Quality detection method of Juanbi decoction

Abstract

The invention provides a quality detection method of Juanbi decoction, and belongs to the technical field of medicine detection.The quality detection method comprises the steps that the Juanbi decoction is used for preparing a test solution, acetonitrile (A)-0.04-0.06 wt% formic acid aqueous solution (B) serves as a mobile phase, and the quality of the Juanbi decoction is detected through high performance liquid chromatography-tandem mass spectrometry. According to the invention, a liquid chromatography-mass spectrometry method is adopted to qualitatively analyze the chemical components of the Juanbi decoction material standard, so that a scientific basis is provided for the material basic research of the Juanbi decoction material standard, and meanwhile, the main chemical components of the Juanbi decoction are clarified.

Description

Technical Field

[0001] The present invention relates to drug detection technology, and in particular to a quality detection method for Juanbi Decoction. Background Art

[0002] The prescription of Juanbi Decoction is composed of Notopterygium incisum, Heracleum hemsleyanum, Gentiana macrophylla, Piper kadsura, Ligusticum wallichii, Cinnamomum cassia, Olibanum processed with vinegar, Aucklandia lappa, Angelica sinensis, Mori Ramulus, and Glycyrrhiza uralensis Fisch. (processed with honey). It is mainly used for treating stroke with general body pain, stiffness of the neck and back, cold numbness of hands and feet, heaviness of the waist and knees, and difficulty in movement. Juanbi Decoction is the 79th among the 100 classical famous prescriptions in the "Catalog of Classical Famous Prescriptions in Ancient China (the First Batch)". Classical famous prescriptions have definite clinical curative effects. Developing classical famous prescriptions into convenient-to-carry and easy-to-take traditional Chinese medicine preparations by using modern scientific and technological means is not only the inheritance of traditional Chinese medicine but also can better promote the clinical application of classical famous prescriptions.

[0003] High-performance liquid chromatography tandem mass spectrometry technology has high analysis efficiency and fast speed, can obtain accurate molecular structure information, and can obtain the first-order and multi-order mass spectrometry information of each compound, providing a good technical means for the multi-index component analysis of complex traditional Chinese medicine systems. Juanbi Decoction is made from 11 traditional Chinese medicines, and its components are extremely complex. At present, it is still impossible to comprehensively monitor the quality of Juanbi Decoction, let alone conduct qualitative analysis on the chemical components contained in Juanbi Decoction. Therefore, it is necessary to develop a quality detection method for Juanbi Decoction to more comprehensively evaluate the quality of Juanbi Decoction. Summary of the Invention

[0004] In view of the above problems, the present invention provides a quality detection method for Juanbi Decoction.

[0005] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0006] A quality detection method for Juanbi Decoction, comprising the following steps:

[0007] S1. Take Juanbi Decoction to prepare a test solution;

[0008] S2. Take the test solution and use high-performance liquid chromatography tandem mass spectrometry to detect the quality of Juanbi Decoction;

[0009] The mobile phase A for the high-performance liquid chromatography detection is acetonitrile, and the mobile phase B is an aqueous formic acid solution with a concentration of 0.04 - 0.06 wt%;

[0010] The elution method for the high-performance liquid chromatography detection is gradient elution;

[0011] The elution conditions for the gradient elution are as follows:

[0012] 0 - 12 min, 1% → 11% mobile phase A, 99% → 89% mobile phase B;

[0013] 12 - 14 min, mobile phase A: 11% → 13%, mobile phase B: 89% → 87%;

[0014] 14 - 15 min, mobile phase A: 13% → 14%, mobile phase B: 87% → 86%;

[0015] 15 - 18.5 min, mobile phase A: 14% → 16%, mobile phase B: 86% → 84%;

[0016] 18.5 - 21.5 min, mobile phase A: 16%, mobile phase B: 84%;

[0017] 21.5 - 25 min, mobile phase A: 16% → 19%, mobile phase B: 84% → 81%;

