CN113533612A - Detection method for controlling quality of vitality securing granules - Google Patents

Abstract

The invention discloses a detection method for controlling the quality of vitality securing granules. It comprises the following steps: detecting whether the sample solid particles are qualified or not by thin-layer chromatography detection and characteristic spectrum identification; and then, determining the weight average molecular weight of the sample solid particles through the molecular weight and the molecular weight distribution so as to finally determine whether the sample solid particles are qualified. The method qualitatively controls the quality of the test sample and quantitatively controls the content of specific components in the test sample so as to ensure the stability and uniformity of the test sample; and further has the characteristics of simplicity and convenience and good reproducibility through a molecular weight and molecular weight distribution determination method.

Description

Detection method for controlling quality of vitality securing granules

Technical Field

The invention relates to a detection method for controlling the quality of vitality securing granules, belonging to the field of medicine quality control and management.

Background

The vitality securing granule is brown granule containing crude polysaccharide of radix astragali and rhizoma Phragmitis of sun-dried ginseng; sweet and bitter. The functional indications are as follows: to replenish qi and consolidate the constitution. Can be used for the adjuvant treatment of listlessness, immunologic hypofunction, and hematopoietic hypofunction caused by cancer chemotherapy and radiotherapy.

The existing quality detection method of the vitality securing granules comprises thin-layer identification (ginseng rhizome); the quality control method comprises the inspection item (conventional inspection method of granules) and the content measurement item (astragalus polysaccharide), but the quality control method is difficult to effectively control due to the lack of a highly specific inspection method.

Disclosure of Invention

The invention aims to provide a detection method for controlling the quality of a solid particle.

Aiming at the defects in the prior art, the invention improves the existing quality detection method of the vigor-strengthening granules, increases the variety of the ginsenoside by identifying the monomer saponin through a ginsenoside thin layer, and increases the characteristic spectrum of the ginsenoside and the molecular weight distribution of large molecular weight components in the ginsenoside.

The invention provides a detection method for controlling the quality of vitality securing granules, which comprises the following steps:

(1) and (3) thin-layer chromatography detection: 1) extracting sample solid powder to obtain sample methanol solution, and preparing reference methanol solutions from ginsenoside Rb1, ginsenoside Re, and ginsenoside Rg 1;

2) respectively measuring the methanol solution of the test sample and the methanol solution of the reference sample by adopting a thin-layer chromatography, and respectively displaying spots and fluorescent spots with the same color on corresponding positions of the two, so that the solid particles of the test sample are qualified; otherwise, the sample solid particles are unqualified;

(2) and (3) characteristic map identification: 1) treating the sample solid particle powder to prepare a sample solution a; preparing reference solutions from the ginsenoside Rg1 reference, the ginsenoside Re reference, the ginsenoside Rb1 reference, the ginsenoside Rb2 reference, the ginsenoside Rb3 reference, the ginsenoside Rc reference and the ginsenoside Rd reference, respectively; 2) respectively detecting the test solution a and the reference substance solution by liquid chromatography, wherein 7 characteristic peaks are presented in a map of the test solution a, the retention time of each characteristic peak is the same as that of the reference substance solution, and the test solid particles are qualified; otherwise, the sample solid particles are unqualified;

the conditions of the liquid chromatography were as follows: octadecylsilane chemically bonded silica is used as a filling agent;

acetonitrile is taken as a mobile phase A, and 0.1% phosphoric acid is taken as a mobile phase B;

flow rate: 1.0 mL/min; detection wavelength: 203 nm;

gradient elution was performed as follows in volume percent: 0min, mobile phase A18%, mobile phase B82%; 40min, mobile phase A18%, mobile phase B82%; 50min, mobile phase A22%, mobile phase B78%; 70min, mobile phase A28%, mobile phase B72%; 100min, mobile phase A38%, mobile phase B62%; 110min, mobile phase A18%, mobile phase B82%;

