An oral liquid for aiding digestion, supplementing calcium, iron, and zinc, and its preparation method.

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to an oral liquid for aiding digestion, supplementing calcium, iron, and zinc, and its preparation method. The method includes the following steps: extraction and purification of *Corydalis yanhusuo* polysaccharide; preparation of an auxiliary digestive agent; preparation of an aromatic digestive agent; and formulation of the oral liquid. This invention extracts crude *Corydalis yanhusuo* polysaccharide through water extraction and alcohol precipitation, then performs chromatography using a DEAE cellulose DE-52 column to obtain the *Corydalis yanhusuo* polysaccharide. After acetylation, the *Corydalis yanhusuo* polysaccharide is compounded with a concentrated mixture obtained from the water extraction and concentration of hawthorn, coix seed, and yam. The resulting auxiliary digestive agent, when applied to the oral liquid, can enhance the digestive effect, promote the body's absorption and utilization of the effective components of food, thereby effectively improving and treating indigestion, increasing children's appetite, and preventing malnutrition and decreased immunity caused by indigestion and poor appetite.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an oral liquid for aiding digestion, supplementing calcium, iron, and zinc, and its preparation method. Background Technology

[0002] Children are in a critical stage of growth and development, with a high metabolic rate, but their digestive system is relatively weak and not yet fully developed. In daily life, factors such as irregular eating habits, improper food choices, or spleen and stomach dysfunction can easily lead to indigestion, loss of appetite, and other problems. If these problems are not effectively addressed over a long period, they will not only lead to a decrease in the absorption and utilization of ingested nutrients, resulting in deficiencies in essential trace elements such as calcium, iron, and zinc, as well as various vitamins, causing malnutrition and stunted growth, but will also weaken the body's immune function.

[0003] Oral liquids are an ideal dosage form for children to supplement nutrition and regulate their spleen and stomach due to their accurate dosage, convenient administration, and relatively palatable taste. While some oral liquid products on the market claim to aid digestion or supplement trace elements, they often have limited or insufficient efficacy in aiding digestion, making it difficult to effectively improve stubborn indigestion and poor appetite, and failing to fundamentally address nutrient absorption disorders. Furthermore, some products contain added traditional Chinese medicine ingredients that have unpleasant tastes or residual odors (such as mineral or medicinal tastes), which can easily increase children's resistance to oral liquids, leading to poor compliance and further affecting treatment effectiveness. Summary of the Invention

[0004] To address the aforementioned technical deficiencies, this invention presents an oral liquid for aiding digestion, replenishing calcium, iron, and zinc, and its preparation method. By combining dietary therapy and aromatherapy, the oral liquid's stability is ensured while effectively enhancing its digestive function, thereby improving indigestion and loss of appetite in children.

[0005] A method for preparing an oral liquid that aids digestion, relieves indigestion, and supplements calcium, iron, and zinc includes the following steps:

[0006] S1: Extraction and purification of polysaccharides from cauliflower

[0007] Fresh, disease-free cauliflower seedlings were washed, dried, and crushed before water extraction. The resulting polysaccharide concentrate was purified of protein and then precipitated with alcohol. The crude cauliflower polysaccharide obtained by centrifugation was collected and subjected to chromatography using a DEAE cellulose DE-52 column to obtain cauliflower polysaccharide.

[0008] S2: Preparation of digestive aids

[0009] Dissolve the polysaccharide of *Corydalis yanhusuo* in distilled water, adjust the pH to 9, add acetic anhydride and stir to react, then adjust to neutral and dialyze with deionized water, freeze dry to obtain acetylated *Corydalis yanhusuo* polysaccharide, add acetylated *Corydalis yanhusuo* polysaccharide to a mixed concentrate obtained by decocting and extracting hawthorn, coix seed and yam to obtain an auxiliary digestive agent.

[0010] S3: Preparation of aromatic digestive agents

[0011] The volatile oils of chopped fresh tender Pu-erh tea leaves and fresh patchouli leaves were extracted by steam extraction to obtain a Pu-erh-patchouli mixed volatile oil. Food-grade ammonium bicarbonate and hydroxypropyl-β-cyclodextrin were mixed evenly, and the Pu-erh-patchouli mixed volatile oil was slowly added under the conditions of 55-60℃ and 600-650rpm stirring speed. After the addition was completed, the mixture was centrifuged and dried under reduced pressure to obtain an aromatic digestive agent.

[0012] S4: Compound oral liquid

[0013] Chitosan is added to the digestive aid for flocculation to obtain a clarified digestive aid. Zinc gluconate, calcium gluconate, ferrous gluconate and honey are dissolved in purified water to obtain a calcium, iron and zinc supplement sweetener. The clarified digestive aid, calcium, iron and zinc supplement sweetener, aromatic digestive aid and potassium sorbate are mixed evenly, replenished, filled and sterilized to obtain an oral liquid for digestion, relieving indigestion and supplementing calcium, iron and zinc.

[0014] Further, step S1, the extraction and purification of *Caulis Cinnamomum* polysaccharides, specifically includes the following steps:

[0015] S1.1: After cleaning fresh, disease-free cauliflower seedlings, dry them at 45-50℃ until the moisture content is 5-8%. Then, pulverize them through a 100-110 mesh sieve to obtain cauliflower powder. Mix the cauliflower powder with distilled water at a mass ratio of 1:(30-40), boil at 75-80℃ for 2.5-3 hours, filter, retain the filtrate, and concentrate it under reduced pressure to 20-25% of the original volume to obtain polysaccharide concentrate.

[0016] S1.2: Add n-butanol-chloroform mixed solvent to the polysaccharide concentrate, shake thoroughly for 25-30 min, then centrifuge at 3000-4000 rpm for 10-15 min, take the supernatant and add 3-4 times the volume of anhydrous ethanol, let stand overnight at 4℃, centrifuge to collect the precipitate, and dry at 40-45℃ to obtain crude polysaccharide from cauliflower.

