CN115607515A

CN115607515A - Preparation process of spleen-tonifying miscarriage-preventing granules

Info

Publication number: CN115607515A
Application number: CN202211375487.2A
Authority: CN
Inventors: 章勤; 何嘉琳
Original assignee: Hangzhou Hospital of Traditional Chinese Medicine
Current assignee: Hangzhou Hospital of Traditional Chinese Medicine
Priority date: 2022-11-04
Filing date: 2022-11-04
Publication date: 2023-01-17
Anticipated expiration: 2042-11-04
Also published as: CN115607515B

Abstract

The invention belongs to the technical field of preparation of medicinal preparations, and particularly relates to a preparation process of spleen-tonifying miscarriage-preventing granules, which comprises the following steps: (1) extracting: decocting fifteen medicines with water twice, and filtering liquid medicine to obtain filtrate; (2) purification and concentration: concentrating the medicinal liquid to relative density of 1.05-1.10, standing, and collecting supernatant; (3) donkey-hide gelatin bead treatment addition: crushing donkey-hide gelatin beads, adding a proper amount of yellow wine, soaking overnight, heating and dissolving in water, filtering while hot, taking clear paste, adding the supernatant obtained in the step (2), and concentrating until the relative density is 1.20-1.25; (4) granulating: adding excipient and correctant, granulating, and drying. The invention strictly controls the preparation process of the spleen-tonifying miscarriage prevention granules and improves the quality of the spleen-tonifying miscarriage prevention granules.

Description

Preparation process of spleen-tonifying miscarriage-preventing granules

Technical Field

The invention belongs to the technical field of preparation of medicinal preparations, and particularly relates to a preparation process of spleen-tonifying miscarriage-preventing granules.

Background

The prescription for strengthening spleen and preventing miscarriage comes from many years of clinical experience and is improved from the He Shi miscarriage prevention drink. Is prepared from 16 Chinese medicaments of ramie root, baikal skullcap root, white paeony root with wine, medlar, chinese taxillus twig, largehead atractylodes rhizome, south dodder seed, yerbadetajo herb, pilose asiabell root, dried rehmannia root (charcoal), prepared rehmannia root (charcoal), perilla stem, liquoric root (fried), membranous milkvetch root (fried), himalayan teasel root and donkey-hide gelatin pearl, and is originally clinically used as a mixture packaged by multiple doses at present. In order to improve the defects of clinical use convenience and the like, the spleen-tonifying miscarriage prevention formula is developed for the second time, the use experience of a clinical mixture is fully kept on the selection of a dosage form and a process route, and the spleen-tonifying miscarriage prevention granules are developed on the basis of ensuring the drug effect and the safety, are easy to store and carry, more meet the requirements of current patients and are convenient for clinical use.

Disclosure of Invention

The preparation process of the spleen-tonifying miscarriage prevention granules is characterized in that the preparation process comprises the steps of extracting, concentrating, purifying, forming and the like.

Specifically, the technical scheme of the invention is as follows:

the invention provides a preparation process of spleen-tonifying miscarriage-prevention granules, which comprises the following steps:

(1) Extraction: ramie root, scutellaria baicalensis, white paeony root processed with wine, medlar, chinese taxillus twig, bighead atractylodes rhizome, semen cuscutae, eclipta, codonopsis pilosula, radix rehmanniae (charcoal), prepared rehmannia root (charcoal), perilla stem, liquorice (fried), astragalus (fried), teasel root and fifteen medicaments are decocted with water for two times, and the liquid medicine is filtered to obtain filtrate;

(2) And (3) purification and concentration: concentrating the medicinal liquid to relative density of 1.05-1.10, standing, and collecting supernatant;

(3) E, donkey-hide gelatin bead treatment: crushing donkey-hide gelatin beads, adding a proper amount of yellow wine, soaking overnight, heating and dissolving in water, filtering while hot, taking clear paste, adding the supernatant obtained in the step (2), and concentrating until the relative density is 1.20-1.25;

(4) Granulating: adding excipient and correctant, granulating, and drying.

Further, the step (1) is as follows: taking ramie root, scutellaria baicalensis, wine-processed white paeony root, medlar, loranthus parasiticus, bighead atractylodes rhizome, semen cuscutae, herba ecliptae, codonopsis pilosula, radix rehmanniae recen, prepared rehmannia root, perilla stem, liquorice (fried), astragalus mongholicus (fried) and teasel root into a round-bottom flask, adding 10 times of water into decoction pieces, decocting for 30-60 minutes, filtering, adding 6-8 times of lemon seed water into decoction dregs, decocting for 30-60 minutes, filtering, and combining filtrate.

Further, the step (3) is: pulverizing donkey-hide gelatin beads, sieving with a 40-mesh sieve, adding 5 times of yellow wine, soaking for 12 hours, heating over water to melt, filtering with a 100-mesh sieve while the mixture is hot, taking clear paste, adding the supernatant obtained in the step (2), and concentrating until the relative density is 1.20-1.25.

Further, the step (4) is: adopting fluidized bed spray granulation, the dosage of excipient is about 0.7-0.8 time of the solid content of the extract, and 0.25 percent of flavoring agent of the total amount of the granules is added.

Wherein the relative density of the concentration in the steps (2) and (3) is measured at 60-70 ℃.

