CN109444275B - Method for rapidly screening medicinal materials of wisteria floribunda and sinomenium acutum - Google Patents
Method for rapidly screening medicinal materials of wisteria floribunda and sinomenium acutum Download PDFInfo
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Abstract
The invention discloses a UPLC-MS-MS method for rapidly screening wisteria floribunda and sinomenium cyrtonema, which comprises the following steps: pulverizing radix tetrastigme medicinal materials to be distinguished, sieving to prepare a test sample, adding 10-20 times of 50-70% ethanol, carrying out ultrasonic extraction for 20-40 minutes to obtain a test sample solution, then carrying out UPLC-MS-MS system analysis to obtain mass spectrum data corresponding to the test sample, carrying out data analysis, and carrying out chromatographic peak extraction and mass spectrum information characterization on the test sample; and judging whether the test sample contains the specific flavone characteristic compound of the trefoil of wisteria according to the chromatographic information and the mass spectrum information of the test sample. The method has the advantages of rapid screening and high accuracy, and greatly simplifies and improves the workload in the analysis process of different germplasms of radix tetrastigme (wisteria floribunda and sinomenium cyrtonema).
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine analysis and research, and particularly relates to a method for rapidly screening medicinal materials including Chinese wistaria and sinomenium cyrtonema by using a flavone as a characteristic compound and applying UPLC-MS/MS.
Background
Radix tetrastigme is a peculiar rare medicinal plant in China, is Tetrastigma hemsleyanum Diels et Gilg (Tetrastigma hemsleyanum Diels) of the genus Tetrastigma in the family Vitaceae, can be used as a medicine by all herbs, and has the best medicinal effect on underground root tubers. It is cool in nature, pungent and bitter in taste, and nontoxic, has effects of protecting liver, resisting virus, resisting inflammation, relieving pain, and resisting tumor, and also has significant curative effect on measles complicated pneumonia, infantile hyperpyrexia, tonsillitis, etc. And can be widely used in various Chinese medicinal preparations or health products, such as HUATUOFENGTONGBAO, JIEZHUKANG Capsule, and JINQI tablet.
The producing areas of radix tetrastigme are mainly distributed in Zhejiang, Jiangxi, Fujian, Hubei, Hunan, Guangdong, Sichuan and other areas. The germplasm types of radix tetrastigme are mainly distinguished from leaf size and shape, surface leather degree and leaf edge sawtooth, vine color and shape, and are generally roughly divided into a Chinese wisteria and a Chinese ivy, and the two have larger difference in form, but the medicinal material forms are difficult to distinguish. The new vine of the wisteria floribunda (also known as Zhejiang radix tetrastigme) is green, and the old vine at the lower part of the plant gradually becomes purple or brown after 2-3 years. The vine is fine, has edges and corners, has small leaves, has a calabash shape with roots, and is mainly distributed in Zhejiang, Jiangxi and other places. The surface of the caulis sinomenii radix tetrastigme (also called Guangxi radix tetrastigme) old rattan is bluish white, the vine is thicker, the leaf is larger, the root tuber is larger, and the old rattan is mainly distributed in partial areas of Hunan, Fujian, Guangxi and Yunnan of China. In contrast, the sinomenium acutum has wide distribution region, vigorous growth, high yield and large quantity. However, the radix tetrastigme of the sinomenium acutum is generally considered to be cold in root tuber and inferior to the radix tetrastigme of the sinomenium acutum in medicinal efficacy, and the radix tetrastigme produced in Zhejiang is traditionally used as a genuine medicinal material, so the price of the radix tetrastigme of the sinomenium acutum in the market is far higher than that of the radix tetrastigme of the sinomenium acutum.