[0017] S1.3: Dissolve the crude polysaccharide of *Caulis Amaranthus* in distilled water to prepare a crude polysaccharide solution with a concentration of 5-6 mg / mL. Then inject the solution into a DEAE cellulose DE-52 column. After eluting the column with distilled water, collect the filtrate, concentrate it to 10-15% of the original volume, and freeze-dry it at -40 to -50℃ to obtain *Caulis Amaranthus* polysaccharide.

[0018] Furthermore, the preparation of the digestive aid in step S2 specifically includes the following steps:

[0019] S2.1: Dissolve the cauliflower polysaccharide in distilled water at a material-to-liquid ratio of 1:(40-45) g / mL, then add NaOH to adjust the pH of the solution to 9, stir for 10-15 min, add 3-5 mL of acetic anhydride dropwise at room temperature while maintaining the pH at 9, stir for 30-35 min after the acetic anhydride has been added, adjust the pH to neutral, add the solution to a dialysis bag and dialyze with deionized water, then take out the solution from the dialysis bag and freeze dry it in a freeze dryer to obtain acetylated cauliflower polysaccharide;

[0020] S2.2: Place hawthorn, coix seed and yam in a container at a mass ratio of 1:(0.5-1):(0.5-1), add 25-30 times the mass of distilled water, and decoct at 80-85℃ for 2-3 hours. Then filter and retain the filtrate, concentrate it to a relative density of 1.08-1.1 to obtain a mixed concentrate. Add 4-5wt% of acetylated cauliflower polysaccharide to the mixed concentrate and stir until the acetylated cauliflower polysaccharide is completely dissolved to obtain an auxiliary digestive agent.

[0021] Furthermore, the preparation of the aromatic digestive agent in step S3 specifically includes the following steps:

[0022] S3.1: Take fresh tender Pu-erh tea leaves and fresh patchouli leaves in a mass ratio of 1:(1-1.2), wash them with water to remove impurities, then air-dry and chop them into strips 0.5-1cm wide. Soak them in distilled water for 10-20 minutes to obtain pre-treated leaves. Place the pre-treated leaves in a volatile oil extraction container and use steam extraction. Steam is introduced from the bottom of the extraction container to dry-steam the pre-treated leaves. Control the temperature in the volatile oil extraction container to 95-100℃. Collect the steam from the pre-treated leaves through a 20-25℃ condenser to obtain distilled liquid. Let the distilled liquid stand until it separates into layers, and collect the upper oily liquid to obtain Pu-erh-patchouli mixed volatile oil.

[0023] S3.2: Mix 2-3 parts by weight of food-grade ammonium bicarbonate and 18-20 parts by weight of hydroxypropyl-β-cyclodextrin evenly, then add 300-350 parts by weight of distilled water at 20-25℃, and stir at 300-350 rpm until clear to obtain an inclusion solution. Under the conditions of 55-60℃ and 600-650 rpm, slowly add 2-3 parts by weight of Pu-erh tea-patchouli mixed volatile oil to the inclusion solution at the same time for 30-35 minutes to obtain a suspension.

[0024] S3.3: Place the suspension in a centrifuge and centrifuge at 4000-5000 rpm for 15-20 min. Collect the precipitate and dry it under reduced pressure at 65-70℃ to obtain the aromatic digestive agent.

[0025] Furthermore, the compounding of the oral solution in step S4 specifically includes the following steps:

[0026] S4.1: Add chitosan to the digestive aid and stir evenly. After standing and flocculating, filter with 600-mesh filter paper to obtain a clear digestive aid.

[0027] S4.2: Dissolve 0.08-0.1 parts by weight of zinc gluconate, 0.08-0.1 parts by weight of calcium gluconate, 0.08-0.1 parts by weight of ferrous gluconate and 1-2 parts by weight of honey in 3-5 parts by weight of purified water to obtain a calcium, iron and zinc supplement sweetener;

[0028] S4.3: Mix 30-42 parts by weight of clarifying digestive aid, 24-30 parts by weight of calcium, iron and zinc supplement sweetener, 4-6 parts by weight of aromatic digestive aid and 0.08-0.1 parts by weight of potassium sorbate evenly, add purified water to make up to 100 parts by weight, then fill into 10 mL vials and sterilize in a 115℃ sterilizer for 30 min to obtain an oral liquid for digestion, calcium, iron and zinc supplementation.

[0029] Furthermore, the n-butanol-chloroform mixed solvent in step S1.2 is prepared by mixing n-butanol and chloroform in a volume ratio of 1:5, and the added n-butanol-chloroform mixed solvent accounts for 20-25% of the volume of the polysaccharide concentrate.

[0030] Furthermore, in step S2.1, the molecular weight cutoff of the dialysis bag is 8000-10000 Da.

[0031] Furthermore, the dialysis time in step S2.1 is 2-3 days, and the deionized water is changed every 8-12 hours.

[0032] Furthermore, in step S4.1, the amount of chitosan added accounts for 2-3 wt% of the auxiliary digestive agent, the flocculation temperature is 30-35℃, and the flocculation time is 4-5 h.

[0033] An oral liquid for aiding digestion, relieving indigestion, and supplementing calcium, iron, and zinc is prepared by the above-mentioned method for preparing an oral liquid for aiding digestion, relieving indigestion, and supplementing calcium, iron, and zinc.

[0034] The beneficial effects are as follows: 1. This invention extracts crude polysaccharide from *Corydalis yanhusuo* through water extraction and alcohol precipitation, and then performs chromatography on it using a DEAE cellulose DE-52 column to obtain *Corydalis yanhusuo* polysaccharide. After acetylation of the *Corydalis yanhusuo* polysaccharide, acetylated *Corydalis yanhusuo* polysaccharide is obtained, which can promote the proliferation of beneficial bacteria in the human intestine and inhibit the reproduction of harmful bacteria. When combined with a mixed concentrate obtained from the water extraction and concentration of hawthorn, coix seed and yam, the resulting auxiliary digestive agent can be used in oral liquid to enhance the digestive and relieving effects, promote the body's absorption and utilization of the effective components of food, thereby effectively improving and treating indigestion, improving children's appetite, and preventing malnutrition and decreased immunity caused by indigestion and poor appetite.

