Cornus officinalis compound functional beverage as well as preparation method and application thereof

Abstract

The invention relates to the field of functional foods, in particular to a dogwood composite functional beverage as well as a preparation method and application thereof. The dogwood composite functional beverage is prepared from the following raw materials by mass: 12-15 g of dogwood, 14-16 g of hawthorn, 11-13 g of pericarpium citri reticulatae, 11-13 g of lotus leaves, 0-9 g of radix ophiopogonis, 0-9 g of radix salviae miltiorrhizae, 8-10 g of licorice tablets, 8-10 g of rhizoma phragmitis, 0-5 g of tartary buckwheat, 0-6 g of rhizoma atractylodis macrocephalae, 0-6 g of poria cocos, 0-2 g of abelmoschus manihot and 1-3 g of stevia rebaudiana. The dogwood composite functional beverage prepared by the preparation method disclosed by the invention not only can ensure that the product has a relatively high clarification effect, but also can retain active ingredients to the greatest extent, so that the product has moderate sweetness and fresh and cool taste.

Description

Technical Field

[0001] This invention relates to the field of functional foods, and more particularly to a compound functional beverage made from Cornus officinalis, its preparation method, and its application. Background Technology

[0002] Hyperglycemia is a condition where blood sugar levels rise abnormally beyond the normal range due to a relative or severe deficiency of insulin secretion in the body. Hyperlipidemia, also known as hyperlipidemia or dyslipidemia, refers to elevated levels of triglycerides and total cholesterol in the blood plasma, elevated levels of low-density lipoprotein cholesterol (LDL-C), and decreased levels of high-density lipoprotein cholesterol (HDL-C). The causes of both conditions include gene mutations, various environmental factors, and other diseases. Early symptoms of hyperglycemia and hyperlipidemia are often subtle, and the condition is usually discovered when patients seek medical attention for other illnesses. These two conditions can easily trigger other diseases, such as metabolic neuropathy, atherosclerosis, and acute metabolic crisis.

[0003] Currently, there are numerous blood sugar regulating products on the market. At the same time, the incidence of complications caused by severe diabetes is increasing year by year. However, many blood sugar lowering products on the market overemphasize their efficacy, and the quality of these products varies greatly. Their mechanisms of action are also unclear, which seriously misleads consumers and harms their physical and mental health. Meanwhile, the increasing prevalence of diabetes is seriously affecting the quality of life of the population. Therefore, developing a functional food with certain blood sugar regulating properties has become a market trend. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the purpose of this invention is to provide a Cornus officinalis compound functional beverage, its preparation method and application.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a Cornus officinalis compound functional beverage, which is prepared from raw materials comprising the following quantities: Cornus officinalis 12-15 g, Crataegus pinnatifida 14-16 g, Citrus reticulata peel 11-13 g, Nelumbo nucifera leaf 11-13 g, Ophiopogon japonicus 0-9 g, Salvia miltiorrhiza 0-9 g, Glycyrrhiza uralensis 8-10 g, Phragmites communis rhizome 8-10 g, Tartary buckwheat 0-5 g, Atractylodes macrocephala 0-6 g, Poria cocos 0-6 g, Hibiscus rosa-sinensis 0-2 g, Stevia reticulata 1-3 g.

[0006] Preferably, the Cornus officinalis compound functional beverage is prepared from raw materials comprising the following quantities: Cornus officinalis 15 g, hawthorn 15 g, dried tangerine peel 12 g, lotus leaf 12 g, salvia miltiorrhiza 9 g, ophiopogon japonicus 9 g, licorice root 9 g, reed rhizome 9 g, stevia 2 g. This invention also provides a method for preparing the Cornus officinalis compound functional beverage, comprising the following steps: (1) Place all raw materials in the same container, add water and extract under high pressure to obtain material 1; (2) After centrifuging material 1, mix and stir the supernatant, chitosan solution and water to obtain material 2; (3) After centrifuging material 2, add sucralose, citric acid and xylitol to the supernatant to obtain Cornus officinalis compound functional beverage.

[0007] Preferably, the mass ratio of all raw materials to water in step (1) is 1:8~12.

[0008] Preferably, the high-pressure extraction temperature in step (1) is 100~140℃ and the time is 25~35min.

[0009] Preferably, the volume ratio of the supernatant, chitosan solution and water in step (2) is 2~4 mL: 0~1000 μL: 0~1000 μL; The chitosan solution contains 1-3% chitosan by mass.

[0010] Preferably, the pH value of the supernatant in step (2) is 3.0 to 6.0.

