A method for detecting multiple components in a traditional Chinese medicine composition

Abstract

The application relates to the technical field of analysis, and particularly discloses a detection method for multiple components in a traditional Chinese medicine composition, wherein the traditional Chinese medicine composition preparation is prepared from raw medicinal materials such as Ephedra, gypsum, forsythia, radix scrophulariae, white mulberry bark, bitter apricot kernel, dried tangerine or orange peel, honeysuckle and the like; the detection method adopts ultra-high liquid chromatography tandem mass spectrometry, realizes efficient, accurate and sensitive simultaneous detection of multiple components, provides strong evidence for the research on the pharmacodynamic material basis of the traditional Chinese medicine composition, and has a wide application prospect and important practical value.

Description

Technical Field

[0001] This invention relates to the field of analytical technology, and in particular to a method for detecting multiple components in a traditional Chinese medicine composition. Background Technology

[0002] Traditional Chinese medicine (TCM), a treasure of the Chinese nation, boasts a history spanning thousands of years and plays a vital role in the prevention and treatment of diseases. TCM compositions, also known as compound TCM formulas, typically consist of multiple medicinal herbs and are characterized by complex pharmacology, diverse active ingredients, and significant therapeutic effects. However, it is precisely this complexity and diversity that makes the quality control and component detection of TCM compositions a major challenge.

[0003] In the field of traditional Chinese medicine (TCM) composition detection, traditional methods such as chemical quantification, ultraviolet-visible spectrophotometry, thin-layer chromatography, and gas chromatography, while meeting the detection needs of certain TCM components to some extent, often suffer from drawbacks such as low sensitivity, cumbersome operation, and limited applicability. Especially for TCM compositions containing multiple medicinal flavors and complex components, these traditional methods often struggle to achieve comprehensive detection and quantitative analysis of all active ingredients.

[0004] The traditional Chinese medicine composition involved in this invention consists of more than ten herbs, including ephedra, gypsum, forsythia, scutellaria, mulberry bark, bitter almond, tangerine peel, and honeysuckle. This compound has important pharmacological effects and clinical value in traditional Chinese medicine theory. However, due to the large number of chemical components in this compound and the complex interactions between them, its quality control and component detection are particularly difficult. Traditional detection methods often cannot achieve comprehensive detection and analysis of multiple chemical components, thus affecting the quality control and clinical application effects of this compound.

[0005] To address the aforementioned problems, this invention proposes a method for detecting multiple components in traditional Chinese medicine compositions based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS). This method enables the precise separation and detection of multiple chemical components in traditional Chinese medicine compositions, providing strong technical support for the quality control and clinical application of these compositions. Summary of the Invention

[0006] The purpose of this invention is to provide a method for detecting multiple components in a traditional Chinese medicine composition. This method uses ultra-high performance liquid chromatography-tandem mass spectrometry, which can achieve accurate separation and determination of 41 chemical components.

[0007] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0008] A method for detecting multiple components in a traditional Chinese medicine composition, wherein the traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ephedra 52-86 parts; Gypsum 194-324 parts; Forsythia 194-324 parts; Scutellaria baicalensis 78-130 parts; Morus alba root bark 194-324 parts; Bitter almond 78-130 parts; Peucedanum praeruptorum 78-130 parts; Pinellia ternata 78-130 parts; Tangerine peel 78-130 parts; Fritillaria cirrhosa 78-130 parts; Arctium lappa 78-130 parts; Honeysuckle 78-130 parts; Rheum palmatum 39-65 parts; Platycodon grandiflorus 46-76 parts; and Glycyrrhiza uralensis 39-65 parts.

[0009] The detection method employs ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) under the following conditions:

[0010] Liquid chromatography conditions: Column type C 18 Chromatographic column; column temperature 32-38℃; injection volume 8-12μL; flow rate 0.25-0.35mL / min; mobile phase A is an aqueous solution containing 0.1% formic acid; mobile phase B is acetonitrile containing 0.1% formic acid, gradient elution, specific gradient elution conditions are as follows:

[0011]

[0012] Mass spectrometry conditions: The detection mode was full mass spectrometry-secondary mass spectrometry mode with simultaneous scanning of positive and negative ions, and the scanning range for positive and negative ions was 100-1200 m / z; the scanning method was multiple reaction monitoring; and the data analysis was performed using Compound Discover 3.0 software.