[0035] 2. This application uses steam extraction to co-extract volatile oil components from fresh tender Pu-erh tea leaves and fresh patchouli leaves to obtain a Pu-erh-patchouli mixed volatile oil. This Pu-erh-patchouli mixed volatile oil is then formulated into an aromatic digestive aid and added to an oral liquid. The fresh woody and ripe fruity aroma of the Pu-erh volatile oil, combined with the woody, herbal, and spicy aroma of the patchouli volatile oil, creates different layers of fragrance. After taking the oral liquid, which aids digestion, relieves stagnation, and supplements calcium, iron, and zinc, the residual aroma effectively invigorates the spleen and improves digestion and appetite in children.

[0036] 3. In this application, ammonium bicarbonate and hydroxypropyl-β-cyclodextrin are mixed and used to encapsulate the volatile oil of Pu-erh tea and patchouli. During the encapsulation process of hydroxypropyl-β-cyclodextrin with the volatile oil of Pu-erh tea and patchouli, ammonium bicarbonate continuously decomposes at a temperature of 55-60℃ to generate microbubbles, creating numerous microchannels on the wall material of hydroxypropyl-β-cyclodextrin. These microchannels allow the gas molecules of the volatile oil of Pu-erh tea and patchouli to dissipate while ensuring that the oil cannot pass through. This prevents the volatile oil from seeping into the oral liquid, causing stratification and a deterioration in taste. At the same time, it allows the special aroma of the volatile oil of Pu-erh tea and patchouli to dissipate, preventing the taste of the oral liquid from affecting the palate and causing further loss of appetite in children. Detailed Implementation

[0037] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. 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.

[0038] Example 1

[0039] A method for preparing an oral liquid that aids digestion, relieves indigestion, and supplements calcium, iron, and zinc includes the following steps:

[0040] S1: Extraction and purification of polysaccharides from cauliflower

[0041] S1.1: After cleaning fresh, disease-free cauliflower seedlings, dry them at 45℃ until the moisture content is 5%. Then, crush them and sieve them through a 100-mesh screen to obtain cauliflower powder. Mix the cauliflower powder with distilled water at a mass ratio of 1:30. Boil at 75℃ for 2.5 hours and then filter. Keep the filtrate and concentrate it under reduced pressure to 20% of the original volume to obtain polysaccharide concentrate.

[0042] S1.2: Add n-butanol-chloroform mixed solvent to the polysaccharide concentrate. The n-butanol-chloroform mixed solvent is prepared by mixing n-butanol and chloroform in a volume ratio of 1:5. The added n-butanol-chloroform mixed solvent accounts for 20% of the volume of the polysaccharide concentrate. Shake thoroughly for 25 min, then centrifuge at 3000 rpm for 10 min. Take the supernatant and repeat the operation until no protein precipitation occurs. Add 3 times the volume of anhydrous ethanol to the final supernatant, let stand overnight at 4℃, centrifuge to collect the precipitate, and dry at 40℃ to obtain cauliflower crude polysaccharide.

[0043] S1.3: Dissolve the crude polysaccharide of *Caulis Amaranthus* in distilled water to prepare a crude polysaccharide solution with a concentration of 5 mg / mL. Then inject the solution into a DEAE cellulose DE-52 column. After eluting the column with distilled water, collect the filtrate, concentrate it to 10% of the original volume, and freeze-dry it at -40℃ to obtain *Caulis Amaranthus* polysaccharide.

[0044] S2: Preparation of digestive aids

[0045] S2.1: Dissolve cauliflower polysaccharide in distilled water at a material-to-liquid ratio of 1:40 g / mL, then add NaOH to adjust the pH of the solution to 9, stir for 10 min, add 3 mL of acetic anhydride dropwise at room temperature while maintaining the pH at 9, stir for 30 min after the acetic anhydride is added, adjust the pH to neutral, and then add it to a dialysis bag with a molecular weight cutoff of 8000 Da for dialysis with deionized water for 2 days. Change the deionized water every 8 hours of dialysis. Then take out the solution from the dialysis bag and freeze dry it in a freeze dryer to obtain acetylated cauliflower polysaccharide.

[0046] S2.2: Hawthorn, coix seed and yam are placed in a container in a mass ratio of 1:0.5:0.5, and 25 times the mass of distilled water is added. The mixture is decocted at 80℃ for 2 hours, then filtered and the filtrate is retained. The filtrate is concentrated to a relative density of 1.08 to obtain a mixed concentrate. 4 wt% of acetylated cauliflower polysaccharide is added to the mixed concentrate and stirred until the acetylated cauliflower polysaccharide is completely dissolved to obtain an auxiliary digestive agent.

[0047] S3: Preparation of aromatic digestive agents

[0048] S3.1: Take fresh tender Pu-erh tea leaves and fresh patchouli leaves in a mass ratio of 1:1. After washing to remove impurities, dry them in the shade and cut them into strips with a width of 0.5cm. Then soak them in distilled water for 10 minutes to obtain pre-treated leaves. Place the pre-treated leaves in a volatile oil extraction container and use the steam extraction method. Steam is introduced from the bottom of the extraction container to dry-steam the pre-treated leaves. The temperature in the volatile oil extraction container is controlled at 95℃. The steam passing through the pre-treated leaves is collected through a 20℃ condenser to obtain distilled liquid. Let the distilled liquid stand until it separates into layers and collect the upper oily liquid to obtain Pu-erh-patchouli mixed volatile oil.

[0049] S3.2: Mix 2 parts by weight of food-grade ammonium bicarbonate and 18 parts by weight of hydroxypropyl-β-cyclodextrin evenly, then add 300 parts by weight of distilled water at 20°C, and stir at 300 rpm until clear to obtain an inclusion solution. Under the condition of 55°C and 600 rpm stirring speed, slowly add 2 parts by weight of Pu-erh tea-patchouli mixed volatile oil to the inclusion solution at the same time for 30 min to obtain a suspension.

[0050] S3.3: Place the suspension in a centrifuge and centrifuge at 4000 rpm for 15 min. Collect the precipitate and dry it under reduced pressure at 65℃ to obtain the aromatic digestive agent.

