Selaginella tamariscina extract, clathrate compound, Selaginella tamariscina formula granules and preparation method thereof

Abstract

The invention discloses a selaginella tamariscina extract, a selaginella tamariscina clathrate compound, selaginella tamariscina formula granules and a preparation method thereof, and relates to the field of medicines. The invention provides a preparation method of a selaginella tamariscina extract, which comprises the following steps: adding 12-16 times of water into selaginella tamariscina, soaking, decocting and distilling for the first time, and collecting first volatile oil and dregs; adding 8-10 times of water into the medicine residues, performing secondary decoction and distillation, and collecting second volatile oil and decoction liquid; combining the first volatile oil and the second volatile oil. The preparation process can effectively extract the active ingredients in the selaginella tamariscina, the active ingredients are applied to preparation of the selaginella tamariscina formula granules, the content and transfer rate of volatile oil in the selaginella tamariscina formula granules and the extraction rate of amentoflavone can be remarkably increased, so that the clinical effectiveness of the selaginella tamariscina formula granules is improved, and the selaginella tamariscina formula granules have the advantages of being high in stability, simple to operate, green, environmentally friendly and the like; and a way is provided for large-batch production of the selaginella tamariscina formula granules.

Description

Technical Field

[0001] The invention relates to the field of medicines, and in particular to a Selaginella extract, an inclusion compound and Selaginella formula particles and a preparation method thereof. Background Art

[0002] Selaginella is the dried whole herb of Selaginella pulvinata, also known as the resurrection grass and the pine of ten thousand years. It is a short herb with an upright main stem and clustered side branches in a rosette shape. The whole herb of Selaginella tastes light, slightly bitter, and cool in nature. Its functions and indications are to promote blood circulation and menstruation. It is used for amenorrhea, dysmenorrhea, lumps and masses, and injuries from falls. The volatile oil of Selaginella has anti-inflammatory, antioxidant, and anti-cancer activities.

[0003] Formula granules are a new type of Chinese herbal medicine slices, also known as decoction-free Chinese herbal medicine slices, refined decoction slices, scientific Chinese medicine, etc. They are made by extracting and concentrating single-flavor Chinese herbal medicine slices according to traditional standards. The advantage is that it is convenient for prescription preparation and carrying. The current process of making Selaginella granules is as follows: take Selaginella (cushion-shaped Selaginella) slices, add water to decoct, filter, concentrate the filtrate into a clear paste (dry soak), add appropriate amount of auxiliary materials, dry (or dry and crush), add appropriate amount of auxiliary materials, mix well, granulate, and make Selaginella granules.

[0004] The current Selaginella formula granules only contain water-decoction ingredients, and their therapeutic effect needs to be improved.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide a Selaginella extract, an inclusion compound and a Selaginella formula granule and a preparation method thereof.

[0007] The present invention is achieved in that:

[0008] In a first aspect, an embodiment of the present invention provides a method for preparing a Selaginella extract, comprising:

[0009] Adding 12 to 16 times the mass of water to the Selaginella, soaking, performing a first decoction and distillation, and collecting a first volatile oil and a medicinal residue;

[0010] Adding 8 to 10 times the weight of water to the medicinal residue, performing a second decoction and distillation, and collecting a second volatile oil and a decoction liquid;

[0011] The first volatile oil and the second volatile oil are mixed to obtain Selaginella volatile oil.

[0012] In a second aspect, an embodiment of the present invention provides a Selaginella extract, which comprises: at least one of the Selaginella volatile oil prepared by the preparation method described in the preceding embodiment, the decoction prepared by the preparation method described in the preceding embodiment, and the Selaginella extract in the decoction prepared by the preparation method described in the preceding embodiment.

[0013] In a third aspect, an embodiment of the present invention provides an inclusion compound containing the Selaginella extract described in the aforementioned embodiment.

[0014] In a fourth aspect, an embodiment of the present invention provides a method for preparing the inclusion compound as described in the aforementioned embodiment, comprising: preparing Selaginella volatile oil by the preparation method described in the aforementioned embodiment; mixing the Selaginella volatile oil with β-cyclodextrin and water to obtain the inclusion compound.

[0015] In a fifth aspect, an embodiment of the present invention provides a Selaginella formula granule, which includes: the Selaginella extract described in the aforementioned embodiment and / or the inclusion compound described in the aforementioned embodiment or the inclusion compound prepared by the preparation method described in the aforementioned embodiment.