[0011] Preferably, the stirring temperature in step (2) is 20~70℃ and the stirring time is 20~40min.

[0012] Preferably, in step (3), the amount of sucralose added is 0.005 to 0.015% of the supernatant mass, the amount of citric acid added is 0.05 to 0.15% of the supernatant mass, and the amount of xylitol added is 2 to 4% of the supernatant mass.

[0013] This invention also provides an application of Cornus officinalis compound functional beverage in the preparation of hypoglycemic products.

[0014] The present invention has the following technical effects and advantages: This invention determines the optimal combination of Chinese herbal medicines by inhibiting α-glucosidase activity in the extract of different herbal formulas, identifies the optimal extraction method, and uses orthogonal experimental design to determine the clarification process and optimize the sensory quality of the herbal compound beverage. Results show that the high-temperature, high-pressure extraction method yields the highest and most stable extraction rate. Under conditions of 2% chitosan (0.15 g / 100ml), pH 3.0, and a temperature of 50℃, the herbal extract exhibits high permeability and retention of effective components. Flavor is adjusted using 0.005% sucralose, 4% xylitol, and 0.10% citric acid. The resulting Cornus officinalis hypoglycemic functional beverage is characterized by its pleasant sweet and sour taste, moderate color, fragrant aroma, and good mouthfeel, making it an ideal Cornus officinalis compound functional beverage with hypoglycemic effects. Attached Figure Description

[0015] Figure 1 The inhibition rates of different formulations against α-glucosidase; Figure 2 The effect of chitosan addition on the permeability of traditional Chinese medicine extract; Figure 3 The effect of pH on the permeability of traditional Chinese medicine extracts; Figure 4 The effect of temperature on the permeability of traditional Chinese medicine extracts. Detailed Implementation

[0016] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0017] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0018] Example 1: Optimal compatibility study of Cornus officinalis compound functional beverage α-Glucosidase is a digestive enzyme found in the small intestine, responsible for breaking down complex carbohydrates into monosaccharides. Increased α-glucosidase activity accelerates carbohydrate breakdown, leading to elevated blood glucose levels. Inhibiting this enzyme activity is an important strategy for treating hyperglycemia, as it can slow glucose absorption and lower blood sugar. Acarbose, as an α-glucosidase inhibitor, is commonly used to delay sugar absorption and treat hyperglycemia. This example uses acarbose as a positive control to study the inhibition of α-glucosidase activity by different combinations of herbal extracts, aiming to determine the optimal combination for the compound beverage.

[0019] 1. Experimental reagents and equipment traditional Chinese medicine: Cornus officinalis (harvested from Ledao Lake Family Farm, Xingtai City, Hebei Province), hawthorn (batch number: 240802), dried tangerine peel (batch number: 244241102), lotus leaf (batch number: 291240601), licorice (batch number: 272240503), reed rhizome (batch number: 316230803), stevia (batch number: 210602), ophiopogon japonicus (batch number: 119250102), salvia miltiorrhiza (batch number: 257250205), atractylodes macrocephala (batch number: 224250108), poria cocos (batch number: 045250202) (the above medicinal materials were purchased from Tianyu Pharmaceutical), tartary buckwheat, and golden sunflower.

[0020] Experimental equipment: Beakers, reflux condenser, electric ceramic heating furnace, electronic balance, condenser, round-bottom flask, conical flask, pipette, enzyme-linked immunosorbent assay (ELISA) analyzer (SpectraMax M2 from Meigu Molecular Instruments Shanghai Co., Ltd.), rotary evaporator (RE-2000A from Shanghai Yarong Biochemical Instrument Factory), 96-well plate.

[0021] Experimental reagents: p-Nitrophenyl-α-D-glucoside (Shanghai Yuanye Biotechnology Co., Ltd.), acarbose hydrate (Shandong Keyuan Biochemical Co., Ltd.), α-glucosidase (Shanghai Yuanye Biotechnology Co., Ltd.), PBS buffer (Hunan Bikman Holding Co., Ltd.), sodium carbonate solution (Tianjin Hedong District Hongyan Reagent Factory), Wahaha purified water (Hangzhou Wahaha Group Co., Ltd.).

[0022] 2. Compatibility scheme: Formula 1: Cornus officinalis 15 g, hawthorn 15 g, dried tangerine peel 12 g, lotus leaf 12 g, licorice root 9 g, reed rhizome 9 g, tartary buckwheat 5 g, stevia 2 g.