Preferably, the preparation process of the spleen-tonifying miscarriage-prevention granule comprises the following steps:

(1) Extraction: putting ramie root, scutellaria baicalensis, white paeony root with wine, medlar, parasitic loranthus, bighead atractylodes rhizome, semen cuscutae, herba ecliptae, codonopsis pilosula, dried rehmannia root (charcoal), prepared rehmannia root (charcoal), perilla stem, liquorice (fried), astragalus mongholicus (fried) and teasel root into a round-bottom flask, adding 6-10 times of water into decoction pieces, decocting for 30-60 minutes, filtering, adding 6-8 times of lemon seed water into decoction dregs, decocting for 30-60 minutes, filtering, and combining filtrate;

(2) And (3) purification and concentration: concentrating the liquid medicine to the relative density of 1.05-1.10 at the temperature of 60-70 ℃, standing, and taking the supernatant;

(3) E, donkey-hide gelatin bead treatment: crushing donkey-hide gelatin beads, sieving with a 40-mesh sieve, adding 5 times of yellow wine, soaking for 12 hours, heating over water to melt the donkey-hide gelatin beads, filtering with a 100-mesh sieve while the donkey-hide gelatin beads are hot, taking clear paste, adding the supernatant obtained in the step (2), and concentrating until the relative density is 1.20-1.25 at 60-70 ℃;

(4) Granulating: adding dextrin as excipient in 0.7-0.8 times of the solid extract, adding sucralose in 0.25% of the total granule amount as corrective, spray granulating with fluidized bed, and drying.

Further, the lemon seed water in the step (1) is: soaking lemon seeds 1/5-1/10 times of the total weight of decoction pieces in water 20 times of the amount of the lemon seeds for 18-24 hr, and filtering.

Further, the lemon seed water has a pH =5.0-6.0.

Further, the medicinal materials in weight ratio are: 150-200 parts of ramie root, 90-130 parts of scutellaria baicalensis, 100-150 parts of white peony root with wine, 100-150 parts of wolfberry fruit, 150-200 parts of parasitic loranthus, 90-130 parts of bighead atractylodes rhizome, 100-150 parts of semen cuscutae, 100-150 parts of eclipta, 150-200 parts of codonopsis pilosula, 100-150 parts of radix rehmanniae recen, 100-150 parts of prepared rehmannia root, 50-100 parts of perilla stem, 30-75 parts of liquorice (fried), 160-210 parts of astragalus mongholicus (fried), 110-140 parts of teasel root and 85-140 parts of donkey-hide gelatin bead.

Preferably, the weight ratio of the medicinal materials is as follows: 187.5 parts of ramie root, 112.5 parts of scutellaria baicalensis, 125 parts of white peony root with wine, 125 parts of medlar, 187.5 parts of parasitic loranthus, 112.5 parts of bighead atractylodes rhizome, 125 parts of semen cuscutae, 125 parts of eclipta, 187.5 parts of codonopsis pilosula, 125 parts of radix rehmanniae (charred), 125 parts of prepared rehmannia root (charred), 75 parts of perilla stem, 50 parts of liquorice (fried), 187.5 parts of astragalus (fried), 125 parts of teasel root and 112.5 parts of donkey-hide gelatin pearl.

Compared with the prior art, the invention has the beneficial effects that:

(1) According to the requirements of the paste yield and the daily drinking amount of the decoction pieces, the preparation process of the spleen-tonifying miscarriage prevention granules is strictly controlled, the pretreatment method of the donkey-hide gelatin beads is optimized, the donkey-hide gelatin beads are added after being dissolved by yellow wine, so that the contents of L-hydroxyproline, glycine, alanine and L-proline and the contents of donkey-derived polypeptide A1 and donkey-derived polypeptide A2 are effectively improved, and the quality of the spleen-tonifying miscarriage prevention granules is improved;

(2) The invention optimizes the water extraction process, carries out water extraction on fifteen medicinal materials twice, and carries out decoction by adopting lemon seed water in the second water extraction, thereby improving the solid extraction rate and the total amount of astragaloside and further improving the quality of the spleen-tonifying miscarriage-prevention granules.

Drawings

FIG. 1: donkey-hide gelatin bead thin layer identification chromatogram

Wherein, 1, 2-negative samples with different elution amounts, 3, 4-samples with different elution amounts, the sample application amount is 20 mul

The sample application amount of 5-glycine, 6-L-hydroxyproline, 7-alanine and 8-proline is 1 mul

Developing agent: phenol-4% borax (4: 1) colour developer: 1% ninhydrin ethanol solution

FIG. 2: thin-layer discrimination chromatogram of white peony root with wine

Wherein, 1, 2-negative samples of different extraction methods, 3, 4-samples of different extraction methods

5-paeoniflorin, the sample application amount is 10 μ l

Developing agent: chloroform-ethyl acetate-methanol-formic acid (40: 5: 10: 0.2) color developer: 5% Vanillin sulfuric acid solution

FIG. 3: himalayan teasel root thin-layer identification chromatogram map

Wherein, the sample application amount of 1, 2-negative sample, 3, 4-sample is 2, 5, 2, 5 μ l 5-Dipsacus asperoides saponin VI, respectively, and is 10 μ l

Developing agent: lower layer solution developer of dichloromethane-methanol-water (14: 7: 2): 10% sulfuric acid ethanol solution

FIG. 4: thin layer chromatogram for identifying scutellaria baicalensis

Wherein, 1-negative sample is spotted in 8 μ l,2, 3 and 4-samples are spotted in 2, 5 and 8 μ l

5. 6-baicalin with the sample amount of 5 to 8 mul

Developing agent: ethyl acetate-acetone-dimethylformamide-glacial acetic acid-water (5: 2: 0.2: 0.3: 1)

Color developing agent: 2% ferric chloride ethanol solution.