[0018] 25 - 26 min, mobile phase A: 19% → 26%, mobile phase B: 81% → 74%;

[0019] 26 - 32 min, mobile phase A: 26%, mobile phase B: 74%;

[0020] 32 - 33.5 min, mobile phase A: 26% → 44%, mobile phase B: 74% → 56%;

[0021] 33.5 - 45 min, mobile phase A: 44% → 46%, mobile phase B: 56% → 54%;

[0022] 45 - 46 min, mobile phase A: 46% → 66%, mobile phase B: 54% → 34%;

[0023] 46 - 55 min, mobile phase A: 66%, mobile phase B: 34%.

[0024] Furthermore, the detection wavelength for the high - performance liquid chromatography detection is 290 - 330 nm.

[0025] Furthermore, the flow rate for the high - performance liquid chromatography detection is 0.38 - 0.45 mL / min and the column temperature is 38 - 45 °C.

[0026] Furthermore, the chromatographic column for the high - performance liquid chromatography detection is packed with octadecylsilyl - bonded silica gel.

[0027] Furthermore, the mass spectrometry detection is carried out in positive ion mode and negative ion mode respectively.

[0028] Further, the conditions for mass spectrometry detection are as follows: the drying gas temperature is 345 - 355°C, the drying gas flow rate is 8 - 12 L / min, the nebulizing gas pressure is 34 - 36 psi, the sheath gas temperature is 345 - 355°C, the sheath gas flow rate is 11 - 13 L / min, the capillary voltage in positive ion mode is 3400 - 3600 V, and the capillary voltage in negative ion mode is 3400 - 3600 V.

[0029] Further, during the mass spectrometry detection, the first - stage mass spectrometry uses the MS mode with a mass scanning range of 100 - 1200 m / z, and the second - stage mass spectrometry uses the Auto MSMS mode.

[0030] Further, the test solution is prepared by taking solid Juanbi Decoction, adding methanol aqueous solution, performing ultrasonic treatment, and filtering.

[0031] The concentration of the methanol aqueous solution is 70 - 85 vol%.

[0032] The solid Juanbi Decoction is Juanbi Decoction dry extract powder, and the weight - to - volume ratio of Juanbi Decoction to the methanol aqueous solution is 0.9 - 1.1 g:10 mL.

[0033] Further, the test solution is prepared by taking Juanbi Decoction extract or Juanbi Decoction concentrated solution, adding methanol, performing ultrasonic treatment, and filtering.

[0034] Further, the volume ratio of Juanbi Decoction extract / Juanbi Decoction concentrated solution to methanol is 1:3.8 - 4.2.

[0035] The beneficial effects of the quality detection method of Juanbi Decoction of the present invention are as follows:

[0036] The present invention uses the liquid chromatography - mass spectrometry method to qualitatively analyze the chemical components of the material basis of Juanbi Decoction, providing a scientific basis for the study of the material basis of Juanbi Decoction and clarifying its main chemical composition at the same time.

[0037] The present invention uses the liquid chromatography - mass spectrometry method to qualitatively analyze the chemical components of the reference substance of Juanbi Decoction. It has high analysis efficiency and speed, can obtain accurate molecular structure information in Juanbi Decoction, and can obtain the first - stage and multi - stage mass spectrometry information of each compound, providing a good technical means for the multi - index component analysis of identifying the complex traditional Chinese medicine system of Juanbi Decoction.

[0038] The present invention uses the liquid chromatography - mass spectrometry method to qualitatively analyze the chemical components of the reference substance of Juanbi Decoction, providing a scientific basis for the study of the material basis of Juanbi Decoction and clarifying its main chemical composition at the same time.

[0039] The quality detection method of the present invention can jointly detect almost all the medicinal material components in the whole formula of Juanbi Decoction.