The chemical components in the radix tetrastigme comprise flavone, flavonoid glycoside, starch, reducing sugar, steroid compounds, amino acid and the like, wherein the flavone and the glycoside thereof are important active substances. A great deal of radix tetrastigme pharmacological research on radix tetrastigme tuberous roots at present shows that the antitumor effective substances of the radix tetrastigme tuberous roots are flavonoid components. Due to the fact that the market demand is larger and larger, the price of medicinal materials is continuously increased, however, the wild Chinese wistaria root is in short supply, so that the wild resource is over-mined, and serious damage is caused. The artificial cultivation difficulty is high, the underground root tuber of the medicinal part of the medicinal plant grows slowly, and the requirement of commercial medicinal materials can be met in 3-5 years. And the underground root tuber yield and the effective component content of different production areas and different germplasm sources (the wisteria floribunda and the sinomenium floribunda) are obviously different. At present, the radix tetrastigme germplasm in the medicinal material market is mixed, and the price difference of medicinal materials from different germplasm sources (wisteria floribunda and sinomenium tetragonolobum) is very different. Therefore, the phenomenon that the cheap common counterfeit Chinese yam rhizome is used as the authentic Chinese wistaria leaf to sell so as to obtain the violence appears in the market, and the safety and the effectiveness of the medication are difficult to ensure.
The acceleration of the modernization pace of traditional Chinese medicine puts forward a new requirement on standardization of traditional Chinese medicine, and the standardized production is the key for the development of the traditional Chinese medicine industry and the entry of the traditional Chinese medicine industry into the international market and is also the working core of the GAP (Good agricultural Practice, the Chinese medicinal material production quality management standard) which is vigorously promoted at present. The germplasm standard and scientific identification method are the premise of quality standard control and management, so that the method for identifying the traditional Chinese medicinal materials needs to meet the requirements of rapidness, accuracy, standardization and the like. However, the existing identification method of the radix tetrastigme is mainly based on morphological and conventional chemical component analysis, even only by empirical judgment, and no research report for establishing a rapid identification technology of the wisteria floribunda and the radix tetrastigme by using a modern analysis method is seen. Has very important practical significance for standardizing the radix tetrastigme market, ensuring the clinical curative effect and protecting the benefits of consumers. Therefore, there is a need to develop a method for identifying different germplasm radix tetrastigme with high practicability, and solving the problems of mixed true and false and uneven quality in the medicinal material market.
At present, the research on chemical components in the radix tetrastigme is reported, but the germplasm identification and quality control by taking a definite component as an index are not reported. In recent years, with the development of analysis technology, Ultra Performance Liquid Chromatography (UPLC) tandem triple quadrupole mass spectrometry (TQD-MS/MS) is widely used in the analysis of traditional Chinese medicines and compound medicines due to its fast separation speed, high sensitivity and strong measurement accuracy.
At present, the identification methods for traditional Chinese medicine products mainly comprise PCR, IR, NIR and the like, and a large number of false positive and false negative problems exist in the methods, so that important substance information can be omitted when the methods are used for analysis, the screened result cannot be effectively identified, and the identification of the same traditional Chinese medicine with different germplasms is difficult. The authenticity and the producing area of the radix tetrastigme are identified by various methods, such as: the PCR-RFLP method (ZL201510016355.4) for rapidly identifying the radix tetrastigme and various pseudo-mixed products thereof, the method (ZL201310373809.4) for identifying the medicinal plant radix tetrastigme, the method (201710762908.X) for identifying the origin of the radix tetrastigme and the method (201710371389.4) for rapidly detecting the origin of the radix tetrastigme by near infrared spectroscopy, however, the method for effectively identifying and distinguishing different germplasm radix tetrastigme (wisteria zizanoides and sinomenium acutanguticum) medicinal materials is not available.