FIGS. 5-8: partial chromatogram of content determination index of astragaloside used as spleen invigorating and miscarriage preventing granule

FIG. 5: astragaloside IV reference solution

FIG. 6: blank solvent

FIG. 7 is a schematic view of: negative solution of lack of astragalus root

FIG. 8: spleen-invigorating miscarriage-preventing granule test solution (acetonitrile-water 30: 70)

FIG. 9: spleen-invigorating miscarriage-preventing granule

Detailed Description

In order to make the purpose and technical solution of the present invention more clear, the present invention is further described with reference to the following examples, but the scope of the present invention is not limited to these examples, and the examples are only used for explaining the present invention. It will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true scope of the invention. 1. Research on preparation process of spleen-tonifying miscarriage-preventing granules

1. Process for treating donkey-hide gelatin beads in Chinese prescription

The donkey-hide gelatin beads are prepared by scalding donkey-hide gelatin with clam powder, and according to the traditional donkey-hide gelatin taking method, the main factors influencing the dissolution of the donkey-hide gelatin beads comprise the crushing granularity of the donkey-hide gelatin beads, a soaking solvent, the dosage and the like.

1.1 soak solvent selection

Weighing 3 parts of donkey-hide gelatin beads, each 100g of donkey-hide gelatin beads, crushing, sieving with a 40-mesh sieve, performing tests according to the arrangement in table 2, respectively soaking in purified water or yellow wine for 12 hours, and heating over water to dissolve the donkey-hide gelatin beads.

Observing the donkey-hide gelatin bead dissolving conditions in different soaking solvents and investigating the contents of the effective components of amino acids (L-hydroxyproline, glycine, alanine and L-proline) and donkey-derived polypeptides (donkey-derived polypeptide A1 and donkey-derived polypeptide A2), wherein the test results are shown in table 1.

TABLE 1 selection of solvents for Ejiao bead solubilization and investigation results of effective ingredient content of L-hydroxyproline and donkey-derived polypeptide

The results show that: the donkey-hide gelatin pearl uses yellow wine as a soaking solvent, has better dissolving effect, can improve the content of L-hydroxyproline, glycine, alanine and L-proline and the content of donkey-derived polypeptide A1 and donkey-derived polypeptide A2, and improves the quality of the spleen-tonifying miscarriage-prevention granules.

1.2 selection of crushing size

6 parts of donkey-hide gelatin beads are weighed, each part is 100g, tests are conducted according to the arrangement in the table 1, and after the donkey-hide gelatin beads with different grinding degrees are respectively soaked in yellow wine for a certain time, the yellow wine is separated from water and heated to be dissolved.

The melting condition of the donkey-hide gelatin beads with different crushing particle sizes is observed, and the test results are shown in table 2.

TABLE 2 examination of donkey-hide gelatin bead dissolution by crushing process

The results show that: the colla Corii Asini beads are pulverized, soaked in yellow wine, and heated in water to dissolve it effectively.

1.3 examination of solvent dosage

Weighing 4 parts of donkey-hide gelatin beads, each 100g of donkey-hide gelatin beads, crushing, sieving with a 40-mesh sieve, performing tests according to the arrangement in table 3, soaking in yellow wine for 12 hours, heating over water to dissolve, and filtering with a 100-mesh sieve while the yellow wine is hot.

The melting of the donkey-hide gelatin beads and the filtration of the clear paste are observed, and the test results are shown in table 3.

TABLE 3 investigation of donkey-hide gelatin bead dissolution by solvent dosage

The results show that: after the donkey-hide gelatin beads are crushed, yellow wine in an amount which is 5 times that of the crushed donkey-hide gelatin beads is selected as a soaking solvent, so that the dissolution effect is good, and the subsequent filtration process is facilitated.

And (3) combining the test results, and determining the pretreatment process of the donkey-hide gelatin beads in the formula as follows: pulverizing colla Corii Asini, sieving with 40 mesh sieve, soaking in 5 times of yellow wine for 12 hr, heating over water to dissolve, and hot filtering with 100 mesh sieve.

2. Research on water-adding extraction process

2.1 Effect of different decoction vessels on the extraction

The test method comprises the following steps: weighing 6 parts of decoction pieces with the batch number of 20211224, 15g of ramie root, 9g of scutellaria baicalensis, 10g of white peony root, 10g of medlar, 15g of parasitic loranthus, 9g of bighead atractylodes rhizome, 10g of dodder, 10g of eclipta, 15g of codonopsis pilosula, 10g of radix rehmanniae (charcoal), 10g of prepared rehmannia root (charcoal), 6g of perilla stem, 4g of fried liquorice, 15g of fried astragalus root and 10g of teasel root, placing the decoction pieces in different decocting containers, adding 10 times of water, heating and decocting for 60min, filtering, adding 10 times of water into dregs, decocting for 60min, filtering, combining the filtrates, concentrating to a proper amount, and extracting and concentrating respectively.

Respectively measuring the solid content in each test sample, calculating the average solid content extraction rate, and observing the influence of different decocting containers on the extraction of the medicinal materials by taking the solid content extraction rate as an index. The results are shown in Table 4.

TABLE 4 examination of extraction methods

The results show that: different decocting containers have great influence on the extraction rate of solid matters.

The solid extraction rate is directly influenced by the water evaporation amount due to the water evaporation during the extraction process when the decoction pot is adopted for decoction because of the tightness of the pot body.

