Isatis root granules and a preparation method thereof

Abstract

The application provides a radix isatidis granule and a preparation method thereof, which is prepared from the following components in parts by weight: radix isatidis 700 parts, sucrose 170-190 parts and dextrin 50-70 parts. The content of active ingredient (R,S)-deoxyschizandrin, the total content of uridine, guanosine and adenosine, and the content of indirubin in the prepared radix isatidis granule are significantly improved, and the quality is stable after being accelerated for 6 months.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine preparations and relates to a Banlangen granule and its preparation method. Background Technology

[0002] Isatis root granules are formulated with Isatis root and have the effects of clearing heat and detoxifying (antiviral) and cooling blood; they are used to treat fever, rashes, wind-heat cold, sore throat, epidemic encephalitis B, hepatitis, and mumps.

[0003] The 2020 edition of the Chinese Pharmacopoeia describes the preparation method of Banlangen granules as follows:

[0004]

Preparation

[0005] Alternatively, ethanol can be recovered and concentrated to a clear paste with a relative density of about 1.25 (60-65℃), dried, and the dried paste can be mixed with an appropriate amount of sweetener to form granules, dried, and made into 500g [specification (5)]; or ethanol can be recovered and concentrated to a clear paste with a relative density of about 1.10 (50℃), spray dried, and the dried extract powder can be mixed with an appropriate amount of maltodextrin, dextrin and stevioside, mixed evenly, formed into granules, dried, and made into 360g [specification (6)]; or ethanol can be recovered and concentrated to a relative density of 1.32-1.35 (60℃), dried, pulverized, and an appropriate amount of starch and humectant can be added, mixed evenly, formed into granules, dried, and made into 200g [specification (7)], which is the final product.

[0006] However, there are still significant problems with product quality. The Changzhou Food and Drug Supervision and Inspection Center in Jiangsu Province sampled 18 batches of Banlangen granules from different manufacturers on the market for quality analysis. The study expanded to include content determination and testing for heavy metals and harmful elements. It was found that due to outdated standards, there were significant differences in quality between different manufacturers and different batches.

[0007] Therefore, we still need to research and develop the formulation and production process of Banlangen granules, improve quality standards, and ensure consistent quality between and within batches. Summary of the Invention

[0008] The inventors of this invention have long been dedicated to the research of Isatis root granules. During their research, they unexpectedly discovered that using a specific extraction method significantly increases the content of active ingredients such as (R,S)-gayopicrin, uridine, guanosine, total adenosine, and indirubin in the Isatis root granules, resulting in a significantly enhanced antiviral effect. After being placed under accelerated conditions for 6 months, the granules exhibit good stability.

[0009] First, the present invention provides a granule of Isatis indigotica root, which is prepared from the following components in parts by weight:

[0010] Components Number of weights Isatis root 700 sucrose 170-190 dextrin 50-70

[0011] The dextrin is composed of cyclodextrin and maltodextrin in a weight ratio of 1:2; the cyclodextrin is preferably sodium sulfobutylbetacyclodextrin.

[0012] Furthermore, an Isatis root granule is prepared from the following components in parts by weight:

[0013] Components Number of weights Isatis root 700 sucrose 175-185 dextrin 55-65

[0014] Furthermore, a type of Isatis root granule is prepared from the following components in parts by weight:

[0015] Components Number of weights Isatis root 700 sucrose 180 dextrin 60

[0016] Furthermore, each bag of the Isatis root granules contains 5-10g, preferably 5g or 10g, and more preferably 10g.

[0017] Furthermore, the present invention also provides a method for preparing the above-mentioned Isatis root granules, comprising the following steps:

[0018] (1) Preparation of Isatis root extract

[0019] ① Washing and soaking: Wash the Isatis root with clean water 1-3 times, using 2-3 times the amount of water each time, based on the weight of the Isatis root. After washing, add the Isatis root to a 1%-1.4% potassium permanganate solution, with the weight of the potassium permanganate solution being 3-4 times the weight of the Isatis root. Control the temperature at 0-5℃ and stir for 2-3 hours. Then raise the temperature to 15-20℃ and stir for 3-5 hours.

