A method for preparing an anti-fatigue chewing gum

Abstract

This invention discloses a method for preparing anti-fatigue chewing gum, comprising the following steps: weighing the following components according to the specified ratio: Yunnan olive extract, white sugar, maltodextrin, purslane extract, ginseng leaf extract, glucose powder, gelatin, coating colorant, and steviol glycosides; mixing them evenly; adding water; heating and stirring until completely dissolved; molding the mixture; and coating it to obtain the anti-fatigue chewing gum. The anti-fatigue chewing gum provided by this invention uses extracts of medicinal and edible herbs as its main functional ingredients, exhibits significant anti-fatigue effects, and has a pleasant taste, is easy to carry, and can be swallowed directly.

Description

Technical Field

[0001] This invention belongs to the field of chewing gum preparation technology, specifically, it relates to a method for preparing anti-fatigue chewing gum. Background Technology

[0002] Fatigue is a common physiological response under prolonged high-intensity work conditions, manifesting as poor concentration, slow reaction time, and decreased work efficiency, severely impacting work efficiency and safety, and even potentially leading to work accidents. Currently, methods to alleviate fatigue mainly include energy supplements and neurostimulants, such as L-carnitine, creatine monohydrate, coenzyme Q10, caffeine, amphetamine, and modafinil. However, products such as Rhodiola rosea capsules, compound amino acid capsules, high-oxygen solutions, and green tea active extracts suffer from problems such as inconvenience in carrying, limited usage scenarios, and high costs, making them difficult to promote and apply in special working environments.

[0003] Traditional Chinese medicine (TCM) possesses unique advantages in combating fatigue. Guided by the theory of "combining tonification and purgation," it can both replenish vital energy and eliminate pathogenic factors, effectively addressing the core pathogenesis of fatigue caused by deficiency of vital energy and stagnation of pathogenic factors. Purslane has the effects of clearing heat and detoxifying, cooling blood and stopping bleeding; modern research shows that it has anti-hypoxia and anti-fatigue effects. Ginseng leaves have the effects of tonifying qi and benefiting the lungs, relieving summer heat and promoting body fluid production; ginsenosides are its main active ingredient for combating fatigue. Combining the two can exert a synergistic anti-fatigue effect. However, both purslane and ginseng leaf extracts have a strong bitter taste, and conventional extracts have high yields and many impurities, making them difficult to use directly in chewing gum preparations. Therefore, developing a palatable, portable, safe and effective anti-fatigue chewing gum is of significant practical importance. Summary of the Invention

[0004] The purpose of this invention is to provide a method for preparing anti-fatigue chewing gum.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] In a first aspect, the present invention provides a method for preparing anti-fatigue chewing gum, comprising the following steps:

[0007] The anti-fatigue chewing gum is made from the following components by weight percentage: 40-50% Yunnan olive extract, 20-25% white sugar, 8-16% maltodextrin, 5-10% purslane extract, 2-8% ginseng leaf extract, 1-5% glucose powder, 0.1-2% gelatin, 0.01-1% coating colorant, and 0.005-0.1% steviol glycosides.

[0008] Weigh out the following components according to the specified ratio: Yunnan olive extract, white sugar, maltodextrin, purslane extract, ginseng leaf extract, glucose powder, gelatin, coating colorant, and steviol glycosides. Mix them evenly, add water, heat and stir until completely dissolved, shape the mixture, and coat it to obtain the anti-fatigue chewing gum.

[0009] Preferably, the anti-fatigue chewing gum is made from the following components in weight percentage: 47.4% Yunnan olive extract, 22.48% white sugar, 12% maltodextrin, 8.3% purslane extract, 5.6% ginseng leaf extract, 3% glucose powder, 1% gelatin, 0.2% coating colorant, and 0.02% steviol glycosides.

[0010] Preferably, the anti-fatigue chewing gum is made from the following components in weight percentage: 45.6% Yunnan olive extract, 21.3% white sugar, 13.2% maltodextrin, 10.88% purslane extract, 5% ginseng leaf extract, 2.8% glucose powder, 1% gelatin, 0.2% coating colorant, and 0.02% steviol glycosides.

[0011] The preparation method of the ginseng leaf extract includes the following steps:

[0012] Add 8-14 times (preferably 12 times) of 40-95% (preferably 50%) ethanol to the coarse ginseng leaf segments and decoct 1-3 times (preferably 2 times) for 0.5-2 hours (preferably 2 hours). Filter, combine the filtrates, concentrate under reduced pressure, add water, filter through a microporous membrane, dilute, and purify to obtain the ginseng leaf extract.

[0013] Preferably, the preparation method of the ginseng leaf extract includes the following steps:

[0014] The coarse ginseng leaf segments were decocted twice with 12 times the amount of 50% ethanol for 2 hours each time. The filtrates were then combined, concentrated under reduced pressure, and water was added. The mixture was then filtered through a microporous membrane, diluted, and purified to obtain the ginseng leaf extract.

[0015] The purification was performed using a D101 macroporous resin column, eluting sequentially with water, 40% ethanol, and 95% ethanol, and collecting the 40% ethanol eluent.

[0016] The preparation method of the purslane extract includes the following steps: purslane slices are extracted by reflux with 8-20 times (preferably 15 times) of 50-95% ethanol (preferably 80% ethanol) (extracted 1-3 times, preferably once; extraction time 0.5-1.5 h, preferably 0.5 h), concentrated under reduced pressure, dissolved in distilled water, and the pH is adjusted to 6.5-7.0 with 5-15% (preferably 10%) NaOH solution. The mixture is then centrifuged (preferably at 5000 rpm for 1 min), the precipitate is washed with water (preferably once), and vacuum dried (at a temperature of 50-80℃, preferably 60℃, for 6-22 h, preferably 8 h) to obtain the purslane extract.

[0017] Preferably, the preparation method of the purslane extract includes the following steps: purslane slices are extracted once by reflux with 15 times 80% ethanol for 0.5 h; the extraction is concentrated under reduced pressure, dissolved in distilled water, and the pH is adjusted to 6.5-7.0 with 10% NaOH solution. The mixture is then centrifuged at 5000 rpm for 1 min, the precipitate is washed once with water, and then vacuum dried at 60℃ for 8 h to obtain the purslane extract.

