A preparation method of Platycodon decoction freeze-dried instant tablets

Abstract

The invention provides a preparation method of platycodon decoction freeze-dried instant tablets. The preparation method comprises the following steps: a, weighing raw materials with weight proportion: 4-6 parts of platycodon and 8-12 parts of liquorice; b, extracting: soaking in water, decocting, filtering, and obtaining filtrate; c, clarifying: adding a clarifier to the extract of step b, standing, filtering, and obtaining filtrate; d, freeze-drying: concentrating the filtrate of step c, and adding a flavoring agent, a freeze-dried skeleton agent, and a freeze-dried adhesive, wherein the amounts of the flavoring agent, the freeze-dried skeleton agent, and the freeze-dried adhesive are respectively: 1-5% of the flavoring agent, 2.0-8.0% of the freeze-dried skeleton agent, and 0.05-0.25% of the freeze-dried adhesive based on the filtrate after clarification in step c; freeze-drying temperature: the eutectic point is -11.03° C., and the eutectic point is -0.51° C.; and e, tableting. The freeze-dried instant dissolving tablets of the present invention are mainly taken by dissolving in water, and can be taken without water when necessary, thereby improving the convenience and compliance of use and promoting the development and innovation of new dosage forms of classic prescriptions.

Description

Technical Field

[0001] The invention relates to a preparation method of Platycodon decoction freeze-dried instant dissolving tablets, belonging to the field of traditional Chinese medicine preparations. Background Art

[0002] Throat discomfort is a common clinical symptom, mainly including dry throat, pain, itching and irritating cough, which seriously affects the patient's quality of life. In recent years, with the deterioration of air quality, serious air pollution, increased work pressure, more bad habits, different living habits, and diversified occupations, the incidence of throat diseases has increased year by year. The pathogenesis of this type of disease is complex and is related to upper respiratory tract inflammation, long-term alcoholism, dust in the air, and the stimulation of harmful gases. Modern medicine has no specific medicine for this type of disease, and the treatment method is mainly to prevent and relieve symptoms. In terms of lifestyle, patients are advised to quit smoking and drinking and eat a light diet. Clinically, most of the symptomatic treatments are mainly antibiotics and antiviral drugs, ultrasonic nebulization inhalation and other treatments.

[0003] Jie-Geng decoction (JGD) comes from the 311th article of Treatise on Febrile Diseases·Shaoyin by Zhang Zhongjing of the Eastern Han Dynasty: "Shaoyin disease, two or three days, sore throat, can be treated with Licorice decoction, if not cured, Jie-Geng decoction." It also states that it is mainly used to treat "lung abscess, cough and chest fullness, chills and rapid pulse, dry throat without thirst, occasional turbid saliva with fishy smell, and pus spitting out like rice porridge for a long time." It is widely seen in prescriptions of internal medicine, surgery, and pediatrics, especially various prescriptions for relieving exterior symptoms. A large number of clinical application reports show that Jie-Geng decoction is effective in treating throat diseases. For example, Yang Quansheng used Zibai Ganjue decoction to treat exogenous cough, Zhang Ling et al. used Jie-Geng decoction to treat chronic cough; Yu Jie used Jie-Geng Gancao decoction with added flavor to treat chronic throat darkening; Gao Shuying et al. treated acute bronchitis; Deng Sentian used Jie-Geng Gancao decoction to treat acute and chronic pharyngitis. In addition, Jie-Geng decoction also has the effects of clearing the lungs and relieving cough, removing phlegm and draining pus. Modern pharmacological research has found that Platycodon decoction has anti-inflammatory, apoptosis-inducing, airway mucin release-increasing, and anti-tumor effects. Its efficacy is related to ingredients such as platycodon saponins and glycyrrhizin in Platycodon decoction. Platycodon extract can effectively reduce inflammatory indicators and has a strong anti-inflammatory effect. It has been demonstrated that Platycodon can relieve pharyngeal mucosal inflammation, reduce the symptoms of pharyngitis, and prevent and treat chronic pharyngitis. Studies have also found that platycodon saponin D can inhibit the expression of inflammatory factors.

[0004] Platycodon decoction is still the most widely used and most effective decoction for the treatment of throat diseases, but traditional decoctions have disadvantages such as large oral dosage, inconvenience in carrying, easy to mold, and difficult to preserve. Summary of the invention

[0005] The present invention develops a new preparation of the classic prescription Jiegeng decoction to treat throat diseases, inherits and develops the therapeutic advantages of traditional Chinese medicine, improves existing problems, and develops a fast-dissolving formulation with the same efficacy as Jiegeng decoction.

[0006] The present invention provides a method for preparing Platycodon decoction freeze-dried instant tablets, which comprises the following steps:

[0007] a. Weigh the raw materials in the weight ratio:

[0008] 4-6 parts of Platycodon grandiflorum, 8-12 parts of Licorice root;

[0009] b. Extraction: Soak in water, decoct, filter and obtain the filtrate;

[0010] c. Clarification: add a clarifier to the extract in step b, let stand, filter, and obtain a filtrate;

[0011] d. Freeze-drying: After concentrating the filtrate in step c, add a flavoring agent, a freeze-dried skeleton agent, and a freeze-dried binder, wherein the amounts of the flavoring agent, the freeze-dried skeleton agent, and the freeze-dried binder are calculated based on the clarified filtrate in step c, respectively:

[0012] Flavor corrective 1-5%, freeze-dried skeleton 2.0-8.0%, freeze-dried adhesive 0.05-0.25%;

[0013] Freeze-drying temperature: eutectic point is -11.03℃, eutectic point is -0.51℃.

[0014] e. Tablet pressing.

[0015] Among them, the ratio of the raw materials described in step a is: 5 parts of Platycodon grandiflorum and 10 parts of Licorice; the concentration of the Platycodon grandiflorum decoction extract described in step b is 0.5g / mL.

[0016] Wherein, the clarifier described in step c is one or a mixture of two or more of chitosan, pectinase, PVPP and gelatin; the dosage of the clarifier is 0.5-5.0 g / L, and the standing time is 2-24 h.

[0017] Further preferably, the clarifier in step c is pectinase, the dosage is 3 g / L, and the standing time is 8 h.

[0018] Wherein, the flavoring agent in step d is HP-β-CD, α-CD, γ-CD, and the dosage is 1-5% based on the filtrate clarified in step c. Further preferably, the flavoring agent in step d is HP-β-CD, and the dosage is 3% based on the filtrate clarified in step c.

[0019] Among them, the freeze-dried skeleton agent described in step d is mannitol, maltodextrin, and sucrose, and the amount used is 2-8% based on the filtrate clarified in step c; the freeze-dried adhesive described in step d is mannitol, xanthan gum, gelatin, HPMC K4M, and pullulan, and the amount used is 0.10-0.20% based on the filtrate clarified in step c.

[0020] Further preferably, the freeze-dried skeleton agent in step d is mannitol, and the amount thereof is 5.06% based on the filtrate clarified in step c; the freeze-dried adhesive in step d is xanthan gum, and the amount thereof is 0.15% based on the filtrate clarified in step c.

[0021] Among them, the freeze-drying conditions described in step d are: pre-freezing temperature -50°C, pre-freezing time 3h, sublimation drying by gradient heating, starting from -50°C to 0°C, one gradient of 10°C, each gradient heating for 20min and maintaining for 40min, analytical drying temperature 25°C, analytical drying time 2h.

[0022] The invention also provides a preparation method of the Platycodon decoction freeze-dried instant tablets. The prepared Platycodon decoction freeze-dried instant tablets contain 0.919% to 0.924% of liquiritin, 3.116% to 3.136% of glycyrrhizic acid and 0.059% to 0.060% of platycodon saponin D.

