Quality detection method and application of ginkgo and American ginseng preparation

Abstract

The invention provides a quality detection method and application of a ginkgo and American ginseng preparation, and relates to the technical field of pharmaceutical analysis. The quality detection method comprises the following steps: taking the gingko and American ginseng preparation, crushing, sieving, weighing, adding a solvent for extraction, filtering by a filter membrane, evaporating filtrate to dryness, adding methanol into residues for dissolution, and fixing the volume to obtain a test solution; the solvent is a mixed solution of n-butyl alcohol, methanol and acetic ether; the method comprises the following steps: weighing reference substances including bilobalide, bilobalide A, bilobalide B, bilobalide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1, and mixing the reference substances with a solvent to obtain a reference substance solution; determining the reference substance solution and the test solution by using a liquid chromatograph to obtain a chromatogram; the quality detection method has the advantages of being simple, rapid, stable, reliable, high in precision, good in reproducibility, easy to master and the like, and the saponin, the lactone and other components can be detected at the same time.

Description

Technical Field

[0001] The invention relates to the technical field of drug analysis, and in particular to a quality detection method and application of a ginkgo and ginseng preparation. Background Art

[0002] The main raw materials of Ginkgo Biloba and American Ginseng preparations are Ginkgo biloba leaves and American ginseng, which have the effects of invigorating qi and promoting blood circulation, widening the chest and unblocking the meridians. It is used as an auxiliary treatment for coronary heart disease (chest obstruction) with qi deficiency and blood stasis syndrome, and has an improving effect on symptoms such as chest tightness, shortness of breath, and palpitations.

[0003] American ginseng (Panaxquinquefolium L.) contains saponins, nucleotides, active polysaccharides, proteins, amino acids, inorganic elements and flavonoids, and has multiple pharmacological effects such as immunomodulation, anti-depression, anti-tumor, anti-inflammatory, antioxidant and hypolipidemic. The quality of American ginseng medicinal materials varies greatly, which brings hidden dangers to the safe use of drugs in clinical practice. The quality of raw materials greatly determines the quality of their preparations and health foods. The quality inspection and control of raw materials are of great significance to ensure clinical efficacy.

[0004] Ginkgo biloba is the dried leaves of Ginkgo biloba, a plant of the Ginkgoaceae family. It is mainly used to treat cardiovascular and cerebrovascular diseases and nervous system diseases. The terpenelactones and flavonoids contained in Ginkgo biloba are the main medicinal active ingredients of Ginkgo biloba.

[0005] Chinese patent CN102309531B discloses a method for determining the fingerprint of American ginseng medicinal material. The method uses high performance liquid chromatography to determine its fingerprint and uses an evaporative light scattering detector for detection. The chromatographic conditions of the high performance liquid chromatography are: the chromatographic column is a C18 column; the mobile phase: the mobile phase A is acetonitrile, the mobile phase B is a 0.05% acetic acid aqueous solution, and the gradient elution is performed.

[0006] The mobile phase gradient was as follows:

[0007] 0→20 minutes, mobile phase A is uniformly reduced from 80% to 79%, 20→25 minutes, mobile phase A is uniformly reduced from 79% to 74%, 25→29 minutes, mobile phase A is uniformly reduced from 74% to 73%, 29→43 minutes, mobile phase A is uniformly reduced from 73% to 66%, 43→47 minutes, mobile phase A is uniformly reduced from 66% to 64%, 47→54 minutes, mobile phase A is uniformly reduced from 64% to 57%, 54→55 minutes, mobile phase A is uniformly reduced from 57% to 55%, 55→59 minutes, mobile phase A is uniformly reduced from 55% to 50%, 59→65 minutes, mobile phase A is uniformly reduced from 50% to 5%, 65→69 minutes, mobile phase A is kept at 5%. However, this scheme is only for the quality control of the active ingredients of American ginseng.