(3) determination of molecular weight and molecular weight distribution: 1) adding mobile phase into dextran-1000, dextran-5000, dextran-12000, dextran-25000, dextran-50000 and anhydrous glucose with known molecular weight to obtain solutions each containing 5mg per 1ml as reference solution b;

taking 10mg of the test sample solid particle powder qualified by the steps (1) and (2) or the steps (2) and (1) in sequence, adding 2 mL0.7% of sodium sulfate for dissolution, and filtering by using a 0.45-micron microporous filter membrane to obtain a test sample solution b;

2) the reference substance solution b and the test substance solution b are measured by chromatography, chromatogram is recorded, and GPC polysaccharide special software is adopted for processing; when the weight average molecular weight of the sample solid particles is 3000-8000, the sample solid particles are qualified; otherwise, the sample solid particles are unqualified;

the chromatographic conditions were as follows: sepax SRT SEC-100(8.0 mm. times.300 mm) gel column (Seiki Ke Tech Co., Ltd., USA); 0.7% sodium sulfate as mobile phase; the column temperature is 35 ℃, and the flow rate is 0.5 mL/min; the number of theoretical plates, a differential refractometer detector, a software workstation dedicated to LZD GPC2010, should be no less than 3000 calculated as glucose peaks.

In the step (1) -1), the extraction process of the sample solid particles is as follows: taking the sample solid particle powder, counting according to 1g of ginseng head, adding 100ml of trichloromethane, heating and refluxing for 1 hour, removing chloroform liquid, volatilizing solvent from decoction dregs, adding 5ml of water, uniformly stirring and wetting, adding 100ml of water-saturated n-butyl alcohol, carrying out ultrasonic treatment for 30 minutes, absorbing supernatant, adding 2 times of ammonia sample, shaking uniformly, standing for layering, taking supernatant, evaporating to dryness, and adding 5ml of methanol to dissolve residues to obtain a sample solution;

the reference methanol solution is prepared by mixing ginsenoside Rb1 reference, ginsenoside Re reference, and ginsenoside Rg1 reference with methanol to obtain mixed solution containing 2mg of each 1 ml.

In the above method, the thin layer chromatography: sucking three kinds of control methanol solutions 5 μ l each, respectively dropping on the same silica gel G thin layer plate, developing with chloroform-ethyl acetate-water (15:40:22:10) lower layer solution at 10 deg.C below as developing agent, taking out, air drying, spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, and respectively inspecting under sunlight and ultraviolet lamp (365 nm).

In the above method (2) -1), the method for treating the sample solid particle powder is as follows:

taking 1g of the test sample solid particle powder passing through a No. four sieve, adding 50ml of methanol, and carrying out ultrasonic treatment for 30 minutes at the power of 250W and the frequency of kHz; filtering, measuring 25ml of subsequent filtrate, evaporating to dryness, dissolving the residue in 25ml of water-saturated n-butanol, placing in a separating funnel, adding 3 times of ammonia test solution, washing, shaking uniformly, standing for layering, taking n-butanol, evaporating to dryness, adding methanol solution into the residue, transferring to a 5ml measuring flask, adding methanol to dilute to a scale, shaking uniformly, filtering, and taking subsequent filtrate to obtain the sample solution a.

The invention has the following advantages:

1. the Astragalus polysaccharides as one of the effective components of the granule is water-soluble heteropolysaccharide obtained by extracting, concentrating and purifying dried root of Astragalus mongholicus (Fisch.) bge or Astragalus membranaceus (Fisch.) bge of Leguminosae. The molecular structure and molecular weight have close relationship with biological activity and purity, so that the research on molecular weight distribution is an important factor for controlling and improving the product quality, and the research and establishment of the molecular weight of astragalus polysaccharide and the molecular weight distribution standard thereof are necessary. The invention uses polysaccharide molecular weight and molecular weight distribution determination method to research and limit the molecular weight and distribution of astragalus polysaccharide. The method has the advantages of simplicity, convenience, good reproducibility and the like; and can make chemical detection and action of said medicine have a certain correlation.

