Traditional Chinese medicine for diabetes osteoporosis and preparation method thereof

Abstract

This invention discloses a traditional Chinese medicine for diabetic osteoporosis and its preparation method. The medicine is composed of Rehmannia glutinosa (processed), Cornus officinalis, Dioscorea opposita, Alisma plantago-aquatica, Poria cocos, Paeonia suffruticosa, Epimedium brevicornu, Angelica sinensis, and Achyranthes bidentata. Achyranthes bidentata is extracted using a eutectic solvent with choline chloride and oxalic acid in a 1:1 weight ratio before being used in the medicine. The preparation method is as follows: Achyranthes bidentata is extracted using a eutectic solvent to obtain an extract; the remaining herbs are decocted in water, and the two extracts are combined. Pharmacological tests show that this invention can significantly improve loose stools caused by the cloying and greasy nature of Rehmannia glutinosa, and the effect is superior to that of traditional decoction. This invention effectively reduces gastrointestinal side effects and is suitable for long-term use by patients with diabetic osteoporosis.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to a modified Liuwei Dihuang formula for osteoporosis and its preparation method. Background Technology

[0002] Diabetic osteoporosis (DOP) is a systemic metabolic bone disease caused by diabetes, characterized by decreased bone mass per unit volume, altered bone microstructure, reduced bone strength, and increased bone fragility, leading to increased susceptibility to fractures. It is a significant complication of diabetes in the skeletal system, severely impacting patients' quality of life and imposing a substantial economic burden on society.

[0003] Traditional Chinese medicine does not record the diseases "diabetes" and "osteoporosis" as specific names. Based on their clinical characteristics and pathogenesis, most physicians categorize them under "Xiao Ke" (wasting and thirsting syndrome), "Xiao Shen" (wasting and kidney deficiency), "Gu Wei" (bone weakness), and "Gu Bi" (bone pain). Currently, mainstream Chinese medicine considers diabetic osteoporosis to be a musculoskeletal disease caused by dysfunction of multiple organs, falling under the category of "Xiao Ke" combined with "Gu Wei". In discussions of Xiao Ke, *Ren Zhai Zhi Zhi Fang Lun* states, "Heat lurks in the lower body, and kidney deficiency affects it, resulting in thin and weak legs and knees, and aching joints..." *Yi Jing* also states, "Symptoms of Xiao Shen include thinness and leg pain; nourishing kidney water is appropriate," etc. These symptom descriptions resemble bone weakness and bone pain. The kidney is key. The kidney is the foundation of innate essence, storing essence, which generates marrow, which nourishes bones; bones are governed by the kidney. When kidney essence is deficient, the source of bone marrow production is insufficient, and bones lack nourishment, leading to withered bones, fragile bones, and bone weakness. Since the liver and kidneys share a common origin, prolonged kidney deficiency inevitably damages the liver. Liver and kidney deficiency leads to separation of bones and muscles, atrophy of tendons and bones, and exacerbation of osteoporosis. Traditional Chinese medicine believes that the basic pathogenesis of osteoporosis is primarily kidney essence deficiency, but it is often accompanied by liver and spleen deficiency, along with other factors such as blood stasis and phlegm dampness. DOP (Diabetic Osteopathy) is mostly seen in patients with long-term diabetes. Therefore, although liver and kidney deficiency is the primary cause, prolonged yin deficiency extends to yang, and the disease penetrates the collaterals. Essentially, it is a disease of deficiency in the root and excess in the branch. Its pathogenesis is not a simple linear cause-and-effect relationship, but rather a complex relationship of multiple deficiencies with blood stasis, deficiency within excess, and yin disease affecting yang. Therefore, treatment mainly focuses on tonifying the liver and kidneys, regulating yin and yang, while also addressing blood stasis and unblocking the collaterals to achieve the therapeutic goal of strengthening tendons and bones.

[0004] The Yin-Yang balance involved in diabetic osteoporosis and the osteoblast-osteoclast coupling share similarities in multiple dimensions. From an origin perspective, although osteoblasts and osteoclasts have distinct functions, both the mesenchymal stem cells from which osteoblasts originate and the hematopoietic stem cells from which osteoclasts originate are derived from the late mesoderm, which aligns with the interdependent relationship of Yin and Yang. Functionally, osteoblasts are responsible for bone formation, while osteoclasts are responsible for bone resorption; their mutual hierarchy and struggle mirror the opposing relationship of Yin and Yang. Theoretically, the *Suwen* (Plain Questions) states, "Yang transforms into Qi, Yin forms into shape." Based on the characteristic of "Yin forming into shape," osteoblasts and their dominant bone production can be attributed to "Yin"; based on the characteristic of "Yang transforming into Qi," osteoclasts and their dominant bone resorption can be attributed to "Yang." Clearly, the Yin-Yang theory of Traditional Chinese Medicine and the bone metabolic balance of the osteoblast-osteoclast coupling exhibit a high degree of consistency across multiple levels. Maintaining skeletal stability requires osteoblast-osteoclast coupling to achieve a dynamic balance between bone formation and bone resorption, and periodic bone remodeling. In clinical cases, an imbalance between these two processes can affect the maintenance of bone metabolic homeostasis.

[0005] Liuwei Dihuang Wan was first recorded in "Xiao'er Yaozheng Zhijue". This formula was modified from Shenqi Wan in Zhang Zhongjing's "Jinkui Yaolue" and is a classic formula for nourishing Yin and tonifying the kidney. The main functions of Liuwei Dihuang Wan are to nourish Yin and tonify the kidney, replenish essence and marrow, and it has a good effect on various symptoms of liver and kidney Yin deficiency.

