A quality detection method for red ginseng Chinese medicinal slices

Abstract

The invention discloses a quality detection method for red ginseng Chinese medicinal slices, the method comprising: preparing a test solution, preparing a reference solution, and calculating the contents of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin by an external standard method. The quality detection method for red ginseng Chinese medicinal slices provided by the invention screens out the best mobile phase composition, gradient elution program, flow rate, detection wavelength, chromatographic column, column temperature and other analysis conditions through a large number of experiments. It is shown through multiple experimental verifications that the detection method provided by the invention has high accuracy, good stability and good repeatability, and can simultaneously detect the contents of three ginsenoside components and schisandrin A. The quality of red ginseng Chinese medicinal slices can be evaluated more objectively, comprehensively and accurately, thereby more effectively ensuring the quality of red ginseng Chinese medicinal slices.

Description

Technical Field

[0001] The invention relates to a quality detection method for Chinese medicinal materials, and in particular to a quality detection method for red ginseng Chinese medicinal slices. Background Art

[0002] Red ginseng is a traditional Chinese medicine with multiple functions such as restoring pulse and strengthening deficiency, replenishing vital energy, replenishing qi and blood, strengthening the heart, promoting body fluid, strengthening the stomach, and calming the mind. The active ingredients contained in red ginseng are mainly ginsenosides and polysaccharides. The detection of red ginseng in the Chinese Pharmacopoeia is the detection of a single ginsenoside. Most of the existing technical papers are also the detection of a single ginsenoside compound.

[0003] The present invention uses high performance liquid chromatography to establish a method for simultaneously determining the contents of three ginsenoside components and schisandrin A in red ginseng Chinese medicinal slices, thereby providing a basis for better monitoring the quality and clinical effectiveness of red ginseng Chinese medicinal slices. Summary of the invention

[0004] Purpose of the invention: In order to solve the shortcomings of the prior art, the present invention provides a detection method with high precision, good repeatability and stability, which can objectively, comprehensively and accurately evaluate the quality of red ginseng Chinese herbal medicine slices, thereby more effectively ensuring the quality of red ginseng Chinese herbal medicine slices.

[0005] Technical solution: In order to achieve the above purpose, the technical solution adopted by the present invention is:

[0006] A method for quality testing of red ginseng Chinese medicinal slices, characterized in that the method comprises the following steps:

[0007] (1) Preparation of mixed reference solution

[0008] Accurately weigh ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin A reference substances, and add methanol to prepare mixed reference substance solutions containing 0.10-0.30 mg, 0.08-0.24 mg, 0.20-0.60 mg and 0.02-0.06 mg per ml respectively;

[0009] (2) Preparation of test solution

[0010] Take the red ginseng Chinese medicine slices, crush them appropriately, add water or ethanol to extract, combine the extracts, pass through a microporous filter membrane, and use them as the test solution;

[0011] (3) Determination of test solution

[0012] Accurately pipette 10 μl of the reference solution of step (1) and the test solution of step (2), respectively, and inject them into the high performance liquid chromatograph, and calculate the contents of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin by the external standard method.

[0013] As a preferred embodiment, in the quality detection method of the red ginseng Chinese medicinal slices described above, the preparation method of the mixed reference solution in step (1) is:

[0014] Accurately weigh ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin A reference substances, and add methanol to prepare mixed reference substance solutions containing 0.10 mg, 0.08 mg, 0.20 mg and 0.02 mg per ml, respectively.

[0015] As a preferred embodiment, in the above-mentioned method for quality detection of red ginseng Chinese medicinal slices, the method for preparing the test solution in step (2) is:

[0016] Take red ginseng Chinese medicinal slices, crush them appropriately, add 6 to 15 times the weight of the medicinal material water or ethanol to extract 1 to 3 times, each time for 0.5 to 2 hours, combine the extracts, pass through a microporous filter membrane, and use them as the test solution.

[0017] As a preferred embodiment, in the quality inspection method for the red ginseng Chinese medicinal slices described above, the extraction method in step (2) is a decoction method when water is used as the solvent; when ethanol is used as the extraction solvent, the extraction method is a reflux extraction method, a continuous reflux extraction method or an ultrasonic extraction method.

[0018] As a preferred embodiment, in the quality detection method of the red ginseng Chinese medicinal slices described above, the chromatographic conditions in step (3) are:

[0019] The stationary phase uses Waters symmetry shield TM RP 18 Chromatographic column (4.6 mm × 250 mm; 5.0 μm); column temperature 30 ° C, acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the table below, detection wavelength 203 nm;

[0020]

[0021] As a preferred embodiment, in the quality detection method of the red ginseng Chinese medicinal slices described above, the calculation formula of the external standard method in step (3) is:

[0022]

[0023] in: Average correction factor for reference substances

[0024] f R : Reference correction factor

[0025] C R :Concentration of reference solution (mg / ml)

[0026] A X: Peak area of ​​test sample

[0027] A R : Peak area of ​​reference substance.

[0028] Screening of chromatographic conditions:

[0029] 1. Wavelength selection

[0030] The present invention firstly performs full wavelength scanning with DAD and selects the maximum absorption wavelength of 203nm of three ginsenosides and one schisandrin A, at which the four components can be determined simultaneously without interference.

[0031] 2. Selection of mobile phase

[0032] The present invention investigates a variety of mobile phase systems such as methanol-water, methanol-0.05% phosphoric acid water, acetonitrile-0.05% water, methanol-acetonitrile, etc. The results show that when acetonitrile-water solution is used as the mobile phase, the separation effect of the components in the red ginseng Chinese medicinal slices is better, so acetonitrile and water are finally selected as the mobile phase.

