Quality detection method and application of cortex magnoliae officinalis seven-material decoction

Abstract

The invention provides a quality detection method and application of a cortex magnoliae officinalis seven-substance decoction, and belongs to the technical field of medicine detection.The quality detection method comprises the steps that the cortex magnoliae officinalis seven-substance decoction is used for preparing a test solution, acetonitrile (A)-0.08-0.12 wt% formic acid aqueous solution (B) serves as a mobile phase, and the quality of the cortex magnoliae officinalis seven-substance decoction is detected through ultra-high performance liquid chromatography-tandem mass spectrometry; the obtained detection result can be used for quality evaluation or control of the whole process of research / development / production / clinical application of the magnolia officinalis seven-material decoction. According to the present invention, the UPLC-Q-TOF-MS method is adopted to perform qualitative analysis on the material-based chemical components of the magnolia officinalis seven-substance decoction, such that the scientific basis is provided for the material-based material fundamental research of the magnolia officinalis seven-substance decoction, and the main chemical components are clarified.

Description

Technical Field

[0001] The present invention relates to drug detection technology, and in particular to a quality detection method and application of Houpu Qiwutang Background Art

[0002] Houpu Qiwutang was first recorded in *Synopsis of the Golden Chamber*. It is composed of Magnolia officinalis, Rheum palmatum, Aurantii Fructus Immaturus, Glycyrrhiza uralensis, Ziziphus jujuba, Cinnamomum cassia, and Zingiber officinale, and has the effects of relieving muscle spasms and cold, regulating the stomach and purging the intestine. Modernly, it is also widely used clinically to treat acute pancreatitis, intestinal obstruction, indigestion, bronchial asthma, gastric volvulus and other diseases, and is a classic traditional Chinese medicine formula with a long history. In the formula, Magnolia officinalis is used as the monarch drug to promote qi and eliminate fullness, induce qi stagnation and smooth the qi movement in the fu-organs; Zingiber officinale and Cinnamomum cassia are pungent, dispersing and warming, relieving muscle spasms and cold, regulating nutrient qi and defensive qi, and Aurantii Fructus Immaturus is used to break qi and relieve stuffiness, and they are used together as ministerial drugs, interacting with Magnolia officinalis to enhance the regulation of qi movement; Rheum palmatum is used as the assistant drug to purge heat and promote defecation, and descend turbid qi; Ziziphus jujuba and Glycyrrhiza uralensis are used as guiding drugs to tonify qi and harmonize various drugs.

[0003] Houpu Qiwutang is made from seven traditional Chinese medicines. The compound components of traditional Chinese medicine are complex and contain a variety of active substances. The comprehensive analysis of its pharmacodynamic material basis has always been a difficult point in the modernization research of traditional Chinese medicine. At present, the modern research on Houpu Qiwutang is mostly focused on pharmacological mechanisms and clinical applications. Although the characteristic chromatogram of Houpu Qiwutang has been established, the number of identified characteristic peaks is relatively small, and it is impossible to comprehensively evaluate the quality of Houpu Qiwutang.

[0004] Traditional analytical methods such as high performance liquid chromatography (HPLC) are difficult to achieve synchronous and accurate identification of multiple components due to limited separation efficiency, which restricts the research on quality control and action mechanisms. Ultra-high performance liquid chromatography tandem mass spectrometry technology has high analysis efficiency and speed, can obtain accurate molecular structure information, and can obtain the primary and multi-stage mass spectrometry information of each compound, providing a good technical means for the multi-index component analysis of complex traditional Chinese medicine systems. Therefore, it is necessary to develop a method for analyzing Houpu Qiwutang by ultra-high performance liquid chromatography tandem mass spectrometry technology, so as to more comprehensively evaluate the quality of Houpu Qiwutang. Summary of the Invention

[0005] In view of the above problems, the present invention provides a quality detection method and application of Houpu Qiwutang.

[0006] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] A quality detection method of Houpu Qiwutang, the quality detection method includes the following steps:

[0008] S1. Take Houpu Qiwutang to prepare a test solution;

[0009] S2. Take the test solution and use ultra-high performance liquid chromatography tandem mass spectrometry to detect the quality of Houpu Qiwutang;

[0010] The mobile phase A for the ultra-high performance liquid chromatography (UHPLC) detection is acetonitrile, and the mobile phase B is an aqueous formic acid solution with a concentration of 0.08 - 0.12 wt%.