[0006] In recent years, numerous clinical studies have confirmed that Liuwei Dihuang pills can effectively treat type 2 diabetic osteoporosis, increase bone density, alleviate pain symptoms, and help improve oxidative stress and blood sugar control. For example, in 2023, the Sixth People's Hospital of Heze City published an article titled "Application Analysis of Liuwei Dihuang Pills in Patients with Type 2 Diabetic Osteoporosis" in the 13th issue of *Diabetes New World*, selecting 95 patients with T2 DOP and confirming that Liuwei Dihuang can improve the patients' clinical indicators. In 2022, An Juan et al. published an article titled "Efficacy Evaluation of Liuwei Dihuang Pills in Treating Type 2 Diabetic Osteoporosis" in the 7th issue of *Journal of Hunan University of Traditional Chinese Medicine*, randomly dividing 60 patients with T2 DOP into a treatment group and a control group. After a 3-month observation period, they found that Liuwei Dihuang pills could effectively treat T2 DOP. DOP increases bone density, alleviates pain symptoms, and helps improve oxidative stress and blood sugar control. Ding Wenjun et al. published "The Effect and Mechanism of Liuwei Dihuang Pill on Rats with Diabetic Nephropathy and Osteoporosis" in Volume 58, Issue 1 of the Journal of Nankai University (Natural Science Edition) in 2025, which confirmed that Liuwei Dihuang improves bone metabolism indicators and bone tissue morphology parameters in rats with diabetic nephropathy and osteoporosis by regulating the RANKL / RANK / OPG signaling pathway. Liuwei Dihuang preparations play an important role in the treatment of DOP.

[0007] On the other hand, from the perspective of clinical application safety and compliance, long-term use of Liuwei Dihuang carries a certain risk of gastrointestinal adverse reactions. The processed Rehmannia root in Liuwei Dihuang is sticky and viscous, hindering digestion. Physicians throughout history have pointed out that it "easily promotes dampness and fullness, impeding the digestion and absorption of the spleen and stomach, often causing poor appetite, abdominal distension, and loose stools, especially detrimental to those with spleen deficiency and excessive phlegm." Long-term use of Liuwei Dihuang pills may lead to abdominal distension, loose stools, and loss of appetite. Therefore, those with weak spleen and stomach, loose stools, and indigestion should use Liuwei Dihuang pills with caution. In response, the National Medical Products Administration (NMPA) issued an announcement in February 2021 regarding the revision of the instructions for Liuwei Dihuang preparations, stating that according to NMPA adverse reaction monitoring data, Liuwei Dihuang preparations have reported adverse reactions such as diarrhea, abdominal pain, abdominal distension, nausea, vomiting, and gastrointestinal discomfort.

[0008] Patients with diabetic osteoporosis are mostly elderly, whose spleen and stomach functions already decline with age, leading to reduced digestive and absorptive capacity. In this context, long-term use of Liuwei Dihuang Wan (Six-Ingredient Rehmannia Decoction) may induce or worsen gastrointestinal discomfort such as bloating and loose stools due to the cloying and stomach-irritating properties of prepared Rehmannia root. This not only affects the patient's quality of life but may also lead to treatment interruption, thus impacting the overall efficacy of the treatment for diabetic osteoporosis. Therefore, how to rationally modify Liuwei Dihuang Wan to enhance its therapeutic efficacy for diabetic osteoporosis while effectively reducing or eliminating the loose stools caused by its cloying and stomach-irritating properties, making it more suitable for the long-term chronic disease management of elderly patients with diabetic osteoporosis, is an urgent technical problem to be solved. Summary of the Invention

[0009] Through long-term clinical practice, the inventors discovered that by adding Epimedium, Achyranthes bidentata, and Angelica sinensis to the classic Liuwei Dihuang formula, and especially by employing specific extraction techniques for Achyranthes bidentata and controlling its dosage in the overall formula, this problem can be effectively solved. Epimedium, which enters the liver and kidney meridians, tonifies the kidney yang, benefits essence and qi, and strengthens tendons and bones. Its pungent, sweet, and warm properties, acting as a counteracting agent, also have the effect of seeking yin within yang. This disease is a complication of diabetes, and prolonged illness affects the collaterals; therefore, adding Angelica sinensis nourishes blood and yin, and also invigorates blood circulation and removes blood stasis. Adding Achyranthes bidentata, with its bitter, sweet, sour, and neutral properties, enters the liver and kidney meridians, simultaneously tonifying the liver and kidneys, strengthening tendons and bones, and removing blood stasis and clearing the collaterals. The combined effects of these herbs tonify the liver and kidneys, regulate yin and yang, and also invigorate blood circulation, clear the collaterals, and relieve pain.

[0010] This invention is achieved in the following manner:

[0011] This invention provides a traditional Chinese medicine for diabetic osteoporosis, which, by weight, is composed of the following components:

[0012] The principal and assistant herbs for tonifying the kidneys and nourishing yin are: 20-28 parts of Rehmannia glutinosa, 10-14 parts of Cornus officinalis, and 6-12 parts of Dioscorea opposita.

[0013] Adjuvant herbs for promoting diuresis and eliminating turbidity: Alisma plantago-aquatica 6-12 parts, Poria cocos 6-12 parts, Paeonia suffruticosa 6-12 parts;

[0014] Added medicinal materials: 10-15 parts of Epimedium, 10-15 parts of Angelica sinensis, and 12-20 parts of Achyranthes bidentata. The Achyranthes bidentata is extracted with a eutectic solvent of choline chloride and oxalic acid in a weight ratio of 1:1 before use.

[0015] Furthermore, the dosage of Achyranthes bidentata satisfies the following requirements:

[0016] = Achyranthes bidentata dosage × Achyranthes bidentata extract wet product extraction coefficient × Achyranthes bidentata polysaccharide content in Achyranthes bidentata extract wet product × (1 - Achyranthes bidentata extract wet product drying loss) ÷ Rehmannia glutinosa dosage = 0.01-0.03;

[0017] The extraction coefficient of the wet extract is the weight of the wet extract divided by the weight of the raw Achyranthes bidentata after extraction. The content of Achyranthes bidentata polysaccharides in the dry extract is determined by the phenol-sulfuric acid method. The phenol-sulfuric acid method is based on the "Determination of Polysaccharides in Different Processed Products of Achyranthes bidentata" published by Tianjin Institute of Pharmaceutical Sciences in "Traditional Chinese Medicine". The specific steps include: using glucose as a reference, measuring the absorbance at a wavelength of 490 nm using the phenol-sulfuric acid method, and calculating the content of Achyranthes bidentata polysaccharides in the dry extract using the external standard method.