[0051] S4: Compound oral liquid

[0052] S4.1: Add chitosan to the digestive aid, stir evenly, and allow it to stand and flocculate. The amount of chitosan added is 2wt% of the digestive aid. The flocculation temperature is 30℃ and the flocculation time is 4h. Then filter it with 600-mesh plate and frame filter paper to obtain a clear digestive aid.

[0053] S4.2: Dissolve 0.08 parts by weight of zinc gluconate, 0.08 parts by weight of calcium gluconate, 0.08 parts by weight of ferrous gluconate and 1 part by weight of honey in 3 parts by weight of purified water to obtain a calcium, iron and zinc supplement sweetener;

[0054] S4.3: Mix 30 parts by weight of clarifying digestive aid, 24 parts by weight of calcium, iron and zinc supplement sweetener, 4 parts by weight of aromatic digestive aid and 0.1 parts by weight of potassium sorbate evenly, add purified water to make up to 100 parts by weight, then fill into 10 mL vials and sterilize in a 115℃ sterilizer for 30 min to obtain an oral liquid for digestion, calcium, iron and zinc supplementation.

[0055] Example 2

[0056] A method for preparing an oral liquid that aids digestion, relieves indigestion, and supplements calcium, iron, and zinc includes the following steps:

[0057] S1: Extraction and purification of polysaccharides from cauliflower

[0058] S1.1: After cleaning fresh, disease-free cauliflower seedlings, dry them at 48℃ until the moisture content is 6%. Then, crush them and sieve them through a 110-mesh screen to obtain cauliflower powder. Mix the cauliflower powder with distilled water at a mass ratio of 1:35. Boil at 78℃ for 2.8 hours and then filter. Keep the filtrate and concentrate it under reduced pressure to 23% of the original volume to obtain polysaccharide concentrate.

[0059] S1.2: Add n-butanol-chloroform mixed solvent to the polysaccharide concentrate. The n-butanol-chloroform mixed solvent is prepared by mixing n-butanol and chloroform in a volume ratio of 1:5. The added n-butanol-chloroform mixed solvent accounts for 22% of the volume of the polysaccharide concentrate. Shake thoroughly for 28 min, then centrifuge at 3500 rpm for 12 min. Take the supernatant and repeat the operation until no protein precipitation occurs. Add 3.5 times the volume of anhydrous ethanol to the final supernatant, let stand overnight at 4℃, centrifuge to collect the precipitate, and dry at 42℃ to obtain the crude polysaccharide of cauliflower.

[0060] S1.3: Dissolve the crude polysaccharide of *Caulis Amaranthus* in distilled water to prepare a crude polysaccharide solution with a concentration of 5.5 mg / mL. Then inject the solution into a DEAE cellulose DE-52 column. After eluting the column with distilled water, collect the filtrate, concentrate it to 12% of the original volume, and freeze-dry it at -45℃ to obtain *Caulis Amaranthus* polysaccharide.

[0061] S2: Preparation of digestive aids

[0062] S2.1: Dissolve cauliflower polysaccharide in distilled water at a material-to-liquid ratio of 1:43 g / mL, then add NaOH to adjust the pH of the solution to 9, stir for 12 min, add 4 mL of acetic anhydride dropwise at room temperature while maintaining the pH at 9, stir for 33 min after the acetic anhydride is added, adjust the pH to neutral, and then add it to a dialysis bag with a molecular weight cutoff of 9000 Da for dialysis with deionized water for 2.5 days. Change the deionized water every 10 h of dialysis. Then take out the solution from the dialysis bag and freeze dry it in a freeze dryer to obtain acetylated cauliflower polysaccharide.

[0063] S2.2: Hawthorn, coix seed and yam are placed in a container in a mass ratio of 1:0.8:0.8, and 27 times the mass of distilled water is added. The mixture is decocted at 82℃ for 3 hours, then filtered and the filtrate is retained. The filtrate is concentrated to a relative density of 1.09 to obtain a mixed concentrate. 4.5wt% of acetylated cauliflower polysaccharide is added to the mixed concentrate and stirred until the acetylated cauliflower polysaccharide is completely dissolved to obtain an auxiliary digestive agent.

[0064] S3: Preparation of aromatic digestive agents

[0065] S3.1: Take fresh tender Pu-erh tea leaves and fresh patchouli leaves in a mass ratio of 1:1.1. After washing to remove impurities, dry them in the shade and cut them into strips with a width of 0.8cm. Then soak them in distilled water for 15 minutes to obtain pre-treated leaves. Place the pre-treated leaves in a volatile oil extraction container and use the steam extraction method. Steam is introduced from the bottom of the extraction container to dry-steam the pre-treated leaves. The temperature in the volatile oil extraction container is controlled at 97℃. The steam passing through the pre-treated leaves is collected through a 23℃ condenser to obtain distilled liquid. Let the distilled liquid stand until it separates into layers and collect the upper oily liquid to obtain Pu-erh-patchouli mixed volatile oil.

[0066] S3.2: Mix 2.5 parts by weight of food-grade ammonium bicarbonate and 19 parts by weight of hydroxypropyl-β-cyclodextrin evenly, then add 325 parts by weight of distilled water at 23°C, and stir at 325 rpm until clear to obtain an inclusion solution. Under the conditions of 58°C and 625 rpm stirring speed, slowly add 2.5 parts by weight of Pu-erh tea-patchouli mixed volatile oil to the inclusion solution at the same time for 33 min to obtain a suspension.

[0067] S3.3: Place the suspension in a centrifuge and centrifuge at 4500 rpm for 17 min. Collect the precipitate and dry it under reduced pressure at 68℃ to obtain the aromatic digestive agent.

[0068] S4: Compound oral liquid

[0069] S4.1: Add chitosan to the digestive aid, stir evenly, and allow it to stand and flocculate. The amount of chitosan added is 2.5 wt% of the digestive aid. The flocculation temperature is 32℃ and the flocculation time is 4.5 h. Then filter it with 600 mesh plate and frame filter paper to obtain a clear digestive aid.