[0023] The freeze-dried instant dissolving tablets of the present invention are mainly taken by dissolving in water, and can be taken without water when necessary, thereby improving the convenience and compliance of use and promoting the development and innovation of new dosage forms of classic prescriptions. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Visual analog rating chart;

[0025] Figure 2 Sublimation process diagram;

[0026] Figure 3 Full wavelength scan diagram;

[0027] Figure 4 Screening results of clarifier types (Note: B: Effect diagram of extract after clarification by clarifier, ①: pectinase, ②: chitosan, ③: PVPP, ④: gelatin);

[0028] Figure 5 Pectinase dosage results (Note: * indicates significant difference compared with Platycodon decoction extract, P<0.05);

[0029] Figure 6 Clarification time result diagram (Note: * indicates significant difference compared with Platycodon decoction extract, P<0.05);

[0030] Figure 7 Taste standard curve diagram (Note: A: astringency standard curve, B: bitterness standard curve, C: sweetness standard curve);

[0031] Figure 8 Radar chart (A) and box plot (B) of taste analysis;

[0032] Fig. 9Three-dimensional effect diagram (A) and contour diagram (B) of each factor on the comprehensive score;

[0033] Fig.10 DSC curve graph;

[0034] Fig.11 Cooling curve diagram;

[0035] Fig.12 Heating curve diagram;

[0036] Fig.13 Freeze-drying curve diagram. DETAILED DESCRIPTION

[0037] Example 1 Screening test of the preparation process of freeze-dried instant tablets of Jingfang Jiegeng Decoction

[0038] 1 Materials and Instruments

[0039] 1.1 Experimental Materials

[0040] The experimental materials used in this experiment are shown in Table 1:

[0041] Table 1 Main reagents and manufacturers

[0042]

[0043] Table 1

[0044]

[0045] 1.2 Instruments and Equipment

[0046] The main instruments and equipment used in this experiment are shown in Table 2:

[0047] Table 2 Main instrument models and manufacturers

[0048]

[0049]

[0050] 2 Experimental methods

[0051] 2.1 Preparation method of extract of traditional Chinese medicine Jiegeng decoction

[0052] Take 5g of Platycodon grandiflorum slices and 10g of Licorice slices, add 14.28 times water and soak for 1h, boil over high heat and then simmer over low heat for 120min. After decocting, filter with 200-mesh filter cloth and squeeze the medicinal materials to collect the decoction, combine the filtrate, place it in a rotary evaporator with a vacuum degree of 0.08Mpa and a temperature of 60℃ and reduce the pressure to concentrate to a liquid concentration of 0.5g / mL.

[0053] 2.2 Study on the clarification process of extract from traditional Chinese medicine Jiegeng decoction

[0054] 2.2.1 Determination of measurement wavelength

[0055] The full wavelength scanning of Platycodon grandiflorum extract solutions diluted 10 times, 20 times, 30 times and 40 times was performed by UV-visible spectrophotometer in the range of 200nm-1000nm, and distilled water was used as blank control.

[0056] 2.2.2 Screening of clarifier types

[0057] (1) Preparation of clarifier

[0058] 1% chitosan solution: accurately weigh 1.0 g chitosan and dissolve it in 100 mL of 0.2% citric acid solution, heat and boil until completely dissolved, cool and dilute to a 100 mL volumetric flask for later use.

[0059] 1% pectinase solution: Accurately weigh 1.0g of pectinase, add 50mL of 40-50℃ distilled water to dissolve, and prepare a 1% solution for use.

[0060] 1% PVPP solution: Dissolve 1.0 g PVPP in 50 mL distilled water, then dilute to 100 mL in a volumetric flask for later use.

[0061] 1% gelatin solution: Accurately weigh 1.0 g of gelatin in 50 mL of distilled water, soak for 24 hours, dissolve in a 40°C water bath, cool and dilute to a 100 mL volumetric flask for later use.

[0062] (2) Measurement method

[0063] Take 4 groups of stoppered test tubes, 5 tubes in each group, and add appropriate amount of Jiegeng Decoction extract to each test tube. Add different clarifiers to each group of test tubes. The specific concentrations of clarifiers are (g / L): chitosan: 0, 0.5, 1.0, 1.5, 2.0; pectinase: 0, 0.5, 1.0, 1.5, 2.0; PVPP: 0, 0.5, 1.0, 1.5, 2.0; gelatin: 0, 0.5, 1.0, 1.5, 2.0. After fully stirring, plug the test tubes, let them stand at room temperature for 10 hours, take the supernatant and measure its transmittance at 660nm, and use distilled water as blank control.

[0064] 2.2.3 Pectinase dosage screening

[0065] Take 11 test tubes with stoppers, add appropriate amount of Jiegeng Tang extract to each test tube, add different concentrations of pectinase to each test tube, the specific concentrations are (g / L): 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0. Stir well and stopper the test tubes, let stand at room temperature for 10 hours to clarify, take the supernatant and measure its transmittance at 660nm, and use distilled water as a blank control.

[0066] 2.2.4 Determination of clarification time

[0067] Take 9 test tubes with stoppers and add an equal amount of Jiegeng Tang extract containing 3.0g / L pectinase to each test tube. Stir thoroughly and plug the test tubes. Let them stand at room temperature for 0, 2, 4, 6, 8, 10, 12, 18, and 24 hours. Take the supernatant and measure its transmittance at 660nm, using distilled water as a blank control.

[0068] 2.3 Study on the flavor correction process of extract of traditional Chinese medicine Jiegeng decoction

[0069] 2.3.1 Screening and training of volunteers

[0070] The screening range of volunteers is: aged 18 to 28 years old, healthy people with no history of genetic diseases or recent medical history, no drinking or smoking 1 day before the experiment, and fasting but not drinking water for 2 hours before the experiment.

[0071] The solutions prepared with tannic acid, quinine sulfate and sucrose were used as the standard solutions for astringency, bitterness and sweetness. Tannic acid solutions at 1.0, 3.0 and 8.0 mg / mL, quinine sulfate solutions at 125, 375 and 1000 ng / mL and sucrose solutions at 100, 300 and 800 mg / mL were prepared respectively. The corresponding astringency, bitterness and sweetness scores were determined to be 2, 5 and 10 points through preliminary tests (see Figure 1 ). The volunteers tasted the sample solutions one by one and evaluated them using the visual analogue scoring (VAS). The volunteers were informed and asked to remember the taste scores corresponding to the three concentration gradients of different solutions. Then a single-blind test was conducted, and a sample solution was randomly selected from the above different solutions. If the volunteer tasted the solution correctly, the training was passed and a formal sensory test was conducted. Finally, 16 healthy volunteers were selected as subjects (6 male volunteers and 10 female volunteers, aged 18 to 28 years old).

[0072] 2.3.2 Volunteer Tasting Evaluation Method

[0073] The volunteers took 20 mL of sample solution and put it in their mouths, gargled in the mouth, so that the tongue and mouth can fully feel the taste of the medicine, and performed VAS scoring based on the changes in facial micro-expressions. After the sample enters the mouth, the taste can be fully felt in about 15 seconds, so the score is selected at the 15th second of the entrance, and then the liquid is spit out, and the mouth is rinsed with pure water 5 times until there is no residual taste in the mouth. The next sample is evaluated after 15 minutes. In this process, multiple tastes such as bitterness, astringency, and sweetness need to be scored, with scores ranging from 0 to 10 points.

[0074] The data were tested and eliminated for outliers using the Grubb test at a test level of 0.05. After eliminating outliers, the data were calculated as follows.

[0075] The calculation formula for each taste score (I) is as follows.

[0076] I=(I1+I2+I3+...+In) / n Formula 1-1

[0077] Where, I1, I2, …, In represent the unpleasant taste value given by each volunteer, and n represents the number of volunteers participating in the sensory evaluation. 2.3.3 Establishment of taste standard curve

[0078] (1) Establishment of astringency standard curve

[0079] Weigh an appropriate amount of tannic acid to prepare 0.125, 0.5, 1.0, 2.0, 4.0, 6.0, and 8.0 mg / mL tannic acid solutions as standard bitter solutions, and set the corresponding astringency scores to 0, 1, 2, 4, 6, 8, and 10 points. Take the above standard solutions for oral testing according to the method under 2.3.2, record the corresponding taste scores, draw a standard curve with the tannic acid solution concentration as the horizontal axis (X) and the corresponding taste score as the vertical axis (Y), and calculate the fitting equation.