[0023] Formula 2: Cornus officinalis 12 g, hawthorn 15 g, tangerine peel 12 g, lotus leaf 12 g, Ophiopogon japonicus 9 g, licorice root 9 g, reed rhizome 9 g, stevia 2 g.

[0024] Formula 3: Cornus officinalis 15 g, hawthorn 15 g, tangerine peel 12 g, lotus leaf 12 g, salvia miltiorrhiza 9 g, ophiopogon japonicus 9 g, licorice root 9 g, reed rhizome 9 g, stevia 2 g.

[0025] Formula 4: Cornus officinalis 12 g, hawthorn 15 g, tangerine peel 12 g, lotus leaf 12 g, salvia miltiorrhiza 9 g, ophiopogon japonicus 9 g, licorice root 9 g, reed rhizome 9 g, atractylodes macrocephala 6 g, poria cocos 6 g, stevia 2 g.

[0026] Formula 5: Cornus officinalis 12 g, hawthorn 15 g, tangerine peel 12 g, lotus leaf 12 g, salvia miltiorrhiza 9 g, ophiopogon japonicus 9 g, licorice root 9 g, reed rhizome 9 g, golden sunflower 2 g, stevia 2 g.

[0027] 3. Experimental Procedure 3.1 Sample Preparation The drug solution was extracted by heating and reflux with distilled water at a solid-liquid ratio of 1:10. The temperature was set at 100℃ and refluxed for 30 min. After filtration, the filtrate was collected and concentrated under reduced pressure using a rotary evaporator to obtain a drug extract. The extract was dissolved in PBS solution and brought to a final volume of 20 mg / ml to obtain a sample stock solution. The sample stock solution was then diluted with PBS to 0.5 mg / ml, 1 mg / ml, 2 mg / ml, and 4 mg / ml for later use.

[0028] 3.2 Inhibition of α-glucosidase experiment Substrate (pNPG) solution: Accurately weigh pNPG standard and prepare a substrate solution with a concentration of 10 mmol / L (3.01 mg / ml) using PBS buffer, and prepare 2 ml of the solution.

[0029] Reaction termination solution: Weigh 10.8g of 1 mol / L Na2CO3 into a beaker, add an appropriate amount of distilled water to dissolve it, and make up to 100mL. Store at 4℃ for later use.

[0030] Positive control drug acarbose solution: Accurately weigh the acarbose sample, dissolve it in PBS buffer, and prepare acarbose solutions with concentrations of 0.5 mg / ml, 1 mg / ml, 2 mg / ml, and 4 mg / ml, respectively. α-Glucosidase solution: Accurately weigh 0.4 mg (20 U) of 100 U (enzyme activity 50 U / mg) of lyophilized enzyme powder, dissolve it in 0.20 ml of PBS to prepare a stock solution of 100 U / mL. Take the enzyme stock solution and dilute it with PBS buffer to prepare an α-glucosidase solution of 0.2 U / mL for later use.

[0031] The following groups were set up: control blank group, control experimental group, sample blank group, sample experimental group, acarbose blank group, and acarbose experimental group. Control blank group: 80 μL of PBS solution was added; Control experimental group: 40 μL of PBS solution and 40 μL of α-glucosidase solution were added; Acarbose blank group: 40 μL of PBS solution and 40 μL of acarbose were added; Acarbose experimental group: 40 μL of acarbose and 40 μL of α-glucosidase solution were added; Sample blank group: 40 μL of PBS solution and 40 μL of sample solution were added; Sample experimental group: 40 μL of sample solution and 40 μL of α-glucosidase solution were added. After reacting at 37℃ for 20 min, 10 μL of substrate PNPG was added to each group, and the reaction was continued at 37℃ for another 40 min. Finally, 80 μL of Na₂CO₃ was added to each group. After sample addition, the absorbance of each group was measured at 410 nm using an ELISA reader.

[0032] Table 1 Absorbance of different groups After obtaining the absorbance under the microplate reader, the results were plotted in Table 1. The α-glucosidase inhibition rate of acarbose was calculated using the formula: [1-(A1-A2) / (A5-A6)], and the α-glucosidase inhibition rate of the sample to be tested was calculated using the formula: [1-(A3-A4) / (A5-A6)]. The results were then plotted as a line graph.