[0011] The elution method for the UHPLC detection is gradient elution.

[0012] The elution conditions for the gradient elution are as follows:

[0013] From 0 to 6.5 min, 10% → 20% of mobile phase A and 90% → 80% of mobile phase B;

[0014] From 6.5 to 10.5 min, 20% → 30% of mobile phase A and 80% → 70% of mobile phase B;

[0015] From 10.5 to 19 min, 30% → 70% of mobile phase A and 70% → 30% of mobile phase B;

[0016] From 19 to 21 min, 70% of mobile phase A and 30% of mobile phase B;

[0017] From 21 to 21.5 min, 70% → 10% of mobile phase A and 30% → 90% of mobile phase B;

[0018] From 21.5 to 23.5 min, 10% of mobile phase A and 90% of mobile phase B.

[0019] Furthermore, the detection wavelength for the UHPLC detection is 230 - 270 nm.

[0020] Furthermore, the flow rate for the UHPLC detection is 0.35 - 0.45 mL / min, the column temperature is 33 - 37 °C, and the injection volume is 1 - 5 μL.

[0021] Furthermore, the chromatographic column for the UHPLC detection is packed with octadecylsilyl-bonded silica gel, specifically the Waters ACQUITY UPLC HSS T3 chromatographic column, 2.1×100 mm, 1.8 μm.

[0022] Furthermore, the mass spectrometry detection is carried out in both positive ion mode and negative ion mode.

[0023] Furthermore, the conditions for the 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 3900 - 4100 V, the capillary voltage in negative ion mode is 3400 - 3600 V, the nozzle voltage in positive ion mode is 490 - 510 V, and the nozzle voltage in negative ion mode is 990 - 1100 V.

[0024] Furthermore, during the mass spectrometry detection process, the first-stage mass spectrometry adopts the MS mode with a mass scanning range of 100 - 1500 m / z; the second-stage mass spectrometry adopts the Auto-MSMS mode, and the collision voltage is appropriately adjusted according to different compounds, specifically 10, 20, and 40 V.

[0025] Furthermore, the test solution is prepared by adding Magnolia and Seven-Ingredient Decoction to methanol for dissolution, followed by filtration.

[0026] Furthermore, the weight-to-volume ratio of Magnolia and Seven-Ingredient Decoction to the prepared test solution is 0.9 - 1.1 g:10 mL.

[0027] An application of the detection result obtained by the above quality detection method in the whole-process quality evaluation or control of the research / development / production / clinical application of Magnolia and Seven-Ingredient Decoction.

[0028] The beneficial effects of the quality detection method and application of Magnolia and Seven-Ingredient Decoction of the present invention are as follows:

[0029] The present invention uses the UPLC-Q-TOF-MS method to qualitatively analyze the chemical components of the material reference of Magnolia and Seven-Ingredient Decoction, providing a scientific basis for the research on the material basis of the material reference of Magnolia and Seven-Ingredient Decoction, and simultaneously clarifying its main chemical composition;

[0030] The present invention uses the UPLC-Q-TOF-MS method to qualitatively analyze the chemical components of the reference substance of Magnolia and Seven-Ingredient Decoction. It has high analysis efficiency and speed, can obtain accurate molecular structure information in Magnolia and Seven-Ingredient 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 Magnolia and Seven-Ingredient Decoction;

[0031] The present invention uses the UPLC-Q-TOF-MS method to qualitatively analyze the chemical components of the reference substance of Magnolia and Seven-Ingredient Decoction, providing a scientific basis for the research on the material basis of Magnolia and Seven-Ingredient Decoction, and simultaneously clarifying its main chemical composition;

[0032] The quality detection method of the present invention can jointly detect all the medicinal material components in the whole prescription of Magnolia and Seven-Ingredient Decoction;

[0033] The present invention utilizes the characteristics of different physicochemical properties of different active components in Magnolia and Seven-Ingredient Decoction to change the chromatographic conditions, thereby achieving the purpose of well separating different active components in Magnolia and Seven-Ingredient Decoction;

[0034] By selecting specific chromatographic conditions and mass spectrometry conditions, the present invention can obtain a better chromatogram of Magnolia and Seven-Ingredient Decoction;

[0035] The quality detection method of the present invention also has good feasibility, stability, and reproducibility;

