color according to claim 1, characterized in that: In step S2, the pH value is adjusted to 9-11, and in step S4, the pH value is adjusted to 6-7.

5.

7. The method for preparing cocoa shell color according to claim 1, characterized in that: In step S3, the concentration is nanofiltration concentration, and the molecular weight of the nanofiltration membrane used is 200-2000.

8. The method for preparing cocoa shell color according to claim 1, characterized in that: Step S4, the sterilization temperature is 85-95°C and the sterilization time is 40-50min.

9. Use of the cocoa shell color prepared according to claims 1-8 in the coloring of foods, medicines, health products and fine chemical products.

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