When the round-bottom flask is adopted for hot reflux extraction, water is hardly lost, the extraction efficiency is relatively higher, and compared with a multifunctional extraction tank in actual production, the extraction principles of the round-bottom flask and the multifunctional extraction tank are consistent.

Therefore, when the water extraction process is studied, a round-bottomed flask with good sealing property is more suitable.

2.2 examination of Process parameters for Water extraction

According to the results of preliminary experiments, the present method is found to have a high paste yield. Referring to the requirements of management regulations of traditional Chinese medicine decoction rooms in medical institutions, and combining the current situation that the extraction times of the traditional Chinese medicine decoction room are 2 times in the original clinical application of the traditional Chinese medicine decoction room, the extraction times are fixed to be 2 times, and the water solvent, the extraction time and the water adding amount are mainly investigated in process investigation.

The preparation method comprises the steps of weighing 20211224 batch of decoction pieces, 30g of ramie root, 18g of scutellaria baicalensis, 20g of white peony root, 20g of medlar, 30g of loranthus parasiticus, 18g of bighead atractylodes rhizome, 20g of semen cuscutae, 20g of herba ecliptae, 30g of codonopsis pilosula, 20g of radix rehmanniae (charcoal), 20g of prepared rehmannia root (charcoal), 12g of perilla stem, 8g of liquorice (fried), 30g of astragalus (fried) and 20g of teasel root by adopting a single-factor test method, and performing a test according to a specified extraction method. Each test number is in parallel 2.

Taking the solid extraction rate and the total amount of astragaloside as indexes, and performing comprehensive scoring according to the weight of 1: 1, and preferably selecting the optimal extraction process. The test results are shown in Table 5.

Table 5 extraction of process parameter investigation results (n = 2)

The results show that: trial number 2 gave the highest composite score (797.76).

Comprehensively considering the aspects of time saving and energy saving of actual production, the water adding extraction process is determined as follows: adding 6-10 times of water into decoction pieces, decocting for 30-60 min, filtering, adding 6-8 times of lemon seed water (pH = 5.0-6.0) into residue, decocting for 30-60 min, filtering, and mixing filtrates.

2.3 Process verification test

The decoction pieces with the batch number of 20211224, 30g of ramie root, 18g of scutellaria baicalensis, 20g of white peony root with wine, 20g of medlar, 30g of parasitic loranthus, 18g of atractylodes macrocephala, 20g of dodder, 20g of eclipta, 30g of codonopsis pilosula, 20g of radix rehmanniae (charcoal), 20g of prepared rehmannia root (charcoal), 12g of perilla stem, 8g of liquorice (fried), 30g of astragalus (fried) and 20g of teasel root are respectively weighed, and three verification tests are carried out according to the preferable extraction process method of the test number 2.

The solid extraction rate and the total amount of astragaloside in each test number sample are respectively measured, and the test results are shown in table 6.

TABLE 6 results of the extraction Process verification test

The results show that: the test results of all the investigation indexes of the three batches of verification test samples are basically parallel, and the optimized extraction process method is proved to be feasible.

3. Research on purification and concentration process

3.1 purification Process Studies

In order to ensure that the substance basis of the spleen-tonifying miscarriage prevention granules is basically consistent with the application of the original clinical prescription, the purification process is researched by adopting a standing impurity removal mode.

The test method comprises the following steps: concentrating the extractive solutions to certain relative density, standing for a suitable time, collecting supernatant, and discarding the bottom precipitate to remove impurities. The main factors influencing the standing and impurity removal are the relative density of the concentrated liquid medicine and the standing time. Considering the actual production time schedule, the standing time of the extracted and concentrated liquid medicine is generally the standing overnight (about 18 hours), so the relative density of the liquid medicine is mainly considered.

Weighing decoction pieces with the batch number of 20211224, 45g of ramie root, 27g of scutellaria baicalensis, 30g of white peony root with wine, 30g of wolfberry fruit, 45g of parasitic loranthus, 27g of bighead atractylodes rhizome, 30g of dodder, 30g of eclipta, 45g of codonopsis pilosula, 30g of radix rehmanniae (charcoal), 30g of prepared rehmannia root (charcoal), 18g of perilla stem, 12g of fried liquorice and 45g of fried astragalus root and 30g of teasel root respectively, extracting according to a preferable extraction process, concentrating the liquid medicine to different relative density ranges, and standing for 18 hours.

The solid extraction rate and the total amount of astragaloside are respectively measured in each test number sample, the astragaloside transfer rate is calculated, and the test results are shown in table 7.

TABLE 7 examination of the relative Density of medicinal solutions

The results show that: after the liquid medicine is concentrated to a certain density and is kept stand overnight, the precipitate is removed, the solid content can be effectively reduced, and the impurity removal effect is good.

Comprehensively considering the solid extraction rate and the astragaloside transfer rate, the purification process is determined as follows: the solution was concentrated to a relative density of 1.05-1.10 (measured at 60 ℃ C.), and left to stand overnight (about 18 hours).

3.2. Research on concentration process

In the current practical production, the liquid medicine concentration generally adopts decompression concentration, and the concentration equipment is basically a single-effect or multi-effect concentrator.

As the main components of the medicinal herbs in the prescription do not belong to volatile or heat-sensitive components, the temperature can be controlled within the range of 60-70 ℃ during concentration for reduced pressure concentration.

In the pilot study, the differences of the index components in the concentrated solution after the concentration under reduced pressure at 60 ℃ and 70 ℃ were mainly examined, and the results are shown in Table 8.