[0016] In a sixth aspect, an embodiment of the present invention provides a method for preparing the Selaginella formula granules as described in the aforementioned embodiment, comprising: preparing Selaginella volatile oil using the preparation method described in the aforementioned embodiment, and preparing the Selaginella extract in the decoction using the preparation method described in the aforementioned embodiment; mixing the Selaginella volatile oil and the Selaginella extract in the decoction, crushing and sieving them, and then adding auxiliary materials to prepare the Selaginella formula granules.

[0017] The present invention has the following beneficial effects:

[0018] The preparation process provided by the present invention can more effectively extract the active ingredients in Selaginella. The preparation process can effectively extract the active ingredients in Selaginella. When applied to the preparation of Selaginella formula granules, the volatile oil content and transfer rate in the Selaginella formula granules and the extraction rate of sedum biflavonoids can be significantly improved, thereby improving the clinical effectiveness of the Selaginella formula granules. The invention has the advantages of high stability, simple operation, and green environmental protection, and provides a way for the mass production of Selaginella formula granules. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0020] Figure 1 A schematic diagram of a preparation process provided by an embodiment of the present invention;

[0021] Figure 2 This is the characteristic spectrum of Selaginella medicinal materials and formula granules. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0023] The existing standard quality standard of Selaginella formula granules is: 11000g of Selaginella slices are boiled in water, filtered, the filtrate is concentrated to a clear paste, an appropriate amount of auxiliary materials are added, dried, and then an appropriate amount of auxiliary materials are added, mixed, and granulated to 1000g. The current Selaginella formula granules only have water decoction ingredients. The content of selaginella biflavonoids is used to evaluate the content of Selaginella medicinal materials (the pharmacopoeia stipulates that the content of selaginella biflavonoids (C 30 H 18 O 10 ) shall not be less than 0.30%) and the content of Selaginella formula granules, the extraction rate of sedum biflavonoids is only 0.4% to 2.7% or lower, resulting in a waste of medicinal resources. The extraction rate calculation method: extraction rate % = extract content 0.14mg / (prescription 11g*medicinal material content 0.3%) / 1000*100%.

[0024] Based on the defects of the prior art, the embodiments of the present application provide a new method for preparing Selaginella extract and Selaginella formula granules. By setting specific process parameters, the extraction of active ingredients such as volatile oil and selaginella biflavonoids in Selaginella medicinal materials is effectively improved, thereby improving the utilization rate of Selaginella medicinal materials and the clinical therapeutic effect of Selaginella formula granules.

[0025] In one aspect, an embodiment of the present invention provides a method for preparing a Selaginella extract, comprising:

[0026] Adding 12 to 16 times the mass of water to the Selaginella, soaking, performing a first decoction and distillation, and collecting a first volatile oil and a medicinal residue;

[0027] Adding 8 to 10 times the weight of water to the medicinal residue, performing a second decoction and distillation, and collecting a second volatile oil and a decoction liquid;

[0028] The first volatile oil and the second volatile oil are mixed to obtain Selaginella volatile oil.

[0029] Through the preparation process under the above-mentioned limited conditions, it is possible to extract volatile oil from Selaginella medicinal materials more efficiently while extracting other effective ingredients in the medicinal materials. Compared with other processes, the preparation method provided in the embodiment of the present application is more stable and effective in increasing the content and transfer rate of Selaginella volatile oil and the transfer rate of sedum biflavonoids. Too low water volume is not conducive to the extraction of the index component sedum biflavonoids and other weakly polar components. Too much water volume will increase the content of the paste dilution unit, which is not conducive to large-scale industrial production and wastes manufacturing resources.

[0030] In some embodiments, the method of extracting volatile oils can be to connect a volatile oil extractor during decoction and distillation to collect and obtain volatile oils.

[0031] In some embodiments, the amount of water added to Selaginella is in the range of any one of 12, 13, 14, 15, 16 times or any two thereof relative to the mass of Selaginella.

[0032] In some embodiments, the amount of water added to the medicinal residue is in the range of 8, 9, 10 times or any two of the mass of the medicinal residue.

[0033] In some embodiments, the Selaginella comprises: Selaginella segments and / or Selaginella powder. The length of the Selaginella segments can be 5 to 10 mm, and can be any one of 5, 6, 7, 8, 9, and 10 mm, or any two thereof.

[0034] In some embodiments, the particle size of the Selaginella powder is ≤5 mm. Using Selaginella powder with a particle size of ≤5 mm is more conducive to the extraction of active ingredients.

