A flax lignan-icariin compound solid beverage for preventing or adjuvant treating osteoporosis and a preparation method thereof

Abstract

This invention discloses a linseed lignan-icariin compound solid beverage for the prevention or adjuvant treatment of osteoporosis and its preparation method, belonging to the field of functional food processing technology. The main active ingredient is a compound extract of linseed lignan and icariin, supplemented with sorbitol, maltodextrin, and β-cyclodextrin. Flaxseed and epimedium are mixed at a specific mass ratio, extracted using subcritical fluid technology, and purified with macroporous resin to obtain the compound extract; then encapsulated with β-cyclodextrin, and finally mixed with excipients and shaped. This method achieves efficient simultaneous extraction and stabilization of the active ingredients, with a simple process, resulting in a product with good solubility, good taste, and high stability. In vitro, this compound solid beverage effectively promotes osteoblast activity, inhibits osteoclast formation, regulates intestinal flora, and increases the production of beneficial metabolites such as short-chain fatty acids, indicating that it can enhance bone density through multiple pathways and can be used in the preparation of products for the prevention or adjuvant treatment of osteoporosis.

Description

Technical Field

[0001] This invention provides a linseed lignan-icariin compound solid beverage for the prevention or adjuvant treatment of osteoporosis and its preparation method, belonging to the field of functional food processing technology. Background Technology

[0002] Osteoporosis is a systemic metabolic bone disease characterized by decreased bone mass and deterioration of bone microstructure, leading to increased bone fragility and fracture risk. It is common in middle-aged and elderly people. Currently, clinical treatments for osteoporosis mainly include anti-resorption drugs (such as bisphosphonates, estrogens, selective estrogen receptor modulators, and RANKL inhibitors like denosumab) and bone-forming drugs (such as parathyroid hormone analogs). However, long-term use of these chemical drugs is often accompanied by a series of adverse reactions. For example, bisphosphonates may cause esophagitis, gastrointestinal discomfort, rare osteonecrosis of the mandible, and atypical femoral fractures; estrogen therapy may increase the risk of breast cancer, endometrial cancer, and thrombotic events. Therefore, developing safe and effective natural plant-derived alternatives is of significant clinical importance.

[0003] Natural plants and their active ingredients have attracted much attention due to their multi-target, multi-pathway effects and high safety profile. Flaxseed, the mature seed of the flaxseed (Ligustrum lucidum), is rich in alpha-linolenic acid, dietary fiber, protein, and lignans, an important phytoestrogen. Numerous studies have shown that flaxseed lignans and their metabolites possess estrogen-like activity, binding to estrogen receptors on osteoblasts to promote osteoblast proliferation, differentiation, and mineralization, while inhibiting osteoclast activity and reducing bone resorption, thus having a positive effect on bone health. Epimedium, a traditional Chinese medicine for tonifying the kidneys and strengthening yang, has a main active ingredient, icariin, which has been shown to significantly promote osteoblast formation and inhibit osteoclast activity. Icariin can stimulate osteoblast differentiation by activating signaling pathways such as BMP-2 / Smads / Runx2 and Wnt / β-catenin, and inhibit osteoclast formation and function by regulating the OPG / RANKL / RANK system, thereby effectively increasing bone density and improving bone microstructure.

[0004] Although both flaxseed and epimedium have shown potential in improving bone metabolism, research on scientifically combining them to develop convenient functional foods is still limited. Current technologies typically employ separate extraction followed by physical mixing, a process that is cumbersome, costly, and fails to achieve true synergy of active ingredients at the molecular level, thus limiting the full realization of their functional advantages.

[0005] In recent years, compound extraction technology has attracted attention due to its ability to reduce energy consumption and solvent use. However, flaxseed and epimedium have significant differences in physical properties, and their active ingredients differ in polarity and solubility, making efficient simultaneous extraction in the same system a technical challenge. Therefore, developing a key technology that can overcome the differences in raw material properties, achieve efficient synergistic extraction of flaxseed lignans and epimedium glycosides, and ultimately produce a stable and easy-to-use product is of great value for promoting the development of such natural compound functional foods. Summary of the Invention

[0006] To overcome the shortcomings of existing technologies, this invention provides a linoleic acid-icariin compound solid beverage for the prevention or adjuvant treatment of osteoporosis and its preparation method. Specifically, this invention aims to solve the problems of lengthy processes, insufficient synergistic effects of components, and poor stability and solubility of active ingredients in existing separate extraction processes by using innovative compound extraction and β-cyclodextrin inclusion technology. Through detailed in vitro biomimetic digestion experiments, it scientifically verifies the function of improving bone metabolism and regulating intestinal flora through multiple pathways, thereby obtaining a solid beverage product with good taste, excellent stability, and clear bone health support function.

[0007] This invention provides a linolenic acid-icariin compound solid beverage, wherein the beverage uses a compound extract of linolenic acid and icariin as the active ingredient, and the excipients include sorbitol, maltodextrin and β-cyclodextrin.

[0008] In one embodiment of the present invention, the composition of the final solid beverage product, by weight percentage, includes: 7.5-11.5% of a compound extract of linolenic acid and icariin; 4.5-7.5% of sorbitol as a sweetener and humectant to improve taste and prevent product clumping; 2-4% of maltodextrin as a filler and carrier to help improve the solubility and flowability of the product; and the balance being β-cyclodextrin, which serves as both an inclusion material and the main solid matrix.

[0009] In one embodiment of the present invention, the dosage form of the solid beverage is preferably soluble granules, which have superior physical properties: dissolution time is less than 30 seconds; angle of repose is ≤32°, indicating good fluidity; the granules are uniform, the color is natural, the taste is refreshing and slightly sweet, and there is no obvious bitterness or off-flavor.

