CN116730827A - Process for extracting carnosic acid and application of carnosic acid extracted by process in vegetable oil - Google Patents

Abstract

This application relates to a process for extracting carnosic acid and the application of the extracted carnosic acid in vegetable oil. S1. Add rosemary leaves to a mixed solution of ethanol and water at the first concentration for heating and extraction. The extraction is completed. After that, the first-stage extraction liquid is obtained; S2, the first-stage extraction liquid is concentrated, the concentrated liquid is filtered by plate and frame, and the filter cake is washed with water to obtain the filtrate and filter residue; S3, the filtrate is extracted, and the extract is concentrated. , after drying, the crude rosmarinic acid product can be obtained; S4. Re-extract the filter residue with a mixed solution of ethanol and water with a second step concentration. After the re-extraction is completed, filter to obtain the filtrate and filter residue; the filter residue is Uva Ursi. Crude extract of carnosic acid and carnosic acid; S5, decolorize the filtrate, and then concentrate it into a paste. After vacuum drying, the crude product of carnosic acid is obtained. The method in this application can separate a variety of active substances and obtain a variety of crude products, which can improve the utilization rate of rosemary and thereby reduce production costs.

Description

A process for extracting carnosic acid and the use of the extracted carnosic acid in vegetable oil application

Technical field

The present application relates to the technical field of drug extraction and application, and in particular to a process for extracting carnosic acid and the application of the extracted carnosic acid in vegetable oil.

Background technique

As people pay more and more attention to food safety, they begin to realize the potential harm of chemically synthesized antioxidants to humans. Advocating nature, returning to nature, and pursuing safe, non-toxic green functional antioxidants have become the mainstream in the world. With the advancement of science and technology, natural antioxidants have flourished, and people are beginning to favor plant-derived antioxidants. Compared with chemically synthesized antioxidants, some natural antioxidants show stronger antioxidant effects, and thus have attracted the attention of the international academic community. Therefore, natural antioxidants have become one of the most active fields in the antioxidant-related industry. Most of the natural antioxidants are polyphenols, among which dozens of widely used ones include rosemary extract, vitamin E, bamboo leaf flavonoids, tea polyphenols, ginkgo extract, licorice extract, etc. The active ingredients with antioxidant effects in rosemary are carnosic acid (CA), rosmarinol, rosmarinic acid (RA), carnosol, epirosmanol, etc. Among the diterpene phenolic compounds, carnosic acid has the strongest antioxidant capacity.

Carnosic acid, a phenolic diterpenoid fat-soluble compound, has a molecular formula of C ₂₀ H ₂₈ O ₄ and a relative molecular mass of 332.43. Carnosic acid appears as a yellow-green powder and is easily soluble in chloroform and acetone. Due to its safety, high efficiency, heat resistance and other properties, it is widely used in food additives, daily chemicals, feed, pharmaceutical and health products and other fields.

There are many ways to extract carnosic acid from rosemary. Some of the existing improvement methods are mainly focused on improving the purity of carnosic acid. Therefore, in the actual extraction method, other active extracts (such as amylose) are always sacrificed. Marianic acid and ursolic acid, etc.), but in practical applications, crude carnosic acid has good anti-inflammatory, antioxidant and other effects, and other active substances also have other special effects. The price of raw material rosemary It is not cheap either, so it is necessary to provide a method that can achieve the extraction of multiple active substances.

Contents of the invention

In order to improve the utilization rate of rosemary, this application provides a process for extracting carnosic acid and the application of the extracted carnosic acid in vegetable oil.

The first aspect, a process for extracting carnosic acid, adopts the following technical solution:

A process for extracting carnosic acid includes the following steps:

S1. Add rosemary leaves to a mixed solution of ethanol and water with a first-level concentration for heating and extraction. After the extraction is completed, the first-level extraction solution is obtained;

S2. Concentrate the first-stage extraction liquid in step S1, perform plate and frame press filtration on the concentrated liquid, and wash the filter cake with water to obtain the filtrate and filter residue;

S3. Extract the filtrate in step S2, concentrate the extract, and dry it to obtain crude rosmarinic acid;

S4. Re-extract the filter residue in step S2 with a mixed solution of ethanol and water of the second step concentration. After the re-extraction is completed, filter it to obtain the filtrate and filter residue; the filter residue is the crude extraction of ursolic acid and carnosol. thing;

S5: Decolorize the filtrate in step S4, and then concentrate it into a paste. After vacuum drying, the crude carnosic acid product can be obtained.