[0020] ② Cutting and drying: Cut the Isatis root into segments ≤7cm and dry under reduced pressure until the moisture content is below 10%;

[0021] ③ Extraction: Isatis root is decocted twice with water. For the first decoction, add 8 times the amount of water and 60%-70% sodium sulfobutyl betacyclodextrin, decoct for 2 hours, and pour off the decoction. For the second decoction, add 6 times the amount of water and the remaining sodium sulfobutyl betacyclodextrin, decoct for 1 hour, pour off the decoction, filter the decoction, and combine the filtrates.

[0022] ④ Concentration and alcohol precipitation: Concentrate the filtrate to a clear extract with a relative density of 1.18-1.22, add ethanol to make the alcohol content reach 60%-65%, let stand for more than 24 hours to precipitate, take the supernatant, recover the ethanol and concentrate to a density of 1.25-1.28 to obtain the Isatis root extract.

[0023] (2) Granulation and drying

[0024] ①Preparation: Grind sucrose into powder, pass it through an 80-mesh sieve, heat the Isatis root extract to 50-70℃ and maintain it, then set aside.

[0025] ② Granulation: After mixing sucrose and dextrin for 10 minutes, slowly add Isatis root extract over 5-8 minutes while stirring at 180-200 rpm and cutting at 1500-1600 rpm to granulate.

[0026] ③ Drying and packaging: The prepared Isatis root granules are dried and packaged.

[0027] Furthermore, in step (1), a 1.2% potassium permanganate solution is preferred.

[0028] Furthermore, in step (1), the potassium permanganate solution is preferably 3.5 times the weight of Isatis root.

[0029] Furthermore, in step (1), it is preferable to control the temperature at 2-5℃ and stir for 2.5h; then raise the temperature to 15-18℃ and stir for 4h.

[0030] Furthermore, in step (1), it is preferable to add 65% sodium sulfobutyl betacyclodextrin.

[0031] Further, in step (2), the preferred Isatis root extract is heated to 60°C and maintained.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] (1) The applicant found that when Isatis root was pretreated by soaking before extraction, that is, by soaking Isatis root in a potassium permanganate solution of a specific concentration, the content of active ingredient (R,S)-Gouyichun, uridine, guanosine and adenosine was significantly increased. In particular, the content of Isatis root-specific antiviral component (R,S)-Gouyichun reached more than 5mg / bag, which significantly improved the antiviral effect of Isatis root granules.

[0034] (2) The inventors have discovered that adding sodium sulfobutyl betacyclodextrin during the decoction of Isatis indigotica can efficiently extract the total amount of the effective components in Isatis indigotica, uridine, guanosine and adenosine, especially the content of indigotica reaching more than 270 μg / bag, which further significantly enhances the antiviral effect of Isatis indigotica granules.

[0035] (3) Since the extract of Isatis indigotica is an extract, it is easy to agglomerate into lumps during the preparation process, resulting in large differences in the amount of content. This invention optimizes the preparation method, especially the addition temperature, addition time, stirring speed and cutting speed of the extract of Isatis indigotica during granulation. The prepared Isatis indigotica granules have uniform content and small differences in content. Detailed Implementation

[0036] This invention discloses a Banlangen granule and its preparation method. Those skilled in the art can refer to the content of this invention and, in conjunction with the relevant principles of pharmaceutical preparations, appropriately modify the process parameters to achieve the same result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included within the scope of this invention. The application of this invention has been described through preferred embodiments, and those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0037] To better understand the invention and not to limit its scope, all figures indicating amounts, percentages, and other numerical values ​​used in this application should, in all cases, be understood to be modified by the word "approximately." Each numerical parameter should at least be considered as obtained based on reported significant figures and through conventional rounding methods.

[0038] The present invention will be further illustrated by the following examples, but the examples do not limit the present invention in any way.

[0039] Content uniformity: The content uniformity was determined according to the method for content uniformity in the General Chapter on Granules, Part III, Chinese Pharmacopoeia 2020 Edition.