[0033] After determining the optimal flowability composition, the present invention continues to screen different gradient elution methods, and through experiments it is found that the best separation degree is obtained when 0 to 30 min, 0 to 10% acetonitrile; 30 to 40 min, 10 to 23% acetonitrile; 40 to 50 min, 23% acetonitrile; 50 to 85 min, 23 to 60% acetonitrile; 85 to 95 min, 60 to 100% acetonitrile is used.

[0034] 3. Selection of column temperature and flow rate

[0035] The present invention investigates the influence of different temperatures (25°C, 30°C, 35°C and 40°C) and different volume flow rates (0.6, 0.8, 1.0 and 1.2 mL / min) on the separation effect of each liquid chromatographic peak. The results show that when the column temperature is 30°C and the volume flow rate is 1.0 mL / min, the baseline can be stable and the separation degree and peak shape are better.

[0036] Beneficial effects: The quality detection method of red ginseng Chinese medicinal slices provided by the present invention has the following advantages over the prior art:

[0037] The quality detection method for red ginseng Chinese medicinal slices provided by the present invention screens out the best mobile phase composition, gradient elution program, flow rate, detection wavelength, chromatographic column, column temperature and other analysis conditions through a large number of experiments. Multiple experimental verifications show that the detection method provided by the present invention has high accuracy, good stability and good repeatability; and can simultaneously detect the content of three ginsenoside components and schisandrin A. DETAILED DESCRIPTION

[0038] The present invention is further explained below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. After reading the present invention, various equivalent modifications of the present invention by those skilled in the art all fall within the scope defined by the claims attached to this application.

[0039] Example 1

[0040] A method for quality testing of red ginseng Chinese medicinal slices, characterized in that the method comprises the following steps:

[0041] (1) Preparation of mixed reference solution

[0042] Accurately weigh ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin A reference substances, respectively, and add methanol to prepare mixed reference substance solutions containing 0.10 mg, 0.08 mg, 0.20 mg and 0.02 mg per ml respectively;

[0043] (2) Preparation of test solution

[0044] Take the red ginseng slices, crush them appropriately, add 12 times the weight of the herbs in water and boil them for extraction twice, each time for 1 hour, combine the extracts, filter through a microporous membrane, and use them as the test solution;

[0045] (3) Determination of test solution

[0046] Accurately pipette 10 μl of the reference solution in step (1) and the test solution in step (2), inject them into the high performance liquid chromatograph, and calculate by the external standard method. The calculation formula of the external standard method is:

[0047]

[0048] in: Average correction factor for reference substances

[0049] f R : Reference correction factor

[0050] C R :Concentration of reference solution (mg / ml)

[0051] A X : Peak area of ​​test sample

[0052] A R : Peak area of ​​reference substance.

[0053] Contains ginsenoside Rg1(C 42 H 72 O 14 ) content is 0.23mg / g, ginsenoside Re(C 48 H 82 O18 ) content is 0.4mg / g, ginsenoside Rb1(C 54 H 92 O 23 )

[0054] The content is 0.36mg / g,

[0055] The chromatographic conditions of step (3) above are:

[0056] The stationary phase uses Waters symmetry shield TM RP 18 Chromatographic column (4.6 mm × 250 mm; 5.0 μm); column temperature 30 ° C, acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the table below, detection wavelength 203 nm;

[0057]

Claims

1. A quality detection method for red ginseng Chinese medicinal slices, characterized in that: The method comprises the following steps: (1) Preparation of mixed reference solution Accurately weigh ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin A reference substances, and add methanol to prepare mixed reference substance solutions containing 0.10-0.30 mg, 0.08-0.24 mg, 0.20-0.60 mg and 0.02-0.06 mg per ml respectively; (2) Preparation of test solution Take the red ginseng Chinese medicine slices, crush them appropriately, add water or ethanol to extract, combine the extracts, pass through a microporous filter membrane, and use them as the test solution; (3) Determination of test solution Accurately pipette 10 μl of the reference solution of step (1) and the test solution of step (2), respectively, and inject them into the high performance liquid chromatograph, and calculate the contents of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin by the external standard method; The chromatographic conditions of the HPLC were: The stationary phase uses Waterssymmetryshield TM RP 18 Chromatographic column, the specification of the chromatographic column is 4.6mm×250mm; 5.0μm; the column temperature is 30℃, acetonitrile is used as mobile phase A, water is used as mobile phase B, gradient elution is performed as shown in the table below, and the detection wavelength is 203nm; 2. The quality detection method of red ginseng Chinese medicinal slices according to claim 1, characterized in that: The preparation method of the mixed reference substance solution in step (1) is: Accurately weigh ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and schisandrin A reference substances, and add methanol to prepare mixed reference substance solutions containing 0.10 mg, 0.08 mg, 0.20 mg and 0.02 mg per ml, respectively.

3. The quality detection method of red ginseng Chinese medicinal slices according to claim 2, characterized in that: The preparation method of the test solution in step (2) is: Take red ginseng Chinese medicinal slices, crush them appropriately, add 6 to 15 times the weight of the medicinal material water or ethanol to extract 1 to 3 times, each time for 0.5 to 2 hours, combine the extracts, pass through a microporous filter membrane, and use them as the test solution.

4. The quality detection method of red ginseng Chinese medicinal slices according to claim 1, characterized in that: When water is used as the extraction solvent in step (2), the extraction method is a decoction method; when ethanol is used as the extraction solvent, the extraction method is a reflux extraction method, a continuous reflux extraction method or an ultrasonic extraction method.

5. The quality detection method of red ginseng Chinese medicinal slices according to claim 1, characterized in that: The calculation formula of step (3) using the external standard method is: in: Average correction factor for reference substances f R : Reference correction factor C R : Concentration of reference solution mg / ml A X : Peak area of ​​test sample A R : Peak area of ​​reference substance.

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