By adopting the above technical solution, in this application, the first stage of extraction with ethanol (high concentration) and water can be used to release a large amount of rosemary active substances, and then the extract is concentrated. During the concentration process, ethanol will The evaporation rate of water is faster, and the evaporation rate of water is slower. Rosmarinic acid is soluble in water, while carnosic acid, carnosol and ursolic acid are insoluble in water. Therefore, as the evaporation proceeds, the ethanol content Significantly reduce (to basically no ethanol), carnosic acid, carnosol and ursolic acid will precipitate during the concentration process, while rosmarinic acid will remain in the liquid, so during plate pressure filtration, the filtrate The main content is rosmarinic acid, and the filter residue is a mixture of carnosic acid, carnosol and ursolic acid; then a mixed solution of ethanol (lower concentration) and water with a second step concentration is used, mainly using The solubilities of carnosic acid, carnosol and ursolic acid are different in lower concentration ethanol. The solubility of carnosic acid is the largest, while the solubility of carnosolic acid and ursolic acid is small. When re-extraction, , the filtrate is mainly carnosic acid, and the filter residue is mainly carnosol and ursolic acid, so carnosic acid can be separated. The extraction method in this application is relatively simple and can extract multiple active substances of rosemary at the same time, thereby improving the utilization rate of rosemary.

Preferably, in step S1, the mass concentration of ethanol in the mixed solution of ethanol and water at the first concentration is 85-95%; the number of extractions is 2-3 times, the extraction time is 0.5-1.5h, and the extraction temperature is 65~75℃, during each extraction, add a mixed solution 8~12 times the mass of rosemary.

By adopting the above technical solution, the concentration of ethanol in the mixed solution used for extraction in this application must be greater than 85%. If the concentration is too low, carnosol and ursolic acid will be difficult to dissolve, which will lead to incomplete extraction; the number of extractions and The amount of extraction solution added can be slightly adjusted according to the actual situation, so as to ensure the extraction effect without wasting too much extraction solution.

Preferably, in the step S2, vacuum concentration is used for concentration, and the first-stage extraction liquid is concentrated to 5-15% of the original volume; ethanol is recovered during the concentration process; the filtrate includes filtered liquid and liquid collected by washing.

By adopting the above technical solution, in this application, concentration is carried out through vacuum and reduced pressure. It is preferable to evaporate a large amount of ethanol, and the remaining liquid in the concentrated solution is basically water. Therefore, carnosic acid, carnosol and ursolic acid will be evaporated. A large amount of precipitation forms a filter residue, and rosmarinic acid is water-soluble, so it can be separated from other active components.

Preferably, in step S3, the extraction agent is ethyl acetate.

By adopting the above technical solution, in this application, by further extracting with ethyl acetate, impurities can be further removed, the purity of rosmarinic acid can be improved, and a higher purity crude rosmarinic acid can be obtained.

Preferably, in step S4, the mass concentration of ethanol in the mixed solution of ethanol and water of the second step concentration is 55-65%; the amount of mixed solvent added is 25-30 times the mass of the filter residue, and the re-extraction temperature is room temperature , the recovery time is 0.5~1.5h.

By adopting the above technical solution, in this application, by controlling the concentration and temperature of ethanol in the extraction solution during re-extraction, the dissolution amount of carnosol and ursolic acid can be greatly reduced, so that the solution mainly contains carnosic acid, thereby Realize the separation of carnosic acid and carnosol.

Preferably, in step S5, activated carbon is used as the decolorizing agent.

