Preparation method of Gushukang capsule

Abstract

The application provides a preparation method of Gushukang capsules, and the preparation method comprises the following steps: in the process of preparing Gushukang capsules, a non-D101 type resin column is used to adsorb raw material extract liquid for preparing Gushukang capsules, and then an eluent is used for elution, wherein the ratio between the volume of the raw material extract liquid and the mass of the resin in the resin column is 8-12:1. In the process of preparing Gushukang capsules, the adsorption resin and various parameters are explored, and the yield of phenolic acid, flavone and saponin components in Gushukang capsules is effectively improved.

Description

Technical Field

[0001] This invention relates to the technical field of traditional Chinese medicine preparation, and more specifically to a method for preparing bone-strengthening capsules. Background Technology

[0002] Bone-Soothing Capsules are a marketed traditional Chinese medicine composed of Epimedium, Rehmannia glutinosa, Drynaria fortunei, Astragalus membranaceus, Salvia miltiorrhiza, Auricularia auricula-judae, and Cucumber seeds. It has the effects of tonifying the kidneys and replenishing qi, promoting blood circulation and strengthening bones. It is used for primary osteoporosis caused by kidney deficiency and insufficient qi and blood, with symptoms such as lower back pain, weakness in the lower back and knees, weakness in the lower limbs, difficulty walking, fatigue, dizziness, a reddish or pale tongue, and a flat or weak pulse.

[0003] The 2020 edition of the Chinese Pharmacopoeia discloses the preparation method of Gusukang Capsules:

[0004] Epimedium 1251g, Rehmannia glutinosa 1656g, Drynaria fortunei 828g, Astragalus membranaceus 1251g, Salvia miltiorrhiza 828g, Auricularia auricula-judae 663g, Cucumber seeds 663g

[0005] Preparation: The above seven ingredients are prepared by crushing cucumber seeds, decocting them with wood ear fungus twice with water (2.5 hours the first time and 1.5 hours the second time), filtering, combining the filtrates, and concentrating under reduced pressure to a clear extract with a relative density of approximately 1.05–1.10 (65℃); danshen is extracted twice with ethanol under reflux (2 hours the first time and 1.5 hours the second time), filtering, combining the filtrates, recovering the ethanol, and concentrating to a clear extract with a relative density of approximately 1.20–1.25 (65℃); the danshen residue is decocted three times with the remaining four ingredients, including epimedium, for 1.5 hours the first time and 1 hour each for the second and third times; the decoctions are combined, allowed to stand for 8 hours, and the supernatant is taken and passed through a D101 macroporous adsorption resin, eluted with 70% ethanol, recovering the ethanol from the eluent, and concentrating to a clear extract with a relative density of approximately 1.15–1.20 (65℃); the above clear extract is mixed, dried, pulverized, and encapsulated to make 1000 capsules.

[0006] In the current technology for purifying the decoction of Gushukang capsules using resin adsorption, D101 macroporous adsorption resin is used. Furthermore, the purification process only investigated one representative component, icariin, without considering other components such as tanshinone IIA, salvianolic acid B, naringin, total flavonoids from *Drynaria fortunei*, and astragaloside A. As is well known, the properties of phenolic acids, flavonoids, and saponins in Gushukang capsules vary, and other effective components require further improvement and development. Summary of the Invention

[0007] To address the shortcomings of existing technologies and solve the aforementioned problems, a method for preparing bone-strengthening capsules is proposed, and the following technical solution is provided:

[0008] A method for preparing bone-strengthening capsules involves using a non-D101 type resin column to adsorb the raw material extract for preparing bone-strengthening capsules, followed by elution with an eluent. The ratio between the volume of the raw material extract and the mass of the resin in the resin column is 8-12:1.

[0009] Furthermore, the non-D101 type resin column is an NKA-9 type resin column, an AB-8 type resin column, or an HPD722 type resin column.

[0010] Furthermore, the non-D101 type resin column is an NKA-9 type resin column, and the flow rate of the raw material extract in the resin column is 2-5 mL / min.

[0011] Furthermore, the non-D101 type resin column is an NKA-9 type resin column, the flow rate of the eluent in the resin column is 2.5-10 mL / min, the concentration of the eluent is 70-90%, and the amount of eluent used is 4-10 BV.

[0012] Furthermore, the eluent is ethanol.