[0010] In one embodiment of the present invention, the preparation method of the compound extract of linseed lignans and icariin includes the following steps: (1) Compound extraction: After washing and drying flaxseed and epimedium, they are mixed in a mass ratio of 1:1 to 3:1 and pulverized to 40 to 60 mesh. Subcritical butane or propane is used as solvent, and the mass ratio of solvent to mixed raw materials is 10:1. Simultaneous extraction is carried out at a temperature of 40 to 50°C and a pressure of 4 to 5 MPa for 2.5 to 3.5 hours to obtain a compound crude extract. (2) Gradient elution purification: The crude extract was subjected to AB-8 macroporous adsorption resin column chromatography and eluted with ethanol-water gradient (40% ethanol was used to remove impurities and 60%~70% ethanol was used to collect the target component). The collected target eluent was dried to constant weight to obtain a linalool and icariin complex extract with a purity ≥75%.

[0011] This invention provides a method for preparing a compound solid beverage of linolenic acid and icariin, comprising the following steps: (1) Multi-component inclusion: The linolenic acid and icariin complex extract were mixed with β-cyclodextrin at a mass ratio of 1:8 to 1:10 and stirred at 50 to 60°C and 300 to 500 r / min for inclusion. The inclusion rate was ≥90% and the particle size was 1 to 5 μm. (2) Formulation: The inclusion product is mixed with excipients (4.5-7.5% sorbitol, 2-4% maltodextrin, and the remainder β-cyclodextrin) and spray-dried to form a solid beverage.

[0012] Compared with the prior art, the present invention has the following outstanding advantages: (1) Strong process synergy and high efficiency: The subcritical synchronous extraction technology is used for the first time to obtain synergistic components in one step. Compared with the traditional fractionation process, the process and energy consumption are significantly reduced, and cell function experiments have shown that its synergistic effect of promoting bone resorption and inhibiting bone resorption is increased by more than 40%.

[0013] (2) Excellent product performance: Through optimized β-cyclodextrin inclusion technology, the problems of easy degradation of active ingredients and obvious bitterness of icariin have been solved. The product dissolves quickly (<30 s), has good fluidity, and a refreshing taste. The component retention rate is high (≥92%) after 6 months of accelerated testing.

[0014] (3) The efficacy verification system is scientific, complete, and highly predictable: The original in vitro biomimetic digestion and cell model evaluation system overcomes the limitations of traditional single in vitro chemical detection or simple cell experiments. It can not only replace some animal experiments and quickly screen and optimize formulas, but also systematically elucidate the multi-level mechanism of action "from mouth to bone", providing a solid and three-dimensional scientific basis for functional claims. This is a significant improvement of the present invention compared with similar patents.

[0015] (4) The mechanism of action is well explained: The evaluation system of this invention directly confirms that the product has the dual functions of indirect regulation (through the prebiotic effect) and regulation (promoting bone resorption and inhibiting bone resorption) through the "intestinal flora-mediated bone metabolism regulation pathway". The mechanism is clearly explained and the scientific connotation is rich.

[0016] (5) The biomimetic digestion-in vitro colonic fermentation integrated model used in this invention can simulate the complete gastrointestinal digestion and colonic microbiota fermentation process in the human body. Through this model, the release efficiency of active ingredients in digestive fluids can be evaluated, and the promoting effect of fermentation on the proliferation of beneficial bacteria (such as Bifidobacteria) and the production of short-chain fatty acids (especially butyric acid) can be directly monitored, thus providing key in vitro evidence for elucidating the synergistic mechanism of this product from the perspective of "intestinal microbiota-mediated bone metabolism regulation pathway".

[0017] (6) In this invention, flaxseed and epimedium are mixed in a mass ratio of 1:1 to 3:1. Within this range, the two components show the best synergistic effect in promoting bone formation and inhibiting bone resorption. A ratio lower than 1:1 (relative excess of epimedium glycosides) may lead to increased bitterness in the product and a decrease in the synergistic effect; a ratio higher than 3:1 (relative excess of flaxseed lignans) may affect the solubility and taste of the product. Detailed Implementation

[0018] The present invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only for explaining the present invention and are not intended to limit the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Unless otherwise specified in the examples, the procedures should be performed under standard conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0020] Detection method: 1. Component content detection The contents of icariin and linolenic acid in the composite extract obtained during the preparation process of this invention and in the final solid beverage were determined by high performance liquid chromatography.

[0021] 2. In vitro efficacy evaluation methods To scientifically predict the product's bone density enhancement function, this invention adopts the following three-level evaluation system: (1) Bionic digestion experiment: simulate the human gastrointestinal environment, measure the release rate of active ingredients, and evaluate their bioavailability.

[0022] Samples were accurately weighed and subjected to simulated gastric digestion (containing pepsin, pH 2.0, 37℃, shaking for 2 h) and simulated small intestinal digestion (containing pancreatic enzymes and bile salts, pH 6.8, 37℃, shaking for 3 h) sequentially. After digestion, the supernatant was collected by centrifugation, and the contents of icariin and linalool were determined by HPLC. The release rate of each active ingredient was calculated as follows: Release rate of active ingredient (%) = (Content measured in digestion supernatant / Initial theoretical content of the ingredient in the sample) × 100%. A blank control group was also established, i.e., no sample was added, and only an equal volume of digestion solution was used for parallel operation to monitor background interference.

[0023] (2) In vitro colonic fermentation experiment: This experiment uses the above-mentioned biomimetic digestion residue as a substrate to evaluate the product’s regulatory effect on the intestinal microecology, so as to elucidate its potential “intestinal flora-mediated bone metabolism regulation pathway”.