[0040] The present invention utilizes the characteristics of different physicochemical properties of the active ingredients in Juanbi Decoction, changes the chromatographic conditions, so as to achieve the purpose of separating different active ingredients in Juanbi Decoction well;

[0041] By selecting specific chromatographic conditions and mass spectrometry conditions, the present invention can obtain a better chromatogram of Juanbi Decoction;

[0042] The quality detection method of the present invention also has good feasibility, stability and reproducibility;

[0043] The quality detection method of the present invention can detect and clarify the origin attribution of the drug flavors of 41 peak substances and the qualitative analysis of the main chromatographic peaks; among them, 11 coumarin compounds were identified, namely decursin isomers, decursin, notopterol isomers, notopterol, which belong to the characteristic components of Notopterygium; iso-decursin, angelol G / D / B, angelol A, angelol C / F / E / H, anomertin, osthol, columbianadin, which belong to the characteristic components of Angelica pubescens; 5 iridoid glycoside compounds were identified, namely loganic acid, picroside I, gentiopicroside, 6'-O-β-D-glucopyranosyl gentiopicroside, picroside II, which belong to the characteristic components of Gentiana macrophylla; 5 phenylpropanoid compounds were identified, among which chlorogenic acid is a common component of Angelica sinensis, Angelica pubescens, Notopterygium incisum and Aucklandia lappa, neochlorogenic acid is a common component of Angelica pubescens and Aucklandia lappa, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, which belong to the exclusive components of Aucklandia lappa; 5 flavonoid compounds were identified, morusin is the exclusive component of Ramulus Mori, liquiritin, apiosyl liquiritin / isoliquiritin, isoliquiritin, ononin are the exclusive components of Glycyrrhiza uralensis; 3 organic acid compounds were identified, vanillic acid is the exclusive component of Angelica sinensis, glycyrrhizic acid is the exclusive component of Glycyrrhiza uralensis, ferulic acid is a common component of Angelica sinensis, Angelica pubescens and Ligusticum chuanxiong; 1 amino acid compound was identified, tryptophan is the exclusive component of Angelica sinensis; 1 saponin compound was identified, adenosine is the exclusive component of Angelica sinensis;

[0044] Using the quality detection method of the present invention, the presence and characteristics of the common characteristic peaks in the chromatogram of Juanbi Decoction can comprehensively monitor the quality of the raw materials, semi-finished products and finished products of Juanbi Decoction. By comparing the similarity degree of chromatographic characteristics, the quality of Juanbi Decoction can be evaluated, its stability and consistency can be investigated, the deficiencies of the current quality control method can be made up, and at the same time, the stability of the production process of Juanbi Decoction can be monitored to ensure the stability, uniformity and controllability of its quality;

[0045] Using the quality inspection method of the present invention, the obtained chromatogram of Juanbi Decoction improves the quality monitoring standards for the finished and semi-finished products of Juanbi Decoction, effectively preventing the occurrence of product forgery incidents, and ensuring the normal production and circulation order of Juanbi Decoction; on the basis of the present invention, the correlation research between the chromatogram information detected by the liquid chromatography-mass spectrometry method and the pharmacodynamic activity information can also be carried out, so as to deeply clarify the correlation between the internal chemical components of Juanbi Decoction and the efficacy of this preparation.

[0046] The quality inspection method described in the present invention can be used for the quality evaluation or control of the whole process of research / development / production / clinical application of Juanbi Decoction. Description of the Drawings

[0047] Figure 1 It is the UPLC-UV chromatogram and UPLC-MS total ion current chromatogram of Juanbi Decoction in Example 1 of the present invention, where from top to bottom are the UPLC-MS total ion current chromatogram (positive ion mode TIC chromatogram), the UPLC-MS total ion current chromatogram (negative ion mode TIC chromatogram), and the UPLC-UV chromatogram of Juanbi Decoction (chromatogram under the condition of ultraviolet 310 nm);