Disclosure of Invention
The invention aims to provide a rapid and high-accuracy UPLC-MS-MS method for rapidly screening medicinal materials of the wisteria floribunda and the sinomenium cyrtonema, so that the workload in the analysis process of different germplasms of the sinomenium cyrtonema and the sinomenium cyrtonema is greatly simplified and improved.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for rapidly screening medicinal materials of wisteria floribunda and sinomenium acutum comprises the following steps:
1) pulverizing radix tetrastigme medicinal materials to be distinguished, and sieving to prepare a sample to be tested;
2) taking the sample to be tested in the step 1), adding 10-20 times of 50-70% ethanol, carrying out ultrasonic extraction for 20-40 minutes to obtain a sample solution to be tested, and then carrying out UPLC-MS-MS system analysis to obtain mass spectrum data corresponding to the sample to be tested;
3) directly importing the data acquired in the step 2) into mass spectrometry software, then carrying out data analysis, and carrying out chromatographic peak extraction and mass spectrometry information characterization on a sample to be tested;
4) judging whether the sample contains kaempferol-3-O-beta-D-furanose-7-O-alpha-L-rhamnoside and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside or not according to the chromatographic information and mass spectrum information of the sample obtained in the step 3); if the composition contains kaempferol-3-O-beta-D-furanosyl-7-O-alpha-L-rhamnoside and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside, the composition is radix Hemsleyae Macrospermae; if not, the radix tetrastigme is the caulis sinomenii.
Preferably, in the step 4), whether the sample contains kaempferol-3-O-beta-D-furanose-7-O-alpha-L-rhamnoside and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside is judged according to the peak area of chromatogram, the mass number of an excimer peak corresponding to the peak retention time and the information of a secondary mass spectrum.
The kaempferol-3-O-beta-D-furanose-7-O-alpha-L-rhamnoside in the step 4): molecular weight of 726, molecular formula C32H38O19The secondary mass fragment was 647.5,527.4,486.6,431.9,392.1,289.7.
The kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside in the step 4): molecular weight is 594, molecular formula C27H30O15The secondary mass fragment was 515.5,451.7,386.5,291.0,246.5.
Preferably, in the step 1), the test sample is pulverized and sieved by a 60-mesh sieve, and is stored in a dark place at the temperature of minus 20 ℃;
preferably, in the step 2), 1.0g of the sample to be tested in the step 1) is taken, precisely weighed, placed in a conical flask with a plug, precisely added with 10ml of 50% ethanol, weighed, ultrasonically treated for 30 minutes, cooled, weighed again, complemented with 50% ethanol for weight loss, shaken up, centrifuged, precisely weighed with 5ml of supernatant, placed in a brown measuring flask with 25ml, added with 50% ethanol to the scale, shaken up, filtered, and a subsequent filtrate is taken for filtration to obtain a sample solution to be tested.
Preferably, in step 3), the mass spectrometry software is MassLynx V4.1 software.
Preferably, in step 2), the analysis conditions of the UPLC-MS system are:
chromatographic conditions are as follows: waters ACQUITY UPLC BEH C18 column, 2.1X 100mm, 1.7 μm; using 0.1% formic acid-water as mobile phase a and acetonitrile as mobile phase B, gradient elution is shown in the following table:
flow rate: 0.3 ml/min; the column temperature is 30 ℃; the detection wavelength is 254 nm;
mass spectrum conditions:
MS system ACQUITY UPLC-TQD-MS triple quadrupole mass spectrometer; the conditions of use of the tandem mass spectrometer were: ionization mode ESI (-); the detection mode is as follows: detecting multiple reactions; capillary voltage: 3.0 Kv; taper hole voltage: 38V; the source temperature is 120 ℃; the temperature of the desolvation: 350 ℃; desolventizing flow rate: 500L/hr; taper hole air flow rate: 50L/hr; collision energy: 19 ev; residence time: 0.2 s; full scanning: the scan surface has a molecular weight of 100 to 1000.
Compared with the prior art, the invention has the following beneficial effects:
(1) the method for rapidly screening the wisteria floribunda and the sinomenium tricuspidatum by using UPLC-MS-MS for the first time has the advantages of rapidness, accuracy, stability and the like; therefore, the workload of different germplasms (including the wisteria floribunda and the sinomenium floribundum) of the tetrastigma hemsleyanum is greatly simplified and improved, and meanwhile, the rapid classification and identification of complex components are facilitated through a diagnostic fragment filtering strategy and the auxiliary analysis of other fragments and neutral lost molecules. The technical problem of the method for identifying and distinguishing the trefoil of the wisteria floribunda and the trefoil of the sinomenium lyratum is effectively solved by establishing a fragment diagnosis filtering strategy and a neutral loss filtering strategy and quickly screening the trefoil of the wisteria floribunda and the sinomenium cyrtonema.