[0035] In some embodiments, the soaking time is 20 to 60 minutes, and the soaking time can be any one of 20, 30, 40, 50, and 60 minutes, or a range between any two of them.

[0036] In some embodiments, the conditions of the first decoction and distillation include: 1 to 3 hours, 0.2 to 0.4 MPa, and 98 to 100°C.

[0037] In some embodiments, the conditions for the second decoction and distillation include: 1 to 3 hours, 0.2 to 0.4 MPa, and 98 to 100°C.

[0038] Decoction and distillation within the above-mentioned limited range is conducive to the efficient extraction of active ingredients of medicinal materials. If the time is shorter than the limited range, the active ingredients cannot be extracted, and if the time is longer than the limited range, resources will be wasted.

[0039] The time of the first decoction distillation or the second decoction distillation can be any one of 1, 2, and 3 hours or a range between any two of them, the pressure can be any one of 0.2, 0.3, and 0.4 MPa or a range between any two of them, and the temperature can be any one of 98, 99, and 100° C. or a range between any two of them.

[0040] In some embodiments, the preparation method further comprises salting out the mixture of the first volatile oil and the second volatile oil to obtain the Selaginella volatile oil.

[0041] In some embodiments, the salting out step comprises: adding sodium chloride to the mixture until saturated, filtering, and obtaining the Selaginella volatile oil. The filtering method can be to adjust the pressure in the suction bottle to -0.08 MPa, and filter with a 0.22 μm nylon 66 microporous filter membrane to obtain the Selaginella volatile oil.

[0042] In some embodiments, the preparation method further comprises: concentrating and / or drying the decoction to obtain the Selaginella extract in the decoction.

[0043] In some embodiments, the concentration method includes concentration under reduced pressure.

[0044] In some embodiments, the conditions for the reduced pressure concentration include: -0.072 to -0.03 MPa, 50 to 80°C, and the relative density at 60±5°C is 1.04 to 1.08 g / mL. The pressure can be any one of -0.072, -0.07, -0.06, -0.05, -0.04, and -0.03 MPa, or any two of them; the temperature can be any one of 50, 60, 70, and 80°C, or any two of them; and the relative density can be any one of 1.04, 1.05, 1.06, 1.07, and 1.08 g / mL, or any two of them.

[0045] In some embodiments, the drying method includes: spray drying or belt drying.

[0046] In some embodiments, the spray drying conditions include: the inlet air temperature is controlled at 160-180° C., the outlet air temperature is controlled at 70-100° C., and the feed frequency is 4-30 Hz.

[0047] In some embodiments, the conditions for belt drying include: tower vacuum is controlled at -100 to -99 kPa, zone one temperature is controlled at 80 to 100°C, zone two temperature is controlled at 60 to 80°C, zone three temperature is controlled at 20 to 40°C, crawler drive speed is controlled at 5 to 20 Hz, feed screw pump frequency is controlled at 1 to 15 Hz, and material distribution motor frequency is controlled at 7 to 15 Hz.

[0048] In some embodiments, before drying the decoction (including before concentration or after concentration and before drying), the preparation method further comprises filtering the decoction to obtain the filtrate.

[0049] Optionally, the decoction or its concentrated product is filtered using a 200-300 mesh filter cloth, and the mesh number of the filter cloth can be any one of 200, 220, 240, 260, 280, 300 mesh or a range between any two of them.

[0050] In some embodiments, after the decoction is concentrated and / or filtered and before drying, the preparation method further comprises: adding 0.1% to 2% maltodextrin by mass to the concentrated and / or filtered product and then drying to reduce spray drying loss. The amount of maltodextrin added can be any one of 0.1%, 0.5%, 1%, 1.5%, and 2% by mass of the product or a range between any two of them.

[0051] On the other hand, an embodiment of the present invention provides a Selaginella extract, which includes: at least one of: Selaginella volatile oil prepared by the preparation method described in any of the preceding embodiments, a decoction prepared by the preparation method described in any of the preceding embodiments, and a Selaginella extract in the decoction prepared by the preparation method described in any of the preceding embodiments.

[0052] On the other hand, an embodiment of the present invention provides an inclusion compound, which contains the Selaginella extract described in any of the above embodiments.

[0053] On the other hand, an embodiment of the present invention provides a method for preparing the inclusion compound as described in any of the above embodiments, comprising:

[0054] The volatile oil of Selaginella is prepared by the preparation method described in any of the above embodiments;

[0055] The Selaginella volatile oil is mixed with β-cyclodextrin and water to obtain an inclusion compound.