[0008] Chinese patent CN 115112783 A provides a fingerprint detection method for ginkgo leaves, comprising the following steps: taking a ginkgo leaf reference substance and a test substance for detection, and the chromatographic conditions of the detection include: a chromatographic column of ZORBAX SB-C18 (4.6×250 mm, 5 μm); a mobile phase of 0.1 volume % formic acid water (A)-methanol (B); a gradient elution program of: 0-10 min: 90 volume %-80 volume % A; 10-25 min: 80 volume %-70 volume % A; 25-35 min: 70 volume % A; 35-45 min: 70 volume %-58 volume % A; 45-70 min: 58 volume %-45 volume % A; 70-85 min: 45 volume %-30 volume % A; 85-95 min: 30 volume %-5 volume % A; 95-115 min: 5 volume % A. Flow rate 0.9-1.1mL / min; detection wavelength 360nm, column temperature 25-35℃, injection volume 10μL; evaporative light scattering detector parameters: drift tube temperature 110℃, carrier gas flow rate 3.0mL / min, gain value 4. It provides a reliable analytical method for the effective control of the quality of Ginkgo biloba products. However, this scheme only controls the quality of the active ingredients in Ginkgo biloba.

[0009] Traditional Chinese medicine fingerprint has the characteristics of overall, macroscopic, and fuzzy analysis. It can achieve the purpose of overall quality control by describing the overall characteristics of traditional Chinese medicine and using appropriate fuzzy processing methods. Therefore, it has become an effective means of quality control of traditional Chinese medicine. Among them, chromatographic fingerprint analysis can visualize the overall characteristics of various chemical components contained in traditional Chinese medicine, thereby revealing quality problems that are difficult to find through conventional inspections. However, there is currently no quality control method for the active ingredients of ginkgo and ginseng preparations. Summary of the invention

[0010] In view of the above problems, the present invention provides a quality detection method for a Ginkgo and Western ginseng preparation. By optimizing the sample pretreatment method, elution solvent and elution procedure, the separation and detection of active ingredients in the Ginkgo and Western ginseng preparation can be achieved, and a more comprehensive and accurate evaluation of the quality of the Ginkgo and Western ginseng preparation can be made.

[0011] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0012] In one aspect, the present invention provides a method for detecting the quality of a Ginkgo biloba preparation, comprising the following steps:

[0013] S1: Preparation of the test solution: Ginkgo biloba and American ginseng preparation is crushed, sieved, weighed, extracted with solvent, filtered with filter membrane, the filtrate is evaporated to dryness, the residue is dissolved with methanol, and the volume is fixed to obtain the test solution; the solvent is a mixed solution of n-butanol, methanol and ethyl acetate;

[0014] S2: Preparation of reference solution: weigh reference substances bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1, and mix with solvent to obtain reference solution;

[0015] S3: Use a liquid chromatograph to measure the reference solution and the test solution to obtain a chromatogram;

[0016] The chromatographic conditions are:

[0017] Chromatographic column: non-polar reverse phase chromatography column;

[0018] Mobile Phase:

[0019] Mobile phase A: acetonitrile and chloroform;

[0020] Mobile phase B: 0.15-0.3wt% phosphoric acid solution and 0.01-0.015wt% acetic acid solution;

[0021] Flow rate: 0.8-1.2 mL min -1 ;

[0022] Column temperature: 30℃-40℃;

[0023] The elution method is gradient elution, and the elution gradient is:

[0024] 0-5min, the volume ratio of mobile phase A:mobile phase B was 8-10:90-92;

[0025] 5-15min, the volume ratio of mobile phase A:mobile phase B is 8-18:82-92;

[0026] 15-20min, the volume ratio of mobile phase A:mobile phase B is 10-28:72-90;

[0027] 20-50min, the volume ratio of mobile phase A:mobile phase B is 23-35:65-77;

[0028] 50-70min, the volume ratio of mobile phase A:mobile phase B is 25-80:20-75.

[0029] Preferably, in S1, the sieving is through a 40-60 mesh sieve; further preferably, in S1, the sieving is through a 40 mesh sieve.

[0030] Preferably, in S1, the solid-liquid ratio of the Ginkgo biloba and American ginseng preparation to the solvent is 1:10-20 g / mL; further preferably, in S1, the solid-liquid ratio of the Ginkgo biloba and American ginseng preparation to the solvent is 1:14 g / mL.

[0031] Preferably, in S1, the extraction is ultrasonic extraction;

[0032] Preferably, the parameters of the ultrasonic extraction are: ultrasonic power of 300-550W, frequency of 30-40kHz, and time of 20-45min; further preferably, the parameters of the ultrasonic extraction are: ultrasonic power of 420W, frequency of 35kHz, and time of 35min.