[0036] The quality inspection method of the present invention can detect and clarify the origin attribution of the medicinal flavors of 90 peak substances and the qualitative analysis of the main chromatographic peaks; among them, a total of 1 alkaloid component was identified as Oblongine b of 18; a total of 63 flavonoid components were identified, namely: catechin, procyanidin B2, procyanidin B2 isomer, epicatechin, procyanidin B2 isomer, p-coumaric acid-O-glucoside, Magnoloside B, epicatechin isomers (two), lonicerin, procyanidin B-O-gallate, shafuroside, liquiritin, apiosyl liquiritin, eriocitrin, liquiritigenin, catechin gallate / epicatechin gallate, neoeriocitrin, p-coumaric acid-O-galloyl-glucoside, lonicerin, hesperetin-7-O-β-D-glucoside, nobiletin-3-O-β-D-glucoside, naringin, rhoifolin, 5-hydroxyliquiritigenin-6'-acetylglucoside, hesperidin, 6-hydroxymusizin-8-O-β-D-Glucopyranoside, neohesperidin, isoliquiritifuranoside, hesperetin-7-O-β-D-glucoside, 7-methoxyl liquiritin, aurapten, neoliquiritin, neoisoliquiritin, isoliquiritigenin, glycyrrhizin B / glycyrrhizin D2, obtusin-8-O-β-D-glucopyranoside, prunasin-7-rutinoside, naringenin, syringin, 22-β-acetyl-liquiritigenin, 22-β-acetyl-liquiritigenin isomer, fortunellin, 3-21, dihydroxy-11-O-12-oleanene-29-carboxylic acid, 3-O-glucuronic acid-(1→2)–glucuronic acid, 3-21,24-trihydroxy-11,13(18)-oleanadiene-29-carboxylic acid, 3-O-glucuronic acid-(1→2)-glucuronic acid, 3-hydroxy-3',4',5,6,7,8-hexamethoxyflavone, 3-24, dihydroxy-22-O-12-oleanene-29-carboxylic acid, 3-O-glucuronic acid-(1→2)-glucuronic acid, aurapten isomer, glycyrrhisoflavone, limonin, urolignol, nobiletin, glycyrin C, demethylnobiletin, glycyrin N, 3',4',3,5,6,7,8-heptamethoxyflavone, tangeretin, glycyllin, obacunone, glycyrin I, 5-O-demethylnobiletin, glycyrin R, glycyrin H; a total of 1 phenylpropanoid glycoside component was identified as syringin; a total of 9 saponin components were identified, namely isoglycyrrhetin-7-O-apiofuranose-7,4′-O-diglucoside, uralensis saponin C, dihydroxyglycyrrhetinic acid, glycyrrhizin A3, glycyrrhizin G2, glycyrrhizic acid, glycyrrhizic acid isomer, glycyrrhizin H2, 11-hydroxyglycyrrhizic acid;A total of 7 anthraquinone components were identified, namely emodin-8-O-β-D-glucopyranoside, Cassialoin, aloe-emodin 8-O-β-D-glucoside, chrysophanol-1-O-β-D-glucopyranoside, aloe-emodin, rhein, and emodin; a total of 5 lignan components were identified, namely Magnoloside B isomer, Magnoloside A, honokiol, Randainal, and magnolol; a total of 1 organic acid component was identified, which was cinnamic acid;

[0037] Using the quality detection method of the present invention, the presence, absence and characteristics of the common characteristic peaks in the chromatogram of Magnolia Decoction can comprehensively monitor the quality of the raw materials, semi-finished products and finished products of Magnolia Decoction. By comparing the similarity degree of chromatographic characteristics, the advantages and disadvantages of Magnolia Decoction can be evaluated, its stability and consistency can be investigated, making up for the deficiencies of the current quality control methods. At the same time, the stability of the production process of Magnolia Decoction can also be monitored to ensure the stability, uniformity and controllability of its quality;

[0038] Using the quality detection method of the present invention, the chromatogram of Magnolia Decoction improves the quality monitoring standards of the finished products and semi-finished products of Magnolia Decoction, effectively preventing the occurrence of product forgery incidents and ensuring the normal production and circulation order of Magnolia Decoction; on the basis of the present invention, the correlation research between the chromatogram information detected by the UPLC-Q-TOF-MS method and the pharmacodynamic activity information can also be carried out, so as to deeply clarify the correlation between the internal chemical components of Magnolia Decoction and the curative effect of this preparation.