[0013] Simultaneous scanning of positive and negative ions helps to obtain more comprehensive ion information, enabling the detection of target compounds that may exist in both positive and negative ion modes, and avoiding the omission of some important detection results due to single-mode scanning.

[0014] The full mass spectrometry-secondary mass spectrometry mode provides complete information on all ions in a sample, which helps in the initial screening and identification of complex samples. Secondary mass spectrometry obtains characteristic daughter ion information by fragmenting selected parent ions, further confirming the structure and properties of compounds and enhancing the accuracy of qualitative and quantitative analysis.

[0015] Multiple reaction monitoring (MRM) can greatly improve the sensitivity and selectivity of detection by selecting specific parent ion-daughter ion pairs for monitoring. It is especially suitable for the quantitative analysis of trace target compounds in complex samples, and can effectively reduce matrix interference and improve the reliability of analytical results.

[0016] Furthermore, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ephedra 52 parts; Gypsum 324 parts; Forsythia 194 parts; Scutellaria baicalensis 78 parts; Morus alba root bark 194 parts; Bitter almond 130 parts; Peucedanum praeruptorum 78 parts; Pinellia ternata 130 parts; Tangerine peel 78 parts; Fritillaria cirrhosa 78 parts; Arctium lappa 130 parts; Lonicera japonica 130 parts; Rheum palmatum 39 parts; Platycodon grandiflorus 76 parts; and Glycyrrhiza uralensis 65 parts.

[0017] Furthermore, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ephedra 86 parts; Gypsum 194 parts; Forsythia 324 parts; Scutellaria baicalensis 130 parts; Morus alba root bark 324 parts; Bitter almond 78 parts; Peucedanum praeruptorum 130 parts; Pinellia ternata 78 parts; Tangerine peel 130 parts; Fritillaria cirrhosa 130 parts; Arctium lappa 78 parts; Lonicera japonica 78 parts; Rheum palmatum 65 parts; Platycodon grandiflorus 46 parts; and Glycyrrhiza uralensis 39 parts.

[0018] Furthermore, the bitter almond is stir-fried bitter almond, the fritillaria is Zhejiang fritillaria, the honeysuckle is mountain honeysuckle, and the pinellia is processed pinellia.

[0019] Furthermore, the chromatographic column has dimensions of 2.1 mm × 100 mm and a packing particle size of 1.8 μm.

[0020] Furthermore, the C 18 The chromatographic column was a Waters ACQUITY UPLC HSS T3 column.

[0021] Furthermore, the method for detecting multiple components in the above-mentioned traditional Chinese medicine composition includes the following steps:

[0022] Preparation of test solution: Place the dry powder of the traditional Chinese medicine composition in a centrifuge tube, add 50% methanol aqueous solution, sonicate, take the supernatant, blow dry with nitrogen, then redissolve with 50% methanol aqueous solution, place in a centrifuge tube, centrifuge, take the supernatant, and filter with a 0.22μm microporous membrane.

[0023] Furthermore, the ultrasonic time is 25-35 minutes; the centrifugation speed is 12,000-16,000 rpm, and the time is 4-6 minutes.

[0024] Furthermore, the method for preparing the dry extract powder of the traditional Chinese medicine composition includes the following steps:

[0025] A. Weigh out the following ingredients according to the specified proportions: fritillaria, ephedra; gypsum; forsythia; scutellaria; mulberry bark; bitter almond; angelica; pinellia; tangerine peel; fritillaria; burdock fruit; honeysuckle; rhubarb; platycodon; and licorice. Set aside.

[0026] B. Take fritillaria, ephedra, forsythia, bitter almond, pinellia, burdock seed, and rhubarb, add 40-70% ethanol and reflux twice, each time for 1-4 hours. Add 8-10 times the amount for the first time and 6-9 times the amount for the second time. Combine the extracts, filter, recover the ethanol from the filtrate under reduced pressure, and concentrate to a clear extract with a relative density of 1.14-1.16 measured at 60℃. Set aside for later use.

[0027] C. Take gypsum, mulberry bark, angelica root, dried tangerine peel, honeysuckle, platycodon root, and licorice root, add water and decoct twice, each time for 1-4 hours. Add 9-11 times the amount for the first decoction and 7-9 times the amount for the second decoction. Combine the decoctions, filter, and concentrate to a clear extract with a relative density of 1.14-1.16 measured at 60℃. Combine this extract with the clear extract obtained in step B and set aside.

