RU2778292C1 - Granular complex ketone ether of ginkgo biloba and method for its preparation - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: group of inventions relates to the pharmaceutical industry, namely to the granular composition of ginkgo biloba leaf extract, the method for its preparation and its application. A method for obtaining a granular composition from ginkgo biloba leaf extract, includes the following steps: A) mixing and stirring of ginkgo biloba leaf extract, dextrin, sucrose powder, starch, low-substituted hydroxypropyl cellulose and aspartame in accordance with the ratio in the composition and adding at the same time an aqueous solution of ethanol to obtain a soft material; B) obtaining wet granules from soft material, followed by drying of wet granules; and C) sieving and straightening of dried granules, mixing, releasing and packaging into bags to obtain a granular composition from ginkgo biloba leaf extract; where the ratio in the composition in weight parts includes the following: 35-45 parts of ginkgo biloba leaf extract; 450-500 parts of dextrin; 420-450 parts sucrose powder; 10-30 parts starch; 10-30 parts low-substituted hydroxypropyl cellulose; and 5-10 parts aspartame; moreover, extraction to obtain ginkgo biloba leaf extract includes the following stages: a) crushing of ginkgo biloba leaves to obtain crushed ginkgo biloba leaves; b) adding an aqueous ethanol solution to crushed ginkgo biloba leaves, conducting a double extraction by heating with a reverse refrigerator and obtaining the first filtrate by filtering; c) adding water to the filtered residue of the drug remaining after filtration, conducting a single extraction by heating with a reverse refrigerator, and then obtaining a second filtrate by filtration; d) combining and concentrating the second filtrate and the first filtrate into a thick paste, dissolving in purified water, cooling and filtering to obtain a filtrate; e) loading the filtrate into a column with a macroporous resin, sequential elution with 18% aqueous ethanol solution, 30% aqueous ethanol solution and 50% aqueous ethanol solution to obtain 18% ethanol eluent, 30% ethanol eluent and 50% ethanol eluent, respectively, which are the first eluents; in the first eluents, 50% ethanol eluent it is the first eluent with a high concentration of solvent, and the combined solution of 18% ethanol eluent and 30% ethanol eluent is the first eluent with a low concentration of solvent; f) concentration of the first eluent with a low concentration of solvent so that there is no smell of alcohol, loading the first eluent with a low concentration of solvent into a polyamide column, and then elution with ethanol to obtain ethanol eluate, which is the second eluate; g) combining and concentrating the second eluent and the first eluent with a high concentration of solvent until the smell of alcohol disappears; h) extraction of concentrate with cyclohexane, discarding cyclohexane extract, spray drying of concentrated solution or concentration of concentrated solution into a thick paste and vacuum drying of thick paste; grinding of dried products to obtain ginkgo biloba leaf extract; and where the granular composition of ginkgo biloba leaf extract, based on the loading weight of each package of 1 g, satisfies the following conditions: 1) the content of rutin is less than or equal to 1.60 mg/bag; 2) the content of quercetin is less than or equal to 0.160 mg/bag; 3) the content of bilobalide is 1.04-2.08 mg/bag; 4) the content of ginkgolide J is 0.04-0.20 mg/bag; 5) the residual ethanol content is less than or equal to 0.2 mg/bag; 6) the content of biflavonoids is less than or equal to 0.008 mg/bag, and these biflavonoids include amentoflavone, bilobetin and ginkgetin; 7) the content of genistin is 0, and the content of ginkgolide M is 0; 8) the content of terpene lactones is 2.4-4.8 mg/bag; 9) the content of total flavonoids is 14.08-26.40 mg/bag; and 10) the content of total flavonol glycosides is 9.6-14.0 mg/bag. Granular composition of ginkgo biloba leaf extract, providing anti-PAF (platelet activation factor) effect and nerve nutrition obtained by the above method, where the content of total ginkgolic acids in the granular composition of ginkgo biloba leaf extract is less than or equal to 2 mcg/bag; where total ginkgolic acids in the granular composition of ginkgo biloba leaf extract is calculated based on the total content of ginkgolic acid C13:0, ginkgolic acid C15:1 and ginkgolic acid C17:1. A granular composition from ginkgo biloba leaf extract, providing an anti-PAF effect and nerve nutrition, obtained by the above method, where the ratio of the areas of chromatographic peaks of quercetin to kaempferide in the granular composition of ginkgo biloba leaf extract is 0.8-1.2, and the ratio of the areas of chromatographic peaks of isoramnetin to quercetin in the granular composition of ginkgo biloba leaf extract is greater than 0.15. The above method can be used to obtain a granular composition from ginkgo biloba leaf extract, which provides an anti-PAF effect and nerve nutrition.

EFFECT: above-described granular composition has a high therapeutic activity, providing an anti-PAF effect and nerve nutrition, excellent taste sensation.

7 cl, 1 dwg, 5 tbl, 3 ex

Description

FIELD OF TECHNOLOGY

[0001] The invention relates to the technical field of traditional Chinese medicine and, in particular, to the granulated ginkgo biloba ketone ester and a method for producing the same.

PRIOR ART

[0002] As the Chinese population ages, people's awareness of the prevention and treatment of cardiovascular and cerebrovascular diseases is gradually increasing. Ginkgo biloba leaves (ginko biloba) are dry leaves of Ginkgo biloba, sweet, bitter, astringent and soothing, and belonging to the channels of the heart and lungs. According to Chinese Traditional Medicine, ginkgo biloba leaves can "compress lung qi, relieve coughs and asthma, and stop white spots and cloudiness." Ginkgo biloba ketone ester is a ginkgo biloba extract product developed in China. It is an extract of the dried leaves of Ginkgo Biloba L, a plant of the Ginkgo family. Ginkgo biloba ketone ester includes various formulations. Among these preparations, ginkgo biloba granulated ketone ester is light brown-yellow granules, sweet and slightly bitter. Ginkgo biloba ketone ester granular is mainly used to treat dizziness, coronary heart disease and angina pectoris caused by blood congestion type chest obstruction and blood congestion type cerebral arteriosclerosis mild cerebral arteriosclerosis, and to activate and eliminate blood congestion, and its curative effect is significant. Due to the significant curative effect of ginkgo biloba granulated ketone ester, adulteration is a major problem in the production process of related products on the market. For example, flavonoid aglycones such as quercetin are artificially added to significantly increase the detectable content of total flavonol glycosides, which at first glance meets the technical requirements for granular ginkgo biloba ketone ester, but actually affects the internal quality of drugs. Meanwhile, the content of ginkgolic acids in related products on the market is not effectively controlled. Raw materials with a high content of ginkgolic acids are added, which creates a certain safety risk. However, the conventional production methods and quality standards have limitations that make it difficult to ensure the quality stability of the granular ginkgo biloba ketone ester, resulting in a large amount of granular ginkgo biloba ketone ester not being adequate. To ensure the uniformity of different batches of Ginkgo Biloba ketone ester granules, further research and discussion regarding Ginkgo Biloba granulated ketone ester is needed.

BRIEF DESCRIPTION OF THE INVENTION

[0003] The present disclosure provides a granular ginkgo biloba ketone ester and a method for producing the same to solve the problem that conventional granulated ginkgo biloba ketone ester is unstable and poor in quality.

[0004] The first aspect of the present invention relates to granulated ginkgo biloba ketone ester, which is obtained by mixing ginkgo biloba ketone ester raw materials and excipients, granulating and bagging. Granulated ginkgo biloba ketone ester, based on the weight of the load of each package of 1 g, satisfies the following conditions:

[0005] 1) rutin content (C₂₇H_thirtyO₁₆, CAS number 153-18-4) less than or equal to 1.60 mg/pack;

[0006] 2) the content of quercetin (C ₁₅ H ₁₀ O ₇ , CAS number 117-39-5) is less than or equal to 0.160 mg/packet;

[0007] 3) the content of bilobalide (C ₁₅ H ₁₈ O ₈ , CAS number 33570-04-6) is 1.04-2.08 mg/pack;

[0008] 4) the content of ginkgolide J (C ₂₀ H ₂₄ O ₁₀ , CAS No. 107438-79-9) is 0.04-0.20 mg/pack;

[0009] 5) the residual ethanol content is less than or equal to 0.2 mg/pack;

[0010] 6) a biflavonoid content less than or equal to 0.008 mg/packet, said biflavonoids including amentoflavone (C ₃₀ H ₁₈ O ₁₀ , CAS number 1617-53-4), bilobetin (C ₃₁ H ₂₀ O ₁₀ , CAS number: 521 -32-4) and ginkgetin (C ₃₂ H ₂₂ O ₁₀ , CAS number 481-46-9);

[0011] 7) the content of genistin (CAS number 529-59-9) is 0, and the content of ginkgolide M (CAS number 15291-78-8) is 0.

[0012] Preferably, the content of rutin is less than or equal to 1.28 mg/packet.

[0013] Preferably, the content of quercetin is less than or equal to 0.152 mg/packet.

[0014] Preferably, the residual ethanol content is less than or equal to 0.08 mg/packet.

[0015] Preferably, the content of bilobalide is 1.44-1.92 mg/packet.

[0016] Preferably, the content of ginkgolide J is 0.12-0.20 mg/packet.

[0017] Preferably, the content of biflavonoids is less than or equal to 0.004 mg/packet.

[0018] Preferably, the content of total flavonoids in the granulated ginkgo biloba ketone ester is 14.08-26.40 mg/pack when using rutin (C₂₇H_thirtyO₁₆) as a standard sample.

[0019] More preferably, the content of total flavonoids in the granulated ginkgo biloba ketone ester is 17.6-22 mg/bag using rutin as a standard sample.

[0020] More preferably, the content of total flavonoids in the granular ginkgo biloba ketone ester is 19.6-22 mg/bag using rutin as a standard sample. The content of total flavonoids in Ginkgo biloba ketone ester granules is calculated according to Chinese Pharmacopoeia General Regulations 0401 (2015 Edition), Volume IV.

[0021] Preferably, the content of terpene lactones in the granulated ginkgo biloba ketone ester is 2.4-4.8 mg/packet, based on total bilobalide content (C_fifteenH_eighteenO_eight, CAS number 33570-04-6), ginkgolide A (C_twentyH₂₄O₉, CAS number 15291-75-5), ginkgolide B (C_twentyH₂₄O_ten, CAS number 15291-77-7) and ginkgolide C (C_twentyH₂₄O_eleven, CAS number 15291-76-6). The content of terpene lactones is calculated in terms of dry products.

[0022] More preferably, the content of terpene lactones in the granular ginkgo biloba ketone ester is 3.6-4.8 mg/packet, based on the total contents of bilobalide, ginkgolide A, ginkgolide B, and ginkgolide C.