[0018] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:

[0019] The anti-fatigue chewing gum provided by this invention uses extracts of Chinese medicinal herbs that are both food and medicine as its main functional ingredients. It has a significant anti-fatigue effect, and also has a good taste, is easy to carry, and can be swallowed directly.

[0020] This invention optimizes the preparation processes of purslane extract and ginseng leaf extract, determining the optimal extraction, purification, and drying conditions. These extracts are then combined with excipients such as Yunnan olive extract. Through taste adjustment and chewing time optimization, the resulting chewing gum has a pleasant taste, can be swallowed directly, leaves no residue, is easy to carry, and exhibits significant anti-fatigue effects. Pharmacological and preliminary clinical studies have shown that this product can significantly reduce scores on mental fatigue scales and is suitable for people engaged in high-intensity mental work.

[0021] This invention provides an anti-fatigue chewing gum, the first of its kind to carry traditional Chinese medicine extracts in chewing gum form to relieve mental fatigue and reduce the risk of training injuries. The product is cost-effective, has a simple preparation process, and is guided by the traditional Chinese medicine theory of "combining tonification and purgation" in anti-fatigue formulations, addressing the core pathogenesis of fatigue caused by deficiency of vital energy and stagnation of pathogenic factors. This formulation theory significantly differs from the currently prevalent theories of "pure tonification without purgation" or "receptor depletion" in anti-fatigue drug formulations, demonstrating its advanced nature.

[0022] The anti-fatigue chewing gum provided by this invention combines the readily accepted functional food ingredients, purslane and ginseng leaves (both medicinal and edible), into a chewing gum formulation that is easy to carry and consume. This represents a new approach to the development of anti-fatigue functional nutritional supplements. It improves upon the situation where workers in related occupations can only rely on passive recovery methods such as sleep to alleviate mental fatigue, providing a new intervention approach and means to reduce the risks associated with mental fatigue. Attached Figure Description

[0023] Figure 1 This is a schematic diagram showing the relationship between sample solution concentration and resin adsorption capacity.

[0024] Figure 2 This is a schematic diagram of the adsorption curve results of ginsenoside Re from ginseng leaves on D101 type resin.

[0025] Figure 3 This is a schematic diagram showing the relationship between flow rate and sample adsorption capacity.

[0026] Figure 4 This is a schematic diagram of the desorption curve results of dry matter in ginseng leaf extract.

[0027] Figure 5 This is a schematic diagram of the desorption curve results of ginsenoside Re. Detailed Implementation

[0028] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0029] Example 1

[0030] Research on the preparation process of purslane extract suitable for chewing gum production.

[0031] The following experiments were conducted on purslane: (1) screening of anti-hypoxia active components; (2) the effect of the fineness of the powder; (3) orthogonal experiments on the extraction of medicinal materials; (4) optimization of the process conditions for centrifugation precipitation; and (5) optimization of the process conditions for precipitation drying. It was confirmed that (1) the anti-hypoxia active components were obtained by reflux extraction with ethanol, pH adjusted to neutral with 10% NaOH, and centrifugation; (2) direct feeding of purslane slices would not affect the extraction rate; (3) the optimal combination for extraction of medicinal materials was 15 times 80% ethanol, extraction once, and extraction time of 0.5h; (4) the optimal combination for centrifugation precipitation was to recover the extract until there was no ethanol, add water to 0.5 g / ml, adjust the pH to 6.5~7.0 with 10% NaOH, centrifuge at 5000 rpm for 1 min, and wash the precipitate once; and (5) the optimal drying process conditions were vacuum drying at 60℃.

[0032] (1) Screening of hypoxia-resistant active components of purslane:

[0033] 1000 g of purslane herb was extracted with 10 times its volume of 80% ethanol under reflux for 1 hour per extraction. The solvent was recovered under reduced pressure, and distilled water was added to bring the volume to 1000 ml. The pH was adjusted to neutral with 10% NaOH (A). 900 ml of this neutral concentrate was centrifuged at 5000 rpm for 5 minutes, and the precipitate was collected (B). The supernatant was replenished with water to bring the volume to 900 ml (C). 400 ml of the supernatant was extracted with petroleum ether (400 ml × 3 times), chloroform (400 ml × 3 times), and n-butanol (400 ml × 3 times), respectively, yielding the petroleum ether extract (D), chloroform extract (E), n-butanol extract (F), and the aqueous layer after extraction (G). Take another 400 ml of the supernatant after centrifugation and pass it through a D101 macroporous resin (φ50 mm × 1100 cm) for adsorption. Elute with 5000 ml of water, 50% ethanol, and 95% ethanol, respectively, to obtain the water-eluted fraction (H), the 50% ethanol-eluted fraction (I), and the 95% ethanol-eluted fraction (J). Recover the solvent from each fraction under reduced pressure until dry. Weigh an appropriate amount and suspend in 5% CMC-Na for screening of anti-hypoxia pharmacological activity. The pharmacological test methods are as follows:

[0034] Healthy male ICR mice were randomly divided into three groups according to body weight: a blank solvent control group (negative control group), a ginseng stem and leaf total saponins capsule group (positive control group), and a purslane part group, with 10 mice in each group, half male and half female. Animals were administered the drug via gavage at a volume of 0.5 ml per animal. The blank solvent control group received an equal volume of blank solvent. The administration was repeated once daily for one week. One hour after the last administration, each animal was placed in a 200 ml wide-mouth bottle containing 10 g of soda lime, one animal per bottle. The bottle opening was coated with Vaseline, sealed tightly to prevent air leakage, and then sealed again with plastic wrap. Timing was recorded immediately after sealing the bottle, and the time to death due to asphyxiation from respiratory arrest was used as the indicator. Statistical analysis was performed using SPSS for Windows 11.0 software. Data conforming to a normal distribution were used. Statistical description was performed. After testing for normality and homogeneity of variance, the quantitative data were compared among multiple groups using analysis of variance. The results showed that the total extract of purslane and the precipitate after centrifugation had significant anti-hypoxia effects, while other parts did not. The results are shown in Table 1.

[0035] Table 1. Screening results of the hypoxia resistance activity of purslane ( )

[0036]

[0037] Compared with the blank solvent control group, * P<0.05; ** P<0.01.

