Shandong colla corii asini pulp composition with effect of preventing excessive internal heat and application thereof

Abstract

The invention provides a Shandong colla corii asini pulp composition without causing internal heat and a preparation process thereof. The composition comprises colla corii asini, fructus lycii, radix rehmanniae praeparata, radix astragali and fructus ziziphi jujubae. The invention also provides a preparation method and application of the composition. According to the composition, traditional Chinese medicinal materials and donkey-hide gelatin are matched, so that the blood replenishing and nourishing effects of the donkey-hide gelatin are enhanced, discomfort caused by poor taste when part of people eat the donkey-hide gelatin is solved, the dryness of the donkey-hide gelatin is relieved, the risk of excessive internal heat after eating is reduced, and meanwhile, the Shandong donkey-hide gelatin pulp prepared by adopting the preparation method is fine and smooth in taste, delicious in taste and free of toxic and side effects. The product is suitable for people with various constitutions.

Description

Technical Field

[0001] This invention relates to the field of donkey-hide gelatin technology, specifically to a Shandong donkey-hide gelatin syrup composition that does not cause internal heat and its application. Background Technology

[0002] The statements herein provide only background information in relation to this invention and do not necessarily constitute prior art.

[0003] Animal-derived medicines such as donkey-hide gelatin often have an unpleasant taste and are difficult to swallow. Furthermore, due to its rich and greasy nature, donkey-hide gelatin can easily cause indigestion. In addition, due to factors such as constitution, emotions, lifestyle, and method of consumption, "heatiness" often occurs when consuming donkey-hide gelatin.

[0004] Traditional methods of consuming donkey-hide gelatin include melting it, stewing it in rice wine, and making donkey-hide gelatin cakes, but none of these methods effectively address the issue of "heatiness" caused by its consumption. Therefore, there is an urgent need to invent a method of consuming donkey-hide gelatin that does not cause heatiness, resulting in a product that not only tastes good but also has excellent blood-nourishing effects. This method utilizes traditional Chinese medicine formulations to enhance the effects of donkey-hide gelatin in improving nutritional anemia while avoiding the "heatiness" phenomenon caused by its consumption. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the purpose of this invention is to provide a Shandong donkey-hide gelatin syrup composition that does not cause internal heat and its application. This invention utilizes donkey-hide gelatin and traditional Chinese medicine extracts through stewing and other methods to reduce the dryness of the donkey-hide gelatin, thus preventing internal heat and adjusting its taste. The use of traditional Chinese medicine enhances the efficacy of donkey-hide gelatin in improving nutritional anemia while avoiding the "internal heat" phenomenon caused by consuming donkey-hide gelatin.

[0006] The present invention achieves the above objectives through the following technical solutions: In a first aspect, the present invention provides a Shandong donkey-hide gelatin oral liquid composition and its preparation process that does not cause internal heat, comprising donkey-hide gelatin, wolfberry, prepared rehmannia root, astragalus root, and jujube.

[0007] The preferred ingredients are: 50-70g of donkey-hide gelatin, 30-50g of wolfberry, 70-90g of prepared rehmannia root, 15-25g of astragalus root, and 90-110g of jujube.

[0008] A more preferred combination is 60g of donkey-hide gelatin, 40g of wolfberry, 80g of prepared rehmannia root, 20g of astragalus root, and 100g of jujube.

[0009] Its preparation process includes, but is not limited to, the following steps: The ethanol percolation method was used to extract wolfberry, astragalus and jujube, and the percolate was concentrated to obtain percolate concentrate. The prepared Rehmannia root was extracted using water extraction. The extracted water extract and the percolated concentrate were subjected to cold precipitation. The precipitate was filtered and concentrated to obtain the water-extracted percolated concentrate. The above-mentioned water-extracted percolate, donkey-hide gelatin solution, and sweetener are prepared once, heated, cooled and filtered, stabilized, heated a second time, cooled and filtered, and the cooled sediment is centrifuged, bottled, and sterilized.

[0010] In some implementation schemes, the percolation process for wolfberry, astragalus, and jujube involves using 60% ethanol as a solvent. Specifically, before percolation, atractylodes macrocephala and wolfberry are soaked in a solvent 8 to 10 times the volume of the herbs, and the percolation time is 24 hours.