[0024] The residue after biomimetic digestion was used as a fermentation substrate and inoculated with a suspension of gut microbiota from healthy human donors (fresh fecal samples were collected from ethically compliant healthy volunteers who had not used antibiotics, probiotics, prebiotics, or laxatives in the past 3 months; had no history of gastrointestinal diseases, metabolic diseases, autoimmune diseases, or mental illnesses; had regular bowel movements (daily or every other day) with normal fecal characteristics; and whose fecal samples were negative for intestinal pathogens (including pathogenic Escherichia coli, Salmonella, Shigella, parasites, etc.). Under anaerobic conditions, the samples were mixed with pre-reduced sterile phosphate buffer at a weight-to-volume ratio, homogenized, and filtered. The resulting filtrate was used as the fermentation inoculum and had to be used immediately). The samples were then fermented in vitro for 24 hours under anaerobic conditions (37°C, anaerobic environment).

[0025] Gut microbiota analysis: After fermentation, the fermentation broth was collected, and the relative abundance changes of specific beneficial bacteria (such as Bifidobacterium) were analyzed using 16S rRNA gene high-throughput sequencing technology.

[0026] Determination of short-chain fatty acids: After centrifugation and filtration, the content of short-chain fatty acids such as acetic acid, propionic acid, and butyric acid was determined by gas chromatography. The total amount of short-chain fatty acids was calculated as the total amount of short-chain fatty acids (unit: mmol / L).

[0027] A blank control group was set up, in which an equal amount of sterile, inert substrate (such as microcrystalline cellulose) replaced the sample digestion residue, while all other conditions remained the same. The total short-chain fatty acid yield of this blank control group was set as a baseline of 1.0. The relative abundance fold increase of Bifidobacterium and the short-chain fatty acid yield of the sample groups were calculated and compared with the results of this blank control group (set as a multiple of 1.0). It should be noted that, due to the different sources of healthy volunteer fecal samples used in different batches of experiments, the total short-chain fatty acid content of the fermentation blank control group (with sterile, inert substrate replacing the sample) showed a reasonable fluctuation within a certain range (8.0~12.0 mmol / L) between batches. The efficacy evaluation of all sample groups was calculated with the values ​​of the blank control group measured in the same batch as a baseline to ensure the scientific comparability of experimental data.

[0028] (3) Cell function experiments: ① Osteoblast experiment: A mouse pre-osteoblast MC3T3-E1 cell model was used, and cells were co-cultured with samples of different concentrations (processed in biomimetic digestion and sterilized filtration solution). Cell proliferation was detected by CCK-8 assay; early differentiation was detected by alkaline phosphatase staining and quantification (ALP activity); and late calcification was detected by quantifying calcium nodule formation by Alizarin Red S staining.

[0029] ② Osteoclast assay: Using a mouse mononuclear macrophage RAW264.7 cell model, cells differentiated into osteoclasts under RANKL induction. After sample treatment, multinucleated TRAP-positive osteoclasts were counted by tartrate-resistant acid phosphatase staining (TRAP staining); the area of ​​bone resorption lacunae was measured by bone resorption assay.

[0030] A blank control group for cell culture was established, in which cells were cultured in the corresponding basal medium (RANKL for osteoclast experiments) without the addition of sample processing solution. The effects of the sample group on cell proliferation, ALP activity, mineralization area, TRAP-positive cell count, and bone resorption area were calculated based on the "blank control group for cell culture" and the percentage increase or decrease in activity was determined.

[0031] 3. Accelerated Product Stability Testing Methods To predict the product's shelf life and assess its stability, accelerated testing methods were used.

[0032] (1) Test conditions Equipment: Constant temperature and humidity test chamber.

[0033] Temperature and humidity: (40 ± 2) ℃, relative humidity (75 ± 5)%. These conditions are widely used in accelerated testing of food and health products.

[0034] Test period: 6 months in total. Samples were taken and tested at the end of months 0, 1, 2, 3, and 6.

[0035] Sample placement: Place the sealed sample flat in the test chamber to ensure uniform heating and humidity.

[0036] (2) Evaluation indicators and testing methods At each sampling time point, the following indicators were tested on the samples: ① Retention rate of active ingredients: Sample preparation: Accurately weigh 1.0 g of sample and prepare the test solution according to the "Method for Determination of Active Ingredient Content (HPLC Method)".

[0037] Determination and Calculation: The contents of icariin and linalool in the sample were determined by HPLC. The formula for calculating the retention rate of active ingredients is as follows: Retention rate (%) = (Content of a component after accelerated testing / Initial content of the component 0 months before accelerated testing) × 100%; The “Total Retention Rate of Active Ingredients” is calculated as the arithmetic mean of the retention rates of the two components.

[0038] ② Changes in key physical properties: Dissolution time: Measured according to the method described in the invention (1g product / 100mL, 25℃ water), and the time required for complete dissolution was recorded.

[0039] Flowability (angle of repose): determined using the fixed funnel method.

[0040] Sensory evaluation: Observe and record changes in the product's color and odor, as well as whether clumping or deliquescence occurs.

[0041] Microbiological indicators (optional, to enhance persuasiveness): Test for total bacterial count, coliform bacteria, etc., according to the method of GB 4789 National Food Safety Standard to ensure that the product still meets hygiene standards after stability testing.

[0042] (3) Result determination If a product meets all of the following conditions after 6 months of accelerated testing, its stability is considered good: Total retention rate of active ingredients ≥ 90%; The dissolution time should not be extended by more than 20% and should still be less than 40 seconds; There are no unacceptable signs of deterioration such as severe clumping, deliquescence, or off-odors.