[0013] Furthermore, the non-D101 type resin column is an NKA-9 type resin column. The raw material extract for preparing the bone-strengthening capsules is adsorbed using the NKA-9 type resin column. The ratio between the volume of the raw material extract and the mass of the resin in the NKA-9 type resin column is 10:1. The flow rate of the raw material extract in the resin column is 5 mL / min, the flow rate of the eluent in the resin column is 9 mL / min, the concentration of the eluent is 80%, and the amount of eluent used is 6 BV.

[0014] Furthermore, the non-D101 type resin column is an HPD722 type resin column.

[0015] Furthermore, the flow rate of the raw material extract in the resin column is 1-4 mL / min, the flow rate of the eluent in the resin column is 1-8 mL / min, the concentration of the eluent is 80%, and the amount of the eluent is 6-12 BV.

[0016] Furthermore, the ratio between the volume of the raw material extract and the mass of the resin in the NKA-9 resin column is 12:1, the flow rate of the raw material extract in the resin column is 3 mL / min, the flow rate of the eluent in the resin column is 6 mL / min, the concentration of the eluent is 80%, and the amount of the eluent used is 8 BV.

[0017] Due to the adoption of the above technical solutions, the beneficial technical effects of the present invention are as follows:

[0018] 1. This invention explored the adsorption resin and various parameters in the preparation process of bone-strengthening capsules, which effectively improved the yield of phenolic acids, flavonoids and saponins in bone-strengthening capsules;

[0019] 2. This invention also explored the optimal yield values ​​of phenolic acids, flavonoids and saponins, and finally obtained stable and feasible process parameters. Furthermore, regression equations for the influence levels of each process parameter were obtained based on the process parameters. Detailed Implementation

[0020] To enable those skilled in the art to better understand the technical solutions of the present invention, the technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Based on the embodiments in this application, other similar embodiments obtained by those skilled in the art without creative effort should all fall within the scope of protection of this application.

[0021] The dosage and formula of Gushukang Capsules disclosed in the 2020 edition of the Chinese Pharmacopoeia are: Epimedium 1251g, Rehmannia glutinosa 1656g, Drynaria fortunei 828g, Astragalus membranaceus 1251g, Salvia miltiorrhiza 828g, Auricularia auricula-judae 663g, Cucumber seeds 663g. This application studies the above dosage and formula.

[0022] Materials and Instruments

[0023] Macroporous resins: AB-8, ADS-7, D101, D4020, HPD100, NKA-9, X-5, and XAD-2 were purchased from Elest (Shanghai) New Material Technology Co., Ltd.; D301, DM130, HPD300, and HPD722 were purchased from Cangzhou Baoen Adsorption Material Technology Co., Ltd.; LS-46D and LSA-21 were purchased from Shanghai Yuanye Biotechnology Co., Ltd. Standards such as tanshinone IIA, salvianolic acid B, icariin, naringin, total flavonoids from *Drynaria fortunei*, and astragaloside A were purchased from the China National Institutes for Food and Drug Control. Methanol and acetonitrile were chromatographic alcohols (Tedia Reagents, USA); water was double-distilled water; and other reagents were domestically produced analytical grade. The decoction of bone-strengthening capsules was provided by Liaoning Wohua Kangchen Pharmaceutical Co., Ltd. (prepared according to the method of Example 1 of patent CN202111627999.9). Agilent 1200 high performance liquid chromatograph (Agilent Technologies); SHZ-B water bath constant temperature shaker (Shanghai Yuejin Medical Instrument Co., Ltd.); KQ-250E CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); Mettler ME204E electronic balance (Mettler-Toledo Instruments).

[0024] (1) Screening the types of resin columns.

[0025] Weigh 5g each of the pretreated AB-8, ADS-7, D101, D301, D4020, DM130, HPD100, HPD300, HPD722, LS-46D, LSA-21, NKA-9, X-5, and XAD-2 resins and place them in stoppered conical flasks. Add 50mL of bone-strengthening capsule decoction to each flask and shake at 28℃ and 150r / min for 24h in a constant-temperature shaker. Collect the supernatant and determine the content of each component in the supernatant according to HPLC (C1). Wash the saturated resin twice with ultrapure water, then add 50mL of 70% ethanol solution to desorb the resin. Continue shaking and eluting in a constant-temperature shaker for 24h and collect the supernatant to determine the content of each component (C2). Calculate the static adsorption rate and desorption rate according to the following formulas.

[0026] Adsorption rate = (C0-C1) / C0×100%;

[0027] Desorption rate = C2 / (C0-C1)×100%

[0028] C0 represents the content of each component in the decoction of osteoporosis-relieving capsules before adsorption.