TABLE 8 examination of the concentration temperature

The results show that: when the concentration temperature is 60-70 ℃, the content of the index component in the liquid medicine is not greatly influenced.

4. Research on preparation forming process

4.1 selection of granulation methods for granules

The conventional granulation methods of traditional Chinese medicines mainly comprise wet granulation, dry granulation, fluidized bed spray granulation and the like. The fluidized bed spray granulation is to complete the operations of mixing, granulating and drying materials in one step in fluidized bed equipment, and is a commonly used granulation molding technology at present.

The spray granulation of the fluidized bed can greatly reduce the amount of the auxiliary materials, the content of the extract in the final finished product particles can reach 50-70%, and the prepared particles have the advantages of uniform size, round appearance, good fluidity, high production efficiency and convenient automatic control. Meanwhile, the granulating process is finished in a closed granulator, so that the production process is not easy to be polluted, and the quality of finished products is ensured.

Therefore, the product selects fluidized bed spray granulation as a granulation method when the preparation is formed.

4.2 control of the relative Density of the drug solution during fluidized bed spray granulation

The process adopts a fluidized bed spray granulation method to directly obtain granules, so that the control of the relative density of the liquid medicine is very critical during granulation, and if the liquid medicine is too thin, the drying time of the granules is prolonged; if the liquid medicine is too thick, the spray head is easy to block.

After experimental exploration and production experience, the supernatant of the liquid medicine after standing overnight is taken, the pretreated donkey-hide gelatin bead clear paste is added, the mixture is continuously concentrated to clear paste with the relative density of 1.20 to 1.25 (measured at 60 ℃), and excipient is added, so that the granules meeting the quality requirement can be smoothly granulated.

4.3 selection of variety and amount of adjuvants and preparation of granules

In the research of the forming process, dextrin and sucralose are selected as auxiliary materials, fluidized bed granulation is carried out according to the auxiliary material proportion in the granule forming prescription in the table 9, and the granulation feasibility of each prescription is investigated.

TABLE 9 granule formulation

And (3) respectively comparing the indexes such as properties, formability, fluidity, hygroscopicity and the like of the prepared granular samples, thereby screening out the optimal auxiliary material proportion.

5. Feasibility of granulation forming process

Tests show that the total amount of the auxiliary materials of the No. 4 forming prescription is only 0.6 time of the solid content of the clear paste, so that the extract cannot be completely sprayed in the preparation process of the granules; the granules can be smoothly prepared by adding 1 time, 0.8 time and 0.7 time of total amount of fluid extract solid matter into adjuvants of No. 1, no. 2 and No. 3 molding prescription respectively.

5.1 examination of particle formation easiness and particle yield

The results of examining the granulation difficulty and the granule yield of the molding formulations No. 1, no. 2 and No. 3 are shown in Table 10.

TABLE 10 examination of particle formation ease and particle yield for different formulations

5.2 characterization of the Properties

The granules prepared by the molding recipes of No. 1, no. 2 and No. 3 were compared in appearance, color and taste, and the results are shown in Table 11.

TABLE 11 description of the properties

5.3 solubility assay

10g of the resulting granules were taken, heated to 200ml of water, stirred for 5 minutes, and immediately observed to be completely dissolved or slightly turbid. The results are shown in Table 12.

5.4 formability test

Three particle samples of 20g are taken and sequentially pass through a No. 1 sieve and a No. 5 sieve, particles which can pass through the No. 1 sieve but cannot pass through the No. 5 sieve are collected, the particles are weighed and calculated according to the following formula, and the investigation result is detailed in a table 12.

Molding ratio (%) = weight of granule after sieving/weight of granule before sieving × 100%

5.5 flowability investigation

The flowability of the pellets was compared by measuring the angle of repose using an FT-104B angle of repose measuring instrument.

Taking a proper amount of particles, naturally dropping the particles from a funnel with a certain height (H) onto a horizontal plate until no more particles flow out of the funnel, respectively reading the radius scales of the horizontal plate from two directions to obtain the diameter of the bottom surface of the formed cone, calculating the average value (L) of the diameter, and calculating the repose angle (theta) of the particles according to the following formula.

tanθ＝2H/L

The flowability of the granules obtained with the different molding formulations was investigated and is shown in Table 12.

5.6 investigation of hygroscopicity

The particles with the thickness of about 2mm are placed at the bottom of a flat weighing bottle which is dried to the constant weight, after the particles are precisely weighed, the flat weighing bottle is placed in a constant-temperature constant-humidity box under the conditions of the temperature of 25 ℃ plus or minus 1 ℃ and the humidity of 80 percent plus or minus 2 percent, a flat weighing bottle cover is opened, the flat weighing bottle cover is placed for 24 hours, the flat weighing bottle cover is taken out, the flat weighing bottle is rapidly and precisely weighed, the moisture wicking percentage is calculated according to the following formula, and the result is shown in a table 12.

Moisture absorption (%) = (weight of particles after moisture absorption-weight of particles before moisture absorption)/weight of particles before moisture absorption × 100%

TABLE 12 examination results of moldability, flowability, and hygroscopicity

As can be seen from the experimental results in tables 10, 11 and 12, the granules prepared by the molding prescriptions No. 1, no. 2 and No. 3 all meet the requirements of the "granule" item of the general rules of the preparations of the current edition of Chinese pharmacopoeia.