[0023] Preferably, the content of total ginkgolic acid in the granular ginkgo biloba ketone ester is less than or equal to 2 μg/packet.

[0024] More preferably, the content of total ginkgolic acid in the granulated ginkgo biloba ketone ester is less than or equal to 1 μg/packet.

[0025] More preferably, the total ginkgolic acids in the granular ginkgo biloba ketone ester are characterized by total contents of Ginkgolic acid C13:0 (C ₂₀ H ₃₂ O ₃ , CAS number 20261-38-5), Ginkgolic acid C15:1 (C ₂₂ H ₃₄ O ₃ , CAS number 22910-60-7) and Ginkgolic acid C17:1 (C ₂₄ H ₃₈ O ₃ , CAS number 111047-30-4). The content of total ginkgolic acids is calculated in accordance with the General Rules 0512 and 0431 of the Chinese Pharmacopoeia (2015 Edition), Volume IV.

[0026] Preferably, the content of total flavonol glycosides in the granulated ginkgo biloba ketone ester is 9.6-14.0 mg/packet. The content of total flavonol glycosides is calculated in terms of dry products.

[0027] More preferably, the content of total flavonol glycosides in the granulated ginkgo biloba ketone ester is 12.0-14.0 mg/packet.

[0028] More preferably, the total flavonol glycosides are primarily glycosides formed as aglycones by quercetin, kaempferide and isorhamnetin and contain free quercetin, kaempferide and isorhamnetin.

[0029] More preferably, the content of total flavonol glycosides is calculated by the formula (1): content of total flavonol glycosides = (quercetin content + kaempferide content + isorhamnetin content) × 2.51.

[0030] Preferably, the chromatographic peak area ratio of quercetin to kaempferide (C ₁₆ H ₁₂ O ₆ , CAS number 491-54-3) in granular ginkgo biloba ketone ester is 0.8-1.2, and the chromatographic peak area ratio of isorhamnetin ( C ₁₆ H ₁₂ O ₇ , CAS number 207-545-5480-19-3) to quercetin is greater than 0.15.

[0031] The three aglycones (quercetin, kaempferide and isorhamnetin) formed after the hydrolysis of ginkgo flavonoids in granular ginkgo biloba ketone ester have a certain proportional ratio, which to some extent can reflect that the raw materials and manufacturing processes are normal, and can be used for product quality control. The area ratios of the chromatographic peaks of quercetin to kaempferol and isorhamnetin to quercetin were calculated according to conventional methods for the determination of total flavonol glycosides.

[0032] Preferably, the spectrum pattern of the granular ginkgo biloba ketone ester includes four common characteristic peaks as shown in FIG. 1: peak 1 is a characteristic peak of rutin, peak 6 is a characteristic peak of quercetin, peak 7 is a characteristic peak of kaempferide, and peak 8 is a characteristic peak of isorhamnetin.

[0033] Preferably, the similarity between the spectrum pattern of the granulated ginkgo biloba ketone ester and the chromatogram of the test product is greater than or equal to 0.90. The similarity between the spectrum patterns of the test products and Ginkgo Biloba granular ketone ester was calculated and compared according to the "Affinity Scoring System for Chromatographic Patterns in Traditional Chinese Medicine" (Version 2.0).

[0034] The contents of rutin, quercetin, bilobalide, ginkgolide J, total flavonol glycosides, and terpene lactone, as well as the spectrum pattern of the granular ginkgo biloba ketone ester in the above granular ginkgo biloba ketone ester, are determined by high performance liquid chromatography (HPLC) according to the General Rules 0512 in Chinese Pharmacopoeia (2015 Edition) Volume IV. The content of diflavonoids, total ginkgolic acids, ginkgolide M and genistin is determined using high performance liquid chromatography - mass spectrometry (HPLC-MS).

[0035] All of the above percentages (%) are mass percent.

[0036] According to a second aspect of the present invention, there is provided a process for the production of granulated ginkgo biloba ketone ester. The target granulated ginkgo biloba ketone ester is obtained by adding dextrin, sucrose powder, starch, low-substituted hydroxypropyl cellulose and aspartame to the raw material based on ginkgo biloba ketone ester according to the composition ratio, thoroughly mixing, granulating and drying.

[0037] Preferably, the process for producing granulated ginkgo biloba ketone ester includes the following steps:

[0038] A) mixing and kneading ginkgo biloba ketone ester raw materials, dextrin, sucrose powder, starch, low-substituted hydroxypropyl cellulose and aspartame according to the composition ratio and adding ethanol aqueous solution at the same time to obtain a soft material;

[0039] B) obtaining wet granules from the soft material, followed by drying the wet granules;

[0040] C) screening and straightening the dried granules, mixing, tapping and bagging to obtain the desired granular ginkgo biloba ketone ester.

[0041] Preferably in step A), sucrose powder is obtained by grinding sucrose and screening.

[0042] More preferably, the screening number is 70-90 mesh. More preferably, the sieving screen number is 80 mesh.

[0043] Preferably in step A), the starch is corn starch.

[0044] Preferably in step A), the mixing and stirring time is 15-20 minutes.

[0045] Preferably in step A), the aqueous ethanol solution has a weight percentage of 69-71%. The specified 69-71% aqueous ethanol solution is prepared by slowly adding 16.00 ± 0.05 kg of absolute ethanol to 8 kg of purified water with stirring and leave for 5 minutes.

[0046] More preferably, the aqueous ethanol solution has a weight percentage of 70%.

[0047] Preferably in step A), the mixing and mixing is carried out in a mixer. The mixer may be a conventional commercially available mixer. In particular, the mixer is a tray type mixer.

[0048] Preferably in step B), wet granules are produced in a granulator. The granulator may be a conventional commercially available granulator. In particular, the granulator is an oscillating granulator.

[0049] Preferably in step B), wet granules are obtained after screening. More preferably, the screening number is 12-20 mesh. More preferably, the sieving screen number is 16 mesh.

[0050] Preferably in step B), the drying is carried out in a fluid bed dryer. The fluidized bed dryer may be a conventional commercially available fluidized bed dryer.

[0051] More preferably, the air temperature at the entrance to the fluidized bed dryer is 80-105°C.

[0052] More preferably, the fluidized bed dryer inlet air temperature is 90°C.

[0053] Preferably in step B), the drying oscillation time is 80-100 seconds, and the frequency is 5-8 times. More preferably, the drying oscillation time is 90 seconds and the frequency is 6-7 times.

[0054] Preferably in step B), the drying stop temperature of the material is 66-70°C. More preferably, the drying stop temperature of the material is 68°C. The drying stop temperature of a material means that drying is terminated when the predetermined temperature of the material is reached during drying.

[0055] Preferably in step B), the drying time is 15-30 minutes.

[0056] Preferably, in step C), the sieving and straightening screen number is 12-40 mesh.

[0057] Preferably, in step C), the mixing is carried out in a mixer. The mixer may be a conventional commercially available mixer.

[0058] More preferably, the mixer may be a multi-directional mixer.

[0059] Preferably in step C), the agitation speed is 20-25 revolutions per minute (rpm).

[0060] Preferably in step C), the mixing time is 5-6 minutes.

[0061] Preferably, the ratio in the composition of the granulated ginkgo biloba ketone ester, in parts by weight, includes the following:

[0062] 35-45 parts of ginkgo biloba ketone ester raw materials;

[0063] 450-500 parts of dextrin;

[0064] 420-450 parts of sucrose powder;

[0065] 10-30 parts starch;

[0066] 10-30 parts low-substituted hydroxypropyl cellulose; and

[0067] 5-10 parts of aspartame.

[0068] More preferably, the ratio in the composition of the granulated ginkgo biloba ketone ester, in parts by weight, includes the following:

[0069] 40 parts of ginkgo biloba ketone ester raw material;

[0070] 480 parts of dextrin;

[0071] 433 parts of sucrose powder;

[0072] 20 parts starch;

[0073] 20 parts low-substituted hydroxypropyl cellulose; and

[0074] 7 parts of aspartame.

[0075] More preferably, the CAS number of the low substituted hydroxypropyl cellulose is 9004-65-3 or 78214-41-2.

[0076] More preferably, the ginkgo biloba ketone ester feedstock is ginkgo biloba leaf.

[0077] More preferably, ginkgo biloba leaves are dried leaves of ginkgo biloba seedlings. Ginkgo biloba seedling leaves are the leaves of immature ginkgo trees that are planted for the purpose of harvesting ginkgo biloba leaves.

[0078] Still more preferably, the drying conditions are: the drying temperature is 140-160°C, and the drying time is 6-15 minutes.

[0079] Even more preferably, the leaves of the Ginkgo biloba seedlings meet the following requirements:

[0080] a) the content of total flavonol glycosides is greater than or equal to 0.85%;

[0081] b) the content of terpene lactones is greater than or equal to 0.40%;

[0082] c) the total ash content is less than or equal to 10.0%;

[0083] d) the content of acid-insoluble ash is less than or equal to 2.0%;

[0084] e) the content of the extract is greater than or equal to 25.0%;

[0085] f) the chromatographic peak area ratio of quercetin to kaempferide is 0.65-1.2, and the chromatographic peak area ratio of isorhamnetin to quercetin is greater than 0.15;

[0086] g) the content of impurities is less than or equal to 2.0%;

[0087] h) the water content is less than or equal to 12.0%;

[0088] i) a residual sulfur dioxide content of less than or equal to 150 mg/kg.

[0089] The definitions of total flavonol glycosides and terpene lactones in the leaves of ginkgo biloba seedlings are the same as for the granulated ginkgo biloba ketone ester mentioned above. The definition of the area ratio of the chromatographic peaks of flavonoid aglycones in the leaves of Ginkgo biloba seedlings (ie the ratio of the areas of the chromatographic peaks of quercetin, kaempferide and isorhamnetin) is the same as for the aforementioned granulated ginkgo biloba ketone ester.

[0090] The total ash content and the content of acid-insoluble ash in the leaves of Ginkgo biloba seedlings are determined in accordance with the ash determination method described in Chinese Pharmacopoeia General Regulation 2302 (2015 Edition), Volume IV. The content of the extract in the leaves of Ginkgo biloba seedlings is determined in accordance with the method for determining the extract described in the General Rule 2201 of the Chinese Pharmacopoeia (Edition 2015), Volume IV. The content of impurities in the leaves of Ginkgo Biloba seedlings is determined in accordance with the method for determining impurities described in the General Rule 2301 of the Chinese Pharmacopoeia (Edition 2015), Volume IV. The water content of the leaves of Ginkgo Biloba seedlings is determined according to the method for determining water as described in the General Regulations 0832 of the Chinese Pharmacopoeia (2015 Edition), Volume IV. The content of residual sulfur dioxide in the leaves of Ginkgo biloba seedlings is determined in accordance with the method for determining the residual sulfur dioxide described in General Regulations 2331 of the Chinese Pharmacopoeia (Edition 2015), Volume IV.