[0029] The test results are shown in Table 1 below.

[0030] Table 1. Adsorption rates of six components in the decoction of Shukang capsules by different resin columns.

[0031]

[0032]

[0033] As shown in Table 1 above, there are significant differences in the adsorption rates of the 14 resins for the 6 components contained in the bone susceptibility capsules. AB-8, HPD722, LSA-21, and NKA-9 have higher adsorption rates for phenolic acid components, while AB-8, D301, HPD100, HPD300, HPD722, and NKA-9 have higher adsorption rates for saponin components. NKA-9 has the highest adsorption rate for flavonoid components. After comprehensive consideration, AB-8, HPD722, and NKA-9 macroporous resins were selected to replace the currently used D101 resin. Among them, NKA-9 macroporous resin was the most preferred for purification process research.

[0034] (2) Screening the ratio between the volume of the raw material extract and the mass of the resin in the resin column:

[0035] Accurately weigh 5.0 g of pretreated NKA-9 resin and place it in a stoppered conical flask. Add the decoction of bone-strengthening capsules at precise drug-to-resin (volume) ratios of 6:1, 8:1, 10:1, 12:1, and 16:1, respectively. Shake at 28℃ and 150 r / min for 24 h in a constant-temperature shaker. Collect the supernatant and determine the content of each component in the supernatant using HPLC (C1). Wash the saturated resin twice with ultrapure water, then add 50 mL of 70% ethanol solution to desorb the resin. Continue shaking and eluting in a constant-temperature shaker for 24 h. Collect the supernatant and determine the content of each component (C2). Calculate the adsorption rate of the six components.

[0036] A program was written using Matlab software to calculate the weights according to the entropy weight method. The adsorption rates of the six components were assigned corresponding weight coefficients (W). In the evaluation problem of j evaluation indicators and i evaluated schemes, the entropy weight method was used to calculate the weight coefficient (Wj) of the j-th indicator.

[0037] (1) The weight of the index value of the i-th object under the j-th index (Yij):

[0038]

[0039] (2) The entropy value of the j-th index:

[0040]

[0041] Where, k = 1 / Lnm

[0042] (3)Wj:

[0043]

[0044] The composite index (S) is calculated using the following formula:

[0045]

[0046] Aj represents the adsorption / desorption rate of the six components, and Wj represents the weighting coefficient of the six components.

[0047] The S values ​​corresponding to different loading amounts were calculated to be 82.44%, 87.52%, 89.26%, 87.63%, and 80.59%, respectively. Therefore, the optimal loading amount is considered to be 10:1.

[0048] (3) Screening the flow rate of the raw material extract in the resin column:

[0049] Accurately weigh 5.0 g of pretreated NKA-9 resin and place it in a stoppered conical flask. Load the decoction of bone-soothing capsules into the flask at a ratio of 10:1. Set the loading flow rate to 2.5, 5, 10, 15, and 20 mL / min, respectively. Shake the flask at 28℃ and 150 r / min for 24 h in a constant temperature shaker. Collect the supernatant and determine the content of each component in the supernatant (C1) by HPLC. The saturated resin was washed twice with ultrapure water, and then 50 mL of 70% ethanol solution was added to desorb the resin. The resin was then eluted by shaking in a constant temperature shaker for 24 h. The content of each component (C2) in the supernatant was measured, and the absorption rate and W of the six components were calculated. The corresponding S values ​​were calculated to be 88.46%, 86.17%, 80.22%, 74.81%, and 63.15%, respectively. Considering that the difference between 2.5 mL / min and 5 mL / min is small, the optimal loading flow rate of 5 mL / min was selected from the perspective of improving production efficiency.

[0050] (4) Screening the concentration of the eluent:

[0051] Accurately weigh 5.0 g of pretreated NKA-9 resin and place it in a stoppered conical flask. Load the decoction of bone-strengthening capsules into the flask at a ratio of 10:1 and a flow rate of 5 mL / min. Shake at 28℃ and 150 r / min for 24 h in a constant-temperature shaker. Collect the supernatant and determine the content of each component in the supernatant using HPLC (C1). After complete adsorption, wash with 4 BV purified water at a flow rate of 1.0 mL / min to remove impurities. Elute with 30%, 50%, 70%, 80%, and 90% ethanol (v / v), and continue eluting in a constant-temperature shaker for 24 h. Collect the supernatant and determine the content of each component (C2). The desorption rates and W values ​​of the six components were calculated, and the corresponding S values ​​were 9.27%, 47.25%, 72.24%, 80.42%, and 82.25%, respectively. Considering that the difference between 80% and 90% ethanol is small, 80% was selected as the optimal eluent concentration from the perspective of cost reduction.

