CN107981354A - A kind of preparation method and application of fresh Echinacea oral liquid - Google Patents

Abstract

The invention discloses a preparation method of fresh Echinacea oral liquid, which uses fresh Echinacea purpurea leaves, flowers and roots as raw materials, undergoes high temperature steaming, water beating, ultrasonic enhanced extraction, centrifugal filtration, vacuum degassing, high-pressure homogenization, and canning. The fresh Echinacea purpurea oral liquid was obtained by sealing and ultra-high pressure sterilization process, and the content of cichoric acid in the oral liquid was 0.3-0.6 mg/mL. The invention also discloses the application of the fresh echinacea purpurea oral liquid as a health food for improving immune function. The fresh echinacea purpurea oral liquid of the present invention not only retains the active ingredients of fresh plants to the greatest extent, but also has the characteristics of high dissolution of active ingredients, stable storage and strong immune activity, which is of great significance for the development of health food using fresh plant medicinal materials.

Description

A kind of preparation method and application of fresh echinacea purpurea oral liquid

technical field

The present invention relates to a preparation method and application of fresh echinacea purpurea oral liquid with immune enhancing effect, in particular to a processing method for preserving the active ingredients of fresh herbal medicinal materials, improving the dissolution of active ingredients and slowing down storage degradation, which belongs to the post-harvest processing of agricultural products Or the field of functional health food processing.

Background technique

Echinacea purpurea (L.) Moench is an immunostimulant and immunomodulator that is widely valued internationally. In 2014, the sales of Echinacea purpurea (L.) Moench ranked third in the US herbal dietary supplement market. Its research much attention. The research on the pharmacological active ingredients of Echinacea purpurea found that caffeic acid derivatives (cichoric acid, caffeoyl tartaric acid, chlorogenic acid, caffeic acid, echinacea glycosides, etc.) And so on. Among the caffeic acid derivatives, cichoric acid is one of the most abundant and extremely important immune active ingredients. Therefore, cichoric acid or (and) caffeic acid derivatives (total polyphenols) are one of the important index components for quality control in Echinacea purpurea botanical medicine and its functional health food related to enhancing immune activity.

Zeng Jianguo and others have found that the mass fraction of cichoric acid in fresh Echinacea purpurea is 2.5 to 5.5 times higher than that of dried plants (bud stage: 4.6% vs. 1.7%; full flower stage: 3.5% vs. .0.4%). Many researchers have also found that the content of cichoric acid in dried Echinacea-related products (echinacea tablets, capsules, purple drops, etc.) varies greatly (68.88～242.50mg/100g, Journal of Functional Foods, 2010, 2:77-84; 0.039～ 34.6mg/g, Journal of Agricultural and Food Chemistry, 2003, 51:6922-6933.; 42.37～258.70mg/100g, Journal of Functional Foods, 2010, 2:158-162), which may be related to the drying process of medicinal materials and their Subsequent processing resulted in varying degrees of loss of cichoric acid. Therefore, the content of cichoric acid in health food made from fresh plants without drying is relatively stable, and its immune activity is expected to be improved under the same conditions. However, fresh Echinacea purpurea is perishable, and it contains polyphenol oxidase, which can easily cause the degradation of caffeic acid derivatives such as cichoric acid, the active ingredient. For this reason, researchers have made many attempts on postharvest processing of fresh Echinacea purpurea, for example:

1. Kim HO et al. (Journal of Agricultural of Food Chemistry, 2000, 48:4182-4186) compared the effects of air drying (25°C, 40°C, 70°C), full vacuum microwave drying, and freeze drying on fresh Echinacea chrysanthemum. After the postharvest treatment, it was found that the full-vacuum microwave-dried flowers with low humidity were the same as the freeze-dried flowers, and the cichoric acid content was the highest; 50% of the cichoric acid was retained in the flowers that were air-dried at 25°C, and cichoric acid was retained in the flowers that were air-dried at 70°C. The lowest content, only 20% reserved.

2. Lin SD et al. (Food Chemistry, 2011, 125, 226-231) studied the post-harvest drying methods of fresh Echinacea purpurea roots, stems, leaves, and flowers, and found that the content of cichoric acid or total polyphenols under different processing methods from high to low is vacuum freezing Drying > cold air drying (30°C) > air drying (40°C, 55°C, 70°C). For example, the cichoric acid content of flowers in different drying methods is 38.34mg/g, 33.15mg/g, 22.25mg/g, 14.09mg/g, 4.63mg/g.