[0033] 4. Experimental Results The results are as follows Figure 1 As shown, all five formulations exhibited some inhibitory effect on α-glucosidase. Formulation three showed a stable upward trend and reached the inhibitory effect of the acarbose group more quickly, achieving an inhibition rate of 87% at 2 mg / ml and 95% at 4 mg / ml. Formulation three was ultimately selected as the target herbal formula for further experimental research. Specifically, it consists of: Cornus officinalis 15 g, Crataegus pinnatifida 15 g, Citrus reticulata peel 12 g, Nelumbo nucifera leaf 12 g, Salvia miltiorrhiza 9 g, Ophiopogon japonicus 9 g, Glycyrrhiza uralensis 9 g, Phragmites communis rhizome 9 g, and Stevia rebaudiana 2 g.

[0034] Example 2: Study on the clarification process of Cornus officinalis compound functional beverage Chitosan is the only alkaline polysaccharide found in nature. Its molecular chain contains a large number of free amino groups, which carry a positive charge under acidic conditions. Negatively charged colloidal substances in beverages, such as pectin, protein, and tannins, will undergo a charge neutralization reaction with chitosan, forming flocs that then settle, thus achieving clarification. Tannins in Cornus officinalis are the main cause of its astringent taste; using chitosan can effectively reduce this astringency and improve the flavor. Simultaneously, the amino and hydroxyl groups of chitosan can integrate metal ions such as iron and copper, preventing turbidity caused by metal ions, and also slowing down the oxidation of polyphenols, reducing the browning rate of the beverage. Therefore, this embodiment selects chitosan as a clarifying agent to improve the clarity of Cornus officinalis herbal compound beverage.

[0035] 1. Experimental equipment Precision electronic balance, condenser tube, round bottom flask, electric ceramic heating furnace, conical flask, weighing bottle, beaker, induction cooker, medical CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd. KQ-250DE), high-pressure steam sterilizer (Shanghai Shenan Medical Instrument Factory DSX-18L / 280B), Wahaha purified water (Hangzhou Wahaha Group Co., Ltd.), pipette, ear syringe.

[0036] 2. Determination of extraction method for formula three Take 1.5 g of Cornus officinalis, 1.5 g of Crataegus pinnatifida, 1.2 g of Citrus reticulata peel, 1.2 g of Nelumbo nucifera leaf, 0.9 g of Salvia miltiorrhiza, 0.9 g of Ophiopogon japonicus, 0.9 g of Glycyrrhiza uralensis, 0.9 g of Phragmites communis rhizome, and 0.2 g of Stevia reticulata. Mix these ingredients in a 1:10 ratio and divide them into four groups: Group A: decoction method, 100℃, 30 min; Group B: high-pressure extraction method (0.1 MPa), 120℃, 30 min; Group C: reflux method, 100℃, 30 min; Group D: ultrasonic extraction method (40 kHz), 100℃, 30 min. Extract the medicinal liquid using each of these four methods, filter, collect the filtrate, and dilute to 100 ml. Take 5 ml of the diluted solution from each group, place it in a weighing bottle, evaporate to dryness, and obtain the paste. Weigh the paste (see Table 2).

[0037] Table 2. Quality of pastes obtained by different extraction methods Table 2 above compares the four extraction methods. High-pressure extraction (HPIE) has the best extraction efficiency, followed by reflux and decoction methods, while ultrasonic extraction has the worst efficiency. Furthermore, HPIE has a standard deviation of only 0.0025, indicating high stability, more concentrated data, and excellent repeatability. Considering all factors, HPIE is the optimal extraction method among the four.

[0038] 3. Clarification process 3.1 Single-factor design 3.1.1 Effect of chitosan dosage on clarification effect The material was loaded into a container at a ratio of 1:10, and extracted using high-pressure extraction at 120℃ for 30 min. The extract was filtered through gauze to obtain the initial extract. The initial extract was transferred to a 50 ml centrifuge tube and centrifuged at 3000 rpm for 3 min. The supernatant was aliquoted into 10 ml EP tubes, 3 ml per tube. A 2% chitosan solution was added in a gradient of 0 μL, 20 μL, 100 μL, 200 μL, 300 μL, 400 μL, 500 μL, and 1000 μL, respectively. Distilled water was added in the following amounts: 1000 μL, 980 μL, 900 μL, 800 μL, 700 μL, 600 μL, 500 μL, and 0 μL. μL, keeping the total volume consistent, i.e., the final concentrations of chitosan are 0.00g / 100ml (100ml is the total volume of the centrifuged supernatant of the initial extract, 2% chitosan solution and distilled water, the same below), 0.01g / 100ml, 0.05g / 100ml, 0.1g / 100ml, 0.15g / 100ml, 0.2g / 100ml, 0.25g / 100ml and 0.5g / 100ml respectively. Put them into a 40℃ water bath, stir at low speed for 30min, flocculate, cool to room temperature, let stand, centrifuge and take the supernatant, and measure the transmittance at 680 nm.

