Sargassum pallidum polysaccharide oral liquid as well as preparation method and quality evaluation thereof

Abstract

The invention discloses sargassum pallidum polysaccharide oral liquid as well as a preparation method and quality evaluation thereof, and belongs to the technical field of pharmaceutical preparations. The oral liquid is prepared by taking sargassum pallidum polysaccharide as a main medicine and matching sucralose, potassium sorbate, chitosan and other auxiliary materials through pH regulation, chitosan flocculation and centrifugal purification processes. Chitosan is directly added after the pH value of the solution is adjusted to be acidic, and meanwhile, the taste is remarkably improved by screening and optimizing a flavoring agent. The process is efficient, the cost is controllable, and the obtained oral liquid is high in clarity, excellent in active ingredient retention and suitable for industrial production.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical preparations, and in particular to an oral solution of Artemisia seagrass polysaccharide and a preparation method and quality evaluation thereof. Background Art

[0002] With the rapid development of life science and medical technology, marine biological resources have become a hot area for scientific research and industrial development due to their rich bioactive substances. As a typical representative of marine algae, the polysaccharides contained in sea wormwood have great application value in the pharmaceutical and health care industries due to their unique biological activity, and have attracted widespread attention.

[0003] Artemisia sea polysaccharides have significant antioxidant properties, can effectively capture and remove excess free radicals in the body, alleviate the damage caused by oxidative stress to cells and tissues, and provide a new way to prevent and treat oxidative stress-related diseases such as cardiovascular disease and neurodegenerative diseases. In terms of immune regulation, Artemisia sea polysaccharides can stimulate the activation and proliferation of immune cells, enhance the body's immune defense capabilities, and enhance the body's resistance to various pathogens. It has potential application prospects in the prevention and adjuvant treatment of infectious diseases. At the same time, its anti-tumor properties have also been confirmed by many studies. By inducing tumor cell apoptosis and hindering the growth and metastasis of tumor cells, it has opened up a new direction for the research and development of tumor treatment drugs. In addition, Artemisia sea polysaccharides also show a positive effect in regulating blood lipid metabolism and lowering blood lipid levels, which is of great significance for the prevention and treatment of metabolic diseases such as hyperlipidemia.

[0004] However, the current process of sea wormwood polysaccharide from laboratory research to industrial application is faced with a series of key problems that need to be solved. In the extraction process, the traditional extraction process generally has the disadvantages of low polysaccharide extraction rate and poor purity, which not only causes the waste of precious marine resources, but also greatly increases the production cost, limiting the large-scale production and market promotion of sea wormwood polysaccharide products. In the preparation stage, how to ensure the stability and biological activity of sea wormwood polysaccharide in a complex preparation environment has become a bottleneck restricting product quality and efficacy. Factors such as pH, temperature changes and ionic strength fluctuations in the solution system may cause structural changes or degradation of sea wormwood polysaccharide, thereby reducing its biological activity. At the same time, the sea wormwood polysaccharide extract is often mixed with proteins, pigments, small molecule impurities, etc., causing the extract to be turbid, seriously affecting the clarity of the preparation, not only reducing the appearance quality of the product, but also may have a negative impact on the stability and shelf life of the product.

[0005] In terms of taste optimization, the sour and astringent taste of sea wormwood polysaccharide itself greatly reduces the palatability of the preparation, seriously affecting consumers' acceptance and compliance. The current flavoring agents on the market have different effects in covering up the unpleasant taste of sea wormwood polysaccharide. Although some flavoring agents can effectively cover up the sour and astringent taste, they often bring new problems of being too sweet and greasy, affecting the overall taste experience of the product; some flavoring agents may interact with sea wormwood polysaccharide, posing a potential threat to its stability and biological activity.