Comprehensively considering the research results of pilot plant test, the spleen-invigorating miscarriage-preventing granules are subjected to spray granulation by a fluidized bed, the dosage of excipient dextrin is about 0.7-0.8 time of the solid content of the extract, and sucralose accounting for 0.25 percent of the total amount of the granules is added as a flavoring agent.

6. Preparation process of spleen-tonifying miscarriage-preventing granules determined by pilot plant research

The process screened by the laboratory research is proved by the results of pilot scale research and sample trial production: the process is suitable for practical production.

6.1 prescription

6.2 preparation of

(1) Extraction: mixing radix Boehmeriae, scutellariae radix, radix Paeoniae alba preparata, fructus Lycii, herba Taxilli, atractylodis rhizoma, semen Cuscutae, ecliptae herba, radix Codonopsis, radix rehmanniae (charcoal), radix rehmanniae Preparata (charcoal), caulis Perillae, glycyrrhrizae radix (parched), radix astragali (parched), and radix Dipsaci to obtain round bottom flask, adding 10 times of water into decoction pieces, decocting for 30-60 min, filtering, adding 6-8 times of lemon seed water into residue, decocting for 30-60 min, filtering, and mixing filtrates;

(4) Granulating: adding dextrin as excipient in the amount of 0.7-0.8 times the solid content of the extract, adding sucralose accounting for 0.25% of the total amount of the granules as a flavoring agent, performing spray granulation by using a fluidized bed, and drying to prepare 1000g of the extract.

According to the usage amount (167 g decoction pieces/day) and the range of the extraction yield of the original clinical prescription, the preparation specification of the product is determined that 1g of granules is equivalent to 2.09g of decoction pieces.

2. Quality verification of spleen-tonifying miscarriage-preventing granules

Aiming at the research of a quality control method of the spleen-tonifying miscarriage-prevention granules, the prepared spleen-tonifying miscarriage-prevention granules are subjected to identification and content determination of medicinal materials such as donkey-hide gelatin beads, white paeony root, baical skullcap root, himalayan teasel root and the like.

1. Authentication

According to the prescription composition, the known main index components of the medicines are screened, and donkey-hide gelatin beads, white paeony root with wine, baical skullcap root and himalayan teasel root are preliminarily determined to be used as thin-layer identification indexes.

1.1 thin layer differential study of donkey-hide gelatin beads

Preparing a test solution, taking 8g of powder of the product, placing the powder into a 100ml anaerobic bottle, adding 30ml of 6mol/L hydrochloric acid, uniformly stirring, sealing, heating at 105 ℃ for 6 hours, taking out, cooling, adding 30ml of water, uniformly stirring, filtering, washing residues with 30ml of water, merging the residues into filtrate, adding a sodium hydroxide saturated solution into the filtrate to neutralize the filtrate until the pH is 5-6, concentrating the filtrate to 5-10 ml, adding the filtrate into a 732 strong acid styrene cation exchange resin column (the column length is 16cm, the inner diameter is 1.5cm, the pretreatment is H type), eluting with 100ml of water, discarding the eluent, adding 7ml of 3% sodium hydroxide for elution, collecting the eluent, evaporating to dryness, and adding 1ml of 30% ethanol into the residues for dissolving.

Preparation of control solution A glycine control was added with 30% ethanol to make a 1mg solution per 1 ml.

Sucking 20 μ l of sample solution and 1 μ l of reference solution under chromatographic conditions, respectively dropping on the same silica gel G thin layer plate, developing with phenol-4% borax (4: 1) as developing agent, taking out, air drying, spraying with 1% ninhydrin ethanol solution, and heating until the spots are clearly developed.

The method is characterized in that 8g of negative samples without donkey-hide gelatin beads are taken in a special test, the negative samples without donkey-hide gelatin beads are prepared into a negative test sample solution without donkey-hide gelatin beads by the method under the item of preparation of the test sample solution, no corresponding spots exist on the positions corresponding to the chromatogram of a glycine reference substance after development, which indicates that the negative samples without donkey-hide gelatin beads have no interference to the experiment (see figure 1), wherein the sample application amount of 1, 2-negative samples with different elution amounts, the sample application amount of 3, 4-samples with different elution amounts is 20 mul, 5-glycine, 6-L-hydroxyproline, 7-alanine, 8-proline, and the sample application amount is 1 mul, and the developing agent: phenol-4% borax (4: 1) colour developer: 1% ninhydrin ethanol solution.

1.2 thin layer identification of white peony root in wine

Preparing test solution by collecting powder 5g, adding ethanol 2ml and ethyl acetate 20ml, heating and refluxing for 30 min, cooling, filtering, evaporating filtrate to dryness, dissolving residue with ethanol 1ml, adding into neutral alumina column (3 g, inner diameter 1.5 cm), eluting with ethanol 30ml, collecting eluate, evaporating to dryness, and dissolving residue with ethanol 1 ml.

Preparation of control solution paeoniflorin control was added with methanol to make a solution containing 0.5mg per 1 ml.

Sucking 10 μ l of each of the test solution and the control solution under chromatographic conditions, respectively dropping on the same silica gel G thin layer plate, developing with chloroform-ethyl acetate-methanol-formic acid (40: 5: 10: 0.2) as developing agent, taking out, air drying, spraying with 5% vanillin sulfuric acid solution, and heating until the spots are clearly developed.

