Preparation method and application of ginseng food-medicinal homologous plant as base product

Abstract

The present application provides a preparation method and application of ginseng food homologous plant as a base product, specifically relates to a fresh ginseng and hawthorn extract preparation to increase the content of rare saponins and arginine disglycoside AFG and to be made into tablets and used in the application of drugs for preventing or treating diabetic retinopathy. The present application uses fresh ginseng and hawthorn as raw materials, mixes fresh ginseng and different hawthorn extracts according to the proportion, extracts and retains the natural original saponins in the maximum limit, generates rare saponin component groups such as ginsenoside Rg3, Rk1 and Rg5, and increases small molecule substances such as AFG by enzymatic reaction. The preparation process does not damage the enzyme and protein activity, can effectively enhance the pharmacological activity, fully plays the efficacy of fresh ginseng and hawthorn. The present application can be applied to the field of medicine. It has the effects of enhancing antioxidant capacity, resisting aging, enhancing lipid-lowering and the like.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine compatibility and chemical transformation of traditional Chinese medicine, specifically involving the preparation method and application of products based on ginseng, a plant that is both medicinal and edible. Background Technology

[0002] The theory and dynamics of compatibility in traditional Chinese medicine (TCM) compound prescriptions are a hot topic in TCM theoretical research. The efficacy of TCM compound prescriptions is closely related to the action of their active ingredients or groups of active ingredients. While each herb in a compound prescription exerts its comprehensive pharmacological effect, the co-extraction process involves the full reaction of different substances, which may lead to solubilization and changes in chemical composition, thus producing unique pharmacological activities. Drug pairs refer to the most basic form of compatibility between drugs, forming the fundamental elements of a prescription, and sometimes representing the simplest prescription; their clinical application has been extensively studied. Summary of the Invention

[0003] Ginseng is one of the commonly used traditional Chinese medicines, belonging to the Araliaceae family of the plant Panax ginseng (Panax ginseng). Panax ginseng The dried roots and rhizomes of *C. mey.* (CA Mey.). The main active components of ginseng are ginsenosides, among which the main active components, ginsenosides Rg1, Rb1, and Re, are the content determination indicators for ginseng medicinal materials specified in the 2015 edition of the *Chinese Pharmacopoeia*. According to literature reports, low doses of ginsenosides have a certain protective effect against chronic liver injury, while high doses of ginsenosides can significantly reduce collagen deposition in liver tissue, improve the degree of liver fibrosis, and have an anti-liver fibrosis effect.

[0004] Hawthorn is a plant used both as food and medicine, primarily known for its effects on regulating blood lipids, protecting the liver, lowering blood pressure, and aiding digestion, making it widely applicable. The main active ingredients in hawthorn include flavonoids, flavans and their polymers, organic acids, and triterpenoids. Among these, the flavonoids in hawthorn, such as hyperoside and ursolic acid, have significant cholesterol-lowering and blood lipid-regulating effects. In modern TCM clinical practice, ginseng and hawthorn are often combined to invigorate qi and strengthen the spleen, promote qi circulation and remove blood stasis, and nourish the heart and generate fluids.

[0005] Although ginseng has been proven to have significant hypoglycemic activity, there is still no clear research conclusion on whether the increased content of rare ginsenosides induced by the process and the generation of Maillard reaction products (MRPs) in the hawthorn-ginseng combination system can enhance its hypoglycemic effect, especially its intervention efficacy against diabetic complications (such as retinopathy).

[0006] Currently, ginsenosides undergo changes when ginseng is decocted with some traditional Chinese medicines. For example, in the literature:

[0007] Zhang Shiyang et al. studied the changes in saponin composition before and after combining ginseng with Atractylodes macrocephala and its therapeutic effect on chronic atrophic gastritis. HPLC was used to determine the changes in saponin chemical composition before and after combining ginseng with Atractylodes macrocephala. Treatment of a rat model of chronic atrophic gastritis showed that the combination of ginseng and Atractylodes macrocephala was more effective in treating gastric mucosal damage in rats with chronic atrophic gastritis, possibly related to the increased content of ginsenosides and the formation of new components.