[0070] S4.2: Dissolve 0.09 parts by weight of zinc gluconate, 0.09 parts by weight of calcium gluconate, 0.09 parts by weight of ferrous gluconate and 1.5 parts by weight of honey in 4 parts by weight of purified water to obtain a calcium, iron and zinc supplement sweetener;

[0071] S4.3: Mix 36 parts by weight of clarifying digestive aid, 27 parts by weight of calcium, iron and zinc supplement sweetener, 5 parts by weight of aromatic digestive aid and 0.15 parts by weight of potassium sorbate evenly, add purified water to make up to 100 parts by weight, then fill into 10 mL vials and sterilize in a 115℃ sterilizer for 30 min to obtain an oral liquid for digestion, calcium, iron and zinc supplementation.

[0072] Example 3

[0073] A method for preparing an oral liquid that aids digestion, relieves indigestion, and supplements calcium, iron, and zinc includes the following steps:

[0074] S1: Extraction and purification of polysaccharides from cauliflower

[0075] S1.1: After cleaning fresh, disease-free cauliflower seedlings, dry them at 50℃ until the moisture content is 8%. Then, crush them and sieve them through a 110-mesh screen to obtain cauliflower powder. Mix the cauliflower powder with distilled water at a mass ratio of 1:40. Boil at 80℃ for 3 hours and then filter. Keep the filtrate and concentrate it under reduced pressure to 25% of the original volume to obtain polysaccharide concentrate.

[0076] S1.2: Add n-butanol-chloroform mixed solvent to the polysaccharide concentrate. The n-butanol-chloroform mixed solvent is prepared by mixing n-butanol and chloroform in a volume ratio of 1:5. The added n-butanol-chloroform mixed solvent accounts for 25% of the volume of the polysaccharide concentrate. Shake thoroughly for 30 min, then centrifuge at 4000 rpm for 15 min. Take the supernatant and repeat the operation until no protein precipitation occurs. Add 4 times the volume of anhydrous ethanol to the last supernatant, let stand overnight at 4℃, centrifuge to collect the precipitate, and dry at 45℃ to obtain cauliflower crude polysaccharide.

[0077] S1.3: Dissolve the crude polysaccharide of *Caulis Amaranthus* in distilled water to prepare a crude polysaccharide solution with a concentration of 6 mg / mL. Then inject the solution into a DEAE cellulose DE-52 column. After eluting the column with distilled water, collect the filtrate, concentrate it to 15% of the original volume, and freeze-dry it at -50℃ to obtain *Caulis Amaranthus* polysaccharide.

[0078] S2: Preparation of digestive aids

[0079] S2.1: Dissolve cauliflower polysaccharide in distilled water at a material-to-liquid ratio of 1:45 g / mL, then add NaOH to adjust the pH of the solution to 9, stir for 15 min, add 5 mL of acetic anhydride dropwise at room temperature while maintaining the pH at 9, stir for 35 min after the acetic anhydride is added, adjust the pH to neutral, and then add it to a dialysis bag with a molecular weight cutoff of 10000 Da for dialysis with deionized water for 3 days. Change the deionized water every 12 h of dialysis. Then take out the solution from the dialysis bag and freeze dry it in a freeze dryer to obtain acetylated cauliflower polysaccharide.

[0080] S2.2: Place hawthorn, coix seed and yam in a container in a mass ratio of 1:1:1, add 30 times the mass of distilled water, decoct at 85℃ for 3 hours, then filter and retain the filtrate, concentrate to a relative density of 1.1 to obtain a mixed concentrate, add 5 wt% acetylated cauliflower polysaccharide to the mixed concentrate, stir until the acetylated cauliflower polysaccharide is completely dissolved to obtain an auxiliary digestive agent.

[0081] S3: Preparation of aromatic digestive agents

[0082] S3.1: Take fresh tender Pu-erh tea leaves and fresh patchouli leaves in a mass ratio of 1:1.2. After washing to remove impurities, dry them in the shade and cut them into strips with a width of 1cm. Then soak them in distilled water for 20 minutes to obtain pre-treated leaves. Place the pre-treated leaves in a volatile oil extraction container and use the steam extraction method. Steam is introduced from the bottom of the extraction container to dry-steam the pre-treated leaves. The temperature in the volatile oil extraction container is controlled at 100℃. The steam passing through the pre-treated leaves is collected through a 25℃ condenser to obtain distilled liquid. Let the distilled liquid stand until it separates into layers and collect the upper oily liquid to obtain Pu-erh-patchouli mixed volatile oil.

[0083] S3.2: Mix 3 parts by weight of food-grade ammonium bicarbonate and 20 parts by weight of hydroxypropyl-β-cyclodextrin evenly, then add 350 parts by weight of distilled water at 25°C, and stir at 350 rpm until clear to obtain an inclusion solution. Under the conditions of 60°C and 650 rpm stirring speed, slowly add 3 parts by weight of Pu-erh tea-patchouli mixed volatile oil to the inclusion solution at the same time for 35 min to obtain a suspension.

[0084] S3.3: Place the suspension in a centrifuge and centrifuge at 5000 rpm for 20 min. Collect the precipitate and dry it under reduced pressure at 70℃ to obtain the aromatic digestive agent.

[0085] S4: Compound oral liquid

[0086] S4.1: Add chitosan to the digestive aid, stir evenly, and allow it to stand and flocculate. The amount of chitosan added is 3wt% of the digestive aid. The flocculation temperature is 35℃ and the flocculation time is 5h. Then filter it with 600-mesh plate and frame filter paper to obtain a clear digestive aid.

[0087] S4.2: Dissolve 0.1 parts by weight of zinc gluconate, 0.1 parts by weight of calcium gluconate, 0.1 parts by weight of ferrous gluconate and 2 parts by weight of honey in 5 parts by weight of purified water to obtain a calcium, iron and zinc supplement sweetener;

[0088] S4.3: Mix 42 parts by weight of clarifying digestive aid, 30 parts by weight of calcium, iron and zinc supplement sweetener, 6 parts by weight of aromatic digestive aid and 0.2 parts by weight of potassium sorbate evenly, add purified water to make up to 100 parts by weight, then fill into 10 mL vials and sterilize in a 115℃ sterilizer for 30 min to obtain an oral liquid for digestion, calcium, iron and zinc supplementation.