[0091] Preferably, the process for producing ginkgo biloba ketone ester raw materials includes the following steps:

[0092] 1) adding an aqueous solution of ethanol to the crushed leaves of ginkgo biloba, heating under reflux to extract, and then obtaining a filtrate by filtration;

[0093] 2) concentrating the filtrate and performing aqueous precipitation, centrifuging the supernatant to obtain a centrifuge liquid;

[0094] 3) loading the centrifuge liquid into the macroporous resin column, then washing with water and eluting with a solvent to obtain a first eluent; wherein the first eluent includes a first eluent with a low solvent concentration and a first eluent with a high solvent concentration;

[0095] 4) loading the first eluent with a low solvent concentration into a polyamide column, then washing with water and eluting with ethanol to obtain a second eluent;

[0096] 5) combining and concentrating the second eluent with the first eluent with a high solvent concentration, and then extracting, drying, grinding, sieving and mixing to obtain the target ginkgo biloba ketone ester feedstock.

[0097] More preferably in step 1) ground ginkgo biloba leaves are obtained by grinding ginkgo biloba leaves.

[0098] More preferably, in step 1), obtaining a filtrate includes the following:

[0099] A) adding 60% ethanol aqueous solution to crushed ginkgo biloba leaves, refluxing to extract, and then obtaining a first filtrate by filtration;

[00100] B) adding water to the drug residue remaining after filtration in step A), refluxing to extract, and then obtaining a second filtrate by filtration;

[00101] C) combining and concentrating into a thick paste the second filtrate from step B) and the first filtrate from step A), dissolving, leaving to cool and filtering to obtain the desired filtrate.

[00102] More preferably, in step A), the extraction conditions under reflux are as follows: extraction 2 times; 3 hours for each extraction.

[00103] More preferably in step B) the reflux conditions for extraction are as follows: 1x extraction; 0.5 h for each extraction.

[00104] More preferably in step B) the water is purified water.

[00105] More preferably, in step C) the pooling and concentration is selected from one of the post-combination concentration method or the post-concentration pooling method. The method of concentration after combining is to combine the second filtrate with the first filtrate and then concentrate. The method of combining after concentration is to first concentrate the first filtrate, and then combine with the second filtrate and continue concentration.

[00106] More preferably in step C) the dissolution is a dissolution using hot water. Hot water is purified water heated to 60-100°C.

[00107] Preferably, in step 2), the filtrate is concentrated so that it does not smell of alcohol.

[00108] Preferably in step 2), the relative gravity of the concentrated filtrate is 1.04-1.08.

[00109] Preferably in step 2), the purified water added in the aqueous precipitation is 1.5 to 2.5 times the amount of ginkgo biloba powder. In particular, the fact that the added purified water is 1.5-2.5 times the amount of ginkgo biloba powder means that for every 1 g of ginkgo biloba powder, 1.5-2.5 ml of purified water is added.

[00110] Preferably in step 2), the water precipitation is an intermediate cooling in cooling water at 5-7° C. for 12-24 hours.

[00111] Preferably in step 2), the centrifugation conditions are as follows: the centrifugation speed is 13,000-15,000 rpm and the centrifugation time per 3-5 L of supernatant is 0.75-1.25 minutes.

[00112] More preferably, the centrifugation conditions are as follows: the centrifugation speed is 14,000 rpm and the centrifugation time per 3-5 L of supernatant is 1.00 minutes.

[00113] In step 2), the aforementioned concentration, water settling, and centrifugation can optimize the quality of the ginkgo biloba ketone ester feedstock of the present invention, resulting in better quality of the granulated ginkgo biloba ketone ester.

[00114] Preferably, in step 3), the water wash is first a forward wash with purified water for 1-2 hours and then a backwash for 0.5-1.5 hours. Washing with water can optimize quality conditions such as ginkgolic acids and bottom ash in the ginkgo biloba ketone ester feed of the present invention, resulting in better quality of the granulated ginkgo biloba ketone ester.

[00115] More preferably, the water wash is first a forward wash with purified water for 1.5 hours and then a backwash for 1 hour.

[00116] Preferably, in step 3) the solvent elution is eluting with 18% aqueous ethanol, 30% aqueous ethanol and 50% aqueous ethanol sequentially to obtain 18% ethanol eluent, 30% ethanol eluent and 50% ethanol eluent, respectively.

[00117] Preferably in step 3), the first high solvent concentration eluent is a 50% ethanol eluent.

[00118] Preferably in step 3) the first low solvent concentration eluent is a combined solution of 18% ethanol eluent and 30% ethanol eluent.

[00119] Preferably, in step 4) the water wash is a direct wash using purified water 0.5-2 times the volume of the polyamide column. More preferably, the water wash is a direct wash using 1 times the volume of the polyamide column with purified water. Washing with water further removes impurities, including ginkgolic acid, from the ketone ester of ginkgo biloba.

[00120] Preferably in step 4), the first low solvent concentration eluent is concentrated so that there is no alcohol odor before loading onto the polyamide column.

[00121] Preferably in step 4), the second eluent is an ethanol eluent.

[00122] Preferably in step 5) the pooling and concentration is selected from one of the post-combination concentration method or the post-concentration pooling method. The concentration method after combining is to combine the second eluent with the first eluent with a high solvent concentration and then concentration. The method of combining after concentration is to first concentrate the first eluent with a high concentration of solvent, and then combine it with the second eluent and continue concentration.

[00123] Preferably in step 5), the second eluent and the first eluent with a high solvent concentration are concentrated so that there is no alcohol smell.

[00124] Preferably in step 5), the extraction is carried out using cyclohexane as a solvent. After extraction, the cyclohexane extract is discarded.

[00125] Preferably in step 5), drying is selected from spray drying or vacuum drying.

[00126] More preferably, the concentrated solution must be concentrated to a thick paste prior to vacuum drying.

[00127] Preferably, in step 5), screening is carried out through a sieve of 80-100 mesh. More preferably, screening is carried out through a 90-100 mesh sieve. Most preferably, screening is carried out through a 100 mesh sieve. Screening can ensure that the particle size of the ginkgo biloba ketone ester raw material of the present disclosure is finer, resulting in better quality of the granulated ginkgo biloba ketone ester.

[00128] Preferably in step 5), the mixing conditions are as follows: the mixing equipment is a general mixer; the stirring speed is 10-12 rpm; mixing time 1.5-2.5 hours. Stirring can provide better uniformity of ginkgo biloba ketone ester raw materials according to the present disclosure, resulting in better quality of the granulated ginkgo biloba ketone ester.

[00129] More preferably, the mixing conditions are as follows: the mixing equipment is a general mixer; stirring speed 11 rpm; mixing time 2 hours.

[00130] A third aspect of the present invention relates to the use of a process for producing granular ginkgo biloba ketone ester to produce granular ginkgo biloba ketone ester.

[00131] As mentioned above, provided herein is a granular ginkgo biloba ketone ester and a process for making the same. Thanks to optimized production steps and conditions, high quality Ginkgo Biloba granular ketone ester can be obtained. Compared with the granular ginkgo biloba ketone ester obtained by conventional methods, the granular ginkgo biloba ketone ester of the present invention has a stable quality and a low quality loss rate. The uniformity of different batches of Ginkgo biloba granular ketone ester is good. In particular, the granulated ginkgo biloba ketone ester can be effectively controlled for the presence of ginkgoic acids by preventing the use of raw materials with a high content of ginkgolic acid in the granulated ginkgo biloba ketone ester, in order to effectively improve the safety of the granulated ginkgo biloba ketone ester. At the same time, the granular ginkgo biloba ketone ester obtained by the method of the present invention is excellent in taste, bitterness, stability, high temperature resistance, production characteristics and other characteristics.

BRIEF DESCRIPTION OF GRAPHICS

[00132] FIG. 1 shows the spectrum pattern of the granulated ginkgo biloba ketone ester of the present invention; in spectrum pattern 1: rutin; 6: quercetin; 7: kaempferide; 8: isorhamnetin.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[00133] The present disclosure is further described below with reference to specific embodiments. It should be understood that the embodiments are only intended to describe the present disclosure and not to limit the scope of the present disclosure.

[00134] Embodiments of the present disclosure will be described below. Specialists in the art can easily understand other advantages and effects of the present disclosure in accordance with the content disclosed in the description. The present disclosure may also be implemented or applied using various other illustrative embodiments. Various modifications or changes can also be made to all the details of the description based on different points of view and applications, without going beyond the essence of the present invention.

[00135] It should be noted that process equipment or devices not specifically indicated in the following embodiments are conventional equipment or devices in the art. All pressure values and ranges refer to relative pressures. All reagents used below are conventional reagents in the art. All tools used below are conventional in the art.

[00136] In addition, it should be understood that one or more of the steps of the method mentioned in the present description does not exclude other steps of the method that may be carried out before or after the combined steps, or that other steps of the method can be inserted between these explicitly indicated stages, unless otherwise indicated; it should also be understood that the relationship of the combined connection between one or more of the equipment/devices mentioned in this description does not exclude that there may be other equipment/devices before or after the combined equipment/devices, or that other equipment/devices may be inserted between it. explicitly specified equipment/devices, unless otherwise noted. Moreover, unless otherwise indicated, the numbering of each process step is only a convenient tool for identifying each process step and is not intended to limit the order of each process step or limit the scope of the present disclosure. Changing or adjusting the relative relationship should also be considered as the extent to which the present invention can be implemented without a significant change in technical content.