[0018] Furthermore, the traditional Chinese medicine for diabetic osteoporosis is composed of the following components: 24g Rehmannia glutinosa, 12g Cornus officinalis, 12g Dioscorea opposita, 9g Paeonia suffruticosa, 9g Poria cocos, 9g Alisma plantago-aquatica, 12g Epimedium brevicornu, 12g Angelica sinensis, and 15g Achyranthes bidentata.

[0019] Furthermore, the traditional Chinese medicine for diabetic osteoporosis may also be composed of the following components: 24g of Rehmannia glutinosa, 12g of Cornus officinalis, 12g of Dioscorea opposita, 9g of Paeonia suffruticosa, 9g of Poria cocos, 9g of Alisma plantago-aquatica, 10g of Epimedium brevicornu, 10g of Angelica sinensis, and Achyranthes bidentata; the dosage of Achyranthes bidentata is calculated as: Rehmannia glutinosa dosage × 0.01 ÷ Achyranthes bidentata extract wet product extraction coefficient ÷ Achyranthes bidentata extract wet product dry product polysaccharide content ÷ (1 - Achyranthes bidentata extract wet product drying loss).

[0020] Furthermore, the traditional Chinese medicine for diabetic osteoporosis may also be composed of the following components: 24g of Rehmannia glutinosa, 12g of Cornus officinalis, 12g of Dioscorea opposita, 9g of Paeonia suffruticosa, 9g of Poria cocos, 9g of Alisma plantago-aquatica, 15g of Epimedium brevicornu, 15g of Angelica sinensis, and Achyranthes bidentata; the dosage of Achyranthes bidentata is calculated as: Rehmannia glutinosa dosage × 0.03 ÷ Achyranthes bidentata extract wet product extraction coefficient ÷ Achyranthes bidentata extract wet product dry product polysaccharide content ÷ (1 - Achyranthes bidentata extract wet product drying loss).

[0021] A second aspect of the present invention provides a method for preparing the traditional Chinese medicine for diabetic osteoporosis, comprising the following steps:

[0022] S1. Achyranthes bidentata is extracted using a eutectic solvent with a choline chloride to oxalic acid weight ratio of 1:1 and a water content of 10%-30%.

[0023] S2. Prepared Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis are decocted in water twice to obtain the decoction.

[0024] S3. Add the Achyranthes bidentata extract from S1 to the decoction from S2, stir well, and the product is obtained.

[0025] Furthermore, in the method for preparing the traditional Chinese medicine for diabetic osteoporosis, the eutectic solvent extraction of Achyranthes bidentata comprises the following steps:

[0026] S1. The eutectic solvent is composed of choline chloride and oxalic acid in a weight ratio of 1:1 and a water content of 10%-30% by weight. The dried Achyranthes bidentata is pulverized and passed through a 40-mesh sieve and added to the eutectic solvent. The solid-liquid weight ratio is 1:5-10. The mixture is stirred at 25-50℃ for 1-3 hours, filtered, and the filtrate is collected.

[0027] S2. Add anhydrous ethanol to the filtrate until the alcohol content is 70%-85%, let it stand at 1-15℃ for 8-24 hours, centrifuge to collect the precipitate, and obtain the wet product of Achyranthes bidentata extract.

[0028] The present invention provides a novel, green process for extracting Achyranthes bidentata using a eutectic solvent extraction (DES) method. Compared to traditional water extraction, DES extraction offers milder conditions and lower energy consumption, resulting in higher extraction rates and purity of the extracted plant polysaccharides, along with better bioactivity and integrity. In addition to polysaccharides, Achyranthes bidentata contains a variety of active ingredients, including sterones and triterpenoid saponins, making its composition complex and extraction challenging.

[0029] This invention application provides further optimized process parameters, including a preferred eutectic solvent composition (choline chloride to oxalic acid weight ratio 1:1, water content 10%-30%) and corresponding solid-liquid ratio, extraction temperature, alcohol precipitation conditions, and other process parameters. It was unexpectedly found that the Achyranthes bidentata polysaccharide extracted by this process is significantly better than traditional methods in regulating intestinal flora activity, and can thus more effectively alleviate the loose stool side effect caused by the greasy and cloying nature of Rehmannia glutinosa.

[0030] Furthermore, the method for preparing the traditional Chinese medicine for diabetic osteoporosis includes the following steps: decocting Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis in water.

[0031] S1. Grind Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis into powder and pass through a 40-mesh sieve. Mix them together and add 10-20 times the total weight of the dried herbs in water. Soak for 30-60 minutes. First, bring to a boil over high heat, then simmer over low heat for 30-60 minutes. Filter and collect the first decoction.

[0032] S2. Add the dregs to 5-10 times the amount of water as dried herbs, simmer over low heat for 30-60 minutes, filter, and collect the second decoction filtrate.

[0033] S3. Combine the first decoction filtrate and the second decoction filtrate, mix well, and obtain the decoction liquid.

[0034] A third aspect of the present invention provides an Achyranthes bidentata extract suitable for use in traditional Chinese medicine compositions. This Achyranthes bidentata extract is prepared by: extracting Achyranthes bidentata using a eutectic solvent system of choline chloride and oxalic acid to obtain a wet product, drying it, determining the content of Achyranthes bidentata polysaccharides in the dry product and the loss on drying, and then storing it in a sealed container.

[0035] Preferably, the Achyranthes bidentata extract provided by this invention is a stable intermediate product with uniform and controllable quality, and can be prepared in large quantities and stored in sealed containers. For primary healthcare institutions or patients who do not have the conditions for eutectic solvent extraction, this intermediate product can be used directly in combination with the decoction of the other eight medicinal materials, which lowers the application threshold and has good practicality and promotional value.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] 1. Reasonable composition, enhancing efficacy and reducing side effects

[0038] This invention is based on the classic formula of Liuwei Dihuang Decoction, with the addition of Epimedium (to seek Yin within Yang), Angelica sinensis (to nourish blood, promote blood circulation, and unblock collaterals), and Achyranthes bidentata (to tonify the liver and kidneys, strengthen tendons and bones, and remove blood stasis and unblock collaterals). The combined effects of these herbs are to tonify the liver and kidneys, regulate Yin and Yang, promote blood circulation, unblock collaterals, and relieve pain. It is especially suitable for the pathogenesis of long-term osteoporosis in diabetic patients.

[0039] 2. The Achyranthes bidentata extraction process reduces gastrointestinal side effects.