3. Zhang YL et al. (Industrial Crops and Products, 2011, 34:873-881) compared water bath blanching (100°C, 10min), high-temperature steam steaming (100°C, 15min), microwave treatment (800W, 300s) This pretreatment + traditional shade-drying method (30°C, 70% RH) was used to dry the roots of fresh Echinacea purpurea for 6 months or 2 years, and compared with the content of cichoric acid in the dried roots of Echinacea purpurea obtained only by the shade-drying method. The content (%) of cichoric acid in the dried medicinal materials obtained by other methods is higher. Take rooting in 2 years as an example: microwave method 2.03%> steaming method 1.97%> blanching method 1.08%> shade drying method 0.83%.

4. Patent CN 100553647C (Application No.: 200610031845.2) discloses a method of cold pressing (the ratio of fresh Echinacea and water is 1:1 to 2) using fresh Echinacea chinensis aerial parts or roots; Echinacea purpurea extract or root juice powder is obtained after drying; the raw material residue is extracted twice with 70-95% ethanol, and the extract is concentrated to obtain the echinacea purpurea extract.

5. Early fresh medicines are generally preserved by squeezing the juice and suspending them in ethanol. For example, 22% ethanol is added to the squeezed juice of 78% fresh Echinacea purpurea as a preservative (Phytomedicine, 2005, 12, 625-631).

6. Wang Yingchao et al. (Food Research and Development, 2015, 36:28-31) extracted cichoric acid in fresh Echinacea purpurea root by microwave synergistic extraction method, and extracted it in 50% ethanol, solid-liquid ratio 1:25 (g/mL), Under the optimal extraction conditions of microwave extraction time 660s, microwave extraction power 300W, and extraction once, the average content of cichoric acid in fresh roots was 158.4μg/g.

To sum up, the current post-harvest processing of fresh Echinacea purpurea is mainly drying or blunt enzyme pretreatment + drying in the shade to make dried medicinal materials, and there are also fresh medicinal materials that are squeezed and suspended in ethanol, or fresh medicinal materials are added with water to extract juice and concentrated to make Echinacea purpurea extract Extracts, or extracts prepared by microwave and alcohol-water extraction of fresh roots, the final products of fresh medicinal materials are mostly ethanol juice or extracts, and the extracts cannot be used directly, and must be supplemented by other pharmacy processing methods, such as making Formed into capsules, oral tablets or formulated into oral liquids, etc.

Echinacea purpurea oral liquid mostly adopts alcoholic water to extract echinacea purpurea (dry) medical material in the market, then through allotment, processing forms. like:

1. Patent CN 102100783 A (application number: 200910255687.2) discloses an immune-enhancing oral liquid with echinacea as the main ingredient, and the formula includes echinacea, astragalus, angelica and tangerine peel. The ethanol extract of Echinacea purpurea and tangerine peel is mixed with the water extract of astragalus and angelica to prepare an oral liquid, which can be used to enhance the immunity of various animals such as pigs, poultry, dogs, and cats.

2. Patent CN 101816694 B (Application No.: 201010185585.0) discloses using dried Echinacea purpurea as raw material, through pulverization→microwave-assisted ethanol extraction→filtering→concentration→filling→sterilization process to obtain Echinacea purpurea oral liquid, and its cichoric acid content is 3- 30mg/mL, can be used to enhance the immune function of chickens and other animals;

3. Patent CN 104257713 A (application number: 201410461283.X) discloses a kind of Echinacea purpurea root medicinal material as raw material, after crushing→50-80% ethanol soaking→ultrasonic extraction→filtering to obtain filtrate 1+filter residue; filter residue is 10-30% Mix % ethanol, ultrasonically extract once → combine the filtrate → concentrate under reduced pressure → add potassium sorbate, adjust the pH to 4-6 → filter → fill → sterilize to obtain Echinacea purpurea root oral liquid, which can significantly enhance chicken Newcastle disease vaccine and pig PRRS The immune effect of vaccines.