[0040] Uric acid, guanosine, and adenosine: determined according to the content detection method in Banlangen granules of the 2020 edition of the Chinese Pharmacopoeia.

[0041] The (R,S)-Gao Yichun detection method is as follows:

[0042] Chromatographic conditions: Column: Shimadzu VP-ODS C 18 (150mm×4.6mm, 5μm); Mobile phase: methanol-0.05% phosphoric acid solution (7:93, v / v); Flow rate: 1.0ml / min; Detection wavelength: 245nm; Column temperature: room temperature; Injection volume: 10μl; The theoretical plate number (R,S)-Gao Yichun Peak should not be less than 4000, and the resolution should be greater than 1.5.

[0043] Preparation of reference solution: Accurately weigh approximately 12.5 mg of (R,S)-Gouyichun reference standard and place it in a 25 ml volumetric flask. Dilute with water to the mark and shake well. Accurately pipette 1 ml into a 10 ml volumetric flask and add water to prepare a (R,S)-Gouyichun reference solution containing 50 μg per ml.

[0044] Preparation of the test solution: Take an appropriate amount of the contents of this product, grind it into a fine powder, take about 0.5g, weigh it accurately, place it in a stoppered conical flask, add 20ml of water accurately, weigh it, heat it in a water bath and shake it to dissolve it, cool it, weigh it again, add water to make up the lost mass, shake it well, filter it, and take the filtrate to obtain the test solution.

[0045] Detection method: Inject the reference solution and the test solution into the liquid chromatograph, record the chromatogram, and calculate the content.

[0046] Indigo red detection method:

[0047] Chromatographic conditions: Column: Agilent C1000 18 (250mm×4.6mm, 5μm); Mobile phase: methanol-water solution (69:31, v / v); Flow rate: 1.0ml / min; Detection wavelength: 293nm; Column temperature: room temperature; Injection volume: 10μl; The theoretical plate number based on the indigo red peak should be no less than 2000, and the resolution should be greater than 1.5.

[0048] Preparation of reference solution: Accurately weigh about 10 mg of indirubin reference standard and place it in a 100 ml volumetric flask. Dilute to the mark with methanol and shake well. Accurately pipette 1 ml into a 10 ml volumetric flask and add methanol to prepare a reference solution containing 10 μg of indirubin per ml.

[0049] Preparation of the test solution: Take an appropriate amount of the contents of this product, grind it into a fine powder, weigh about 10g accurately, place it in a 50ml volumetric flask, add 30ml of methanol, sonicate for 20 minutes, add methanol to the mark, shake well, centrifuge, and take the supernatant to obtain the test solution.

[0050] Detection method: Inject the reference solution and the test solution into the liquid chromatograph, record the chromatogram, and calculate the content.

[0051] Examples 1-5: Isatis root granules

[0052] I. Prescription Composition:

[0053] Prepare Isatis root granules by adding ingredients according to the prescription of 5000 bags, the same applies below.

[0054] Components Example 1 Example 2 Example 3 Example 4 Example 5 Isatis root 70Kg 70Kg 70Kg 70Kg 70Kg sucrose 18Kg 17.7Kg 18.3Kg 17.5Kg 18.5Kg dextrin 6Kg 6.3Kg 5.7Kg 6.5Kg 5.5Kg

[0055] The dextrin is composed of cyclodextrin and maltodextrin in a weight ratio of 1:2; the cyclodextrin is sodium sulfobutylbetacyclodextrin.

[0056] II. Preparation method:

[0057] (1) Preparation of Isatis root extract

[0058] ① Washing and soaking: Wash the Isatis root twice with clean water, using 150 kg of water each time; after washing, add the Isatis root to a 1.2% potassium permanganate solution, with a potassium permanganate solution weighing 240 kg, control the temperature at 2-5℃, and stir for 2.5 h; then raise the temperature to 15-18℃ and stir for 4 h.

[0059] ② Cutting and drying: Cut the Isatis root into segments ≤7cm and dry under reduced pressure until the moisture content is 8.2%;

[0060] ③ Extraction: Isatis root is decocted twice with water. For the first decoction, add 8 times the amount of water and 65% sodium sulfobutyl betacyclodextrin, and decoct for 2 hours. Pour off the decoction. For the second decoction, add 6 times the amount of water and the remaining sodium sulfobutyl betacyclodextrin, and decoct for 1 hour. Pour off the decoction, filter the decoction, and combine the filtrates.