[0040] A low-melting-point solvent system of choline chloride-oxalic acid was used to extract Achyranthes bidentata polysaccharides. This method offers mild extraction conditions, low energy consumption, and high extraction rate and purity, while better preserving the natural structure and bioactivity of the polysaccharides. Unexpectedly, it was found that the Achyranthes bidentata polysaccharides extracted using this process were significantly superior to traditional water extraction in regulating intestinal flora activity, thus effectively alleviating the side effect of loose stools caused by the cloying and irritating properties of Rehmannia glutinosa, and improving patient compliance with long-term use.

[0041] 3. Achyranthes bidentata is fed according to the content of dry matter markers to reduce the quality variation of the drug.

[0042] In this invention, the dosage of Achyranthes bidentata is calculated based on the dry content of the extract, effectively correcting for quality fluctuations caused by natural differences in the key medicinal material Achyranthes bidentata from different origins and batches. This calculation method ensures that the contribution of Achyranthes bidentata's effective components in the final compound formula remains constant, solving the technical problem of traditional Chinese medicine's direct dosage based on "experience-based estimation" and large batch-to-batch quality variations. Furthermore, this extract can be prepared in large quantities, stored in sealed containers, and used immediately. For primary healthcare institutions or patients lacking eutectic solvent extraction capabilities, it can be directly purchased and used, lowering the application threshold. Attached Figure Description

[0043] Figure 1 Example 3: Flowchart of the preparation of Achyranthes bidentata extract (dried product)

[0044] Figure 2 Example 1: Ultraviolet spectrum of Achyranthes bidentata extract (wet product) tested using the phenol-sulfuric acid method. Detailed Implementation

[0045] The following description illustrates exemplary embodiments of the present invention, including various details to aid understanding, and should be considered merely exemplary. Therefore, those skilled in the art will recognize that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the invention. Similarly, for clarity and brevity, descriptions of well-known functions, operations, and structures are omitted in the following description.

[0046] Unless otherwise defined, the technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. While similar or identical methods and materials may be applied in experimental or practical applications, the present invention describes materials and methods hereinafter. In case of conflict, the definitions included herein shall prevail.

[0047] Example 1: Preparation of Traditional Chinese Medicine for Diabetic Osteoporosis

[0048] prescription:

[0049] The Achyranthes bidentata used in this embodiment is produced in Jiaozuo City, Henan Province, and is one of the "Four Great Huai Medicines". It is of high quality, but the market supply is unstable.

[0050] Processing:

[0051] Achyranthes bidentata extract:

[0052] S1, Eutectic Solvent: Composed of choline chloride and oxalic acid in a weight ratio of 1:1, with a water content of 10%-30% by weight; specifically, it consists of 1000g choline chloride, 1000g oxalic acid, and 400g water.

[0053] Achyranthes bidentata: Remove impurities, wash, soak thoroughly, remove residual rootlets, cut into sections, dry, and pulverize through a 40-mesh sieve.

[0054] The pulverized Achyranthes bidentata was added to a eutectic solvent with a solid-liquid weight ratio of 1:8 (specifically, 150g of Achyranthes bidentata and 1200g of eutectic solvent); the mixture was stirred at 40℃ for 2 hours, filtered, and the filtrate was collected.

[0055] S2. Add anhydrous ethanol to the filtrate to control the alcohol content to 80%, let it stand at 10°C for 12 hours, centrifuge to collect the precipitate, and obtain the wet product of Achyranthes bidentata extract.

[0056] The remaining 8 ingredients are decocted in water:

[0057] S1. Grind the prepared Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis into powder and pass it through a 40-mesh sieve. Mix them together and add 15 times the total weight of the dried herbs to water (specifically, 900g of herbs to 13500g of water). Soak for 45 minutes, bring to a boil over high heat, then simmer over low heat for 50 minutes. Filter and collect the first decoction.

[0058] S2. Add the dregs to 6 times the amount of water as dried herbs (specifically 5400g of water), simmer over low heat for 50 minutes, filter, and collect the second decoction filtrate.

[0059] S3. Combine the first decoction filtrate and the second decoction filtrate, mix well, and obtain the decoction liquid.

[0060] merge:

[0061] Add all the wet extract of Achyranthes bidentata to the decoction of the remaining 8 ingredients, stir well, and you will get a traditional Chinese medicine for diabetic osteoporosis.

[0062] Analysis of wet product of Achyranthes bidentata extract:

[0063] Wet weight of Achyranthes bidentata extract:

[0064] After extraction, the sample was accurately weighed and a total of 61g was obtained. The wet extract ratio of Achyranthes bidentata extract is 61g / 150g = 0.41.

[0065] Loss on drying of wet Achyranthes bidentata extract:

[0066] 29%.

[0067] Content of dried Achyranthes bidentata polysaccharides in wet Achyranthes bidentata extract:

[0068] 18.9%.

[0069] Method for determining the loss on drying of Achyranthes bidentata extract (wet product):

[0070] The determination of loss on drying was performed according to the method described in Section 0831 of the 2025 edition of the Chinese Pharmacopoeia, using the reduced pressure method. Specific parameters included isothermal drying at 60°C under reduced pressure, and drying to constant weight at a pressure below 2.67 kPa.

[0071] Loss on drying (%) = (Initial wet weight - Weight after drying) / Initial wet weight × 100%.

[0072] Method for determining the content of Achyranthes bidentata polysaccharides in dried Achyranthes bidentata extract (wet product):

[0073] The determination of polysaccharides in different processed forms of Achyranthes bidentata was mainly based on the findings of Lin Xiaohui et al. from Tianjin Institute of Pharmaceutical Research, published in the 8th issue of *Traditional Chinese Medicine* in 2008. The method employed was ultraviolet-visible spectrophotometry. Specifically:

[0074] Preparation of reference solution

[0075] Take an appropriate amount of D-anhydrous glucose reference standard, accurately weigh it, and add water to prepare a solution containing 0.04 mg per ml.