4. Patent CN106389498 A (application number: 201611061081.1) discloses a kind of Echinacea purpurea oral liquid, including mixing Echinacea purpurea extract with taurine, sodium benzoate, Tween, sodium hydroxide, ethanol-water, filling nitrogen and sealing it into a can Echinacea purpurea oral liquid is obtained, which can be used for livestock and poultry immunity enhancement and virus prevention and control.

Therefore, it is necessary to develop a complete process for making oral liquid by adding water to the fresh Echinacea purpurea medicinal material. On the one hand, the caffeic acid active ingredients in the fresh medicinal material can be retained and dissolved to the greatest extent; The active ingredient is relatively stable, and the quality of the oral liquid is easy to control, which is convenient for industrialization.

Contents of the invention

The present invention aims to provide a preparation method and application of fresh echinacea oral liquid. Through the method of the present invention, caffeic acid active ingredients in fresh echinacea herbal medicine can be effectively dissolved in the oral liquid, and the active ingredients are relatively stable during storage. The fresh echinacea purpurea oral liquid product processed by the invention has the characteristics of high dissolution of active ingredients, stable and controllable quality, strong health care effect and the like.

The present invention is achieved through the following technical solutions:

A preparation method of fresh echinacea purpurea oral liquid, comprises the steps:

(1) Select fresh, non-verticillium, non-rotten Echinacea purpurea plants, remove silt, wash and drain, divide, slice, and place in high-temperature steam equipment for steaming;

(2) Let each part of the Echinacea purpurea treated by heat steaming in step (1) cool down, place it in a beater or juice extractor, add pure water and polish to obtain a homogenate of Echinacea purpurea;

(3) putting the echinacea purpurea homogenate obtained through step (2) into ultrasonic equipment for auxiliary enhanced extraction;

(4) The Echinacea purpurea homogenate after intensive extraction in step (3) is successively subjected to centrifugal filtration, vacuum degassing, high-pressure homogenization, filling and sealing, and ultra-high pressure sterilization to obtain the fresh Echinacea purpurea oral liquid product, and the finished product needs to be refrigerated or frozen for storage .

Wherein, the fresh Echinacea purpurea plant described in step (1) is harvested at the full bloom stage, and the drained plant is divided into three parts: root, leaf and flower, and its flower and root are cut in a cross shape; Steam treatment at 98-104°C for 10-15 minutes.

Wherein, the mass ratio of Echinacea purpurea root, leaf, flower and pure water described in step (2) is 1:8-12.

Wherein, the ultrasonic-assisted extraction conditions described in step (3) are: ultrasonic power 480-550W, time 28-33min.

Wherein, the sterilization described in the step (4) is a medium-temperature collaborative ultra-high pressure sterilization, the medium temperature is 30-40° C., and the pressure is 280-320 MPa for 12-16 minutes.

The fresh echinacea purpurea oral liquid prepared by the preparation method of the present invention.

The finished product of the fresh Echinacea purpurea oral liquid needs to be refrigerated at 0-4°C or frozen at -18°C;

When no preservatives are added, the finished product can be stored at 0-4°C for 5-6 weeks, and at -18°C for more than 6 months, and the content of active ingredients is relatively stable during storage.

Application of the fresh echinacea purpurea oral liquid in improving immune function health food.

Advantage of the present invention and positive effect are as follows:

1, compared with the traditional Echinacea purpurea oral liquid in the market, the raw materials used in the fresh Echinacea purpurea oral liquid obtained by the present invention are fresh medicinal materials, which contain more active ingredients than dry medicinal materials, and the active ingredients in the oral liquid product have many retentions, high dissolution rate, stable storage, Strong health care effect. In order to achieve this effect, the present invention sequentially slices fresh Echinacea purpurea medicinal materials → steams with high temperature steam → adds water to extract juice → strengthens ultrasonic extraction → filters, degasses, homogenizes, fills, and then sterilizes at medium temperature and high pressure. Among them, on the one hand, high-temperature steaming treatment passivates the polyphenol oxidase in fresh plants, so that it cannot degrade caffeic acid derivatives such as cichoric acid and chlorogenic acid, and improves the retention of active ingredients in fresh medicinal materials; on the other hand, steaming treatment promotes plant The cells are broken, and the active ingredients are more likely to dissolve into the homogenate during the subsequent juicing process. Ultrasonic extraction strengthens the dissolution of active ingredients in the homogenate, so that the active ingredients of fresh medicinal materials can be used efficiently. On the one hand, the medium temperature and ultra-high pressure sterilization treatment can kill the microorganisms in the oral liquid and prolong the shelf life of the Echinacea purpurea oral liquid; on the other hand, the high pressure treatment may further passivate the residual polyphenol oxidase, thereby slowing down the degradation of active ingredients during storage .