[0038] (2) Optimization of process conditions for extraction steps

[0039] ① The effect of the fineness of medicinal material powder

[0040] Purslane was extracted twice with 80% ethanol, once as a processed herb, once as a powder passed through sieves No. 1 and No. 2, and once as a whole, with 50 g of herb per sample. Each extraction lasted 1 hour. The extracts were combined and diluted to 1000 ml with 80% ethanol. After mixing, 5 ml of the diluted extract was evaporated to dryness, dissolved in an equal volume of methanol, and diluted to volume. The extract was then injected and the extraction rate was calculated. The results are shown in Table 2. As can be seen from the table, the difference in extraction rate among the three degrees of powdering is small. To facilitate pretreatment and filtration and simplify the process, direct extraction with processed herb slices is preferable.

[0041] Table 2. Effect of herb powder fineness on extraction rate

[0042]

[0043] ② Orthogonal Experiment for Extraction of Medicinal Materials

[0044] Based on the optimal extraction rate, an orthogonal experimental design was used to investigate the effects of ethanol concentration, extraction solvent volume, extraction time, and extraction times on the extraction effect, using extract yield and α-linolenic acid extraction rate as indicators. The results are shown in Tables 3-6. As can be seen from the tables, the effects of all investigated factors were not significant. Based on the range analysis results, considering both indicators, A2B2C1D1 was selected as the optimal combination of process conditions. Considering the texture of purslane, the optimal combination was A2B3C1D1, i.e., purslane was extracted with 15 times the concentration of 80% ethanol, once, for 0.5 h.

[0045] Table 3 Factor Level Table

[0046]

[0047] Table 4. Orthogonal experimental design and results for L9(34)

[0048]

[0049] Table 5. Analysis of Variance of Extraction Rate

[0050]

[0051] Table 6. Analysis of Variance of Extract Yield

[0052]

[0053] Under the optimal process conditions, 25g of medicinal material was taken in 3 portions, 15 times the amount of 80% ethanol, and heated under reflux once for 0.5 h each time. The results are shown in Table 7, indicating that the optimized process has good repeatability.

[0054] Table 7 Validation Test Results

[0055]

[0056] ③ Optimization of process conditions for the centrifugal sedimentation step

[0057] 300 g of purslane herb was heated under reflux for 0.5 h in 15 times the volume of 80% ethanol. The resulting extract was mixed thoroughly and divided into 12 equal portions. Nine portions were used for orthogonal experiments, and three portions were used for validation. Following the orthogonal design table, the pH was adjusted, the extract was centrifuged, the precipitate was washed with water, centrifuged again, and the precipitate was dried under reduced pressure to constant weight. The precipitate was weighed, and the yield of the extract was calculated. The extract was injected, and the linolenic acid content and extraction rate were determined. The results are shown in Tables 8-12. Based on the results of variance and range analysis of the three indicators, A2B2C1D1 was selected as the optimal combination.

[0058] Table 8 Factor Level Table

[0059]

[0060] Table 9. Orthogonal experimental design and results for L9(34)

[0061]

[0062] Table 10. Analysis of Variance Table of Transfer Rate

[0063]

[0064] Table 11 Analysis of Variance of Extract Yield

[0065]

[0066] Table 12 Analysis of variance table of α-linolenic acid content in extract

[0067]

[0068] Under the optimal process conditions A2B2C1D1, the purslane extract was recovered until ethanol was removed, water was added to 0.5 g / ml, the pH was adjusted to 6.5~7.0 with 10% NaOH, centrifuged at 5000 rpm for 1 min, and the precipitate was washed once with water. The results are shown in Table 13. The results indicate that the optimized process has good reproducibility.

[0069] Table 13 Verification Test Results

[0070]

[0071] ④ Optimization of process conditions for the precipitation and drying step

[0072] Purslane was extracted with 15 times its volume of 95% ethanol under reflux until no ethanol odor remained. Water was added to a concentration of 0.5 g / ml, and the pH was adjusted to 6.5–7.0 with 10% NaOH. The mixture was centrifuged at 5000 rpm for 1 min, and the precipitate was washed once with water. The extract was then dried using different methods. The results are shown in Table 14. With increasing drying temperature and time, the α-linolenic acid content in the extract significantly decreased. Vacuum drying produced a product with good properties, facilitating subsequent processing; therefore, vacuum drying at 60℃ was selected as the final drying process condition.

[0073] Table 14 Results of the Drying Method Investigation

[0074]

[0075] (3) Validation of small-scale intermediates

[0076] Using the optimized process conditions for preparing purslane extract, two small-scale tests were conducted with 500 g / sample of purslane raw material. The results are shown in Table 15, indicating that the optimized process conditions have good repeatability. The pharmacological results are shown in Table 16, indicating that the intermediate prepared using the optimized process has significant anti-hypoxia effects.

[0077] Table 15 Results of Small-Scale Experimental Intermediate Studies

[0078]

[0079] Table 16 Results of the anti-hypoxia pharmacological test of the intermediate in the small-scale experiment

[0080]

[0081] Compared with the blank solvent group, * P < 0.05; ** P < 0.01

[0082] (4) Process verification and scale-up experiments

[0083] Three scale-up experiments were conducted on the extraction of purslane extract under the selected process conditions. The results are shown in Table 17. This indicates that the optimized process has good repeatability and is easy to scale up for production.

[0084] Table 17 Results of three batches of scale-up experiments on purslane extract

[0085]

[0086] The optimal preparation method of purslane extract includes the following steps: purslane slices are extracted once by reflux with 15 times the volume of 80% ethanol for 0.5 h, concentrated under reduced pressure, dissolved in distilled water, and the pH is adjusted to 6.5-7.0 with 10% NaOH solution. The mixture is then centrifuged (5000 rpm for 1 min), the water is precipitated once, and the mixture is vacuum dried (at 60℃ for 8 h) to obtain the purslane extract.