[0089] Comparative Example 1

[0090] Steps S1 and S2.1 are removed. In the preparation of the digestive aid in step S2, the acetylated cauliflower polysaccharide in step S2.2 is replaced with an equal mass of hawthorn polysaccharide. The remaining formula and preparation steps are the same as in Example 1.

[0091] Comparative Example 2

[0092] In the preparation of the aromatic digestive agent in step S3, the fresh tender Pu-erh tea leaves in step S3.1 are replaced with an equal mass of fresh patchouli leaves, and the rest of the formula and preparation steps are the same as in Example 1.

[0093] Comparative Example 3

[0094] In the preparation of the aromatic digestive agent in step S3, the fresh patchouli leaves in step S3.1 are replaced with an equal mass of fresh tender Pu-erh tea leaves, and the rest of the formula and preparation steps are the same as in Example 1.

[0095] Comparative Example 4

[0096] In the preparation of the aromatic digestive agent in step S3, the food-grade ammonium bicarbonate in step S3.2 is replaced with an equal mass of hydroxypropyl-β-cyclodextrin, and the remaining formulation and preparation steps are the same as in Example 1.

[0097] Experiment 1: Take the oral liquids prepared in Examples 1-3 and Comparative Example 4, 10 vials in each group, and place them at 40℃ and 75% relative humidity for 3 months. At the end of the first, second and third months, observe whether the oral liquids in each group show turbidity and oil phase separation. Then select one vial for odor identification to judge its aroma. The results are shown in Table 1.

[0098] Table 1: Turbidity and Aroma of Oral Liquid After 3 Months of Storage

[0099]

[0100] As can be seen from the phenomena observed in Examples 1-3 in Table 1 after 3 months of storage, the oral liquid for digestion and supplementing calcium, iron, and zinc prepared in this application has good stability and a fresh woody and herbal spicy aroma. In contrast, Comparative Example 4 did not add food-grade ammonium bicarbonate during the encapsulation of volatile oils, and its aroma was not obvious. This proves that the aromatic digestive agent prepared in this application can prevent volatile oils from seeping into the oral liquid, causing the oral liquid to separate and the taste to deteriorate. At the same time, it can allow the special aromas of Pu-erh tea volatile oil and patchouli volatile oil to be released, thus playing an aromatic therapy role in digestion.

[0101] Experiment 2: Thirty SD rats were selected and randomly divided into three groups of 10 rats each. In addition to normal feeding of the SD rats, one group of SD rats was administered oral solutions prepared in Example 1 and Comparative Example 1 by gavage at a volume of 1 mL / day. The remaining group of rats was set as a blank group and was fed normally every day. After 21 days of feeding, 5 mice were randomly selected from each group and sacrificed. The feces from the cecum of each group were aseptically collected and placed in 2 mL sterile tubes for high-throughput sequencing of intestinal flora. The test results are shown in Table 2.

[0102] Table 2: Significance of abundance of various bacterial species in the intestinal tract of mice after 14 days of feeding

[0103]

[0104] (Mean ± standard deviation. Different letters in the same row indicate significant differences between groups (p < 0.05), and the same letter indicates no significant difference.)

[0105] As can be seen from the data in Table 2 of Example 1 and Comparative Example 1, the oral liquid prepared in Example 1 has a significant inhibitory effect on the proliferation of harmful bacteria such as Proteobacteria, Staphylococcus, Enterococcus, and Bacteroides, while enhancing the proliferation activity of beneficial bacteria such as Firmicutes and Lactobacillus, thereby promoting intestinal digestion and absorption. In contrast, the oral liquid prepared in Comparative Example 1, which replaced acetylated cauliflower polysaccharide with an equal mass of hawthorn polysaccharide, showed lower effects on both the inhibition of harmful bacterial proliferation and the enhancement of beneficial bacterial proliferation activity compared to Example 1. This demonstrates that acetylated cauliflower polysaccharide can inhibit the proliferation of harmful bacteria by promoting the proliferation activity of beneficial bacteria.

[0106] Experiment 3: Ninety male SD rats, weighing between 110-130g, were selected and randomly divided into 9 groups of 10 rats each after 3 days of acclimatization. Each group was administered oral solutions prepared in Examples 1-3 and Comparative Examples 1-4, or distilled water, via gavage. One group of rats received one oral solution, one group of rats received distilled water (as a blank control), and one group of rats received a commercially available digestive oral solution (as a control). The gavage dose was 1 mL / day, administered continuously for 28 days. Rats were kept normally during the experiment, and a clean living environment was maintained.

[0107] Weight test: The initial average weight, average weight at week 4, and weight change of each group of rats were calculated and recorded. The results are shown in Table 3.

[0108] Table 3: Changes in rat body weight

[0109]

[0110] Experiment 4: Ninety Kunming mice were selected, with 10 mice in each group. They were fasted but allowed to drink water. Each group of mice was administered oral solutions prepared in Examples 1-3 and Comparative Examples 1-4, distilled water, and a digestive oral solution by gavage. The distilled water group was set as the blank group, and the commercially available digestive oral solution group was set as the control group. After gavage administration of 0.2 mL of each oral solution for 1 hour, each group was administered 0.2 mL of the oral solution containing 10% carbon powder. After 20 minutes, the mice were euthanized by cervical dislocation. The intestinal segment from the pylorus to the terminal ileum was quickly removed, straightened, and the lengths from the pyloric sphincter to the front of the carbon powder and from the pyloric sphincter to the ileocecal junction were measured.

[0111] The small intestinal propulsion rate was calculated as (length from the pyloric sphincter to the front of the carbon end / length from the pyloric sphincter to the ileocecal junction) × 100%. The average small intestinal propulsion rate of each group of mice was taken, and the results are shown in Table 4.