[0039] 3.1.2 Effect of pH on Clarification The initial extract was transferred to a 50 ml centrifuge tube and placed in a large centrifuge at 3000 rpm for 3 min. After centrifugation, the supernatant was collected, and the pH was adjusted to 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, and 6.0, respectively. The supernatant was then pipetted into 10 ml EP tubes (3 ml per tube), and 400 μL of 2% chitosan solution and 600 μL of distilled water were added. The tubes were placed in a 40℃ water bath and stirred at low speed for 30 min to induce flocculation. After cooling to room temperature and standing, the supernatant was collected by centrifugation, and the transmittance was measured at 680 nm.

[0040] 3.1.3 Effect of Temperature on Clarification Effect The initial extract was transferred to a 50 ml centrifuge tube and centrifuged at 3000 rpm for 3 min. After centrifugation, the supernatant was collected and aliquoted into 10 ml EP tubes (3 ml per tube). 400 μL of 2% chitosan solution and 600 μL of distilled water were added. The mixture was stirred at low speed for 30 min at 20℃, 30℃, 40℃, 50℃, 60℃, and 70℃ respectively to induce flocculation. After cooling to room temperature and standing, the supernatant was collected by centrifugation, and the transmittance was measured at 680 nm.

[0041] 3.2 Orthogonal Design Single-factor experiments were conducted to determine three levels of chitosan dosage, pH, and temperature. A factor level table was established, and a table (Table 6) was drawn using transmittance as an indicator. Orthogonal experiments were used to determine the optimal chitosan dosage, pH, and temperature. Range analysis and multi-factor variance analysis were performed using SPSS software. The absorbance of total flavonoids at 510 nm, chlorogenic acid at 329 nm (20-fold dilution), salvianolic acid B at 286 nm (20-fold dilution), total iridoids at 236 nm (30-fold dilution), and citric acid at 214 nm (30-fold dilution) were measured using a UV spectrophotometer. A table was drawn, and the higher the absorbance, the more target products were represented.

[0042] 4. Results and Analysis 4.1 Effect of chitosan dosage on clarification effect (see Table 3) Table 3 Relationship between chitosan concentration and permeability As shown in Table 3, the permeability increases with increasing chitosan concentration. Within the range of 0.15 g / 100ml to 0.25 g / 100ml, the permeability initially reaches a peak and then decreases. During the overall concentration increase, the low permeability may be due to incomplete impurity adsorption at lower chitosan concentrations. As the chitosan concentration increases, the permeability gradually increases, reaching a maximum of 69.5% at 0.20 g / 100ml. At this point, chitosan molecules effectively adsorb impurities without causing side effects due to excess. The size, density, and settling rate of the flocs reach an optimal balance, and suspended particles in the solution are efficiently removed. However, when the concentration continues to rise, excess chitosan molecules may self-aggregate to form new colloidal particles or undergo secondary adsorption with already settled flocs, leading to an increase in residual microparticles in the solution.

[0043] Furthermore, high concentrations of chitosan may increase the viscosity of the solution, hindering the sedimentation of flocs and even triggering a "restabilization" phenomenon. These factors collectively lead to a decrease in light transmittance, resulting in a worse clarification effect. Therefore, 0.20 g / 100 ml has been determined to be the optimal concentration of chitosan for clarifying traditional Chinese medicine extracts, achieving efficient impurity removal while avoiding the negative effects of excessive addition.

[0044] 4.2 Effect of pH on clarification (see Table 4) Table 4 Relationship between pH and transmittance The initial pH of the stock solution was 3.61. As shown in Table 4, the clarification effect was good at pH 3.0. During the clarification of traditional Chinese medicine extracts using chitosan, the pH value of the solution significantly affected the clarification effect, especially under slightly acidic conditions, where the molecular conformation and adsorption capacity of chitosan exhibited a clear pH dependence. In a slightly acidic environment with a lower pH, the amino groups (-NH2) on the chitosan molecular chain underwent protonation, transforming into positively charged -NH3 groups. + At this point, the chitosan molecular chains fully extend due to electrostatic repulsion, forming a highly open three-dimensional network structure. This extended state significantly increases its contact area with negatively charged impurities (such as proteins, tannins, pectin, etc.) in the herbal extract, thereby enhancing electrostatic adsorption and bridging. Impurity particles rapidly aggregate into large flocs through the "bridging" effect of chitosan molecules, accelerating sedimentation and resulting in a significant reduction in the concentration of suspended particles in the solution, weakened light scattering, and a significant increase in transmittance. After adding sodium bicarbonate solution to adjust the pH of the herbal solution, the transmittance decreases accordingly.