Embodiment 1

[00137] Dried leaves of ginkgo biloba seedlings are crushed to produce crushed ginkgo biloba leaves. A 60% aqueous ethanol solution was added to crushed ginkgo biloba leaves, extraction was carried out twice by heating under reflux, each time for 3 hours, and the first was obtained by filtration. Then, water is added to the filtered drug residue remaining after filtration, extraction is carried out at one time by reflux with an extraction time of 0.5 hour, and then a second filtrate is obtained by filtration. Combine and concentrate the second filtrate to a thick paste and the first filtrate. Combining and concentrating can be carried out by concentrating after combining or combining after concentration. Dissolved in purified water heated to 60-100°C, left to cool and filtered to obtain a filtrate. The filtrate is concentrated to a relative density of 1.04-1.08 and without the smell of alcohol, purified water is added, which is 1.5-2.5 times the amount of ginkgo biloba leaf powder, cooled in the intermediate layer in cooling water at 5-7 ° C for 12-24 hours and centrifuge the supernatant at 14,000 rpm to obtain centrifuge fluid. The centrifuge liquid is loaded into the macroporous resin column, first, direct washing with purified water is carried out for 1.5 hours, and then backwashing is carried out for 1 hour. Sequential elution is carried out with 18% ethanol aqueous solution, 30% ethanol aqueous solution and 50% ethanol aqueous solution to obtain 18% ethanol eluent, 30% ethanol eluent and 50% ethanol eluent, respectively, which are the first eluents. In the first eluents, 50% ethanol eluent is the first high solvent eluent, and a combined solution of 18% ethanol eluent and 30% ethanol eluent is the first low solvent eluent. Then the first eluent with a low concentration of solvent is concentrated so that there is no smell of alcohol. Load the first eluent with a low solvent concentration into a polyamide column, carry out direct washing with purified water, which is 1 times the volume of the polyamide column, and then elute with ethanol to obtain an ethanol eluate, which is the second eluate. Combine and concentrate the second eluent and the first eluent with a high concentration of solvent until the smell of alcohol disappears. Combining and concentrating can be carried out by concentrating after combining or combining after concentration. After extracting the concentrate with cyclohexane, the cyclohexane extract is discarded. Then spray drying of the concentrated solution or concentration of the concentrated solution into a thick paste and vacuum drying of the thick paste are carried out. The dried products are crushed, passed through a 100 mesh sieve and mixed in a general mixer at a speed of 11 rpm for 2 hours to obtain sample No. 1 of the target raw material based on ginkgo biloba ketone ester.

Test Case 1

[00138] Ordinary ginkgo biloba leaves are ground to produce ground ginkgo biloba leaves. A 60% aqueous ethanol solution was added to crushed ginkgo biloba leaves, extraction was carried out twice by heating under reflux, each time for 3 hours, and a first filtrate was obtained by filtration. Then, water is added to the filtered drug residue remaining after filtration, extraction is carried out at one time by reflux with an extraction time of 0.5 hour, and then a second filtrate is obtained by filtration. Combine and concentrate the second filtrate to a thick paste and the first filtrate. Combining and concentrating can be carried out by concentrating after combining or combining after concentration. Dissolved in purified water heated to 60-100°C, left to cool and filtered to obtain a filtrate. Load the filtrate into a macroporous resin column, elute successively with 18% ethanol aqueous solution, 30% ethanol aqueous solution, and 50% ethanol aqueous solution to obtain 18% ethanol eluent, 30% ethanol eluent, and 50% ethanol eluent, respectively, which are the first eluents. . In the first eluents, 50% ethanol eluent is the first high solvent eluent, and a combined solution of 18% ethanol eluent and 30% ethanol eluent is the first low solvent eluent. Then the first eluent with a low concentration of solvent is concentrated so that there is no smell of alcohol. Load the first eluent with a low concentration of solvent into a polyamide column, elute with ethanol to obtain an ethanol eluate, which is the second eluate. Combine and concentrate the second eluent and the first eluent with a high concentration of solvent until the smell of alcohol disappears. Combining and concentrating can be carried out by concentrating after combining or combining after concentration. After extracting the concentrate with cyclohexane, the cyclohexane extract is discarded. Then spray drying of the concentrated solution or concentration of the concentrated solution into a thick paste and vacuum drying of the thick paste are carried out. The dried products are crushed to obtain a control sample 1* of the target raw material based on the ginkgo biloba ketone ester.

Test Case 2

[00139] Ordinary ginkgo biloba leaves, ginkgo biloba root bark, and Sophorae (Fructus Sophorae) fruits are ground to produce ground ginkgo biloba. A 60% aqueous ethanol solution was added to the crushed ginkgo biloba, extraction was carried out twice by heating under reflux, each time for 3 hours, and a first filtrate was obtained by filtration. Then, water is added to the filtered drug residue remaining after filtration, extraction is carried out at one time by reflux with an extraction time of 0.5 hour, and then a second filtrate is obtained by filtration. Combine and concentrate the second filtrate to a thick paste and the first filtrate. Combining and concentrating can be carried out by concentrating after combining or combining after concentration. Dissolved in purified water heated to 60-100°C, left to cool and filtered to obtain a filtrate. Load the filtrate into a macroporous resin column, elute successively with 18% ethanol aqueous solution, 30% ethanol aqueous solution, and 50% ethanol aqueous solution to obtain 18% ethanol eluent, 30% ethanol eluent, and 50% ethanol eluent, respectively, which are the first eluents. . In the first eluents, 50% ethanol eluent is the first high solvent eluent, and a combined solution of 18% ethanol eluent and 30% ethanol eluent is the first low solvent eluent. Then the first eluent with a low concentration of solvent is concentrated so that there is no smell of alcohol. Load the first eluent with a low concentration of solvent into a polyamide column, elute with ethanol to obtain an ethanol eluate, which is the second eluate. Combine and concentrate the second eluent and the first eluent with a high concentration of solvent until the smell of alcohol disappears. Combining and concentrating can be carried out by concentrating after combining or combining after concentration. After extracting the concentrate with cyclohexane, the cyclohexane extract is discarded. Then spray drying of the concentrated solution or concentration of the concentrated solution into a thick paste and vacuum drying of the thick paste are carried out. The dried products are crushed to obtain a control sample 2* of the target raw material based on the ginkgo biloba ketone ester.

Embodiment 2

[00140] 3.36 kg sample #1 of ginkgo biloba ketone ester raw material, 38.40 kg dextrin, 34.64 kg sucrose powder, 1.60 kg starch, 1.60 kg low-substituted hydroxypropyl cellulose and 0.56 kg aspartame was placed in a tray-type mixer, the mixer was turned on to mix and stir, 70% ethanol was slowly added during mixing, and the mixing and mixing time was 15-20 minutes, to obtain a soft material. Sucrose powder was obtained by grinding sucrose and passing through a 80 mesh sieve. Starch is corn starch. The above feed quantities are in accordance with the quality standard WS _3-227 (Z-028)-2002(Z) and the regulations of the Chinese Pharmacopoeia (2015 Edition) Volume IV, and are within the ratio of Ginkgo biloba granular ketone ester.

[00141] The soft material was placed in an oscillating granulator and passed through a 16 mesh sieve to obtain wet granules. The wet granules were placed in a fluidized bed dryer, the fluidized bed dryer was started, the air temperature at the inlet of the fluidized bed dryer was set to 90°C, dried under conditions where the oscillation time was 90 seconds and the oscillation frequency was 6-7 times, the material temperature was set to 68 °C, drying was stopped when the desired temperature of the material was reached. The total drying time was 15-30 minutes.

[00142] The dried granules were straightened after sifting through a 12-40 mesh shaker screen and then placed in a multi-directional mixer at a rotation speed of 20-25 rpm. After mixing for 5-6 minutes, it was dispensed and packaged in bags to obtain sample #1 of the target granulated ginkgo biloba ketone ester. The load weight of each packet of sample #1 granulated ginkgo biloba ketone ester was 1 g.

Embodiment 3

[00143] 4 kg sample #1 of Ginkgo biloba ketone ester raw material, 48 kg dextrin, 43.30 kg sucrose powder, 2.0 kg starch, 2.0 kg low-substituted hydroxypropyl cellulose and 0.70 kg aspartame were taken into tray-type mixer, the mixing and stirring mixer was started, 70% ethanol was slowly added during mixing, and the mixing and mixing time was 15-20 minutes, to obtain a soft material. Sucrose powder was obtained by grinding sucrose and passing through a 80 mesh sieve. Starch is corn starch. The above feed quantities are in accordance with the quality standard WS _3-227 (Z-028)-2002(Z) and the regulations of the Chinese Pharmacopoeia (2015 Edition) Volume IV, and are within the ratio of Ginkgo biloba granular ketone ester.

[00144] The soft material was placed in an oscillating granulator and passed through a 16 mesh sieve to obtain wet granules. The wet granules were placed in a fluidized bed dryer, the fluidized bed dryer was started, the air temperature at the inlet of the fluidized bed dryer was set to 90°C, dried under conditions where the oscillation time was 90 seconds and the oscillation frequency was 6 to 7 times, the temperature of the material was set to 68°C, drying was stopped when the desired material temperature was reached. The total drying time was 25 minutes.

[00145] The dried granules were straightened after screening through a 12-40 mesh shaker screen and then placed in a multi-directional mixer at a rotation speed of 20-25 rpm. After mixing for 5-6 minutes, it was dispensed and packaged in bags to obtain sample #2 of the target granulated ginkgo biloba ketone ester. The load weight of each packet of sample #2 granulated ginkgo biloba ketone ester was 1 g.

Test Case 3

[00146] 3.36 kg of control sample 1* raw material based on ginkgo biloba ketone ester, 38.40 kg of dextrin, 34.64 kg of sucrose powder, 1.60 kg of starch, 1.60 kg of low-substituted hydroxypropyl cellulose and 0.56 kg of aspartame, mixed well, granulated, dried and bagged according to the conventional method for preparing ginkgo biloba ketone ester granules to obtain a control sample 1* of the target ginkgo biloba ketone ester granules. The loading weight of each packet of control sample ₁ * granular ginkgo biloba ketone ester was 1 g. IV, and are within the ratio in the composition of the granulated ginkgo biloba ketone ester.

[00147] At the same time, 3.36 kg of control sample 2* raw materials based on ginkgo biloba ketone ester, 38.40 kg of dextrin, 34.64 kg of sucrose powder, 1.60 kg of starch, 1.60 kg of low-substituted hydroxypropyl cellulose and 0.56 kg of aspartame, mixed well, granulated, dried and bagged according to the conventional method for preparing granulated ginkgo biloba ketone ester to obtain a control sample 2* of the target granulated ginkgo biloba ketone ester. The loading weight of each packet of control sample ₂ * granular ginkgo biloba ketone ester was 1 g. IV, and are within the ratio in the composition of the granulated ginkgo biloba ketone ester.

Embodiment 4

[00148] 10 packets containing Ginkgo Biloba Granulated Ketone Ester Sample #1 (obtained in Embodiment 2), Control Samples 1* and 2* Ginkgo Biloba Ketone Ester Granular (obtained in Control Example 3) were taken, respectively, mixed well, finely ground and selected 2 g of the powder. The powder was accurately weighed, the powder was placed in a stoppered conical flask, 10 ml of a mixed solution of 70% methanol-70% ethanol (1:1) was accurately added, and extracted by shaking (500 times per minute) for 30 minutes. The solution was taken for centrifugation to obtain the supernatant (or the solution was filtered to obtain subsequent filtrates) to obtain solutions of the test product A1, A2 and A3.

[00149] At the same time, appropriate amounts of rutin reference substance and quercetin reference substance were taken, and said reference substances were accurately weighed. Methanol was added to the reference substances to obtain a mixed solution containing 250 μg of rutin and 25 μg of quercetin per 1 ml to obtain a reference solution.