[0076] Preparation of standard curve

[0077] Accurately measure 0.2 ml, 0.4 ml, 0.6 ml, 0.8 ml, and 1.0 ml of the reference solution into separate 10 ml stoppered test tubes. Add distilled water to each tube to a final volume of 2.0 ml, shake well, add 1.0 ml of 5% phenol solution to each tube, mix well, and then quickly add 5.0 ml of concentrated sulfuric acid. Shake for 5 minutes, heat in a boiling water bath for 15 minutes, remove, and immediately cool in a cold water bath for 30 minutes. Measure the absorbance at 490 nm using ultraviolet-visible spectrophotometry (General Chapter 0401, Chinese Pharmacopoeia 2025). Plot a standard curve with absorbance on the ordinate and concentration on the abscissa.

[0078] Determination method

[0079] 1. Preparation of the test sample

[0080] Take an appropriate amount of Achyranthes bidentata extract, mix it evenly, and spread it evenly in a pre-dried flat weighing bottle to constant weight, with a thickness not exceeding 5 mm. Dry it in a 60℃ vacuum drying oven to constant weight. Take about 0.1 g of this dried sample, weigh it accurately, place it in a 100 ml volumetric flask, add water, sonicate to dissolve, and dilute to the mark. Shake well and set aside.

[0081] 2. Content determination

[0082] Accurately measure 1.0 ml of the above test solution and place it in a 10 ml stoppered test tube. Following the method under the preparation of the standard curve, starting from "adding 1.0 ml of 5% phenol solution", determine the absorbance according to the method. Read the amount equivalent to anhydrous glucose from the standard curve and calculate to obtain the result.

[0083] Example 2: Preparation of wet Achyranthes bidentata extract

[0084] prescription:

[0085] 5000g of Achyranthes bidentata

[0086] The Achyranthes bidentata used in this embodiment is grown in Shandong Province, and its quality is slightly lower than that of the authentic product from Shandong. However, the supply is sufficient and stable.

[0087] Achyranthes bidentata extract:

[0088] S1, Eutectic Solvent: Composed of choline chloride and oxalic acid in a weight ratio of 1:1, with a water content of 20% by weight.

[0089] Achyranthes bidentata: Remove impurities, wash, soak thoroughly, remove residual rootlets, cut into sections, dry, and pulverize through a 40-mesh sieve.

[0090] The crushed Achyranthes bidentata was added to a eutectic solvent with a solid-liquid weight ratio of 1:6; the mixture was stirred at 45°C for 1.5 hours, filtered, and the filtrate was collected.

[0091] S2. Add anhydrous ethanol to the filtrate to control the alcohol content to 75%, let it stand at 10°C for 24 hours, centrifuge to collect the precipitate, and obtain the wet product of Achyranthes bidentata extract.

[0092] Analysis of wet product of Achyranthes bidentata extract:

[0093] Wet weight of Achyranthes bidentata extract:

[0094] After extraction, the sample was accurately weighed, yielding a total of 1520g. The wet extraction coefficient of the Achyranthes bidentata extract was 1520g / 5000g = 0.304.

[0095] Loss on drying of wet Achyranthes bidentata extract:

[0096] 35%.

[0097] Content of dried Achyranthes bidentata polysaccharides in wet Achyranthes bidentata extract:

[0098] 15.5%.

[0099] Note: The content determination in this step is the dry product content, that is, the wet product of Achyranthes bidentata extract is dried and then accurately weighed for determination. This is because the drying loss of the wet product is relatively high and fluctuates greatly in practice. This method of determining and labeling the dry product content can eliminate the drawbacks of the wet product content determination and labeling.

[0100] Example 3 Preparation of dried Achyranthes bidentata extract

[0101] Raw materials:

[0102] Example 2: 1000g of wet Achyranthes bidentata extract

[0103] Preparation process:

[0104] The wet Achyranthes bidentata extract was spread evenly on a stainless steel tray and placed in a vacuum oven for drying at 60°C for 12 hours. The product was then pulverized and passed through an 80-mesh sieve, collected, and sealed for storage.

[0105] Weight of dried Achyranthes bidentata extract obtained:

[0106] 619g.

[0107] Loss on drying of dried Achyranthes bidentata extract:

[0108] 1.5%.

[0109] Content of Achyranthes bidentata polysaccharides in dried Achyranthes bidentata extract:

[0110] 15.3%.

[0111] Loss on drying and content of Achyranthes bidentata polysaccharide in dried product were both determined using the method described in Example 1.

[0112] Examples 4-5: Preparation of Traditional Chinese Medicine with Different Achyranthes bidentata Amounts

[0113] prescription:

[0114] The traditional Chinese medicine prescriptions in this example are shown in the table below;

[0115] In Example 4, the Achyranthes bidentata used was from Shandong Province, which is the same batch of medicinal material as in Example 2: Achyranthes bidentata added = 240g × 0.01 ÷ Achyranthes bidentata extract wet product extraction coefficient (i.e.: 1520g / 5000g = 0.304) ÷ 15.5% ÷ (1 - 35%) = 78g.

[0116] In Example 5, the Achyranthes bidentata used was from Shandong Province, which is the same batch of medicinal material as in Example 2: Achyranthes bidentata added = 240g × 0.03 ÷ Achyranthes bidentata extract wet product extraction coefficient (i.e.: 1520g / 5000g = 0.304) ÷ 15.5% ÷ (1 - 35%) = 234g.

[0117] Processing:

[0118] Same as in Example 1.

[0119] Example 6: Preparation of Traditional Chinese Medicine by Directly Adding Dried Achyranthes bidentata Extract

[0120] prescription:

[0121] In this embodiment, Achyranthes bidentata is directly added to the dried Achyranthes bidentata extract prepared in Example 3.

[0122] Based on the 200g weight of the medicinal materials in the prescription, and the extraction coefficient of the wet Achyranthes bidentata extract and the drying loss of the wet Achyranthes bidentata extract in this batch of medicinal materials (see Example 2 for details), it can be determined that the required dry Achyranthes bidentata extract is 200g × 0.304 × (1 - drying loss of the wet Achyranthes bidentata extract) = 39.5g.

[0123] Processing:

[0124] Achyranthes bidentata extract:

[0125] Take 39.5g of the dried Achyranthes bidentata extract prepared in Example 3 directly.

[0126] The remaining 8 ingredients are decocted in water:

[0127] The preparation process is the same as that of the other 8 ingredients in Example 1, prepared by decoction.

