A multi-component quality control method for Tianfoshen oral liquid

Abstract

The present invention discloses a multi-component quality control method for Tianfoshen oral liquid, which comprises (1) preparing a test solution: (2) preparing a mixed reference solution; determining the HPLC fingerprint of the Tianfoshen oral liquid and calibrating the effective components. The present invention screens out the best mobile phase composition, detection wavelength, chromatographic column, column temperature and other analytical conditions through a large number of experiments. It is shown through multiple experimental verifications that the detection method provided by the present invention can simultaneously detect multiple active components such as ginsenosides and asparagus saponins with similar polarity, so that the quality of the Tianfoshen oral liquid can be comprehensively, objectively and accurately detected and evaluated, which is of great significance for ensuring clinical efficacy.

Description

Technical Field

[0001] The invention relates to a quality detection method for a traditional Chinese medicine preparation, and in particular to a multi-component quality control method for a Tianfoshen oral liquid. Background Art

[0002] Tianfoshen oral liquid is a pure natural Chinese medicine preparation extracted and refined by modern advanced technology. It has the effects of nourishing yin and qi, detoxifying and dispersing knots. When used in combination with anti-tumor chemicals, it can alleviate nausea, vomiting, constipation, hair loss and other phenomena caused by chemotherapy. Tianfoshen oral liquid is composed of 8 medicinal materials: American ginseng, toad venom, asparagus, Fritillaria thunbergii, Acanthopanax obovata, kiwi root, sea buckthorn fruit, and bergamot. It contains saponins, flavonoids, alkaloids and other effective ingredients. The existing content detection methods can detect few types of effective ingredients. It is not possible to comprehensively and objectively detect and evaluate the quality of Tianfoshen oral liquid. Summary of the invention

[0003] Purpose of the invention: The purpose of the present invention is to solve the deficiencies of the prior art and to provide a multi-component quality control method for Tianfoshen oral liquid. This detection method can objectively, comprehensively and accurately evaluate the quality of Tianfoshen oral liquid, which is of great significance for controlling the intrinsic quality of Tianfoshen oral liquid and ensuring clinical efficacy.

[0004] Technical solution: In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0005] A method for controlling the quality of multiple components of Tianfoshen oral liquid, characterized in that it comprises the following steps:

[0006] (1) Preparation of test solution: Take Tianfoshen oral solution and add methanol to dissolve it by ultrasonic, filter it through a 0.22 μm microporous membrane, and take the filtrate to prepare the test solution;

[0007] (2) Preparation of mixed reference solution

[0008] Take ginsenoside Rb1, Rb2, Rc, Rd, Rh2, Re, Rg1, asparagine-Ⅳ, asparagine-Ⅳ, ginsenoside A, ginsenoside B respectively, dilute to a certain concentration with methanol, filter through a 0.22 μm microporous membrane, and set aside;

[0009] (3) Determination of HPLC fingerprint of Tianfoshen oral liquid: Accurately aspirate the test solution of Shengmai injection in step (1) and the mixed reference solution in step (2), inject them into the high performance liquid chromatograph, and record the chromatogram;

[0010] (4) The fingerprint of the Tianfoshen oral liquid obtained in step (3) is exported and imported into the Chinese medicine chromatographic fingerprint similarity evaluation system 2012A; the chromatographic peaks existing in the chromatograms of different batches of Tianfoshen oral liquid are selected as common peaks; the reference fingerprint of the Tianfoshen oral liquid is generated by the average value calculation method, and the relative retention time and relative peak area of ​​each common peak are calculated; and the chemical composition of the peak in the reference fingerprint is marked according to the retention time of the chromatogram of the mixed reference solution.

[0011] As a preferred embodiment, the above-mentioned Tianfoshen oral liquid multi-component quality control method, the test solution is prepared: take Tianfoshen oral liquid and add 8 to 16 times the amount of methanol to dissolve it by ultrasonic for 10 to 30 minutes, take the filtrate and filter it through a 0.22μm microporous filter membrane, take the subsequent filtrate, and prepare the test solution.