[0045] 4.3 Effect of temperature on clarification (see Table 5) Table 5 Relationship between temperature and transmittance Table 5 shows that when the experimental temperature is controlled within the range of 20℃ to 40℃, the transmittance of the solution changes relatively little. This phenomenon may be mainly related to the flocculation kinetics of chitosan and the complex composition of the drug solution. Within this temperature range, the extension of the chitosan molecular chains is limited. Although the electrostatic adsorption between its amino groups and negatively charged colloidal particles such as proteins and tannins in the extract has been initiated, the molecular thermal motion is relatively slow, and the floc formation rate and sedimentation rate are in a dynamic equilibrium, resulting in a limited increase in transmittance. At the same time, the polysaccharides and mucilage present in the drug solution have high viscosity at low temperatures, which creates certain resistance to the sedimentation of flocs, further limiting the significant improvement in transmittance. When the solution reaches 50℃, the transmittance reaches its peak. At 50℃, the thermal motion of chitosan molecules is enhanced, allowing its molecular chains to fully extend and exposing more active amino sites. At the same time, the viscosity of the traditional Chinese medicine extract is reduced, the resistance to floc sedimentation is reduced, and the clarification process is accelerated. Furthermore, a temperature of 50℃ may have promoted hydrogen bonding or hydrophobic interactions between chitosan and the components of traditional Chinese medicine, enhancing the stability of the flocs and making them easier to settle. However, when the temperature continued to rise above 50℃, the transmittance showed a decreasing trend. This phenomenon may mainly stem from the destructive effect of higher temperatures on the floc structure; the increased thermal motion of the solution may cause the already formed flocs to re-dissociate due to weakened intermolecular forces, leading to the redispersal of some suspended particles in the solution and a decrease in transmittance. Therefore, considering the overall transmittance of the formulation, 50℃ was chosen as a more suitable temperature.

[0046] 4.4 Establish a factor level table Table 6. Three Factors, Three Levels Table 4.5 Results of Orthogonal Experiments Table 7 shows that, based on the single-factor experiments on the clarification effect of chitosan on the drug solution, a comprehensive analysis of the interactions between various factors was conducted, using a three-factor, three-level orthogonal experiment with chitosan dosage, initial pH value, and clarification temperature. The results indicate that the order of influence of the three factors on the clarification effect is: initial pH value > temperature > chitosan dosage. The optimal orthogonal experiment results for clarification effect show that the best combination is a chitosan dosage of 0.25 g / 100 ml, a pH of 3.0, and a temperature of 50℃.

[0047] Table 7 Results of Orthogonal Experiments Based on the absorbance values ​​of the active ingredients (Table 8) and the results analysis (Table 9), the optimal conditions for total flavonoids to achieve an absorbance of 510 nm were: chitosan dosage of 0.15 g / 100 ml, pH 4.0, and temperature of 50℃; for chlorogenic acid to achieve an absorbance of 329 nm, the optimal conditions were: chitosan dosage of 0.25 g / 100 ml, pH 4.0, and temperature of 60℃; for salvianolic acid B to achieve an absorbance of 286 nm, the optimal conditions were: chitosan dosage of 0.25 g / 100 ml, pH 3.5, and temperature of 60℃; and for total iridoids to achieve an absorbance of 236 nm and citric acid to achieve an absorbance of 214 nm, the optimal conditions were: chitosan dosage of 0.15 g / 100 ml, pH 3.0, and temperature of 40℃. Range analysis showed that the chitosan dosage had the greatest impact on the retention of all active ingredients, followed by pH, while temperature had the least impact. However, the analysis of the clarification results showed that the amount of chitosan had a relatively small impact on the clarification. Therefore, considering all factors, the optimal conditions for clarification of the drug solution were selected as follows: chitosan dosage of 0.15 g / 100ml, pH of 3.0, and temperature of 50℃. This ensures both high clarification and maximum retention of the effective components.