[0087] Example 2

[0088] Optimization Study of Preparation Process Conditions for Ginseng Leaf Extract

[0089] Using ginsenoside Re as an indicator component, the preparation process of ginseng leaf extract was investigated. The main research contents included (1) optimization of extraction process conditions; (2) optimization of purification process conditions; and (3) process verification and scale-up experiments. The results showed that: (1) the extraction process of boiling twice with 50% ethanol for 2 h each time (keeping it at a gentle boil), filtering the extract while it was hot, combining the filtrates, concentrating under reduced pressure and adjusting the volume to 600 ml, and extracting twice was better. (2) For purification, D101 type macroporous resin can be selected as the raw material for resin purification process. It is eluted with 6 BV of water, 40% ethanol, and 95% ethanol in sequence, and the 40% ethanol eluent is collected. The resin can be maintained with 95% ethanol and recycled. It needs to be regenerated every two cycles. Regeneration was performed using acid and alkali. The regeneration method was to wash with 6 BV of 5% HCl, wash with 4000 ml of distilled water until the pH was neutral, wash with 6 BV of 5% NaOH solution, leave the column about 10 cm above the resin surface and let it stand for about 3 hours before filtering, and wash with 4000 ml of distilled water until the pH was neutral.

[0090] (1) Optimization of process conditions for extraction steps

[0091] ① Experimental Design: Ginseng leaves mainly contain ginsenosides, and the extraction method of hot reflux extraction with ethanol aqueous solution was proposed. After the initial selection of the extraction method, four factors that have a significant impact on the extraction efficiency were investigated. These factors are: A. Ethanol concentration (%); B. Amount of ethanol added (times); C. Extraction time (h); D. Number of extractions (times). The results are shown in Table 18, with three levels for each factor. Coarse ginseng leaf segments, 25 g / part, were extracted by decoction. The experiment was conducted according to the L9(34) orthogonal array. The extract was filtered while hot, the filtrates were combined, concentrated under reduced pressure and diluted to 250 ml. The extract was filtered through a microporous membrane, diluted 10 times, and subjected to HPLC analysis. The transfer rate of ginsenoside Re in the extract was used as the evaluation index. The content determination method was based on Part I of the 2005 edition of the Chinese Pharmacopoeia. The content determination results and variance analysis results are shown in Tables 19-20.

[0092] Table 18 Factor Level Table

[0093]

[0094] Table 19 L9(34) Orthogonal Experimental Design and Results

[0095]

[0096] Table 20 Analysis of Variance Table of Ginsenoside Re Transfer Rate

[0097]

[0098] ② Experimental Results: The results show that the influence of the four factors on the extraction effect of the decoction method is in the order of D > A > C > B. This indicates that the number of extractions has the greatest impact, followed by ethanol concentration, then extraction time, while the amount of ethanol used has the least impact within the tested range. Therefore, the optimal combination of levels for each factor is D2 A1 C3 B3. That is, coarse ginseng leaf segments are extracted twice using 12 times the amount of the original medicinal material with 50% ethanol, each time for 2 hours.

[0099] ③ Verification Experiment: Coarse ginseng leaf segments (25 g / sample) were decocted twice with 300 ml of 50% ethanol, 2 hours each time (maintaining a gentle boil). The extract was filtered while hot, and the filtrates were combined, concentrated under reduced pressure, and brought to a final volume of 600 ml. This process was repeated twice. After filtration through a microporous membrane, the extract was diluted 10-fold, and the content of ginsenoside Re in the extract was determined by HPLC. The results are shown in Table 21. The results indicate that the transfer rate of ginsenoside Re in the ginseng leaf extract prepared by this extraction process was 101.97%, the extract had good properties, the process was stable, and the reproducibility was good.

[0100] Table 21 Verification Test Results

[0101]

[0102] (2) Optimization of purification process conditions

[0103] The main components of ginseng leaf ethanol extract are sugars, saponins, and volatile oils. To purify ginsenosides, water-soluble sugars and other components must be removed as impurities. Macroporous adsorption resins are organic polymer adsorbents with the ability to selectively adsorb organic compounds. Based on the adsorption characteristics of macroporous resins, five types of resins—D101, HPD100, SYD-8, 860021, and DM130—were selected for optimal adsorption.

[0104] ①Preferred resin type

[0105] Sample preparation: Ginseng leaf extract was prepared using the optimized extraction process obtained from orthogonal experiments. Specifically, 2000 g of coarse ginseng leaf segments were extracted twice with 24000 ml of 50% ethanol using a decoction method, each time for 2 hours (maintaining a gentle boil). After cooling, the extract was filtered through double-layered gauze, and the filtrates were combined, concentrated under reduced pressure, and brought to a final volume of 4000 ml (crude drug concentration of 0.5 g / ml). The extract was then filtered through a microporous membrane, diluted 20-fold, and HPLC analysis revealed that the ginsenoside Re content in the extract was 0.32 mg / ml. This mixed extract was used as the sample solution for the resin purification process.

[0106] ② Resin pretreatment:

[0107] Five types of resins—D101, HPD100, SYD-8, 860021, and DM130—were packed into columns. The columns were then immersed in 95% ethanol at a depth of 10-20 cm above the resin layer for 3-4 hours, followed by draining the washing solution. The column was then washed with 6 BV of 95% ethanol until the effluent did not become cloudy when three times the volume of water was added to the test tube. The column was then thoroughly washed with water until no alcohol odor remained.

[0108] ③ Experimental methods and results

[0109] After thoroughly drying the treated resin, weigh 5g into a 250ml Erlenmeyer flask, add 100ml of the above-mentioned solution, and shake by hand once every 2 hours. After 24 hours (equivalent to 12 hours of shaking with a shaker), determine the static adsorption effect of the resin. Calculate the adsorption capacity for different amounts using the following formula:

[0110] Q=(C0-C e )×V / w

[0111] In the formula: C0 is the concentration of the solution before adsorption (mg / ml), C e denoted as , where is the concentration of the solution after adsorption (mg / ml), w is the weight of the resin (g), V is the volume of the solution (ml), and Q is the amount of adsorption.

[0112] The statically adsorbed resin was blotted dry with paper, and 50 ml of 70% ethanol was added to each bottle. The mixture was shaken once every 2 hours. After 24 hours, the desorption effect of the resin was measured, and the resolution rate of different types of resin was calculated. The experimental results are shown in Table 22. The results indicate that among the five resins investigated, the D101 macroporous resin had the strongest adsorption capacity for ginsenosides Rg1 and Re. Although its resolution rate was slightly lower than that of HPD100 and SYD-8 resins, the D101 macroporous resin was selected as the preferred raw material for the resin purification process after comprehensive consideration.