By adopting the above technical solution, in this application, by using activated carbon for decolorization, colored impurities in the filtrate can be removed and the product phase of carnosic acid can be improved.

In the second aspect, crude carnosic acid is extracted through the above extraction method.

The third aspect is the application of the crude carnosic acid in improving the antioxidant effect in vegetable oils.

Preferably, before the crude carnosic acid is added to vegetable oil, it needs to be pre-treated, which specifically includes the following steps:

Dissolve crude carnosic acid in 90 to 95% ethanol, then add vegetable oil 3 to 4 times the mass of carnosic acid, stir and mix, vacuum concentrate and remove ethanol and water under reduced pressure, cool, and filter to remove precipitates. material, and finally add new vegetable oil to it to adjust the mass concentration of carnosic acid to 10% to obtain carnosic acid oil. Carnosic acid oil can be added to vegetable oil as an antioxidant additive to play an antioxidant role.

By adopting the above technical solution, the vegetable oil containing carnosic acid in this application is first completely dissolved in high-concentration ethanol to completely dissolve the crude product, and then mixed with a small amount of vegetable oil, and then after removing the ethanol, it is further filtered to remove non-oil-soluble impurities, so that This method can further improve the ability of carnosic acid to reduce impurities and ensure the antioxidant properties of the oil. It is also mixed with vegetable oil first to avoid directly adding it to the vegetable oil to precipitate impurities and affect the quality of the oil.

To sum up, this application includes at least one of the following beneficial technical effects:

1. In this application, by taking advantage of the solubility differences among the various active substances in rosemary, various active substances can be separated through operations such as extraction, concentration, and filter residue re-extraction, and a variety of crude products can be obtained correspondingly. This will not be due to Extracting carnosic acid and affecting the extraction of other active substances can greatly improve the utilization rate of rosemary, thereby reducing production costs.

2. The crude carnosic acid in this application can be directly applied to vegetable oils to play an antioxidant role, which can significantly improve the antioxidant effect of vegetable oils. There is no need for further purification. It is only necessary to combine the crude carnosic acid with vegetable oil before use. After blending, it can be added to vegetable oil as an additional component.

Description of drawings

Figure 1: Peroxide value change curve of carnosic acid soybean oil in application examples.

Figure 2: Changes in acid value of carnosic acid soybean oil in application examples.

Figure 3 Changes in carnosic acid content of soybean oil in application examples.

Figure 4 shows the changes in oil and food before and after soybean oil frying in the application example.

Detailed ways

Example 1

S1. Add 25kg of rosemary leaf powder to 250kg of 90% ethanol solution, heat to 70°C for the first extraction for 1 hour. After the extraction is complete, filter out the filtrate, and continue to add 250kg of 90% ethanol solution to the filter residue. Follow the instructions Carry out a second extraction for 1 hour under the previous extraction conditions. After the extraction is completed, filter out the filtrate and combine the two filtrates to form the extraction solution.

S2. Concentrate the extract in step S1 under reduced pressure to 10% of the original volume to obtain a concentrated liquid. The concentrated liquid is then subjected to plate and frame press filtration. After the filter cake is washed with water, the washing liquid and filtrate are combined. The filtrate is obtained, and the washed filter cake is the filter residue.

S3. Extract the filtrate in step S2 with ethyl acetate, concentrate the extract, and dry it under vacuum to obtain crude rosmarinic acid (mass of crude product is 1011.14g, purity is 10.21%).

S4. Add 30 times the equivalent of 60% ethanol solution to the filter residue in step S2 and carry out re-extraction for 1 hour at room temperature. After the re-extraction is completed, filter to obtain the filtrate and filter residue; the filter residue is the crude mixture of ursolic acid and carnosol. Extract (mass of crude extract is 1594.49g, purity of ursolic acid is 25.33%, carnosol is 8.67%).

S5: Use activated carbon to decolorize the filtrate in step S4, then concentrate it into a paste, and dry it under vacuum to obtain crude carnosic acid (crude product is 1510.47g, purity is 39.36%).