(2) The preparation method adopts the test sample of the medicinal materials of the wisteria florbunda and the sinomenium cyrtophyllum as the test sample, and researches the optimization investigation of the extraction process to obtain the preparation process of the test sample of the wisteria florda and the sinomenium cyrtophyllum; the screening method of the wisteria floribunda and sinomenium acutum with different germplasms established by the invention can be popularized and applied to the raw material selection and preparation processes of decoction pieces of wisteria floribunda and sinomenium acutum and related preparations thereof.
Drawings
FIG. 1 is a UPLC comparison chart of a test sample of wisteria floribunda and a test sample of sinomenium floribundum;
FIG. 2 is a TIC comparison graph of a test sample of radix tetrastigme of wisteria floribunda and a test sample of radix tetrastigme of sinomenium tetrandra;
FIG. 3 is a structural diagram of main flavone components in the wisteria florbunda;
FIG. 4 is a structural diagram of the main flavone components in the tetrastigma hemsleyanum of Sinomenium acutum;
FIG. 5 is the PCA diagram of the principal component analysis of the Hemsleya amabilis Hemsleya and Hemsleya amabilis Hemsleya test sample.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
A method for rapidly screening medicinal materials of wisteria floribunda and sinomenium acutum comprises the following steps:
1) pulverizing radix tetrastigme medicinal materials to be distinguished, and sieving to prepare a sample to be tested;
2) taking the sample to be tested in the step 1), adding 10-20 times of 50-70% ethanol, carrying out ultrasonic extraction for 20-40 minutes to obtain a sample solution to be tested, and then carrying out UPLC-MS-MS system analysis to obtain mass spectrum data corresponding to the sample to be tested;
3) directly importing the data acquired in the step 2) into mass spectrometry software, then carrying out data analysis, and carrying out chromatographic peak extraction and mass spectrometry information characterization on a sample to be tested;
4) judging whether the sample contains kaempferol-3-O-beta-D-furanose-7-O-alpha-L-rhamnoside and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside or not according to the chromatographic information and mass spectrum information of the sample obtained in the step 3); if the composition contains kaempferol-3-O-beta-D-furanosyl-7-O-alpha-L-rhamnoside and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside, the composition is radix Hemsleyae Macrospermae; if not, the radix tetrastigme is the caulis sinomenii.
The kaempferol-3-O-beta-D-furanosyl-7-O-alpha-L-rhamnoside and the kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside are different compounds of the wisteria triloba and the sinomenium lyratum, and the wisteria triloba and the sinomenium triloba can be quickly and effectively distinguished by virtue of the two flavone characteristic compounds.