[00150] 10 μl were accurately pipetted from the test product solutions A1, A2, A3 and reference solution, respectively, which were then analyzed by high performance liquid chromatography (HPLC) according to the General Rule 0512 of the Chinese Pharmacopoeia (2015 Edition), Volume IV. High performance liquid chromatography conditions are: silica gel chemically bonded with octadecylsilane serves as a filler; acetonitrile served as mobile phase A and 0.4% phosphoric acid served as mobile phase B. Gradient elution was performed as described in Table 1 below; the detection wavelength was 400 nm. The number of theoretical plates calculated from the rutin peak was at least 8000.

Table 1

Time (minutes) Mobile phase A (%) Mobile phase B (%) 0~35 fifteen 85 35~80 twenty 80

[00151] The result of the determination shows that in sample #1 granular ginkgo biloba ketone ester, the content of rutin is 1.07 mg/pack, and this value is less than or equal to 1.28 mg/pack; the content of quercetin is 0.132 mg/package, and this value is less than or equal to 0.152 mg/package. In the control sample 1* granulated Ginkgo biloba ketone ester, the content of rutin is 1.42 mg/pack and this value is less than or equal to 1.60 mg/pack; the content of quercetin is 0.157 mg/package, and this value is less than or equal to 0.160 mg/packet. In the control sample 2* granulated Ginkgo biloba ketone ester, the content of rutin is 1.82 mg/bag, and this value is greater than 1.60 mg/bag; the content of quercetin is 0.183mg/package, and this value is more than 0.160mg/package. From the above content values, it can be seen that the content of rutin and quercetin in sample #1 Ginkgo biloba ketone granular ester and in control sample 1* Ginkgo biloba granular ketone ester is within the specified range of values, indicating the absence of foreign contaminants, normal the flow of the production process and the good quality of the product. The contents of rutin and quercetin in sample #1 granular ginkgo biloba ketone ester are lower than in control sample 1* granular ginkgo biloba ketone ester, indicating that the product quality of sample #1 granulated ginkgo biloba ketone ester is better than the control sample 1* granulated ginkgo biloba ketone ester. The levels of rutin and quercetin in the control sample 2* granular ginkgo biloba ketone ester exceed the indicated range of values, indicating the possible presence of foreign contaminants and poor quality of the product.

Embodiment 5

[00152] 10 sachets containing Ginkgo Biloba Granular Ketone Ester Sample #1 (obtained in Embodiment 2), Controls 1* and 2* Ginkgo Biloba Ketone Ester Granular (obtained in Control Example 3) were taken, respectively, mixed well, finely crushed, 1 g of powder was taken and dried by spraying. The samples were dried in a vacuum oven at a temperature less than or equal to 80°C for 2 hours; or the liquid extract was concentrated to a relative density of 1.2 followed by drying at an inlet air temperature of 160-165°C and an outlet air temperature of 95-100°C.

[00153] The determination result shows that sample #1 of ginkgo biloba granulated ketone ester has an ethanol residue content of 0.06 mg/packet and is less than or equal to 0.08 mg/packet. In the control sample 1* Ginkgo biloba ketone granular ester, the content of ethanol residue is 0.15 mg/pack, and this value is less than or equal to 0.2 mg/pack. In the control sample 2* Ginkgo biloba ketone granular ester, the content of ethanol residue is 0.18 mg/pack, and this value is less than or equal to 0.2 mg/pack. It can be seen from the above contents that the ethanol residues in Sample #1 of Ginkgo Biloba ketone granular ester and Control Samples 1* and 2* of Ginkgo Biloba granular ketone ester all met the control requirements. However, Ginkgo Biloba Granulated Ketone Ester Sample #1 has the least residual ethanol and the best quality. Control samples 1* and 2* Ginkgo biloba ketone ester granules have almost the same amount of residual ethanol, but the amount of residual ethanol is higher than sample #1 Ginkgo biloba ketone ester granules, therefore, is of poorer quality.

Embodiment 6

[00154] 10 sachets containing Ginkgo Biloba Granulated Ketone Ester Sample #1 (obtained in Embodiment 2), Controls 1* and 2* Ginkgo Biloba Ketone Ester Granular (obtained in Control Example 3) were taken, respectively, mixed well, finely ground and selected 3 g of powder. Accurately weigh the powder, put the powder into a 50 ml centrifuge tube with a stopper, add 15 ml of 30% ethanol, shake well, add 20 ml of diethyl ether, shake well, leave for a while, carefully open the lid, press the lid after air is released, shake (3000 times per minute) for 1 minute, centrifuged (4000 rpm) for 10 minutes, the supernatant was separated. Added diethyl ether to the residue, repeated the above operations 3 times, each time adding 15 ml of diethyl ether. The four diethyl ether solutions were combined, concentrated under reduced pressure to almost dryness (not evaporating to dryness), the proper amount of methanol was added and dissolved by sonication, and completely transferred to a 5 ml volumetric flask. Centrifuge to obtain the supernatant (or pass through a 0.45 μm filter membrane to obtain subsequent filtrates) to obtain solutions of test product B1, B2 and B3.

[00155] At the same time, the respective amounts of bilobalide reference substance, ginkgolide A reference substance, ginkgolide B reference substance, ginkgolide C reference substance, and ginkgolide J reference substance were taken and accurately weighed. Methanol was added to the reference substances to prepare mixed solutions, each containing 1.0, 0.3, 0.7 and 0.4 mg of each reference substance per 1 ml, which is the reference solution.

[00156] 5 µl and 10 µl of reference solution and 10 µl of test product solutions B1, B2, and B3, respectively, were accurately pipetted and then analyzed by high performance liquid chromatography (HPLC) according to Chinese Pharmacopoeia General Rule 0512 (2015 edition), volume IV. The contents of bilobalide, ginkgolide A, ginkgolide B, ginkgolide C and ginkgolide J were calculated using an external standard two-point logarithmic equation. High performance liquid chromatography conditions are: silica gel chemically bonded with octadecylsilane serves as a filler; methanol serves as mobile phase A and water serves as mobile phase B. Gradient elution was performed as described in Table 2 below; detection was performed with an evaporative light scattering detector. The number of theoretical plates calculated from the bilobalide peak was at least 10,000.

table 2

Time (minutes) Mobile phase A (%) Mobile phase B (%) 0~15 thirty 70 15~30 40 60

[00157] The determination results show that the terpene lactone contents of Ginkgo Biloba Granulated Ketone Ester Sample #1 and Control Sample 1* Ginkgo Biloba Ketone Ester Granular Essence are calculated on a dry basis and are calculated from the total contents of Bilobalide, Ginkgolide A, Ginkgolide B and Ginkgolide C. The contents of terpene lactones in sample #1 Ginkgo biloba ketone granular ester and control sample 1* Ginkgo biloba granulated ketone ester are 4.4 mg/bag and 2.9 mg/bag, respectively, which are within the indicated range is 2.4-4.8 mg/packet. The terpene lactone content of Control 2* Granulated Ginkgo Biloba Ketone Ester is calculated on a dry basis and is calculated from the total contents of Bilobalide, Ginkgolide A, Ginkgolide B and Ginkgolide C. Terpene lactone content of Control 2* Granulated Ginkgo Ketone Ester biloba is 1.8 mg/pack, which is below the stated range of 2.4-4.8 mg/pack. Terpene lactones, as unique components of ginkgo biloba, are the main effective components of ginkgo biloba granular ketone ester for the activation and elimination of blood stasis, as well as in the treatment of angina pectoris, coronary heart disease and cerebral arteriosclerosis. Therefore, terpene lactones must be within the specified data range. It can be seen that the control sample 2* Ginkgo biloba granulated ketone ester has little curative effect. Sample #1 Ginkgo biloba ketone granular ester and control sample 1* Ginkgo biloba granular ketone ester have good therapeutic effects. However, the content of terpene lactones in Sample #1 Ginkgo Biloba Granulated Ketone Ester is higher, which is within the stated range of 3.6-4.8 mg/packet. Compared with the control sample 1* Ginkgo biloba ketone granular ester, sample #1 Ginkgo biloba ketone granular ester has a better therapeutic effect.

[00158] At the same time, the detection results show that the content of bilobalide in sample #1 granular ginkgo biloba ketone ester and in control sample 1* granular ginkgo biloba ketone ester is 1.78 mg/bag and 1.17 mg/ package, respectively, which is within the specified range of 1.04-2.08 mg/package. The content of bilobalide in the control sample 2* granulated ginkgo biloba ketone ester is 0.96 mg/pack, which is below the specified range of 1.04-2.08 mg/pack. Bilobalide gives Ginkgo biloba granular ketone ester an anti-PAF (platelet activating factor) effect and also nourishes the nerves. Therefore, the control sample 2* Ginkgo biloba granulated ketone ester has little curative effect. Sample #1 Ginkgo biloba ketone granular ester and control sample 1* Ginkgo biloba granular ketone ester have good therapeutic effects. However, the content of bilobalide in sample #1 of Ginkgo biloba granulated ketone ester is higher, which is within the specified range of 1.44-1.92 mg/packet. Compared with the control sample 1* Ginkgo biloba ketone granular ester, sample #1 Ginkgo biloba ketone granular ester has a better therapeutic effect.

[00159] At the same time, the determination results show that the contents of ginkgolide J in sample #1 granular ginkgo biloba ketone ester and in control sample 1* granular ginkgo biloba ketone ester are 0.15 mg/bag and 0.10 mg /package, respectively, which is within the specified range of 0.04-0.20 mg/package. The content of ginkgolide J in the control sample 2* granular ginkgo biloba ketone ester is 0.02 mg/pack, which is below the specified range of 0.04-0.20 mg/pack. Ginkgolide J also imparts an anti-PAF effect to Ginkgo Biloba granular ketone ester. Therefore, the control sample 2* Ginkgo biloba granulated ketone ester has little curative effect. Sample #1 Ginkgo biloba ketone granular ester and control sample 1* Ginkgo biloba granular ketone ester have good therapeutic effects. However, the content of ginkgolide J in sample #1 of Ginkgo biloba granular ketone ester is higher, which is within the specified range of 0.12-0.20 mg/packet. Compared with the control sample 1* Ginkgo biloba ketone granular ester, sample #1 Ginkgo biloba ketone granular ester has a better therapeutic effect.

Embodiment 7

[00160] 10 sachets containing Ginkgo Biloba Granular Ketone Ester Sample #1 (prepared in Variant 2), Control Samples 1* and 2* Ginkgo Biloba Ketone Ester Granular (prepared in Control Example 3) were taken, respectively, good mixed, finely crushed and selected 0.1 g of powder. The powder was accurately weighed, sonicated with 5 ml ethanol for 10 minutes, centrifuged at 3000 rpm for 10 minutes, and then the supernatant was collected. The above ultrasonic extraction was repeated 3 times. The supernatants were pooled and made up to volume with a 25 ml volumetric flask to give test product solutions C1, C2 and C3.