[0052] (5) Screening the amount of eluent to be used:

[0053] Accurately weigh 5.0 g of pretreated NKA-9 resin and place it in a stoppered conical flask. Load the decoction of bone-soothing capsules into the flask at a ratio of 10:1 and a flow rate of 5 mL / min. Shake at 28℃ and 150 r / min for 24 h in a constant-temperature shaker. Collect the supernatant and determine the content of each component in the supernatant using HPLC (C1). After complete adsorption, wash with 4 BV purified water at a flow rate of 1.0 mL / min to remove impurities. Elute with 80% ethanol at doses of 2, 4, 6, 8, and 10 BV, respectively. Continue elution in a constant-temperature shaker for 24 h, and collect the supernatant to determine the content of each component (C2). The desorption rates and W values ​​of the six components were calculated, and the corresponding S values ​​were 57.35%, 70.29%, 76.42%, 78.75%, and 82.22%, respectively. After the eluent dosage reached 6 BV, the elution effect changed little. From the perspective of cost reduction, 6 BV was selected as the optimal eluent dosage.

[0054] (6) Screening the flow rate of the eluent:

[0055] Accurately weigh 5.0 g of pretreated NKA-9 resin and place it in a stoppered conical flask. Load the decoction of bone-soothing capsules into the flask at a ratio of 10:1 and a flow rate of 5 mL / min. Shake at 28℃ and 150 r / min for 24 h in a constant-temperature shaker. Collect the supernatant and determine the content of each component in the supernatant using HPLC (C1). After complete adsorption, wash with 4 BV purified water at a flow rate of 1.0 mL / min to remove impurities. Elute with 80% ethanol at 6 BV, with elution flow rates of 2.5, 5, 10, 15, and 20 mL / min, respectively. Continue shaking and eluting in a constant-temperature shaker for 24 h, and then collect and determine the content of each component in the supernatant (C2). The desorption rates and W values ​​of the six components were calculated, and the corresponding S values ​​were 82.36%, 79.52%, 77.26%, 68.24%, and 61.47%, respectively. When the elution flow rate was less than 10 mL / min, the elution effect changed little. From the perspective of improving production efficiency, 10 mL / min was selected as the optimal elution flow rate.

[0056] (7) Response surface methodology:

[0057] Based on the single-factor experiments, the feed rate of the raw material extract, the flow rate of the raw material extract, the concentration of the eluent, the amount of eluent used, and the elution flow rate were selected as five independent variables. +1, 0, and -1 represent the high, medium, and low levels of the independent variables, respectively. The S-value corresponding to the adsorption rate of the six components was used as the response value. The five-factor, three-level response surface methodology was designed using the Box-Behnken central composite experiment principle in Design-expert10 software as follows:

[0058] Table 2 Factor Levels and Coding Table

[0059]

[0060]

[0061] The experimental results are shown in Table 3 below:

[0062] Table 3 Results of the Five-Factor, Three-Level Response Surface Experiment

[0063]

[0064]

[0065] The equations obtained by regression fitting of each factor are as follows:

[0066] S=103.59+0.19A-0.22B-0.34C-0.13D+1.05E+0.17AB-0.94AC+0.071AD-0.44AE+0.063BC-0.15BD+0.073BE-0.29CD-0.73CE+0.24DE-0.95A 2 +0.38B 2 -0.81C 2 -1.53D 2 +0.62E 2 .

[0067] The optimal process conditions for this experiment were obtained by using Design-expert10 software to optimize and predict the levels of each factor: sample loading ratio of 8.3:1, sample loading flow rate of 3.7 mL / min, eluent concentration of 76.25%, eluent volume of 6.2 BV, and elution flow rate of 8.7 mL / min. The maximum predicted S value was 89.75%.

[0068] Three parallel experiments were conducted under the above process conditions, and the average S value of the six components was 88.21%, which deviated from the predicted S value of 89.75% by 1.75%, indicating that the optimized process is stable and feasible.

[0069] The interaction between various parameters was explored using HPD722 resin columns.