[0043] Product Composition and Quantitative Description: In the solid beverage of this invention, the final weight percentage of the compound extract of linolenic acid and icariin follows the following quantitative relationship: Compound extract percentage (%) = [100% - (sorbitol% + maltodextrin%)] / (1 + K), where K is the compound extract and β-carotene extract described in step (4). The inclusion ratio of cyclodextrins (i.e., the denominator value in the range of 1:8 to 1:10). For example, when sorbitol is 6%, maltodextrin is 3%, and the inclusion ratio is 1:9, the proportion of the complex extract is 9.1%. The specific proportion values ​​of each example and comparative example are calculated based on this formula and are noted in the corresponding paragraphs. Based on this proportion and the purity of the active ingredients in the complex extract, the content range of icariin and linolenic acid per gram of the final product can be further calculated.

[0044] Example 1 (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 2:1 and grind them to 50 mesh.

[0045] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3 hours at a temperature of 45℃ and a pressure of 4.5 MPa to obtain the compound crude extract.

[0046] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0047] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a light yellow powdery extract of linalool and icariin.

[0048] (5) Inclusion and formulation: The above-mentioned compound extract (approximately 9.1% by weight of the final product) was mixed with β-cyclodextrin at a mass ratio of 1:9 and included at 55°C and 400 r / min, with an inclusion rate of 92%. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added, and the mixture was spray-dried to form granules.

[0049] Intermediate content determination: HPLC analysis showed that the content of icariin in the compound extract was 28.5%, the content of linalool was 56.7%, the total purity was 85.2%, and the mass ratio of the two was approximately 2:1.

[0050] Final product content description: Based on the formula of this embodiment (compound extract accounts for about 9.1%), the content of icariin in the obtained solid beverage granules is not less than 25 mg / g and the content of linolenic acid is not less than 50 mg / g.

[0051] Product characteristics: The product has good flowability, an angle of repose of 26°, and a dissolution time (25°C water) of 20 seconds; the active ingredient ratio is well-balanced, with excellent encapsulation effect and an encapsulated particle size of 3.5 μm. The product has a harmonious taste, slightly sweet upon entry, without any obvious astringency or bitterness.

[0052] Bionic digestion experiment: Total release rate of active ingredients: icariin 96.2%, linalool 94.8%. After 24 h of in vitro fermentation, the total amount of SCFAs was 48.2 mmol / L, while the blank control group was 10 mmol / L. Butyric acid accounted for 38%, and Bifidobacteria increased 3.1 times (based on the blank control group).

[0053] Cellular function experiments: ALP activity increased by 185%, mineralization area increased by 210%; TRAP-positive cells decreased by 68%, and bone resorption lacunar area decreased by 75%.

[0054] Stability study: The product underwent accelerated testing at 40℃ and 75% RH for 6 months. The total retention rate of active ingredients was ≥94%, the physicochemical properties were stable, and the predicted shelf life at room temperature exceeded 24 months.

[0055] Example 2 (1) Raw material preparation: Mix flaxseed and epimedium in a mass ratio of 1:1 and grind them to 40 mesh.

[0056] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 2.5 hours at a temperature of 40℃ and a pressure of 4 MPa to obtain the compound crude extract.

[0057] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 60% ethanol to obtain the linalool and icariin complex extract.

[0058] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a light yellow powdery extract of linalool and icariin.

[0059] (5) Inclusion and Formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:8 and included at 55°C and 400 r / min, achieving an inclusion rate of 91%. 6% sorbitol, 3% maltodextrin, and the remainder β-cyclodextrin were added, and the mixture was spray-dried to form granules. According to the formulation of this embodiment, the compound extract accounts for approximately 10.1% of the weight of the final solid beverage product. This translates to a content of not less than 35 mg of icariin and not less than 35 mg of linolenic acid per gram of product.

[0060] Intermediate content determination: HPLC analysis showed that the content of icariin in the compound extract was 40.0%, the content of linalool was 41.6%, the total purity was 81.6%, and the mass ratio of the two was approximately 1:1.

[0061] Product characteristics: The product has good flowability, an angle of repose of 28°, a dissolution time (25°C water) of 22 seconds, and an inclusion particle size of 3 μm. The product has a refreshing taste and no unpleasant bitterness.

[0062] Bionic digestion experiment: Release rate of active ingredients: icariin 92.5%, linalool 91.0%. After 24 hours of in vitro fermentation, the total amount of short-chain fatty acids was 38.5 mmol / L, while the blank control group was 10 mmol / L. Butyric acid accounted for 35%, and Bifidobacteria increased by 2.4 times.

[0063] Cellular function experiments: Compared with the blank control, the sample treatment solution increased the alkaline phosphatase activity of osteoblast MC3T3-E1 by 165% and the area of ​​mineralized nodules by 190%; it also reduced the number of TRAP-positive multinucleated cells formed by osteoclast precursor RAW264.7 by 60% and the area of ​​bone resorption lacunae by 65%.

[0064] Stability: The product underwent accelerated testing at 40℃ and 75% RH for 6 months. The total retention rate of active ingredients was ≥90%, and no significant decrease was observed in dissolution time and flowability.

[0065] Example 3 (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 3:1 and grind them to 60 mesh.

[0066] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3.5 hours at a temperature of 50℃ and a pressure of 5.0 MPa to obtain the compound crude extract.

[0067] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 70% ethanol to obtain the complex extract of linalool and icariin.

[0068] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a light yellow powdery extract of linalool and icariin.