[0048] Figure 2 It is the TOF-MS first-level mass spectrum of the substances at peaks 1 to 41 of Juanbi Decoction in Example 1 of the present invention;

[0049] Figure 3 It is the MS / MS second-level mass spectrum of the substances at peaks 1 to 41 of Juanbi Decoction in Example 1 of the present invention. Detailed Embodiments

[0050] The technical solutions in the embodiments of the present invention will be described clearly and completely below. Many specific details are set forth in the following description in order to fully understand the present invention, but the present invention can also be implemented in other ways different from those described herein. Those skilled in the art can make similar promotions without departing from the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0051] Example 1 A quality inspection method for Juanbi Decoction

[0052] In this example, the reference substance of Juanbi Decoction is used for the quality inspection of Juanbi Decoction. The specific preparation method of the reference substance of Juanbi Decoction is as follows:

[0053] Take 3.75 g of Notopterygium incisum, 3.75 g of Angelica pubescens, 3.75 g of Gentiana macrophylla, 7.50 g of Piper kadsura, 2.63 g of Ligusticum chuanxiong, 1.88 g of Cinnamomum cassia, 3.00 g of Olibanum processed with vinegar, 3.00 g of Aucklandia lappa, 11.25 g of Angelica sinensis, 11.25 g of Ramulus mori and 1.88 g of Radix Glycyrrhizae Preparata, place them in a casserole, decoct twice with water. For the first time, add 10 times the amount of water, soak for 30 minutes, bring to a boil over high heat, and then decoct over low heat for 30 minutes, filter, and reserve the filtrate; for the second time, add 8 times the amount of water, bring to a boil over high heat, and then decoct over low heat for 20 minutes, filter, combine the filtrates, concentrate under reduced pressure to a concentrated solution with a ratio of the amount of crude drug input to the volume of the concentrated decoction approximately 0.6:1 (g:mL), and freeze-dry to obtain the dry extract, which is the material reference of Juanbi Decoction (i.e., dry extract powder).

[0054] This example is a quality inspection method for Juanbi Decoction, which specifically includes the following steps:

[0055] S1. Solution preparation

[0056] S11. Preparation of test solution

[0057] Take about 1 g of the material reference of Juanbi Decoction, accurately add 10 mL of 80% (v / v) methanol aqueous solution, ultrasonically treat (power 800 W, frequency 40 kHz) for 30 minutes, shake well, filter, and take the subsequent filtrate to obtain the test solution.

[0058] Meanwhile, during the preparation of the test solution, the test solution can also be prepared with the extract of Juanbi Decoction. The preparation method is as follows: Take 3.75 g of Notopterygium incisum, 3.75 g of Angelica pubescens, 3.75 g of Gentiana macrophylla, 7.50 g of Piper kadsura, 2.63 g of Ligusticum chuanxiong, 1.88 g of Cinnamomum cassia, 3.00 g of Olibanum processed with vinegar, 3.00 g of Aucklandia lappa, 11.25 g of Angelica sinensis, 11.25 g of Ramulus mori and 1.88 g of Radix Glycyrrhizae Preparata, place them in a casserole, decoct twice with water. For the first time, add 10 times the amount of water, soak for 30 minutes, bring to a boil over high heat, and then decoct over low heat for 30 minutes, filter, and reserve the filtrate; for the second time, add 8 times the amount of water, bring to a boil over high heat, and then decoct over low heat for 20 minutes, filter, combine the filtrates to obtain the extract. Accurately measure 5 mL of the extract, place it in a stoppered conical flask, accurately add 20 mL of methanol, ultrasonically treat (power 800 W, frequency 40 kHz) for 30 minutes, shake well, filter, and take the subsequent filtrate, which can be used as the test solution.