The specificity test takes 5g of the negative sample of the white peony root without wine, the negative sample of the white peony root without wine is prepared into the negative sample solution of the white peony root without wine by the method under the item of 'preparation of the sample solution', after the negative sample is unfolded, no corresponding spot exists on the position corresponding to the chromatogram of the paeoniflorin reference substance, which indicates that the negative sample of the white peony root without wine has no interference to the experiment (see figure 2), wherein: 1. 2-negative samples of different extraction methods, 3, 4-samples of different extraction methods 5-paeoniflorin, the sample application amount is 10 mul, developing agent: chloroform-ethyl acetate-methanol-formic acid (40: 5: 10: 0.2) color developer: 5% vanillin sulfuric acid solution.

1.3 thin layer identification of teasel root

Preparing test solution by collecting powder 5g, adding methanol 30ml, ultrasonic treating for 30 min, filtering, evaporating filtrate to dryness, dissolving residue with water 20ml, extracting with water saturated n-butanol 50ml, collecting n-butanol solution, washing with ammonia solution 30ml, washing with water 30ml, evaporating n-butanol solution to dryness, and dissolving residue with methanol 1 ml.

Preparation of reference solution radix Dipsaci saponin VI reference substance is prepared by adding methanol to obtain 1mg solution per 1 ml.

Sucking 2 and 5 μ l of each of the sample solution and the control solution under chromatography, respectively spotting on the same silica gel G thin layer plate, developing with dichloromethane-methanol-water (14: 7: 2) lower layer solution as developing agent, taking out, air drying, spraying 10% sulphuric acid ethanol solution, and heating until the spots are clearly developed.

The specificity test takes 5g of the negative sample of the lacking teasel root, the negative sample of the lacking teasel root is prepared into the negative sample solution of the lacking teasel root by the method under the item of the preparation of the sample solution, and no corresponding spot exists on the position corresponding to the chromatogram of the asperosaponin VI reference substance after the negative sample of the lacking teasel root is developed, which indicates that the negative sample of the lacking teasel root has no interference to the experiment (see figure 3). Wherein: 1. 2-negative sample, 3, 4-sample, sample application amount is 2, 5, 2, 5 mul, 5-dipsacoside VI, sample application amount is 10 mul, developing agent: lower layer solution developer of dichloromethane-methanol-water (14: 7: 2): 10% sulfuric acid ethanol solution.

1.4 thin layer identification study of Scutellaria baicalensis Georgi

The test solution is prepared by collecting powder 5g, adding ethyl acetate-methanol (3: 1) 30ml, heating under reflux for 30 min, cooling, filtering, evaporating filtrate, and dissolving residue with methanol 1 ml.

Preparation of control solution baicalin control was added with methanol to make 1mg solution per 1 ml.

Sucking 5 μ l of each of the sample solution and the control solution under chromatographic conditions, respectively dropping on the same silica gel G thin layer plate, developing with ethyl acetate-acetone-dimethylformamide-glacial acetic acid-water (5: 2: 0.2: 0.3: 1) as developing agent, taking out, air drying, and spraying with 2% ferric trichloride ethanol solution.

The specific test takes 5g of the negative sample lacking the scutellaria baicalensis, the negative sample lacking the scutellaria baicalensis is prepared into the negative sample solution lacking the scutellaria baicalensis by the method under the item of preparation of the sample solution, and no corresponding spot exists on the position corresponding to the baicalin reference substance chromatogram after the negative sample lacking the scutellaria baicalensis is unfolded, which indicates that the negative sample lacking the scutellaria baicalensis has no interference to the experiment (see figure 4). Wherein: 1-negative sample, 8 mul of sample application amount, 2, 3 and 4-samples, 2, 5, 8 mul of sample application amount, 5 and 6-baicalin sample application amount, 5 and 8 mul of sample application amount, developing agent: ethyl acetate-acetone-dimethylformamide-glacial acetic acid-water (5: 2: 0.2: 0.3: 1), colour developer: 2% ferric chloride ethanol solution.

2. Determination of content

According to the prescription composition, the content of astragaloside is used as a main index component for content determination. The method for measuring the content of astragaloside comprises the following steps:

in chromatographic condition and system adaptability test, octadecylsilane chemically bonded silica is used as a filler; acetonitrile-water (30: 70) was used as the mobile phase, and the detection was carried out by an evaporative light scattering detector. The number of theoretical plates is not less than 4000 calculated according to astragaloside IV peak.

Preparation of control solution A proper amount of astragaloside IV control is precisely weighed, and 80% methanol is added to make into solution containing 0.4mg per 1 ml.

Preparation of a test sample solution 20ml of a sample for process research is precisely measured, the sample is placed in a round-bottom flask, 2ml of ammonia water and 28ml of water are precisely added, the weight is weighed, the mixture is heated and refluxed for 1 hour, the mixture is cooled, the weight is weighed again, the weight loss is compensated by a solvent, shaking is uniformly carried out, 25ml of the mixture is precisely measured and evaporated to dryness, 15ml of water is added into residues for dissolving, the mixture is shaken and extracted for 3 times by using a water-saturated n-butyl alcohol solution, 25ml of the mixture is each time, the n-butyl alcohol solution is combined and evaporated to dryness, the residues are dissolved by 80% of methanol, the mixture is transferred into a 5ml measuring flask, 80% of methanol is added to the scales, shaking is uniformly carried out, filtering is carried out, and a subsequent filtrate is obtained.

The determination method comprises precisely sucking 2 μ l and 5 μ l of control solution and 20 μ l of test solution, injecting into liquid chromatograph, determining, and calculating by external standard two-point method logarithm method (FIG. 5-8).