[00161] At the same time, the respective amounts of amentoflavone reference substance, bilobetin reference substance, and ginkgetin reference substance were taken and accurately weighed. Ethanol was added to the reference substances to obtain a mixed solution with a certain concentration of amentoflavone, bilobetin and ginkgetin, which is the reference solution.

[00162] Accurately pipet 10 µl from test product solutions C1, C2, C3 and reference solution, respectively, then analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS) according to Chinese Pharmacopoeia General Rules 0512 and 0431 (2015 Edition). ), volume IV. HPLC-MS conditions are: silica gel chemically bound with octadecylsilane, which serves as a filler; a solution of methanol and 1% glacial acetic acid (90:10), which serves as the mobile phase, was eluted isocratically, completely purified with a solution of methanol and 1% glacial acetic acid (99:1), which is at least 10 times the volume of the column when all determined components reached the peak; a triple quadrupole mass spectrometer was used to monitor multiple reactions (MRM) in electrospray ionization (ESI) mode with negative ions.

[00163] The results of the assay show that the biflavonoid content of Sample #1 Ginkgo Biloba Granulated Ketone Ester and Control Sample 1* Granulated Ginkgo Biloba Ketone Ester is 0.002 mg/packet and 0.007 mg/packet, respectively, i.e. all total amentoflavone contents , bilobetin and ginkgetin in Sample #1 Ginkgo Biloba Granulated Ketone Ester and Control Sample 1* Ginkgo Biloba Granular Ketone Ester less than or equal to 0.008 mg/packet. The content of biflavonoids in the control sample 2* granular ginkgo biloba ketone ester is 0.013 mg/pack, i.e. the total contents of amentoflavone, bilobetin and ginkgetin in the control sample 2* granulated ginkgo biloba ketone ester exceed 0.008 mg/pack. The content of biflavonoids must be controlled due to their allergenicity. The content of biflavones in the control sample 2* Ginkgo biloba granulated ketone ester exceeds the limit, which makes patients more prone to allergies. The content of biflavonoids in sample #1 granular ginkgo biloba ketone ester and control sample 1* granulated ginkgo biloba ketone ester is within the established limits. However, Ginkgo Biloba Granulated Ketone Ester Sample #1 has a lower biflavonoid content, which can be as low as 0.004 mg/pouch or less, and is less likely to cause sensitization.

Embodiment 8

[00164] 10 packets containing Ginkgo biloba ketone granular ester sample #1 (prepared in Variant 2), Controls 1* and 2* Ginkgo biloba ketone granular ester (prepared in Control 3) were taken, respectively, good mixed, finely crushed and selected 0.1 g of powder. The powder was accurately weighed, extracted with sonication with 5 ml of 50% ethanol for 10 minutes, centrifuged at 3000 rpm for 10 minutes, and then the supernatant was collected. The above ultrasonic extraction was repeated 3 times. The supernatants were pooled and made up to volume with a 25 ml volumetric flask to give test product solutions D1, D2 and D3.

[00165] At the same time, appropriate amounts of genistin reference substance and ginkgolide M reference substance, respectively, were taken, and said reference substances were accurately weighed. Methanol was added to the reference substances to prepare a mixed solution with a certain concentration of genistin and ginkgolide M, which is the control solution.

[00166] 10 μl were accurately pipetted from the test product solutions D1, D2, D3 and the reference solution, respectively, then analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS) according to Chinese Pharmacopoeia General Rules 0512 and 0431 (2015 Edition). ), volume IV. HPLC-MS conditions are: silica gel chemically bonded with octadecylsilane serves as filler; a solution of methanol and 1% glacial acetic acid (90:10) serves as the mobile phase, eluted isocratically, completely purified with a solution of methanol and 1% glacial acetic acid (99:1) which is at least 10 times the volume of the column when all determined components reached a peak; a triple quadrupole mass spectrometer was used to monitor multiple reactions (MRM) in electrospray ionization (ESI) mode with negative ions.

[00167] It can be seen from the detection results that neither Ginkgo Biloba Granulated Ketone Ester Sample #1 nor Control Ginkgo Biloba Ketone Ester Granulated Ester 1* contains genistin and ginkgolide M. However, Control Sample 2* Ginkgo Biloba Ketone Ester Granular contains genistin and ginkgolide M. Genistin is a component present in Sophorae fruit, ginkgolide M is a component present in ginkgo biloba root bark, none of the above components are present in ginkgo biloba leaves. Thus, the detection of the above components in the control 2* granulated ginkgo biloba ketone ester means that undesirable foreign substances, such as Sophorae fruits and ginkgo biloba root bark, are inappropriately present in the control 2* granulated ginkgo biloba ketone ester. Sample #1 Ginkgo Biloba Ketone Ester Granular and Control Sample 1* Ginkgo Biloba Ketone Ester Granular are free of unwanted foreign matter, indicating good product quality.

Embodiment 9

[00168] We took 20 mg of the rutin reference substance, weighed it precisely, placed the rutin reference substance in a 100 ml volumetric flask, added 70 ml of 70% ethanol, slightly heated in a water bath to dissolve, cooled, diluted to the mark with 70% ethanol and well shaken to obtain a reference solution (containing 0.2 mg anhydrous rutin per ml).

[00169] Then, accurately measured 0.2 ml, 0.4 ml, 0.6 ml, 0.8 ml, 1.0 ml and 1.2 ml of the standard solution, put the standard solution into 10 ml volumetric flasks, respectively , water was added to 3 ml, 2 ml of sodium acetate buffer (pH 4.5) and 2 ml of 0.1 mol/l aluminum chloride solution were added, shaken well, 70% ethanol was added to the mark and shaken well. The corresponding solution served as blank solution. The tests were carried out by ultraviolet-visible spectrophotometry (Chinese Pharmacopoeia General Regulations 0401 (2015 Edition), Volume IV). Optical density was measured at a wavelength of 270 nm, and a standard curve was built with optical density on the ordinate axis and concentration on the abscissa axis.

[00170] 10 packets containing Ginkgo Biloba Granular Ketone Ester Sample #1 (obtained in Embodiment 2), Control Samples 1* and 2* Ginkgo Biloba Ketone Ester Granular (obtained in Control Example 3) were taken, respectively, mixed well, finely crushed, and 1 g of powder was taken. The powder was accurately weighed, the powder was placed in Soxhlet extractors, 40 ml of 70% ethanol was added, heated on a water bath and heated under reflux for 4 hours, cooled, the extract was transferred into 50 ml volumetric flasks, diluted to the mark with 70% ethanol and well shaken. Accurately measure 0.5 ml into a 10 ml volumetric flask, add water to 3 ml, add 2 ml of sodium acetate buffer (pH 4.5) and 2 ml of 0.1 mol/l aluminum chloride solution, shake well, add 70 % ethanol to the mark and shake well. The tests were carried out by ultraviolet-visible spectrophotometry (Chinese Pharmacopoeia General Regulations 0401 (2015 Edition), Volume IV). Optical density was measured at a wavelength of 270 nm. The equivalent mass of rutin in the test product solution was read from the standard curve, which is calculated to determine the content of total flavonoids.

[00171] The results of the determination show that when using rutin (C₂₇H_thirtyO₁₆) as a standard sample, the content of total flavonoids in Sample #1 of Ginkgo Biloba Granular Ketone Ester and Control Sample 1* of Ginkgo Biloba Granular Ketone Ester is 21.40 mg/bag and 15.72 mg/bag, respectively, which is within the indicated range is 14.08-26.40 mg/packet. The content of total flavonoids in the control sample 2* granulated ginkgo biloba ketone ester is 12.43 mg/pack when using rutin (C₂₇H_thirtyO₁₆) as a standard sample, which is below the specified range of 14.08-26.40 mg/bag. Total flavonoids are active substances which, in addition to the total flavonol glycosides, contain free flavonoids. The certain amount of total flavonoids in Ginkgo Biloba Ketone Ester Granular provides a good therapeutic effect of Ginkgo Biloba Ketone Ester Granular. Therefore, the control sample 2* Ginkgo biloba granular ketone ester has little curative effect. Sample #1 Ginkgo biloba granular ketone ester and control sample 1* Ginkgo biloba granular ketone ester have good therapeutic effects. However, the content of total flavonoids in sample #1 granulated ginkgo biloba ketone ester is higher, which is within the specified range of 19.6-22 mg/packet. Compared with the control sample 1* Ginkgo biloba ketone ester granules, sample #1 Ginkgo biloba ketone ester granules has a better therapeutic effect.

Embodiment 10

[00172] 10 packets containing Ginkgo Biloba Granulated Ketone Ester Sample #1 (obtained in Embodiment 2), Controls 1* and 2* Ginkgo Biloba Ketone Ester Granular (obtained in Control Example 3) were taken, respectively, mixed well, finely ground and selected 2 g of the powder. The powder was accurately weighed, the powder was placed in a stoppered conical flask, 10 ml of methanol was accurately added, weighed, sonicated (power: 180 W; frequency: 42 kHz) for 20 minutes, cooled, reweighed, methanol was used to compensate for the weight loss. Filtered and collected the subsequent filtrate to obtain solutions of the test product D1, D2 and D3.

[00173] At the same time, appropriate amounts of C13:0 Ginkgolic Acid Reference Substance, C15:1 Ginkgolic Acid Reference Substance, and C17:1 Ginkgolic Acid Reference Substance, respectively, were taken, and the reference substances were accurately weighed. Methanol was added to the reference substances to prepare a series of mixed solutions containing 0 ng/ml, 10 ng/ml, 20 ng/ml, 50 ng/ml, 100 ng/ml and 200 ng/ml, respectively, of each reference substance to obtain a reference solution .

[00174] Accurately pipet 1 µl from test product solutions D1, D2, D3 and reference solutions, respectively, then analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS) according to Chinese Pharmacopoeia General Rules 0512 and 0431 (2015 Edition). ), volume IV. HPLC-MS conditions are: silica gel chemically bonded with octadecylsilane serves as filler; a solution of methanol and 1% glacial acetic acid (90:10) serves as the mobile phase, eluted isocratically, completely purified with a solution of methanol and 1% glacial acetic acid (99:1) which is at least 10 times the volume of the column when all determined components reached a peak; a triple quadrupole mass spectrometer was used to monitor multiple reactions (MRM) in electrospray ionization (ESI) mode with negative ions. The ion pairs that were monitored are shown in Table 3 below. If a chromatographic peak with the same retention time as that of the reference substance is found in the test product solution, and the selected relative ion content and the relative ion content of the corresponding reference solution concentration meet the requirements of Table 4 below, the component can be determined to be present.