[0011] In some implementation schemes, the water extraction process of Rehmannia glutinosa involves: decocting Rehmannia glutinosa with water to obtain a decoction, and then filtering the decoction to obtain a filtrate. Specifically, the decoction is performed at least three times, with each decoction lasting 1.5 to 2.5 hours, and the decoctions are then combined.

[0012] In some implementation schemes, the water extraction process of Rehmannia glutinosa is as follows: Rehmannia glutinosa is soaked in 8 times its volume of water for 0.5 hours, heated to 80-85℃, and subjected to low-temperature ultrasonic extraction at 200W and 40kHz for 2 hours. Ultrasonic extraction disrupts the cell walls of Rehmannia glutinosa, promoting the dissolution of yin-nourishing polysaccharides while avoiding the degradation of components and the generation of heat-inducing derivatives due to high temperatures. The decoction is prepared at least three times, and the decoctions are combined. First, suspended impurities are removed by coarse filtration through gauze, and then purified sequentially through a three-stage ultrafiltration membrane: a first-stage ultrafiltration membrane (20000 Da) removes large molecular weight proteins and tannins; a second-stage ultrafiltration membrane (10000 Da) removes residual large molecular weight polysaccharides; and a third-stage ultrafiltration membrane (5000 Da) further purifies the solution. The permeate from the third-stage ultrafiltration is collected, and 0.1% citric acid is added to adjust the pH to 5.5-6.0 to obtain the Rehmannia glutinosa decoction.

[0013] Specifically, the preferred steps of the preparation process are: 1. Soak wolfberries, astragalus, and jujubes in 0.8 times their volume of 60% ethanol at room temperature for 0.5 hours. Then, add 60% ethanol to completely submerge the surface of the herbs and soak for 24 hours. After soaking, percolate. The total amount of 60% ethanol used in soaking, maceration, and percolation should be 10 times the volume of the herbs. Let the percolate stand for 4 hours, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.15-1.20 (80℃). Set aside for later use. 2. Soak Rehmannia glutinosa in 8 times its volume of water for 0.5 hours, then heat to 80-85℃ and extract using low-temperature ultrasonication at 200W and 40kHz for 2 hours. Filter the decoction. Extract the residue twice, adding 4 times and 3 times the volume of water respectively, for 1.5 hours each time. Combine the decoctions. First, use gauze to coarsely filter and remove suspended impurities, then purify it sequentially through a three-stage ultrafiltration membrane: 20000Da for the first stage, 10000Da for the second stage, and 5000Da for the third stage. Collect the permeate from the third stage and add 0.1% citric acid to adjust the pH to 5.5-6.0 to obtain the Rehmannia glutinosa decoction. This process inhibits the dissolution of impurities that may cause internal heat and improves the effect of cold precipitation for impurity removal. 3. Add the Rehmannia glutinosa decoction to the above permeate concentrate, stir for 30 minutes, and allow it to settle at 2-10℃ for 48 hours. Filter the solution and concentrate the filtrate under reduced pressure to a relative density of 1.05-1.10 (80℃). 4. Grind the donkey-hide gelatin into powder, weigh accurately, add 6 times the amount of purified water, stir until completely dissolved, keep simmering for 30 minutes, filter, and set aside. 5. Add an appropriate amount of white sugar to 4 times the amount of water, boil until dissolved, and set aside as a sugar syrup. 6. Mix the concentrated extract of medicinal slices, the enzymatic hydrolysate of donkey-hide gelatin, and the sugar solution, stir for 30 minutes, boil for 30 minutes, cool and settle at 2~8℃ for 48 hours, centrifuge, add water to the specified volume of the centrifuged liquid, centrifuge again, fill and sterilize.

[0014] Secondly, this invention provides a Shandong donkey-hide gelatin oral composition that does not cause internal heat, and its application in improving nutritional anemia.