[0080] (2) Establishment of bitterness standard curve

[0081] Weigh an appropriate amount of quinine sulfate to prepare 15.625, 62.5, 125, 250, 500, 750, and 1000 ng / mL quinine sulfate solutions as standard bitter solutions, and set the corresponding astringency scores to 0, 1, 2, 4, 6, 8, and 10 points. Take the above standard solutions for oral testing according to the method under 2.3.2, record the corresponding taste scores, draw a standard curve with the quinine sulfate solution concentration as the horizontal axis (X) and the corresponding taste score as the vertical axis (Y), and calculate the fitting equation.

[0082] (3) Establishment of sweetness standard curve

[0083] Weigh an appropriate amount of sucrose to prepare 12.5, 50, 100, 200, 400, 600, and 800 mg / mL sucrose solutions as standard sweetness solutions, and set the corresponding sweetness scores to 0, 1, 2, 4, 6, 8, and 10 points. Take the above standard solutions for oral testing according to the method under 2.3.2, record the corresponding taste scores, draw a standard curve with the concentration of tannic acid solution as the horizontal axis (X) and the corresponding taste score as the vertical axis (Y), and calculate the fitting equation.

[0084] (4) Volunteers selected the most suitable taste range

[0085] Take each of the above taste standard solutions and conduct oral test according to the method in 2.3.2, record the corresponding taste score, and record the most suitable taste score range of the volunteers, and use this as the target to screen the flavor corrective and dosage.

[0086] 2.3.4 Taste analysis of Platycodon grandiflorum decoction extract

[0087] Take 20 mL of Platycodon grandiflorum extract and put it in your mouth. Perform an oral test according to the method in 2.3.2 and record the corresponding taste score.

[0088] 2.3.5 Screening of flavoring agents

[0089] At present, the methods of correcting the flavor of traditional Chinese medicine are mostly adding single sweeteners, compound sweeteners and inclusion methods. The unique taste of licorice in Platycodon decoction has the effect of neutralizing bitterness to a certain extent. Therefore, the main purpose of correcting the flavor in this experiment is to correct the astringency. Simply adding sweeteners is not applicable, so the cyclodextrin inclusion method is chosen to correct the astringency.

[0090] According to Table 3, different types of cyclodextrin were added to the Platycodon decoction extract, and oral test was performed according to the method in 2.3.2, and the corresponding taste scores were recorded.

[0091] Table 3 Cyclodextrin preparation concentration

[0092]

[0093] 2.3.6 Study on the dosage of hydroxypropyl beta-cyclodextrin

[0094] Take 20 mL of Platycodon grandiflorum decoction extract, add 1.0%, 2.0%, 3.0%, 4.0%, 5.0% HP-β-CD respectively, stir in a 60℃ hot water bath for 30 min to dissolve, conduct oral test according to the method in 2.3.2, and record the corresponding taste scores.

[0095] 2.4 Prescription process screening of freeze-dried instant tablets of Jingfang Jiegeng Decoction

[0096] 2.4.1 Quantitative Rating Table

[0097] (1) Taste Rating Sheet

[0098] According to Table 4, the taste and mouthfeel were scored according to the corresponding evaluation methods and scoring standards.

[0099] Table 4 Tasting score table

[0100]

[0101] (2) Appearance Rating Table

[0102] According to Table 5, the appearance is scored for fullness, smoothness, color uniformity, degree of melting and formability according to the corresponding evaluation methods and scoring standards.

[0103] Table 5 Appearance rating table

[0104]

[0105] (3) Disintegration time scoring table

[0106] According to Table 6, the disintegration time was scored in vitro (disintegration apparatus method) and in vivo (volunteer oral tasting method) according to the corresponding evaluation methods and scoring criteria.

[0107] Table 6 Disintegration time scoring table

[0108]

[0109] 2.4.2 Screening of auxiliary materials

[0110] (1) Screening of freeze-dried matrix agents

[0111] The freeze-dried skeleton agent, also known as a filler, is used to support the formation of a loose and porous skeleton structure of the freeze-dried tablet and to maintain a certain hardness of the tablet. It can also be used as a freeze-dried protective agent to maintain the stability of the active ingredients during the freeze-drying process. According to Table 7, freeze-dried skeleton agents of different types and concentrations were added to the Platycodon decoction extract to examine its appearance and disintegration time.

[0112] Table 7 Lyophilized skeleton preparation concentration

[0113]

[0114] (2) Screening of freeze-dried adhesives

[0115] The freeze-dried tablets prepared by using the skeleton alone are prone to cracking or fragmentation, so they are often used in combination with a binder. According to Table 8, freeze-dried binders of different types and concentrations were added to the Platycodon decoction extract to examine its appearance and disintegration time.

[0116] Table 8 Freeze-dried adhesive preparation concentration

[0117]

[0118] 2.4.3 Comprehensive Rating

[0119] (1) Determine subjective weights using the AHP method

[0120] The three evaluation indicators of taste, appearance and disintegration time in the prescription are quantified as weight indicators, and the indicators are compared pairwise according to their importance to construct a priority judgment matrix, so as to calculate the weight coefficient of each indicator, which is the subjective weight WAHP.

[0121] (2) CRITIC method to determine objective weights

[0122] In order to eliminate the influence of different dimensions on the evaluation results, it is necessary to first use the normalization method to de-dimensionalize the experimental data of each indicator, and then calculate the comparison strength (S j ), index conflict (R j ), information volume (C j ), and finally calculate the objective weight of each indicator, namely W CRITIC .

[0123] (3) AHP-CRITIC subjective and objective weighting method to determine the comprehensive weight

[0124] The above subjective weight W AHP and objective weight W CRITIC The coefficient is calculated according to formula 1-2 to obtain the comprehensive weight, that is, W AHP-CRITIC .

[0125]

[0126] (4) Calculation of comprehensive scores of multiple indicators in the experimental plan

[0127] After normalizing the indicator data in the experimental plan according to formula 1-3, calculate according to formula 1-4 and 1-5 to obtain the comprehensive score.

[0128]

[0129] C ij =x ij *w ( A HF-CRITIC)j Formula 1-4

[0130]

[0131] Among them, x ij represents the value of the jth indicator of the i-th sample, C ij represents the weighted score of the jth indicator of the i-th sample, C i Represents the comprehensive score of the experimental scheme.

[0132] 2.4.4 Investigation of the amount of auxiliary materials

[0133] (2) Mannitol dosage study

[0134] Take 2.5g of Platycodon decoction extract, add 3.0% HP-β-CD and stir in a 60℃ hot water bath for 30min to dissolve; add 0.15% xanthan gum and let it swell overnight; add 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0% mannitol respectively and stir to mix. Determine the taste score, appearance score and disintegration time score according to the value score table under 2.4.1 respectively, calculate the comprehensive score by the method under 2.4.3, and perform one-way analysis of variance.

[0135] (2) Investigation of xanthan gum dosage

[0136] Take 2.5g of Platycodon decoction extract, add 3.0% HP-β-CD and stir in a 60℃ hot water bath for 30min to dissolve; add 0.05%, 0.10%, 0.15%, 0.20%, 0.25% xanthan gum respectively, let it stand to swell overnight; then add 5.0% mannitol and stir to mix. Determine the taste score, appearance score and disintegration time score according to the quantitative score table under 2.4.1 respectively, calculate the comprehensive score using the method under 2.4.3, and perform one-way analysis of variance.

[0137] 2.4.5 Optimization and screening of preparation prescriptions

[0138] (1) Central point design-effect surface method factor level design

[0139] According to the above single-factor experimental results, the mass concentrations of mannitol (X1) and xanthan gum (X2) in the prescription were taken as the investigation factors, and the optimization ranges of the two were 1.0%-8.0% and 0.05%-0.25%, respectively. The comprehensive score of taste score, appearance score and disintegration time score was taken as the response variable. A two-factor five-level star-point design experiment was adopted. The factors and levels are shown in Table 9.