[0061] ④ Concentration and alcohol precipitation: Concentrate the filtrate to a clear extract with a relative density of 1.18-1.22, add ethanol to make the alcohol content reach 60%-65%, let stand for more than 24 hours to precipitate, take the supernatant, recover the ethanol and concentrate to a density of 1.25-1.28 to obtain the Isatis root extract.

[0062] (2) Granulation and drying

[0063] ①Preparation: Grind sucrose into powder, pass through an 80-mesh sieve, heat the Isatis root extract to 60℃ and maintain the temperature, set aside.

[0064] ② Granulation: After mixing sucrose and dextrin for 10 minutes, slowly add Isatis root extract over 5-8 minutes while stirring at 180-200 rpm and cutting at 1500-1600 rpm to granulate.

[0065] ③ Drying and packaging: The prepared Isatis root granules are dried and packaged into 10g bags.

[0066] Isatis root granules were obtained.

[0067] III. Test Results:

[0068] (1) Filling volume difference: as shown in the table below

[0069] (2) Content of active ingredients (R, S) - uridine, guanosine, total adenosine, indirubin, etc.

[0070]

[0071] The dosage of the Isatis root granules prepared by this invention meets the requirements.

[0072] The Isatis root granules prepared by this invention have a (R,S)-Gauzexin content of more than 5 mg / bag, a total amount of uridine, guanosine, and adenosine of more than 15 mg / bag, and indirubin of more than 270 μg / bag, which ensures the antiviral therapeutic effect of Isatis root. Experiments have shown that the therapeutic effect is significantly better than that of commercially available preparations.

[0073] Comparative Example 1: Isatis Root Granules

[0074] I. Prescription Composition:

[0075] Components Comparative Example 1 Isatis root 70Kg sucrose 18Kg dextrin 6Kg

[0076] The dextrin mentioned is maltodextrin.

[0077] II. Preparation method:

[0078] (1) Preparation of Isatis root extract

[0079] ① Washing and soaking: Wash the Isatis root twice with clean water, using 150 kg of water each time;

[0080] ② Cutting and drying: Cut the Isatis root into segments ≤7cm and dry under reduced pressure until the moisture content is 8.5%;

[0081] ③ Extraction: Isatis root is decocted twice with water. For the first decoction, add 8 times the amount of water and decoct for 2 hours, then pour off the decoction. For the second decoction, add 6 times the amount of water and decoct for 1 hour, then pour off the decoction. Filter the decoction and combine the filtrates.

[0082] ④ Concentration and alcohol precipitation: Concentrate the filtrate to a clear extract with a relative density of 1.18-1.22, add ethanol to make the alcohol content reach 60%-65%, let stand for more than 24 hours to precipitate, take the supernatant, recover the ethanol and concentrate to a density of 1.25-1.28 to obtain the Isatis root extract.

[0083] (2) Granulation and drying

[0084] ①Preparation: Grind sucrose into powder, pass through an 80-mesh sieve, heat the Isatis root extract to 60℃ and maintain the temperature, set aside.

[0085] ② Granulation: After mixing sucrose and dextrin for 10 minutes, slowly add Isatis root extract over 5-8 minutes while stirring at 180-200 rpm and cutting at 1500-1600 rpm to granulate.

[0086] ③ Drying and packaging: The prepared Isatis root granules are dried and packaged into 10g bags.

[0087] Isatis root granules were obtained.

[0088] III. Test Results:

[0089] (1) Filling volume difference: as shown in the table below

[0090]

[0091] (2) Content of active ingredients (R, S) - uridine, guanosine, total adenosine, indirubin, etc.