[0056] In some embodiments, the preparation method comprises: mixing β-cyclodextrin and water, and then mixing with Selaginella volatile oil.

[0057] In some embodiments, the added amount of β-cyclodextrin is 8 to 15 times the mass of the Selaginella volatile oil. Within this range, the saturation rate of the volatile oil is optimal. At the same time, pharmaceutical production requires minimizing the amount of excipients. Specifically, it can be any one of 8, 9, 10, 11, 12, 13, 14, and 15 times, or a range between any two of them.

[0058] In some embodiments, the amount of water added is 2 to 4 times the mass of the β-cyclodextrin. Too little water is not conducive to the dissolution of β-cyclodextrin and affects the inclusion rate of volatile oil. Too much water is not conducive to drying and wastes resources. The amount of water added can be any one of 2, 3, and 4 times or a range between any two of them.

[0059] In some embodiments, the method of mixing the Selaginella volatile oil with β-cyclodextrin and water comprises grinding.

[0060] In some embodiments, the grinding conditions include: 2000-3000 rpm, 15-35 min.

[0061] In some embodiments, the preparation method further comprises: drying the product obtained by mixing the Selaginella volatile oil with β-cyclodextrin and water to obtain a final inclusion complex.

[0062] In some embodiments, the drying method comprises vacuum drying.

[0063] In some embodiments, the drying conditions include: 55 to 75° C., -0.12 to -0.08 MPa. The temperature may be any one of 55, 60, 65, 70, 75° C. or any two thereof. The pressure may be any one of -0.12, -0.11, -0.10, -0.09, -0.08 MPa or any two thereof.

[0064] In some embodiments, after the Selaginella volatile oil is mixed with β-cyclodextrin and water, before drying, the preparation method further comprises refrigerating the mixed product. Refrigeration is conducive to the precipitation of β-cyclodextrin inclusion complex, and a relatively dry β-cyclodextrin inclusion complex is obtained by filtering, which is conducive to vacuum drying. It is also possible not to refrigerate, the amount of water is large, and the drying time is long.

[0065] In some embodiments, the refrigeration conditions are: time ≥ 12 hours, temperature 2-10° C. The time can be any one of 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, or a range between any two of them. The temperature can be any one of 2, 4, 6, 8, 10° C. or a range between any two of them.

[0066] On the other hand, an embodiment of the present invention provides a Selaginella formula granule, which includes: the Selaginella extract described in any of the preceding embodiments and / or the inclusion compound described in any of the preceding embodiments or the inclusion compound prepared by the preparation method described in any of the preceding embodiments.

[0067] In addition, an embodiment of the present invention also provides a method for preparing the Selaginella formula granules as described in any of the above embodiments, which comprises:

[0068] The Selaginella volatile oil is prepared by the preparation method described in any of the above embodiments, and the Selaginella extract in the decoction prepared by the preparation method described in any of the above embodiments;

[0069] The Selaginella volatile oil and the Selaginella extract in the decoction are mixed, crushed and sieved, and then auxiliary materials are added to prepare the Selaginella formula granules.

[0070] In some embodiments, the sieving includes: using a sieve with a mesh size of 80 to 100 meshes, which is beneficial to granulation uniformity and fluidity, and specifically can be any one of 80, 90, 100 meshes or a range between any two of them.

[0071] In some embodiments, after the Selaginella volatile oil and the Selaginella extract in the decoction are mixed, the preparation method further comprises drying the mixed product. The drying method and parameters can refer to the drying method and parameters described in any of the above embodiments, and will not be repeated here.

[0072] In some embodiments, the schematic diagram of the process of the Selaginella formula granules can refer to Figure 1 The inclusion compound and the Selaginella extract in the decoction can be dried first and then mixed (flow direction 1), or the inclusion compound and the Selaginella extract in the decoction can be mixed first and then dried (flow direction 2).

[0073] In some embodiments, the auxiliary material includes: maltodextrin. The amount of maltodextrin added is 0% to 1.18% of the mass of the mixture of the Selaginella volatile oil and the Selaginella extract in the decoction, and specifically can be any one of 0, 0.02%, 0.04%, 0.06%, 0.08%, 0.10%, 0.12%, 0.14%, 0.16%, 0.18%, or any two thereof. In some embodiments, the method for preparing the Selaginella formula granules includes dry granulation.

[0074] In some embodiments, the conditions for dry granulation include: roller speed of 1-4 rpm; extrusion pressure of 5-9 MPa; and feed speed of 41-60 rpm.