Example 2

S1. Add 25kg of rosemary leaf powder to 200kg of 95% ethanol solution, heat to 65°C for the first extraction for 0.5h. After the extraction is complete, filter out the filtrate, and continue to add 200kg of 95% ethanol solution to the filter residue. Carry out a second extraction for 0.5h according to the previous extraction conditions. After the extraction is completed, filter out the filtrate and combine the two filtrates to obtain the extraction solution.

S2. Concentrate the extract in step S1 under reduced pressure to 5% of the original volume to obtain a concentrated liquid. The concentrated liquid is then subjected to plate and frame press filtration. After the filter cake is washed with water, the washing liquid and filtrate are combined. The filtrate is obtained, and the washed filter cake is the filter residue.

S3. Extract the filtrate in step S2 with ethyl acetate, concentrate the extract, and dry it under vacuum to obtain crude rosmarinic acid (mass of crude product is 947.45g, purity is 11.24%).

S4. Add 25 times the equivalent of 55% ethanol solution to the filter residue in step S2 and carry out re-extraction at room temperature for 1.5 hours. After the re-extraction is completed, filter to obtain the filtrate and filter residue; the filter residue is the mixture of ursolic acid and carnosol. Crude extract (the mass of the crude extract is 1987.67g, the purity of ursolic acid is 20.54%, and the purity of carnosol is 9.17%).

S5: Use activated carbon to decolorize the filtrate in step S4, then concentrate it into a paste, and dry it under vacuum to obtain the crude carnosic acid (crude product is 1214.94g, purity is 33.85%).

Example 3

S1. Add 25kg of rosemary leaf powder to 200kg of 85% ethanol solution, heat to 75°C for the first extraction for 1.5 hours. After the extraction is complete, filter out the filtrate, and continue to add 200kg of 85% ethanol solution to the filter residue. Carry out a second extraction for 1.5 hours according to the previous extraction conditions. After the extraction is completed, filter out the filtrate and combine the two filtrates to obtain the extraction solution.

S2. Concentrate the extract in step S1 under reduced pressure to 15% of the original volume to obtain a concentrated liquid. The concentrated liquid is then subjected to plate and frame press filtration. After the filter cake is washed with water, the washing liquid and filtrate are combined. The filtrate is obtained, and the washed filter cake is the filter residue.

S3. Extract the filtrate in step S2 with ethyl acetate, concentrate the extract, and dry it under vacuum to obtain crude rosmarinic acid (mass of crude product is 1429.64g, purity is 8.57%).

S4. Add 35 times the equivalent of 65% ethanol solution to the filter residue in step S2 and carry out re-extraction for 0.5 hours at room temperature. After the re-extraction is completed, filter to obtain the filtrate and filter residue; the filter residue is the mixture of ursolic acid and carnosol. Crude extract (the mass of the crude extract is 1315.32g, the purity of ursolic acid is 31.14%, and the purity of carnosol is 7.94%).

S5: Use activated carbon to decolorize the filtrate in step S4, then concentrate it into a paste, and dry it under vacuum to obtain the crude carnosic acid (crude product is 1650.55g, purity is 36.37%).

Application example 1

Dissolve the crude carnosic acid in Example 1 in 95% ethanol, then add vegetable oil 3.5 times the mass of carnosic acid, stir and mix, vacuum concentrate and remove ethanol and water under reduced pressure, cool and filter to remove precipitate, and finally add new vegetable oil to it to adjust the mass concentration of carnosic acid to 10% to obtain carnosic acid oil.

Antioxidant test test:

(1) Prepare three frying pots, marked A, B and C respectively. Pot A adds 1kg of soybean oil without carnosic acid as a blank control group, pot B and C add final concentrations of 0.05% and 0.1 respectively. 1kg of carnosic acid soybean oil was used as the treatment group.

(2) Take a small amount of oil samples from three pots A, B and C to detect peroxide value, acid value and carnosic acid HPLC content. The results are recorded as 0d as the initial control.