3. Clinical research of spleen-tonifying miscarriage-preventing granules

In view of the unclear etiology and pathological mechanism of recurrent abortion, the treatment of the recurrent abortion still belongs to a clinical problem. The spleen-invigorating miscarriage-preventing granule is prepared by modifying the He Shi miscarriage-preventing drink into a granule dosage form. On the basis of guaranteeing the drug effect, the spleen-tonifying miscarriage-preventing granule is easy to store and carry, and more meets the requirements of the current patients. At present, the preparation is applied for decades, the coverage rate of the preparation in the patients with pregnancy is about 95%, and the beneficial groups have about 50 ten thousand times. The early clinical research shows that the fetus protection effective rate of the spleen-tonifying fetus protection granules for treating fetal irritability is 94.67 percent, which is far higher than that of progesterone injection for treating fetal irritability, and no adverse safety event occurs.

Claims

1. A preparation process of spleen-tonifying miscarriage-preventing granules is characterized by comprising the following steps:

(3) E, donkey-hide gelatin bead treatment: pulverizing colla corii asini beads, adding a proper amount of yellow wine, soaking overnight, heating to dissolve in water, filtering while hot, taking clear paste, adding the supernatant obtained in the step (2), and concentrating until the relative density is 1.20-1.25;

(4) Granulating: adding appropriate amount of excipient and correctant, granulating, and drying.

2. The process according to claim 1, wherein the step (1) is: taking ramie root, scutellaria baicalensis, wine-processed white paeony root, medlar, parasitic loranthus, bighead atractylodes rhizome, semen cuscutae, herba ecliptae, codonopsis pilosula, radix rehmanniae recen, prepared rehmannia root, perilla stem, liquorice (fried), astragalus mongholicus (fried) and teasel root into a round-bottom flask, adding 6-10 times of water into decoction pieces, decocting for 30-60 minutes, filtering, adding 6-8 times of lemon seed water into decoction dregs, decocting for 30-60 minutes, filtering, and combining filtrate.

3. The process according to claim 1, wherein the step (3) is: pulverizing donkey-hide gelatin beads, sieving with a 40-mesh sieve, adding 5 times of yellow wine, soaking for 12 hours, heating over water to melt, filtering with a 100-mesh sieve while the mixture is hot, taking clear paste, adding the supernatant obtained in the step (2), and concentrating until the relative density is 1.20-1.25.

4. The process according to claim 1, wherein the step (4) is: adopting fluidized bed spray granulation, the dosage of the excipient is about 0.7 to 0.8 time of the solid content of the extract, and a flavoring agent accounting for 0.25 percent of the total amount of the granules is added.

5. The process according to claim 1, wherein the relative density of the concentrate in steps (2) and (3) is measured at 60 ℃ to 70 ℃.

6. The process according to any one of claims 1 to 5, comprising the steps of:

(1) Extraction: mixing radix Boehmeriae, scutellariae radix, radix Paeoniae alba preparata, fructus Lycii, herba Taxilli, atractylodis rhizoma, semen Cuscutae, ecliptae herba, radix Codonopsis, radix rehmanniae (charcoal), radix rehmanniae Preparata (charcoal), caulis Perillae, glycyrrhrizae radix (parched), radix astragali (parched), and radix Dipsaci to obtain round bottom flask, adding 6-10 times of water into decoction pieces, decocting for 30-60 min, filtering, adding 6-8 times of lemon seed water into residue, decocting for 30-60 min, filtering, and mixing filtrates;

(4) And (3) granulating: adding dextrin as excipient in the amount of 0.7-0.8 times the solid content of the extract, adding sucralose accounting for 0.25% of the total amount of the granules as a flavoring agent, performing spray granulation by a fluidized bed, and drying to obtain the oral liquid.

7. The preparation process according to claim 2, wherein the lemon seed water in the step (1) is: soaking lemon seeds 1/5-1/10 times of the total weight of decoction pieces in water 20 times of the amount of the lemon seeds for 18-24 hr, and filtering.

8. The process of claim 2 or 7, wherein the lemon seed water has a pH =5.0-6.0.

9. The preparation process according to claim 1, wherein the weight ratio of the medicinal materials is as follows: 150-200 parts of ramie root, 90-130 parts of scutellaria baicalensis, 100-150 parts of white peony root with wine, 100-150 parts of wolfberry fruit, 150-200 parts of parasitic loranthus, 90-130 parts of bighead atractylodes rhizome, 100-150 parts of semen cuscutae, 100-150 parts of eclipta, 150-200 parts of codonopsis pilosula, 100-150 parts of radix rehmanniae recen, 100-150 parts of prepared rehmannia root, 50-100 parts of perilla stem, 30-75 parts of liquorice (fried), 160-210 parts of astragalus mongholicus (fried), 110-140 parts of teasel root and 85-140 parts of donkey-hide gelatin bead.

10. The preparation process according to claim 9, wherein the weight ratio of the medicinal materials is as follows: 187.5 parts of ramie root, 112.5 parts of scutellaria baicalensis, 125 parts of white peony root with wine, 125 parts of medlar, 187.5 parts of parasitic loranthus, 112.5 parts of bighead atractylodes rhizome, 125 parts of semen cuscutae, 125 parts of eclipta, 187.5 parts of codonopsis pilosula, 125 parts of radix rehmanniae (charred), 125 parts of prepared rehmannia root (charred), 75 parts of perilla stem, 50 parts of liquorice (fried), 187.5 parts of astragalus (fried), 125 parts of teasel root and 112.5 parts of donkey-hide gelatin pearl.