2. The method adopts the characteristic spectrum of the traditional Chinese medicine for identification, adopts liquid chromatography to detect the processed test sample to obtain the characteristic spectrum, and analyzes to obtain 7 characteristic peaks capable of marking the characteristics of various groups in the test sample so as to qualitatively control the quality of the test sample; then, the content of specific components in the test sample is quantitatively controlled through the area and the proportion of main characteristic peaks of the characteristic map, so that the stability and the uniformity of the test sample are ensured, and the safety, the effectiveness and the controllability of the traditional Chinese medicine are ensured.

3. The invention establishes a quality control method of the Guyuan granules, which comprises thin layer identification, molecular weight distribution and a characteristic map, expresses the quality of finished products to achieve the purposes of controlling process operation and raw medicinal materials, achieves the purposes of operability and controllability, and improves the quality control level of the Guyuan granules.

Drawings

FIG. 1 shows the results of thin layer chromatography of 5 lots of samples and a reference sample according to the present invention, wherein 1 in FIG. 1 represents a granule for consolidating essence (lot No. 1), 2 represents a granule for consolidating essence (lot No. 2), 3 represents a granule for consolidating essence (lot No. 3), 4 represents a granule for consolidating essence (lot No. 5), 5 represents a reference drug, and 6 represents a mixed reference.

Fig. 2 shows thin layer chromatography results of 10 test samples and a control sample according to the present invention, wherein 1 in fig. 2 indicates a solid particle (batch 1), 2 indicates a solid particle (batch 2), 3 indicates a solid particle (batch 3), 4 indicates a solid particle (batch 4), 5 indicates a solid particle (batch 5), 6 indicates a solid particle (batch 6), 7 indicates a solid particle (batch 7), 8 indicates a solid particle (batch 8), 9 indicates a solid particle (batch 9), 10 indicates a solid particle (batch 10), 11 indicates a control drug, and 12 indicates a mixed control.

FIG. 3 is a chromatogram of a sample derived product, wherein peak 2 in FIG. 3 is rhamnose, peak 3(S) is glucose, peak 4 is galactose, and peak 5 is arabinose.

FIG. 4 is a chromatogram of a derivative product of a mixed reference peak, in FIG. 4, peak 2 is rhamnose, peak 3(S) is glucose, peak 4 is galactose, and peak 5 is arabinose.

FIG. 5 is a chromatogram of a derivative product of a sucrose sample as an auxiliary material, wherein peak 2 in FIG. 5 is rhamnose, peak 3(S) is glucose, peak 4 is galactose, and peak 5 is arabinose.

FIG. 6 is a second sample feature map.

FIG. 7 is a chromatogram of a mixed control.

FIG. 8 is a relative retention time calculation map.

FIG. 9 is a chromatogram of a mixed control in a specificity experiment.

FIG. 10 is a chromatogram of a sample in a specific experiment.

FIG. 11 is a chromatogram of a solid particle-sucrose negative control.

FIG. 12 is a chromatogram of a solid particle (batch No. Z200401).

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

In the following examples, thin layer chromatography identification:

1. apparatus and materials

Thin layer plates (10cm multiplied by 10cm), silica gel G, Qingdao sea (batch number: 20180305);

ginsenoside Rg1 reference (batch number: 330F021, HPLC ≥ 98%), Beijing Soilebao Tech Co., Ltd;

ginsenoside Rb1 reference substance (batch No. 705I025, HPLC ≥ 98%), Beijing Soilebao Tech Co., Ltd;

ginsenoside Re reference (batch No. 1118D022, HPLC ≥ 98%), Beijing Solaibao Tech Co., Ltd;

ginseng reed rhizome reference medicinal material (batch number: 121134-201702), China institute for testing and testing food and drug.