[0059] In addition, during the preparation of the test solution, the test solution can also be prepared using the concentrated solution of Juanbi Decoction. The preparation method is as follows: Take 3.75 g of Notopterygium incisum, 3.75 g of Angelica pubescens, 3.75 g of Gentiana macrophylla, 7.50 g of Piper kadsura, 2.63 g of Ligusticum chuanxiong, 1.88 g of Cinnamomum cassia, 3.00 g of Olibanum processed with vinegar, 3.00 g of Aucklandia lappa, 11.25 g of Angelica sinensis, 11.25 g of Ramulus mori, and 1.88 g of roasted Glycyrrhiza uralensis, place them in a casserole, decoct twice with water. For the first time, add 10 times the amount of water, soak for 30 minutes, bring to a boil over high heat, and then simmer for 30 minutes, filter, and reserve the filtrate; for the second time, add 8 times the amount of water, bring to a boil over high heat, and then simmer for 20 minutes, filter, combine the filtrates, and concentrate under reduced pressure to a concentrated solution with a ratio of the amount of crude drug input to the volume of the concentrated decoction approximately 0.6:1 (g:mL). Accurately measure 2 mL of the concentrated solution, place it in a stoppered conical flask, accurately add 8 mL of methanol, ultrasonically treat (power 800 W, frequency 40 kHz) for 30 minutes, shake well, filter, and take the subsequent filtrate to obtain the test solution.

[0060] S12. Preparation of Single Herb Control Material Solution

[0061] Take appropriate amounts of the powders of each herb (passed through No. 3 sieve) (wherein, about 1 g of Notopterygium incisum, about 1 g of Angelica pubescens, about 1 g of Gentiana macrophylla, about 3 g of Angelica sinensis, about 0.7 g of Ligusticum chuanxiong, about 2 g of Piper kadsura, about 0.8 g of Olibanum, about 0.5 g of Cinnamomum cassia, about 0.8 g of Aucklandia lappa, about 3 g of Ramulus mori, about 0.5 g of Glycyrrhiza uralensis), respectively place them in round-bottom flasks, add 50 mL of water, decoct for 30 minutes, let it cool, centrifuge (rotation speed 5000 revolutions per minute) for 5 minutes, take the supernatant, evaporate to dryness, dissolve the residue in 50 mL of 80 vol% methanol aqueous solution, and transfer it to a 100 mL stoppered conical flask, tightly stopper, ultrasonically treat (power 800 W, frequency 40 kHz) for 30 minutes, take out, let it cool, shake well, filter, and take the subsequent filtrate to obtain the single herb control material solutions.

[0062] S13. Preparation of Reference Substance Solution

[0063] Take an appropriate amount of ferulic acid reference substance, accurately weigh it, place it in a brown volumetric flask, add 75 vol% methanol aqueous solution to prepare a solution containing 75 μg of reference substance per 1 mL, and thus obtain the reference substance solution.

[0064] S2. Quality Inspection

[0065] Take the test solution for liquid chromatography - mass spectrometry detection to obtain the UPLC - UV chromatogram and UPLC - MS total ion current chromatogram (positive and negative ion mode TIC chromatograms) of Juanbi Decoction, which are the chromatograms of Juanbi Decoction. See specifically Figure 1 . Figure 1 The substances of chromatographic peaks numbered 1 - 41 marked in

[0066] Among them, the chromatographic conditions for liquid chromatography - mass spectrometry detection are as follows:

[0067] The chromatographic column is packed with octadecylsilyl silica gel (Waters Acquity BEH C18 chromatographic column, column length 100 mm, inner diameter 2.1 mm, particle size 1.7 μm;

[0068] The flow rate is 0.4 mL / min;

[0069] The column temperature is 40 °C;

[0070] The detection wavelength is 310 nm;

[0071] The injection volume is 3 μL;

[0072] Acetonitrile is used as mobile phase A, and 0.05 wt% formic acid aqueous solution is used as mobile phase B;

[0073] The elution mode is gradient elution, and the elution conditions are:

[0074] 0 - 12 min, 1% → 11% mobile phase A, 99% → 89% mobile phase B;