[0070] (1) Accurately weigh 5.0 g of pretreated HPD722 resin and place it in a stoppered conical flask. Add the decoction of bone-soothing capsules at precise ratios of 6:1, 8:1, 10:1, 12:1, and 16:1 (drug-to-resin ratio). Shake at 28°C and 150 r / min for 24 h in a constant-temperature shaker. Collect the supernatant and determine the content of each component in the supernatant by HPLC (C1). Wash the saturated resin twice with ultrapure water, then add 50 mL of 70% ethanol solution to desorb the resin. Continue shaking and eluting in a constant-temperature shaker for 24 h. Collect the supernatant and determine the content of each component (C2). Calculate the adsorption rate of the six components. A program was written using Matlab software to calculate the weights according to the entropy weight method. The adsorption rates of the six components were assigned corresponding weight coefficients (W). In the evaluation problem of j evaluation indicators and i evaluated schemes, the entropy weight method was used to calculate the weight coefficient (Wj) of the j-th indicator.

[0071] (1) The weight of the index value of the i-th object under the j-th index (Yij):

[0072]

[0073] (2) The entropy value of the j-th index:

[0074]

[0075] Where, k = 1 / Lnm

[0076] (3)Wj:

[0077]

[0078] The composite index (S) is calculated using the following formula:

[0079]

[0080] Aj represents the adsorption / desorption rate of the six components, and Wj represents the weighting coefficient of the six components.

[0081] The S values ​​corresponding to different loading amounts were calculated to be 80.45%, 84.18%, 87.62%, 89.03%, and 85.69%, respectively. Therefore, the optimal loading amount is considered to be 12:1.

[0082] (2) Accurately weigh 5.0g of pretreated HPD722 resin and place it in a stoppered conical flask. Load the decoction of bone suspensa capsules at a ratio of 12:1. Set the loading flow rate to 1, 3, 5, 8 and 10 mL / min respectively. Shake at 28℃ and 150r / min for 24h in a constant temperature shaker. Extract the supernatant and determine the content of each component in the supernatant (C1) according to HPLC. The saturated resin was washed twice with ultrapure water, and then 50 mL of 70% ethanol solution was added to desorb the resin. The resin was then eluted by shaking in a constant temperature shaker for 24 h. The content of each component (C2) in the supernatant was measured, and the absorption rate and W of the six components were calculated. The corresponding S values ​​were calculated to be 87.61%, 86.54%, 83.82%, 79.88%, and 74.81%, respectively. The S values ​​did not change much when the loading flow rate was set to 1 and 3 mL / min, respectively. When the loading flow rate was 5 mL / min, the S value decreased to 83.82%. The optimal loading flow rate was 3 mL / min.

[0083] (3) Accurately weigh 5.0 g of pretreated HPD722 resin and place it in a stoppered conical flask. Load the decoction of bone-soothing capsules into the flask at a ratio of 10:1 and a flow rate of 5 mL / min. Shake the flask at 28℃ and 150 r / min for 24 h in a constant temperature shaker. Collect the supernatant and determine the content of each component in the supernatant by HPLC (C1). After complete adsorption, wash the flask with 4 BV purified water at a flow rate of 1.0 mL / min to remove impurities. Elute with 80% ethanol at volumes of 2, 4, 6, 8, and 10 BV, respectively. Continue shaking and eluting in a constant temperature shaker for 24 h. Collect the supernatant and determine the content of each component (C2). The desorption rates and W values ​​of the six components were calculated, and the corresponding S values ​​were 52.56%, 60.27%, 66.01%, 76.71%, and 78.34%, respectively. After the eluent dosage reached 8 BV, the elution effect changed little. From the perspective of cost reduction, 8 BV was selected as the optimal eluent dosage.

[0084] Example 1

[0085] The dosage formula of Gushukang Capsules published in the 2020 edition of the Chinese Pharmacopoeia is as follows: Epimedium 1251g, Rehmannia glutinosa 1656g, Drynaria fortunei 828g, Astragalus membranaceus 1251g, Salvia miltiorrhiza 828g, Auricularia auricula-judae 663g, Cucumber seeds 663g.

[0086] Step 1) Take the decoction of Gushukang capsules (the residue after tanshinone alcohol extraction, along with epimedium, rehmannia glutinosa, drynaria fortunei, and astragalus membranaceus, and extract with water 3 times. The first time, add 6 times the amount of water and extract for 1.5 hours. The second time, add 4 times the amount of water and extract for 1 hour. The third time, add 4 times the amount of water and extract for 1 hour. Combine the extracts, let stand, and take the supernatant. Adsorb the decoction through an NKA-9 resin column from top to bottom. Control the flow rate of the decoction at 4 mL / min. Discharge the effluent directly.