[0069] (5) Inclusion and Formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:10 and included at 55°C and 400 r / min, achieving an inclusion rate of 90%. 6% sorbitol, 3% maltodextrin, and the remainder β-cyclodextrin were added, and the mixture was spray-dried to form granules. According to the formulation of this embodiment, the compound extract accounts for approximately 8.3% by weight in the final solid beverage product.

[0070] Intermediate content determination: HPLC analysis showed that the content of icariin in the compound extract was 18.6%, the content of linalool was 53.1%, the total purity was 71.7%, and the mass ratio of the two was approximately 3:1.

[0071] Final product content description: Based on the formula ratio of this embodiment, the content of icariin in the prepared solid beverage granules is not less than 15 mg / g, and the content of linalool is not less than 45 mg / g.

[0072] Product characteristics: The product has good flowability, an angle of repose of 29°, a dissolution time (25°C water) of 28 seconds, and an inclusion particle size of 4.9μm. The product has a slightly sweet taste and an overall balanced flavor.

[0073] Bionic digestion experiment: Total release rate of active ingredients: icariin 93.8%, linalool 92.5%. After 24 h of in vitro fermentation, the total amount of short-chain fatty acids was 36.6 mmol / L, while the blank control group was 8.7 mmol / L at the same time, and Bifidobacteria increased by 2.7 times.

[0074] Cellular function experiments: ALP activity increased by 155%, mineralization area increased by 180%; TRAP-positive cells decreased by 55%, and bone resorption lacunar area decreased by 60%.

[0075] Stability study: The product underwent accelerated testing at 40℃ and 75% RH for 6 months. The total retention rate of active ingredients was ≥91%, the physicochemical properties were stable, and the product performance met expectations.

[0076] Comparative Example 1 (ratio outside the range, mass ratio 0.5:1) (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 0.5:1 and grind them to 50 mesh.

[0077] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3 hours at a temperature of 45℃ and a pressure of 4.5 MPa to obtain the compound crude extract.

[0078] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0079] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a pale yellow powdery compound extract.

[0080] (5) Inclusion and formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:9 and included at 55℃ and 400 r / min, with an inclusion rate of 90%. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added and spray-dried to form granules.

[0081] Intermediate content determination: HPLC analysis showed that the total purity of the obtained compound extract was only 70.4%, with linolenic acid content at 23.7% (purity) and icariin content at 46.7% (purity). The abnormally high purity of individual components indicates an imbalance in the extract composition.

[0082] Product characteristics: Dissolution time extended to 50 seconds, angle of repose increased to 36°. The final product has a noticeably bitter taste and unpleasant mouthfeel.

[0083] Bionic digestion experiment: The steps were exactly the same as in Example 1; the total release rate of active ingredients was: icariin 90.1%, linalool 85.3%. However, after in vitro fermentation, the total amount of short-chain fatty acids was only 22.4 mmol / L, while the blank control group was 12.1 mmol / L. The butyric acid content was 4.5 mmol / L, and the growth rate of Bifidobacteria was only 1.

[0084] Cellular function experiments: ALP activity increased by 45%, mineralization area increased by 50%; TRAP-positive cells decreased by 55%, and bone resorption lacunar area decreased by 50%.

[0085] Stability study: After 6 months of accelerated testing, the product showed significant hygroscopicity and slight clumping, with a total retention rate of approximately 82% for active ingredients.

[0086] Comparative Example 2 (ratio outside the range, mass ratio 4:1) (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 4:1 and grind them to 50 mesh.

[0087] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3 hours at a temperature of 45℃ and a pressure of 4.5 MPa to obtain the compound crude extract.

[0088] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0089] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a pale yellow powdery composite extract. HPLC analysis showed that the content of linalool was 59.3% (purity), the content of icariin was 16.5% (purity), and the total purity was 75.8%.

[0090] (5) Inclusion and formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:9 and included at 55℃ and 400 r / min, with an inclusion rate of 90%. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added and spray-dried to form granules.

[0091] Intermediate content determination: HPLC analysis showed that the total purity of the obtained compound extract was only 75.8%, with linolenic acid content at 59.3% (purity) and icariin content at 16.5% (purity). The abnormally high purity of individual components indicates an imbalance in the extract composition.

[0092] Product characteristics: The final product has poor solubility (>70 seconds), coarse particles, poor flowability (angle of repose 40°), slightly bitter taste, powdery texture, and poor mouthfeel.

[0093] Bionic digestion experiment: The procedure was exactly the same as in Example 1; Total release rate of active ingredients: icariin 88.7%, linalool 90.2%. After in vitro fermentation, the total amount of short-chain fatty acids was 23.1 mmol / L, while the blank control group was 11.5 mmol / L. The butyric acid content was 4.8 mmol / L, and the growth rate of Bifidobacteria was only 0.9 times.

[0094] Cellular function experiments: ALP activity increased by 30%, mineralization area increased by 35%; TRAP-positive cells decreased by 15%, and bone resorption lacunar area decreased by 18%.

[0095] Stability study: During accelerated testing, the product was prone to separation, and its solubility further decreased, with a total retention rate of approximately 80% of the active ingredients.

[0096] Comparative Example 3 (incomplete inclusion ratio, mass ratio 1:12) (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 2:1 and grind them to 50 mesh.

[0097] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3 hours at a temperature of 45℃ and a pressure of 4.5 MPa to obtain the compound crude extract.

[0098] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0099] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a pale yellow powdery compound extract.

[0100] (5) Inclusion and formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:12 and included at 55℃ and 400r / min. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added and spray-dried to form granules.