[0112] Table 4: Small intestinal propulsion rate in mice after drug administration

[0113]

[0114] As can be seen from the data in Tables 3 and 4 of Examples 1-3, the oral liquid for digestion and supplementing calcium, iron and zinc prepared in this application can promote small intestinal motility, thereby effectively enhancing the digestion of food stagnation, and is stronger than the digestive efficacy of commercially available stomach-strengthening and digestive oral liquids.

[0115] Comparative Example 1 shows that without the addition of acetylated cauliflower polysaccharide, digestive function was significantly reduced. This demonstrates that acetylated cauliflower polysaccharide can enhance the digestive function by promoting the proliferation of beneficial bacteria and inhibiting the proliferation of harmful bacteria, thereby promoting the body's absorption and utilization of the effective components of food and effectively improving and treating indigestion.

[0116] The data from Comparative Examples 2 and 3 show that Pu-erh tea volatile oil and patchouli volatile oil have a synergistic effect. The two can work together to provide aromatherapy, thereby promoting digestion and improving indigestion.

[0117] As can be seen from the data of Comparative Example 4, this application mixes ammonium bicarbonate and hydroxypropyl-β-cyclodextrin and encapsulates the Pu-erh-patchouli mixed volatile oil. The tiny channels allow the gas molecules of Pu-erh volatile oil and patchouli volatile oil to dissipate, thereby ensuring the effectiveness of aromatherapy.

[0118] Experiment 5: HepG2 cells and Caco-2 cells were seeded into 96-well plates, respectively, using DEME medium containing 10% fetal bovine serum, 1% non-essential amino acids, and 1% L-glutamine. The cell concentration was adjusted to 1×10⁻⁶ cells / well. 4 / mL, and then incubated in a 37℃ incubator with 5% CO2 for 12h. Then, 2μL of DEME culture medium containing 30% of the oral solution for digestion and calcium, iron and zinc supplementation prepared in Examples 1-3 was added. Negative control wells (without culture medium containing oral solution for digestion and calcium, iron and zinc supplementation) and blank zeroing wells (the final concentration of DMSO in each well was 0.1%) were set up, and the culture was continued for 48h.

[0119] Discard the culture medium from each well, add 20 μL of 5 mg / mL LMT T solution, and continue culturing for 4 h. After discarding the supernatant, dissolve the crystals by shaking with 150 μL LDMSO for 10 min. Measure the optical density (OD) value at 490 nm using an ELISA reader. Use the MTT assay to determine the effect of the easily absorbed oral liquid that promotes the replenishment of both qi and blood on the proliferation of HepG2 and Caco-2 cells. Cell viability (%) = (OD value of the drug well / OD value of the negative control well) × 100%. The results are shown in Table 5.

[0120] Table 5: Cell viability after oral liquid treatment

[0121]

[0122] As can be seen from the data in Examples 1-3 in Table 5, after adding the oral liquid for digestion and supplementing calcium, iron, and zinc prepared in this application to the culture medium for culturing HepG2 cells and Caco-2 cells, the survival rate of HepG2 cells and Caco-2 cells was higher than 96%, which proves that the oral liquid for digestion and supplementing calcium, iron, and zinc prepared in this application has no cytotoxicity and is suitable for human consumption.

[0123] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A method for preparing a calcium, iron and zinc supplement oral liquid for food digestion and accumulation, characterized in that, Comprise the following steps: S1: extraction and purification of polysaccharide from Zostera marina L. Fresh Zostera marina L. seedlings without disease were washed, dried and crushed, then water extraction was carried out, and the obtained polysaccharide concentrate was deproteinized and alcohol precipitated. The obtained Zostera marina L. crude polysaccharide was collected by centrifugation and subjected to DEAE cellulose DE-52 column chromatography to obtain Zostera marina L. polysaccharide; S2: preparation of auxiliary food digestant The Zostera marina L. polysaccharide was dissolved in distilled water, the pH was adjusted to 9, acetic anhydride was added and stirred, then it was adjusted to neutral and dialyzed with deionized water, and acetylated Zostera marina L. polysaccharide was obtained by freeze-drying. The acetylated Zostera marina L. polysaccharide was added to the mixed concentrate obtained by water extraction and boiling of hawthorn, coix seed and yam to obtain an auxiliary food digestant; S3: preparation of aromatic food digestant The fresh chopped Pu'er tea leaves and fresh patchouli leaves were subjected to steam extraction to extract volatile oil to obtain Pu'er-patchouli mixed volatile oil. Food-grade ammonium bicarbonate and hydroxypropyl-β-cyclodextrin were mixed uniformly, then the Pu'er-patchouli mixed volatile oil was slowly added under the conditions of 55-60℃ and 600-650rpm stirring speed. After the addition was completed, centrifugation and reduced pressure drying were carried out to obtain an aromatic food digestant; S4: compounding of oral solution The auxiliary food digestant was flocculated by adding chitosan to obtain a clear auxiliary food digestant. Zinc gluconate, calcium gluconate, ferrous gluconate and honey were dissolved in purified water to obtain a calcium, iron and zinc supplement. The clear auxiliary food digestant, the calcium, iron and zinc supplement, the aromatic food digestant and potassium sorbate were mixed uniformly, then the solution was supplemented, filled, sterilized to obtain a food digestant for treating food accumulation and supplementing calcium, iron and zinc.

2. The method for preparing an oral liquid for digestion, calcium, iron, and zinc supplementation according to claim 1, characterized in that, Step S1 for extraction and purification of polysaccharide from Zostera marina L., specifically comprising the following steps: S1.1: The fresh Zostera marina L. seedlings without disease were washed and dried at 45-50℃, then crushed to pass through a 100-110 mesh sieve to obtain Zostera marina L. powder. The Zostera marina L. powder was mixed with distilled water at a mass ratio of 1: (30-40), then boiled at 75-80℃ for 2.5-3h, filtered, and the filtrate was concentrated under reduced pressure to 20-25% of the original volume to obtain a polysaccharide concentrate; S1.2: The polysaccharide concentrate was added with a mixture of n-butanol-chloroform, shaken thoroughly for 25-30min, then centrifuged at 3000-4000rpm for 10-15min. The supernatant was collected, and the operation was repeated until no protein precipitate was obtained. The obtained supernatant was added with 3-4 times the volume of anhydrous ethanol, and the mixture was placed at 4℃ overnight. The precipitate was collected by centrifugation and dried at 40-45℃ to obtain Zostera marina L. crude polysaccharide; S1.3: The Zostera marina L. crude polysaccharide was dissolved in distilled water to prepare a crude polysaccharide solution with a concentration of 5-6mg / mL, then injected into a DEAE cellulose DE-52 column. The column was eluted with distilled water, and the filtrate was collected, concentrated to 10-15% of the original volume, and freeze-dried at -40 to -50℃ to obtain Zostera marina L. polysaccharide.