[0075] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0076] Example 1

[0077] A method for preparing Selaginella formula granules comprises the following steps.

[0078] 1. Weigh 11.1 kg of Selaginella truncatula powder (particle size of less than 5 mm) after crushing, add 12 times the weight of water to soak for 30 minutes, set 98 ° C, 0.3 MPa to decoct for 1.5 hours, and obtain the first volatile oil and medicinal residue; add 8 times the weight of water to the medicinal residue and set 98 ° C, 0.3 MPa to decoct for 1.0 hour to obtain the second volatile oil and decoction; the first volatile oil and the second volatile oil are combined as Selaginella truncatula volatile oil, with a total amount of 30.5 ml;

[0079] 2. The decoction was filtered through a 300-mesh filter bag and concentrated under reduced pressure at -0.06 to -0.04 MPa for 5 hours to obtain a concentrate with a specific gravity of 1.04 g / ml at 60°C. The concentrate was filtered through a 200-mesh filter and the filtrate was used as the Selaginella extract in the decoction.

[0080] 3. Add 10g of sodium chloride to the Selaginella volatile oil, stir and filter to obtain 28.4ml of volatile oil; add β-cyclodextrin and water to the Selaginella volatile oil, the amount of β-cyclodextrin added is 8 times the mass of the Selaginella volatile oil, the amount of water added is 2 times the mass of β-cyclodextrin, the colloid mill speed is 3200 rpm, the grinding time is 15 minutes, and the inclusion compound is obtained;

[0081] 4. The product of step 2 and the inclusion compound of step 3 were mixed, spray-dried, inlet air temperature: 180°C, outlet air temperature 80°C, feed frequency 12Hz, and 948.5g of dry powder was obtained;

[0082] 5. Dry granulation: Add 51.5 g of maltodextrin to the dry powder in step 4 to make Selaginella formula granules.

[0083] Example 2

[0084] A method for preparing Selaginella formula granules, comprising the following steps:

[0085] 1. Weigh 11.1 kg of Selaginella truncatula cut into short segments (length 5-10 mm), add 14 times the weight of water to soak for 30 minutes, set 98 ° C, 0.4 MPa to decoct for 2.0 hours to obtain the first volatile oil and medicinal residues; add 10 times the weight of water to the medicinal residues and set 98 ° C, 0.4 MPa to decoct for 2.0 hours to obtain the second volatile oil and decoction; the first volatile oil and the second volatile oil are combined as Selaginella truncatula volatile oil, with a total amount of 26.6 ml;

[0086] 2. The decoction liquid was filtered through a 200-mesh filter bag and concentrated under reduced pressure at -0.06 to -0.05 MPa for 6 hours to obtain a concentrate with a specific gravity of 1.04 g / ml at 60°C, filtered through a 200-mesh filter, and the filtrate was taken as the Selaginella extract in the decoction liquid. The Selaginella extract in the decoction liquid was added with 1% maltodextrin by mass of the Selaginella extract, stirred evenly, and spray-dried, with an inlet air temperature of 175°C, an outlet air temperature of 90°C, and a feed frequency of 10 Hz;

[0087] 3. Add 10g of sodium chloride to Selaginella volatile oil, stir and filter to obtain 23.4ml of volatile oil; add β-cyclodextrin and water to Selaginella volatile oil, the amount of β-cyclodextrin added is 10 times the mass of Selaginella volatile oil, the amount of water added is 4 times the mass of β-cyclodextrin, the colloid mill speed is 3000 rpm, the grinding time is 25 minutes, and the inclusion compound is obtained; refrigerate the inclusion compound for 12 hours, filter with filter paper, and dry the inclusion compound in a vacuum drying oven: set the temperature to 60°C and the vacuum degree to -0.08Mpa.

[0088] 4. Mix the products of step 2 and step 3, crush and pass through a 100 mesh sieve.

[0089] 5. Dry granulation: Add 130.5 g of maltodextrin to the product of step 4 (869.5 g) to prepare Selaginella formula granules.

[0090] Example 3

[0091] A method for preparing Selaginella formula granules comprises the following steps.