[0075] 12 - 14 min, 11% → 13% mobile phase A, 89% → 87% mobile phase B;

[0076] 14 - 15 min, 13% → 14% mobile phase A, 87% → 86% mobile phase B;

[0077] 15 - 18.5 min, 14% → 16% mobile phase A, 86% → 84% mobile phase B;

[0078] 18.5 - 21.5 min, 16% mobile phase A, 84% mobile phase B;

[0079] 21.5 - 25 min, 16% → 19% mobile phase A, 84% → 81% mobile phase B;

[0080] 25 - 26 min, 19% → 26% mobile phase A, 81% → 74% mobile phase B;

[0081] 26 - 32 min, 26% mobile phase A, 74% mobile phase B;

[0082] 32 - 33.5 min, 26% → 44% mobile phase A, 74% → 56% mobile phase B;

[0083] 33.5 - 45 min, 44% → 46% mobile phase A, 56% → 54% mobile phase B;

[0084] 45 - 46 min, 46% → 66% mobile phase A, 54% → 34% mobile phase B;

[0085] 46 - 55 min, 66% mobile phase A, 34% mobile phase B.

[0086] The mass spectrometry conditions for LC - MS detection are as follows:

[0087] The mass spectrometry detection is carried out in positive and negative ion modes respectively. The drying gas temperature is 350 °C, the drying gas flow rate is 10 L / min, the nebulizing gas pressure is 35 psi, the sheath gas temperature is 350 °C, the sheath gas flow rate is 12 L / min, and the capillary voltage is 3500 V (positive mode) and 3500 V (negative mode). The first - stage mass spectrometry selects the MS mode, and the mass scanning range is 100 - 1200 m / z. The second - stage mass spectrometry selects the Auto MSMS mode, and the collision voltage is adjusted appropriately according to different compounds. The obtained LC - MS data are collected by Agilent MassHunter (B.08.00) software. The data processing uses Agilent software Qualitative Navigator (B.08.00) and Qualitative Workflows (B.08.00).

[0088] At the same time, the LC - MS detections were carried out on each single - herb reference medicinal material solution and each reference substance solution, and the detection results were compared and analyzed with those of the test sample solution, clarifying the medicinal flavor source attribution of 41 peak substances and the qualitative analysis of the main chromatographic peaks. Among them, 10 medicinal flavors including Notopterygii Rhizoma, Gentianae Macrophyllae Radix, Angelicae Pubescentis Radix, Chuanxiong Rhizoma, Angelicae Sinensis Radix, Olibanum (processed with vinegar), Ramulus Mori, Glycyrrhizae Radix Preparata, Cinnamomi Cortex, Aucklandiae Radix, and Piperis Kadsurae Caulis can obtain the peak attribution in the mass spectrometry fingerprint of the substance reference solution.

[0089] Through the accurate mass number determination of high - resolution TOF - MS, the first - stage mass spectrometry analysis was carried out, and Figure 1 the compounds in peaks 1 - 41 were analyzed, and a total of 41 compounds were identified. The results are shown in Table 1 and Figure 2 as follows. It can be seen that in the positive mode, these compounds can generate adduct ions such as molecular ion peaks [M + H] + , [M + Na] + , [M + K] + ; while in the negative mode, they can generate adduct ions such as molecular ion peaks [M - H] - , [M + Cl] - , [M + HCOO] - . Based on this adduct ion information, the accurate molecular weight and molecular formula of the compounds can be accurately deduced (the mass determination error is ≤5 ppm, indicating that the matching results are correct), which helps with their subsequent structure identification. Among the first - stage mass spectrometry information of the 41 compound peaks in Figure 2 , most compounds have responses in both positive and negative ion modes, and some compounds can only be detected in one mode.

[0090] Further, the test solution was subjected to Auto MS / MS analysis (mainly in negative mode) under the same chromatographic conditions, that is, secondary mass spectrometry analysis, to confirm the structures of some compounds in Table 1. The results are shown in Table 1 and Figure 3 as follows. Through secondary mass spectrometry analysis, the primary mass spectrometry identification results in Table 1 were further confirmed to a certain extent.