[0008] Du Qinqin et al. used high performance liquid chromatography coupled with electrospray mass spectrometry (HPLC-ESI-MS) to study the decoction, residue and precipitate of ginseng with different proportions of aconite or coptis. A total of 8 kinds of ginsenosides were identified. It was found that the content of ginsenosides in the solution was significantly reduced after the addition of aconite or coptis. Further research proved that some components in aconite or coptis caused the hydrolysis of ginsenosides.

[0009] Zheng Fei et al. used high-performance liquid chromatography-time-of-flight mass spectrometry (HPLC-TOF-MS) to analyze the changes in ginsenosides before and after combining ginseng with hawthorn. The results showed that the contents of ginsenosides Rg1 and Rb1 significantly decreased after combining ginseng with hawthorn, while the contents of ginsenosides Re, Rd, Rg2, Rg3, F2, and Rh1 significantly increased. The combination of ginseng and hawthorn altered the types and contents of ginsenoside components. This study provides material basis data for the changes in components after combining ginseng with hawthorn in clinical prescriptions.

[0010] The above-mentioned existing techniques for combining ginseng with traditional Chinese medicine demonstrate that this combination can alter the types and content of ginsenosides. The existing techniques for combining ginseng with hawthorn mainly have the following problems:

[0011] (1) Limited raw materials: In the existing technology, the research on the compatibility of ginseng and hawthorn all uses dried medicinal slices. However, there are significant differences in the chemical composition spectrum and biological activity between fresh and dried products. Furthermore, the traditional medicinal value of fresh ginseng has not been developed and utilized, which leads to the research on compatibility mechanism deviating from the real material basis.

[0012] (2) The reaction efficiency problem: the traditional water decoction method (100℃ normal pressure) has mild reaction conditions, which makes it difficult to fully activate the conversion pathway of ginsenosides and Maillard reaction, resulting in insufficient generation of target active ingredients and restricting the improvement of efficacy.

[0013] (3) Problems with component recognition: Existing research only focuses on changes in saponin components, but ignores non-saponin active components (such as AFG and furans) produced by Maillard reaction between reducing sugars and amino acids under high temperature conditions, resulting in incomplete analysis of the pharmacodynamic material basis. Summary of the Invention

[0015] To address the aforementioned problems in the existing technology, this invention provides a method for preparing and applying products derived from ginseng, a plant that is both medicinal and edible, and a method for promoting the conversion of rare ginsenosides and the generation of Maillard reaction products when used in combination.

[0016] This invention fully overcomes the shortcomings of existing technologies that combine ginseng and hawthorn. It uses fresh ginseng as the main raw material and combines it with hawthorn extract, which has functions such as regulating oxidation, lowering blood lipids, and reducing blood pressure and cholesterol. The two work together to fully utilize the original effects of fresh ginseng and hawthorn without adding any chemical components. At the same time, it makes full use of the fact that the types and contents of ginsenosides in fresh ginseng increase during the processing, and rare saponin components such as highly active ginsenosides Rg3, Rk1, and Rg5, as well as small molecules such as arginine disaccharide AFG, thus enriching its medicinal value.

[0017] This invention is achieved through the following technical solution:

[0018] The preparation method and application of products derived from ginseng (a plant used in both medicine and food) include the following steps:

[0019] (1) Fresh ginseng and hawthorn were first ground and crushed using a tissue grinder until the diameter was less than 1.0 mm;

[0020] (2) Extract the crushed hawthorn with 0-70% ethanol at 30-70℃ for 1 h to obtain hawthorn extract;

[0021] (3) Add hawthorn extract to fresh ginseng tissue that has been crushed into a paste, with the amount of hawthorn extract being 3 to 6 times the weight of fresh ginseng. Then add purified water in an amount of 20 to 50 times the weight of fresh ginseng.

[0022] (4) The above mixture is placed in a high-pressure steam sterilizer at 100~130°C and reacted for 1~2 h;

[0023] (5) After the reaction is complete, cool to room temperature, filter, concentrate the filtrate under reduced pressure and freeze dry to obtain a product containing rare saponins and AFG.

[0024] Extensive experimental studies have revealed that when fresh ginseng is used in combination with different hawthorn extracts, it can generate rare saponin components such as ginsenosides Rg3, Rk1, and Rg5, as well as components such as arginine disaccharide AFG. This preparation process can effectively enhance pharmacological activity and fully exert the medicinal effects of fresh ginseng and hawthorn.