[0012] As a preferred embodiment, the method for preparing the mixed reference solution of the multi-component quality control method of Tianfoshen oral liquid described above is as follows:

[0013] Accurately weigh appropriate amounts of ginsenoside Rb1, Rb2, Rc, Rd, Rh2, Re, Rg1, asparagine-Ⅳ, asparagine-Ⅳ, radix fritillariae saponin A, and radix fritillariae saponin B, add methanol to dilute them into mixed reference solutions with concentrations of 1.24 mg / ml, 1.89 mg / ml, 2.04 mg / ml, 1.79 mg / ml, 1.56 mg / ml, 3.51 mg / ml, 2.78 mg / ml, 2.49 mg / ml, 4.21 mg / ml, 1.59 mg / ml, and 2.25 mg / ml, respectively, filter through a 0.22 μm microporous filter membrane and set aside.

[0014] As a preferred embodiment, in the above-mentioned Tianfoshen oral liquid multi-component quality control method, in step (3), the chromatographic conditions are: chromatographic column: 11STAR LP RP-18 endcapped, specification is 4.6*250mm, 5μm, methanol is used as mobile phase A, and 0.05% volume concentration of phosphoric acid aqueous solution is used as mobile phase B, gradient elution; column temperature is 30°C; flow rate is 1ml per minute; detection wavelength: 265nm.

[0015] As a preferred embodiment, in the above-mentioned method for controlling the quality of multi-components of Tianfoshen oral liquid, the gradient elution procedure in step (3) is:

[0016] Time (minutes) Mobile phase A(﹪) Mobile phase B (﹪) 0～8.0 10 90 8.0～15 13 87 15.0～30.0 20 80 30.0～52.0 63 37 52.0～68 42 58 68～90 33 77 .

[0017] Beneficial effects: The multi-component quality control method of Tianfoshen oral liquid provided by the present invention has the following advantages:

[0018] The present invention aims at the many deficiencies of the detection methods in the prior art, and screens out the optimal mobile phase composition, detection wavelength, chromatographic column, column temperature and other analytical conditions through a large number of experiments. It is shown through multiple experimental verifications that the detection method provided by the present invention can simultaneously detect multiple active ingredients such as ginsenosides and asparagus saponins with similar polarity, thereby being able to comprehensively, objectively and accurately detect and evaluate the quality of Tianfoshen oral liquid, which is of great significance for ensuring clinical efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is the reference fingerprint spectrum I of the anti-tumor Chinese medicine composition of the present invention. DETAILED DESCRIPTION

[0020] 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.

[0021] Example 1

[0022] A method for controlling the quality of multiple components of Tianfoshen oral liquid, characterized in that it comprises the following steps:

[0023] (1) Preparation of test solution: Take Tianfoshen oral solution and add 15 times the amount of methanol to dissolve it by ultrasonic for 30 minutes, take the filtrate and filter it through a 0.22 μm microporous membrane, take the filtrate and prepare the test solution;

[0024] (2) Preparation of mixed reference solution

[0025] Accurately weigh appropriate amounts of ginsenoside Rb1, Rb2, Rc, Rd, Rh2, Re, Rg1, asparagine-Ⅳ, asparagine-Ⅳ, radix radix saponin A, and radix radix saponin B, respectively, add methanol to dilute to mixed reference solutions with concentrations of 1.24 mg / ml, 1.89 mg / ml, 2.04 mg / ml, 1.79 mg / ml, 1.56 mg / ml, 3.51 mg / ml, 2.78 mg / ml, 2.49 mg / ml, 4.21 mg / ml, 1.59 mg / ml, and 2.25 mg / ml, respectively, filter through a 0.22 μm microporous filter membrane, and set aside;

[0026] (3) Determination of HPLC fingerprint of Tianfoshen oral liquid: Accurately aspirate the test solution of Shengmai injection in step (1) and the mixed reference solution in step (2), inject them into the high performance liquid chromatograph, and record the chromatogram;

[0027] The chromatographic conditions are as follows: chromatographic column: 11STAR LP RP-18 endcapped, specification: 4.6*250mm, 5μm, methanol as mobile phase A, 0.05% volume concentration of phosphoric acid aqueous solution as mobile phase B, gradient elution; column temperature: 30°C; flow rate: 1ml per minute; detection wavelength: 265nm; the gradient elution program in step (3) is:

