Method for identifying components of euonymus alatus decoction based on UHPLC-Q-TOF-MS technology

Abstract

The invention discloses a method for identifying components of a ramulus euonymi decoction based on a UHPLC-Q-TOF-MS technology, and relates to the technical field of medicines. The method comprises the following steps: S1, establishing a compound database; s2, extracting a sample; s3, extract detection; s4, data processing; according to the method, the euonymus alatus is subjected to water extraction through the steps from S1 to S4, the euonymus alatus water decoction is obtained, and experimental results show that the method for identifying the components of the euonymus alatus water decoction based on the UHPLC-Q-TOF-MS technology is a method for rapidly identifying the euonymus alatus water decoction or performing water extraction.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to a method for identifying components of a water decoction of Euphorbia miliariae based on UHPLC-Q-TOF-MS technology. Background Art

[0002] UHPLC-Q-TOF-MS (ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry) is a highly efficient and precise modern analytical technique that combines the high separation capabilities of ultra-high performance liquid chromatography with the precise detection advantages of high-resolution mass spectrometry. UHPLC utilizes fine-particle columns and ultra-high pressure systems to significantly improve separation efficiency, enabling efficient separation of complex samples in a short period of time. It is suitable for the analysis of complex matrices such as traditional Chinese medicine and metabolomics. Q-TOF-MS, consisting of a quadrupole mass filter and a time-of-flight mass analyzer, offers high resolution (typically >30,000), high sensitivity, and high mass accuracy (error <5 ppm), enabling precise determination of molecular weight, elemental composition, and fragment ion information of compounds. It supports both multi-stage mass spectrometry (MS / MS) and full-scan modes.

[0003] Its advantages include: ① high-throughput analysis, capable of detecting thousands of components simultaneously; ② high sensitivity, suitable for trace substance identification; ③ powerful structural analysis capabilities, enabling rapid identification of unknown compounds through fragment ion matching databases; ④ compatibility with positive and negative ion modes, expanding compound coverage;

[0004] This technology is widely used in the identification of chemical components of traditional Chinese medicine, metabolomics, environmental pollutant screening, and drug metabolism research. It plays a key role in the simultaneous analysis of multiple components in complex traditional Chinese medicine systems, providing key technical support for modern scientific analysis of traditional medicines.

[0005] Euonymus alatus (Thunb.) Siebold is a winged branch or wing-like appendage of the Euonymus family. It tastes bitter, pungent, and cold in nature. In the Shennong Bencao Jing, Euonymus alatus is recorded as a medium-grade herb under the name of "Euonymus". In modern times, the names of medicinal plants are mostly based on the morphology of the plant, and it is called "Euonymus alatus". Its chemical composition is complex, mainly including flavonoids, triterpenes, steroids, alkaloids, and organic acids. Euonymus alatus is often used to treat abdominal pain, amenorrhea, dysmenorrhea, metrorrhagia, postpartum abdominal pain due to stasis, lochia retention, hernia, rheumatic pain, sores, traumatic injuries, abdominal pain caused by worms, burns, and snake bites. Modern pharmacological studies have shown that it has the effects of lowering blood sugar and regulating lipids, cardiovascular protection, anti-inflammatory and antioxidant effects, and improving renal function. Analysis of the chemical composition of Euonymus alatus is of great significance for studying the efficacy mechanism of Euonymus alatus and the development of related drug products.

[0006] However, existing technologies make it difficult to quickly identify the components of the water decoction of P. argentea;

[0007] In response to the above problems, the inventors proposed a method for identifying the components of the water decoction of Psoralea corylifolia based on UHPLC-Q-TOF-MS technology to solve the above problems. Summary of the Invention

[0008] In order to solve the above problems, the purpose of the present invention is to provide a method for identifying the components of the water decoction of Euphorbia pulex based on UHPLC-Q-TOF-MS technology.

[0009] To solve the above technical problems, the present invention adopts the following technical solution: a method for identifying the components of the water decoction of Radix Glehniae based on UHPLC-Q-TOF-MS technology, comprising the following steps:

[0010] S1. Establishment of compound database

[0011] Acquire known compounds of the Chinese medicine Radix Glechomae by searching a database and establish a database, wherein the contents of the database include serial numbers, compound names, structural formulas and relative molecular masses;

[0012] S2. Sample extraction

[0013] The Chinese medicinal materials were extracted with ultrapure water according to the mass ratio. Five Chinese medicinal materials were weighed and extracted with ultrapure water. The extracted materials were cooled and the supernatant of the decoction was poured out for later use. The supernatant of the decoction was centrifuged and all samples were stored at low temperature.