2. Fresh echinacea juice (mostly ethanol suspension juice) is generally preserved by fermentation (Biological Agriculture and Horticulture, 2004, 22:133-141) or by adding ethanol or sugar, such as: echinacea ethanol juice (78% or 80%) In general, 20% (22%) ethanol is added as a preservative (Phytomedicine, 2005, 12, 625-631; Archiv Fur Geflugelkunde, 2010, 74, 36-42). Although the ethanol juice of fresh Echinacea purpurea can be preserved, the hydrophilic components are easily lost during storage (Archiv Fur Geflugelkunde, 2010, 74, 36-42). The fresh Echinacea purpurea oral liquid of the present invention is obtained by squeezing and filtering fresh medicinal materials with pure water, and is preserved by necessary sterilization techniques. Without adding preservatives, its refrigerated storage can last for 5-6 weeks, and its frozen storage can last for more than 6 months. If you want to prolong the storage period in refrigerated storage, some preservatives can be added in an appropriate amount. In addition, the loss of the active ingredient (cichoric acid or chlorogenic acid) of the fresh echinacea purpurea oral liquid of the present invention is slowed down during refrigeration or freezing.

Description of drawings

Fig. 1 Echinacea purpurea leaf, flower, root homogenate (1:10) changes in the dissolution rate after ultrasonic enhanced extraction; As can be seen from Fig. 1, chlorogenic acid and cichoric acid in blunt enzyme juice (the homogenate obtained by adding water after steaming treatment) are relatively The dissolution rate of directly squeezed juice is increased by about 2 times; and the dissolution rate of blunt enzyme juice is increased by nearly 20% after ultrasonic treatment;

Before and after the ultrahigh pressure sterilization treatment, the content of Echinacea purpurea leaf, flower and root oral liquid and its content change during cold storage; as can be seen from Figure 2, ultrahigh pressure sterilization is no more than 5% to the loss of cichoric acid content in the oral liquid; cold storage to the 5th Weekly, its cichoric acid content lost about 14%;

Fig. 3 Echinacea purpurea leaf, flower, root oral liquid changes in cichoric acid content during freezing; As can be seen from Fig. 3, when the ultra-high pressure sterilized oral liquid is frozen for 6 months, its content loss is only 10%;

Fig. 4 The effect of Echinacea purpurea oral liquid on delayed hypersensitivity (ear difference) in mice; wherein, *P<0.05, compared with the Control group; #P<0.05, compared with the Positive group. It can be seen from the figure that each administration group (RL, RH, LL, LH, FL, FH) of Echinacea purpurea oral liquid can increase the ear difference of mice (P<0.05), indicating that the T cell immunity of mice after oral administration of Echinacea purpurea oral liquid Function enhancement, and this enhancement effect is better than that of the control drug (P<0.05);

Figure 5 The effect of Echinacea purpurea oral liquid on the phagocytosis index of mice; wherein, *P<0.05, compared with the Control group. It can be seen from the figure that the high-dose groups of Echinacea Purpurea Oral Liquid (RH, LH, FH) can increase the phagocytosis index of mice (P<0.05), indicating that the immune function of monocyte-macrophage in mice is enhanced after taking Echinacea Purpurea Oral Liquid. And this immunoenhancing effect is equivalent to that of the control drug (P>0.05).

Detailed ways

The present invention will be further described in detail below through the specific examples, the following examples are only descriptive, not restrictive, and cannot limit the protection scope of the present invention with this.

Example 1: Determination of cichoric acid and chlorogenic acid content in various parts of fresh Echinacea purpurea

Method A: Determination of cichoric acid and chlorogenic acid in fresh leaves, flowers, and roots of Echinacea purpurea according to USP 36 method with slight improvements. That is, fresh medicinal parts were extracted at 1:25 (g/mL) with 70% ethanol under reflux at 80°C for 15 minutes, and the extract was measured in a high-performance liquid chromatography system. The content was expressed in mg/g based on the weight of fresh medicinal materials.