[0028] D. The combined extract obtained in step C is spray-dried to obtain the dry extract powder of the traditional Chinese medicine composition.

[0029] Compared with existing technologies, the detection method provided by this invention achieves efficient, accurate, and sensitive simultaneous detection of multiple components in traditional Chinese medicine compositions containing more than 10 complex medicinal flavors. It has many significant advantages and provides strong evidence for the study of the pharmacodynamic material basis of traditional Chinese medicine compositions.

[0030] This invention utilizes a Waters ACQUITY UPLC HSS T3 column, combined with an optimized gradient elution program, to achieve rapid and efficient separation of multiple components in traditional Chinese medicine compositions, significantly shortening analysis time. The simultaneous positive / negative ion scanning full-scale mass spectrometry-secondary mass spectrometry mode broadens the detection range, covering a wider range of compound types and improving the comprehensiveness of detection. Ultra-high performance liquid chromatography (UHPLC) combined with mass spectrometry detection significantly improves analytical sensitivity, enabling accurate detection even for active ingredients at extremely low concentrations. The application of multiple reaction monitoring (MRM) mode effectively reduces background noise interference, improving the accuracy of target compound detection. For complex traditional Chinese medicine compositions consisting of more than 10 medicinal ingredients, this method demonstrates powerful sample processing capabilities, enabling the simultaneous detection and identification of multiple active ingredients. Data analysis using CompoundDiscover 3.0 software achieves automated and intelligent mass spectrometry data interpretation, improving identification efficiency and accuracy. Comparison with public databases and MS fragmentation verification ensure the reliability of the identification results, providing a scientific basis for the study of traditional Chinese medicine components. Attached Figure Description

[0031] Figure 1 This is a mass spectrometry chromatogram of the traditional Chinese medicine composition of the present invention. Detailed Implementation

[0032] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0033] Example 1

[0034] An embodiment of the method for detecting multiple components in a traditional Chinese medicine composition of the present invention provides a traditional Chinese medicine composition made from the following raw materials in the following proportions: Ephedra 52g; Gypsum 324g; Forsythia 194g; Scutellaria baicalensis 78g; Morus alba root bark 194g; Bitter almond 130g; Peucedanum praeruptorum 78g; Pinellia ternata 130g; Tangerine peel 78g; Fritillaria cirrhosa 78g; Arctium lappa 130g; Lonicera japonica 130g; Rheum palmatum 39g; Platycodon grandiflorus 76g; Licorice 65g.

[0035] The preparation method of the extract of the above-mentioned traditional Chinese medicine composition is as follows:

[0036] A. Weigh out the following ingredients according to the specified proportions: fritillaria, ephedra; gypsum; forsythia; scutellaria; mulberry bark; bitter almond; angelica; pinellia; tangerine peel; fritillaria; burdock fruit; honeysuckle; rhubarb; platycodon; and licorice. Set aside.

[0037] B. Take fritillaria, ephedra, forsythia, bitter almond, pinellia, burdock seed, and rhubarb, add 50% ethanol and reflux twice, each time for 3 hours. Add 10 times the amount for the first time and 6 times the amount for the second time. Combine the extracts, filter, recover the ethanol from the filtrate under reduced pressure, and concentrate to a clear extract with a relative density of 1.15 measured at 60℃. Set aside for later use.

[0038] C. Take gypsum, mulberry bark, angelica root, dried tangerine peel, honeysuckle, platycodon root, and licorice root, add water and decoct twice, 2 hours each time. Add 10 times the amount for the first decoction and 7 times the amount for the second decoction. Combine the decoctions, filter, and concentrate to a clear extract with a relative density of 1.15 measured at 60°C. Combine this extract with the clear extract obtained in step B and set aside.

[0039] D. The combined extract obtained in step C is spray-dried to obtain a dry powder of the traditional Chinese medicine composition.

[0040] The detection method for multiple components in traditional Chinese medicine compositions, using ultra-high performance liquid chromatography-tandem mass spectrometry, is as follows:

[0041] Preparation of test solution: Take 200 mg of the dry extract powder of the traditional Chinese medicine composition, place it in a 15 mL centrifuge tube, add 10 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50), sonicate for 30 min, take the supernatant, blow dry with nitrogen, take 1 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50) to reconstitute, place in a centrifuge tube, centrifuge at 14,000 rpm for 5 min, take the supernatant and filter it through a 0.22 μm microporous membrane to obtain the test solution.