[0092]

[0093] The Isatis root granules prepared according to the pharmacopoeia method in Comparative Example 1 had a (R,S)-Gauzecin content of only 2.47 mg / bag, which was significantly worse than the 5 mg / bag or more in Examples 1-5 of the present invention; the total amount of uridine, guanosine, and adenosine was only 8.81 mg / bag, which was significantly worse than the 15 mg / bag or more in Examples 1-5 of the present invention; and the amount of indirubin was only 180.54 μg / bag, which was significantly worse than the 270 μg / bag or more in Examples 1-5 of the present invention. The antiviral treatment effect was significantly worse than that in Examples 1-5.

[0094] Comparative Examples 2-6: Effects of Soaking Treatment on Isatis Root Granules

[0095] I. Prescription Composition: Same as Example 1

[0096] II. Preparation method: The “① cleaning and soaking” steps in step (1) are shown in the table below. The remaining steps are the same as in Example 1.

[0097]

[0098] III. Test Results:

[0099] (1) Filling volume difference: as shown in the table below

[0100]

[0101] (2) Content of active ingredients (R, S) - uridine, guanosine, total adenosine, indirubin, etc.

[0102]

[0103] The only difference between Comparative Examples 2-6 and Example 1 is the soaking process of Isatis root. The content of their active ingredients is significantly lower than that of Example 1, especially the content of (R,S)-goichun, uridine, guanosine, and total adenosine. The antiviral therapeutic effect is significantly lower than that of Example 1.

[0104] Based on the detection results of active ingredients in Examples 1-5 and Comparative Examples 2-6, before extracting Isatis root, the Isatis root was pretreated by soaking, preferably with a 1.2% potassium permanganate solution, more preferably at a controlled temperature of 2-5℃, and stirred and soaked for 2.5h; then the temperature was raised to 15-18℃ and stirred and soaked for 4h. The total amount of uridine, guanosine and adenosine in the prepared Isatis root granules was significantly increased, and the content of (R,S)-Gauzymin reached more than 5mg / bag.

[0105] Comparative Examples 7-9: Effect of the addition of sodium sulfobutylbetacyclodextrin during the extraction step on Isatis indigotica granules

[0106] I. Prescription Composition: The prescription for Comparative Example 7 is as follows, and the prescriptions for Comparative Examples 8-9 are the same as those for Example 1.

[0107] Components Comparative Example 7 Isatis root 70Kg sucrose 18Kg maltodextrin 6Kg

[0108] II. Preparation method: The “③ Extraction” step in step (1) is shown in the table below. The remaining steps are the same as in Example 1.

[0109]

[0110] III. Test Results:

[0111] (1) Filling volume difference: as shown in the table below

[0112] (2) Content of active ingredients (R, S) - uridine, guanosine, total adenosine, indirubin, etc.

[0113]

[0114] The only difference between Comparative Examples 7-9 and Example 1 is whether sodium sulfobutyl betacyclodextrin is added during the extraction of Isatis indigotica. The content of active ingredients in Comparative Examples 7-9 is significantly worse than that in Example 1, especially the content of indirubin, uridine, guanosine, and total adenosine. According to the experiment, its antiviral therapeutic effect is significantly worse than that in Example 1.

[0115] Based on the detection results of active ingredients in Examples 1-5 and Comparative Examples 7-9, it is preferable to add sodium sulfobutyl betacyclodextrin during the decoction extraction of Isatis indigotica root. More preferably, 65% sodium sulfobutyl betacyclodextrin is added during the first decoction, and the remaining sodium sulfobutyl betacyclodextrin is added during the second decoction extraction. The total amount of uridine, guanosine, and adenosine in the prepared Isatis indigotica root granules is significantly increased, and the content of indirubin reaches more than 270 μg / bag.

[0116] Comparative Example 10-11: Isatis Root Granules

[0117] I. Prescription Composition: Same as Example 1

[0118] II. Preparation method: The steps “① Preparation” and “② Granulation” in step (2) are shown in the table below. The remaining steps are the same as in Example 1.

[0119]

[0120] III. Test Results:

[0121] (1) Filling volume difference: as shown in the table below

[0122] (2) Content of active ingredients (R, S) - uridine, guanosine, total adenosine, indirubin, etc.

[0123] Because the dosage does not meet the requirements, the content of active ingredients will no longer be tested.