Method B: Slices of fresh leaves, flowers and roots of Echinacea purpurea, placed in steam at 100 ° C for 10-15 min, let cool, then add 70% ethanol at 1:25 (g/mL) and extract at 80 ° C under reflux for 15 min, take The content of the extract was determined in a high-performance liquid chromatography system, expressed in mg/g, and the results are shown in Table 1.

Table 1 Content of main caffeic acid derivatives in roots, leaves and flowers of Echinacea purpurea (mg/g)

It can be seen from Table 1 that heat steaming at 100°C and inactivating enzyme treatment can effectively prevent the degradation of caffeic acid derivatives such as chlorogenic acid and cichoric acid during the extraction process of fresh Echinacea purpurea (leaves, flowers, roots), and the content of inactivating enzyme treatment is relatively direct The detection is increased by 0.84-1.70 times.

Example 2: Dissolution (rate) of cichoric acid and chlorogenic acid in fresh echinacea purpurea homogenate (1:10) under the action of ultrasonic waves

Echinacea freshly squeezed juice: about 10g of Echinacea root, leaf, and flower each, add 10 times of water to homogenate, and make Echinacea fresh juice (or homogenate).

Echinacea purpurea blunt enzyme juice: about 10g of each part of Echinacea purpurea leaves, flowers, and roots, steamed at 100°C for 10-15 minutes (of which: leaves 10 minutes, roots and flowers 15 minutes) let cool, add 10 times of water to homogenate, and make echinacea purpurea blunt Enzyme juice (or homogenate).

Echinacea purpurea blunt enzyme-ultrasonic juice: put the echinacea purpurea blunt enzyme homogenate in 500W ultrasonic wave for 30 minutes.

Dissolution rate of cichoric acid and chlorogenic acid in each homogenate of Echinacea purpurea: the above homogenate was filtered to obtain clear juice or clarified filtrate, take 2.5mL of filtrate, adjust pH to 3, add 5mL ethyl acetate for extraction, take extract in high-efficiency Liquid chromatographic determination of the dissolution rate (mg/g), and compared with the respective content (mg/g) after the blunt enzyme treatment, the results are represented by the dissolution rate, as shown in Figure 1.

Embodiment 3: preparation of fresh Echinacea purpurea leaf oral liquid

1. Select fresh Echinacea purpurea leaves without verticillium wilt, rinse with clean water and drain, put 150g of fresh echinacea purpurea leaves drained of water into high-temperature steam at 100°C for 10 minutes, and cool slightly;

2. Put the Echinacea purpurea leaves treated in step 1 into a wall-breaking beater, add 10 times the mass of pure water, and beat for 1 minute to obtain uniform and delicate fresh Echinacea purpurea leaf juice;

3. The fresh leaf juice of Echinacea purpurea was subjected to 500W ultrasonic wave for 30 minutes, and the juice was separated through a sieve filter centrifuge after enhanced extraction, and the clarified juice was collected; the clarified juice was vacuum degassed at a vacuum degree of 0.08-0.1MPa for 2 -3min, and high-pressure homogenization under 25MPa once, then canned and sealed, each 50ml; put the packaged Echinacea purpurea juice in a 40℃ ultra-high pressure sterilization equipment, apply a pressure of 300MPa and keep the pressure for 15min, after sterilization, it will be fresh Echinacea purpurea leaf oral liquid finished product. The finished product is refrigerated at 0-4°C or frozen at -18°C, and monitored for microbes and content.

The results of microbiological testing showed that within 5 weeks of refrigeration and within 6 months of freezing, the total number of colonies did not exceed the standard (100cfu/mL), and the mold and yeast also met the beverage hygiene limit standard (<20) (GB7101-2015 standard).

Determination of the content of cichoric acid and chlorogenic acid in Echinacea purpurea oral liquid: take 2.5 mL of the oral liquid, adjust the pH to 3, add 5 mL of ethyl acetate for extraction, and take the extract to determine the content (mg/mL) by high performance liquid chromatography.

The changes in the content of chlorogenic acid and cichoric acid in fresh Echinacea purpurea leaf oral liquid before and after ultra-high pressure sterilization and during storage are shown in Figure 2, and the changes in cichoric acid content during frozen storage are shown in Figure 3.