3. The method for preparing an oral liquid for digestion, supplementing calcium, iron, and zinc according to claim 2, characterized in that, Step S2 for preparation of auxiliary food digestant, specifically comprising the following steps: S2.1: dissolve the polysaccharide of Zostera marina L. in distilled water at a solid-liquid ratio of 1: (40-45) g / mL, then add NaOH to adjust the pH of the solution to 9, stir for 10-15 min, add 3-5 mL of acetic anhydride drop by drop at room temperature and maintain the pH at 9, stir for 30-35 min after the addition of acetic anhydride is completed, adjust the pH to neutral, then put the solution in a dialysis bag and dialyze with deionized water, then take out the solution in the dialysis bag and freeze-dry in a freeze dryer to obtain acetylated Zostera marina L. polysaccharide; S2.2: put hawthorn, coix seed and yam in a container at a mass ratio of 1: (0.5-1): (0.5-1), add 25-30 times the mass of distilled water, then cook at 80-85℃ for 2-3h, then filter to retain the filtrate, concentrate to a relative density of 1.08-1.1 to obtain a mixed concentrate, add 4-5wt% acetylated Zostera marina L. polysaccharide to the mixed concentrate, stir until the acetylated Zostera marina L. polysaccharide is completely dissolved to obtain an auxiliary food digestant.

4. The method for preparing an oral liquid for digestion, calcium, iron, and zinc supplementation according to claim 3, characterized in that, Step S3 for preparing the aromatic food digestant, specifically comprising the following steps: S3.1: take fresh tender Pu'er tea leaves and fresh plectranthus amboinicus leaves at a mass ratio of 1: (1-1.2), wash with water to remove impurities, then dry in the shade and cut into strips with a width of 0.5-1cm, then soak in distilled water for 10-20min to obtain pretreated leaves, put the pretreated leaves in a volatile oil extraction container, use steam extraction method to dry the pretreated leaves by passing steam from the bottom of the extraction container, control the temperature in the volatile oil extraction container at 95-100℃, and collect the steam passing through the pretreated leaves by a 20-25℃ condensing device to obtain a distillate, let the distillate stand to separate into layers, collect the upper oily liquid to obtain a Pu'er-plectranthus amboinicus mixed volatile oil; S3.2: mix 2-3 parts by weight of food-grade ammonium bicarbonate and 18-20 parts by weight of hydroxypropyl-β-cyclodextrin uniformly, then add 300-350 parts by weight of 20-25℃ distilled water, stir at a speed of 300-350rpm until clear to obtain an inclusion solution, slowly add 2-3 parts by weight of Pu'er-plectranthus amboinicus mixed volatile oil to the inclusion solution at 55-60℃ and a stirring speed of 600-650rpm, and the addition time is 30-35min to obtain a suspension; S3.3: centrifuge the suspension in a centrifuge at 4000-5000rpm for 15-20min, collect the precipitate and dry at 65-70℃ under reduced pressure to obtain an aromatic food digestant.

5. The preparation method of an oral liquid for digestion, calcium, iron, and zinc supplementation according to claim 4, characterized in that, Step S4 for compounding the oral liquid, specifically comprising the following steps: S4.1: add chitosan to the auxiliary food digestant and stir uniformly, let it stand and flocculate, then filter with a plate and frame filter 600 mesh filter paper to obtain a clear auxiliary food digestant; S4.2: dissolve 0.08-0.1 parts by weight of zinc gluconate, 0.08-0.1 parts by weight of calcium gluconate, 0.08-0.1 parts by weight of ferrous gluconate and 1-2 parts by weight of honey in 3-5 parts by weight of purified water to obtain a calcium, iron and zinc supplement sweetener; S4.3: 30-42 parts by weight of the clarified auxiliary digestion agent, 24-30 parts by weight of the calcium, iron and zinc supplement, 4-6 parts by weight of the aromatic digestion agent and 0.08-0.1 parts by weight of potassium sorbate are mixed uniformly, purified water is added to make up to 100 parts by weight, then each 10 mL is filled, and placed in a 115°C sterilization cabinet for 30 min sterilization to obtain the oral liquid of digestion and accumulation of calcium, iron and zinc supplement.

6. The method for preparing an oral liquid for digestion, relieving indigestion, and supplementing calcium, iron, and zinc according to claim 2, characterized in that, The n-butanol-chloroform mixed solvent in step S1.2 is prepared by mixing n-butanol and chloroform at a volume ratio of 1:5, and the n-butanol-chloroform mixed solvent added accounts for 20-25% of the volume of the polysaccharide concentrated solution.

7. The method for preparing an oral liquid for digestion, calcium, iron, and zinc supplementation according to claim 3, characterized in that, The molecular weight cut-off of the dialysis bag in step S2.1 is 8000-10000 Da.

8. The method for preparing an oral liquid for digestion, relieving indigestion, and supplementing calcium, iron, and zinc according to claim 3, characterized in that, The dialysis time in step S2.1 is 2-3 days, and the deionized water is replaced every 8-12 hours.

9. The method for preparing an oral liquid for digestion, calcium, iron, and zinc supplementation according to claim 5, characterized in that, The addition amount of chitosan in step S4.1 accounts for 2-3 wt% of the auxiliary digestion agent, the flocculation temperature is 30-35°C, and the flocculation time is 4-5 h.

10. A kind of oral liquid of digestion and accumulation of eliminating food, characterized by, It is prepared by the preparation method of the oral liquid of digestion and accumulation of calcium, iron and zinc supplement of any one of claims 1-9.

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