[0113] Table 22 Optimal Selection Results of Different Types of Macroporous Resins

[0114]

[0115] ④Performance evaluation of D101 macroporous adsorption resin for the purification of ginseng leaf extract

[0116] Sample solution preparation: Take the above-mentioned sample solution with a crude drug concentration of 0.5 g / ml and dilute it with distilled water to prepare test sample solutions with crude drug concentrations of 0.4, 0.2, 0.1, 0.05, and 0.025 g / ml, respectively.

[0117] Resin column packing and pretreatment: Based on the resin optimization test results, 100 g of D101 macroporous resin was weighed and packed into a column (4cm×16cm) with a column volume of 140 ml and a dead volume of 30 ml. The resin was then pretreated according to the above-mentioned resin pretreatment method and set aside for later use.

[0118] Effect of sample solution concentration on adsorption: Adsorption experiments were conducted using sample solutions of different concentrations at a flow rate of 1.5 BV / h. The content of ginsenoside Re in the effluent was measured. Sample addition was stopped when the concentration reached 0.2 mg / ml. The maximum adsorption capacity under each concentration condition was calculated. The results are shown in [Figure Number]. Figure 1 As shown, Figure 1 This is a schematic diagram showing the relationship between sample solution concentration and resin adsorption capacity. As can be seen from the figure, sample solution concentration has a significant impact on the resin's adsorption performance. When the sample solution concentration is 0.4 g / m³, the D101 macroporous adsorption resin exhibits the highest adsorption capacity for ginsenoside Re.

[0119] Qmax = C × Vmax

[0120] In the formula: C is the concentration of the loading solution (g / ml), Vmax is the maximum loading volume (ml), and Qmax is the maximum adsorption capacity.

[0121] Adsorption capacity: A sample solution with a crude drug concentration of 0.4 g / ml was passed into a D101 resin column (4 cm × 16 cm, column volume 140 ml) at a flow rate of 1.5 BV / h. The content of ginsenoside Re in the eluent was measured until the resin was saturated. Results are shown below. Figure 2 As shown, Figure 2 This is a schematic diagram of the adsorption curve results of ginsenoside Re from ginseng leaves on D101 type resin. The results show that 100 g of D101 macroporous adsorption resin can process approximately 9 / 7 BV of sample solution when saturated. After processing to 8 / 7 BV, the concentration of ginsenoside Re in the effluent is 0.03 mg / ml, and after processing to 9 / 7 BV, the concentration is 0.38 mg / ml. Since the concentration exceeds 0.2 mg / ml, the sample loading amount exceeds the resin's maximum loading capacity. To reduce losses, the actual sample loading amount during purification was determined to be 8 / 7 BV.

[0122] Effect of flow rate on adsorption: A sample of crude drug with a concentration of 0.4 g / ml was passed into a chromatographic column packed with 100 g D101 resin (4 cm × 16 cm, column volume 140 ml) at different flow rates. The flow was stopped when the concentration of ginsenoside Re in the effluent exceeded 0.2 mg / ml. The effect of flow rate on resin adsorption was investigated by calculating the maximum adsorption capacity under different flow rate conditions. The calculation method for the maximum adsorption capacity was the same as above. Results are shown below. Figure 3 As shown, Figure 3 This is a schematic diagram showing the relationship between flow rate and sample adsorption capacity. The results show that the resin has a larger adsorption capacity at lower flow rates, and the adsorption capacity decreases with increasing flow rate. Flow rates of 1.5 BV / h and 1.0 BV / h have similar adsorption capacities. Considering the time factor, a flow rate of 1.5 BV / h was selected.

[0123] Desorption performance of D101 macroporous adsorption resin: Considering the safety of the drug and the physicochemical properties of ginsenosides, an ethanol-water system was selected as the optimal eluent. 160 ml of the test sample solution with a crude drug concentration of 0.4 g / ml was passed into a D101 resin column (4 cm × 16 cm, column volume 140 ml). Elution was performed using a 6 BV volume gradient of distilled water and ethanol of different concentrations. The eluent fractions were collected and refluxed under reduced pressure to a small volume, then dried under reduced pressure at 60℃. The yield of each eluted fraction and the content of ginsenoside Re were calculated. The results are shown in Table 23. Figure 4 , Figure 5 As shown.

[0124] Table 23. Material distribution in elution sites of different ethanol concentrations

[0125]

[0126] Figure 4 This is a schematic diagram of the desorption curve results of dry matter in ginseng leaf extract. Figure 5This is a schematic diagram of the desorption curve results for ginsenoside Re. The results show that using a gradient elution of 6 BV water, 10%, 20%, 30%, 40%, 50%, 70%, and 95% ethanol, followed by concentration under reduced pressure, drying, and weighing of the eluent, the cumulative yield of ginsenoside Re from 10% to 40% ethanol was calculated to be 21.17%, indicating a good purification effect. Based on these results, the experimental protocol was adjusted. 160 ml of the test sample solution with a concentration of 0.4 g / ml was passed into a D101 resin column (4 cm × 16 cm, column volume 140 ml), and eluted with a gradient of 6 BV each of water, 40%, and 95% ethanol. The eluent was collected, refluxed under reduced pressure to a small volume, and dried under reduced pressure at 60℃. The yield of each eluted fraction and the content of ginsenoside Re were calculated, and the results are shown in Table 24. The results showed that after continuous elution with 6 BV of water, 40% ethanol, and 95% ethanol, the 40% ethanol eluent was concentrated under reduced pressure and dried to obtain ginseng extract with a ginsenoside Re content of 23.78%, indicating good purification effect. Therefore, the desorption method for ginsenoside Re was determined to be sequential elution with 6 BV of water, 40% ethanol, and 95% ethanol, collecting the 40% ethanol eluent.

[0127] Table 24. Material distribution in elution fractions of ethanol at different concentrations.

[0128]

[0129] ⑤ Resin maintenance, regeneration, and service life

[0130] Resin maintenance: Pass 160 ml of the test sample solution with a crude drug concentration of 0.4 g / ml into a chromatography column packed with 100 g (140 ml volume, 4 cm × 16 cm) of resin. The sample loading volume is 160 ml (8 / 7 BV). Elute with a gradient of distilled water, 40%, and 95% ethanol. Wash with 95% ethanol until the eluent is colorless and clear, and then wash with distilled water until colorless and odorless before proceeding to the next round of sample loading and elution. The resin can be recycled.