[0042] Liquid chromatography conditions: The column was a Waters ACQUITY UPLC HSS T3 column (2.1 mm × 100 mm, packing particle size 1.8 μm); column temperature was 35℃; injection volume was 10 μL; flow rate was 0.3 mL / min; mobile phase A was an aqueous solution containing 0.1% formic acid; mobile phase B was acetonitrile containing 0.1% formic acid, with gradient elution. Specific gradient elution conditions are as follows:

[0043]

[0044] Mass spectrometry conditions: Detection mode was full mass spectrometry-secondary mass spectrometry with simultaneous positive and negative ion scanning; positive and negative ion scanning range was 100-1200 m / z; scanning mode was multiple reaction monitoring; data analysis was performed using Compound Discover 3.0 software. Representative chromatograms of the traditional Chinese medicine composition are shown below. Figure 1 (Where A represents the negative ion mode and B represents the positive ion mode) As shown, 41 compounds were identified, as shown in Table 1.

[0045] Example 2

[0046] An embodiment of the method for detecting multiple components in a traditional Chinese medicine composition of the present invention provides a traditional Chinese medicine composition made from the following raw materials in the following proportions: Ephedra 86g; Gypsum 194g; Forsythia 324g; Scutellaria baicalensis 130g; Morus alba root bark 324g; Bitter almond 78g; Peucedanum praeruptorum 130g; Pinellia ternata 78g; Citrus reticulata peel 130g; Fritillaria cirrhosa 130g; Arctium lappa 78g; Lonicera japonica 78g; Rheum palmatum 65g; Platycodon grandiflorus 46g; Glycyrrhiza uralensis 39g.

[0047] The preparation method of the extract of the above-mentioned traditional Chinese medicine composition is as follows:

[0048] A. Weigh out the following ingredients according to the specified proportions: fritillaria, ephedra; gypsum; forsythia; scutellaria; mulberry bark; bitter almond; angelica; pinellia; tangerine peel; fritillaria; burdock fruit; honeysuckle; rhubarb; platycodon; and licorice. Set aside.

[0049] B. Take fritillaria, ephedra, forsythia, bitter almond, pinellia, burdock seed, and rhubarb, add 40% ethanol and reflux twice, each time for 4 hours. Add 8 times the amount for the first time and 9 times the amount for the second time. Combine the extracts, filter, recover the ethanol from the filtrate under reduced pressure, and concentrate to a clear extract with a relative density of 1.14 measured at 60℃. Set aside for later use.

[0050] C. Take gypsum, mulberry bark, angelica root, dried tangerine peel, honeysuckle, platycodon root, and licorice root, add water and decoct twice, 4 hours each time. Add 9 times the amount the first time and 7 times the amount the second time. Combine the decoctions, filter, and concentrate to a clear extract with a relative density of 1.16 measured at 60℃. Combine this extract with the clear extract obtained in step B and set aside.

[0051] D. The combined extract obtained in step C is spray-dried to obtain a dry powder of the traditional Chinese medicine composition.

[0052] The detection method for multiple components in traditional Chinese medicine compositions, using ultra-high performance liquid chromatography-tandem mass spectrometry, is as follows:

[0053] Preparation of test solution: Take 200 mg of the dry extract powder of the traditional Chinese medicine composition, place it in a 15 mL centrifuge tube, add 10 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50), sonicate for 25 min, take the supernatant, blow dry with nitrogen, take 1 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50) to reconstitute, place it in a centrifuge tube, centrifuge at 16000 rpm for 4 min, take the supernatant and filter it through a 0.22 μm microporous membrane to obtain the test solution.

[0054] Liquid chromatography conditions: The column was a Waters ACQUITY UPLC HSS T3 column (2.1 mm × 100 mm, packing particle size 1.8 μm); column temperature was 32℃; injection volume was 12 μL; flow rate was 0.35 mL / min; mobile phase A was an aqueous solution containing 0.1% formic acid; mobile phase B was acetonitrile containing 0.1% formic acid, with gradient elution. Specific gradient elution conditions are as follows:

[0055]

[0056] Mass spectrometry conditions: Detection mode was full-range mass spectrometry-secondary mass spectrometry with simultaneous positive and negative ion scanning; positive and negative ion scanning range was 100-1200 m / z; scanning mode was multiple reaction monitoring; data analysis was performed using Compound Discover 3.0 software. Forty-one compounds were identified, as shown in Table 1.