[0014] S3. Extract detection

[0015] Chromatographic analysis was performed on a chromatographic system, chromatographic separation was achieved on an SB-C18 analytical column, and then elution was performed;

[0016] S4. Data Processing

[0017] Accurate analysis was performed using mass spectrometer software, and the positive and negative ion mode MS / MS analysis results were imported respectively. The previously established compound database was imported for comparison and the comparison results were exported. The results were screened and the compounds that met the criteria were selected.

[0018] Preferably, in S2, the ratio of the Chinese medicinal materials to ultrapure water is 1:8, 8.0 g of the five Chinese medicinal materials are weighed, 64 ml of ultrapure water is added, the heating extraction temperature is 94° C., and the time is set to 2 h;

[0019] The centrifugal treatment was performed at a speed of 1000 rpm, a temperature of 10° C., and a time of 15 min. All samples were stored at a temperature below 4° C.

[0020] Preferably, in S3, elution is performed under the following conditions: the mobile phase consists of (A) water containing 0.1% formic acid and (B) acetonitrile, the gradient program is: 1% B; 1-40% B; 40-95% B; after a running time of 3 min, it returns to 1% B, the injection volume is 2 μL, the flow rate is 0.3 ml / min, and mass spectra in both positive and negative modes are obtained, with an m / z range of 100 to 1700, an ESI ion source, a drying gas flow rate of 11 L / min, a temperature of 350°C, the capillary voltage, the nozzle voltage and the fragmentation voltage are set to 4500 V, 500 V and 140 V, respectively; the nebulizer pressure is 45 psi, and the column temperature is set to 30°C.

[0021] Preferably, in S4, the screening conditions are Width≥0.08, Height>10000, Area>100000, Ions>3, Diff≤±6.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] In the present invention, water extraction is performed on the Styracaena scoparia through steps S1-S4 to obtain the Styracaena scoparia water decoction of the present invention. Experimental results show that the method for identifying the components of the Styracaena scoparia water decoction based on UHPLC-Q-TOF-MS technology is a method for quickly identifying the Styracaena scoparia water decoction or water extraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0025] Figure 1 This is the positive ion flow diagram of the ghost arrow feather components of the present invention.

[0026] Figure 2 This is the negative ion flow diagram of the ghost arrow feather components of the present invention. DETAILED DESCRIPTION

[0027] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0028] Example: Figure 1-2As shown, the present invention provides a method for identifying the components of the water decoction of Radix Glehniae based on UHPLC-Q-TOF-MS technology, comprising the following steps:

[0029] S1. Establishment of compound database

[0030] By searching CNKI and PubMed databases, known compounds of the Chinese medicine Radix Glechomae were obtained and a database was established, wherein the contents of the database included serial numbers, compound names, structural formulas and relative molecular masses;

[0031] S2. Sample extraction

[0032] The Chinese medicinal materials were extracted with ultrapure water in a mass ratio of 1:8. 8.0 g of each of the five Chinese medicinal materials were accurately weighed and extracted with 64 ml of ultrapure water at 94°C for 2 h. The extracted materials were cooled and the supernatant of the decoction was poured out for later use. The supernatant of the decoction was centrifuged at 1000 rpm, 10°C for 15 min, and all samples were stored at 4°C.

[0033] S3. Extract detection

[0034] Chromatographic analysis was performed on an Agilent 6530UH LC-MS / MS system (Agilent Technologies, USA) with an SB-C18 analytical column (2.1×100 mm, 1.8 μm, SB-C18 Chromatographic separation was achieved on a RRHD (Agilent USA) and elution was performed under the following conditions: the mobile phase consisted of (A) water containing 0.1% formic acid and (B) acetonitrile with a gradient program of 1% B (0-2 min); 1-40% B (2-20 min); 40-95% B (20-26 min); with a post-run time of 3 min returning to 1% B. The injection volume was 2 μL, the flow rate was 0.3 ml / min, and mass spectra in both positive and negative modes were obtained with an m / z range of 100-1700. The ESI ion source was used, the drying gas (N2) flow rate was 11 L / min, the temperature was 350°C, the capillary voltage, nozzle voltage, and fragmentor voltage were set to 4500 V, 500 V, and 140 V, respectively. The nebulizer pressure was 45 psi, and the column temperature was set to 30°C.