[0048] Table 8 Absorbance values ​​of effective components in each group Table 9. Analysis of absorbance values ​​of effective components in each group Example 3: Flavoring Process of Cornus officinalis Compound Functional Beverage Sucralose, a widely used food additive, is an important source of sweetness in the modern food industry. Especially for people with high blood sugar or diabetes, sucralose is a relatively safe sugar substitute, as it is almost not metabolized and absorbed by the body, and does not cause significant fluctuations in blood sugar or insulin levels. Its sweetness intensity is outstanding, approximately 600 times that of sucrose, meaning that even a very small amount can achieve a significant sweetening effect. Based on repeated experiments and sweetness tasting, results show that when sucralose is added to the system at approximately 0.015%, it presents a relatively natural and moderate sweetness, accurately contributing to the base sweetness and avoiding a bitter aftertaste.

[0049] Xylitol, a widely used food additive, has a unique metabolic mechanism that allows it to be metabolized without relying on insulin, making it an ideal sugar substitute for people with high blood sugar and diabetes. Xylitol's sweetness is similar to sucrose, being 0.9 to 1.2 times sweeter, providing a similar sweetness experience while reducing calorie intake. Preliminary experimental results show that when xylitol is added to the system at approximately 4%, it exhibits a mild and sweet quality with a refreshing taste.

[0050] When optimizing the sweetness system of sucralose and xylitol, it is necessary to consider both synergistic effects and sensory balance. Considering the potential nonlinear sweetness enhancement effect that may result from their combination, and to prevent the sweetness from exceeding the threshold and causing a cloying sweetness, this study set the 0.005%-0.015% range of sucralose and the 2%-4% range of xylitol as three levels of parameters in the orthogonal experiment, with 0.015% sucralose and 4% xylitol as the upper limits to ensure sweetness compatibility and process feasibility.

[0051] Therefore, in this embodiment, sucralose, citric acid, and xylitol were used as variable factors. Based on the same total volume of the traditional Chinese medicine extract (the clarified liquid after centrifugation of chitosan and distilled water in Example 2), three different addition levels were selected from each factor to draw a three-factor three-level table (see Table 10) for orthogonal optimization experiments. Sucralose, citric acid, and xylitol were added to the traditional Chinese medicine extract, stirred thoroughly, and range analysis was performed using SPSS software.

[0052] Table 10. Three Factors and Three Waters of Flavored Beverages 1. Scoring Criteria To evaluate the flavor of each blending scheme, a scoring standard was established. Twenty students were selected to conduct a sensory evaluation of the multifunctional beverage. The indicators included color (20 points), texture (10 points), aroma (30 points), and taste (40 points), for a total of 100 points (see Table 11).

[0053] Table 11 Sensory Evaluation Criteria for Cornus officinalis Compound Functional Beverages 2. Results of the orthogonal experiment In the refined exploration of the flavoring process of Cornus officinalis compound functional beverages, this invention systematically analyzed the synergistic effect mechanism of sweeteners and acidulants through a combination of orthogonal experiments and sensory verification. Based on the three-factor, three-level orthogonal experimental design of sucralose (0.005%-0.015%), xylitol (2%-4%), and citric acid (0.05%-0.15%) selected in the preliminary experiments, combined with the sensory scores and range analysis of 9 formulations (see Table 12), the results showed that citric acid had the most significant impact on the overall score (R=4.00). Its acidity regulation not only directly improved the refreshingness of the beverage, but also significantly enhanced the perception of sweetness through taste interaction. Xylitol (R=3.33) was the second most effective, and its low-calorie sweetness characteristics effectively buffered the metallic aftertaste that sucralose might bring while maintaining palatability. Although sucralose (R=2.67) has the highest sweetness ratio, its effect on the overall flavor is relatively mild in the low concentration range (0.005%-0.015%).

[0054] The analysis results showed that the optimal combination was 0.015% sucralose, 4% xylitol, and 0.10% citric acid. However, the flavoring process test results showed that the highest overall score was achieved when the sucralose content was 0.005%, xylitol content was 4%, and citric acid content was 0.10%. To further verify the optimal combination (0.015% sucralose, 4% xylitol, and 0.10% citric acid) obtained from range analysis and the candidate combination with slightly lower sweetness and the highest score (0.005% sucralose, 4% xylitol, and 0.10% citric acid), this invention underwent blind testing. Ten food science students were randomly selected and used a 9-point scale (1 = range, 9 = excellent) to rate the sweet-sour balance, bitterness masking, and refreshingness of the two groups of samples (see Table 13). The results showed that the high-concentration sucralose group (0.015%) had an improved sweetness intensity, but due to the fixed amount of citric acid, some subjects reported that the sweetness was "slightly abrupt." Both groups of samples reduced the bitterness perception threshold through the synergistic effect of sweet and sour, but the low-concentration sucralose group had a milder sweetness and a higher degree of integration with the traditional Chinese medicine base. The high-concentration sucralose group had a longer sweetness residue time, resulting in a lower refreshing score, while the low-concentration group achieved a more refreshing rating due to the rapid taste-clearing effect of citric acid. Therefore, the final choice of 0.005% sucralose, 4% xylitol, and 0.10% citric acid to flavor the Cornus officinalis compound functional beverage yielded a moderately sweet and refreshing taste.