[0025] As a preferred embodiment of the present invention, the hawthorn extract is an extract obtained by extraction at 50°C and 70% ethanol for 1 hour. 70% ethanol refers to an aqueous solution of ethanol, wherein the volume percentage of ethanol is 70%.

[0026] As a preferred embodiment of the present invention, the amount of hawthorn extract added is 4 times the mass of fresh ginseng.

[0027] As a preferred embodiment of the present invention, the amount of purified water added is 30 times the mass of the fresh ginseng.

[0028] As a preferred embodiment of the present invention: the mixture is placed in a high-pressure steam sterilizer at 120°C and reacted for 2 hours.

[0029] Extensive experimental research has shown that by using the aforementioned extraction process parameters, it is possible to maximize the extraction and retention of natural native saponins while generating rare saponin components such as ginsenosides Rg3, Rk1, and Rg5, as well as components such as arginine disaccharide AFG, thereby enhancing the medicinal value.

[0030] Furthermore, in the tablet preparation method of the present invention, the total tablet content is 400 mg, and its components are as follows: 178 mg of rare saponins and AFG products, 120 mg of maltodextrin, 80 mg of honey powder, 16 mg of povidone (PVP-k30), and 6 mg of magnesium stearate. The above components (except magnesium stearate) are first thoroughly mixed and then passed through an 80-mesh sieve. Finally, magnesium stearate is added and mixed. The tablets are then compressed using the direct powder compression method to produce tablets. The qualified test standards are: hardness 60.7 N, friability 0.69%, and disintegration time 10 min. After the finished product passes the inspection, it is printed and packaged.

[0031] The inventive principles and beneficial effects of this invention include:

[0032] This invention describes a method for promoting the conversion of rare ginseng saponins and the generation of Maillard reaction products by using fresh ginseng and hawthorn extract in combination, and for preparing tablets based on a maltodextrin-honey powder-povidone composite excipient system. The method utilizes fresh ginseng and hawthorn extract in combination, maximizing the preservation of the natural components of fresh ginseng. This method is significantly superior to using ginseng slices and other processed ginseng products. Furthermore, the processing generates rare saponin components not found in fresh ginseng, effectively compensating for the shortcomings of existing technologies for combining ginseng with other ingredients. It maximizes the preservation of natural saponins while generating rare saponin components such as ginsenosides Rg3, Rk1, and Rg5, as well as arginine disaccharide AFG.

[0033] This invention also demonstrates for the first time that the product containing rare saponins and AFG can reduce damage to 661W retinal ganglion cells and RPE retinal pigment epithelial cells induced by a high-glucose microenvironment by activating intracellular antioxidant stress pathways, thereby maintaining the integrity of retinal cell structure and function. Attached Figure Description

[0034] Figure 1Comparative HPLC chromatographic analysis of ginsenoside components in the ginseng-hawthorn combination system: Example 1 is the HPLC chromatogram of ginsenoside components extracted from ginseng alone; Example 20 is the HPLC chromatogram of changes in ginsenoside components extracted from ginseng-hawthorn co-extracted in the prior art; Example 5 is the HPLC chromatogram of ginsenosides under the optimal ratio of ginseng-hawthorn in this patent invention. Detailed Implementation

[0035] Example 1:

[0036] Select 20 parts by weight of fresh ginseng, clean the ginseng, grind and crush it with a tissue pulverizer until the diameter is less than 1.0 mm, add methanol at a ratio of 1:30 by weight, perform ultrasonic extraction twice, centrifuge, combine the supernatants, then concentrate and adjust the volume, and then perform high performance liquid chromatography to detect ginsenoside components and AFG.