2. Reagent

Trichloromethane (AR, batch No. 20160316);

n-butanol (AR, batch No. 20180801);

methanol (AR, batch No. 20190111);

ethyl acetate (AR, lot number 20181109);

aqueous ammonia (AR, lot number: 20181019);

all purchased from Beijing chemical plant.

Water (whatpu ultrapure water machine).

Examples 1,

First, thin layer identification

Collecting powder (calculated by 1g of rhizoma Ginseng), adding chloroform 100ml, heating and refluxing for 1 hr, discarding chloroform solution, volatilizing solvent from residue, adding water 5ml, stirring, moistening, adding water saturated n-butanol 100ml, ultrasonic treating for 30 min, collecting supernatant, adding 2 times of ammonia solution, shaking, standing for layering, collecting supernatant, evaporating to dryness, and dissolving residue with methanol 5ml to obtain sample solution. And adding methanol into ginsenoside Rb1, ginsenoside Re, and ginsenoside Rg1 to obtain mixed solutions each containing 2mg of ginsenoside Rb1, ginsenoside Re, and ginsenoside Rg1 per 1 ml. Performing thin layer chromatography (0502) test, collecting 5 μ l, 5 μ l, and 5 μ l of the above three solutions, respectively dropping on the same silica gel G thin layer plate, spreading with lower layer solution of chloroform-ethyl acetate-water (15:40:22:10) at temperature below 10 deg.C as developing agent, taking out, air drying, spraying with 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, and respectively inspecting under sunlight and ultraviolet lamp (365 nm). In the chromatogram of the test solution, spots and fluorescent spots of the same color are respectively displayed at the positions corresponding to the chromatograms of the reference drug and the reference solution. The results are shown in FIGS. 1-2.

Second, characteristic atlas

1. Apparatus and materials

High performance liquid chromatography system: shimadzu LC-20 AD;

ginsenoside Rg1 reference (batch number: 330F021, HPLC ≥ 98%), Beijing Soilebao Tech Co., Ltd;

ginsenoside Rb1 reference substance (batch No. 705I025, HPLC ≥ 98%), Beijing Soilebao Tech Co., Ltd;

ginsenoside Re reference (batch No. 1118D022, HPLC ≥ 98%), Beijing Solaibao Tech Co., Ltd;

ginseng reed rhizome reference medicinal material (batch number: 121134-201702), China institute for testing and testing food and drug.

2. Reagent

Acetonitrile: fisher chemical ltd, usa, lot number: 197812, respectively;

methanol: fisher chemical ltd, usa, lot number: 195912, respectively;

chloroform (AR, batch No. 20160316), Beijing chemical plant;

n-butanol (AR, batch No. 20180801), Beijing chemical plant;

ammonia (AR, batch No. 20181019), Beijing chemical plant;

water: preparing the ultra-pure water machine of Waterppu.

3. Method for preparing test solution

Octadecylsilane chemically bonded silica is used as a filler in chromatographic conditions and system adaptability tests; acetonitrile is used as a mobile phase A, and 0.1% phosphoric acid is used as a mobile phase B. Specifically, gradient elution was carried out as specified in Table 1 below; flow rate: 1.0 mL/min; detection wavelength: 203 nm. The number of theoretical plates should not be less than 6000 calculated according to the peak of ginsenoside Re, and should not be less than 200000 calculated according to the peak of ginsenoside Rd.

TABLE 1 gradient elution

Preparation of reference solution ginsenoside Rg1 reference, ginsenoside Re reference, ginsenoside Rb1 reference, ginsenoside Rb2 reference, ginsenoside Rb3 reference, ginsenoside Rc reference, and ginsenoside Rd reference are precisely weighed, and methanol is added to obtain mixed solution containing 0.2mg of each 1 ml.