[0131] Resin regeneration: There have been many research reports on resin regeneration. Based on the resin regeneration methods recommended by resin manufacturers and combined with actual production, it has been determined that acid and alkali treatment will be used for regeneration.

[0132] The resin to be regenerated was washed with distilled water until no alcohol odor remained, and then regenerated. 6 BV of 5% HCl was directly passed through the resin column for washing, followed by washing with 4000 ml of distilled water until the pH was neutral. Then, it was washed with 6 BV of 5% NaOH solution, leaving approximately 10 cm above the resin surface on the column. After standing for about 3 hours, it was filtered out and washed with 4000 ml of distilled water until the pH was neutral.

[0133] Usage cycle: 160 ml of a test solution with a concentration of 0.4 g (raw material) / ml was passed into a chromatography column packed with 100 g (140 ml volume, 4 cm × 16 cm) of resin. The sample loading volume was 160 ml (8 / 7 BV) (equivalent to a feed volume of 64 g). Elution was performed using a gradient of 6 BV with distilled water, 40% ethanol, and 95% ethanol. The eluent was then washed with 95% ethanol until it was colorless and clear, followed by washing with distilled water until colorless and odorless. The resin was then recycled. The 40% ethanol eluent was dried, weighed, and sampled for analysis of ginsenoside Re content. The results showed that the resin adsorption rate decreased significantly after two cycles, requiring regeneration. Therefore, the regeneration cycle of the resin was determined to be two cycles.

[0134] (3) Process verification and scale-up experiments

[0135] Two small-scale tests and three large-scale tests were conducted on ginseng leaf extract under the selected process conditions. The results are shown in Tables 25 and 26. This indicates that the optimized process has good repeatability and strong operability.

[0136] Table 25 Results of the two small-scale trials

[0137]

[0138] Table 26 Results of three batches of scale-up experiments on ginseng leaf extract

[0139]

[0140] The optimal preparation method of the ginseng leaf extract includes the following steps:

[0141] The coarse ginseng leaf segments were decocted twice with 12 times the amount of 50% ethanol for 2 hours each time. The filtrates were then combined, concentrated under reduced pressure, and water was added. The mixture was then filtered through a microporous membrane, diluted, and purified to obtain the ginseng leaf extract.

[0142] Example 3

[0143] Research on the processing technology and formulation of anti-fatigue chewing gum.

[0144] To develop anti-fatigue chewable fruit balls, which are used to prevent or reduce fatigue to a certain extent, refresh the mind, and relieve stress and fatigue. The research content mainly includes three parts: (1) optimization of the edibility and main flavor of raw materials; (2) optimization of flavor blending; (3) optimization of chewing time. The research results show that: (1) Yunnan olive has a sweet aftertaste and has good synergy with ginseng leaf extract. Therefore, Yunnan olive fruit powder is selected as the main ingredient to make Yunnan olive fruit balls. (2) It is determined that the bitterness of the product with added steviol glycosides is significantly reduced. Under the sweet aftertaste of Yunnan olive, the product is bitter at first and then sweet. (3) The fruit balls are designed to be 3g / piece. According to the content requirements of ginseng leaf extract and purslane extract, the content of the two extracts in each piece is halved. Each time, consuming 2 fruit balls can achieve the content of functional ingredients. The chewing time of 2 pieces can reach more than 3 minutes, and the bitterness of the product is significantly reduced, greatly improving the taste.

[0145] (1) Experimental objective

[0146] Consuming anti-fatigue chewing gum can prevent or reduce fatigue to some extent, refresh the mind, and relieve stress and fatigue.

[0147] (2) Product technical requirements

[0148] ① Generally, each pill weighs ≤5g and the volume should be minimized as much as possible, but adjustments can be made according to the actual situation;

[0149] ② Functional ingredient content: Each piece of chewing gum contains 0.5g of purslane extract and 0.34g of ginseng leaf extract;

[0150] ③ The flavor needs to be adjusted to improve the taste;

[0151] ④ The product should be chewed continuously for no less than 3 minutes (the chewing time can be increased by adjusting the hardness or colloid content).

[0152] ⑤ It can be swallowed directly after chewing, leaving no residue;

[0153] ⑥ The product is resistant to low temperatures and can withstand special battlefield environments such as high salt and high humidity through its outer packaging. It is also stable and has a shelf life of at least 18 months.

[0154] (3) Experimental materials

[0155] Purslane extract, ginseng leaf extract, Yunnan olive fruit powder, tamarind fruit powder, passion fruit powder, duoyi fruit powder, white sugar, maltodextrin, glucose powder, steviol glycosides, gelatin, locust bean gum, Prussian polysaccharide, carrageenan, gellan gum, pectin, flaxseed gum, etc.

[0156] (4) Analysis of key points for process optimization

[0157] ① Edibility of raw materials. According to the product's technical requirements, chewing gum needs to be swallowed directly without leaving any residue. Therefore, the ingredients should be edible and swallowable food raw materials.

[0158] ② Flavor Harmony. Because the bitterness of ginseng leaf and purslane extracts is too strong, making it difficult to chew slowly, the flavor of the product needs to be harmonized and optimized.

[0159] ③ Chewing time. Chewing is an important way to combat fatigue, so the chewing time of the product needs to be optimized.

[0160] (5) Process optimization process

[0161] ① Optimization of raw material edibility and main flavor

[0162] Based on the principle of flavor synergy, natural fruit powders with a sour and astringent taste, such as tamarind fruit powder, Yunnan olive fruit powder, passion fruit powder, and Duoyi fruit powder, were selected as the main ingredients. These were then combined with ginseng leaf extract (RSE), purslane extract (MZXE), and excipients such as white sugar and maltodextrin to prepare chewing gum samples. Ginseng leaf extract and purslane extract were added according to the required content for each product.

[0163] Tests showed that Yunnan olives, due to their sweet aftertaste, exhibit good synergy with ginseng leaf extract. Therefore, Yunnan olive fruit powder was selected as the main ingredient to produce Yunnan olive fruit balls. The results are shown in Table 27.