[0092] 1. Weigh 11.1 kg of Selaginella truncatula, which is a coarse powder (particle size is less than 5 mm) after crushing, add 14 times the weight of water to soak for 30 minutes, set 98 ° C, 0.2PMa to boil for 3 hours, and obtain the first volatile oil and the residue; add 12 times the weight of water to the residue and set 98 ° C, 0.2PMa to boil for 1.0 hour to obtain the second volatile oil and decoction; the first volatile oil and the second volatile oil are combined as the volatile oil of Selaginella truncatula, with a total amount of 29.7 ml;

[0093] 2. The decoction was filtered through a 300-mesh filter bag and concentrated under reduced pressure at -0.05 to -0.03 MPa for 5 hours to obtain a concentrate with a specific gravity of 1.08 g / ml at 60°C. The concentrate was filtered through a 200-mesh filter and the filtrate was used as the Selaginella extract in the decoction;

[0094] 3. Add 10g of sodium chloride to the Selaginella volatile oil, stir and filter to obtain 26.5ml of volatile oil; add β-cyclodextrin and water to the Selaginella volatile oil, the amount of β-cyclodextrin added is 12 times the mass of the Selaginella volatile oil, the amount of water added is 3 times the mass of β-cyclodextrin, the colloid mill speed is 2800 rpm, the grinding time is 30 minutes, and the inclusion compound is obtained;

[0095] 4. Mix the product of step 2 and the inclusion compound of step 3, vacuum dry, inlet air temperature: set temperature 65 ° C, vacuum degree minus 0.12Mpa, obtain dry powder, crush through a 100 mesh sieve, and mix well;

[0096] 5. Dry granulation: The dry powder (1008.2 g) of step 4 was made into Selaginella formula granules.

[0097] Comparative Example 1

[0098] A method for preparing Selaginella formula granules comprises the following steps.

[0099] 1. Weigh 11.1 kg of Selaginella slices (cut into sections), add water to cover the medicinal materials by 5 cm, soak for 30 minutes, set 98 ° C, 0.3 PMa to decoct for 1.0 hour, add water to the residue to 5 cm, set 98 ° C, 0.3 PMa to decoct for 30 minutes;

[0100] 2. The decoction liquid was filtered through a 300-mesh filter bag and concentrated under reduced pressure at -0.07 to -0.05 MPa for 5 hours to obtain a concentrated liquid with a specific gravity of 1.08 g / ml, which was then filtered through a 200-mesh filter;

[0101] 3. Add 3% maltodextrin and stir well, vacuum dry: set the temperature to 65°C and the vacuum degree to minus 0.12Mpa to obtain dry paste powder;

[0102] 4. Grind the dry paste through a 100-mesh sieve, mix well, and obtain dry powder (651 g).

[0103] 5. Dry granulation: add appropriate amount of maltodextrin to the dry powder in step 4 to prepare Selaginella formula granules.

[0104] Test Example 1 Determination of Volatile Oil

[0105] The volatile oil in the products of each stage of Examples 1 to 3 and Comparative Example 1 was determined, and the determination method (General Rules 2204 of the 2020 Edition of the Chinese Pharmacopoeia) is as follows.

[0106] Medicinal materials / decoction pieces: Weigh about 10g of the powder (passed through No. 2 to No. 3 sieves) (accurate to 0.01g), place in a flask, add 300-500ml (or appropriate amount) of water and a few glass beads, shake and mix, and connect the volatile oil analyzer and reflux condenser. Add water from the upper end of the condenser until it fills the scale part of the volatile oil analyzer and overflows into the flask. Place in an electric heating jacket and slowly heat to boiling, and keep it at a slight boil for about 5 hours until the amount of oil in the analyzer no longer increases, stop heating, leave it for a while, open the piston at the lower end of the analyzer, and slowly release the water until the upper end of the oil layer reaches 5mm above the scale 0 line. Leave it for more than 1 hour, then open the piston to let the oil layer drop until its upper end is just flush with the scale 0 line, read the amount of volatile oil, and calculate the content (%) of volatile oil in the test sample.

[0107] Selaginella formula granules: take appropriate amount of this product, grind it into powder, take 50g, and use the same method as the decoction pieces.

[0108] Test results

[0109] Table 1 Extraction rate of volatile oil components

[0110]

[0111]

[0112] Test Example 2 Determination of Azadirachta biflavonoids

[0113] The aconitum biflavonoids in the products of each stage of Examples 1 to 3 and Comparative Example 1 were determined.

[0114] Determination method

[0115] 1. Medicinal materials / decoction pieces

[0116] Chromatographic conditions and system suitability experiment: octadecylsilane bonded silica gel is used as filler; methanol is used as mobile phase A, 0.1% phosphoric acid solution is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; the detection wavelength is 330nm. The theoretical plate number calculated based on the peak of sedum biflavonoids should be no less than 3000.