[0101] Product characteristics: The inclusion rate of the obtained inclusion compound is only 72%, and the particle size distribution is uneven. The dissolution time is 38 seconds, the angle of repose is 34°, the product is slightly cloudy after brewing, the taste is slightly rough, and the aftertaste is slightly bitter.

[0102] Bionic digestion experiment: The steps were exactly the same as in Example 1; due to incomplete inclusion, the total release rate of active ingredients (icariin 85.4%, linalool 83.1%) and the in vitro fermentation effect (total short-chain fatty acids 26.0 mmol / L, compared to 10 mmol / L in the blank control group at the same time, and bifidobacteria growth 1.2 times) were both inferior to those in Example 1.

[0103] Cellular function experiments: ALP activity increased by 105%, mineralization area increased by 120%; TRAP-positive cells decreased by 20%, and bone resorption lacunar area decreased by 22%.

[0104] Stability study: The active ingredient degraded significantly in the accelerated test, and the retention rate was less than 65% after 6 months.

[0105] Comparative Example 4 (excessive inclusion ratio, mass ratio 1:6) (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 2:1 and grind them to 50 mesh.

[0106] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3 hours at a temperature of 45℃ and a pressure of 4.5 MPa to obtain the compound crude extract.

[0107] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0108] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a pale yellow powdery compound extract.

[0109] (5) Inclusion and Formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:6 and included at 55℃ and 400 r / min. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added, and the mixture was spray-dried to form granules.

[0110] Product characteristics: Inclusion rate 82%, poor product flowability (angle of repose 42°), dissolution time <30 seconds, easily absorbs moisture and clumps. The product has an overly strong sweetener taste that masks the natural flavor, and the texture is sticky.

[0111] Bionic digestion experiment: The procedure was exactly the same as in Example 1; the total release rate decreased (icariin 84.0%, linolenic acid 82.5%). After in vitro fermentation, the total amount of short-chain fatty acids was 24.5 mmol / L, while the blank control group was 10 mmol / L, and the Bifidobacteria increased by 1.1 times.

[0112] Cellular function experiments: ALP activity increased by 95%, mineralization area increased by 110%; TRAP-positive cells decreased by 18%, and bone resorption lacunar area decreased by 20%.

[0113] Stability study: The retention rate of active ingredients was approximately 75% after accelerated testing.

[0114] Comparative Example 5 (extraction temperature too low: 35℃) (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 2:1 and grind them to 50 mesh.

[0115] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 2 hours at a temperature of 35℃ and a pressure of 4.0 MPa to obtain the compound crude extract.

[0116] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0117] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a pale yellow powdery compound extract.

[0118] (5) Inclusion and formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:9 and included at 55℃ and 400 r / min. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added and spray-dried to form granules.

[0119] Product characteristics: The yield of the compound extract is approximately 25% lower than that of Example 1, with a total purity of only 65.2%. The color is darker, and trace amounts of degradation products were detected. The extract is dark in color, and degradation products were detected. The dissolution time is 45 seconds, the angle of repose is 33°, the product has a bland flavor, almost no characteristic aroma, and a flat taste.

[0120] Bionic digestion experiment: The steps were exactly the same as in Example 1; the total release rate of active ingredients was acceptable, with icariin release rate of 81.0% and linalool release rate of 79.2%. However, the total amount of short-chain fatty acids after in vitro fermentation was 20.5 mmol / L, while the blank control group was 10 mmol / L. Bifidobacteria growth was only 1-fold, and the fermentation broth had abnormal color.

[0121] Cellular function experiments: ALP activity increased by 70%, mineralization area increased by 80%; TRAP-positive cells decreased by 22%, and bone resorption lacunar area decreased by 25%.

[0122] Stability study: The product initially had a dark color, and degradation products increased during accelerated testing. The retention rate of active ingredients was approximately 78%.

[0123] Comparative Example 6 (Extraction temperature too high: 55℃) (1) Raw material preparation: Mix flaxseed and epimedium at a mass ratio of 2:1 and grind them to 50 mesh.

[0124] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 4 hours at a temperature of 55℃ and a pressure of 4 MPa to obtain the compound crude extract.

[0125] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol to obtain the linalool and icariin complex extract.

[0126] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a pale yellow powdery compound extract.

[0127] (5) Inclusion and formulation: The above-mentioned compound extract was mixed with β-cyclodextrin at a mass ratio of 1:9 and included at 55℃ and 400 r / min. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added and spray-dried to form granules.

[0128] Product characteristics: The extract yield and purity of active ingredients are significantly reduced (approximately 15% lower than in Example 1). Dissolution time is <30 seconds, angle of repose is 31°, the product has a slight burnt smell, and the color is darker, affecting sensory acceptance.

[0129] Bionic digestion experiment: The procedure was exactly the same as in Example 1; the absolute value of the total release rate was low (icariin 86.3%, linolenic acid 85.0%). The in vitro fermentation effect was poor, with a total short-chain fatty acid content of 27.2 mmol / L, compared to 10 mmol / L in the blank control group at the same time. However, the growth of Bifidobacteria was only 1.3 times, and the fermentation broth had an abnormal color.

[0130] Cellular function experiments: ALP activity increased by 88%, mineralization area increased by 100%; TRAP-positive cells decreased by 32%, and bone resorption lacunar area decreased by 35%.

[0131] Stability study: The product initially had a dark color, and degradation products increased during accelerated testing. The retention rate of active ingredients was approximately 77%.

[0132] Comparative Example 7 (Traditional Separate Extraction Process Control) The total mass and proportion of raw materials are the same as in Example 1 (flaxseed: Epimedium = 2:1).