[0015] The beneficial effects achieved by one or more embodiments of the present invention are as follows: This invention provides a composition of Shandong donkey-hide gelatin syrup that does not cause internal heat. This composition combines Chinese medicinal herbs with donkey-hide gelatin, which not only enhances the blood-nourishing effect of donkey-hide gelatin, but also solves the discomfort caused by the unpleasant taste of donkey-hide gelatin for some people, and alleviates the dryness of donkey-hide gelatin itself, reducing the risk of "internal heat" after consumption. At the same time, the Shandong donkey-hide gelatin syrup prepared by this method has a delicate taste and delicious flavor, making it suitable for people of all constitutions. Attached Figure Description

[0016] Figure 1 This study compares the fecal samples of mice in different groups, including the normal control group, the low-dose group of Shandong donkey-hide gelatin syrup, the medium-dose group of Shandong donkey-hide gelatin syrup, the high-dose group of Shandong donkey-hide gelatin syrup, the compound donkey-hide gelatin syrup group, the ginseng group, and the red ginseng and astragalus group. Figure 2 This study compares the urine of mice in different groups, including the normal control group, the low-dose group of Shandong donkey-hide gelatin oral liquid, the medium-dose group of Shandong donkey-hide gelatin oral liquid, the high-dose group of Shandong donkey-hide gelatin oral liquid, the compound donkey-hide gelatin oral liquid group, the ginseng group, and the red ginseng and astragalus group. Figure 3This study compares the tongue appearance of mice in different groups, including the normal control group, the low-dose group of Shandong donkey-hide gelatin syrup, the medium-dose group of Shandong donkey-hide gelatin syrup, the high-dose group of Shandong donkey-hide gelatin syrup, the compound donkey-hide gelatin syrup group, the ginseng group, and the red ginseng and astragalus group. Detailed Implementation

[0017] It should be noted that the following detailed description is illustrative and intended to provide further explanation of the invention. Unless otherwise specified, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0018] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of the present invention.

[0019] Experimental Example 1: Comparative Study of Shandong Donkey-hide Gelatin Syrup Processing Technology Experiment Example 2: Efficacy of Shandong Donkey-hide Gelatin Oral Liquid and Preparation of "Non-Heat-Inducing" Samples This experiment used donkey-hide gelatin produced within the past year to prepare Shandong donkey-hide gelatin syrup. The preparation process is as follows: 1. Soak 400g of wolfberry, 200g of astragalus, and 1000g of jujube in 1280mL of 60% ethanol at room temperature for 0.5 hours. Then, add 60% ethanol to submerge the surface of the herbs and soak for 24 hours. Percolate the mixture. The total amount of 60% ethanol used in soaking, maceration, and percolation is 1600mL. Let the percolate stand for 4 hours, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.18 (80℃). Set aside for later use. 2. Soak 800g of Rehmannia glutinosa in 6400mL of water for 0.5 hours, then heat and extract for 2 hours. Filter the decoction. Extract the residue twice, adding 3200mL and 2400mL of water respectively, for 1.5 hours each time. Combine the decoctions. First, use gauze to coarsely filter and remove suspended impurities, then purify it by passing it through a three-stage ultrafiltration membrane: 20000Da for the first stage, 10000Da for the second stage, and 5000Da for the third stage. Collect the permeate from the third stage ultrafiltration, add 0.1% citric acid to the permeate, and adjust the pH to 5.5-6.0 to obtain the Rehmannia glutinosa decoction. 3. Add the decoction of Rehmannia glutinosa to the above percolation concentrate, stir for 30 minutes, cool and settle at 5°C for 48 hours, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.07 (80°C). 4. Grind 600g of donkey-hide gelatin into coarse powder, weigh accurately, add 3600mL of purified water, stir until completely dissolved, keep simmering for 30 minutes, filter, and set aside. 5. Add 1400g of white sugar to 5600mL of water, boil and dissolve to obtain sugar syrup, set aside; 6. Mix the concentrated extract of medicinal slices, the enzymatic hydrolysate of donkey-hide gelatin, and the sugar solution, stir for 30 minutes, boil for 30 minutes, cool and settle at 2~8℃ for 48 hours, centrifuge, add water to the centrifuged liquid to 10000mL, centrifuge, fill and sterilize.

[0020] Ginseng decoction: Ginseng was purchased from Bozhou Huqiao Pharmaceutical Co., Ltd. (supplier to the Pharmacy Department of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine). The ginseng roots were removed, and the ginseng was crushed. 600 g of the crushed ginseng was weighed and mixed with distilled water at a ratio of 1:10, stirred evenly, and soaked overnight. The ginseng extract was extracted at 95 ℃ for 2 h, cooled, and filtered. Then, a second extraction and filtration were performed. The two extracts were mixed and concentrated using a rotary evaporator at 70 ℃ to a concentration of 267 g / L ginseng extract. After concentration, the extract was frozen for later use.