Table 3

Name precursor ion Quantitative ion pair Quality ion pair Ginkgolic acid C13:0 319.2 319.2 → 275.2 319.2 → 106.1 Ginkgolic acid C15:1 345.2 345.2 → 301.2 345.2 → 119.0 Ginkgolic acid C17:1 373.3 373.3 → 329.3 373.3 → 106.0

Table 4

Relative content of ions/% more than 50 20~50 10~20 less than 10 Permissible relative deviation /% ±20 ±25 ±30 ±50

[00175] The results of the determination show that the content of total ginkgolic acids in sample #1 granular ginkgo biloba ketone ester and control sample 1* granulated ginkgo biloba ketone ester is 0.7 μg/bag and 1.5 μg/bag, respectively, which is less than or equal to 2 mcg/pack. The content of total ginkgolic acids in the control sample 2* granulated ginkgo biloba ketone ester is 3.1 µg/pack, which is more than 2 µg/pack. Ginkgolic acid has a sensitizing effect. Long-term use of the drug ginkgo can cause the accumulation of ginkgolic acid and harm the human body. Therefore, it is necessary to control the content of ginkgoic acids in the granular ginkgo biloba ketone ester so as to keep the content of ginkgolic acids in the granular ginkgo biloba ketone ester low, prevent the use of high ginkgolic acid raw materials for the granulated ginkgo biloba ketone ester, and improve the safety of the granulated ginkgo biloba ketone ester, which has never been controlled in traditional granular ginkgo biloba ketone ester. The content of total ginkgolic acids in the control sample of 2* Ginkgo biloba ketone ester granules exceeds the specified limit, which may harm the human body. The contents of total ginkgolic acids in sample #1 granulated ginkgo biloba ketone ester and control sample 1* granular ginkgo biloba ketone ester are within the established limits. However, Ginkgo Biloba Granulated Ketone Ester Sample #1 has a lower total ginkgolic acid content, which can be less than or equal to 1 µg/pouch, and is less likely to cause sensitization and is therefore safer.

Embodiment 11

[00176] 10 packets containing Ginkgo Biloba Granular Ketone Ester Sample #1 (obtained in Embodiment 2), Control Samples 1* and 2* Ginkgo Biloba Ketone Ester Granular (obtained in Control Example 3) were taken, respectively, mixed well, finely crushed, and 1 g of powder was taken. Accurately weighed the powder, accurately added 50 ml of a mixed solution of methanol and 25% hydrochloric acid solution (4:1), weighed, placed in a water bath of 85-90°C and heated under reflux for 30 minutes, removed, quickly cooled to room temperature. temperature, weighed again using the above mixed solution to make up for the weight loss, and shaken well. Filtered and collected the subsequent filtrate to obtain solutions of the test product E1, E2 and E3 (must be tested within 3 hours).

[00177] At the same time, appropriate amounts of quercetin reference substance, kaempferide reference substance, and isorhamnetin reference substance were taken, and the reference substances were accurately weighed. Methanol was added to the reference substances to obtain mixed solutions containing respectively 30 μg, 30 μg, 20 μg of each reference substance per 1 ml to obtain a reference solution.

[00178] 10 µl were accurately pipetted from the test product and reference solutions, respectively, which were then analyzed by high performance liquid chromatography (HPLC) according to Chinese Pharmacopoeia General Rule 0512 (2015 Edition), Volume IV. High performance liquid chromatography conditions are: silica gel chemically bonded with octadecylsilane as a filler; methanol with 0.4% phosphoric acid (49:51) served as the mobile phase; eluted isocratically; the detection wavelength is 368 nm. The number of theoretical plates, based on the peak of quercetin, was not less than 4000. The degree of separation of the kaempferide peak and the isorhamnetin peak should be greater than 1.5.

[00179] The content of total flavonol glycosides in the ketone ester of ginkgo biloba is calculated by the formula (1): content of total flavonol glycosides = (quercetin content + kaempferide content + isorhamnetin content) × 2.51.

[00180] The determination results show that the total flavone glycosides in Sample #1 Ginkgo Biloba Granulated Ketone Ester and Control Sample 1* Ginkgo Biloba Granular Ketone Ester are 13.2 mg/pouch and 10.3 mg/pouch (based on dry products), respectively, which is within the specified range of 9.6-14.0 mg/packet. The content of total flavonol glycosides in the control sample 2* granulated ginkgo biloba ketone ester is 7.6 mg/pack (based on dry products), which is below the specified range of 9.6-14.0 mg/pack. General flavonol glycosides are active substances containing free flavonoids. A certain amount of total flavone glycosides in Ginkgo biloba ketone ester granules provides a good therapeutic effect of ginkgo biloba ketone ester granules. Therefore, the control sample 2* Ginkgo biloba granulated ketone ester has little curative effect. Sample #1 Ginkgo biloba ketone granular ester and control sample 1* Ginkgo biloba granular ketone ester both have good therapeutic effects. However, the content of total flavone glycosides in Sample #1 of Ginkgo biloba granulated ketone ester is higher, which is within the specified range of 12.0-14.0 mg/packet. Compared with the control sample 1* Ginkgo biloba ketone granular ester, sample #1 Ginkgo biloba ketone granular ester has a better therapeutic effect.

[00181] 10 μl of test product solutions E1, E2, E3 and reference solution, respectively, were accurately pipetted, which were then analyzed using the aforementioned high performance liquid chromatography (HPLC). The determination results show that in sample #1 Ginkgo biloba ketone granular ester and control sample 1* Ginkgo biloba granular ketone ester, the chromatographic peak area ratios of quercetin to kaempferide are 0.90 and 1.01, respectively, which are within the specified range of 0 .8-1.2; the area ratios of the chromatographic peaks of isorhamnetin to quercetin are 0.20 and 0.21, respectively, where both values are greater than 0.15. In the control sample 2* granulated ginkgo biloba ketone ester, the ratio of the areas of chromatographic peaks of quercetin to kaempferide is 0.59, which is below the specified range of 0.8-1.2; the area ratio of the chromatographic peaks of isorhamnetin to quercetin is 0.12, which is below 0.15. From the above ratio, it can be seen that Sample No. 1 Ginkgo Biloba Ketone Ester Granular and Control Sample 1* Ginkgo Biloba Ketone Ester Granular have no unwanted foreign matter, indicating a normal production process and good product quality. The quality of the control sample 2* granulated ginkgo biloba ketone ester is poor due to the addition of quercetin and undesirable foreign substances.

Embodiment 12

[00182] 10 sachets containing Ginkgo biloba ketone ester granular sample #1 (obtained in Embodiment 2), control samples 1* and 2* ginkgo biloba granulated ketone ester (obtained in Control Example 3) were taken, respectively, mixed well, finely crushed, and 1 g of powder was taken. Powder was accurately weighed, 10 ml of 75% methanol was added, sonicated (power: 300 W, frequency: 50 kHz) for 10 minutes, centrifuged for 5 minutes (4000 rpm), supernatant was collected to obtain solutions of the test product F1, F2 and F3.

[00183] At the same time, appropriate amounts of rutin reference substance, quercetin reference substance, kaempferide reference substance, and isorhamnetin reference substance were taken, and the reference substances were accurately weighed. 75% methanol was added to the reference substances to obtain a solution containing 30 µg of each reference substance per 1 ml, and a reference solution was obtained.

[00184] 10 μl were accurately pipetted from the test product and reference solutions, respectively, which were then analyzed by high performance liquid chromatography (HPLC) according to Chinese Pharmacopoeia General Rule 0512 (2015 Edition), Volume IV. The high performance liquid chromatography conditions are: octadecylsilane chemically bonded silica gel serves as a filler (column length: 15 cm; inner diameter: 4.6 mm; particle size: 5 μm); acetonitrile served as mobile phase A and 0.1% formic acid served as mobile phase B. Gradient elution was performed according to the specification in Table 5 below; the detection wavelength is 360 nm; column temperature 30°C; flow rate 1.0 ml/min. The number of theoretical plates calculated from the rutin peak is at least 10,000.

Table 5

Time (minutes) Mobile phase A (%) Mobile phase B (%) 0-40 15-30 85-70 40-45 30-40 70-60 45-50 40 60 50-60 40-60 60-40 60-70 60 40

[00185] The similarity between the spectrum patterns of test products and granular ginkgo biloba ketone ester was calculated and compared according to the "Affinity Scoring System for Chromatographic Patterns in Traditional Chinese Medicine" (Version 2.0). The spectrum pattern of the granular ginkgo biloba ketone ester includes four common characteristic peaks, as shown in FIG. 1: peak 1 is a characteristic peak of rutin, peak 6 is a characteristic peak of quercetin, peak 7 is a characteristic peak of kaempferide, and peak 8 is a characteristic peak of isorhamnetin. The determination results show that the similarity between the spectrum patterns of sample #1 granular ginkgo biloba ketone ester and control sample 1* granulated ginkgo biloba ketone ester and the chromatogram of the test product is 0.97 and 0.91, respectively, where both values are greater than or equal to 0.90. The similarity between the spectrum pattern of the control sample 2* granulated ginkgo biloba ketone ester and the chromatogram of the test product is 0.31, which is below 0.90. It can be seen from the above similarities that sample #1 Ginkgo biloba ketone ester granules and control sample 1* Ginkgo biloba ketone ester granules are free of unwanted foreign matter, indicating good product quality. The quality of the control sample 2* granular ginkgo biloba ketone ester is poor due to the presence of undesirable foreign substances.

Embodiment 13

[00186] The leaves of Ginkgo biloba seedlings, which serve as the medicinal raw material in Embodiment 1, are selected and dried at 140-160°C for 6-15 minutes to serve as the raw material of Sample 1; at the same time, ordinary ginkgo biloba leaves, which served as the medicinal raw material in Control Example 1, were selected as the raw material of Sample 2.

[00187] The contents of total flavonol glycosides and the ratio of the areas of chromatographic peaks of flavonoid aglycones in the Sample 1 feed and Sample 2 feed are determined in accordance with the method described in Embodiment 9. The terpene lactone contents in the Sample 1 feed and Sample 2 feed are determined by the method described in Embodiment implementation 4. The total ash content and acid-insoluble ash in sample 1 raw material and sample 2 raw material are determined in accordance with the ash determination method described in Chinese Pharmacopoeia General Regulation 2302 (2015 Edition), Volume IV. The extract contents of sample 1 raw material and sample 2 raw material are determined according to the extract determination method described in Chinese Pharmacopoeia General Regulations 2201 (2015 Edition), Volume IV. The impurity content of Sample 1 Raw Material and Sample 2 Raw Material is determined according to the impurity determination method described in Chinese Pharmacopoeia General Regulation 2301 (2015 Edition) Volume IV. The water contents of sample 1 feedstock and sample 2 feedstock are determined according to the water determination method described in Chinese Pharmacopoeia General Regulation 0832 (2015 Edition), Volume IV. The content of residual sulfur dioxide in raw material sample 1 and raw material sample 2 is determined in accordance with the method for determining the residual sulfur dioxide described in General rules 2331 of the Chinese Pharmacopoeia (2015 edition), volume IV.