[0133] The extraction process was modified: flaxseed and epimedium were subjected to subcritical extraction and purification according to the conditions of Example 1 to obtain flaxseed lignan and epimedium glycoside extracts. The powders of the two extracts were physically mixed in a ratio (simulating 2:1 in Example 1), and then the mixed powder was mixed with β-cyclodextrin at a mass ratio of 1:9 and incorporated at 55°C and 400 r / min. 6% sorbitol, 3% maltodextrin, and the balance β-cyclodextrin were added, and the mixture was spray-dried to form granules.

[0134] Product characteristics: The processing time is nearly doubled, and the amount of solvent used is increased. The dissolution time is 25 seconds, the angle of repose is 30°, and the product taste is similar to that of Example 1, but the integration and complexity of the flavors are slightly inferior.

[0135] Bionic digestion experiment: The steps were exactly the same as in Example 1; the total release rate of active ingredients (icariin 89.0%, linalool 87.5%) and the in vitro fermentation effect (total short-chain fatty acids 30.1 mmol / L, compared to 10 mmol / L in the blank control group at the same time, and Bifidobacteria growth 1.9 times) were both significantly lower than in Example 1.

[0136] Cellular function experiments: ALP activity increased by 112%, mineralization area increased by 130%; TRAP-positive cells decreased by 35%, and bone resorption lacunar area decreased by 40%.

[0137] Stability study: The chemical stability of the product is comparable to that of Example 1, at approximately 89%, but the production economy and synergistic effect are poor.

[0138] Comparative Example 8 (Single Ingredient Control: Flaxseed Only) Based on the process of Example 1, all the Epimedium in the formulation of Example 1 was replaced with an inert carrier of equal mass (such as microcrystalline cellulose), and only flaxseed was used as raw material, keeping the total amount of feed consistent with that of Example 1.

[0139] (1) Raw material preparation: Mix flaxseed with an inert carrier (such as microcrystalline cellulose) at a mass ratio of 2:1 and grind to 50 mesh. The amount of flaxseed used is the same as that used in Example 1. The inert carrier is used to make up the total amount of material (so that the total mass is equivalent to 3 parts of Example 1). It does not dissolve in the subsequent extraction process and does not interfere with the extraction of active ingredients.

[0140] (2) Composite extraction: Subcritical butane was used as the solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out at 45℃ and 4.5 MPa for 3 hours to obtain the crude extract. After extraction, the inert carrier remained in the waste residue and was removed.

[0141] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol.

[0142] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a light yellow powder of linolenic acid. HPLC analysis showed that the content of linolenic acid in the extract was 89.5%, and icariin was not detected.

[0143] (5) Inclusion and formulation: 9.1% of the above flax lignan extract was taken by weight percentage of the final product and mixed with β-cyclodextrin at a mass ratio of 1:9. The mixture was incorporated at 55℃ and 400 r / min, with an incorporation rate of 90%. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added and spray-dried to form granules.

[0144] Product characteristics: This product is a solid beverage containing only linolenic acid. The dissolution time is 23 seconds, the angle of repose is 29°, and the product has a single, mild flaxseed flavor with a slight sweetness, but the flavor profile is limited.

[0145] Bionic digestion experiment: The procedure was exactly the same as in Example 1; only linseed lignans were released (89.5%). After in vitro fermentation, the total amount of short-chain fatty acids was only 18.1 mmol / L, while the blank control group was 10.8 mmol / L, and the Bifidobacteria increased by 1.1 times.

[0146] Cellular function experiments: ALP activity increased by 95%, mineralization area increased by 110%; TRAP-positive cells decreased by 3%, and bone resorption lacunar area decreased by 5%.

[0147] Stability study: The product was subjected to accelerated testing at 40℃ and RH75% for 6 months. The retention rate of active ingredients was ≥88%, indicating good stability. However, the product has a single function and lacks the synergistic effect of icariin.

[0148] Comparative Example 9 (Single ingredient control: Epimedium only) Based on the process of Example 1, only Epimedium is used as raw material, and all flax seeds in the formulation of Example 1 are replaced with an inert carrier (such as microcrystalline cellulose) of equal mass, keeping the total amount of feed exactly the same as in Example 1.

[0149] (1) Raw material preparation: Mix Epimedium and an inert carrier (such as microcrystalline cellulose) at a mass ratio of 1:2 and pulverize to 50 mesh. The amount of Epimedium used is the same as that in Example 1, and the inert carrier is used to make up the total amount of material.

[0150] (2) Compound extraction: Subcritical butane was used as solvent, with a mass ratio of 10:1 to the mixed raw materials. Simultaneous extraction was carried out for 3 hours at a temperature of 45℃ and a pressure of 4.5 MPa to obtain crude extract. The inert carrier remained in the waste residue.

[0151] (3) Macroporous resin purification: The crude extract was purified by AB-8 resin column. First, impurities were removed by elution with 40% ethanol aqueous solution, and then the target component was collected by elution with 65% ethanol.

[0152] (4) Post-processing: The collected target eluent was dried to constant weight to obtain a light yellow powder of linolenic acid. HPLC analysis showed that the linolenic acid content in the extract was 90.8%, and no linolenic acid was detected.

[0153] (5) Inclusion and formulation: 9.1% of the above-mentioned epimedium extract was mixed with β-cyclodextrin at a mass ratio of 1:9 according to the weight percentage of the final product. The mixture was then included at 55℃ and 400 r / min, with an inclusion rate of 90%. 6% sorbitol, 3% maltodextrin and the balance β-cyclodextrin were added, and the mixture was spray-dried to form granules.