[0006] In summary, the development of a set of efficient, stable preparation processes for Artemisia marinum polysaccharide preparations that take both taste and quality into consideration has great practical significance and urgent market demand. The present invention is based on a profound understanding and in-depth research on the above-mentioned problems, and aims to provide a practical solution for the development of Artemisia marinum polysaccharide preparations through innovative technical means and scientific experimental design, and promote the industrialization process of Artemisia marinum polysaccharide-related products. Summary of the invention

[0007] The technical problem to be solved by the present invention is to provide an oral solution of Artemisia seaweed polysaccharide and a preparation method and quality evaluation thereof in view of the deficiencies in the prior art.

[0008] In order to achieve the purpose of the present invention, the following technical means are specifically adopted:

[0009] An oral solution of Artemisia seaweed polysaccharide, characterized in that it is composed of the following ingredients:

[0010] Main drug: Artemisia seaweed polysaccharide 0.1g / ml preparation;

[0011] Flavoring agent: sucralose 0.002-0.006g / ml preparation;

[0012] Preservative: Potassium sorbate 0.001 g / ml preparation;

[0013] Clarifier: chitosan 0.0008-0.0032g / ml preparation;

[0014] pH adjusters: sodium dihydrogen phosphate and citric acid;

[0015] Solvent: Make up to the required volume with purified water.

[0016] The artemisia seaweed polysaccharide oral liquid is characterized in that the added amount of chitosan is 0.0032 g / ml preparation.

[0017] The said Artemisia seagrass polysaccharide oral liquid is characterized in that the added amount of the flavor corrective sucralose is 0.004 g / ml preparation.

[0018] The said Artemisia seaweed polysaccharide oral liquid is characterized in that the clarifier chitosan is added in a manner of directly adding chitosan powder after adjusting the solution pH to 5.0.

[0019] The method for preparing the artemisia seaweed polysaccharide oral liquid is characterized by comprising the following steps:

[0020] (1) Weigh the prescribed amount of Artemisia seaweed polysaccharide powder and add 90% of the prescribed amount of purified water to dissolve;

[0021] (2) adjusting the solution pH to 4.0-6.0 with sodium dihydrogen phosphate and citric acid;

[0022] (3) Add chitosan and stir in a constant temperature water bath at 40-80°C for 30 minutes;

[0023] (4) Centrifuge at 4000 r / min for 10 min and filter the supernatant;

[0024] (5) Add sucralose as a flavoring agent and potassium sorbate as a preservative, and make up the volume with purified water.

[0025] The preparation method is characterized in that the temperature of the constant temperature water bath in step (3) is 60°C.

[0026] The preparation method is characterized in that in step (2), the pH is adjusted to 5.0.

[0027] The sea wormwood polysaccharide oral liquid is characterized in that the raw material quality standards of the sea wormwood polysaccharide are: polysaccharide content ≥50% (in terms of glucose), drying loss ≤1%, ignition residue ≤0.2%, heavy metal ≤10ppm, pesticide residue ≤2ppm, and microbial limit meets the requirements of the pharmacopoeia.

[0028] The application of the sea wormwood polysaccharide oral liquid in the preparation of medicines or health products with antioxidant, immunomodulatory or anti-tumor functions.

[0029] The sea wormwood polysaccharide described in the present invention is powdery and brown, with a polysaccharide content (calculated as glucose) of ≥50%, a drying loss of ≤1%, an ignition residue of ≤0.2%, heavy metals of ≤10ppm, pesticide residues of ≤2ppm, a total bacterial count of <1000CFU / g, molds and yeasts of <100CFU / g, no salmonella detected, and no Escherichia coli detected.

[0030] Beneficial Effects

[0031] Beneficial effects of the present invention:

[0032] 1. In response to the sour and astringent taste of Artemisia seaweed polysaccharide, through the screening of flavoring agents and the addition amount gradient experiment, it was determined that 0.4% sucralose can effectively mask the sour and astringent feeling and has moderate sweetness.