[0128] merge:

[0129] Add 39.5g of Achyranthes bidentata extract to the decoction of the remaining 8 ingredients and stir well to obtain a traditional Chinese medicine for diabetic osteoporosis.

[0130] Comparative Example 1: Preparation of Traditional Chinese Medicine by Decoction

[0131] prescription:

[0132] Same as in Example 1.

[0133] Processing:

[0134] Decoct all nine Chinese medicinal herbs in the prescription in water:

[0135] S1. Grind the prepared Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, Angelica sinensis, and Achyranthes bidentata into powder and pass it through a 40-mesh sieve. Mix them together, add water at 15 times the total weight of the dried herbs, soak for 45 minutes, bring to a boil over high heat, then simmer over low heat for 50 minutes. Filter and collect the first decoction.

[0136] S2. Add the dregs to 6 times the amount of water as dried herbs, simmer over low heat for 50 minutes, filter, and collect the second decoction filtrate.

[0137] S3. Combine the first decoction filtrate and the second decoction filtrate, mix well, and obtain the decoction liquid.

[0138] Comparative Example 2: Preparation of Traditional Chinese Medicine Drugs Using Achyranthes bidentata Extract Purified with Water and Alcohol

[0139] The Achyranthes bidentata herb was extracted according to the method disclosed in patent CN 113350386 B.

[0140] prescription:

[0141] Same as in Example 1.

[0142] Processing:

[0143] Achyranthes bidentata extract:

[0144] S1. Take Achyranthes bidentata, add distilled water and extract at 70℃ for 50 min, then boil and extract for 1.5 h, filter to obtain water extract and residue;

[0145] S2. The residue is first extracted in a 40% (w / w) ethanol aqueous solution for 1.5 hours to obtain a primary extract and a filter residue. The filter residue is then extracted in a 70% (w / w) ethanol aqueous solution for 1.5 hours to obtain a secondary extract. The primary and secondary extracts are then combined to obtain an alcohol extract.

[0146] S3. After combining the aqueous extract and the alcohol extract, the extract was subjected to saturation equilibrium by adsorption using AB-8 macroporous resin, and then eluted with 50% (w / w) ethanol aqueous solution at a elution rate of 1 BV / h and a volume of 5 BV of eluent to obtain the Achyranthes bidentata extract.

[0147] The remaining 8 ingredients are decocted in water:

[0148] Same as in Example 1.

[0149] merge:

[0150] Same as in Example 1.

[0151] Comparative Example 3: Preparation of dried product of Achyranthes bidentata extract purified by water and alcohol

[0152] S1. Preparation of wet extract

[0153] The extraction of Achyranthes bidentata was consistent with that in Comparative Example 2.

[0154] S2. Preparation of dried extract

[0155] The solution in S1 is used as an eluent. It is placed in a rotary evaporator with a water bath temperature of 50-60℃, a vacuum degree of -0.08 to -0.1MPa, and a rotation speed of 80-120rpm. It is concentrated until there is no flowing liquid and the bottle wall has a solid wet residue. The residue is then scraped off.

[0156] The wet Achyranthes bidentata extract was spread evenly on a stainless steel tray and placed in a vacuum oven for drying at 60°C for 12 hours. The product was then pulverized and passed through an 80-mesh sieve, collected, and sealed for storage.

[0157] Loss on drying of Achyranthes bidentata extract (dried):

[0158] 1.2%.

[0159] Content of Achyranthes bidentata polysaccharides in dried Achyranthes bidentata extract:

[0160] 10.3%.

[0161] Loss on drying and content of Achyranthes bidentata polysaccharide in dried product were both determined using the method described in Example 1.

[0162] Comparative Example 4: Preparation of Liuwei Dihuang Decoction

[0163] prescription:

[0164] Processing:

[0165] S1. Grind the prepared Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis into powder and pass it through a 40-mesh sieve. Mix them together and add 15 times the total weight of the dried herbs to water (specifically, 660g of herbs to 9900g of water). Soak for 45 minutes, bring to a boil over high heat, then simmer over low heat for 50 minutes. Filter and collect the first decoction.

[0166] S2. Add the dregs to 6 times the amount of water (3960g of water) of the dried herbs, simmer over low heat for 50 minutes, filter, and collect the second decoction filtrate.

[0167] S3. Combine the first decoction filtrate and the second decoction filtrate, mix well, and obtain the decoction liquid.

[0168] Experimental Example 1: Stability Test of Dried Achyranthes bidentata Extracts from Different Extraction Methods

[0169] In accordance with the guidelines for drug stability testing in the Chinese Pharmacopoeia, the dried Achyranthes bidentata extracts prepared in Example 3 and Comparative Example 3 were sealed and placed under accelerated conditions for testing at 40°C and 75% relative humidity for 6 months.

[0170] The properties were observed directly by visual inspection, and the methods for determining the loss on drying and the content of Achyranthes bidentata polysaccharide in dried product were as described in Example 1.

[0171] The test results are as follows:

[0172] in conclusion:

[0173] The dried Achyranthes bidentata extract prepared in Example 3 of this invention (extracted using a eutectic solvent of choline chloride and oxalic acid in a weight ratio of 1:1) remained stable (off-white powder) after being stored for 6 months under accelerated conditions (temperature 40°C, relative humidity 75%), with a drying loss of only 0.3% (from 1.5% to 1.8%). The retention rate of the marker Achyranthes bidentata polysaccharide in the dried product reached 94.1% (from 15.3% to 14.4%). Its stability is significantly better than that of the conventional water extraction and alcohol precipitation method in Comparative Example 3.

[0174] Therefore, the method for preparing Achyranthes bidentata extract described in this invention not only improves the extraction rate of Achyranthes bidentata, but also endows the extract with excellent chemical stability, providing quality assurance for its long-term storage and formulation development as a raw material for traditional Chinese medicine.

[0175] Experimental Example 2: Pharmacodynamic Study of Rats with Improved Stool Flow

[0176] By examining the effects of the traditional Chinese medicine of this invention on a rat model of spleen deficiency and diarrhea, we can verify its efficacy in improving the traditional Chinese medicine symptom of "loose stools".