[0039] The quality detection 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 Magnolia Decoction. Description of the Drawings

[0040] Figure 1 It is the UPLC-UV chromatogram and UPLC-MS total ion current chromatogram of Magnolia Decoction in Example 1 of the present invention, where from top to bottom are the UPLC-MS total ion current chromatogram (UPLC-MS-TIC diagram in positive ion mode), UPLC-MS total ion current chromatogram (UPLC-MS-TIC diagram in negative ion mode) and the UPLC-UV chromatogram of Magnolia Decoction (chromatogram under the condition of ultraviolet 250nm). Detailed Embodiments

[0041] The technical solutions in the embodiments of the present invention will be clearly and completely described 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.

[0042] Example 1 Quality Detection Method and Application of Houpu Qiwutang

[0043] In this example, the quality of Houpu Qiwutang was detected using the reference substance of Houpu Qiwutang. The specific preparation method of the reference substance of Houpu Qiwutang is as follows:

[0044] Take Magnoliae Officinalis Cortex, Glycyrrhizae Radix et Rhizoma, Rhei Radix et Rhizoma, Jujubae Fructus, Aurantii Fructus Immaturus, Cinnamomi Ramulus, and Zingiberis Rhizoma Recens, a total of seven herbs. Cut Zingiberis Rhizoma Recens into thick slices and break Jujubae Fructus. Add water and heat with an open flame. Bring to a boil over high heat and then simmer for 25 minutes. Filter, concentrate under reduced pressure, and freeze-dry the resulting concentrated solution to obtain the reference substance of Houpu Qiwutang (i.e., dry extract powder).

[0045] This example is a quality detection method and application of Houpu Qiwutang, specifically including the following steps:

[0046] S1. Solution Preparation

[0047] S11. Preparation of Test Solution

[0048] Take about 2.5 g of the reference substance of Houpu Qiwutang, weigh accurately, place it in a stoppered conical flask, accurately add 25 mL of methanol, stopper tightly, weigh, ultrasonically treat (power 500 W, frequency 40 kHz) for 30 minutes, let it cool, weigh again, make up the lost weight with methanol, shake well, filter, and take the subsequent filtrate to obtain the test solution.

[0049] S12. Preparation of Single Herb Solution (the herbs that can be attributed in the fingerprint)

[0050] Weigh 1.44 g of Magnoliae Officinalis Cortex, 0.675 g of Aurantii Fructus Immaturus, 0.54 g of Rhei Radix et Rhizoma, 0.36 g of Cinnamomi Ramulus, and 0.54 g of Glycyrrhizae Radix et Rhizoma. Add 40 mL of water to each respectively, decoct for 25 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 5 mL of methanol by ultrasonic treatment, transfer it to a 5-mL volumetric flask, make up to the mark with methanol, shake well, filter, and take the subsequent filtrate to obtain the single herb solution corresponding to each herb.

[0051] S2. Quality Detection

[0052] Take the test solution for UPLC-Q-TOF-MS detection to obtain the UPLC-UV chromatogram and UPLC-MS total ion current chromatogram (UPLC-MS-TIC chromatograms in positive and negative ion modes) of Houpu Qiwutang, which is the chromatogram of Houpu Qiwutang. See specifically Figure 1 . Figure 1 For the substances of chromatographic peaks numbered 1 - 90 marked in

[0053] At the same time, perform UPLC-Q-TOF-MS detection on each single herb solution and compare and analyze the detection results with those of the test solution.

[0054] Among them, the instrument and equipment for UPLC-Q-TOF-MS detection are as follows:

[0055] Agilent 1290 Infinity II ultra-high performance liquid chromatography (UPLC), connected to a G6530C quadrupole-time-of-flight tandem mass spectrometer (Q-TOF-MS). Equipped with an independent quaternary pump, an autosampler, a column oven, a diode array detector (DAD), and an electrospray ionization source (ESI).