[0021] Red ginseng and astragalus decoction: The Chinese medicinal materials were purchased from Bozhou Huqiao Pharmaceutical Co., Ltd. (supplier of the Pharmacy Department of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine). 500g of red ginseng and 500g of astragalus were decocted in 1000mL of distilled water for 1 hour. The decoction was decocted twice as usual. The two decoctions were then mixed and concentrated to 1g·mL using an R202 rotary evaporator. -1 .

[0022] Experiment Example 3: Comparative Evaluation of the "Heatiness" Induced by Shandong Donkey-hide Gelatin Syrup animal experiments 1. Animal grouping After arrival, the rats were numbered by their ears and housed in cages of four for one week of acclimatization, maintaining a 12h / 12h light / dark cycle (lights off at 8:00 AM and on at 8:00 PM). They had free access to water and food, and the housing was maintained at 21±1℃ and 45% humidity. After the acclimatization period, based on baseline uniformity of weight and open space, the rats were divided into seven groups of 12 each: normal control group; low-dose Shandong Ejiao Syrup group; medium-dose Shandong Ejiao Syrup group; high-dose Shandong Ejiao Syrup group; Dong'e Compound Ejiao Syrup group; ginseng group; and red ginseng and astragalus group.

[0023] 2. Administration After the rats completed their acclimatization period, the medium-dose group of Shandong donkey-hide gelatin oral liquid was administered it by gavage for 28 days. The dosage was calculated based on the human dosage in the drug's instructions, using a conversion factor between human and rat body surface area, and was 40 ml / day / 70 kg body weight. 6.3 = 3.6 ml / kg / d. The low-dose group of Shandong Ejiao Oral Liquid was administered Shandong Ejiao Oral Liquid by gavage for 28 days, with a dosage of 1.8 ml / kg / d. The high-dose group of Shandong Ejiao Oral Liquid was administered Shandong Ejiao Oral Liquid by gavage for 28 days, with a dosage of 7.2 ml / kg / d. The Dong'e Compound Ejiao Oral Liquid group of rats was administered Dong'e Compound Ejiao Oral Liquid by gavage for 28 days, with the dosage calculated based on the human dosage in the drug instructions using a conversion factor between human and rat body surface area, which was 60 ml / d / 70 kg body weight. 6.3 = 5.4 ml / kg / d. The ginseng group received ginseng decoction by gavage for 28 days, with a dosage of 4 g raw ginseng / kg / d according to literature reports. The red ginseng and astragalus group received red ginseng and astragalus decoction by gavage for 28 days, with a dosage of 4 ml / rat / d (divided into two doses of 2 ml / day) according to literature reports. The normal control group received physiological saline by gavage for 28 days, 2 mL / time, once a day.

[0024] 3. Observation indicators 3.1 Observation of the general condition of the animals After the acclimatization period, and 1 week, 2 weeks, 3 weeks and 4 weeks after gavage, the animals in each group were observed in general condition, weighed, and their weekly food and water intake were calculated. Their rectal temperature was measured, and their urine and feces were photographed and recorded. Their shape and color were observed, and their tongues were photographed and recorded.

[0025] 3.2 Open Field Experiment On day 29, rats in each group underwent an open field experiment one hour after gavage. The rats were placed in the center of the open field chamber, and their activities were recorded over 6 minutes using an infrared camera system and video synthesizer. The Xinruan analysis system was used to record the rats' movement trajectories and related behavioral indicators: total distance traveled, distance traveled in the central area, number of times the rats entered the central area, and time spent in the central area. After each rat completed the experiment, the residue in the open field chamber was cleaned, and the scent of the previous rat was removed to avoid affecting the behavior of subsequent animals.