[0164] Table 27 Optimization Results of Main Ingredients for Chewing Gum

[0165]

[0166] ② Flavor blending and optimization

[0167] The bitterness of functional ingredient extracts can be reduced by adding aromatic or flavorful ingredients. Coffee extract, mint, and milk powder were selected as ingredients to balance the bitterness. Tests showed that the cooling sensation of mint has a certain buffering and alleviating effect on the bitterness in the mouth. However, because mint itself has a certain stimulating and anti-fatigue effect, this process was not used to balance the bitterness based on product requirements.

[0168] Furthermore, natural sweeteners such as monk fruit concentrate, monk fruit glycosides, and steviol glycosides were selected to balance the bitterness. Tests confirmed that the addition of steviol glycosides significantly reduced the bitterness of the product, as shown in Table 28. Under the induction of the sweet aftertaste of Yunnan olives, the product was initially bitter but then developed a sweet aftertaste.

[0169] Table 28 Results of sweetener optimization in Yunnan olive chewing gum (3g / piece)

[0170]

[0171] ③ Optimization of chewing time

[0172] To prolong the chewing time of Yunnan olive chewing gum, food-grade hydrocolloids were added to the basic formula to improve the texture and extend the chewing time. Gelatin, locust bean gum, Prussian polysaccharide, carrageenan, gellan gum, pectin, and flaxseed gum were selected for addition to improve the gum's hardness and chewing time. Comparative testing showed that the sample with added gelatin performed best, exhibiting a chewy texture and extended chewing time, although the chewing time did not reach 3 minutes per gum piece. The results are shown in Table 29.

[0173] Therefore, the fruit balls were designed with a weight of 3g / piece. Based on the content requirements of ginseng leaf extract and purslane extract, the content of the two extracts in each piece was halved. Consuming 2 fruit balls each time can achieve the content of functional ingredients. The chewing time of 2 pieces can reach more than 3 minutes, and the bitterness of the product is significantly reduced, greatly improving the taste.

[0174] Table 29 Optimization Results of Hydrocolloid Addition in Yunnan Olive Chewing Gum (3g / piece)

[0175]

[0176] (6) Final product formula

[0177] Table 30. Yunnan Olive Chewing Gum (3g / piece) Product Formula

[0178]

[0179] Recommended dosage: 2 capsules each time.

[0180] Weigh out the following components according to the specified ratio: Yunnan olive extract, white sugar, maltodextrin, purslane extract, ginseng leaf extract, glucose powder, gelatin, coating colorant, and steviol glycosides. Mix them evenly, add water, heat and stir until completely dissolved, shape the mixture, and coat it to obtain the anti-fatigue chewing gum.

[0181] (7) Product Description

[0182] Product net weight: 3g / capsule, per-capsule tolerance ±9%. It is in the shape of small spheres, with a light blue to blue outer skin and a dark brown interior; it has the taste and aroma of Yunnan olive and ginseng leaf, without any off-odors.

[0183] Example 4

[0184] Clinical research: Study on the effectiveness of anti-fatigue chewing gum.

[0185] The experimental design and results are as follows:

[0186] (1) Experimental design:

[0187] A single-blind, placebo-controlled parallel trial was conducted to investigate the efficacy of chewing gum in relieving mental fatigue.

[0188] (2) Inclusion criteria for subjects:

[0189] ① TU students with a Mental Fatigue Scale (MFS) score >10.5 and whose study / work intensity has not changed significantly within a week; ② No age or gender restrictions, no major organic diseases, and no limb or joint injuries; ③ Willing to actively cooperate with all aspects of this survey and promise the authenticity of the subjective survey content; ④ Sign an informed consent form.

[0190] (3) Exclusion criteria:

[0191] ① There are foreseeable work assignments or transfers in the near future that will prevent the completion of all testing content in this study; ② For various reasons, medications that may affect the observed indicators may be taken concurrently during the trial; ③ There are recent illnesses such as colds that may cause secondary fatigue; ④ There is an allergy to any of the ingredients in this chewing gum.

[0192] (4) Test plan:

[0193] Subjects were randomly divided into an experimental group and a control group. The experimental group was given the anti-fatigue chewing gum prepared according to Example 3 of this invention (Table 30), while the control group was given non-medicated chewing gum (made from starch and dextrin instead of ginseng leaf and purslane extracts, with other excipients unchanged, and with the same appearance as the medicated chewing gum), chewed orally once a day, 2 pieces each time. This was continued for 1 week. The difference in fatigue levels before and after chewing the gum was statistically analyzed between the experimental and control groups to evaluate the effectiveness of the medicated chewing gum in relieving fatigue.

[0194] (5) Evaluation index: MFS mental fatigue scale score. The score was measured before the start of the experiment and after the last dose of medication.

[0195] (6) The test results are shown in Table 31:

[0196] Table 31

[0197]

[0198] The difference between the MFS score after the last use of the chewing gum and the MFS score before medication was used as the basis for efficacy evaluation. The results showed that the MFS difference in the experimental group was 5.3±2.19, and the MFS difference in the control group was 2.1±1.72. The difference between the groups was statistically significant after independent samples t test (t=3.287, p=0.005). The decrease in MFS in the experimental group was significantly higher than that in the control group. That is, the anti-fatigue chewing gum prepared in Example 3 of this invention (Table 30) has a greater ability to reduce the MFS score of users than the placebo.

[0199] The innovation of this invention lies in the fact that purslane is a herbaceous medicinal material. Water extraction yields a high extract rate, but subsequent processing is difficult, making formulation challenging. Based on a comprehensive analysis of previous research results and literature, the approach chosen was to extract, purify, and then mix ginseng leaves and purslane for formulation. An orthogonal experiment was designed to investigate factors such as extraction solvent concentration, solvent volume, extraction time, and extraction times, using α-linolenic acid extraction rate and extract yield as evaluation indicators. Analysis of variance was used to optimize and determine the optimal extraction conditions for purslane. Based on previous pharmacodynamic screening results, the purification step for purslane adopted an alcohol extraction and water precipitation process. Specific process parameters, including mother liquor concentration, pH, centrifugation time, and number of water washes, were determined through orthogonal experiments. Validation of this process showed that the obtained extract achieved the goals of ensuring efficacy, maximizing impurity removal, and minimizing solids yield. Scale-up of the process demonstrated its simplicity, stability, good reproducibility, and high safety. In the drying step of the extract, by comparing the differences between atmospheric pressure drying and vacuum drying in terms of drying efficiency, appearance of dried product, and impact on active ingredients, vacuum drying was determined to be the preferred method.