[0037] Example 2:

[0038] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with water at 50℃ for 1 h to obtain an aqueous extract. The hawthorn aqueous extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. A certain amount of purified water was then added and mixed thoroughly. Purified water was then added at a ratio of 1:30 based on the weight of fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0039] Example 3:

[0040] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 30% ethanol at 50°C for 1 h to obtain an extract. The hawthorn extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120°C for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0041] Example 4:

[0042] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 50% ethanol at 50°C for 1 h to obtain an extract. The hawthorn extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120°C for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0043] Example 5:

[0044] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50°C for 1 h to obtain an extract. The hawthorn extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120°C for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0045] Example 6:

[0046] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 30°C for 1 h to obtain an extract. The hawthorn extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120°C for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0047] Example 7:

[0048] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 70℃ for 1 h to obtain an extract. The hawthorn extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0049] Example 8:

[0050] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 20 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0051] Example 9:

[0052] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 40 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0053] Example 10:

[0054] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 120 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0055] Example 11:

[0056] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:20 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0057] Example 12:

[0058] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:40 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0059] Example 13:

[0060] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:50 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 120℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0061] Example 14:

[0062] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50°C for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 100°C for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0063] Example 15:

[0064] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 110℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0065] Example 16:

[0066] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 130℃ for 2 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0067] Example 17:

[0068] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 130℃ for 1 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0069] Example 18:

[0070] Ginseng and hawthorn were purified and ground using a tissue grinder to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 130℃ for 1.5 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0071] Example 19:

[0072] Ginseng and hawthorn were purified and ground using a tissue homogenizer to a diameter of less than 1.0 mm. Hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain a 70% ethanol extract. The 70% ethanol extract was added to the pulverized fresh ginseng tissue at a ratio of 20 parts by weight of fresh ginseng and 80 parts by weight of hawthorn extract. Purified water was then added at a ratio of 1:30 based on the weight of the fresh ginseng, and the mixture was thoroughly mixed. The mixture was placed in a beaker and subjected to a high-pressure steam sterilizer at 130℃ for 2.5 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The ginsenosides and AFG were then detected by high-performance liquid chromatography (HPLC).

[0073] Example 20:

[0074] 10 g of ginseng and 20 g of hawthorn were weighed and decocted in water for 1 hour. After filtration, alcohol was added to make the alcohol content reach 75%. The decoction was then filtered and passed through a 0.22 μm microporous membrane to obtain the ginseng-hawthorn decoction. The changes in the content of ginsenosides were then detected by high performance liquid chromatography. (Wang Renguang, Zhang Ruiduo, Shi Wei, et al. Preliminary study on the changes of ginsenoside components after co-extraction of lipid-lowering and liver-protecting granules[J]. Chinese Journal of Traditional and Folk Medicine, 2017, 26(06):25-28.)

[0075] Example 21:

[0076] The present invention also discloses a method for preparing tablets. The physiologically active drug in the tablets of the present invention is a product containing rare saponins and AFG produced under the above-mentioned optimal combination conditions of fresh ginseng and hawthorn. The excipients are: filler (maltodextrin), flavoring agent (honey powder), binder (povidone PVP-k30), and lubricant (magnesium stearate).

[0077] In the above formula, the problem of difficulty in shaping has been solved. The flavoring agent can mask bitterness and improve taste, making it effective for patients with bitter taste in their mouths.

[0078] The tablets contain a total of 400 mg and are composed of the following components: 178 mg of rare saponins and AFG products, 120 mg of maltodextrin, 80 mg of honey powder, 16 mg of povidone (PVP-k30), and 6 mg of magnesium stearate.

[0079] The above ingredients (except magnesium stearate) are first thoroughly mixed and then passed through an 80-mesh sieve. Finally, magnesium stearate is added and mixed. The powder is directly compressed into tablets. The qualified test standards are: hardness 60.7 N, friability 0.69%, and disintegration time 10 min. After the finished product passes the inspection, it is printed and packaged.

[0080] The contents of rare ginsenoside components and small molecule substance AFG in different embodiments were determined as shown in Tables 1-5. (Primary saponins are represented by the total content of Rb1, Rb2, Rc, and Rd saponins; rare saponins are represented by the content of Rg3; and small molecule substances are represented by the content of AFG). Unit: (mg / g)

[0081] Table 1: Comparison of the contents of primary saponins, rare saponins, and small molecule substances after using different extracts of fresh ginseng and hawthorn in combination.

[0082]

[0083] Table 2: Effects of different mass ratios of fresh ginseng and hawthorn 70% ethanol extract on primary saponins, rare saponins, and small molecule substances.

[0084]

[0085] Table 3: Effects of different materials on primary saponins, rare saponins, and small molecule substances after the combined use of fresh ginseng and hawthorn 70% ethanol extract.