[0140] Table 9 Factors and levels

[0141]

[0142] (2) Calculation of comprehensive score

[0143] The taste score, appearance score and disintegration time score were determined according to the quantitative score table in 2.4.1. The comprehensive score was calculated using the method in 2.4.3 and a one-way analysis of variance was performed.

[0144] (3) Central-site design-response surface methodology fitting

[0145] The relevant data were imported into Design-Expert 13.0 software, and the fitting equation was obtained by analysis. The correlation coefficient (r 2) and confidence (P) are used as model judgment criteria.

[0146] (4) Model optimization and verification

[0147] A three-dimensional effect surface diagram and a two-dimensional contour map were drawn based on the fitting equation model. Taking the maximum comprehensive score as the goal, three representative prescriptions were selected and their taste scores, appearance scores and disintegration time scores were measured according to the quantitative score table in 2.4.1. The comprehensive score was calculated using the method in 2.4.3, and the deviation was used to evaluate the predictive ability of the model.

[0148] 2.5 Study on the freeze-drying process of the freeze-dried instant tablets of Jingfang Jiegeng Decoction

[0149] 2.5.1 Determination of critical temperature

[0150] The eutectic point of a material refers to the temperature at which the water in the mixed solution is completely frozen into ice crystals, and the eutectic point refers to the temperature at which the frozen material begins to melt when heated. Differential scanning calorimetry (DSC) was used to measure the eutectic point and eutectic point of the raw material solution of the Platycodon decoction preparation before freeze-drying. Accurately weigh 3.1500 mg of the raw material solution of the Platycodon decoction preparation before freeze-drying and put it into a sealed aluminum box, cool it from 30°C to -50°C at a rate of 10°C / min and keep it for 5 minutes, and then scan it from -50°C to 30°C at a rate of 10°C / min.

[0151] 2.5.2 Determination of sublimation endpoint

[0152] The sublimation endpoint is determined by observing the waterline method, such as Figure 2 As shown, during the freeze-drying process, water sublimates from top to bottom. Therefore, during the sublimation drying process, as time goes by, the boundary line between the drying layer and the freezing layer (sublimation line) will also move from top to bottom; when the sublimation line completely reaches the bottom of the bottle and disappears, and then maintains for a period of time, the time at this time can be regarded as the end point of sublimation.

[0153] The method for determining the end point of sublimation drying is shown in Table 10. The cold trap temperature and silicone oil inlet and outlet temperatures are suitable for large and fully loaded samples, and the silicone oil inlet and outlet temperature method requires the installation of temperature probes at the silicone oil inlet and outlet. Other methods for determining the end point of sublimation drying that also require the installation of temperature sensors, pressure testers, capacitance vacuum gauges and other sensors are not applicable to laboratory small freeze dryers. Therefore, the only methods for determining the end point of sublimation drying used in laboratory small freeze dryers are: observing the waterline method, plate layer temperature and online sampling method. Although the online sampling method is applicable to the entire freeze-drying process, the sampling process requires the freeze dryer to be turned on and off continuously, which not only results in a certain error in the moisture determination result, but also involves the process of releasing and evacuating the vacuum when the freeze dryer is turned on and off, which affects the entire freeze-drying process and also causes errors in the judgment of the sublimation drying end point; the plate temperature method is applicable to samples with temperature sensors, but if the built-in temperature sensor of the freeze dryer is located on the plate layer, it can also replace the temperature sensor. However, the two built-in temperature sensors of Virtis Advantage2.0 are located in the center of the cavity on both sides of the front box and the rear box, and the temperature sensor located in the front box is close to the freeze dryer door and is also affected by heat conduction and heat radiation. In this article, the cavity temperature is the average value of the two built-in temperature sensors, which is the ambient temperature inside the freeze dryer, and this temperature is higher than the sample temperature; the water line observation method requires that the freeze dryer must have an observation window, the freeze drying mold must be transparent, and the colors of the raw material liquid ice state and the preparation freeze-dried state must be distinct, that is, the sublimation line is obvious and easy to observe. Even if these requirements are met, misjudgment may occur. Therefore, when the sublimation line completely reaches the bottom of the bottle and then disappears, it needs to be maintained for a period of time to ensure complete sublimation.

[0154] Table 10 Method for determining the end point of sublimation drying

[0155]

[0156] 2.5.3 Freeze-drying process evaluation indicators

[0157] (1) Freeze-drying rate

[0158] Due to different set temperatures, the freeze-drying time varies to a certain extent, so the freeze-drying rate formula 1-6 is used to determine the rate of the freeze-drying process.

[0159] Freeze-drying rate formula:

[0160]

[0161] Wherein, F represents the freeze-drying rate (% / h); M 1 Indicates the weight of the raw material solution before freeze-drying (g); M 2 represents the weight of the fast-dissolving tablet after freeze-drying (g); T represents the total freeze-drying time (h).

[0162] (2) Appearance rating

[0163] Score the freeze-dried instant dissolving tablets according to the appearance scoring table under 2.4.1(2).

[0164] (3)Moisture

[0165] According to the first method (Fischer method) under item 0832 of the general chapter of Part III of the 2020 edition of the "Chinese Pharmacopoeia", take an appropriate amount of freeze-dried instant tablets and grind them thoroughly in a mortar. After accurately weighing the weight, quickly place it in the potentiometric titrator and record the measurement results.

[0166] 2.5.4 Establishment of freeze-drying curve

[0167] (1) Pre-freezing stage

[0168] Set the plate temperature to -50℃, -40℃, and -30℃, record the time required to drop to this temperature, and record the cavity temperature inside the instrument at this time, and use the cavity temperature to represent the sample temperature. In the pre-freezing stage, the sample is completely frozen from liquid to ice crystals. To ensure complete freezing, the sample temperature generally needs to be lowered to 10-15℃ below the eutectic point. After the freeze dryer drops to the specified temperature, maintain this temperature for 1h.

[0169] (2) Sublimation drying stage

[0170] The sublimation drying stage is to remove the free water in the material, which mainly exists in the form of ice crystals. After the material is completely frozen into ice crystals, the vacuum degree decreases, and the sublimation drying stage is carried out at this time. The sublimation end point is taken as the end time of sublimation drying. According to Table 11, the sublimation drying time under different sublimation drying curves is examined, and the freeze-drying rate, appearance score, and moisture content are examined.

[0171] Table 11 Sublimation drying stage curve

[0172]

[0173] (3) Drying stage

[0174] The analytical drying stage can remove adsorbed water and bound water in the material. This temperature is usually between 20 and 50°C. Set the plate temperature to 20°C, 25°C, and 30°C. Record the time required for the sublimation drying end temperature to rise from 0°C to this temperature. Also record the cavity temperature inside the instrument at this time. Maintain this temperature for 1 hour and inspect the appearance score and moisture content.

[0175] 3 Experimental results

[0176] 3.1 Clarifying process results

[0177] 3.1.1 Measurement wavelength

[0178] The full wavelength scan results are as follows Figure 3As shown, the transmittance of Platycodon grandiflorum extract solutions diluted 20 times, 30 times and 40 times gradually increased when the wavelength was lower than 660nm, and the transmittance tended to be stable when the measured wavelength exceeded 660nm. Therefore, it was determined that the optimal wavelength for measuring the transmittance of Platycodon grandiflorum extract was 660nm.

[0179] 3.1.2 Results of screening of clarifier types

[0180] The screening results of clarifier types are as follows Figure 4 As shown, pectinase has the best clarification effect at the same concentration. Pectinase precipitates more pectin and macromolecular substances in the extract, making the extract clear. When the pectinase concentration is greater than 1.5g / L, the transmittance exceeds 90%. Therefore, pectinase is determined to be the most suitable clarifier for clarifying Platycodon grandiflorum decoction extract.