[0057] Example 3

[0058] An embodiment of the method for detecting multiple components in a traditional Chinese medicine composition of the present invention provides a traditional Chinese medicine composition made from the following raw materials in the following proportions: Ephedra 69g; Gypsum 259g; Forsythia 259g; Scutellaria baicalensis 104g; Morus alba root bark 259g; Bitter almond 104g; Peucedanum praeruptorum 104g; Pinellia ternata 104g; Tangerine peel 104g; Fritillaria cirrhosa 104g; Arctium lappa 104g; Lonicera japonica 104g; Rheum palmatum 52g; Platycodon grandiflorus 61g; Glycyrrhiza uralensis 52g.

[0059] The preparation method of the extract of the above-mentioned traditional Chinese medicine composition is as follows:

[0060] A. Weigh out the following ingredients according to the specified proportions: fritillaria, ephedra; gypsum; forsythia; scutellaria; mulberry bark; bitter almond; angelica; pinellia; tangerine peel; fritillaria; burdock fruit; honeysuckle; rhubarb; platycodon; and licorice. Set aside.

[0061] B. Take fritillaria, ephedra, forsythia, bitter almond, pinellia, burdock seed, and rhubarb, add 70% ethanol and reflux twice, one hour each time. Add 10 times the amount for the first time and 6 times the amount for the second time. Combine the extracts, filter, recover the ethanol from the filtrate under reduced pressure, and concentrate to a clear extract with a relative density of 1.16 measured at 60℃. Set aside for later use.

[0062] C. Take gypsum, mulberry bark, angelica root, dried tangerine peel, honeysuckle, platycodon root, and licorice root, add water and decoct twice, one hour each time. Add 11 times the amount the first time and 7 times the amount the second time. Combine the decoctions, filter, and concentrate to a clear extract with a relative density of 1.14 measured at 60°C. Combine this extract with the clear extract obtained in step B and set aside.

[0063] D. The combined extract obtained in step C is spray-dried to obtain a dry powder of the traditional Chinese medicine composition.

[0064] The detection method for multiple components in traditional Chinese medicine compositions, using ultra-high performance liquid chromatography-tandem mass spectrometry, is as follows:

[0065] Preparation of test solution: Take 200 mg of the dry extract powder of the traditional Chinese medicine composition, place it in a 15 mL centrifuge tube, add 10 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50), sonicate for 30 min, take the supernatant, blow dry with nitrogen, take 1 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50) to reconstitute, place in a centrifuge tube, centrifuge at 12,000 rpm for 6 min, take the supernatant and filter it through a 0.22 μm microporous membrane to obtain the test solution.

[0066] Liquid chromatography conditions: The column was a Waters ACQUITY UPLC HSS T3 column (2.1 mm × 100 mm, packing particle size 1.8 μm); column temperature was 38℃; injection volume was 8 μL; flow rate was 0.25 mL / min; mobile phase A was an aqueous solution containing 0.1% formic acid; mobile phase B was acetonitrile containing 0.1% formic acid, with gradient elution. Specific gradient elution conditions are as follows:

[0067]

[0068] Mass spectrometry conditions: Detection mode was full-range mass spectrometry-secondary mass spectrometry with simultaneous positive and negative ion scanning; positive and negative ion scanning range was 100-1200 m / z; scanning mode was multiple reaction monitoring; data analysis was performed using Compound Discover 3.0 software. Forty-one compounds were identified, as shown in Table 1.

[0069] Example 4

[0070] An embodiment of the method for detecting multiple components in a traditional Chinese medicine composition of the present invention provides a traditional Chinese medicine composition made from the following raw materials in the following proportions: Ephedra 55g; Gypsum 254g; Forsythia 318g; Scutellaria baicalensis 107g; Morus alba root bark 203g; Bitter almond 107g; Peucedanum praeruptorum 82g; Pinellia ternata 105g; Citrus reticulata peel 84g; Fritillaria cirrhosa 125g; Arctium lappa 122g; Lonicera japonica 113g; Rheum palmatum 42g; Platycodon grandiflorus 60g; Glycyrrhiza uralensis 50g.