[0035] S4. Data Processing

[0036] Accurate analysis was performed using Agilent Mass, and the positive and negative ion mode MS / MS analysis results were imported respectively. The previously established compound database was imported for comparison and the comparison results were exported. The results were screened with the conditions of Width ≥ 0.08, Height > 10000, Area > 100000, Ions > 3, and Diff (ppm) ≤ ± 6, and the compounds that met the conditions were selected.

[0037] The experimental results are as follows:

[0038] Total ion chromatograms of UHPLC-Q-TOF-MS of Psoralea corylifolia in positive and negative ion modes (see Figure 1 and Figure 2 ), combined with the primary and secondary mass spectrometry information, the components of the first ghost arrow feather were identified, and the results are shown in Table 3;

[0039]

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046] Table 1 Database of known compounds of the traditional Chinese medicine Radix Glechomae

[0047]

[0048]

[0049]

[0050] Table 2 Identification of 40 components of the water decoction of Psoralea corylifolia based on UHPLC-Q-TOF-MS technology

[0051]

[0052]

[0053]

[0054] Table 3 Identification of 40 components of the water decoction of Psoralea corylifolia based on UHPLC-Q-TOF-MS technology

[0055] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A method for identifying the components of a water decoction of Radix Glehniae based on UHPLC-Q-TOF-MS technology, characterized in that: The following steps are involved: S1. Establishment of compound database Acquire known compounds of the Chinese medicine Radix Glechomae by searching a database and establish a database, wherein the contents of the database include serial numbers, compound names, structural formulas and relative molecular masses; S2. Sample extraction The Chinese medicinal materials were extracted with ultrapure water according to the mass ratio. Five Chinese medicinal materials were weighed and extracted with ultrapure water. The extracted materials were cooled and the supernatant of the decoction was poured out for later use. The supernatant of the decoction was centrifuged and all samples were stored at low temperature. S3. Extract detection Chromatographic analysis was performed on a chromatographic system, chromatographic separation was achieved on an SB-C18 analytical column, and then elution was performed; S4. Data Processing Accurate analysis was performed using mass spectrometer software, and the positive and negative ion mode MS / MS analysis results were imported respectively. The previously established compound database was imported for comparison and the comparison results were exported. The results were screened and the compounds that met the criteria were selected.

2. The method for identifying the components of the water decoction of Radix Glehniae based on UHPLC-Q-TOF-MS technology according to claim 1, characterized in that: In S2, the ratio of the Chinese medicinal materials to ultrapure water is 1:8, 8.0 g of the five Chinese medicinal materials are weighed, 64 ml of ultrapure water is added, the heating extraction temperature is 94° C., and the time is set to 2 h; The centrifugal treatment was performed at a speed of 1000 rpm, a temperature of 10° C., and a time of 15 min. All samples were stored at a temperature below 4° C.

3. The method for identifying the components of the water decoction of Radix Glehniae based on UHPLC-Q-TOF-MS technology according to claim 1, characterized in that: In S3, elution was performed under the following conditions: the mobile phase consisted of (A) water containing 0.1% formic acid and (B) acetonitrile with a gradient program of 1% B; 1-40% B; 40-95% B; and a return to 1% B with a post-run time of 3 min. The injection volume was 2 μL, the flow rate was 0.3 ml / min, and mass spectra in both positive and negative modes were obtained with an m / z range of 100 to 1700. The ESI ion source had a drying gas flow rate of 11 L / min, the temperature was 350°C, the capillary voltage, nozzle voltage, and fragmentation voltage were set to 4500 V, 500 V, and 140 V, respectively. The nebulizer pressure was 45 psi, and the column temperature was set to 30°C.

4. The method for identifying the components of the water decoction of Radix Glehniae based on UHPLC-Q-TOF-MS technology according to claim 1, characterized in that: In S4, the screening conditions are Width≥0.08, Height>10000, Area>100000, Ions>3, Diff≤±6.

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