[0117] Table 2 Elution program

[0118] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0～30 60 40 30～45 60→85 40→15

[0119] Preparation of reference solution: Take an appropriate amount of the flavonoids reference substance, accurately weigh it, and add methanol to make a solution containing 0.1 mg per 1 ml.

[0120] Preparation of test solution: Take about 0.2 g of the powder of the product (passed through No. 3 sieve), weigh it accurately, put it in a stoppered conical flask, add 50 ml of methanol accurately, weigh it, heat and reflux for 5 hours, cool, weigh it again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the solution.

[0121] Determination method: Accurately pipette 10μl of reference solution and 20μl of test solution respectively, inject into liquid chromatograph, and determine.

[0122] 2. Selaginella formula granules

[0123] The chromatographic conditions and system suitability experiments used octadecylsilane bonded silica as filler; acetonitrile-0.05% formic acid solution (acetonitrile and 0.05% formic acid solution were mixed in a volume ratio of 37:63) as the mobile phase for isocratic elution; the detection wavelength was 330nm. The theoretical plate number calculated based on the peak of the biflavonoids of Achyranthes bidentata should be no less than 3000.

[0124] Preparation of reference solution: Take an appropriate amount of the flavonoids reference substance, accurately weigh it, and add methanol to make a solution containing 20 μg per 1 ml.

[0125] Preparation of test solution: Take an appropriate amount of the product, grind it into powder, take about 0.2g, weigh it accurately, put it in a stoppered conical flask, accurately add 25ml of 70% ethanol, weigh it, heat and reflux for 30 minutes, cool, weigh it again, make up the lost weight with 70% ethanol, shake well, filter, and take the filtrate.

[0126] Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0127] Measurement results

[0128] Table 3 Extraction rate of flavonoids from edelweiss

[0129]

[0130] Test Example 3

[0131] Determine the fingerprint.

[0132] The fingerprint spectra of the Selaginella formula granules prepared in Examples 1 to 3 and Comparative Example 1 were determined according to high performance liquid chromatography (General Rules 0512 of the 2020 Edition of the Chinese Pharmacopoeia).

[0133] 1. Detection method:

[0134] Chromatographic conditions and system suitability experiment: octadecylsilane bonded silica gel is used as filler (column length is 150 mm; inner diameter is 2.1 mm; particle size is 1.6 um); acetonitrile is used as mobile phase A, 0.1% phosphoric acid solution is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; flow rate is 0.3 ml per minute; column temperature is 25°C; detection wavelength is 330 nm. The number of theoretical plates calculated based on the peak of sedum biflavonoids should be no less than 3000.

[0135] Table 4 Elution program

[0136] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0～2.5 5→10 95→90 2.5～7 10→15 90→85 7～12 15→18 85→82 12～14 18→20 82→80 14～24 20→47 80→53 24～26 47→60 53→40 26～27 60→90 40→10 27～29 90 10

[0137] Preparation of reference solution: Take an appropriate amount of the flavonoids reference substance, accurately weigh it, and add methanol to make a solution containing 0.1 mg per 1 ml.

[0138] Preparation of the test solution of medicinal materials / decoction pieces: Take about 3g of the powder of this product (passed through No. 3 sieve), add 25ml of methanol, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the solution.

[0139] Preparation of Selaginella formula granules for test sample: Take the product, grind it into powder, take 0.2g, add 25ml of methanol, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain.

[0140] Determination method: Accurately pipette 2 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0141] 2. Calculation method

[0142] The characteristic spectrum of Selaginella was used as the reference spectrum, and the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2.0)" was used to calibrate the 13 characteristic peaks of the test sample, and the similarity was calculated by Mark matching.

[0143] Test results see Figure 2 and Table 5.

[0144] Table 5 Similarity of fingerprints of each sample

[0145]

[0146] Taking the characteristic spectrum of Selaginella medicinal material as the reference spectrum, 13 characteristic peaks were matched using the Mark peak of the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2.0)". The similarities between the Selaginella formula granules 230801, 230802, and 230803 prepared in Examples 1 to 3 and the Selaginella medicinal material were 0.951, 0.957, and 0.968, respectively, while the similarity between the Selaginella formula granules 230804 prepared in Comparative Example 1 and the Selaginella medicinal material was 0.828. It can be seen that in addition to improving the transfer rate of the characteristic ingredient sedum biflavonoids in Selaginella medicinal materials and improving the utilization rate of medicinal materials, the present invention also takes into account the effective ingredients of the volatile oil of the medicinal materials, thereby improving the clinical effectiveness of the formula granules as Chinese herbal medicine pieces.