Preparing a sample solution, taking about 1g of the powder (passing through a sieve of No. four), precisely weighing, adding 50ml of methanol, carrying out ultrasonic treatment for 30 minutes (power is 250W and frequency is kHz), filtering, precisely taking 25ml of subsequent filtrate, evaporating to dryness, dissolving residues in 25ml of water-saturated n-butyl alcohol, placing in a separating funnel, adding 3 times of ammonia test solution for washing, shaking uniformly, standing for layering, taking n-butyl alcohol for evaporating to dryness, adding a methanol solution into residues, transferring to a 5ml measuring flask, adding methanol for diluting to a scale, shaking uniformly, filtering, and taking subsequent filtrate.

The determination method comprises precisely sucking 5 μ l of reference solution and sample solution respectively, injecting into liquid chromatograph, determining, collecting for 110min, and recording chromatogram.

Wherein, fig. 3 is a chromatogram of a derivative product of a test sample, and fig. 4 and 5 are a chromatogram of a derivative product of a mixed reference peak and a chromatogram of a derivative product of a sucrose sample as an auxiliary material, respectively, to serve as a reference of an absorption peak of a known compound therein.

As shown in FIG. 6, the sample characteristic spectrum should exhibit 7 characteristic peaks, and each characteristic peak should have the same retention time as the corresponding control peak (shown in FIG. 7).

The retention times of 7 characteristic peaks were compared as shown in Table 2 below.

TABLE 27 comparison of characteristic Peak Retention time

Relative retention time calculation, as shown in fig. 8.

And (4) conclusion: and (3) calculating the relative retention time of characteristic peaks 3-7 of the ginsenoside Rd reference peak, wherein the relative retention time is within +/-5% of a specified value: 0.84 (peak 3), 0.91 (peak 4), 0.93 (peak 5), 0.95 (peak 6), and 1.00 (peak 7), which are specifically shown in table 3 below.

TABLE 3

The above experimental methodology verifies that:

(1) repeatability of

According to the established extraction method of ginsenoside, the same test sample is prepared and tested for 6 times, and 6 test results are used for evaluation. The liquid chromatograms were separately injected and the peak areas (as shown in table 4) were recorded and the mean relative standard deviation was calculated.

TABLE 4

Calculated from the data in table 4, the average RSD (%) was 1.05.

The repeatability result RSD is less than 3 percent, and the technical requirements formulated by the traditional Chinese medicine quality standard research in Chinese pharmacopoeia are met.

(2) Intermediate precision

Precision degree

The same sample solution was sampled 6 times by 10. mu.l each, and the peak area was recorded (as shown in Table 5) to calculate the average relative standard deviation.

TABLE 5

Calculated from the data in table 5, the average RSD (%) was 1.09.

The above results show that the instrument used in the method is accurate.

(3) Specificity

The results are shown in FIGS. 9-10.

From the results shown in FIGS. 9 to 10, it can be seen that the retention times of the corresponding peaks of the chromatograms of the test solution and the control solution were consistent (the retention times of the specific peaks are shown in Table 6), the negative control was free from interference, and the separation degree between the adjacent components was good.

TABLE 6

(4) Stability test

And (5) carrying out stability investigation on the test sample. The test solutions were compared for peak area stability (specific peak areas are shown in table 7) of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc, ginsenoside Rb2, ginsenoside Rb3, and ginsenoside Rd after standing at room temperature for 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24 hours. The mean relative standard deviation was calculated.

TABLE 7

Calculated from the data in table 7, the average RSD (%) was 1.04.

The above results show that the sample is stable within 24 hours at room temperature.

III, molecular weight and molecular weight distribution

Sepax SRT SEC-100(8.0 mm. times.300 mm) gel column (Seidecto, USA) for chromatographic condition and system adaptability test; 0.7% sodium sulfate as mobile phase; the column temperature is 35 ℃, and the flow rate is 0.5 mL/min; the number of theoretical plates calculated by glucose peak should be not less than 3000, and the standard samples are Dextran-1000, Dextran-5000, Dextran-12000, Dextran-25000, Dextran-50000 and anhydrous glucose with known molecular weight.