[0181] 3.1.3 Results of investigation on pectinase dosage

[0182] The results of the investigation on the dosage of pectinase are as follows Figure 5 As shown, the permeability reached the maximum when the pectinase concentration was 3.0 g / L, and the permeability tended to be stable when the concentration was greater than 3.0 g / L. Therefore, it was determined that the optimal concentration of pectinase added to clarify the Platycodon grandiflorum decoction extract was 3.0 g / L.

[0183] 3.1.4 Clarification time results

[0184] Clarify the time results as Figure 6 As shown in the figure, the permeability reaches the maximum after standing for 8 hours, and the permeability tends to be stable after the time is greater than 8 hours. Therefore, it is determined that the optimal standing time for clarifying the Platycodon decoction extract is 8 hours.

[0185] 3.2 Taste correction study results

[0186] 3.2.1 Establishment of taste standard curve

[0187] (1) Establishment of taste standard curve

[0188] Taste standard curve Figure 7 As shown in the figure, the regression equation of the astringency standard curve is Y=1.2189X+0.9806, R 2 =0.9806; the regression equation of the bitterness standard curve is Y=0.0098X+0.7046, R 2 =0.9767; the regression equation of the sweetness standard curve is Y=0.0118X+0.8297, R 2 =0.9679, indicating that tannic acid has a good linear relationship with the corresponding taste score of VAS in the concentration range of 0.125mg / mL to 8.0mg / mL, quinine sulfate has a good linear relationship with the corresponding taste score in the concentration range of 15.625ng / mL to 1000ng / mL, and sucrose has a good linear relationship with the corresponding taste score of VAS in the concentration range of 12.5mg / mL to 800mg / mL.

[0189] (2) Volunteers selected the most suitable taste range

[0190] The results of volunteers' selection of the most suitable taste are shown in Table 12. The optimum range of VAS scores for astringency is 0.41-4.06; the optimum range of VAS scores for bitterness is 0.47-5.03; and the optimum range of VAS scores for sweetness is 2.75-7.47. These results are used as the target of subsequent taste correction tests.

[0191] Table 12 The most suitable taste results ( n=16)

[0192]

[0193] 3.2.2 Results of the taste analysis of Platycodon grandiflorum decoction

[0194] The results of the taste analysis of Platycodon grandiflorum decoction extract are as follows Figure 8 As shown in the figure, the extract of Platycodon decoction has a high astringency score, a medium bitterness score and a sweetness score. For a long time, Chinese herbal extracts have been "good medicines taste bitter". As a Chinese herbal extract, Platycodon decoction also has the problem of unpleasant taste. However, the licorice contained in Platycodon decoction has a certain sweetness, and licorice not only does not reduce the efficacy of the whole prescription, but also has certain pharmacological effects. It is a safe and palatable masking agent. Therefore, the subsequent taste correction test is mainly to reduce astringency and supplemented by reducing bitterness.

[0195] 3.2.3 Results of the screening of flavoring agents

[0196] The VAS scores of taste of different types of cyclodextrins are shown in Table 13. Although the astringency and bitterness of Platycodon decoction extract are significantly reduced when the dosage of γ-CD is greater than 3.0%, the sourness of Platycodon decoction extract is highlighted. The sourness is almost negligible in the oral test of Platycodon decoction extract and other types of cyclodextrin inclusion. Therefore, γ-CD is not considered as a flavoring agent. The flavoring effect of α-CD is not good. Therefore, HP-β-CD, which can significantly reduce astringency and bitterness and increase sweetness, is selected as a flavoring agent.

[0197] Table 13 Flavor corrective type screening results ( n=16)

[0198]

[0199] 3.2.4 Results of the investigation on the dosage of hydroxypropyl beta-cyclodextrin

[0200] The VAS scoring results of various taste sensations of HP-β-CD at different concentrations are shown in Table 14. When the HP-β-CD dosage is greater than 2.0%, the astringency and bitterness of the Platycodon decoction extract have been significantly reduced, and each taste sensation meets the volunteers' selection of the most suitable taste range; when the HP-β-CD dosage is 3.0%, the astringency and bitterness are better masked, and the sweetness is also improved; when the HP-β-CD dosage is greater than 3.0%, the VAS scores of astringency, bitterness and sweetness all change slightly, so the HP-β-CD dosage of 3.0% is selected, and the VAS scores of each taste sensation meet the volunteers' selection of the most suitable taste range.

[0201] Table 14 Results of HP-β-CD dosage investigation ( n=16)

[0202]

[0203] In order to evaluate whether the Platycodon decoction freeze-dried instant tablets can meet the expected goals, their taste, appearance, disintegration time, etc. were investigated and scored. For Chinese medicine preparations, although the flavor correction process of Platycodon decoction extract was studied in the previous experiments, it is still necessary to pay attention to the taste changes during the prescription screening and freeze-drying process, and the freeze-dried adhesive xanthan gum selected in the excipient screening also has a certain flavor correction function. Therefore, the taste score was added to the subsequent excipient dosage investigation and prescription optimization. The xanthan gum flavor correction acts on the taste part of Platycodon decoction, and the fluffiness of the taste and the appearance are related to the parameters of the freeze-drying process and the amount of excipients. The disintegration time of Platycodon decoction freeze-dried instant tablets is planned to be no more than 1min, and they can usually be completely disintegrated within a few seconds. Too little or too much adhesive will increase the disintegration time, which is related to the properties of the Platycodon decoction extract itself and the solubility of the adhesive in the Platycodon decoction extract. Therefore, the comprehensive score of taste, appearance, and disintegration time is used as the evaluation index when evaluating Platycodon decoction freeze-dried instant tablets.

[0204] 3.3 Screening results of auxiliary materials

[0205] 3.3.1 Screening results of freeze-dried matrix agents

[0206] The appearance scores and disintegration time test results of different types of freeze-dried skeleton agents are shown in Table 15. The effects of maltodextrin and sucrose as skeleton agents are not as good as mannitol. The freeze-dried tablets produced are poor in appearance and tend to collapse after being placed for a period of time. The melting of the tablets is obvious and the formability is poor. Therefore, maltodextrin and sucrose are not considered as freeze-dried skeleton agents. Mannitol has good solubility and is easy to disperse. The properties of the freeze-dried tablets produced are stable. Therefore, mannitol is selected as the freeze-dried skeleton agent based on the appearance score and disintegration time.

[0207] Table 15 Appearance score and disintegration time measurement results ( n=3)

[0208]

[0209] 3.2.2 Freeze-dried adhesive screening results

[0210] The appearance scores and disintegration time test results of different types of freeze-dried adhesives are shown in Table 16. The combination of freeze-dried skeleton agents and adhesives can compensate for the degree of freeze-dried tablets to a certain extent. HPMC K4M and pullulan have poor effects on this experiment. The swelling time of gelatin in the Platycodon decoction extract is relatively long. When the dosage is large, uneven dispersion is prone to occur, resulting in a hard core inside the freeze-dried tablet, which leads to a long disintegration time during the disintegration process. Xanthan gum can achieve the ideal effect at a small dosage. It can effectively prevent the freeze-dried tablets from breaking when used in combination with mannitol, and the tablets have a certain hardness and are easy to demold. Therefore, xanthan gum was selected as the freeze-dried adhesive based on the appearance score and disintegration time.

[0211] Table 16 Appearance score and disintegration time measurement results ( n=3)

[0212]

[0213] 3.4 Results of investigation on auxiliary material dosage

[0214] 3.4.1 Mannitol dosage investigation The taste score, appearance score and disintegration time score showed a trend of first increasing and then decreasing with the increase of mannitol dosage. The mannitol dosage range was 2.0% to 8.0%, indicating that within this dosage range and with a fixed xanthan gum dosage, the performance of the freeze-dried tablets was less affected.