[0086]

[0087] Table 4: Effects of different reaction temperatures on the levels of primary saponins, rare saponins, and small molecules after the combined use of fresh ginseng and hawthorn alcohol extract.

[0088]

[0089] Table 5: Effects of different reaction times on primary saponins, rare saponins, and small molecule substances after the combined use of fresh ginseng and hawthorn alcohol extract.

[0090]

[0091] Table 6: Effects of the present invention's patented technology on primary saponins, rare saponins, and small molecule substances after combining ginseng and hawthorn.

[0092]

[0093] Compared with existing technologies, this patented invention can convert primary saponins into rare saponins to the greatest extent. Under the same conditions of ginseng and hawthorn raw materials, taking the production of ginsenoside Rg3 as an example, the method in the literature can produce 0.57 mg / g of Rg3, while the present invention can produce 5.47 mg / g of ginsenoside Rg3, with a yield more than 5 times that of the literature method.

[0094] HPLC analysis of the above examples revealed that fresh ginseng contains primary saponins such as Rg1, Re, Rb1, Rb2, Rc, and Rd. When fresh ginseng is used in combination with hawthorn, and after reacting at a certain temperature for a certain time in a high-pressure steam sterilizer, the content of primary saponins decreased, and rare ginseng saponins such as Rg3, Rk1, and Rg5, as well as small molecule substances such as arginine disaccharide AFG, were produced. The combination of the two effectively enhanced the pharmacological activity and fully exerted the medicinal effects of fresh ginseng and hawthorn.

[0095] Example 22:

[0096] 1. Drug preparation

[0097] The product obtained in Example 5 of this invention (hereinafter referred to as "component") was dissolved in DMEM high-glucose medium to prepare a stock solution with a concentration of 1000 mg / mL, which was then diluted to concentrations of 1, 2, 4, 8, 16, 32, 64, and 128 mg / mL. Glucose was diluted with DMEM high-glucose medium to a 100 mM solution. The entire process was carried out under aseptic conditions. After the drug was dissolved, it was filtered through a 0.22 μM filter membrane before use.

[0098] 2. Cell Culture

[0099] 661W retinal ganglion cells and RPE retinal pigment epithelial cells were obtained from the Bethune School of Basic Medical Sciences, Jilin University, and cultured in a Thermo incubator at 37°C and 5% CO2. The culture medium used was DMEM containing 10% FBS, 1% penicillin, and 1% streptomycin. The conditions for formal experiments were: under a microscope, both 661W and RPE cells showed normal karyotypes, were adherent, and were in the logarithmic growth phase, with a cell density of 80-90%.

[0100] 3. MTT assay for the effect of TFG on cell viability

[0101] 661W and RPE cells were seeded in 96-well plates and treated with glucose and different concentrations of the same components. Then, 20 μL of MTT solution (5 mg / mL) was added to each well, and the plates were incubated at 37°C for 3.5 h. After the formation of deep blue formazan crystals, the supernatant was discarded. 150 μL of DMSO solution was added to each well to dissolve the formazan crystals, and the plates were shaken to mix for 5 min. The absorbance was then measured at 490 nm using a DR-200B microplate reader. Groups treated with only DMEM medium were normalized to 100% viability.

[0102] 4. Antioxidant activity (MDA, SOD, CAT) detection

[0103] 661W cells and RPE cells were seeded in 6-well culture plates and treated with glucose and different concentrations of components. The cell culture medium was then collected and mixed with RIPA lysis buffer (with protease inhibitors and phosphatase inhibitors added in proportion) for protein extraction. The MDA, SOD, and CAT detection kits were tested according to the instructions, and the absorbance values ​​were measured using a DR-200B microplate reader. The antioxidant activity was calculated based on the absorbance values.

[0104] 5. Data Processing

[0105] All data are expressed as mean ± standard deviation (Mean ± SD) established in different experiments, and analyzed using one-way ANOVA and Bonferroni post-hoc tests. Statistical graphs were generated using GraphPad Prism 8.0.2 software (GraphPad Software, Inc., San Diego, USA). p <0.05, p <0.01 or p <0.001 indicates a statistically significant difference.