(3) Add equal weight of fried meat products to A, B and C respectively, fry at the same temperature (160°C) for 20 minutes, remove the food, continue heating for 6 hours, and take a small amount of frying oil for peroxidation. value, acid value and carnosic acid HPLC content detection, recorded as 1 day of frying.

The peroxide value and acid value of edible oil were tested using corresponding rapid detection test papers. Both test papers were purchased from Guangdong Dayuan Oasis Food Safety Technology Co., Ltd.

(4) The subsequent frying and testing steps are the same as (3), and the order is recorded as 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 days of frying.

(5) Record in detail the daily frying time and various testing indicators.

1. The antioxidant effect of crude carnosic acid on soybean oil is shown in Table 1 and Figure 1.

Table 1 Determination of peroxide value of carnosic acid soybean oil

As shown in Figure 1, the addition of different specifications of carnosic acid has a significant impact on the antioxidant properties of soybean oil. During the early frying process, it was found that the antioxidant properties of soybean oil did not appear as the frying time increased. However, starting from the fourth day, the soybean oil without adding carnosic acid and containing 0.05% carnosic acid began to show obvious changes, and the peroxide value increased faster after the seventh day, while the soybean oil containing 0.1% carnosic acid The antioxidant properties of soybean oil are stable, indicating that this specification of carnosic acid soybean oil has a significant effect in antioxidant properties.

2. The effect of carnosic acid on the acid value of soybean oil. The test results are shown in Table 2 and Figure 2.

Table 2 Determination of acid value of carnosic acid soybean oil

As shown in Table 2 and Figure 2, the initial acid value of soybean oil added with carnosic acid is higher than that of the control group. As the frying time increases, the acid value of the control group increases significantly. On the 5th day of continuous frying, The acid value of group B containing 0.05% carnosic acid slowed down and reached the limit value on the 11th day of continuous frying, while the performance of group C containing 0.1% carnosic acid was stable, indicating that The 0.1% carnosic acid soybean oil has a significant effect in prolonging the quality change of frying oil.

3. The changes in carnosic acid content during soybean oil frying are shown in Table 3 and Figure 3.

Table 3 Determination of carnosic acid content in soybean oil (HPLC)

As shown in Table 3 and Figure 3, as the frying time increases, the carnosic acid content in soybean oil gradually decreases, indicating that the oxidative deterioration of soybean oil during food frying is related to the change in carnosic acid content. , and carnosic acid can delay the deterioration of frying oil.

4. Effect of carnosic acid antioxidant on sensory evaluation during soybean oil frying

For consumers, the most intuitive source of judgment of food quality is the senses. Quality is generally a subjective judgment of people, which may be affected by different levels of individual feelings. During the frying process of soybean oil, the oil will undergo complex chemical reactions and gradually accumulate a series of deterioration products. These will have adverse effects on the sensory and quality of the oil, which are obvious in the color and viscosity of the oil. It can be seen from Figure 4 that as the frying time increases, the color of the soybean oil in the control group changes significantly before and after frying, and its viscosity and light transmittance also become significantly worse. Compared with the blank control group, under the same frying time , the oil in the antioxidant group has a lighter color and lower viscosity than the control group, and the soybean oil with 0.1% carnosic acid is in better condition, indicating that the addition of carnosic acid can effectively suppress the viscosity of oil during frying. increase and delay the deterioration of oil color, which may be related to the added antioxidants inhibiting the formation of carbonyl compounds in oil.