[0033] 2. The present invention achieves an ultra-high polysaccharide retention rate under optimal process parameters by systematically optimizing the synergistic effect of chitosan addition amount, flocculation temperature and system pH value. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 Glucose standard curve. DETAILED DESCRIPTION

[0035] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] Description of the sea wormwood polysaccharide used in the present invention: the manufacturer is Shaanxi Mixianer Biotechnology Co., Ltd., the product batch number is: 2024.08.21, the polysaccharide content (calculated as glucose) is 50.1%, the drying loss is 0.54%, the ignition residue is 0.12%, the heavy metal is ≤10ppm, the pesticide residue is ≤2ppm, the total bacteria count is <1000CFU / g, the mold and yeast are <100CFU / g, Salmonella shall not be detected, and Escherichia coli shall not be detected.

[0037] Example 1

[0038] 1. The preliminary prescription is as follows:

[0039] Element effect Material addition amount / ml preparation Artemisia seaweed polysaccharide Main drug 0.1g Xylitol Flavoring agents 0.006g Potassium Sorbate preservative 0.001g Chitosan Clarifier 0.0032g Sodium dihydrogen phosphate pH Adjusters Moderate Citric acid pH Adjusters Moderate Purified water Solvent Make up to 1ml

[0040] 2. The preliminary preparation method is as follows:

[0041] Weigh the prescribed amount of Artemisia seagrass polysaccharide powder, add 90% of the prescribed amount of purified water to dissolve it, adjust the pH to 5.0 with sodium dihydrogen phosphate and citric acid, add chitosan as a clarifier, keep in a constant temperature water bath at 60°C for 30 minutes, centrifuge at 4000r / min for 10 minutes, filter, take the supernatant, add flavoring agents and preservatives, add purified water to make up to volume, and obtain the product.

[0042] 3. The calculation and measurement methods of some evaluation indicators are as follows:

[0043] 3.1 Polysaccharide retention rate

[0044] Polysaccharide retention rate (%) = polysaccharide content in the sample after treatment / polysaccharide content in the sample before treatment × 100%

[0045] 3.2 Determination of polysaccharide content in preparations

[0046] The method for determining the content of the polysaccharide in the present invention is to adopt the sulfuric acid-anthrone method.

[0047] A. Preparation of standard solution:

[0048] (1) Anthrone reagent: Accurately weigh 0.1 g of anthrone, add 80% concentrated sulfuric acid to dissolve it, transfer to a brown volumetric flask, dilute to 100 ml with 80% concentrated sulfuric acid, and shake well; use immediately after preparation (not more than 2 hours).

[0049] (2) Glucose standard solution: Place glucose in a phosphorus pentoxide desiccator. After 12 hours, accurately weigh 0.1 g of glucose and dilute to 100 ml with distilled water to prepare a 1.0 mg / ml glucose standard solution.

[0050] B. Drawing of standard curve: Accurately pipette 0ml, 0.10ml, 0.20ml, 0.30ml, 0.40ml, 0.50ml and 0.60ml of glucose standard solution into 10ml stoppered test tubes, and make up to 1.0ml with distilled water. Add 4.0ml of anthrone-sulfuric acid solution respectively, use a vortex oscillator to mix the reaction solution, and then place the test tube in a boiling water bath to react for 10 minutes. Cool to room temperature in a cold water bath for 10 minutes, and immediately measure the absorbance at 626nm using a UV-visible spectrophotometer. Use the mass of glucose as the horizontal axis and the absorbance as the vertical axis to draw up a standard curve, see Figure 1 .

[0051] C. Determination of sample content: Take 1.0 ml of sample solution, dilute it by an appropriate multiple, and measure the absorbance of the polysaccharide sample at 626 nm according to the above method to calculate the polysaccharide content.