[0026] 3.3 Elevated Cross Maze Experiment One hour after gavage on day 30, rats in each group underwent an elevated cross maze test. The time and number of times rats entered open arms and closed arms were measured using an elevated cross maze apparatus and SuperMaze software. Rats were placed in the central area of ​​the intersection of any open arm, with their heads facing the direction of the open arm. Video recording was used to capture their movement trajectory over 5 minutes, and relevant behavioral indicators were analyzed. The following indicators were recorded: ① Number of times the rat's body part or more entered any open arm (OE); ② Number of times the rat's body part or more entered any closed arm (CE); ③ Time in Open Arm (OT); ④ Time in Close Arm (CT); ⑤ Total distance; ⑥ Distance in Close Arm. OE% and OT% were calculated from ① to ④. The calculation methods are as follows: OE% = OE / (OE+CE)×100%, OT% = OT / (OT+CT)×100%.

[0027] 4. Tissue sampling After the above tests were completed, on day 32 of the experiment, the animals in each group were anesthetized with isoflurane. Blood was collected from all animals using the orbital venous plexus sampling method and stored in centrifuge tubes containing anticoagulant. After standing at room temperature for 30 min, the supernatant was collected by centrifugation at 3500 rpm for 15 min and stored at -80°C for later analysis. Liver tissue was rapidly dissected and separated from the animals on ice. After rinsing off impurities and blood with pre-chilled PBS (0.01M, pH=7.4), the tissue was rapidly cooled with liquid nitrogen and stored at -80°C for later analysis.

[0028] 5. ELISA testing The levels of malondialdehyde (MDA), interleukin-6 (IL-6), and ATP in the plasma of rats in each group were detected using an enzyme-linked immunosorbent assay (ELISA) kit.

[0029] Thaw the frozen serum tissue (-20℃, 4℃), centrifuge (13000 rpm / 10 min), collect the supernatant, and perform the assay according to the relevant kit instructions. Within 5 min of reaction termination, input the concentration values ​​of each standard on the test kit, and measure the optical density (OD) of each group sequentially at a wavelength of 450 nm using a microplate reader. After obtaining the standard curve, substitute the sample OD values ​​into the obtained regression equation to calculate the concentration of each sample. Take the average value of the replicates. If the test sample is diluted, the final sample concentration needs to be multiplied by the dilution factor.

[0030] 6. Experimental Results 6.1 General Observation (1) Feces Figure 1 In the group of rats in the "heatiness" category (red ginseng and astragalus), the feces became drier and harder, with reduced moisture and a darker color after gavage at weeks 2, 3, and 4, indicating that this group of rats exhibited "heatiness"-like feces. No significant differences in fecal morphology were observed in the other groups throughout the experimental period.

[0031] (2) Urine Figure 2 In the group of rats in the red ginseng and astragalus group (indicating internal heat), the urine color turned yellow and cloudy after gavage at weeks 1, 2, 3, and 4. Similarly, the urine color of rats in the normal control group and the high-dose Shandong donkey-hide gelatin oral liquid group also turned yellow and cloudy after gavage at weeks 1, 2, 3, and 4. This suggests that these three groups of rats exhibited urine resembling internal heat. No significant differences in urine color and condition were observed in the remaining groups throughout the experimental period.

[0032] (3) Tongue appearance Figure 3 In the ginseng group, the tongue color of rats changed from red to dark after gavage at weeks 1, 2, 3, and 4, suggesting that the rats in this group exhibited a tongue appearance resembling that of someone experiencing "internal heat." No significant differences in tongue appearance were observed in the other groups throughout the entire experimental period.

[0033] (4) Weight Table 1. Body weight of rats in each group ( ± SE) Compared to the normal control group, rats in the medium-dose Shandong donkey-hide gelatin oral liquid group showed significant weight gain after 1, 2, 3, and 4 weeks of gavage. No significant differences in weight were observed in the other groups throughout the experimental period. This suggests that medium-dose Shandong donkey-hide gelatin intervention may have spleen-strengthening / tonifying effects.

[0034] (5) Rectal temperature Table 2. Results of rectal temperature in each group of rats ( ± SE) No significant differences in rectal temperature were observed among the groups of rats throughout the experimental period.

[0035] 6.2 Behavioral Results (1) Elevated cross maze experiment Table 3 Results of the elevated cruciform maze experiment in each group of rats ( ± SE) In the elevated cross maze test, compared with the normal control group, the percentage of rats entering the open arm in the Dong-A Compound Donkey-hide Gelatin Oral Liquid group and the ginseng group was significantly lower (p<0.05; p<0.05), and the percentage of rats remaining in the open arm in the Dong-A Compound Donkey-hide Gelatin Oral Liquid group and the ginseng group was significantly lower (p<0.05; p<0.05). No significant differences were observed among the other groups. This suggests that these two groups of rats exhibited anxiety ("heat-like") behavior.