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100] The test solution of the material reference of Juanbi Decoction was analyzed by UPLC-QTOF-MS respectively, and 41 components were detected in total. By comparing with the literature data, comparing the retention time with the reference substance, and analyzing the accurate molecular weight given by UPLC-QTOF-MS, 38 components were identified in total. And the source medicinal flavors of the compounds were attributed.

[0101] a. Coumarins

[0102] A total of 11 coumarin compounds were identified. Among them, the characteristic components of Notopterygium incisum are isomers of nodakenin, nodakenin, isomers of notopterol, notopterol; the characteristic components of Angelica pubescens are isonodakenin, osthol G / D / B, osthol A, osthol C / F / E / H, anomertin, cnidilin, columbianadin;

[0103] Furthermore, 4 compounds, namely notopterol, cnidilin, columbianadin, and nodakenin, were confirmed by comparison with the reference standard and their accurate chemical structures were determined.

[0104] b. Iridoid glycosides

[0105] A total of 5 iridoid glycosides were identified, namely loganic acid, picroside II, gentiopicroside, 6'-O-β-D-glucopyranosyl gentiopicroside, and picroside I, which were the main characteristic components. Among them, 2 compounds, gentiopicroside and loganic acid, were confirmed by comparison with reference standards and their accurate chemical structures were determined.

[0106] c. Phenylpropanoids

[0107] A total of 5 phenylpropanoids were identified. Chlorogenic acid was a common component in Angelica sinensis, Heracleum hemsleyanum, Notopterygium incisum, and Aucklandia lappa. Neochlorogenic acid was a common component in Heracleum hemsleyanum and Aucklandia lappa. Isochlorogenic acid A, isochlorogenic acid B, and isochlorogenic acid C were exclusive components of Aucklandia lappa.

[0108] d. Flavonoids

[0109] A total of 5 flavonoids were identified. Mulberroside A was an exclusive component of Ramulus Mori. Liquiritin, apiosylglycyrrhizin / isoapiosylglycyrrhizin, isoliquiritin, and ononin were exclusive components of Glycyrrhiza uralensis. And the accurate chemical structure of liquiritin was confirmed by comparison with the reference standard.

[0110] f. Organic acids

[0111] A total of 3 organic acids were identified. Vanillic acid was an exclusive component of Angelica sinensis. Glycyrrhizic acid was an exclusive component of Glycyrrhiza uralensis. Ferulic acid was a common component in Angelica sinensis, Heracleum hemsleyanum, and Ligusticum chuanxiong. And the accurate chemical structure of ferulic acid was confirmed by comparison with the reference standard.

[0112] g. Amino acids

[0113] A total of 1 amino acid was identified. Tryptophan was an exclusive component of Angelica sinensis.

[0114] h. Glycosides

[0115] A total of 1 saponin was identified. Adenosine was an exclusive component of Angelica sinensis.

[0116] The quality detection method of Juanbi Decoction in this example can be used for the quality evaluation or control of the whole process of research / development / production / clinical application of Juanbi Decoction.

[0117] Quality detection methods of Juanbi Decoction in Examples 2-7

[0118] Examples 2-7 are respectively quality detection methods of a kind of Juanbi Decoction. Their steps are basically the same as those in Example 1, except for the different process parameters. For details, see Table 2 and Table 3:

[0119] Table 2 List of process parameters in Examples 2-3

[0120]

[0121]

[0122] Table 3 List of Process Parameters in Examples 4 to 7

[0123]

[0124] The process steps and parameters of other parts in Examples 2 to 7 are the same as those in Example 1, and the obtained test results are also similar to those in Example 1, which will not be elaborated here.