Embodiment 4: preparation of fresh echinacea chrysanthemum flower oral liquid

1. Select fresh, mildew-free Echinacea flowers, rinse with clean water and drain, cut 150g of Echinacea flowers that have been drained into cross-shaped slices, steam in high-temperature steam at 100°C for 15 minutes, and cool slightly;

2. Put the echinacea leaves treated in step 1 into a wall-breaking beater, add 10 times the mass of pure water, and beat for 2 minutes to obtain uniform and delicate fresh echinacea leaf juice;

3. The echinacea fresh flower juice is subjected to 500W ultrasonic wave for 30 minutes, and the juice is separated through a sieve filter centrifuge after enhanced extraction, and the clarified juice is collected; the clarified juice is vacuum degassed at a vacuum degree of 0.08-0.1MPa for 2- 3min, and high-pressure homogenization under 25MPa once, then canned and sealed, each 50ml; put the packaged echinacea juice in 35℃ ultra-high pressure sterilization equipment, apply 300MPa pressure and keep the pressure for 15min, fresh echinacea can be obtained after sterilization Finished flower oral liquid. The finished product is refrigerated at 0-4°C or frozen at -18°C, and monitored for microbes and content.

The results of microbiological testing showed that within 5 weeks of refrigeration and within 6 months of freezing, the total number of colonies did not exceed the standard (100cfu/mL), and the mold and yeast also met the beverage hygiene limit standard (<20) (GB7101-2015 standard).

The changes in the content of chlorogenic acid and cichoric acid in the fresh Echinacea Chrysanthemum Oral Liquid before and after ultra-high pressure sterilization and during storage are shown in Figure 2, and the changes in the content of cichoric acid during frozen storage are shown in Figure 3.

Embodiment 5: preparation of fresh Echinacea purpurea root oral liquid

1. Select the fresh roots of Echinacea purpurea without mildew and rot, remove the silt and impurities, rinse with water and drain, cut 150g of the fresh roots of Echinacea purpurea in a cross shape, and steam them in high-temperature steam at 100°C for 15 minutes. slightly colder;

2. Put the echinacea leaves treated in step 1 into a wall-breaking beater, add 10 times the mass of pure water, and beat for 2 minutes to obtain uniform and delicate fresh echinacea leaf juice;

3. The echinacea fresh flower juice is subjected to 500W ultrasonic wave for 30 minutes, and the juice is separated through a sieve filter centrifuge after enhanced extraction, and the clarified juice is collected; the clarified juice is vacuum degassed at a vacuum degree of 0.08-0.1MPa for 2- 3min, and high-pressure homogenization under 25MPa once, then canned and sealed, each 50ml; put the packaged Echinacea purpurea juice in an ultra-high pressure sterilization equipment at room temperature 20°C, apply a pressure of 300MPa and keep the pressure for 15min, after sterilization, it will be fresh Echinacea purpurea root oral liquid finished product. The finished product is refrigerated at 0-4°C or frozen at -18°C, and monitored for microbes and content.

The results of microbiological testing showed that within 6 weeks of refrigeration and within 6 months of freezing, the total number of colonies did not exceed the standard (100cfu/mL), and the mold and yeast also met the beverage hygiene limit standard (<20) (GB7101-2015 standard).

The changes in the content of chlorogenic acid and cichoric acid in fresh Echinacea purpurea root oral liquid before and after ultra-high pressure sterilization and during storage are shown in Figure 2, and the changes in cichoric acid content during frozen storage are shown in Figure 3.

Example 6: Effect of Fresh Echinacea Purpurea Oral Liquid on T Cell Delayed Allergy in Mice - Ear Swelling Experiment

5-week-old male Kunming mice were randomly divided into groups according to body weight: normal control group (Control), positive control group (Positive), fresh Echinacea purpurea root, leaf and flower oral liquid administration group (low and high doses). The Control group was administered with normal saline; the Positive group was administered with Nutrilite R Echinacea Healthy Tablets (gastric administration volume: 0.25g/kg/d, the effective dose of cichoric acid was 1.25mg/kg/d. Echinacea Tablets: 0.5g/ tablets, cichoric acid content: 0.5g/100g); fresh Echinacea purpurea leaf oral liquid test group (doses were 1.4mL/kg/d, 3.6mL/kg/d, cichoric acid content: 0.346mg/mL), fresh Echinacea purpurea Flower oral liquid test group (doses were 0.8mL/kg/d, 2.0mL/kg/d, cichoric acid content: 0.626mg/mL), fresh Echinacea purpurea root oral liquid test group (doses were 1.2mL/kg /d, 3.0mL/kg/d; cichoric acid content: 0.399mg/mL), intragastric administration once a day, after 4 weeks of administration, carry out delayed hypersensitivity and carbon clearance experiments respectively. All the determinations were carried out according to the “Functional Evaluation Method for Enhanced Immunity” of health products.