[0033] Preferably, in S1, the pore size of the filter membrane is 0.2-0.35 μm, and further preferably, in S1, the pore size of the filter membrane is 0.24 μm.

[0034] Preferably, in S1, the concentration of the test solution is 1 g / 50 mL;

[0035] Preferably, in S1, in the mixed solution of n-butanol, methanol and ethyl acetate, methanol is a methanol solution with a volume fraction of 70-85%; further preferably, in S1, in the mixed solution of n-butanol, methanol and ethyl acetate, methanol is a methanol solution with a volume fraction of 85%.

[0036] Preferably, in S1, the volume ratio of n-butanol, methanol and ethyl acetate is 8-12:5-7.5:0.5-1.5; further preferably, in S1, the volume ratio of n-butanol, methanol and ethyl acetate is 10:6.5:1.

[0037] Preferably, in S2, the concentration of each reference substance in the reference solution is: bilobalide 10-14 μg / mL, ginkgolide A 6-14 μg / mL, ginkgolide B 5-12 μg / mL, ginkgolide C 1-160 μg / mL, ginsenoside F11 7-15 μg / mL, ginsenoside Rb1 80-130 μg / mL, ginsenoside Re 50-80 μg / mL and ginsenoside Rg1 1-10 μg / mL; further preferably, in S2, the concentration of each reference substance in the reference solution is: bilobalide 12 μg / mL, ginkgolide A 8 μg / mL, ginkgolide B 7 μg / mL, ginkgolide C 6 μg / mL, ginsenoside F11 7-15 μg / mL, ginsenoside Rb1 80-130 μg / mL, ginsenoside Re 50-80 μg / mL and ginsenoside Rg1 1-10 μg / mL. 12μg / mL, ginsenoside Rb1120μg / mL, ginsenoside Re 60μg / mL, and ginsenoside Rg1 5μg / mL;

[0038] Preferably, in S2, the solvent is a mixture of 85% by volume methanol solution and acetonitrile;

[0039] Preferably, the volume ratio of the methanol solution with a volume fraction of 85% to acetonitrile is 8-12:1; further preferably, the volume ratio of the methanol solution with a volume fraction of 85% to acetonitrile is 10:1.

[0040] Preferably, in S3, the chromatographic column is a non-polar reverse phase chromatographic column using octadecylsilane bonded silica gel as a filler;

[0041] Preferably, in S3, in the mobile phase A, the volume ratio of acetonitrile to chloroform is 8-12:1; further preferably, in S3, in the mobile phase A, the volume ratio of acetonitrile to chloroform is 9:1.

[0042] Preferably, in S3, mobile phase B is: 0.25wt% phosphoric acid solution and 0.01wt% acetic acid solution;

[0043] Preferably, in S3, the volume ratio of 0.25wt% phosphoric acid solution to 0.01wt% acetic acid solution in mobile phase B is 3-5:1; further preferably, in S3, the volume ratio of 0.25wt% phosphoric acid solution to 0.01wt% acetic acid solution in mobile phase B is 4:1.

[0044] Preferably, in S3, the flow rate is 1 mL min -1 ;

[0045] Preferably, in S3, the column temperature is 35°C;

[0046] Preferably, in S3, the detection wavelength of the chromatographic conditions is 280-310 nm.

[0047] Preferably, in S3, the elution gradient is:

[0048] Time / min Mobile phase A / Vt% Mobile phase B / Vt% 0 10 90 5 10 90 15 14 86 20 25 75 35 30 70 50 30 70 60 78 22 70 78 22

[0049] On the other hand, the present invention provides application of the above-mentioned quality detection method of ginkgo and American ginseng preparation in quality control of ginkgo and American ginseng preparation using ginkgo leaves and American ginseng as raw materials.

[0050] Preferably, the ginkgo and American ginseng preparation comprises ginkgo and American ginseng capsules and ginkgo and American ginseng granules.