[0052] Example 2

[0053] 1. Effect of chitosan addition on polysaccharide retention rate

[0054] According to the preliminary prescription, the polysaccharide powder of Artemisia seagrass was weighed, and 90% of the prescribed amount of purified water was added to dissolve it, and the pH was adjusted to 5.0 with sodium dihydrogen phosphate and citric acid. Six portions of the above solution were prepared in parallel, and 0.08%, 0.16%, 0.24%, 0.32%, 0.40% and 0.48% of chitosan were added respectively (i.e., 0.08, 0.16, 0.24, 0.32, 0.40 and 0.48 g of chitosan were added to every 100 ml of the final preparation). After flocculation at 60°C for 30 minutes, the solution was placed in a centrifuge and centrifuged at 4000 r / min for 10 minutes. The solution was allowed to stand and filtered. The supernatant was taken, flavoring agents and preservatives were added, and the volume was fixed with purified water. The absorbance was determined, and the polysaccharide retention rate was calculated.

[0055] Chitosan addition amount (%) Polysaccharide retention rate (%) 0.08 96.58 0.16 96.76 0.24 97.92 0.32 99.29 0.40 84.01 0.48 83.44

[0056] According to the above research results, the chitosan dosage is in the range of 0.08-0.32% (w / v), and the polysaccharide retention rate is high (>96.58%), which is within the acceptable range. Among them, the polysaccharide retention rate is the highest when the chitosan dosage is 0.32% (w / v), and the chitosan dosage of 0.32% (w / v) is temporarily selected for subsequent investigation and research.

[0057] 2. Effect of flocculation temperature on polysaccharide retention rate

[0058] According to the preliminary prescription, weigh the sea wormwood polysaccharide powder, add 90% of the prescription amount of purified water to dissolve it, adjust the pH to 5.0 with sodium dihydrogen phosphate and citric acid, add 0.32% chitosan, prepare 6 portions of the above solution in parallel, stir in a water bath at flocculation temperatures of 30°C, 40°C, 50°C, 60°C, 70°C and 80°C, respectively, keep warm and flocculate for 30 minutes, put into a centrifuge, centrifuge at 4000r / min for 10 minutes, let it stand, filter, take the supernatant, add flavoring agents and preservatives, make up to volume with purified water, measure its absorbance, and calculate its polysaccharide retention rate.

[0059] Flocculation temperature (℃) Polysaccharide retention rate (%) 30 88.94 40 94.05 50 94.90 60 99.78 70 98.67 80 96.32

[0060] According to the above research results, the retention rate of polysaccharides is relatively high (>94.05%) when the flocculation temperature is between 40 and 80°C, which is within the acceptable range. Among them, the polysaccharide retention rate is the highest when the flocculation temperature is 60°C, so the flocculation temperature of 60°C is temporarily selected for subsequent investigation and research.

[0061] 3. Effect of system pH on polysaccharide retention rate

[0062] According to the preliminary prescription, weigh the prescribed amount of Artemisia seagrass polysaccharide powder, add 90% of the prescribed amount of purified water to dissolve it, prepare 5 portions of the above solution in parallel, adjust the pH to 4.0, 5.0, 6.0, 7.0 and 8.0 respectively with sodium dihydrogen phosphate and citric acid, add 0.32% chitosan, stir in a water bath at 60°C flocculation temperature, keep warm and flocculate for 30 minutes, put into a centrifuge, centrifuge at 4000r / min for 10 minutes, let stand, filter, take the supernatant, add flavoring agents and preservatives, make up to volume with purified water, measure its absorbance, and calculate its polysaccharide retention rate.

[0063] System pH Polysaccharide retention rate (%) 4.0 95.70 5.0 99.56 6.0 98.49 7.0 85.57 8.0 85.35

[0064] According to the above research results, the retention rate of polysaccharides is relatively high (>95.70%) when the system pH value is between 4.0 and 6.0, which is within the acceptable range. Among them, the polysaccharide retention rate is the highest when the system pH value is 5.0, so the system pH value of 5.0 is temporarily selected for subsequent investigation and research.

[0065] 4. Screening of flavoring agents

[0066] According to the optimal experimental conditions investigated in the above experiments, an oral solution of Artemisia seagrass polysaccharide was prepared, and the amount of flavoring agent added was 0.6% (w / v). The taste of the oral solution when the flavoring agent was sucralose, xylitol, and stevioside was compared.