[0200] This invention relates to the extraction of ginseng leaves. Based on references and previous research results, an orthogonal experiment was designed to investigate factors such as extraction solvent concentration, solvent volume, extraction time, and extraction times. The extraction rate of ginsenoside Re was used as the evaluation index. The results were analyzed using variance to optimize and determine the optimal extraction conditions for ginseng leaves. In the purification step, referring to existing and relatively mature macroporous resin purification processes, the process parameters, including resin type, loading concentration, loading speed, loading volume, and elution volume, were systematically optimized.

[0201] Based on the principle of flavor synergy, this invention selects Yunnan olive fruit powder as the main ingredient from natural fruit powders with a sour and astringent taste, such as tamarind fruit powder, Yunnan olive fruit powder, passion fruit powder, and Duoyi fruit powder; selects steviol glycosides from monk fruit concentrate, monk fruit glycosides, and steviol glycosides to balance bitterness; and selects gelatin from gelatin, locust bean gum, Prussian polysaccharide, carrageenan, gellan gum, pectin, and flaxseed gum to extend chewing time. However, even the sample with added gelatin showed the best results, with a chewy texture and extended chewing time, but the chewing time did not reach 3 minutes per capsule. Therefore, a 3g / capsule fruit ball was designed. Based on the content requirements of ginseng leaf extract and purslane extract, the content of both extracts in each capsule was halved. Consuming 2 fruit balls at a time achieves the required content of functional ingredients, and the chewing time of 2 capsules can reach more than 3 minutes, with a significant reduction in bitterness and a greatly improved taste.

[0202] Example 5

[0203] A method for preparing an anti-fatigue chewing gum includes the following steps:

[0204] The anti-fatigue chewing gum is made from the following ingredients by weight percentage: 45.6% Yunnan olive extract, 21.3% white sugar, 13.2% maltodextrin, 10.88% purslane extract, 5% ginseng leaf extract, 2.8% glucose powder, 1% gelatin, 0.2% coating colorant, and 0.02% steviol glycosides.

[0205] Weigh out the following components according to the specified ratio: Yunnan olive extract, white sugar, maltodextrin, purslane extract, ginseng leaf extract, glucose powder, gelatin, coating colorant, and steviol glycosides. Mix them evenly, add water, heat and stir until completely dissolved, shape the mixture, and coat it to obtain the anti-fatigue chewing gum.

[0206] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A method of preparing a fatigue-resistant chewing gum, characterized in that, Includes the following steps: The anti-fatigue chewing gum is made from the following components by weight percentage: 40-50% Yunnan olive extract, 20-25% white sugar, 8-16% maltodextrin, 5-10% purslane extract, 2-8% ginseng leaf extract, 1-5% glucose powder, 0.1-2% gelatin, 0.01-1% coating colorant, and 0.005-0.1% steviol glycosides. Weigh out the following components according to the specified ratio: Yunnan olive extract, white sugar, maltodextrin, purslane extract, ginseng leaf extract, glucose powder, gelatin, coating colorant, and steviol glycosides. Mix them evenly, add water, heat and stir until completely dissolved, shape the mixture, and coat it to obtain the anti-fatigue chewing gum.

2. The method of making a fatigue-resistant chewing gum according to claim 1, wherein, The anti-fatigue chewing gum is made from the following components by weight percentage: 47.4% Yunnan olive extract, 22.48% white sugar, 12% maltodextrin, 8.3% purslane extract, 5.6% ginseng leaf extract, 3% glucose powder, 1% gelatin, 0.2% coating colorant, and 0.02% steviol glycosides.

3. The method of claim 1, wherein the anti-fatigue chewing gum is prepared by the steps of: The anti-fatigue chewing gum is made from the following components by weight percentage: 45.6% Yunnan olive extract, 21.3% white sugar, 13.2% maltodextrin, 10.88% purslane extract, 5% ginseng leaf extract, 2.8% glucose powder, 1% gelatin, 0.2% coating colorant, and 0.02% steviol glycosides.

4. The method of claim 1, wherein the anti-fatigue chewing gum is prepared by the steps of: The preparation method of the ginseng leaf extract includes the following steps: Add 8-14 times the volume of 40-95% ethanol to the coarse ginseng leaf segments and decoct 1-3 times for 0.5-2 hours. Filter, combine the filtrates, concentrate under reduced pressure, add water, filter through a microporous membrane, dilute, and purify to obtain the ginseng leaf extract.

5. The method of claim 4, wherein the anti-fatigue chewing gum is prepared by the steps of: The preparation method of the ginseng leaf extract includes the following steps: The coarse ginseng leaf segments were decocted twice with 12 times the amount of 50% ethanol for 2 hours each time. The filtrates were then combined, concentrated under reduced pressure, and water was added. The mixture was then filtered through a microporous membrane, diluted, and purified to obtain the ginseng leaf extract.

6. A process for the preparation of a fatigue-resistant chewing gum according to claim 4 or 5, characterized in that, The purification was performed using a D101 macroporous resin column, eluting sequentially with water, 40% ethanol, and 95% ethanol, and collecting the 40% ethanol eluent.

7. The method of claim 1, wherein the anti-fatigue chewing gum is prepared by the steps of: The preparation method of the purslane extract includes the following steps: purslane slices are extracted by reflux with 8-20 times 50-95% ethanol, concentrated under reduced pressure, dissolved in distilled water, and the pH is adjusted to 6.5-7.0 with 5-15% NaOH solution. The mixture is then centrifuged, the precipitate is washed with water, and vacuum dried to obtain the purslane extract.

8. The method of claim 1, wherein the anti-fatigue chewing gum is prepared by the steps of: The preparation method of the purslane extract includes the following steps: purslane slices are extracted once by reflux with 15 times 80% ethanol for 0.5 h; the extraction is concentrated under reduced pressure, dissolved in distilled water, and the pH is adjusted to 6.5-7.0 with 10% NaOH solution. The mixture is then centrifuged at 5000 rpm for 1 min, the precipitate is washed once with water, and then vacuum dried at 60℃ for 8 h to obtain the purslane extract.

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