Preparation of control solution A control solution was prepared by adding a mobile phase to a solution containing 5mg of Dextran of known molecular weight selected from Dextran-1000, Dextran-5000, Dextran-12000, Dextran-25000, Dextran-50000 and glucose anhydrous in an appropriate amount to 1 ml.

Preparation of test solution 10mg of the product was dissolved in 2mL of mobile phase and filtered through a 0.45 μm microporous membrane to obtain a test solution.

The measuring method precisely absorbs 20 mul of each solution of the test sample and the standard sample, and injects the solution into a chromatograph, records the chromatogram, and adopts special software of GPC polysaccharide to process.

The weight average molecular weight of the solid particles should be not less than 8000.

Precision:

a sample solution of Astragalus polysaccharides (lot number Z200301) was continuously injected 6 times to determine the molecular weight, as shown in Table 8.

TABLE 86 results of determination of molecular weight

As is clear from the results in Table 8, the sample solutions of the present invention were satisfactory in precision.

Stability:

a sample solution of Astragalus polysaccharides (lot No. Z200301) was sampled every 30 minutes to determine the molecular weight, and the results are shown in Table 9.

TABLE 9

From the results in Table 9, it is clear that the sample solution of the present invention has stability.

Repeatability:

6 parts of an Astragalus polysaccharide (lot No. Z200301) was precisely weighed, and the molecular weight was measured under the above-mentioned chromatographic conditions, and the results are shown in Table 10.

Watch 10

As is clear from the results in Table 10, the sample solutions of the present invention have reproducibility.

Specificity test:

taking 5mg of negative control sample lacking astragalus polysaccharide, preparing the negative control solution lacking astragalus polysaccharide according to the method for preparing the test solution drawn up according to the text. And (5) sample injection measurement, wherein a chromatographic peak appears at a position corresponding to the characteristic peak of the astragalus polysaccharide, which indicates that negative control is not interfered.

Ten batches of astragalus polysaccharide molecular weight determination

The results of measurement of astragalus polysaccharides according to the above chromatographic conditions are shown in table 11.

TABLE 11

As can be seen from the results of the above Table 11, the weight average molecular weight distribution of the ten batches of Astragalus polysaccharides was 8200-9300. The method for measuring the molecular weight and the molecular weight distribution of the astragalus polysaccharide is accurate, feasible, simple, convenient and quick, has high precision and good sample reproducibility, and can be used as a detection method for controlling the quality of the product. Therefore, it is set as a mass standard and is tentatively set to a weight average molecular weight of not less than 8000.

Claims (4)

1. A detection method for controlling the quality of vitality securing granules comprises the following steps:

(1) and (3) thin-layer chromatography detection: 1) extracting sample solid powder to obtain sample methanol solution, and respectively preparing reference methanol solutions of ginsenoside Rb1, ginsenoside Re, and ginsenoside Rg 1;

2) respectively measuring the methanol solution of the test sample and the methanol solution of the reference sample by adopting a thin-layer chromatography, and respectively displaying spots and fluorescent spots with the same color on corresponding positions of the two, so that the solid particles of the test sample are qualified; otherwise, the sample solid particles are unqualified;

(2) and (3) characteristic map identification: 1) treating the sample solid particle powder to prepare a sample solution a; preparing reference solution a from the ginsenoside Rg1 reference, the ginsenoside Re reference, the ginsenoside Rb1 reference, the ginsenoside Rb2 reference, the ginsenoside Rb3 reference, the ginsenoside Rc reference and the ginsenoside Rd reference respectively;