In terms of the overall results of the antioxidant effect, carnosic acid antioxidant was added to soybean oil, and under high-temperature frying conditions, the effect of the added amount of carnosic acid on the quality of the frying oil was explored. By changing the acid value and process of the oil, The physical and chemical indicators such as oxidation value, color and viscosity were measured to evaluate the practical application effect of carnosic acid antioxidant. The results show that compared with the control group, the addition of carnosic acid can effectively inhibit the increase in the acid value of the frying oil, and at the same time effectively maintain the viscosity and sensory quality of the frying oil; during the first 3 days of frying, the control group The peroxide value of the soybean oil in the experimental group with carnosic acid added was basically unchanged and consistent with that in the experimental treatment group. From 3d to 11d, the rising trend of soybean oil in the experimental group with carnosic acid added slowed down, and the soybean oil with 0.1% carnosic acid added There was basically no change in the oxidation value, while the peroxide value in the control group continued to increase, indicating that the carnosic acid antioxidant had a significant inhibitory effect on the decline in the physical and chemical quality of frying oil, and the most suitable addition specification was 0.1% (HPLC). At the same time, carnosic acid can inhibit the degradation of unsaturated fatty acids in frying oil, which may be related to inhibiting the production of volatile aldehydes in the oil, maintaining the frying performance of the oil, increasing the oxidative stability of the oil, and thus affecting the oil. It plays a protective role in the quality of the frying process.

The above are all preferred embodiments of the present application, and are not intended to limit the scope of protection of the present application. Therefore, any equivalent changes made based on the structure, shape, and principle of the present application shall be covered by the scope of protection of the present application. Inside.

Claims (9)

1. A process for extracting carnosic acid, comprising the following steps: S1. Add rosemary leaves to a mixed solution of ethanol and water with a first-level concentration for heating and extraction. After the extraction is completed, the first-level extraction solution is obtained; S2. Concentrate the first-stage extraction liquid in step S1, perform plate and frame press filtration on the concentrated liquid, and wash the filter cake with water to obtain the filtrate and filter residue; S3. Extract the filtrate in step S2, concentrate the extract, and dry it to obtain crude rosmarinic acid; S4. Re-extract the filter residue in step S2 with a mixed solution of ethanol and water of the second step concentration. After the re-extraction is completed, filter it to obtain the filtrate and filter residue; the filter residue is the crude extraction of ursolic acid and carnosol. thing; S5: Decolorize the filtrate in step S4, and then concentrate it into a paste. After vacuum drying, the crude carnosic acid product can be obtained. 2. The process for extracting carnosic acid according to claim 1, characterized in that, in the step S1, the mass concentration of ethanol in the mixed solution of ethanol and water of the first concentration is 85~95%; the number of extractions 2 to 3 times, the extraction time is 0.5 to 1.5 hours, and the extraction temperature is 65 to 75°C. During each extraction, add a mixed solution that is 8 to 12 times the mass of rosemary. 3. The process for extracting carnosic acid according to claim 1, characterized in that, in the step S2, the concentration adopts vacuum and reduced pressure concentration, and the first-stage extraction liquid is concentrated to 5~15% of the original volume; concentration Ethanol is recovered during the process; the filtrate includes filtered liquid and liquid collected by washing. 4. The process for extracting carnosic acid according to claim 1, characterized in that, preferably, in step S3, the extraction agent is ethyl acetate. 5. The process for extracting carnosic acid according to claim 1, characterized in that, in the step S4, the mass concentration of ethanol in the mixed solution of ethanol and water of the second step concentration is 55~65%; the mixed solvent The added amount is 25~30 times the mass of the filter residue, the re-extraction temperature is room temperature, and the re-extraction time is 0.5~1.5h. 6. The process for extracting carnosic acid according to claim 1, characterized in that, in step S5, activated carbon is used as the decolorizing agent. 7. Extract crude carnosic acid according to the extraction method described in any one of claims 1 to 6. 8. Application of crude carnosic acid according to claim 7 in improving the antioxidant effect in vegetable oils. 9. The application of the crude carnosic acid product in improving the antioxidant effect in vegetable oil according to claim 8, characterized in that, before the crude carnosic acid product is added to the vegetable oil, pretreatment is required, which specifically includes the following steps: Dissolve crude carnosic acid in 90~95% ethanol, then add vegetable oil 3~4 times the mass of carnosic acid, stir and mix, then concentrate under vacuum to remove ethanol and water under reduced pressure, cool and filter to remove precipitates material, and finally add new vegetable oil to it to adjust the mass concentration of carnosic acid to 10% to obtain carnosic acid oil. Carnosic acid oil can be added to vegetable oil as an antioxidant additive to play an antioxidant role.