[0067] Flavoring agents Taste Sucralose Too sweet, but can cover up the sourness of the original liquid Xylitol Slightly sweet, but can't cover up the sourness Stevioside Sour, slightly bitter

[0068] Based on the above investigation results, sucralose was selected as the flavor corrector for subsequent research.

[0069] 5. Screening of the amount of flavoring agent added

[0070] According to the optimal experimental conditions investigated in the above experiments, an oral solution of Artemisia seaweed polysaccharide was prepared, and sucralose was selected as the flavoring agent. The taste of the oral solution was compared when the flavoring agent was added in an amount of 0.2%, 0.4%, 0.6%, 0.8% and 1.0% (w / v).

[0071] Flavoring agent added (%) Taste 0.2 Can mask the bitter taste of the drug, but has a sour and astringent taste 0.4 Can mask the bitter taste of the drug, and the overall sweetness is moderate 0.6 It can mask the bitter taste of the drug, but the taste is too sweet 0.8 Too sweet taste 1.0 Too sweet taste

[0072] According to the investigation results, the dosage of flavoring agent added in the prescription was selected as 0.4% (w / v) for subsequent research.

[0073] 6. Screening of chitosan addition methods

[0074] Chitosan is a water-insoluble substance that can be dissolved in acidic solutions. There are two common methods for adding chitosan to a solution: (1) making a chitosan-acetic acid solution and adding it to the drug solution; (2) adjusting the pH value of the solution to acidic before adding chitosan. The two methods were tested separately and compared in terms of flocculation state, operation status, and polysaccharide retention rate to select the best addition method.

[0075] Method 1: Chitosan-acetic acid solution method

[0076] Prepare chitosan solution with a mass fraction of 6.4% (g / ml) with 1% acetic acid solution. Weigh the prescribed amount of Artemisia seagrass polysaccharide powder, add 90% of the prescribed amount of purified water to dissolve, measure chitosan-acetic acid solution, add chitosan to keep the proportion of chitosan in the prescribed amount at 0.32% (w / v), keep it in a 60°C constant temperature water bath for 30 minutes, centrifuge at 4000r / min for 10 minutes, filter, take the supernatant, add 0.4% (w / v) sucralose as a flavoring agent and potassium sorbate as a preservative, add purified water to make up to volume, and obtain the solution.

[0077] Method 2: Adjusting the pH value of the solution

[0078] Weigh the prescribed amount of Artemisia seagrass polysaccharide powder, add 90% of the prescribed amount of purified water to dissolve it, adjust the pH to 5.0 with sodium dihydrogen phosphate and citric acid, add 0.32% chitosan, stir in a water bath at 60°C flocculation temperature, keep warm and flocculate for 30 minutes, put it into a centrifuge, centrifuge at 4000r / min for 10 minutes, let it stand, filter, take the supernatant, add flavoring agents and preservatives, and make up to volume with purified water to obtain the product.

[0079] Chitosan Addition Method Polysaccharide retention rate (%) Flocculation status Operation Chitosan-acetic acid solution method 98.80 Visually good Chitosan takes a long time to dissolve and is time-consuming Solution pH adjustment method 100.09 Visually good Easy to operate

[0080] According to the above investigation results, chitosan was added by adjusting the solution pH value.

[0081] Example 3

[0082] Samples were prepared according to the prescription and preparation method screened out in the above experiment for quality evaluation.

[0083] Element effect Material addition amount / ml preparation Artemisia seaweed polysaccharide Main drug 0.1g Sucralose Flavoring agents 0.004g Potassium Sorbate preservative 0.001g Chitosan Clarifier 0.0008～0.0032g Sodium dihydrogen phosphate pH Adjusters Moderate Citric acid pH Adjusters Moderate Purified water Solvent Make up to 1ml

[0084] Note: The content of 1 ml written here is only used to clearly indicate the proportion of each component in the prescription, and does not mean that the unit dose of the Artemisia seagrass polysaccharide oral solution described in the present invention is 1 ml.