[0056] The chromatographic conditions for UPLC-Q-TOF-MS detection are as follows:

[0057] Chromatographic separation is carried out using a Waters ACQUITY UPLC HSS T3 chromatographic column (2.1×100 mm, 1.8 μm), equipped with an online filter;

[0058] The column temperature is 35 °C;

[0059] The flow rate is 0.4 mL / min;

[0060] The detection wavelength is 250 nm;

[0061] The injection volume is 2 μL;

[0062] Acetonitrile is used as mobile phase A, and a 0.1 wt% formic acid aqueous solution is used as mobile phase B;

[0063] The elution method is gradient elution, and the elution conditions are as follows:

[0064] 0 - 6.5 min, 10% → 20% mobile phase A, 90% → 80% mobile phase B;

[0065] 6.5 - 10.5 min, 20% → 30% mobile phase A, 80% → 70% mobile phase B;

[0066] 10.5 - 19 min, 30% → 70% mobile phase A, 70% → 30% mobile phase B;

[0067] 19 - 21 min, 70% mobile phase A, 30% mobile phase B;

[0068] 21 - 21.5 min, 70% → 10% mobile phase A, 30% → 90% mobile phase B;

[0069] 21.5 - 23.5 min, 10% mobile phase A, 90% mobile phase B.

[0070] The mass spectrometry conditions for UPLC-Q-TOF-MS detection are as follows:

[0071] Mass spectrometry detection was performed in positive and negative ion modes, respectively, with a drying gas temperature of 350°C, a drying gas flow rate of 10 L / min, a nebulizer gas pressure of 35 psi, a sheath gas temperature of 350°C, a sheath gas flow rate of 12 L / min, a capillary voltage of 4000 V (positive ion mode) and 3500 V (negative ion mode), and a nozzle voltage of 500 V (positive ion mode) and 1000 V (negative ion mode). The primary mass spectrometry used the MS mode, with a mass scan range of 100-1500 m / z. The secondary mass spectrometry used the Auto-MSMS mode, with collision voltages of 10, 20, and 40 V. The obtained LC-MS data were collected by Agilent MassHunter (B.08.00) software. Data processing was performed using Agilent software Qualitative Navigator (B.08.00) and Qualitative Workflows (B.08.00).

[0072] Analysis of test results: The material benchmark of Houpu Qiwu Decoction is composed of seven medicinal flavors, namely, Magnolia officinalis, Licorice, Rhubarb, Jujube, Citrus aurantium, Cinnamomum cassia and ginger. By comparing and analyzing the material benchmark with the liquid mass spectrometry test results of each component single medicine, the source of the medicinal flavors of 90 peak substances and the qualitative identification of the main chromatographic peaks were clarified. The medicinal flavors that can be attributed in the liquid mass fingerprint are Magnolia officinalis, Licorice, Rhubarb, Citrus aurantium and Cinnamomum cassia. Therefore, among the seven medicinal flavors in the prescription, five medicinal flavors can obtain the peak attribution in the material benchmark liquid mass fingerprint.

[0073] The peaks 1-90 were identified by high-resolution TOF-MS accurate mass determination. The primary mass spectrometry results confirmed the primary mass spectrometry information of 90 compound peaks, most of which responded in both positive and negative ion modes, and some compounds were only detected in one mode. The results are shown in Table 1. It can be seen that these compounds can generate [M+H] in the positive mode. + 、[M+Na] + In negative mode, molecular ion peaks [MH] are generated. - and [M+Cl] - 、[M+HCOO] - Based on the information of these adduct ions, the precise molecular weight and molecular formula of the compound can be accurately inferred (the mass measurement error is ≤5ppm, indicating that the matching result is correct), which is helpful for its subsequent structural identification.

[0074] Furthermore, the test solution was subjected to Auto-MS / MS analysis (positive and negative modes) under the same chromatographic conditions. One of the appropriate ionization modes was selected to confirm the structure of peak substances 1-90, and the results are shown in Table 1. It can be seen that the secondary mass spectrometry analysis confirmed to a certain extent that

[0075] The identification results of the first-level mass spectrometry in Table 1.

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093] The test solution of the reference substance of Houpu Qiwu Decoction was analyzed by UPLC-Q-TOF-MS, and a total of 90 components were detected. By comparing with the literature data and analyzing the accurate molecular weights given by UPLC-Q-TOF-MS, 88 of them were identified. And the source medicinal flavors of the compounds were attributed.

[0094] a. Alkaloids

[0095] A total of 1 alkaloid component was identified, which is Oblongine b with peak number 18.