Specifically, the process of obtaining the differential compounds of the trefoil of wisteria floribunda and the trefoil of sinomenium graveolens in this example is as follows:
a UPLC-MS-MS method for rapidly screening the wisteria floribunda and the sinomenium cyrtonema, which comprises the following steps:
1) collecting the radix Stephaniae Tetrandrae and radix Stephaniae Tetrandrae to be distinguished, respectively preparing 20 batches of radix Stephaniae Tetrandrae and radix Stephaniae Tetrandrae of different production places into corresponding radix Stephaniae Tetrandrae and radix Stephaniae Tetrandrae test samples, pulverizing, sieving with 60 mesh sieve, and storing at-20 deg.C in dark place;
the information of the wisteria florbunda and the sinomenium tetrandra test sample is shown in the following table:
2) taking 1.0g of each of the test samples of the radix tetrastigme of the wisteria floribunda and the radix tetrastigme of the sinomenii in the step 1), precisely weighing, respectively placing in a conical bottle with a plug, precisely adding 10ml of 50% ethanol, weighing the weight, ultrasonically treating for 30 minutes, cooling, weighing the weight again, complementing the weight loss by using 50% ethanol, shaking up, centrifuging, precisely weighing 5ml of supernate, placing in a brown measuring flask of 25ml, adding 50% ethanol to the scale, shaking up, filtering, taking the subsequent filtrate, filtering to obtain a test sample solution, and then carrying out UPLC-MS-MS system analysis to respectively obtain the mass spectrum data corresponding to each of the radix tetrastigme of the wisteria floribunda and the test samples of the radix tetrastigme of the sinomenii floribunda;
wherein, the analysis conditions of the UPLC-MS-MS system are as follows:
chromatographic conditions are as follows: waters ACQUITY UPLC BEH C18 column, 2.1X 100mm, 1.7 μm; using 0.1% formic acid-water as mobile phase a and acetonitrile as mobile phase B, gradient elution is shown in the following table:
flow rate: 0.3 ml/min; the column temperature is 30 ℃; the detection wavelength is 254 nm;
mass spectrum conditions: the MS system is an ACQUITY UPLC-TQD-MS triple quadrupole mass spectrometer.
The conditions of use of the tandem mass spectrometer were: ionization mode ESI (-); the detection mode is as follows: detecting multiple reactions; capillary voltage: 3.0 Kv; taper hole voltage: 38V; the source temperature is 120 ℃; the temperature of the desolvation: 350 ℃; desolventizing flow rate: 500L/hr; taper hole air flow rate: 50L/hr; collision energy: 19 ev; residence time: 0.2 s; full scanning: the scan surface has a molecular weight of 100 to 1000.
3) Directly importing the mass spectrum data acquired in the step 2) into MassLynx V4.1 software, then carrying out data analysis, and carrying out chromatographic peak extraction and mass spectrum information characterization on each sample;
4) selecting a common compound chromatographic peak of two tetrastigma hemsleyanum Diels et Gilg as a reference, correcting chromatographic peak drift between samples by adopting a dynamic programming calculation method, aligning chromatographic peaks belonging to the same substance, registering the substance with closer retention time into a compound by utilizing a nearest clustering method, and finally establishing a registration chart of the sample and the chromatographic peak; after external standard correction, the method is used for subsequent analysis; as shown in fig. 1, which shows a high performance liquid chromatography comparison chart of a test sample of the wisteria floribunda and the sinomenium tetrandra, the result shows that the wisteria floribunda and the sinomenium tetrandra have obvious difference on an UPLC chromatogram.
5) Determining the difference compounds of the radix tetrastigme wisteria and the radix tetrastigme according to the peak area of the chromatogram and the mass number of the quasi-molecular ion peak corresponding to the peak-yielding retention time, directly introducing the original file of the mass spectrum information of the obtained difference substances into MassLynx V4.1 to further research the secondary mass spectrum information of the difference substances, and determining the substance structure, thereby obtaining the difference substances.