[0051] Compared with the prior art, the present invention has the following beneficial effects:

[0052] 1. The technical solution of the present invention obtains more comprehensive information on the active ingredients by optimizing the extraction solvent and extraction time to ensure that the active ingredients in the Ginkgo and American ginseng preparations are extracted to the greatest extent; by optimizing the selection of mobile phase, mobile phase elution program, chromatographic column and other conditions, it ensures that the active ingredients in the Ginkgo and American ginseng preparations are well separated, with good peak shape and no interference.

[0053] 2. The quality determination method of the ginkgo and American ginseng preparation provided by the present invention overcomes the defect that the quality of the ginkgo and American ginseng preparation cannot be comprehensively and clearly detected due to the interference caused by the complex chemical components of traditional Chinese medicine under preferred chromatographic conditions.

[0054] 3. This quality detection method has the advantages of being simple and fast, stable and reliable, highly precise, reproducible, and easy to master. DETAILED DESCRIPTION

[0055] In order to make the technical means, creative features, purpose and effect of the present invention easy to understand, the present invention is further explained below in conjunction with specific embodiments, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without making creative work all belong to the protection scope of the present invention. It is worth noting that the raw materials used in the present invention are all common commercial products, and their sources are not specifically limited. The technology and scientific terms used in the embodiments have the meanings commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0056] Instruments and reagents

[0057] Agilent 1260 InfinityⅡ liquid chromatograph (vacuum degasser, binary pump, autosampler, column oven, DAD detector);

[0058] Analytical balance, model: Mettler Toledo PL203 and XS205;

[0059] KQ-300DB CNC ultrasonic cleaner (300W, 40kHz).

[0060] Phosphoric acid, manufacturer: Sinopharm Chemical Reagent Co., Ltd.;

[0061] Acetic acid, manufacturer: Sinopharm Chemical Reagent Co., Ltd.;

[0062] Methanol, manufacturer: Sinopharm Chemical Reagent Co., Ltd.;

[0063] Acetonitrile, manufacturer: Sinopharm Chemical Reagent Co., Ltd.;

[0064] n-Butanol, manufacturer: Sinopharm Chemical Reagent Co., Ltd.;

[0065] Ethyl acetate, manufacturer: Sinopharm Chemical Reagent Co., Ltd.;

[0066] Water, purified water.

[0067] Sample: Ginkgo and Panax ginseng preparation (produced by Guangxi Liangmianzhen Yikan Pharmaceutical Co., Ltd.).

[0068] Reference substances: bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1 were all purchased from China Food and Drug Inspection Institutes. The purity of other reagents was required to be analytical grade.

[0069] Example 1

[0070] A quality detection method for a ginkgo and ginseng preparation, specifically comprising:

[0071] S1: Preparation of test solution: Ginkgo biloba and American ginseng preparation is crushed, passed through a 40-mesh sieve, weighed, and extracted with a solvent, wherein the solid-liquid ratio of the Ginkgo biloba and American ginseng preparation to the solvent is 1:14 g / mL, filtered with a 0.24 μm filter membrane, the filtrate is evaporated to dryness, the residue is dissolved in methanol, and the volume is fixed to obtain a test solution; the concentration of the test solution is 1 g / 50 mL;

[0072] The solvent is a mixed solution of n-butanol, 85% volume fraction of methanol and ethyl acetate; the volume ratio of n-butanol, 85% volume fraction of methanol and ethyl acetate is 10:6.5:1.

[0073] S2: Preparation of reference solution: weigh reference substances bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1, and mix them with a solvent to obtain a reference solution; the solvent is a mixture of 85% by volume methanol solution and acetonitrile; the volume ratio of the 85% by volume methanol solution to acetonitrile is 10:1.

[0074] The concentration of each reference substance in the reference solution is: bilobalide 8 μg / mL, ginkgolide A 12 μg / mL, ginkgolide B 10 μg / mL, ginkgolide C 14 μg / mL, ginsenoside F11 8 μg / mL, ginsenoside Rb1 10 μg / mL, ginsenoside Re 10 μg / mL and ginsenoside Rg1 14 μg / mL;

[0075] S3: Use a liquid chromatograph to measure the reference solution and the test solution to obtain a chromatogram;

[0076] The chromatographic conditions are:

[0077] Chromatographic column: USHA C18, size 2.1mm×50mm, filler particle size 1.8μm;

[0078] Mobile Phase:

[0079] Mobile phase A: a mixed solution of acetonitrile and chloroform, with a volume ratio of 9:1;