[0096] b. Flavonoids

[0097] A total of 63 flavonoid components were identified, which are:

[0098] Catechin with peak number 1, Procyanidin B2 with peak number 2, Procyanidin B2 isomer with peak number 3, Epicatechin with peak number 5, Procyanidin B2 isomer with peak number 7, p-Coumaric acid-O-glucoside with peak number 9, Magnoloside B with peak number 10, Epicatechin isomers with peak numbers 11 and 12, Lonicerin with peak number 14, Procyanidin B-O-gallate with peak number 15, Schaftoside with peak number 19, Liquiritin with peak number 20, Apiosyl Liquiritin with peak number 21, Eriodictyol glucoside with peak number 22, Liquiritigenin with peak number 23, Catechin gallate / Epicatechin gallate with peak number 25, Homoeriodictyol glucoside with peak number 26, p-Coumaric acid-O-galloyl-glucoside with peak number 27, Lonicerin glycoside with peak number 28, Hesperetin-7-O-β-D-glucoside with peak number 30, Nobiletin-3-O-β-D-glucoside with peak number 31, Naringin with peak number 32, Rhoifolin with peak number 33, 5-Hydroxyliquiritigenin-6'-acetylglucoside with peak number 34, Hesperidin with peak number 35, 6-hydroxymusizin-8-O-β-D-Glucopyranoside with peak number 36, Neohesperidin with peak number 37, Isoliquiritifuranoside with peak number 38, Hesperetin-7-O-β-D-glucoside with peak number 39, 7-Methoxyl Liquiritin with peak number 40, Aurapten with peak number 41, Neo-Liquiritin with peak number 42, Neo-Isoliquiritin with peak number 43, Isoliquiritigenin with peak number 44, Glycoside B / Glycoside D2 of Liquiritin with peak number 45, Cassia ketone-8-O-β-D-glucopyranoside with peak number 47, Prunasin-7-rutinoside with peak number 48, Naringenin with peak number 53, Syringin with peak number 55, 22-β-Acetyl-Liquiritigenin with peak number 56, 22-β-Acetyl-Liquiritigenin isomer with peak number 57, Fortunellin with peak number 59, 3-21, Dihydroxy-11-O-12-oleanene-29-carboxylic acid, 3-O-glucuronic acid-(1→2)–glucuronic acid with peak number 60, 3-21,24-Trihydroxy-11,13(18)-oleanadiene-29-carboxylic acid, 3-O-glucuronic acid-(1→2)-glucuronic acid with peak number 61, 3-Hydroxy-3',4',5,6,7,8-hexamethoxyflavone with peak number 62, 3-24, Dihydroxy-22-O-12-oleanene-29-carboxylic acid, 3-O-glucuronic acid-(1→2)-glucuronic acid with peak number 63, Aurapten isomer with peak number 64, Glycycoumarin with peak number 67, Limonin with peak number 68, Uralenol with peak number 69, Nobiletin with peak number 74, Glycyrrhenin C with peak number 75, Demethylnobiletin with peak number 77, Glycyrrhenin N with peak number 78, 3',4',3,5,6,7,8-Hepamethoxyflavone, nobiletin with peak number 80, liquiritigenin with peak number 81, obacunone with peak number 82, glycyrrhenin I with peak number 83, 5-O-demethylnobiletin with peak number 85, liquiritin R with peak number 89, liquiritin H with peak number 90.,

[0099] Naringin, neohesperidin, and liquiritin were identified by reference substances.

[0100] c. Phenylpropanoid glycosides

[0101] A total of 1 phenylpropanoid glycoside component was identified, which is syringin with peak number 4.

[0102] d. Saponins

[0103] A total of 9 saponin components were identified, namely:

[0104] Isoliquiritigenin-7-O-apiosyl-7,4′-O-diglucoside with peak number 8, Glycyrrhizasaponin C with peak number 51, Dihydroxyglycyrrhetinic acid with peak number 52, Glycyrrhizasaponin A3 with peak number 54, Glycyrrhizasaponin G2 with peak number 58, Glycyrrhizic acid with peak number 65, Glycyrrhizic acid isomer with peak number 71, Glycyrrhizasaponin H2 with peak number 73, 11-Hydroxyglycyrrhizic acid with peak number 76.

[0105] And glycyrrhizic acid was identified by reference substance.

[0106] e. Anthraquinones

[0107] A total of 7 anthraquinone components were identified, namely:

[0108] Emodin-8-O-β-D-glucopyranoside with peak number 24, Cassialoin with peak number 29, Aloe-emodin 8-O-β-D-glucoside with peak number 49, Chrysophanol-1-O-β-D-glucopyranoside with peak number 50, Aloe-emodin with peak number 66, Rhein with peak number 70, Emodin with peak number 84.