[0125] Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

Claims

1. A quality detection method for Juanbi Decoction, characterized in that: The quality detection method comprises the following steps: S1. Prepare the test solution by taking Juanbi decoction; S2. Take the test solution and use high performance liquid chromatography tandem mass spectrometry to detect the quality of Juanbi decoction; The mobile phase A of the HPLC detection is acetonitrile, and the mobile phase B is a formic acid aqueous solution with a concentration of 0.04-0.06wt%; The elution method of the high performance liquid chromatography detection is gradient elution; The elution conditions of the gradient elution are: 0-12 min, 1%→11% mobile phase A, 99%→89% mobile phase B; 12-14 min, 11% → 13% mobile phase A, 89% → 87% mobile phase B; 14-15 min, 13% → 14% mobile phase A, 87% → 86% mobile phase B; 15-18.5 min, 14% → 16% mobile phase A, 86% → 84% mobile phase B; 18.5-21.5 min, 16% mobile phase A, 84% mobile phase B; 21.5-25 min, 16% → 19% mobile phase A, 84% → 81% mobile phase B; 25-26 min, 19% → 26% mobile phase A, 81% → 74% mobile phase B; 26-32 min, 26% mobile phase A, 74% mobile phase B; 32-33.5 min, 26% → 44% mobile phase A, 74% → 56% mobile phase B; 33.5-45 min, 44% → 46% mobile phase A, 56% → 54% mobile phase B; 45-46 min, 46% → 66% mobile phase A, 54% → 34% mobile phase B; 46-55min, 66% mobile phase A, 34% mobile phase B.

2. The quality detection method of Juebi Decoction according to claim 1, characterized in that: The detection wavelength of the high performance liquid chromatography detection is 290 to 330 nm.

3. The quality detection method of the Juebi Decoction according to claim 1 or 2, characterized in that: The flow rate of the high performance liquid chromatography detection is 0.38-0.45 mL / min and the column temperature is 38-45°C.

4. The quality detection method of the Juebi Decoction according to claim 1 or 2, characterized in that: The chromatographic column for high performance liquid chromatography detection uses octadecylsilane bonded silica gel as a filler.

5. The quality detection method of Juanbi Decoction according to claim 1 or 2, characterized in that: The mass spectrometry detection is performed in positive ion mode and negative ion mode respectively.

6. The quality detection method of Juanbi Decoction according to claim 1 or 2, characterized in that: The conditions for mass spectrometry detection are: drying gas temperature 345-355°C, drying gas flow rate 8-12 L / min, nebulizing gas pressure 34-36 psi, sheath gas temperature 345-355°C, sheath gas flow rate 11-13 L / min, capillary voltage in positive ion mode 3400-3600 V, and capillary voltage in negative ion mode 3400-3600 V.

7. The quality detection method of Juanbi Decoction according to claim 1 or 2, characterized in that: In the mass spectrometry detection process, the primary mass spectrometry adopts MS mode with a mass scanning range of 100 to 1200 m / z, and the secondary mass spectrometry adopts Auto MSMS mode.

8. The quality detection method of Juanbi Decoction according to claim 1 or 2, characterized in that: The test solution is prepared by adding solid Juanbi decoction to methanol aqueous solution, ultrasonically treating, and filtering; The concentration of methanol aqueous solution is 70-85 vol%; The solid form of Qunbi Decoction is a dry paste powder of Qunbi Decoction, and the weight-to-volume ratio of Qunbi Decoction to methanol aqueous solution is 0.9-1.1 g:10 mL.

9. The quality detection method of Juanbi Decoction according to claim 1 or 2, characterized in that: The test solution is prepared by taking the extract of Juanbi Decoction or the concentrated solution of Juanbi Decoction, adding methanol, ultrasonically treating, filtering and the like.

10. The quality detection method of Juanbi Decoction according to claim 9, characterized in that: The volume ratio of the Juanbi Decoction extract / Juanbi Decoction concentrate to methanol is 1:3.8-4.2.

Images