[0028] Time (minutes) Mobile phase A(﹪) Mobile phase B (﹪) 0～8.0 10 90 8.0～15 13 87 15.0～30.0 20 80 30.0～52.0 63 37 52.0～68 42 58 68～90 33 77

[0029] (4) The fingerprint of Tianfoshen oral liquid obtained in step (3) is exported and imported into the Chinese medicine chromatographic fingerprint similarity evaluation system 2012A; the chromatographic peaks existing in the chromatograms of different batches of Tianfoshen oral liquid are selected as common peaks; the reference fingerprint of Tianfoshen oral liquid is generated by the average value calculation method, a total of 35 common chromatographic peaks are identified, and the relative retention time and relative peak area of ​​each common peak are calculated; and the chemical components of the peaks in the reference fingerprint are marked according to the retention time of the chromatogram of the mixed reference solution. Among them, ginsenoside Rd is peak 3, ginsenoside Rh2 is peak 4, ginsenoside Re is peak 6, ginsenoside Rg1 is peak 11, ginsenoside Rb1 is peak 16, ginsenoside Rb2 is peak 21, ginsenoside Rc is peak 23, asparagine-IV is peak 20, asparagine-IV is peak 22, scutellariae saponin A is peak 14, and scutellariae saponin B is peak 18.

[0030] The above experimental results indicate that the multi-component quality control method for Tianfoshen oral liquid provided by the present invention can objectively, comprehensively and specifically analyze and evaluate the quality of Tianfoshen oral liquid, which is of great significance for ensuring clinical efficacy and has achieved significant technological progress.

[0031] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A multi-component quality control method for Tianfoshen oral liquid, characterized in that: The following steps are involved: (1) Preparation of test solution: Take Tianfoshen oral solution, add 8 to 16 times the amount of methanol to dissolve it by ultrasonic for 10 to 30 minutes, take the filtrate and filter it through a 0.22 μm microporous membrane, take the filtrate and prepare the test solution; (2) Preparation of mixed reference solution Take ginsenoside Rb1, Rb2, Rc, Rd, Rh2, Re, Rg1, asparagine-Ⅳ, asparagine-Ⅳ, ginsenoside A, ginsenoside B respectively, dilute to a certain concentration with methanol, filter through a 0.22 μm microporous membrane, and set aside; (3) Determination of HPLC fingerprint of Tianfoshen oral liquid: Accurately aspirate the test solution of Shengmai injection in step (1) and the mixed reference solution in step (2), inject them into the high performance liquid chromatograph, and record the chromatogram; The chromatographic conditions were as follows: chromatographic column: 11STAR LP RP-18 endcapped, specification: 4.6*250mm, 5μm, methanol as mobile phase A, 0.05% volume concentration of phosphoric acid aqueous solution as mobile phase B, gradient elution; column temperature: 30°C; flow rate: 1ml per minute; detection wavelength: 265nm; gradient elution program: (4) The fingerprint of the Tianfoshen oral liquid obtained in step (3) is exported and imported into the Chinese medicine chromatographic fingerprint similarity evaluation system 2012A; the chromatographic peaks existing in the chromatograms of different batches of Tianfoshen oral liquid are selected as common peaks; the reference fingerprint of the Tianfoshen oral liquid is generated by the average value calculation method, and the relative retention time and relative peak area of ​​each common peak are calculated; and the chemical composition of the peak in the reference fingerprint is marked according to the retention time of the chromatogram of the mixed reference solution.

2. The multi-component quality control method of Tianfoshen oral liquid according to claim 1, characterized in that: The preparation method of the mixed reference solution is as follows: Accurately weigh appropriate amounts of ginsenoside Rb1, Rb2, Rc, Rd, Rh2, Re, Rg1, asparagine-Ⅳ, asparagine-Ⅳ, radix fritillariae saponin A, and radix fritillariae saponin B, add methanol to dilute them into mixed reference solutions with concentrations of 1.24 mg / ml, 1.89 mg / ml, 2.04 mg / ml, 1.79 mg / ml, 1.56 mg / ml, 3.51 mg / ml, 2.78 mg / ml, 2.49 mg / ml, 4.21 mg / ml, 1.59 mg / ml, and 2.25 mg / ml, respectively, filter through a 0.22 μm microporous filter membrane and set aside.

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