[0036] (2) Light and dark box experiment Table 4 Results of the light-dark chamber experiment for each group of rats ( ± SE) In the light-dark chamber experiment, compared with the normal control group, the distance traveled in the light zone was significantly reduced in the compound donkey-hide gelatin oral liquid group and the ginseng group (p<0.01; p<0.01), and the number of times the rats entered the light zone was significantly reduced in the compound donkey-hide gelatin oral liquid group (p<0.05). No significant differences were found among the other groups. This suggests that these two groups of rats exhibited anxiety ("heat-like") behavior.

[0037] 6.3 Results of plasma-related factor testing Table 5. Results of plasma-related factors in rats of each group ( ± SE) In the biochemical detection experiment of plasma-related factors, compared with the normal control group, the plasma MDA level of rats in the Compound Donkey-hide Gelatin Oral Liquid group and the Red Ginseng and Astragalus group was significantly increased (p<0.01; p<0.01), while no significant differences were found among the other groups. Compared with the normal control group, the plasma ATP level of rats in the Compound Donkey-hide Gelatin Oral Liquid group, the Ginseng group, and the Red Ginseng and Astragalus group was significantly increased (p<0.0001; p<0.01; p<0.05), while no significant differences were found among the other groups. Compared with the normal control group, the plasma IL-6 level of rats in the Red Ginseng and Astragalus group was significantly increased (p<0.01), while no significant differences were found among the other groups. This suggests that these three groups of rats exhibited "heatiness"-like biochemical indicators, namely an increase in "dryness factors."

[0038] Based on the above general observations, behavioral tests, and plasma-related factor biochemical test results, the following preliminary conclusions can be drawn: (1) After 4 weeks of oral administration of ginseng or red ginseng and astragalus decoction, rats showed a typical "heatiness" phenotype and an increase in plasma-related factors MDA, ATP and IL-6, suggesting that the positive control was reliable. (2) Among the detection indicators, the general observation combined with the detection of anxiety-like behavior can effectively reflect the "heatiness" appearance of animals. The changes in MDA, ATP and IL-6 in plasma biochemical indicators are more obvious and can be used as "heatiness factors" to evaluate whether the animal is experiencing "heatiness". (3) Based on a comprehensive evaluation of the above indicators, among the tested drugs, the competitor Dong-A E-Jiao Compound E-Jiao Syrup induced rats to exhibit symptoms of internal heat or changes in "dryness factor" in plasma biochemistry, but the other tested drugs did not show significant effects, suggesting that continuous administration of Compound E-Jiao Syrup can induce "internal heat" in animals. (4) After intervention with Shandong Ejiao Oral Liquid (low, medium and high doses), no "heatiness" symptoms or changes in dryness factors were induced in rats. This suggests that Fupai Ejiao Shandong Ejiao Oral Liquid is less likely to cause "heatiness" in animals than competing products such as Dong'e Ejiao Compound Ejiao Oral Liquid and other ginseng-containing decoctions.

[0039] Experiment Example 4: Comparative Evaluation of the Efficacy of Shandong Donkey-hide Gelatin Oral Liquid in Improving Nutritional Anemia 1. Experimental animals: SD strain clean-grade purebred male healthy rats, weighing 50g-60g, provided by Shanghai Xipu Yibikai Experimental Animal Co., Ltd.

[0040] 2. Dosage grouping: Three dosage groups (low, medium, and high doses, i.e., 3.4 mL / kg, 6.7 mL / kg, and 20 mL / kg) and a control group. Because the human intake of the test samples was relatively large, the samples were concentrated three times under vacuum.

[0041] 3. Test methods 3.1 Experimental Phases This experiment is divided into two phases. Phase 1 was the establishment of a rat anemia model. Healthy male SD rats weighing 50g-60g were used to acclimatize to the experimental environment for 5 days and fed a low-iron diet and sterile double-distilled water. Starting from the third week of the experiment, tail blood was collected from a portion of the rats each week to measure hemoglobin levels. When the hemoglobin level of most animals was <100g / L, the body weight and relevant blood cytological indicators of all rats were measured.