In the delayed-type hypersensitivity experiment, after 4 weeks of administration, the abdominal skin of each mouse was depilated with depilatory cream, with a range of about 3 cm × 3 cm, and 50 μL of dinitrofluorobenzene (DNFB) solution was evenly applied for sensitization. Five days later, 10 μL of DNFB solution was evenly applied to the right ear (both sides) of the mouse for challenge. 24 hours after the challenge, the mice were sacrificed by cervical dislocation, and the left and right ear shells were cut off. Ear pieces with a diameter of 8 mm were removed with a punch and weighed. The results of the weight difference (mg) between the two ears are shown in Figure 4.

Example 7: Detection of the immune function of mouse macrophages by fresh Echinacea purpurea oral liquid - carbon clearance experiment

After 4 weeks of administration, the mice were given 4-fold diluted India ink (dose 0.1mL/10g) through the tail vein, and blood was collected from the orbit of the mice 2min (t ₁ ) and 10min (t ₂ ) after injection, 20μL plasma and 2mL 0.1% Na ₂ CO ₃ solution was mixed, and the absorbance was measured at 600nm. After blood was taken, the mice were sacrificed, and the liver and spleen tissues were weighed. The phagocytosis index α was calculated as follows:

Among them, OD ₁ and OD ₂ are the absorbance at 2 min and 10 min, respectively.

The effect of Echinacea Purpurea Oral Liquid on the phagocytosis index of mice is shown in Figure 5.

Claims (8)

1. a kind of preparation method of fresh Echinacea oral liquid, it is characterised in that include the following steps：

（1）Select it is fresh, wither without Huang, without the Echinacea plant addled, remove silt, cleaning is drained, split, cutting into slices, be placed in High-temperature steam equipment carries out hot steaming processing；

（2）Will be through step（1）Each position of Echinacea that heat steams processing is let cool, and is placed in beater or juice extractor, adds pure water Polishing obtains Echinacea homogenate；

（3）Will be through step（2）Obtained Echinacea homogenate inserts ultrasonic equipment and carries out assisted and strengthened extraction；

（4）By step（3）Echinacea homogenate after strengthening extraction is successively through centrifugal filtration, vacuum outgas, high-pressure homogeneous, filling Sealing, ultrahigh-pressure sterilization, up to fresh Echinacea oral liquid product, finished product need to refrigerate or refrigerated storage.

A kind of 2. preparation method of fresh Echinacea oral liquid according to claim 1, it is characterised in that step（1）Middle institute State fresh Echinacea plant to harvest for full-bloom stage, the plant after draining is divided into root, leaf, flower three parts, it is spent and root is with cross Shape cutting；The heat, which is steamed, is placed in 98-104 DEG C of steam treated 10-15 minutes.

A kind of 3. preparation method of fresh Echinacea oral liquid according to claim 1, it is characterised in that step（2）Middle institute Echinacea root, leaf, flower and quality of pure water ratio are stated as 1:8-12.

A kind of 4. preparation method of fresh Echinacea oral liquid according to claim 1, it is characterised in that step（3）Middle institute The ultrasound-assisted extraction condition stated is：Ultrasonic power 480-550W, time 28-33min.

A kind of 5. preparation method of fresh Echinacea oral liquid according to claim 1, it is characterised in that step（4）Middle institute State sterilization and cooperate with ultrahigh-pressure sterilization for medium temperature, medium temperature is 30-40 DEG C, and pressure 280-320MPa keeps 12-16 min.

6. fresh Echinacea oral liquid made from any preparation methods of claim 1-5.

7. fresh Echinacea oral liquid finished product described in claim 6 need to be in 0-4 DEG C of refrigeration or -18 DEG C of refrigerated storages.

8. application of the fresh Echinacea oral liquid described in claim 6 in terms of immune function health food is improved.