[0147] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a Selaginella extract, characterized in that: It includes: Adding 12 to 16 times the mass of water to the Selaginella, soaking, performing a first decoction and distillation, and collecting a first volatile oil and a medicinal residue; Adding 8 to 10 times the weight of water to the medicinal residue, performing a second decoction and distillation, and collecting a second volatile oil and a decoction liquid; The first volatile oil and the second volatile oil are mixed to obtain Selaginella volatile oil.

2. The preparation method according to claim 1, characterized in that: The Selaginella comprises: Selaginella segments and / or Selaginella powder; Optionally, the particle size of the Selaginella powder is ≤5 mm; Optionally, the soaking time is 20 to 60 minutes; Optionally, the conditions of the first decoction and distillation include: 1 to 3 hours, 0.2 to 0.4 MPa, 98 to 100° C.; Optionally, the conditions for the second decoction and distillation include: 1 to 3 hours, 0.2 to 0.4 MPa, 98 to 100° C.; Optionally, the preparation method further comprises salting out the mixture of the first volatile oil and the second volatile oil to obtain the Selaginella volatile oil; Optionally, the salting-out step comprises: adding sodium chloride to the mixture until saturated, and filtering to obtain the Selaginella volatile oil.

3. The preparation method according to claim 1 or 2, characterized in that: The preparation method further comprises: concentrating and / or drying the decoction to obtain the Selaginella extract in the decoction; Optionally, the concentration method includes concentration under reduced pressure; Optionally, the conditions for the reduced pressure concentration include: -0.072 to -0.03 MPa, concentrated to a relative density of 1.04 to 1.08 g / mL at 60±5°C; Optionally, the drying method includes: spray drying or belt drying; Optionally, before drying the decoction, the preparation method further comprises filtering the decoction; Optionally, the decoction is filtered using a 200-300 mesh filter cloth.

4. A Selaginella extract, characterized in that It includes: At least one of the Selaginella volatile oil prepared by the preparation method according to claim 1 or 2, the decoction prepared by the preparation method according to claim 1 or 2, and the Selaginella extract in the decoction prepared by the preparation method according to claim 3.

5. An inclusion compound, characterized in that: It contains the Selaginella extract according to claim 4.

6. The method for preparing the inclusion compound according to claim 5, characterized in that: It includes: The volatile oil of Selaginella is prepared by the preparation method according to claim 1 or 2; The Selaginella volatile oil is mixed with β-cyclodextrin and water to obtain an inclusion compound.

7. The preparation method according to claim 6, characterized in that: The preparation method comprises: mixing β-cyclodextrin and water, and then mixing with Selaginella volatile oil; Optionally, the added amount of the β-cyclodextrin is 8 to 15 times the mass of the Selaginella volatile oil; Optionally, the amount of water added is 2 to 4 times the mass of the β-cyclodextrin; Optionally, the method of mixing the Selaginella volatile oil with β-cyclodextrin and water comprises grinding; Optionally, the grinding conditions include: 2000-3000 rpm, 15-35 min; Optionally, the preparation method further comprises: drying the product obtained by mixing the Selaginella volatile oil with β-cyclodextrin and water to obtain a final inclusion compound; Optionally, the drying method includes vacuum drying; Optionally, the drying conditions include: 55 to 75°C, -0.08 to -0.12MPa; Optionally, after mixing the Selaginella volatile oil with β-cyclodextrin and water and before drying, the preparation method further comprises refrigerating the mixed product; Optionally, the refrigeration conditions are: time ≥ 12 h, temperature 2-10° C.

8. A Selaginella formula granule, characterized in that: It includes: The Selaginella extract according to claim 4 and / or the inclusion compound according to claim 5 or the inclusion compound prepared by the preparation method according to claim 6 or 7.

9. The method for preparing the Selaginella formula granules according to claim 8, characterized in that: It includes: The Selaginella volatile oil is prepared by the preparation method according to claim 1 or 2, and the Selaginella extract in the decoction is prepared by the preparation method according to claim 3; The Selaginella volatile oil and the Selaginella extract in the decoction are mixed, crushed and sieved, and then auxiliary materials are added to prepare the Selaginella formula granules.

10. The preparation method according to claim 9, characterized in that: The screening includes: using a sieve with a mesh size of 60 to 100 meshes; Optionally, the auxiliary material includes: maltodextrin.

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