[0042] Phase 2 was the recovery period for anemic rats. Anemic rats were selected based on a hemoglobin level <109 g / L. They were randomly divided into four groups according to hemoglobin content: a control group, a low-dose group, a medium-dose group, and a high-dose group, with 10 rats in each group. Rats were administered sterile double-distilled water, low-, medium-, and high-dose Shandong donkey-hide gelatin oral liquid daily via gavage, at doses of 0 mL / kg, 3.4 mL / kg, 6.7 mL / kg, and 20 mL / kg, respectively, for 30 consecutive days. The day after the last gavage, venous blood was collected from the inner canthus of the rat's eye. Hemoglobin content was determined using the methemoglobin method; serum ferritin content was determined using radioimmunoassay; and the free protoporphyrin content in erythrocytes was determined by fluorescence spectrophotometry after erythrocyte destruction with an ethyl acetate-acetic acid mixture and extraction with hydrochloric acid.

[0043] 3.2 Husbandry conditions: Rats were housed in a barrier system environment with a temperature of 18-22℃ and a relative humidity of 40-60%. All rats were housed individually in cages with free access to food and water, which was sterile double-distilled water.

[0044] 3.3 Data Analysis: Analytical variance was performed on the measurement data using SPSS 10.0 for Windows statistical software.

[0045] 4. Experimental Results 4.1 Changes in rat body weight: The body weight of rats in the high- and medium-dose groups of Shandong Ejiao Oral Liquid was significantly higher than that of the control group after the experiment (P<0.05 or P<0.01).

[0046] Table 6. Changes in body weight of rats before and after supplementation with Shandong donkey-hide gelatin oral liquid. 4.2 Changes in hemoglobin content in rats before and after supplementation with Shandong donkey-hide gelatin oral liquid: After supplementation with Shandong donkey-hide gelatin oral liquid, the hemoglobin content in rats in the high, medium and low dose groups was significantly higher than that in the control group (P<0.05).

[0047] Table 7. Changes in hemoglobin levels in rats before and after supplementation with Shandong donkey-hide gelatin oral liquid. 4.3 Serum ferritin and erythrocyte protoporphyrin levels: After rats were supplemented with Shandong donkey-hide gelatin oral liquid, the serum ferritin levels in the high- and medium-dose groups were significantly higher than those in the control group (P<0.01).

[0048] Table 8. Changes in serum ferritin and erythrocyte protoporphyrin in rats before and after supplementation with Shandong donkey-hide gelatin oral liquid. 5. Experimental Conclusions The effect of Shandong Ejiao Oral Liquid on improving nutritional anemia was evaluated using a rat anemia model. After establishing a rat anemia model by feeding them a low-iron diet, Shandong Ejiao Oral Liquid was administered by gavage at doses of 3.4 mL / kg, 6.7 mL / kg, and 20 mL / kg to observe its effect on improving nutritional anemia. The results showed that the hemoglobin content in the high, medium, and low dose groups of Shandong Ejiao Oral Liquid was significantly higher than that in the control group; the serum ferritin content in the high and medium dose groups was significantly higher than that in the control group. This indicates that Shandong Ejiao Oral Liquid has an effect on improving nutritional anemia.

Claims

1. A Shandong donkey-hide gelatin oral liquid composition that does not cause internal heat, characterized in that, It consists of the following ingredients by weight: 50-70g of donkey-hide gelatin, 30-50g of wolfberry, 70-90g of prepared rehmannia root, 15-25g of astragalus root, and 90-110g of jujube.

2. The Shandong donkey-hide gelatin oral composition with non-heat-inducing effect according to claim 1, characterized in that, It consists of the following ingredients by weight: 60g donkey-hide gelatin, 40g wolfberry, 80g prepared rehmannia root, 20g astragalus root, and 100g jujube.

3. A method for preparing a Shandong donkey-hide gelatin oral composition that does not cause internal heat, as described in any one of claims 1-2, characterized in that, Includes the following steps: (1) The ethanol percolation method was used to extract wolfberry, astragalus and jujube, and the percolate was concentrated to obtain percolate concentrate. (2) The prepared Rehmannia glutinosa was extracted by water extraction. The extracted water extract and the percolation concentrate were subjected to cold precipitation. The cold precipitation was filtered and concentrated to obtain the water extract percolation concentrate. (3) After the above-mentioned water-extracted percolate, donkey-hide gelatin liquid and sweetener are prepared once, heated, cold-sedimented and filtered, stabilizer is added, heated a second time, cold-sedimented, the cold-sedimented liquid is centrifuged and filled, and sterilized.