[0106] 6. Results

[0107] The protective effects of the components against glucose-induced oxidative stress in 661W and RPE cells are shown in Tables 7, 8, 9, and 10. According to the MTT assay, the effects of different concentrations (1, 2, 4, 8, 16, 32, 64, and 128 mg / mL) of the components on the viability of 661W and RPE cells under high glucose treatment or without glucose supplementation were determined. Glucose treatment significantly decreased cell viability (***). p < 0.001), the given component showed good protective activity (### p < 0.001, , # p <0.05%, the effects of different concentrations (1, 2, 4, 8, 16, 32, 64 and 128 mg / mL) of the component on HUVEC cells with and without glucose treatment were detected using an MDA, SOD and CAT kit. The results showed that glucose treatment significantly increased MDA levels in 661W cells and RPE cells (***). p < 0.001), SOD and CAT levels were significantly reduced (*** p < 0.001); and after component intervention, the abnormal changes in the above indicators were significantly improved. Among them, component (64 mg / mL) showed the best protective effect, and could significantly alleviate glucose-induced oxidative stress damage in 661W cells and RPE cells (compared with the glucose treatment group, ### p< 0.001; ## p < 0.01; # p < 0.05). These experimental data indicate that the component can effectively alleviate glucose-induced oxidative stress damage in 661W and RPE cells. Therefore, the component demonstrates a significant advantage in reducing glucose-induced oxidative stress in 661W and RPE cells.

[0108] Table 7. Effects of components on glucose-induced activity of 661W cells (left) and RPE cells (right).

[0109]

[0110] Table 8. Effects of components on glucose-induced oxidative stress markers (MDA) in 661W and RPE cells.

[0111]

[0112] Table 9. Effects of components on glucose-induced oxidative stress markers (SOD) in 661W and RPE cells.

[0113]

[0114] Table 10 Effects of components on glucose-induced oxidative stress markers (CAT) in 661W and RPE cells

[0115]

[0116] The above embodiments are merely descriptions and illustrations of a method for promoting the conversion of rare ginsenosides and the generation of Maillard reaction products after the combined use of fresh ginseng and hawthorn, and are provided as brief examples. However, the present invention is not limited to the above embodiments. Modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the core spirit of the present invention, and the present invention can be implemented in various ways, but all such modifications and substitutions should be within the protection scope of the present invention, and will not be described in detail here.

Claims

1. A method for preparing a ginsenoside-based product containing rare saponins and AFG, characterized in that, Includes the following steps: (1) Fresh ginseng and hawthorn were first ground and crushed using a tissue grinder until the diameter was less than 1.0 mm; (2) The crushed hawthorn was extracted with 70% ethanol at 50℃ for 1 h to obtain hawthorn extract; (3) Add hawthorn extract to fresh ginseng tissue that has been crushed into a paste, with the amount of hawthorn extract being 3 to 6 times the weight of fresh ginseng. Then add purified water in an amount of 20 to 50 times the weight of fresh ginseng. (4) The above mixture is placed in a high-pressure steam sterilizer at 100~130°C and reacted for 1~2 h; (5) After the reaction is complete, cool to room temperature, filter, concentrate the filtrate under reduced pressure and freeze dry to obtain a product containing rare saponins and AFG.

2. The method according to claim 1, characterized in that: In step (3), the mass ratio of hawthorn extract to fresh ginseng is 4:

1.

3. The method according to claim 1, characterized in that: In step (3), the amount of purified water added is 30 times the mass of the fresh ginseng.

4. The method according to claim 1, characterized in that: The reaction conditions in step (4) are 2 hours in a high-pressure steam sterilizer at 120°C.

5. A tablet, characterized in that: Made from a base of rare saponins and AFG products prepared by any one of claims 1 to 4, each tablet contains the following components: 178 mg of rare saponins and AFG products, 120 mg of maltodextrin, 80 mg of honey powder, 16 mg of povidone (PVP-k30), and 6 mg of magnesium stearate.

6. The use of the product containing rare saponins and AFG prepared by the method of any one of claims 1-4 in the preparation of a medicament for the prevention or treatment of diabetic retinopathy.

7. The application according to claim 6, characterized in that: The product containing rare saponins and AFG reduces damage to 661W retinal ganglion cells and RPE retinal pigment epithelial cells induced by a high-glucose microenvironment by activating intracellular antioxidant stress pathways, thus maintaining the integrity of retinal cell structure and function.

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