The main flavonoid compounds contained in the wisteria floribunda are shown in the following tables 1 and 2:
TABLE 1 relative peak area (mAU/g) of main flavonoids in wisteria trilobate
TABLE 2 main flavonoid component obtained by UPLC-MS/MS analysis of wisteria triloba
The main flavonoids in the tetrastigma hemsleyanum Diels et Gilg are shown in the following tables 3 and 4:
TABLE 3 relative peak area (mAU/g) of the major flavonoids in the Sinomenium acutum
TABLE 4 UPLC-MS/MS analysis of the main flavonoids from the vine Hemsley rockvine root
Fig. 2 shows a TIC comparison graph of the wisteria floribunda and sinomenium tetrandrum sample, total ion flow graph data of the wisteria floribunda and sinomenium tetrandrum sample is analyzed by Principal Component Analysis (PCA), and the result shows (as shown in fig. 5), each group of data is gathered in a respective 95% confidence limit ellipse graph, and the two groups of data can be completely separated, which indicates that the wisteria floribunda and sinomenium tetrafloribunda sample of two germplasms can be completely separated. Determining the differential compounds of the trefoil purpurea and the trefoil sinomenium according to the peak area of the chromatogram and the mass number of the quasi-molecular ion peak corresponding to the peak-yielding retention time, performing secondary mass spectrometry on the mass spectrum information of the main differential substances by using MassLynxV4.1 software, and determining the substance structure by combining reports of related compounds (refer to J.Nat.Med.,2011,65(2), 360-class 363, Chinese herbal medicines, 2017, 48(5), 874 and the like), thereby obtaining the main differential substances; the results show that:
1. kaempferol-3-O-beta-D-furanosyl-7-O-alpha-L-rhamnoside, kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside, kaempferol-3-O-beta-D-inumbioside, quercetin-7-O-alpha-L-rhamnoside, kaempferol-7-O-alpha-L-rhamnoside are main flavonoids in the samples of the Kadsura longipedunculata except common compound catechin (shown in figure 3 and table 2). Firework glycoside, quercetin-3-O-beta-D-xylosyl glucose-7-O-alpha-L-rhamnoside, robinin, isorhamnetin-3-O-beta-D-glucose-7-O-alpha-L-rhamnoside, rutin, isoquercitrin and astragalin in the radix tetrastigme leaf sample are main flavonoid components (figure 4 and table 4);
b. the high-content flavones in the tetrastigma hemsleyanum of the wisteria floribunda are kaempferol-3-O-beta-D-furanosyl-7-O-alpha-L-rhamnose and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside, but the tetrastigma hemsleyanum of the wisteria floribunda does not contain the flavones, so that the kaempferol-3-O-beta-D-furanosyl-7-O-alpha-L-rhamnose can be combined with the kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside to be used as the flavone characteristic compounds of the tetrastigma hemsleyanum.
In addition, the invention compares and determines the influence of different solvents and extraction modes on the total flavone content of the medicinal materials, the specific method is as follows, and the experimental results are shown in the following table.
1. Ultrasonic extraction method
Selecting radix Stephaniae Tetrandrae, removing impurities, pulverizing, sieving with No. 4 sieve, collecting 4 parts of 1.0g medicinal powder, adding 10ml of purified water, 50% ethanol, 70% ethanol and pure ethanol respectively, ultrasonic extracting for 30min, and filtering the extractive solution.
2. Reflux extraction method
Selecting radix Stephaniae Tetrandrae, removing impurities, pulverizing, sieving with No. 4 sieve, collecting 4 parts of 1.0g medicinal powder, adding 10ml of purified water, 50% ethanol, 70% ethanol and pure ethanol respectively, timing, heating in water bath, refluxing for 1 hr, taking out medicinal liquid, and filtering the extractive solution.
TABLE 5 results of different extraction modes and solvent effects on test sample parameters
The experimental result shows that the flavone content is related to the extraction mode and the solvent, but the extraction solvent is 50 percent and 70 percent of ethanol under the same extraction condition; the content of the flavone extracted by reflux is higher than that extracted by ultrasonic, but the UPLC-MS-MS analysis result shows that the proportion of the flavone in the ultrasonic extracting solution is obviously higher than that extracted by reflux, and the result can be that the flavonoid glycoside is unstable under the high-temperature condition or other impurities are more easily extracted under the high temperature condition. Therefore, in order to improve the proportion of the flavone in the radix tetrastigme test sample, the extraction process of the radix tetrastigme test sample is defined as 50 percent ethanol ultrasonic extraction for 30 min.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (6)