[0109] And rhein was identified by reference substance.

[0110] f. Lignans

[0111] A total of 5 lignan components were identified, namely:

[0112] Magnoloside B isomer with peak number 16, Magnoloside A with peak number 17, Honokiol with peak number 86, Randainal with peak number 87, Magnolol with peak number 88.

[0113] And magnolol and honokiol were identified by reference substances.

[0114] g. Organic acids

[0115] A total of 1 organic acid component was identified, which is cinnamic acid with peak number 46.

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

[0117] The liquid chromatography-mass spectrometry fingerprint (including but not limited to UPLC-UV chromatogram and / or UPLC-MS total ion current chromatogram), primary analysis results (such as Table 1 in the example), and secondary mass spectrometry analysis results (such as Table 1 in the example) obtained by the quality detection method in this example can also be respectively used for the quality evaluation or control of the whole process of research / development / production / clinical application of Magnolia and Seven-Ingredient Decoction. For example, the peak number and retention time can be directly used for the quality evaluation or control of the whole process of research / development / production / clinical application of Magnolia and Seven-Ingredient Decoction.

[0118] Quality Detection Method and Application of Magnolia and Seven-Ingredient Decoction in Examples 2 - 5

[0119] Examples 2 - 5 are respectively a quality detection method and application of Magnolia and Seven-Ingredient Decoction. Their steps are basically the same as those in Example 1, with the only difference being the process parameters. For details, see Table 2:

[0120] Table 2 List of Process Parameters in Examples 2 - 5

[0121]

[0122]

[0123] The process steps and parameters of other parts in Examples 2 - 5 are the same as those in Example 1, and the obtained detection results are also similar to those in Example 1, which will not be elaborated here.

[0124] Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

Claims

1. A quality detection method for Houpu Qiwu Decoction, characterized in that: The quality detection method comprises the following steps: S1. Prepare the test solution with Houpo Qiwu decoction; S2. Take the test solution and use ultra-high performance liquid chromatography tandem mass spectrometry to detect the quality of Houpu Qiwu Decoction; The mobile phase A of the ultra-high performance liquid chromatography detection is acetonitrile, and the mobile phase B is a formic acid aqueous solution with a concentration of 0.08-0.12 wt%; The elution mode of the ultra-high performance liquid chromatography detection is gradient elution; The elution conditions of the gradient elution are: 0-6.5 min, 10%→20% mobile phase A, 90%→80% mobile phase B; 6.5-10.5 min, 20%→30% mobile phase A, 80%→70% mobile phase B; 10.5-19 min, 30%→70% mobile phase A, 70%→30% mobile phase B; 19-21 min, 70% mobile phase A, 30% mobile phase B; 21-21.5 min, 70% → 10% mobile phase A, 30% → 90% mobile phase B; 21.5~23.5min, 10% mobile phase A, 90% mobile phase B.

2. The quality detection method of Houpo Qiwu Decoction according to claim 1, characterized in that: The detection wavelength of the ultra-high performance liquid chromatography detection is 230-270 nm.

3. The quality detection method of Houpo Qiwu Decoction according to claim 1 or 2, characterized in that: The flow rate of the ultra-high performance liquid chromatography detection is 0.35-0.45 mL / min and the column temperature is 33-37°C.

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

5. The quality detection method of Houpo Qiwu 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 Houpo Qiwu 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 3900-4100 V, capillary voltage in negative ion mode 3400-3600 V, nozzle voltage in positive ion mode 490-510 V, and nozzle voltage in negative ion mode 990-1100 V.

7. The quality detection method of Houpo Qiwu 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 1500 m / z; the secondary mass spectrometry adopts Auto-MSMS mode.

8. The quality detection method of Houpo Qiwu Decoction according to claim 1 or 2, characterized in that: The test solution was prepared by dissolving Houpo Qiwu decoction in methanol and filtering it.

9. The quality detection method of Houpo Qiwu Decoction according to claim 8, characterized in that: The weight-to-volume ratio of Houpo Qiwu Decoction to the prepared test sample solution is 0.9-1.1 g:10 mL.

10. Application of the test result obtained by the quality testing method according to any one of claims 1 to 9 in the quality evaluation or control of the whole process of research / development / production / clinical application of Houpo Qiwu Decoction.

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