4. The method for preparing the Shandong donkey-hide gelatin oral composition that does not cause internal heat according to claim 3, characterized in that, The percolation process for wolfberry, astragalus, and jujube is as follows: percolation is carried out using 60% ethanol as a solvent; specifically, before percolation, atractylodes macrocephala and wolfberry are soaked in a solvent that is 8 to 10 times the volume of the medicinal materials, and the percolation time is 24 hours.

5. The method for preparing the Shandong donkey-hide gelatin oral composition that does not cause internal heat according to claim 3, characterized in that, The process of water extraction of Rehmannia glutinosa is as follows: add water to Rehmannia glutinosa to decoct and obtain decoction, then filter the decoction to obtain filtrate.

6. The method for preparing the Shandong donkey-hide gelatin oral composition that does not cause internal heat according to claim 5, characterized in that, The process of water extraction of Rehmannia glutinosa is as follows: Rehmannia glutinosa is soaked in 8 times the amount of water for 0.5 hours, heated to 80-85℃, and extracted by low-temperature ultrasonic extraction at 200W and 40kHz for 2 hours; it is decocted at least three times, the decoctions are combined, firstly filtered with gauze to remove suspended impurities, and then purified by passing through three ultrafiltration membranes in sequence: 20000Da for the first ultrafiltration membrane, 10000Da for the second ultrafiltration membrane, and 5000Da for the third ultrafiltration membrane. The permeate from the third ultrafiltration is collected, and 0.1% citric acid is added to adjust the pH to 5.5-6.0 to obtain the decoction of Rehmannia glutinosa.

7. The method for preparing the Shandong donkey-hide gelatin oral composition that does not cause internal heat according to claim 3, characterized in that, The steps are as follows: (1) Soak wolfberry, astragalus and jujube in 0.8 times the amount of 60% ethanol at room temperature for 0.5 hours, then add 60% ethanol to submerge the surface of the medicinal materials. After soaking for 24 hours, percolate. The total amount of 60% ethanol used for soaking, soaking and percolation is 10 times the amount of the medicinal materials. Let the percolate stand for 4 hours, filter it, and concentrate the filtrate under reduced pressure to a relative density of 1.15-1.20 for later use. (2) Soak Rehmannia glutinosa in 8 times the amount of water for 0.5 hours, heat to 80-85℃, and extract with low-temperature ultrasound at 200W and 40kHz for 2 hours. Filter the decoction. Extract the residue twice, adding 4 times and 3 times the amount of water respectively, for 1.5 hours each time. Combine the decoctions. First, use gauze to coarsely filter and remove suspended impurities, then purify it by passing it through three ultrafiltration membranes in sequence: 20000Da for the first ultrafiltration membrane, 10000Da for the second ultrafiltration membrane, and 5000Da for the third ultrafiltration membrane. Collect the permeate from the third ultrafiltration membrane, add 0.1% citric acid to the permeate, and adjust the pH to 5.5-6.0 to obtain the decoction of Rehmannia glutinosa. (3) Add the decoction of Rehmannia glutinosa to the above percolation concentrate, stir for 30 minutes, cool and settle at 2~10℃ for 48 hours, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.05-1.10; (4) Grind the donkey-hide gelatin into powder, weigh it accurately, add 6 times the amount of purified water, stir until completely dissolved, keep it simmering for 30 minutes, filter, and set aside; (5) Add an appropriate amount of white sugar to 4 times the amount of water, boil and dissolve to obtain sugar syrup, and set aside; (6) Mix the concentrated extract of medicinal slices, the enzymatic hydrolysate of donkey-hide gelatin, and the sugar solution, stir for 30 minutes, boil for 30 minutes, cool and settle at 2~8℃ for 48 hours, centrifuge, add water to the centrifuged liquid to the specified amount, centrifuge, fill and sterilize.

8. The application of a Shandong donkey-hide gelatin oral composition that does not cause internal heat in the preparation of health products for improving nutritional anemia.

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