[0215] This experiment uses taste score, appearance score and disintegration time score as evaluation indicators, and uses subjective and objective weighted method to conduct comprehensive scoring. The results are as follows:

[0216] (1) Determine the subjective weight using the AHP method: Based on the importance of each indicator, a pairwise comparison priority judgment matrix (see Table 17) is constructed to calculate the subjective weight coefficient of each indicator (see Table 18). Through the consistency test of the single-level judgment matrix, it can be seen that the consistency ratio factor CR = CI / RI = 0.00 < 0.10, which means that the constructed priority judgment matrix has satisfactory consistency and the calculated subjective weight coefficient is valid.

[0217] Table 17 Pairwise comparison priority judgment matrix

[0218]

[0219] Table 18 Weight calculation results

[0220]

[0221] (2) CRITIC method to determine objective weights: The experimental data are dimensionlessly processed (see Table 19) to calculate the indicators Sj, Rj, Cj and the objective weight WCRITIC. The results are shown in Table 19.

[0222] Table 19 Normalization results of each indicator data

[0223]

[0224] Table 20 Weight coefficients of each indicator

[0225]

[0226] (3) AHP-CRITIC subjective and objective weighting method to determine the comprehensive weight: The comprehensive weight was calculated using the subjective and objective weighting method. The results are shown in Table 21.

[0227] Table 21 Mixed weight coefficients of various indicators

[0228]

[0229] (4) Calculation of comprehensive scores of multiple indicators in the experimental scheme: The scores of each indicator in the experiment were normalized (see Table 22), and the comprehensive scores of each prescription were calculated (see Table 23). The results show that as the amount of mannitol increases, the comprehensive score first increases and then decreases. The data were analyzed by variance analysis using SPSS26.0 software, and the P value was 0.00 < 0.0001, indicating that the amount of xanthan gum has a significant effect on the comprehensive performance of the freeze-dried tablets.

[0230] Table 22 Normalization results of each indicator score

[0231]

[0232]

[0233] Table 23 Comprehensive scoring results

[0234]

[0235] 3.4.2 Investigation of xanthan gum dosage

[0236] The taste score, appearance score and disintegration time score decrease with the increase of xanthan gum dosage within the range of 0.05% to 0.25%. When the concentration reaches 0.25%, the preparation is too viscous and difficult to disperse evenly. Therefore, the concentration of xanthan gum should not exceed 0.25%.

[0237] This experiment uses taste score, appearance score and disintegration time score as evaluation indicators, and uses subjective and objective weighted method to conduct comprehensive scoring. The results are as follows:

[0238] (1) Determination of subjective weights using the AHP method: The subjective weight coefficient is the same as that in 3.4.1(1).

[0239] (2) CRITIC method to determine objective weights: The experimental data are dimensionless (see Table 24), and the indicators Sj, Rj, Cj and objective weights W are calculated. CRITIC , the results are shown in Table 25.

[0240] Table 24 Normalization results of each indicator data

[0241]

[0242] Table 25 Weight coefficients of each indicator

[0243]

[0244] (3) AHP-CRITIC subjective and objective weighting method to determine the comprehensive weight: The comprehensive weight was calculated using the subjective and objective weighting method. The results are shown in Table 26.

[0245] Table 26 Mixed weight coefficients of various indicators

[0246]

[0247] (4) Calculation of comprehensive scores of multiple indicators in the experimental scheme: The scores of each indicator in the experiment were normalized (see Table 27), and the comprehensive scores of each prescription were calculated (see Table 28). The results show that as the amount of xanthan gum increases, the comprehensive score first increases and then decreases. The data were analyzed by variance analysis using SPSS26.0 software, and the P value was 0.00 < 0.0001, indicating that the amount of xanthan gum has a significant effect on the comprehensive performance of the freeze-dried tablets.

[0248] Table 27 Normalization results of each indicator score

[0249]

[0250] Table 28 Comprehensive scoring results

[0251]

[0252] 3.5 Prescription Optimization Results

[0253] 3.5.1 Comprehensive scoring results

[0254] This experiment uses taste score, appearance score and disintegration time score as evaluation indicators, and uses subjective and objective weighted method to conduct comprehensive scoring. The results are as follows:

[0255] (1) Determination of subjective weights using the AHP method: The subjective weight coefficient is the same as that in 3.4.1(1).

[0256] (2) CRITIC method to determine objective weights: The experimental data are dimensionless (see Table 29), and the indicators Sj, Rj, Cj and objective weights W are calculated. CRITIC , the results are shown in Table 30.

[0257] Table 29 Normalization results of each indicator data

[0258]

[0259] Table 30 Weight coefficients of each indicator

[0260]

[0261]

[0262] (3) AHP-CRITIC subjective and objective weighting method to determine the comprehensive weight: The comprehensive weight was calculated using the subjective and objective weighting method. The results are shown in Table 31.

[0263] Table 31 Mixed weight coefficients of various indicators

[0264]

[0265] (4) Calculation of comprehensive scores of multiple indicators in the experimental plan: The scores of each indicator of the experiment were normalized (see 32), and the scores of each indicator were calculated (see Table 33). Finally, the comprehensive scores of each prescription in the central composite design-response surface method experimental design were obtained. The results are shown in Table 34.

[0266] Table 32 Normalization results of each indicator score

[0267]

[0268] Table 33 Scoring results of each indicator

[0269]

[0270] Table 34 Central point design-response surface methodology experimental design and comprehensive scoring results

[0271]

[0272]

[0273] 3.5.2 Data model fitting

[0274] The data were imported into Design-Expert 13.0 software. The comprehensive score of taste, appearance and disintegration time was used as the dependent variable, and the dosage of mannitol (X1) and xanthan gum (X2) in the prescription was used as the independent variable to perform multivariate quadratic regression fitting to obtain the data model. The correlation coefficient (R 2 ) and confidence (P) were used to evaluate the model fitting ability, and the mathematical model was obtained by analysis as follows:

[0275] Z=95.50+3.18X1+1.45X2-0.93X1X2-5.82X1 2 -16.48X2 2 , R 2 =0.9762, P<0.0001.

[0276] By R 2 The results of the P value show that the model fits well. According to the mathematical model, a three-dimensional effect diagram and a contour diagram are drawn, as shown in Fig. 9 shown.

[0277] 3.5.3 Prescription Optimization and Verification

[0278] According to the above fitting equation and three-dimensional effect diagram, it can be seen that with the increase of the dosage of mannitol and xanthan gum, the comprehensive score shows a trend of first increasing and then decreasing. With the maximum comprehensive score of taste score, appearance score and disintegration time score as the goal, a representative prescription was screened and 3 copies were prepared in parallel. The taste score, appearance score and disintegration time score were measured, and the comprehensive score was calculated using the subjective and objective weight coefficients of 3.5.1 (3). The results are shown in Table 35. The experimental results show that the deviation of the optimized formulation prescription is less than 5%, indicating that the model predicts well. The freeze-drying process of this optimized prescription will be investigated later.

[0279] Table 35 Prescription optimization and verification results

[0280]

[0281] Note: Deviation = (predicted value - actual value) / predicted value * 100%

[0282] 3.6 Freeze-drying process results

[0283] 3.6.1 Results of eutectic point and eutectic point determination

[0284] The DSC scanning curve of the optimized formulation is shown in Figure 10, and the results are as follows: the eutectic point and the eutectic point have only one mutation or peak, the eutectic point is -11.03°C and the eutectic point is -0.51°C; the subsequent freeze-drying process is based on this temperature, and the general pre-freezing temperature is 10 to 20°C lower than the eutectic point, and the sublimation temperature in the drying process should generally be controlled below the eutectic point temperature, which is lower than the product collapse temperature.

[0285] 3.6.2 Establishment of freeze-drying curve

[0286] (1) Pre-freezing stage

[0287] The cooling curve of the freeze dryer in the pre-freezing stage is shown in Figure 11, and the results are as follows: the plate layer temperature reaches stability in 1 hour, but the cavity temperature is affected by conduction, and the temperature stabilization time is later than the plate layer temperature stabilization time, but it reaches stability in 2 hours; among them, only when the plate layer temperature is set at -50℃, the cavity temperature can be lower than the eutectic point by 10 to 15℃, that is, the material can be completely frozen at this temperature, and the pre-freezing time is 3 hours.