[0080] Mobile phase B: a mixed solution of 0.25wt% phosphoric acid solution and 0.01wt% acetic acid solution, with a volume ratio of 4:1;

[0081] Flow rate: 1 mL min -1 ;

[0082] Column temperature: 35°C;

[0083] Injection volume: 10 μL;

[0084] Detection wavelength: 290nm;

[0085] The elution method is gradient elution, and the elution gradient is:

[0086] Table 1. Elution gradient

[0087] Time / min Mobile phase A / Vt% Mobile phase B / Vt% 0 10 90 5 10 90 15 14 86 20 25 75 35 30 70 50 30 70 60 78 22 70 78 22

[0088] The information of the obtained spectrum is shown in Table 2 below:

[0089] Table 2. Spectral information

[0090] Sample name Peak Retention time / min Peak area Bilobalide 1 22.821 13245 Ginsenoside Rg1 2 24.315 8045 Ginsenoside Re 3 28.059 53147 Ginkgolide C 4 32.078 9683 Ginsenoside F11 5 35.278 11257 Ginsenoside Rb1 6 42.182 103284 Ginkgolide A 7 44.239 10598 Ginkgolide B 8 46.855 9178

[0091] Example 2

[0092] The difference from Example 1 is that the detection wavelengths are 280nm, 300nm, and 310nm, respectively, and the rest are the same. The results show that the characteristic peaks of the Ginkgo biloba preparation can also be separated at the detection wavelengths of 280nm, 300nm, and 310nm, but the peaks are more complete and the characteristic peaks are obvious at the detection wavelength of 290nm.

[0093] Example 3

[0094] The difference from Example 1 is that the experiments were conducted at column temperatures of 30°C and 40°C, respectively, and the rest were the same. The results showed that the separation of the main chromatographic peaks was the best at 35°C, and the peak shape was the best.

[0095] Example 4

[0096] The difference from Example 1 is that 0.8 mL·min -1 、0.9mL·min -1 、1.1mL·min -1 、1.2mL·min -1 The results show that the flow rate is 1.0 mL min -1 When the main chromatographic peaks are separated best, the peak shape is the best.

[0097] Comparative Example 1-Comparative Example 6

[0098] Different from Example 1, only the solvent in S1 was changed in Comparative Examples 1 to Comparative Examples 6, and the other steps were the same. The content determination results are shown in Table 3 below:

[0099] Table 3. Effect of extraction solvent on the content of each component

[0100]

[0101]

[0102] Comparative Example 7

[0103] Compared with Example 1, only the gradient elution procedure was changed, and the rest were the same:

[0104] The gradient elution program is as follows:

[0105] Table 4. Elution gradient

[0106] Time / min Mobile phase A / Vt% Mobile phase B / Vt% 0 25 75 15 35 65 40 45 55 50 50 50 70 60 60

[0107] Comparative Example 8

[0108] Compared with Example 1, only the mobile phase A was changed to acetonitrile, and the rest were the same.

[0109] Comparative Example 9

[0110] Compared with Example 1, only the mobile phase A was changed to chloroform, and the rest were the same.

[0111] Comparative Example 10

[0112] Compared with Example 1, only the mobile phase A was changed to methanol, and the rest were the same.

[0113] Comparative Example 11

[0114] Compared with Example 1, only the mobile phase B was changed to 0.25 wt % phosphoric acid solution, and the rest were the same.

[0115] Comparative Example 12

[0116] Compared with Example 1, only the mobile phase B was changed to 0.01 wt % sulfuric acid solution, and the rest were the same.

[0117] Test example

[0118] According to the detection method of Example 1, the specificity, linearity, precision, stability, repeatability and detection limit were investigated.

[0119] 1. Exclusivity

[0120] Accurately measure 10 μL of blank solvent (the solvent is a mixed solution of n-butanol, 85% volume fraction of methanol and ethyl acetate; the volume ratio of n-butanol, 85% volume fraction of methanol and ethyl acetate is 10:6.5:1) and the test solution, respectively, inject them into liquid chromatograph, and determine to obtain the result.

[0121] Result: The blank solvent had no interference with the test product.