[0177] 1. Test materials

[0178] Animals: 75 male SPF-grade SD rats, weighing 180-220g, were purchased from Shandong Pengyue Experimental Animal Technology Co., Ltd., Experimental Animal Production License No.: SCXK (Lu) 2023-0002. The animals were acclimatized for 7 days in an environment with a temperature of 22±2℃, relative humidity of 50%-60%, and 12h light-dark alternation before the start of the experiment.

[0179] Rhubarb: Commercially available, identified by a licensed traditional Chinese medicine practitioner as the dried rhizome of Rheum palmatum L.

[0180] 2. Establishment of the diarrhea model

[0181] Preparation of rhubarb decoction:

[0182] Take 200g of rhubarb slices, add 2000mL of water, soak for 30 minutes, then decoct for 30 minutes and filter. Add 1000mL of water to the dregs and decoct for another 20 minutes. Combine the filtrates and concentrate to the equivalent of 1g / mL of raw rhubarb.

[0183] Modeling method:

[0184] Except for the blank control group, rats in the other groups were administered rhubarb decoction (10 mL / kg, equivalent to 10 g / kg of raw drug) by gavage once a day for 14 consecutive days. The successful model was defined as the appearance of obvious loose stools, decreased appetite, lethargy, reduced activity, and perianal soiling.

[0185] The prepared Rehmannia root in Liuwei Dihuang Wan is sticky and greasy, and long-term use can cause gastrointestinal side effects such as diarrhea and abdominal distension. Studies have shown that the "greasy and greasy" properties of prepared Rehmannia root can only be manifested in a state of spleen deficiency. Therefore, this experiment used a rhubarb spleen deficiency diarrhea model. Rats were given rhubarb decoction by gavage to establish a basic state of spleen deficiency. Under this state, they were divided into groups and then treated with the modified Liuwei Dihuang Wan to make the greasy and greasy properties of prepared Rehmannia root manifest, thus constructing an animal model of "prepared Rehmannia root causing loose stools on the basis of spleen deficiency".

[0186] 3. Grouping and Dosing

[0187] In Example 1 and Comparative Examples 1 and 4, the prepared drugs were diluted with purified water to a uniform concentration using the six raw medicinal herbs (Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, and Alisma plantago-aquatica). The gavage volume for each group was 10 ml / kg. The drugs were administered once daily for 14 consecutive days. Gavage administration of rhubarb decoction was discontinued during the treatment period.

[0188] 4. Observation of preparation and measurement methods

[0189] 4.1 General Condition Observation

[0190] The mental state, activity, coat color, perianal cleanliness, and fecal characteristics of rats in each group were observed and recorded daily.

[0191] 4.2 Determination of loose stool rate (%)

[0192] On days 7 and 14 after drug administration, feces were collected from rats in each group within 4 hours after drug administration, and the number of loose stools and the total number of stools were recorded. Fecal characteristics were graded as follows:

[0193] Grading trait description

[0194] Grade 1 normal stool: formed, firm, and dry on the surface.

[0195] Grade 2 soft stool: formed, soft, and moist.

[0196] Grade 3 loose stool: unformed, pasty, or semi-solid.

[0197] Grade 4 watery stool: liquid, amorphous

[0198] Grades 3-4 are included in the number of loose stools. Loose stool rate (%) = Number of loose stools / Total number of stools × 100%.

[0199] 4.3 Determination of fecal moisture content (%)

[0200] On day 14 of drug administration, fresh feces were collected from rats in each group, and the wet weight (W1) was accurately measured. The feces were then dried in a 60℃ oven until constant weight (approximately 24 hours), and the dry weight (W2) was measured. Fecal moisture content (%) = (W1—W2) / W1×100%.

[0201] 4.4 Diarrhea Index

[0202] Diarrhea Index = Loose Stool Rate × Average Loose Stool Grade (Average Loose Stool Grade is calculated based on stool characteristics grading).

[0203] 5. Data Processing and Statistical Analysis

[0204] Data analysis was performed using SPSS 22.0 statistical software. Experimental data are expressed as mean ± standard deviation (x̅ ± s). One-way ANOVA was used for comparisons among multiple groups, and LSD-t test was used for pairwise comparisons between groups. A p-value < 0.05 was considered statistically significant.

[0205] 6. Experimental Results

[0206] Comparison of loose stool rate and fecal water content among rats in different groups (x̅ ± s, n=15)

[0207] Note: Compared with the blank control group, **P<0.01; compared with the model control group, ##P<0.01.

[0208] 7. Conclusion

[0209] The traditional Chinese medicine composition of this invention (Achyranthes bidentata extracted with a eutectic solvent of choline chloride and oxalic acid in a weight ratio of 1:1) can significantly improve diarrhea symptoms in rats with diarrhea model. Compared with the model control group, the rats in the invention group had significantly lower rates of loose stools and lower fecal water content (P<0.01), and the efficacy was significantly better than that of Comparative Example 1 (Achyranthes bidentata raw material decocted with other medicinal materials).

[0210] Further experimental results showed that the proportion of loose stools and stool water content in control group 2 (the original Liuwei Dihuang Decoction formula, excluding Epimedium, Angelica sinensis, and Achyranthes bidentata) were significantly higher than those in the model control group (P<0.01). This indicates that, on the basis of spleen deficiency, the nourishing and cloying properties of Rehmannia glutinosa further aggravated diarrhea symptoms. However, this invention, by adding Epimedium, Angelica sinensis, and Achyranthes bidentata to the Liuwei Dihuang Decoction formula, not only did not aggravate diarrhea but also significantly improved loose stool symptoms and reduced the rate of loose stools. The optimal effect was achieved when Achyranthes bidentata was extracted using a eutectic solvent before being used in the medicine.

[0211] The results indicate that: 1. Adding Epimedium, Angelica sinensis, and Achyranthes bidentata can effectively counteract the cloying and stomach-damaging side effects of Rehmannia glutinosa; 2. Using Achyranthes bidentata extracted with a eutectic solvent of choline chloride and oxalic acid in a weight ratio of 1:1 can more effectively retain or enrich the active ingredients in Achyranthes bidentata that regulate intestinal function, and can significantly enhance the effect of the compound preparation in improving the loose stools caused by the cloying and stomach-damaging effects of Rehmannia glutinosa in Liuwei Dihuang.