[0085] Weigh the prescribed amount of Artemisia seagrass polysaccharide powder, add 90% of the prescribed amount of purified water to dissolve it, adjust the pH to 4.0-6.0 with sodium dihydrogen phosphate and citric acid, add chitosan as a clarifier, keep in a constant temperature water bath at 40-80°C for 30 minutes, centrifuge at 4000r / min for 10 minutes, filter, take the supernatant, add flavoring agents and preservatives, add purified water to make up to volume, and obtain the product.

[0086] 1. Traits

[0087] This product is a brown clear liquid with a slightly sweet taste.

[0088] 2. pH value

[0089] The pH value of this product was measured to be 5.20.

[0090] 3. Relative density

[0091] Take 3 batches of samples, determine the relative density according to the method under the 2020 edition of the "Chinese Pharmacopoeia", and use the pycnometer method to measure the relative density of the oral liquid. Accurately weigh the dry and clean pycnometer, then fill the pycnometer with oral liquid and place it in a water bath to make the temperature of the oral liquid reach 20°C, then take out the pycnometer, wipe the bottle dry, accurately weigh the weight, subtract the weight of the pycnometer itself, and get the weight of the oral liquid. Use the same method to get the weight of freshly boiled cold water. Calculate the relative density of the oral liquid according to the formula. Relative density of oral liquid = weight of oral liquid / weight of water.

[0092] The relative density values ​​of the three oral solutions were 1.016, 1.017, and 1.016 g / ml, respectively, with an average value of 1.016 g / ml.

[0093] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An oral solution of Artemisia seaweed polysaccharide, characterized in that: It is composed of the following ingredients: Main drug: Artemisia seaweed polysaccharide 0.1g / ml preparation; Flavoring agent: sucralose 0.002-0.006g / ml preparation; Preservative: Potassium sorbate 0.001 g / ml preparation; Clarifier: chitosan 0.0008-0.0032g / ml preparation; pH adjusters: sodium dihydrogen phosphate and citric acid; Solvent: Make up to the required volume with purified water.

2. The oral solution of Artemisia seagrass polysaccharide according to claim 1, characterized in that: The added amount of chitosan is 0.0032 g / ml preparation.

3. The oral solution of Artemisia seagrass polysaccharide according to claim 1, characterized in that: The flavor correcting agent sucralose is added in an amount of 0.004 g / ml of the preparation.

4. The oral solution of Artemisia seagrass polysaccharide according to claim 1, characterized in that: The method of adding the clarifier chitosan is: directly adding chitosan powder after adjusting the solution pH to 5.

0.

5. A method for preparing the oral solution of Artemisia seagrass polysaccharide according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) Weigh the prescribed amount of Artemisia seaweed polysaccharide powder and add 90% of the prescribed amount of purified water to dissolve; (2) adjusting the pH of the solution to 4.0-6.0 with sodium dihydrogen phosphate and citric acid; (3) Add chitosan and stir in a constant temperature water bath at 40-80°C for 30 minutes; (4) Centrifuge at 4000 r / min for 10 min and filter the supernatant; (5) Add sucralose as a flavor enhancer and potassium sorbate as a preservative, and make up the volume with purified water.

6. The preparation method according to claim 5, characterized in that: The temperature of the constant temperature water bath in step (3) is 60°C.

7. The preparation method according to claim 5, characterized in that: In step (2), the pH is adjusted to 5.

0.

8. The oral solution of Artemisia seagrass polysaccharide according to claim 1, characterized in that: The raw material quality standards of the sea wormwood polysaccharide are: polysaccharide content ≥50% (in terms of glucose), drying loss ≤1%, ignition residue ≤0.2%, heavy metal ≤10ppm, pesticide residue ≤2ppm, and microbial limit in accordance with the pharmacopoeia.

9. Use of the oral solution of Artemisia seagrass polysaccharide according to any one of claims 1 to 8 in the preparation of medicines or health products with antioxidant, immunomodulatory or anti-tumor functions.

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