[0122] 2. Linear relationship

[0123] The reference substances bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1 were accurately weighed and diluted with solvent to prepare a reference substance mixed solution with a certain concentration gradient; the solvent was a mixture of 85% methanol solution and acetonitrile; the volume ratio of 85% methanol solution and acetonitrile was 10:1. 10 μL of the reference substance mixed solution from low to high concentration was injected respectively, and the linear relationship between the concentration (μg / mL) of each component and the corresponding peak area was investigated. Results The reference substances bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1 had a good linear relationship between the concentration and the corresponding area, and the linear range of the injection amount of each component met the detection requirements. Correlation coefficient: R 2 =0.9999.

[0124] 3. Precision

[0125] Take the same portion of Ginkgo biloba preparation to make a test solution with a concentration of 1g / 50mL, inject 6 times continuously, 210μL each time, record the retention time and peak area of ​​peaks 1, 2, 3, 4, 5, 6, 7, and 8 respectively, and calculate the relative standard deviation RSD of the retention time and peak area of ​​each peak. The RSD of the retention time of each peak is 0.08%-0.15%, and the RSD of the peak area is 0.11%-0.25%, both less than 3%, indicating that the instrument has good precision. The calculation results are shown in Table 5.

[0126] Table 5. Precision results

[0127]

[0128]

[0129] 4. Stability

[0130] According to the test sample preparation method of Example 1, the retention time of the same sample was measured at 0, 4, 8, 12, 36, and 48 hours, and the RSD% was 0.18%-0.86%, indicating that the sample solution had good stability within 48 hours. The results are shown in Table 6 below.

[0131] Table 6. Stability results

[0132]

[0133] 5. Repeatability

[0134] According to the test sample preparation method of Example 1, 6 test sample solutions were prepared in parallel for the same batch of samples, and the content was determined. The RSD% was 0.45%-0.92%, indicating that the method had good repeatability. The results are shown in Table 7 below.

[0135] Table 7. Repeatability results

[0136]

[0137] According to the detection method in the detection example, the linear correlation coefficient, precision, stability and repeatability of Example 1 and Comparative Examples 7-12 were tested, and the results are shown in Tables 8-11 below.

[0138] Table 8. Linear correlation coefficient R of Example 1 and Comparative Examples 7-12 2 contrast(%)

[0139] Element Example 1 Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 Comparative Example 12 Bilobalide 0.9999 0.9991 0.9992 0.9991 0.9990 0.9990 0.9990 Ginsenoside Rg1 0.9999 0.9991 0.9990 0.9990 0.9993 0.9991 0.9995 Ginsenoside Re 0.9999 0.9994 0.9991 0.9992 0.9991 0.9990 0.9991 Ginkgolide C 0.9999 0.9992 0.9991 0.9992 0.9991 0.9990 0.9990 Ginsenoside F11 0.9999 0.9990 0.9991 0.9994 0.9990 0.9991 0.9992 Ginsenoside Rb1 0.9999 0.9991 0.9992 0.9990 0.9994 0.9991 0.9990 Ginkgolide A 0.9999 0.9992 0.9992 0.9991 0.9990 0.9991 0.9990 Ginkgolide B 0.9999 0.9990 0.9992 0.9991 0.9990 0.9991 0.9990

[0140] Table 9. Comparison of precision RSD (peak area) of Example 1 and Comparative Examples 7-12 (%)

[0141]

[0142]

[0143] Table 10. Comparison of stability RSD (retention time) of Example 1 and Comparative Examples 7-12 (%)

[0144] Element Example 1 Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 Comparative Example 12 Bilobalide 0.21 3.4 3.1 2.9 2.8 2.1 2.7 Ginsenoside Rg1 0.24 3.8 3.4 3.2 2.9 3.0 3.3 Ginsenoside Re 0.56 3.2 3.5 3.4 4.0 4.2 3.1 Ginkgolide C 0.86 5.1 5.5 5.3 4.9 5.0 4.8 Ginsenoside F11 0.18 2.1 2.2 2.5 1.9 2.0 2.4 Ginsenoside Rb1 0.41 3.1 2.8 2.9 2.5 2.4 2.7 Ginkgolide A 0.18 2.5 2.1 2.4 2.0 2.9 2.5 Ginkgolide B 0.81 4.9 4.8 4.5 3.6 3.5 4.0