[0154] Product characteristics: The product is a solid beverage containing only icariin, with a dissolution time of 22 seconds and a repose angle of 28°. The product has a typical icariin herbal taste with a prominent bitterness that is difficult to completely neutralize even with sweeteners, resulting in poor palatability.

[0155] Bionic digestion experiment: The procedure was exactly the same as in Example 1; only icariin was released (90.8%). After in vitro fermentation, the total amount of short-chain fatty acids was only 17.8 mmol / L, while the blank control group was 11.7 mmol / L, and the Bifidobacteria increased by 1 time.

[0156] Cellular function experiments: ALP activity increased by 65%, mineralization area increased by 40%; TRAP-positive cells decreased by 55%, and bone resorption lacunar area decreased by 58%.

[0157] Stability study: After 6 months of accelerated testing, the retention rate of active ingredients was ≥87%, indicating good chemical stability of the product, but the taste was poor and the function was limited.

[0158] Tables 1 and 2 summarize the key data of the examples and comparative examples in terms of product characteristics, fermentation characteristics, and stability, highlighting the advantages of the present invention (examples) and the shortcomings of the comparative examples.

[0159] Table 1 Comparison of product characteristics and preparation effects between the examples and comparative examples

[0160] Table 2: Comparison of core efficacy indicators between the examples and comparative examples

[0161] (*Note: The total short-chain fatty acid content of all sample groups is the measured value. The value of the blank control group varies reasonably between batches (8.0~12.0 mmol / L). The ratio of the sample group to the blank control group of the same batch is the multiple shown in the table.) Table Explanation: Examples 1-3: Within the preferred parameter range (raw material ratio 1:1~3:1, extraction temperature 40-50℃, inclusion ratio 1:8~1:10), the product exhibits high inclusion rate, good release rate and beneficial bacteria growth, confirming the effectiveness of the present invention.

[0162] Comparative Examples 1-9: By changing key parameters (such as raw material ratio, inclusion ratio, and extraction temperature), the criticality of the parameters of this invention is highlighted. The comparative examples show that the products are inferior to the examples in terms of taste, stability, and bioavailability.

[0163] In summary, this invention provides a flaxseed-epibriobotry compound solid beverage with advanced technology, clearly defined functions, stable performance, and convenient consumption, as well as its preparation method. This product, through unique compound extraction and inclusion technology, fully leverages the synergistic effects of the two natural active ingredients in enhancing bone density and regulating intestinal health, providing a novel, safe, and effective natural functional food option for the prevention and adjuvant treatment of osteoporosis.

[0164] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention. For example, propane or a mixture of propane and butane can also be used as the subcritical extraction solvent; other types of macroporous resins with similar polarity and pore size, besides AB-8, can also be selected; the solid beverage can be formed by dry granulation, tableting followed by pulverization, etc., in addition to spray drying; erythritol, steviol glycosides, and other permitted varieties can also be used as sweeteners in the excipients. These simple variations or substitutions based on the core concept of the present invention all fall within the scope of protection of the present invention.

Claims

1. A compound solid beverage containing linalool and icariin, characterized in that, The beverage uses a compound extract of linolenic acid and icariin as its active ingredient, and its excipients include sorbitol, maltodextrin and β-cyclodextrin. The preparation method of the linolenic acid and icariin complex extract includes: (1) Compound extraction: After washing and drying flaxseed and Epimedium, the mixture was crushed and extracted with subcritical butane or propane as solvent to obtain a compound crude extract; (2) Gradient elution purification: The crude extract was purified by chromatography through a macroporous adsorption resin column, eluted, and the collected target eluent was dried to constant weight to obtain a compound extract of linalool and icariin.

2. The linalool-icariin compound solid beverage according to claim 1, characterized in that, In step (1), flaxseed and epimedium are mixed in a mass ratio of 1:1 to 3:

1.

3. The linolenic acid-icariin compound solid beverage according to claim 1, characterized in that, In step (1), simultaneous extraction is performed for 2.5 to 3.5 hours at a temperature of 40 to 50°C and a pressure of 4 to 5 MPa.

4. The linalool-icariin compound solid beverage according to claim 1, characterized in that, In step (2), the elution process specifically involves removing impurities with 40% ethanol by volume and collecting the target component with 60% to 70% ethanol by volume.

5. The linolenic acid-icariin compound solid beverage according to claim 1, characterized in that, The composition of the final solid beverage product by weight percentage includes: 7.5-11.5% of linolenic acid and icariin complex extract, 4.5-7.5% of sorbitol, 2-4% of maltodextrin, and the balance being β-cyclodextrin.

6. The linalool-icariin compound solid beverage according to claim 1, characterized in that, In step (2), the macroporous adsorption resin is of type AB-8, type HPD-100, or type D-101.

7. The linalool-icariin compound solid beverage according to claim 1, characterized in that, In step (2), the macroporous adsorption resin is type AB-8.

8. The preparation method of the linalool-icariin compound solid beverage according to any one of claims 1 to 7, characterized in that, Includes the following steps: (1) Multi-component inclusion: The linolenic acid and icariin complex extract was mixed with β-cyclodextrin and stirred at 50~60℃ and 300~500 r / min to obtain the inclusion product with an inclusion rate ≥90% and a particle size of 1~5 μm. (2) Formulation: The inclusion product is mixed with excipients and spray-dried to form a solid beverage.

9. The method according to claim 8, characterized in that, In step (1), the linolenic acid and icariin complex extract is mixed with β-cyclodextrin at a mass ratio of 1:8 to 1:

10.

10. The use of the linolenic acid-icariin compound solid beverage according to any one of claims 1 to 7 in the preparation of functional foods or health foods for enhancing bone density and preventing or adjuvant treatment of osteoporosis.

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