1. A method for rapidly detecting medicinal materials of wisteria floribunda and sinomenium acutum is characterized by comprising the following steps:
1) pulverizing radix tetrastigme medicinal materials to be distinguished, and sieving to prepare a sample to be tested;
2) taking the sample to be tested in the step 1), adding 10-20 times of 50-70% ethanol, carrying out ultrasonic extraction for 20-40 minutes to obtain a sample solution to be tested, and then carrying out UPLC-MS-MS system analysis to obtain mass spectrum data corresponding to the sample to be tested;
3) directly importing the data acquired in the step 2) into mass spectrometry software, then carrying out data analysis, and carrying out chromatographic peak extraction and mass spectrometry information characterization on a sample to be tested;
4) according to the chromatographic information and the mass spectrum information of the test sample obtained in the step 3), judging whether the test sample contains kaempferol-3-O-beta-D-furanapiose-7-O-alpha-L-rhamnoside and kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside for distinguishing the trefoil purpurea from the tetrastigma cochinchinensis through the peak area of the chromatogram, the mass number of an excimer ion peak corresponding to the peak-yielding retention time and the second-order mass spectrum information;
the chromatographic information and the mass spectrum information of the wisteria florbunda and the sinomenium tetrandra show that the wisteria florda and the sinomenium tetrandra can be completely separated according to the analysis result of the principal component analysis;
in the step 2), the analysis conditions of the UPLC-MS-MS system are as follows:
chromatographic conditions are as follows: waters ACQUITY UPLC BEH C18 column, 2.1X 100mm, 1.7 μm; using 0.1% formic acid-water as mobile phase a and acetonitrile as mobile phase B, gradient elution is shown in the following table:
Flow rate: 0.3 ml/min; the column temperature is 30 ℃; the detection wavelength is 254 nm;
mass spectrum conditions:
MS system ACQUITY UPLC-TQD-MS triple quadrupole mass spectrometer; the conditions of use of the tandem mass spectrometer were: an ionization mode, namely an ESI negative ion detection mode; the detection mode is as follows: detecting multiple reactions; capillary voltage: 3.0 Kv; taper hole voltage: 38V; the source temperature is 120 ℃; the temperature of the desolvation: 350 ℃; desolventizing flow rate: 500L/hr; taper hole air flow rate: 50L/hr; collision energy: 19 ev; residence time: 0.2 s; full scanning: the scan surface has a molecular weight of 100 to 1000.
2. The method for rapidly detecting the medicinal materials of the Chinese wisteria roots and the Chinese ivy leaves as claimed in claim 1, wherein the kaempferol-3-O-beta-D-furanapiose-7-O-alpha-L-rhamnoside in the step 4): molecular weight of 726, molecular formula C32H38O19The secondary mass spectral fragments are 647.5,527.4,486.6,431.9,392.1, 289.7.
3. The method of claim 1, wherein the method comprises the step of rapidly detecting the trefoil of wisteria floribundaThe method for mixing with the medicinal material of the caulis sinomenii and radix tetrastigme is characterized in that the kaempferol-7-O-alpha-L-rhamnose-3-O-beta-D-glucopyranoside in the step 4): molecular weight is 594, molecular formula C27H30O15The secondary mass spectral fragments were 515.5,451.7,386.5,291.0,246.5.
4. The method for rapidly detecting the medicinal materials of the Chinese wistaria and the Chinese wistaria in the claim 1, wherein in the step 1), the sample to be tested is pulverized and sieved by a 60-mesh sieve, and is stored in the dark at the temperature of minus 20 ℃.
5. The method for rapidly detecting the medicinal materials of the wisteria floribunda and the sinomenium acutum DC according to the claim 1, which is characterized in that in the step 2), 1.0g of the sample to be detected in the step 1) is taken and precisely weighed, the sample is placed in a conical flask with a stopper, 10ml of 50% ethanol is precisely added, the weight is weighed, the ultrasonic treatment is carried out for 30 minutes, the sample is cooled, the weight is weighed again, the 50% ethanol is used for complementing the weight loss, the sample is shaken up and evenly centrifuged, 5ml of supernatant is precisely measured and placed in a brown measuring flask with 25ml, the 50% ethanol is added to the scale, the shaking up and even, the filtration is carried out, and the subsequent filtrate is filtered, so as to obtain the sample solution.
6. The method for rapidly detecting the medicinal materials of the Chinese wisteria roots and the Chinese wisteria roots according to claim 1, wherein in the step 3), the mass spectrometry software is MassLynx V4.1 software.
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