[0055] Table 12 Results of Seasoning Process Tests Table 13 Scoring table for optimal and candidate combinations based on range analysis results Through single-factor experiments, the optimal conditions for clarifying the herbal extract with chitosan were determined by using transmittance. The optimal conditions were 0.15 g / 100 ml of chitosan, pH 3.0, and temperature 50℃. In vitro inhibition of α-glucosidase activity and absorbance detection showed that the herbal extract contained high levels of total flavonoids, iridoids, and some organic acids. These conditions can largely preserve these effective components and ensure the hypoglycemic effect of the herbal extract. The flavor of the extract was adjusted by adding sucralose, citric acid, and xylitol. Sensory evaluation was used during the evaluation process, with several students scoring the beverages at different ratios. The taste, color, and aroma of the beverages were comprehensively considered to ensure the objectivity and accuracy of the evaluation results. Ultimately, it was determined that when the sucralose content was 0.005%, the xylitol content was 4%, and the citric acid content was 0.10%, the product had a suitable sweet and sour taste, moderate color, fragrant aroma, and good taste, making it an ideal compound functional beverage with hypoglycemic effects.

[0056] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A compound functional beverage made from Cornus officinalis, characterized in that, It is prepared from raw materials including the following quantities: Cornus officinalis 12-15 g, Crataegus pinnatifida 14-16 g, Citrus reticulata peel 11-13 g, Nelumbo nucifera leaf 11-13 g, Ophiopogon japonicus 0-9 g, Salvia miltiorrhiza 0-9 g, Glycyrrhiza uralensis 8-10 g, Phragmites communis rhizome 8-10 g, Tartary buckwheat 0-5 g, Atractylodes macrocephala 0-6 g, Poria cocos 0-6 g, Hibiscus rosa-sinensis 0-2 g, Stevia reticulata 1-3 g.

2. The Cornus officinalis compound functional beverage according to claim 1, characterized in that, The Cornus officinalis compound functional beverage is prepared from raw materials including the following quantities: Cornus officinalis 15 g, hawthorn 15 g, dried tangerine peel 12 g, lotus leaf 12 g, salvia miltiorrhiza 9 g, ophiopogon japonicus 9 g, licorice root 9 g, reed rhizome 9 g, stevia 2 g.

3. A method for preparing the Cornus officinalis compound functional beverage according to claim 1 or 2, characterized in that, Includes the following steps: (1) Place all raw materials in the same container, add water and extract under high pressure to obtain material 1; (2) After centrifuging material 1, mix and stir the supernatant, chitosan solution and water to obtain material 2; (3) After centrifuging material 2, add sucralose, citric acid and xylitol to the supernatant to obtain Cornus officinalis compound functional beverage.

4. The preparation method according to claim 3, characterized in that, The mass ratio of all raw materials to water in step (1) is 1:8~12.

5. The preparation method according to claim 3, characterized in that, The high-pressure extraction in step (1) is performed at a temperature of 100~140℃ for 25~35 minutes.

6. The preparation method according to claim 3, characterized in that, The volume ratio of the supernatant, chitosan solution and water in step (2) is 2~4 mL: 0~1000 μL: 0~1000 μL; The chitosan solution contains 1-3% chitosan by mass.

7. The preparation method according to claim 3, characterized in that, The pH value of the supernatant in step (2) is 3.0~6.

0.

8. The preparation method according to claim 3, characterized in that, The stirring temperature in step (2) is 20~70℃ and the stirring time is 20~40min.

9. The preparation method according to claim 3, characterized in that, In step (3), the amount of sucralose added is 0.005~0.015% of the supernatant mass, the amount of citric acid added is 0.05~0.15% of the supernatant mass, and the amount of xylitol added is 2~4% of the supernatant mass.

10. The use of the Cornus officinalis compound functional beverage according to claim 1 or 2, or the Cornus officinalis compound functional beverage prepared by the preparation method according to any one of claims 3 to 9, in the preparation of hypoglycemic products.

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