[0212] This invention provides a safer and more compliant option for long-term use of Liuwei Dihuang preparations by diabetic osteoporosis patients.

Claims

1. A traditional Chinese medicine for diabetic osteoporosis, characterized in that, It consists of the following components in parts by weight: The principal and assistant herbs for tonifying the kidneys and nourishing yin are: 20-28 parts of Rehmannia glutinosa, 10-14 parts of Cornus officinalis, and 6-12 parts of Dioscorea opposita. Adjuvant herbs for promoting diuresis and eliminating turbidity: Alisma plantago-aquatica 6-12 parts, Poria cocos 6-12 parts, Paeonia suffruticosa 6-12 parts; Added medicinal materials: 10-15 parts of Epimedium, 10-15 parts of Angelica sinensis, and 12-20 parts of Achyranthes bidentata. The Achyranthes bidentata is extracted with a eutectic solvent of choline chloride and oxalic acid in a weight ratio of 1:1 before use.

2. The traditional Chinese medicine for diabetic osteoporosis according to claim 1, characterized in that, The dosage of Achyranthes bidentata is required to meet the following conditions: = Achyranthes bidentata dosage × Achyranthes bidentata extract wet product extraction coefficient × Achyranthes bidentata polysaccharide content in Achyranthes bidentata extract wet product × (1 - Achyranthes bidentata extract wet product drying loss) ÷ Rehmannia glutinosa dosage = 0.01-0.03; The extraction coefficient of the wet Achyranthes bidentata extract is the weight of the wet extract of the batch of Achyranthes bidentata after extraction / the weight of the raw Achyranthes bidentata; the content of Achyranthes bidentata polysaccharides in the wet extract of Achyranthes bidentata is determined by the phenol-sulfuric acid method.

3. The traditional Chinese medicine for diabetic osteoporosis according to claim 1, characterized in that, It is composed of the following ingredients: 24g Rehmannia glutinosa, 12g Cornus officinalis, 12g Dioscorea opposita, 9g Paeonia suffruticosa, 9g Poria cocos, 9g Alisma plantago-aquatica, 12g Epimedium brevicornu, 12g Angelica sinensis, and 15g Achyranthes bidentata.

4. The traditional Chinese medicine for diabetic osteoporosis according to claim 1, characterized in that, It is composed of the following components: 24g of Rehmannia glutinosa, 12g of Cornus officinalis, 12g of Dioscorea opposita, 9g of Paeonia suffruticosa, 9g of Poria cocos, 9g of Alisma plantago-aquatica, 10g of Epimedium brevicornu, 10g of Angelica sinensis, and Achyranthes bidentata; the dosage of Achyranthes bidentata is calculated as: Rehmannia glutinosa dosage × 0.01 ÷ Achyranthes bidentata extract wet product extraction coefficient ÷ Achyranthes bidentata extract wet product dry product content of Achyranthes bidentata polysaccharide ÷ (1 - Achyranthes bidentata extract wet product drying loss).

5. The traditional Chinese medicine for diabetic osteoporosis according to claim 1, characterized in that, It is composed of the following components: 24g of Rehmannia glutinosa, 12g of Cornus officinalis, 12g of Dioscorea opposita, 9g of Paeonia suffruticosa, 9g of Poria cocos, 9g of Alisma plantago-aquatica, 15g of Epimedium brevicornu, 15g of Angelica sinensis, and Achyranthes bidentata; the dosage of Achyranthes bidentata is calculated as: Rehmannia glutinosa dosage × 0.03 ÷ Achyranthes bidentata extract wet product extraction coefficient ÷ Achyranthes bidentata extract wet product dry product polysaccharide content ÷ (1 - Achyranthes bidentata extract wet product drying loss).

6. A method for preparing a traditional Chinese medicine for diabetic osteoporosis as described in claim 1, characterized in that, The steps are as follows: S1. Achyranthes bidentata is extracted using a eutectic solvent with a choline chloride to oxalic acid weight ratio of 1:1 and a water content of 10%-30%. S2. Prepared Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis are decocted in water twice to obtain the decoction. S3. Add the Achyranthes bidentata extract from S1 to the decoction from S2, stir well, and you have obtained the product.

7. The method for preparing a traditional Chinese medicine for diabetic osteoporosis according to claim 6, characterized in that, The eutectic solvent extraction of Achyranthes bidentata comprises the following steps: S1. The eutectic solvent is composed of choline chloride and oxalic acid in a weight ratio of 1:1 and a water content of 10%-30% by weight. The dried Achyranthes bidentata is pulverized and passed through a 40-mesh sieve and added to the eutectic solvent. The solid-liquid weight ratio is 1:5-10. The mixture is stirred at 25-50℃ for 1-3 hours, filtered, and the filtrate is collected. S2. Add anhydrous ethanol to the filtrate until the alcohol content is 70%-85%, let it stand at 1-15℃ for 8-24 hours, centrifuge to collect the precipitate, and obtain the wet product of Achyranthes bidentata extract.

8. The method for preparing a traditional Chinese medicine for diabetic osteoporosis according to claim 6, characterized in that, The decoction of Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis is carried out in the following steps: S1. Grind Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Paeonia suffruticosa, Poria cocos, Alisma plantago-aquatica, Epimedium brevicornu, and Angelica sinensis into powder and pass through a 40-mesh sieve. Mix them together and add 10-20 times the total weight of the dried herbs in water. Soak for 30-60 minutes. First, bring to a boil over high heat, then simmer over low heat for 30-60 minutes. Filter and collect the first decoction. S2. Add the dregs to 5-10 times the amount of water as dried herbs, simmer over low heat for 30-60 minutes, filter, and collect the second decoction filtrate. S3. Combine the first decoction filtrate and the second decoction filtrate, mix well, and obtain the decoction liquid.

9. The method for preparing the traditional Chinese medicine for diabetic osteoporosis according to claim 7, characterized in that, The eutectic solvent extraction step of Achyranthes bidentata further includes: drying the obtained wet Achyranthes bidentata extract, determining the content of Achyranthes bidentata polysaccharides in the dried product and the loss on drying, and storing it in a sealed container.

10. An extract of Achyranthes bidentata obtained by eutectic solvent extraction as described in claim 9, for use in traditional Chinese medicine compositions.

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