[0124] The results of Comparative Examples 10-11 show that the addition temperature, addition time, stirring speed, and cutting speed of Isatis indigotica extract during granulation affect the difference in the amount of clothing applied.

[0125] Therefore, based on the results of the content difference in Examples 1-5 and Comparative Examples 10-11, it is preferable to heat the Isatis root extract to 60°C and maintain it during granulation, slowly add the Isatis root extract over a period of 5-8 minutes, stir at 180-200 rpm, and cutter at 1500-1600 rpm. The content difference of the prepared Isatis root granules meets the requirements.

[0126] The Isatis root granules prepared in Examples 1-5 of this invention showed good stability after being placed under accelerated conditions for 6 months. The contents of active ingredients such as (R, S)-Gouyichun, uridine, guanosine, total adenosine, and indirubin did not change significantly.

Claims

1. A granule of radix isatidis, prepared from the following components by weight: radix isatidis 700 parts, sucrose 170-190 parts, dextrin 50-70 parts. The dextrin is composed of cyclodextrin and malt dextrin in a weight ratio of 1:2; the cyclodextrin is sulfobutyl betadex sodium. The preparation method comprises the following steps: (1) Preparation of radix isatidis extract ① Washing and soaking: wash the radix isatidis with water 1-3 times, with the amount of water being 2-3 times the weight of the radix isatidis each time; after washing, add 1%-1.4% potassium permanganate solution to the radix isatidis, with the weight of the potassium permanganate solution being 3-4 times the weight of the radix isatidis, control the temperature at 0-5℃, and stir for 2-3 hours; increase the temperature to 15-20℃, and stir for 3-5 hours; ② Cutting and drying: cut the radix isatidis into segments ≤7 cm, and dry under reduced pressure until the moisture content is less than 10%; ③ Extraction: decoct the radix isatidis twice with water, add 8 times the amount of water for the first time, add 60%-70% sulfobutyl betadex sodium, decoct for 2 hours, and pour out the decoction; add 6 times the amount of water for the second time, add the remaining sulfobutyl betadex sodium, decoct for 1 hour, pour out the decoction, filter the decoction, and combine the filtrates; ④ Concentration and alcohol precipitation: concentrate the filtrate to a clear extract with a relative density of 1.18-1.22, add ethanol to a content of 60%-65%, stand for 24 hours or more, precipitate, take the supernatant, recover the ethanol, and concentrate to a density of 1.25-1.28 to obtain the radix isatidis extract; (2) Granulation and drying ① Preparation: powder sucrose, pass through an 80-mesh sieve, and heat the radix isatidis extract to 50-70℃ and keep it ready; ② Granulation: mix sucrose and dextrin for 10 minutes, then slowly add the radix isatidis extract, with the addition time being 5-8 minutes, the stirring speed being 180-200 rpm, and the cutter speed being 1500-1600 rpm, to granulate; ③ Drying and packaging: dry and package the prepared radix isatidis granules.

2. The radix isatidis granules according to claim 1, wherein, Prepared from the following components: radix isatidis 700 parts, sucrose 175-185 parts, dextrin 55-65 parts.

3. The radix isatidis granules according to claim 2, wherein, Prepared from the following components: radix isatidis 700 parts, sucrose 180 parts, dextrin 60 parts.

4. The Isatis root granules as described in claim 1, characterized in that, In step (1), 1.2% potassium permanganate solution.

5. The Isatis root granules as described in claim 1, characterized in that, In step (1), the weight of the potassium permanganate solution is 3.5 times the weight of the radix isatidis.

6. The Isatis root granules as described in claim 1, characterized in that, In step (1), control the temperature at 2-5℃, and stir for 2.5 hours; increase the temperature to 15-18℃, and stir for 4 hours.

7. The Isatis root granules as described in claim 1, characterized in that, In step (1), add 65% sulfobutyl betadex sodium.

8. The Isatis root granules as described in claim 1, characterized in that, In step (2), heat the radix isatidis extract to 60℃ and keep it.

9. The Isatis root granules as described in claim 1, characterized in that, 10 g per bag.

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