[0087] Step 2) After the drug adsorption is completed, each column is washed with 500L of water from top to bottom, and the washing solution is discharged directly.

[0088] Step 3) Use 75% ethanol at 6 times the volume of resin as desorbent for each column, control the flow rate at 9 mL / min, and desorb from top to bottom for 20 minutes. Then start collecting the desorbed liquid, recover the ethanol, and concentrate it to a clear paste with a relative density of about 1.18 (65℃). Dry the clear paste, pulverize it, and fill it into capsules.

[0089] Comparative Example 1

[0090] The bone-strengthening capsules were prepared according to Example 1 of patent CN202111627999.9.

[0091] The content of effective ingredients in the capsules prepared in Example 1 and Comparative Example 1 is compared in Table 4 below.

[0092] Table 4. Content of effective ingredients in capsules prepared in Example 1 and Comparative Example 1

[0093]

[0094] As can be seen from Table 4, the content of all 6 components in Example 1 is greater than that in Comparative Example 1.

[0095] The capsules prepared in Example 1 and Comparative Example 1 were subjected to animal experiments. The specific experimental procedures are shown below.

[0096] Forty 6-month-old specific pathogen-free (SPF) female SD rats, weighing 350-400g, were selected. Husbandry conditions: temperature 24℃, relative humidity 45%. Lighting 12h / d, standard diet, and free access to water and food.

[0097] The capsules of Example 1 and the capsules prepared in Comparative Example 1 were fully dissolved in physiological saline and administered by gavage. Forty-eight rats were acclimatized for two weeks and then randomly divided into four groups: blank control group, model group, Bone-Soothing Kang A group, and Bone-Soothing Kang B group, with 10 rats in each group.

[0098] An osteoporosis model was established. Rats in the model group, Bone-Soothing Kang A group, and Bone-Soothing Kang B group were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (40 mg / kg) and underwent bilateral oophorectomy to establish a postmenopausal osteoporosis model. In the blank control group, some periovarian adipose tissue was removed through the same incision.

[0099] Four weeks after bilateral oophorectomy, the Bone Soothing A group and the Bone Soothing ...

[0100] After administration, all rats were euthanized under anesthesia, and their right femur and fourth lumbar vertebra were removed. The muscles, tendons, and soft tissues attached to the femur and vertebrae were removed. The right femur and lumbar vertebra of the rats were scanned using a dual-energy X-ray absorptiometry (DXA) instrument, and the bone mineral density was measured using measurement software.

[0101] All results were processed and analyzed using SPSS 16.0 statistical software. The t-test was used to compare the sample means between the two groups. The results are expressed as -x±s, and P<0.05 was considered statistically significant.

[0102] The femoral bone mineral density and the bone mineral density of the fourth lumbar vertebra (-x±s, n=10) of each group of rats are shown in Table 5 below.

[0103] Table 5. Results of femoral bone mineral density and fourth lumbar vertebral bone mineral density levels in each group of rats.

[0104] Blank control group 0.492±0.077 0.473±0.047 Model group 0.304±0.069 0.311±0.052 Bone-Strengthening Group A 0.478±0.083 0.452±0.063 Bone-Strengthening Group B 0.425±0.061 0.407±0.055

[0105] The capsule of Example 1 of the present invention has a significant difference in its effect on improving the decrease in bone mineral density in osteoporotic rats compared with the capsule of Comparative Example 1 of the prior art.

[0106] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for preparing bone-strengthening capsules, characterized in that, In the preparation of bone-strengthening capsules, an NKA-9 type resin column was used to adsorb the raw material extract for preparing bone-strengthening capsules, and then an eluent was used for elution. The bone-strengthening capsules are made from the following ingredients: Epimedium 1251g, Rehmannia glutinosa 1656g, Drynaria fortunei 828g, Astragalus membranaceus 1251g, Salvia miltiorrhiza 828g, Auricularia auricula-judae 663g, Cucumber seeds 663g; The raw material extract for preparing bone-strengthening capsules was adsorbed using an NKA-9 resin column. The loading volume of the raw material extract was 10 BV, the flow rate of the raw material extract in the resin column was 5 mL / min, the eluent was an ethanol solution, the flow rate of the eluent in the resin column was 9 mL / min, the concentration of the eluent was 80%, and the amount of the eluent used was 6 BV.

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