[0145] Table 11. Comparison of repeatability RSD (peak area) of Example 1 and Comparative Examples 7-12 (%)

[0146]

[0147]

[0148] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A method for detecting the quality of a ginkgo and ginseng preparation, characterized in that: The following steps are involved: S1: Preparation of the test solution: Ginkgo biloba and American ginseng preparation is crushed, sieved, weighed, extracted with solvent, filtered with filter membrane, the filtrate is evaporated to dryness, the residue is dissolved with methanol, and the volume is fixed to obtain the test solution; the solvent is a mixed solution of n-butanol, methanol and ethyl acetate; S2: Preparation of reference solution: weigh reference substances bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginsenoside F11, ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1, and mix with solvent to obtain reference solution; S3: Use a liquid chromatograph to measure the reference solution and the test solution to obtain a chromatogram; The chromatographic conditions are: Chromatographic column: non-polar reverse phase chromatography column; Mobile Phase: Mobile phase A: acetonitrile and chloroform; Mobile phase B: 0.15-0.3wt% phosphoric acid solution and 0.01-0.015wt% acetic acid solution; Flow rate: 0.8-1.2 mL min -1 ; Column temperature: 30℃-40℃; The elution method is gradient elution, and the elution gradient is: 0-5min, the volume ratio of mobile phase A:mobile phase B was 8-10:90-92; 5-15min, the volume ratio of mobile phase A:mobile phase B is 8-18:82-92; 15-20min, the volume ratio of mobile phase A:mobile phase B is 10-28:72-90; 20-50min, the volume ratio of mobile phase A:mobile phase B is 23-35:65-77; 50-70min, the volume ratio of mobile phase A:mobile phase B is 25-80:20-75.

2. The quality inspection method according to claim 1, characterized in that: In S1, the solid-liquid ratio of the Ginkgo biloba preparation and the solvent is 1:10-20g / mL; the extraction is ultrasonic extraction; the parameters of the ultrasonic extraction are: the ultrasonic power is 300-550W, the frequency is 30-40kHz, and the time is 20-45min; the pore size of the filter membrane is 0.2-0.35μm; the concentration of the test solution is 1g / 50mL; the volume ratio of n-butanol, methanol and ethyl acetate is 8-12:5-7.5:0.5-1.

5.

3. The quality inspection method according to claim 1, characterized in that: In S2, the concentration of each reference substance in the reference substance solution is: bilobalide 10-14μg / mL, ginkgolide A 6-14μg / mL, ginkgolide B 5-12μg / mL, ginkgolide C1-160μg / mL, ginsenoside F11 7-15μg / mL, ginsenoside Rb1 80-130μg / mL, ginsenoside Re 50-80μg / mL and ginsenoside Rg1 1-10μg / mL.

4. The quality inspection method according to claim 1, characterized in that: In S2, the solvent is a mixture of a methanol solution with a volume fraction of 85% and acetonitrile; the volume ratio of the methanol solution with a volume fraction of 85% and acetonitrile is 8-12:

1.

5. The quality inspection method according to claim 1, characterized in that: In S3, the chromatographic column is a non-polar reverse phase chromatographic column using octadecylsilane bonded silica gel as a filler.

6. The quality inspection method according to claim 1, characterized in that: In S3, the volume ratio of acetonitrile and chloroform in the mobile phase A is 8-12:1; the mobile phase B is: 0.25wt% phosphoric acid solution and 0.01wt% acetic acid solution; the volume ratio of 0.25wt% phosphoric acid solution and 0.01wt% acetic acid solution in the mobile phase B is 3-5:

1.

7. The quality inspection method according to claim 1, characterized in that: In S3, The flow rate is 1 mL min -1 ; The column temperature is 35°C; The detection wavelength of the chromatographic conditions is 280-310 nm.

8. The quality inspection method according to claim 1, characterized in that: In S3, the elution gradient is: 。 9. Application of the quality detection method of the ginkgo and American ginseng preparation according to any one of claims 1 to 8 in the quality control of the ginkgo and American ginseng preparation using ginkgo leaves and American ginseng as raw materials.

10. The use according to claim 9, characterized in that: The ginkgo and ginseng preparation comprises ginkgo and ginseng capsules and ginkgo and ginseng granules.

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