[0288] (2) Sublimation drying stage

[0289] The results of various indicators in the sublimation drying stage are shown in Table 36. The results are as follows: In the sublimation drying stage, the moisture in the material is transformed from ice into water vapor and sublimates and escapes. Because the sublimation process is an endothermic process, the plate temperature needs to be gradually increased. Curves 1 and 2 are isocratic heating. Although the decrease in vacuum can also make the moisture sublimate, it takes too long and the sublimation rate is low. Therefore, a gradient heating method is selected for sublimation drying. At the same time, if the plate temperature rises too quickly, the sublimation speed of the moisture in the material cannot keep up with the temperature rise speed, which will melt the frozen ice crystals and cause the "boiling" state of the bottle washing, such as curve 3. It is impossible to observe the sublimation line and determine the corresponding indicator results; therefore, a gradient heating method of heating every 10℃ for 20min and maintaining 40min is selected for sublimation drying, and the sublimation drying time is 5h.

[0290] Table 36 Sublimation drying stage measurement results ( n=6)

[0291]

[0292] Note: “—” indicates that no relevant data were measured under this experimental condition.

[0293] (3) Drying stage

[0294] The measurement results of various indicators in the analytical drying stage are shown in Table 37 and the freeze dryer temperature rise curve is shown in Figure 12. The higher the temperature, the lower the moisture content. Increasing the analytical drying temperature can reduce the moisture content of the material to 2% or even below, but low moisture content may cause tablet cracking. From the results, it can be obtained: Therefore, the analytical drying temperature is 25°C and the analytical drying time is 2h.

[0295] Table 37 Analysis of drying stage measurement results ( n=6)

[0296]

[0297] (4) Freeze-drying curve

[0298] The freeze-drying curve is shown in Figure 13, and the results are as follows:

[0299] The present invention is aimed at the classic prescription Jiegeng Decoction. The traditional decoction is inconvenient to use, so a new dosage form is developed to prepare Jiegeng Decoction freeze-dried instant tablets. Based on the previous research on the extraction process of Jiegeng Decoction by the research group, the Jiegeng Decoction extract is clarified and 3g / L pectinase is used to precipitate for 8 hours; the flavoring agent is screened to be 3.0% HP-β-CD; the freeze-dried skeleton agent is screened to be mannitol and the freeze-dried adhesive is xanthan gum. The single-factor experiment determines that the amount of mannitol is 2.0% to 8.0%, and the amount of xanthan gum is 0.05% to 0.25%; the central point design-response surface method is used to optimize the amount of mannitol and xanthan gum, with the maximum comprehensive score as the goal The screening results showed that the dosage of mannitol was 5.06% and the dosage of xanthan gum was 0.15%. The freeze-drying process was screened for the optimized prescription, and the key temperatures were determined: the eutectic point was -11.03℃, the eutectic point was -0.51℃, and the freeze-drying curve was established: the pre-freezing temperature was -50℃, the pre-freezing time was 3h, and the sublimation drying was carried out by gradient heating, starting from -50℃ to 0℃, with a gradient of 10℃, and each gradient heating was maintained for 20min for 40min, the analytical drying temperature was 25℃, and the analytical drying time was 2h.

[0300] Through the preparation prescription and process research of freeze-dried instant tablets of Jingfang Platycodon Decoction, the obtained preparation has suitable taste, appearance and disintegration time, meets the quality evaluation standards, and the stability test provides a basis for the storage and transportation conditions of the tablets. The relative humidity of the storage conditions should be lower than 69.55%. The preparation has consistency in physicochemical properties and in vivo pharmacokinetics with the standard decoction, showing good application prospects.

[0301] Test Example 2 Determination of index components in the freeze-dried instant tablets of the traditional Chinese medicine Jiegeng decoction of the present invention

[0302] 1. Selection of indicator components

[0303] Combined with the reports on active ingredients related to the clinical efficacy of Jiegeng Decoction and network pharmacology research, liquiritin and glycyrrhizic acid in licorice and platycodon saponin D in platycodon were selected as indicator components in this study, and their contents were determined by the method specified in the 2020 edition of the Chinese Pharmacopoeia.

[0304] (1) Investigation of chromatographic conditions

[0305] According to the content determination method under the licorice item in Volume 1 of the 2020 edition of the "Chinese Pharmacopoeia", the contents of liquiritin and glycyrrhizic acid in the freeze-dried instant tablets of Platycodon decoction were determined. C18 (4.6×250 mm, 5 μm) chromatographic column, mobile phase 0.05% phosphoric acid solution (A)-acetonitrile (B), detection wavelength 237 nm, gradient elution as shown in Table 38, flow rate 0.7 mL / min, column temperature 30°C, injection volume 10 μL. Agilent (1260 Infinity) chromatograph was used for analysis.

[0306] Table 38 HPLC-UV licorice content determination gradient elution program

[0307]

[0308] 3. Determination of Platycodon saponin D content

[0309] Take an appropriate amount of instant tablets and grind them thoroughly in a mortar, accurately weigh 2.00g and place it in a stoppered conical flask, accurately add 50mL of 50% methanol, weigh the weight, ultrasonically treat for 30 minutes, cool, weigh again, make up the lost weight with 50% methanol, shake well, and filter; accurately measure 25mL of the filtrate, evaporate to dryness, add 20mL of water to the residue, dissolve it with slight heat, shake and extract it 3 times with n-butanol saturated with water, 20mL each time, combine the n-butanol solution, wash with 50mL of ammonia test solution, discard the ammonia solution, wash with 50mL of water saturated with n-butanol, discard the water solution, recover the solvent with the n-butanol solution until dry, add an appropriate amount of methanol to the residue to dissolve it, transfer it to a 5mL volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate; prepare 3 copies in parallel with the same method, and determine and calculate the content of platycodon saponin D in the instant tablets according to the following method.

[0310] Determination method of platycodon saponin D content

[0311] The content of platycodon saponin D in the samples of Platycodon decoction freeze-dried instant tablets was determined by the content determination method under Platycodon grandiflorum in the 2020 edition of the Chinese Pharmacopoeia. C18 (4.6×250 mm, 5 μm) chromatographic column, mobile phase acetonitrile: water (25:75), isocratic elution for 50 min, flow rate 0.7 mL / min, column temperature 30°C, injection volume 10 μL. Analyzed using Elite (D3200) chromatograph, evaporative light scattering detector (ELSD) parameters: evaporation temperature 110°C, nebulization temperature 50°C.

[0312] The transfer rates of the index components are shown in Table 39. The average transfer rates of the index components liquiritin, glycyrrhizic acid, and platycodon saponin D from the pieces to extract A were 26.67%, 23.06%, and 59.17%, respectively, and the RSDs were 2.22%, 4.42%, and 2.58%, respectively, indicating that the decoction and extraction process of Platycodon decoction had a greater influence on the transfer rates of the index components; the transfer rates of the index components from extract A to extract B, and then to the freeze-dried instant tablets were all greater than 95%, and the RSDs were relatively small, indicating that the losses of the main index components during the extract clarification process investigation, the instant tablet preparation process, and the freeze-drying process were small, and the fluctuations were low, further proving that the preparation process of the freeze-dried instant tablets of Platycodon decoction had small differences, and the preparation process was stable and reliable.