[0071] The preparation method of the extract of the above-mentioned traditional Chinese medicine composition is as follows:

[0072] A. Weigh out the following ingredients according to the specified proportions: fritillaria, ephedra; gypsum; forsythia; scutellaria; mulberry bark; bitter almond; angelica; pinellia; tangerine peel; fritillaria; burdock fruit; honeysuckle; rhubarb; platycodon; and licorice. Set aside.

[0073] B. Take fritillaria, ephedra, forsythia, bitter almond, pinellia, burdock seed, and rhubarb, add 60% ethanol and reflux twice, each time for 2 hours. Add 9 times the amount for the first time and 7 times the amount for the second time. Combine the extracts, filter, recover the ethanol from the filtrate under reduced pressure, and concentrate to a clear extract with a relative density of 1.15 measured at 60℃. Set aside for later use.

[0074] C. Take gypsum, mulberry bark, angelica root, dried tangerine peel, honeysuckle, platycodon root, and licorice root, add water and decoct twice, 2.5 hours each time. Add 10 times the amount for the first decoction and 7 times the amount for the second decoction. Combine the decoctions, filter, and concentrate to a clear extract with a relative density of 1.14 measured at 60°C. Combine this extract with the clear extract obtained in step B and set aside.

[0075] D. The combined extract obtained in step C is spray-dried to obtain a dry powder of the traditional Chinese medicine composition.

[0076] The detection method for multiple components in traditional Chinese medicine compositions, using ultra-high performance liquid chromatography-tandem mass spectrometry, is as follows:

[0077] Preparation of test solution: Take 200 mg of the dry extract powder of the traditional Chinese medicine composition, place it in a 15 mL centrifuge tube, add 10 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50), sonicate for 35 min, take the supernatant, blow dry with nitrogen, take 1 mL of 50% methanol aqueous solution (v:v, methanol:water = 50:50) to reconstitute, place in a centrifuge tube, centrifuge at 14,000 rpm for 5 min, take the supernatant and filter it through a 0.22 μm microporous membrane to obtain the test solution.

[0078] Liquid chromatography conditions: The column was a Waters ACQUITY UPLC HSS T3 column (2.1 mm × 100 mm, packing particle size 1.8 μm); column temperature was 35℃; injection volume was 10 μL; flow rate was 0.3 mL / min; mobile phase A was an aqueous solution containing 0.1% formic acid; mobile phase B was acetonitrile containing 0.1% formic acid, with gradient elution. Specific gradient elution conditions are as follows:

[0079]

[0080] Mass spectrometry conditions: Detection mode was full-range mass spectrometry-secondary mass spectrometry with simultaneous positive and negative ion scanning; positive and negative ion scanning range was 100-1200 m / z; scanning mode was multiple reaction monitoring; data analysis was performed using Compound Discover 3.0 software. Forty-one compounds were identified, as shown in Table 1.

[0081] Table 1

[0082]

[0083]

[0084]

[0085] The UHPLC-MS / MS detection method provided by this invention has shown significant beneficial effects in detecting traditional Chinese medicine compositions composed of more than 10 kinds of medicinal ingredients. It has detected and identified 41 compounds in the traditional Chinese medicine compositions, which not only improves the efficiency and accuracy of analysis, but also enhances the scientific nature of traditional Chinese medicine quality control. It provides strong evidence for the study of the pharmacodynamic material basis of traditional Chinese medicine compositions and has broad application prospects and important practical value.

[0086] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for detecting multiple components in a traditional Chinese medicine composition, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ephedra 52-86 parts; Gypsum 194-324 parts; Forsythia 194-324 parts; Scutellaria baicalensis 78-130 parts; Morus alba root bark 194-324 parts; Bitter almond 78-130 parts; Peucedanum praeruptorum 78-130 parts; Pinellia ternata 78-130 parts; Citrus reticulata peel 78-130 parts; Fritillaria cirrhosa 78-130 parts; Arctium lappa fruit 78-130 parts; Lonicera japonica 78-130 parts; Rheum palmatum 39-65 parts; Platycodon grandiflorus 46-76 parts; and Glycyrrhiza uralensis 39-65 parts. The detection method employs ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) under the following conditions: Liquid chromatography conditions: Column type C 18 Chromatographic column; column temperature 32-38℃; injection volume 8-12μL; flow rate 0.25-0.35mL / min; mobile phase A is an aqueous solution containing 0.1% formic acid; mobile phase B is acetonitrile containing 0.1% formic acid, gradient elution, specific gradient elution conditions are as follows: Mass spectrometry conditions: The detection mode was full mass spectrometry-secondary mass spectrometry mode with simultaneous scanning of positive and negative ions, and the scanning range for positive and negative ions was 100-1200 m / z; the scanning method was multiple reaction monitoring; and the data analysis was performed using Compound Discover 3.0 software.

2. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ephedra 52 parts; Gypsum 324 parts; Forsythia 194 parts; Scutellaria baicalensis 78 parts; Mulberry bark 194 parts; Bitter almond 130 parts; Peucedanum praeruptorum 78 parts; Pinellia ternata 130 parts; Tangerine peel 78 parts; 78 portions of fritillaria; Burdock seed 130 parts; honeysuckle 130 parts; rhubarb 39 parts; platycodon 76 parts and licorice 65 parts.

3. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ephedra 86 parts; Gypsum 194 parts; Forsythia 324 parts; Scutellaria baicalensis 130 parts; Morus alba root bark 324 parts; Bitter almond 78 parts; Peucedanum praeruptorum 130 parts; Pinellia ternata 78 parts; Citrus reticulata peel 130 parts; Fritillaria cirrhosa 130 parts; Arctium lappa 78 parts; Lonicera japonica 78 parts; Rheum palmatum 65 parts; Platycodon grandiflorus 46 parts; and Glycyrrhiza uralensis 39 parts.

4. The method for detecting multiple components in a traditional Chinese medicine composition as described in any one of claims 1 to 3, characterized in that, The bitter almonds mentioned are stir-fried bitter almonds, the fritillaria is Zhejiang fritillaria, the honeysuckle is mountain honeysuckle, and the pinellia is processed pinellia.

5. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 4, characterized in that, The chromatographic column has dimensions of 2.1 mm × 100 mm and a packing particle size of 1.8 μm.

6. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 5, characterized in that, The C 18 The chromatographic column was a Waters ACQUITY UPLC HSS T3 column.

7. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 4, characterized in that, Includes the following steps: Preparation of test solution: Place the dry powder of the traditional Chinese medicine composition in a centrifuge tube, add 50% methanol aqueous solution, sonicate, take the supernatant, blow dry with nitrogen, then redissolve with 50% methanol aqueous solution, place in a centrifuge tube, centrifuge, take the supernatant, and filter with a 0.22μm microporous membrane.

8. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 7, characterized in that, The method for preparing the dry extract powder of the traditional Chinese medicine composition includes the following steps: A. Weigh out the following ingredients according to the specified proportions: fritillaria, ephedra; gypsum; forsythia; scutellaria; mulberry bark; bitter almond; angelica; pinellia; tangerine peel; fritillaria; burdock fruit; honeysuckle; rhubarb; platycodon; and licorice. Set aside. B. Take fritillaria, ephedra, forsythia, bitter almond, pinellia, burdock seed, and rhubarb, add 40-70% ethanol and reflux twice, each time for 1-4 hours. Add 8-10 times the amount for the first time and 6-9 times the amount for the second time. Combine the extracts, filter, recover the ethanol from the filtrate under reduced pressure, and concentrate to a clear extract with a relative density of 1.14-1.16 measured at 60℃. Set aside for later use. C. Take gypsum, mulberry bark, angelica root, dried tangerine peel, honeysuckle, platycodon root, and licorice root, add water and decoct twice, each time for 1-4 hours. Add 9-11 times the amount for the first decoction and 7-9 times the amount for the second decoction. Combine the decoctions, filter, and concentrate to a clear extract with a relative density of 1.14-1.16 measured at 60℃. Combine this extract with the clear extract obtained in step B and set aside. D. The combined extract obtained in step C is spray-dried to obtain the dry extract powder of the traditional Chinese medicine composition.

9. The method for detecting multiple components in a traditional Chinese medicine composition as described in claim 7, characterized in that, The ultrasonic treatment time is 25-35 minutes; the centrifugation speed is 12,000-16,000 rpm, and the time is 4-6 minutes.

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