2) respectively detecting the test solution a and the reference solution a by liquid chromatography, wherein 7 characteristic peaks are presented in a map of the test solution a, and the retention time of each characteristic peak is the same as that of the reference solution a, so that the test solid particles are qualified; otherwise, the sample solid particles are unqualified;

the conditions of the liquid chromatography were as follows: octadecylsilane chemically bonded silica is used as a filling agent;

acetonitrile is taken as a mobile phase A, and 0.1% phosphoric acid is taken as a mobile phase B;

flow rate: 1.0 mL/min; detection wavelength: 203 nm;

gradient elution was performed as follows in volume percent: 0min, mobile phase A18%, mobile phase B82%; 40min, mobile phase A18%, mobile phase B82%; 50min, mobile phase A22%, mobile phase B78%; 70min, mobile phase A28%, mobile phase B72%; 100min, mobile phase A38%, mobile phase B62%; 110min, mobile phase A18%, mobile phase B82%;

(3) determination of molecular weight and molecular weight distribution: 1) adding mobile phase into dextran-1000, dextran-5000, dextran-12000, dextran-25000, dextran-50000 and anhydrous glucose with known molecular weight to obtain solutions each containing 5mg per 1ml as reference solution b;

taking 10mg of the sample solid particle powder which is qualified through the steps (1) and (2) in sequence or the steps (2) and (1) in sequence, adding 2 mL0.7% of sodium sulfate for dissolution, and filtering by using a 0.45-micron microporous filter membrane to obtain a sample solution b;

2) the reference substance solution b and the test substance solution b are measured by chromatography, chromatogram is recorded, and GPC polysaccharide special software is adopted for processing; when the weight average molecular weight of the sample solid particles is 3000-8000, the sample solid particles are qualified; otherwise, the sample solid particles are unqualified;

the chromatographic conditions were as follows: sepax SRT SEC-100(8.0mm × 300mm) gel column; 0.7% sodium sulfate as mobile phase; the column temperature is 35 ℃, and the flow rate is 0.5 mL/min; differential refractive index detector, LZD GPC2010 dedicated software workstation, theoretical plate number calculated as glucose peak.

2. The method of claim 1, wherein: in the step (1) -1), the extraction treatment of the sample solid particles is as follows: taking the sample solid particle powder, counting according to 1g of ginseng head, adding 100ml of trichloromethane, heating and refluxing for 1 hour, removing chloroform liquid, volatilizing solvent from decoction dregs, adding 5ml of water, uniformly stirring and wetting, adding 100ml of water-saturated n-butyl alcohol, carrying out ultrasonic treatment for 30 minutes, absorbing supernatant, adding 2 times of ammonia sample, shaking uniformly, standing for layering, taking supernatant, evaporating to dryness, and adding 5ml of methanol to dissolve residues to obtain a sample solution;

the reference methanol solution is prepared by mixing ginsenoside Rb1 reference, ginsenoside Re reference, and ginsenoside Rg1 reference with methanol to obtain mixed solution containing 2mg of each 1 ml.

3. The method according to claim 1 or 2, characterized in that: the thin layer chromatography: sucking three kinds of control methanol solutions 5 μ l each, respectively dropping on the same silica gel G thin layer plate, developing with chloroform-ethyl acetate-water (15:40:22:10) lower layer solution at 10 deg.C below as developing agent, taking out, air drying, spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, and respectively inspecting under sunlight and ultraviolet lamp (365 nm).

4. The method according to any one of claims 1-3, wherein: (2) the method for treating the test article solid particle powder in the 1) comprises the following steps:

taking 1g of the test sample solid particle powder passing through a No. four sieve, adding 50ml of methanol, and carrying out ultrasonic treatment for 30 minutes at the power of 250W and the frequency of kHz; filtering, measuring 25ml of subsequent filtrate, evaporating to dryness, dissolving the residue in 25ml of water-saturated n-butanol, placing in a separating funnel, adding 3 times of ammonia test solution, washing, shaking uniformly, standing for layering, taking n-butanol, evaporating to dryness, adding methanol solution into the residue, transferring to a 5ml measuring flask, adding methanol to dilute to a scale, shaking uniformly, filtering, and taking subsequent filtrate to obtain the sample solution a.

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