[0313] Table 39 Indicative component transfer rate ( n=6)

[0314]

[0315]

[0316] The established HPLC-UV method can meet the requirements of liquiritin and glycyrrhizic acid content determination, and the HPLC-ELSD method can meet the requirements of platycodon saponin D content determination. It has good specificity and high stability and meets the analytical requirements. The content of liquiritin in Platycodon decoction freeze-dried instant tablets is stable between 0.919% and 0.924%, the content of glycyrrhizic acid is stable between 3.116% and 3.136%, and the content of platycodon saponin D is stable between 0.059% and 0.060%; in the decoction pieces ~ extract A ~Extract B (supernatant) after clarification ~Raw material liquid before freeze-drying of preparation ~Lyophilized instant tabletsIn the study on the quantitative transfer relationship, it was found that the index components had little loss during the extract clarification process, instant tablet preparation and lyophilization process, indicating that the whole process of preparing freeze-dried instant tablets of Jingfang Jiegeng Decoction was highly controllable; the taste scores of the VAS score of the freeze-dried instant tablets all met the optimal taste range selected by the volunteers, indicating that the taste of the freeze-dried instant tablets was good, providing a reference for the taste correction of oral solid preparations of traditional Chinese medicine.

[0317] In addition, the present invention conducted an in vivo comparative study on the standard decoction of Jingfang Jiegeng Decoction and the freeze-dried instant tablets of Jingfang Jiegeng Decoction. Through in vivo pharmacokinetic experiments, it was found that the parameters of liquiritin, glycyrrhizic acid, and platycodon saponin D in the plasma of the freeze-dried instant tablets group A and the freeze-dried instant tablets group B had high similarity; compared with the freeze-dried instant tablets group, the AUC of liquiritin and glycyrrhizic acid in the standard decoction administration group were 1.09 times that of the freeze-dried instant tablets group, and the Cmax was 1.11 times that of the freeze-dried instant tablets group; the AUC of platycodon saponin D in the standard decoction administration group was 0.81 times that of the freeze-dried instant tablets group, and the Cmax was 1.09 times that of the freeze-dried instant tablets group, so the plasma drug concentration was consistent.

[0318] On the basis that the preparation meets the expected performance, in order to further clarify the consistency of efficacy between the freeze-dried instant tablets of Jingfang Jiegeng Decoction and the standard decoction, a plasma pharmacokinetic experiment was designed. The pharmacokinetics of freeze-dried orodisintegrating tablets are mostly experimental animals, and there are usually two methods of administration: one is sublingual mucosal administration, when administering, the tongue of the beagle dog needs to be stretched first, the orodisintegrating tablet is placed under the tongue, and the dog's tongue is pressed for 5 minutes without moving, and blood can be drawn after the orodisintegrating tablet is completely dissolved; the other is supralingual administration, when administering, the drug is placed on the tongue of the beagle dog, the dog's mouth is pressed for 5 minutes without moving, and blood can be drawn after the orodisintegrating tablet is completely dissolved. This study developed a fast-dissolving dosage form with the same efficacy as Platycodon decoction. The main way of administration is to dissolve in water, and it can be taken orally without water when necessary. Therefore, the same delivery form as the standard decoction was adopted, that is, dissolving in water before oral administration. SD rats were used as experimental animals, and the concentrations of licorice, glycyrrhizic acid and platycodon saponin D in plasma of the standard decoction, freeze-dried instant tablet group A and freeze-dried instant tablet group B were compared. It was found that the concentrations of licorice, glycyrrhizic acid and platycodon saponin D in plasma of the freeze-dried instant tablets were consistent with those of the standard decoction regardless of the amount of water added. This demonstrated the consistency of the efficacy of the freeze-dried instant tablets of Platycodon decoction and the standard decoction.

[0319] In summary, the present invention provides a research idea for flavor correction for the development of Chinese medicine preparations. Chinese medicine extracts have been said to be "bitter medicines" since ancient times. When developing oral solid preparations, it is necessary to focus on the flavor correction process, use visual analogue scoring (VAS) for evaluation, screen and train volunteers through the three taste levels of astringency, bitterness and sweetness, establish a taste standard curve, select the most suitable taste range, and use the Grubb test method to perform a circular test and eliminate outliers on the data, converting subjective descriptions into objective scores of different tastes, providing ideas for flavor correction research for the development of oral preparations of Chinese medicine. Provide research ideas for the development of preparations of classic prescriptions. According to the current "Drug Registration Management Measures", traditional prescriptions are mainly developed into granules, tablets, capsules, etc., and the extracts need to be heated, concentrated, dried, and water removed during their preparation to facilitate molding. However, the study found that during the long concentration and drying process, the active ingredients were degraded by heat, the solubility of the extract decreased, and the dispersion state of the ingredients after the granules were re-dissolved in water was quite different from that of the decoction. The dissolution and dispersion of the active ingredients in the tablets and capsules prepared by them is even more unclear, which has become a common problem restricting the clinical efficacy of solid preparations of classical prescriptions. The ingredients of Chinese medicine preparations are complex. The freeze-drying technology is used to remove the moisture in the materials at low temperature, and the ingredient retention rate is higher; freeze-drying forms a high-porosity structure with a loose texture. After adding water, it can be quickly dissolved and restore the original dispersed phase of the decoction, and the solubility is significantly better than that of conventional solid preparations; by comparing the pharmacokinetic evaluation of the standard decoction of Platycodon decoction and the freeze-dried instant tablets, it is confirmed that the Platycodon decoction freeze-dried instant tablets have the same efficacy as the standard decoction of Platycodon decoction; the preparation process of Platycodon decoction freeze-dried instant tablets is simple, the water content is low, the stability is high, the use is convenient, the patient compliance is high, the dosage is accurate, and it is convenient to transport and carry, providing research ideas for the development of preparations of classic prescriptions. Innovation in the application of dual-purpose medicinal and edible varieties: Platycodon and licorice in Platycodon decoction are included in the catalog of dual-purpose medicinal and edible varieties released by the National Health Commission, and freeze-dried instant tablets have been seen. Based on the concept of strengthening the development of health foods proposed in the "Outline of the Strategic Planning for the Development of Traditional Chinese Medicine (2016-2030)", in accordance with the requirements of the "Health Food Registration and Filing Management Measures", on the basis of the functional value of the classical prescription Platycodon decoction, from the perspective of public demand, research on giving it food attributes can enrich the application forms of this dual-purpose medicinal and edible variety.

Claims

1. A method for preparing Platycodon decoction freeze-dried instant tablets, characterized in that: It includes the following steps: a. Weigh the raw materials in the weight ratio: 4-6 parts of Platycodon grandiflorum, 8-12 parts of Licorice root; b. Extraction: Soak in water, decoct, filter and obtain the filtrate; c. Clarification: add a clarifier to the extract of step b, let it stand, filter, and obtain a filtrate; the clarifier is pectinase, the dosage is 3g / L, and the standing time is 8h; d. Freeze-drying: After concentrating the filtrate in step c, add a flavoring agent, a freeze-dried skeleton agent, and a freeze-dried binder, wherein the amounts of the flavoring agent, the freeze-dried skeleton agent, and the freeze-dried binder are calculated based on the clarified filtrate in step c, respectively: Flavor corrective 3%, freeze-dried skeleton 5.06%, freeze-dried adhesive 0.15%; Freeze-drying temperature: eutectic point is -11.03°C, eutectic point is -0.51°C; The flavor corrector is HP-β-CD; the freeze-dried skeleton agent is mannitol; the freeze-dried adhesive is xanthan gum; e. Tableting; The freeze-drying conditions described in step d are: pre-freezing temperature -50°C, pre-freezing time 3 h, sublimation drying using a gradient heating method, starting from -50°C to 0°C, a gradient of 10°C, each gradient heating for 20 min and maintaining for 40 min, analytical drying temperature 25°C, analytical drying time 2 h.

2. The preparation method according to claim 1, characterized in that: The ratio of the raw materials described in step a is: 5 parts of Platycodon grandiflorum and 10 parts of Licorice root; the concentration of the Platycodon grandiflorum decoction extract described in step b is 0.5 g / mL.

3. The Jiegeng decoction freeze-dried instant tablets prepared by the method for preparing Jiegeng decoction freeze-dried instant tablets according to claim 1 or 2, characterized in that: It contains 0.919%-0.924% liquiritin, 3.116%-3.136% glycyrrhizic acid, and 0.059%-0.060% platycoside D.

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