[00188] The results of the determination show that the content of total flavonol glycosides in the raw material sample 1 and raw material sample 2 is 1.34% and 0.54%, respectively. The content of total flavonol glycosides in sample 2 raw materials, although meeting the requirement of the Pharmacopoeia that the content of total flavonol glycosides is greater than or equal to 0.40%, does not meet the requirement established in the present description that the content of total flavonol glycosides is greater than or equal to 0 .85%. The content of total flavonol glycosides in the raw material of sample 1 is not only greater than or equal to 0.40%, but also greater than or equal to 0.85%. The sample 1 raw material has a higher content of total flavonol glycosides and therefore is of better quality.

[00189] The results of the determination show that the content of terpene lactones in the raw material sample 1 and raw material sample 2 is 0.62% and 0.33%, respectively. The content of terpene lactones in sample 2 raw materials, although meeting the requirement of the Pharmacopoeia that the content of terpene lactones is greater than or equal to 0.25%, does not meet the requirement established in the present description that the content of terpene lactones is greater than or equal to 0 .40%. The content of terpene lactones in sample 1 raw materials is not only greater than or equal to 0.25%, but also greater than or equal to 0.40%. Sample 1 raw material has a higher content of terpene lactones, therefore, has a better raw material quality.

[00190] The results of the determination show that the area ratios of the chromatographic peaks of quercetin to kaempferide in the Sample 1 feed and Sample 2 feed are 0.87 and 0.55, respectively. Sample 1 feed is in the range of 0.65-1.2 specified in the present description, while sample 2 feed is not in the range of 0.65-1.2 specified in the present description. The area ratio of the chromatographic peaks of isorhamnetin to quercetin in the raw sample 1 and raw sample 2 is 0.21 and 0.14, respectively. Sample 1 raw material meets the requirement greater than 0.15 specified in the present description, while the raw material sample 2 does not satisfy the requirement greater than 0.15 specified in the present description. From the above proportional relationship, it can be seen that as a raw material, the raw material quality of sample 1 is better than the raw material quality of sample 2.

[00191] The determination results show that the content of sulfur dioxide residues in the Sample 1 feed and Sample 2 feed is 11 mg/kg and 88 mg/kg, respectively, both of which are less than or equal to 150 mg/kg. The content of sulfur dioxide residues in the feed sample 1 and feed sample 2 is within the established limits. Thus, the quality of sample 1 raw material and sample 2 raw material meets the requirements. However, sample 1 raw material is of better quality due to the lower content of sulfur dioxide residue.

[00192] At the same time, the determination results show that the total ash content of sample 1 feed and sample 2 feed is 3.8% and 6.9%, respectively, both of which are less than or equal to 10.0%. The content of acid-insoluble ash in the Sample 1 feed and Sample 2 feed is 0.68% and 1.54%, respectively, both of which are less than or equal to 2.0%. The extract content of sample 1 feed and sample 2 feed is 44.1% and 28.2%, respectively, both of which are greater than or equal to 25.0%. The impurity content of sample 1 feed and sample 2 feed is 0.57% and 1.33%, respectively, both of which are less than or equal to 2.0%. The water content of sample 1 feed and sample 2 feed is 5.3% and 10.1%, respectively, both of which are less than or equal to 12.0%. It can be seen that all of the above contents in the Sample 1 raw material and the Sample 2 raw material are within the specified range, and the quality of the raw materials is in accordance with the requirements. However, the respective contents of sample 1 raw materials are better than those of sample 2 raw materials. Therefore, the quality of sample 1 raw materials is better. Using sample 1 raw material, higher quality ginkgo biloba ketone ester raw materials can be obtained, thus higher quality granular ginkgo biloba ketone ester samples can be obtained.

[00193] As disclosed above, the present disclosure provides a granulated ginkgo biloba ketone ester and a process for making the same. Thanks to optimized production steps and conditions, high quality Ginkgo Biloba granular ketone ester can be obtained. Compared with the granular ginkgo biloba ketone ester obtained by conventional methods, the granular ginkgo biloba ketone ester of the present invention has a stable quality and a low quality loss rate. The uniformity of different batches of Ginkgo biloba granular ketone ester is good. In particular, the granulated ginkgo biloba ketone ester can be effectively controlled for ginkgolic acids by preventing the use of raw materials with a high content of ginkgolic acid in the granulated ginkgo biloba ketone ester, so as to effectively improve the safety of the granulated ginkgo biloba ketone ester. At the same time, the granular ginkgo biloba ketone ester obtained by the method of the present invention is excellent in taste, bitterness, stability, high temperature resistance, production characteristics and other characteristics. Therefore, the present invention effectively overcomes various shortcomings of the existing technology and is highly valuable for industrial applications.

[00194] The above embodiments only illustrate the principle and effects of the present disclosure, and do not limit the present disclosure. Modifications or variations of the embodiments described above may be made by those skilled in the art without departing from the spirit and scope of the disclosure. Therefore, all equivalent modifications or changes made by those skilled in the art without deviating from the spirit and technical concept disclosed in this disclosure should continue to be covered by the claims of the present invention.

Claims (48)

1. A method for obtaining a granular composition from an extract of ginkgo biloba leaves, including the following steps: A) mixing and agitating ginkgo biloba leaf extract, dextrin, sucrose powder, starch, low-substituted hydroxypropyl cellulose and aspartame according to the composition ratio and adding ethanol aqueous solution at the same time to obtain a soft material; C) obtaining wet granules from a soft material, followed by drying the wet granules; and C) sieving and straightening the dried granules, mixing, discharging and bagging to obtain a granular composition from the Ginkgo biloba leaf extract; where the ratio in the composition in mass parts includes the following: 35-45 parts of ginkgo biloba leaf extract; 450-500 parts of dextrin; 420-450 parts of sucrose powder; 10-30 parts of starch; 10-30 parts of low-substituted hydroxypropylcellulose; and 5-10 parts of aspartame; wherein the extraction to obtain the ginkgo biloba leaf extract includes the following steps: a) grinding ginkgo biloba leaves to obtain crushed ginkgo biloba leaves; b) adding an aqueous solution of ethanol to the crushed leaves of ginkgo biloba, performing a double extraction by heating under reflux and obtaining a first filtrate by filtration; c) adding water to the filtered drug residue remaining after filtration, performing a single extraction by heating under reflux, and then obtaining a second filtrate by filtration; d) combining and concentrating the second filtrate and the first filtrate into a thick paste, dissolving in purified water, cooling and filtering to obtain a filtrate; e) loading the filtrate into a macroporous resin column, sequentially eluting with 18% ethanol aqueous solution, 30% ethanol aqueous solution and 50% ethanol aqueous solution to obtain 18% ethanol eluent, 30% ethanol eluent and 50% ethanol eluent, respectively, which are first eluents; in the first eluents, the 50% ethanol eluent is the first high solvent eluent, and the combined solution of 18% ethanol eluent and 30% ethanol eluent is the first low solvent eluent; f) concentrating the first eluent with a low solvent concentration so that there is no smell of alcohol, loading the first eluent with a low solvent concentration into a polyamide column, and then eluting with ethanol to obtain an ethanol eluate, which is the second eluate; g) combining and concentrating the second eluent and the first eluent with a high concentration of solvent until the smell of alcohol disappears; h) extracting the concentrate with cyclohexane, discarding the cyclohexane extract, spray drying the concentrated solution, or concentrating the concentrated solution into a thick paste, and vacuum drying the thick paste; grinding dried products to obtain an extract of ginkgo biloba leaves; and where the granular formulation of ginkgo biloba leaf extract, based on the weight of the load of each package of 1 g, satisfies the following conditions: 1) the content of rutin is less than or equal to 1.60 mg/pack; 2) the content of quercetin is less than or equal to 0.160 mg/pack; 3) the content of bilobalide is 1.04-2.08 mg/package; 4) the content of ginkgolide J is 0.04-0.20 mg/pack; 5) the residual ethanol content is less than or equal to 0.2 mg/pack; 6) the content of biflavonoids is less than or equal to 0.008 mg/package, and these biflavonoids include amentoflavone, bilobetin and ginkgetin; 7) the content of genistin is 0 and the content of ginkgolide M is 0; 8) the content of terpene lactones is 2.4-4.8 mg/pack; 9) the content of total flavonoids is 14.08-26.40 mg/pack; and 10) the content of total flavonol glycosides is 9.6-14.0 mg/pack. 2. A process for preparing a granular formulation from ginkgo biloba leaf extract according to claim 1, wherein step A) comprises one or more of the following: A1) sucrose powder is obtained by grinding sucrose and sifting; A2) the starch is corn starch; A3) mixing and mixing time is 15-20 minutes; A4) an aqueous solution of ethanol has a mass percentage of 69-71%. 3. A method for preparing a granular formulation from ginkgo biloba leaf extract according to claim 1, wherein step B) comprises one or more of the following: B1) wet granules are obtained after sieving; B2) The drying oscillation time is 80-100 seconds, and the frequency is 5-8 times; B3) the drying stop temperature of the material is 66-70°C; B4) Drying time is 15-30 minutes. 4. A process for preparing a granular formulation from ginkgo biloba leaf extract according to claim 1, wherein step C) comprises one or more of the following: C1) sieving and straightening screen number is 12-40 mesh; C2) the stirring speed is 20-25 rpm; C3) mixing time is 5-6 minutes. 5. Granular composition from ginkgo biloba leaf extract, providing anti-PAF (platelet activating factor) effect and nerve nutrition, obtained by the method according to claim 1, where the content of total ginkgolic acids in the granular composition from ginkgo biloba leaf extract is less than or equal to 2 μg / package; where the total ginkgolic acids in the granular formulation from Ginkgo biloba leaf extract is calculated from the total content of Ginkgolic acid C13:0, Ginkgolic acid C15:1 and Ginkgolic acid C17:1. 6. Granular composition from the extract of ginkgo biloba leaves, providing an anti-PAF effect and nourishing the nerves, obtained by the method according to claim 1, where the ratio of the areas of chromatographic peaks of quercetin to kaempferide in the granular composition from the extract of ginkgo biloba leaves is 0.8-1.2 , and the ratio of the areas of the chromatographic peaks of isorhamnetin to quercetin in the granular composition from the extract of the leaves of ginkgo biloba is greater than 0.15. 7. Application of the method according to any one of paragraphs. 1-4 to obtain a granular formulation of ginkgo biloba leaf extract providing an anti-PAF effect and nerve nourishment.

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