CN114441680B - Method for distinguishing traditional Chinese medicine fructus aurantii from garden incense based on high-resolution mass spectrometry technology - Google Patents
Method for distinguishing traditional Chinese medicine fructus aurantii from garden incense based on high-resolution mass spectrometry technology Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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Abstract
The invention discloses a method for distinguishing traditional Chinese medicine fructus aurantii from a garden incense based on a high-resolution mass spectrometry technology. The invention is mainly used for rapidly identifying the traditional Chinese medicine fructus aurantii and the garden incense. The invention develops a method for distinguishing traditional Chinese medicine fructus aurantii from a garden based on an ultra-high performance liquid quadrupole time-of-flight mass spectrometry technology, establishes a special distinguishing method of traditional Chinese medicine-characteristic components, finds a characteristic compound-hydrated oxidized peucedanum element capable of distinguishing fructus aurantii from a garden, and can distinguish the fructus aurantii from the garden quickly and effectively; the invention adopts the conventional morphological appearance-based experience identification method different from the bitter orange and the aromatic garden, avoids subjective fault tolerance, enhances the specificity of the method, can effectively solve the difficult problem that the traditional Chinese medicine bitter orange and the aromatic garden are difficult to distinguish in actual work, and provides a technical method and basis for striking illegal behaviors of the traditional Chinese medicine bitter orange which is impersonated by the mixed and counterfeited product of the aromatic garden.
Description
Technical Field
The invention relates to the technical field of medicine analysis and detection, in particular to a method for distinguishing traditional Chinese medicine fructus aurantii from garden incense based on a high-resolution mass spectrum technology.
Background
Fructus Aurantii is dried immature fruit of Citrus aurantium L and its cultivar. The garden is a dry mature fruit of a rue plant garden Citrus wilsonii Tanaka, which is one of sources of traditional Chinese medicine citron. The dried immature fruits in the incense garden have the appearance very similar to that of the bitter orange, and the consumption and the price of the bitter orange are higher than those of the incense circle, so that the incense circle is often sold by illegal drug vendors as the bitter orange after being impossed, and the conventional experience identification based on the morphological appearance is difficult to distinguish and identify the conditions. In addition, due to the lack of professional identifiers of traditional Chinese medicines, the authenticity of the raw material medicines of the fructus aurantii is difficult to control for Chinese patent medicine production enterprises, and the integrity of the raw material medicines is destroyed by the finished product of the traditional Chinese patent medicines, so that the difficulty of identifying and distinguishing the traditional Chinese medicine inspection personnel is increased to a certain extent. In clinical practice, fructus Aurantii has the effects of regulating qi, relieving middle energizer, promoting qi stagnation and relieving distention, and is commonly used for treating chest and hypochondrium qi stagnation, distention and pain, indigestion, phlegm-fluid retention, visceral organ prolapse and the like. The garden has the effects of soothing liver, regulating qi, relieving the middle warmer and reducing phlegm, and is commonly used for treating liver and stomach qi stagnation, chest and hypochondrium distending pain, epigastric fullness, vomiting, hiccup, excessive phlegm, cough and other diseases. The former is good at relieving distension, the latter is good at soothing liver and resolving phlegm, and the main effects of the two are obviously different, so that the clinical efficacy is seriously affected by mixed filling.
At present, the identification means of the mixed and fake traditional Chinese medicine mainly comprise methods such as thin layer chromatography identification, microscopic identification, traditional Chinese medicine characteristic spectrum or fingerprint spectrum identification, polymerase Chain Reaction (PCR) and the like, but most of the means are the identification of the true and false of the traditional Chinese medicine raw materials, and the condition of false negative easily occurs to the varieties of the finished medicines or similar varieties, so that the identification methods are not suitable for the identification of the bitter orange and the garden.
Therefore, it is important to find a method with high efficiency and strong specificity for identifying traditional Chinese medicine fructus Aurantii and garden incense.
Disclosure of Invention
The invention aims to overcome the technical defects of the background technology and provides a method for distinguishing traditional Chinese medicine fructus aurantii from a garden incense based on a high-resolution mass spectrum technology. The invention is mainly used for rapidly identifying the traditional Chinese medicine fructus aurantii and the garden incense. The invention is mainly used for a supplementary identification method which is based on the conventional morphological appearance identification and can not be further distinguished when the traditional Chinese medicine is determined to be the fructus aurantii or the garden incense. Compared with the conventional morphological appearance identification, the method adopts a high-resolution mass spectrometry technology, and can effectively distinguish the bitter orange from the garden under the condition that the conventional morphological appearance identification is not easy to distinguish or the complete morphological appearance is lost.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method for distinguishing traditional Chinese medicine fructus Aurantii from garden incense based on high resolution mass spectrum technology comprises the following steps:
s1, collecting representative fructus aurantii, garden balsam and pomelo medicinal materials, respectively extracting by adopting alcohol reagents to obtain respective sample solutions, and acquiring a total ion flow diagram of the obtained sample solutions by adopting ultra-high performance liquid chromatography and quadrupole time-of-flight mass spectrometry;
s2, screening characteristic components which can be distinguished from each other from a total ion flow diagram of three medicinal materials, namely, the collected fructus aurantii, the garden balsam and the pomelo, so as to obtain the mass number of the corresponding characteristic components;
s3, identifying the components of the compound according to the quality number of the obtained characteristic components and the corresponding primary and secondary mass spectrograms through literature and library searching technology, and obtaining the compound;
s4, establishing a method for distinguishing traditional Chinese medicine fructus aurantii from a garden according to known characteristic components and extraction conditions of a sample to be detected and high-resolution mass spectrum conditions;
s5, the sample to be detected adopts the established method of the S4, simultaneously, the characteristic compound reference substance and the mass spectrogram of the sample to be detected are collected, the ion flow diagram of the characteristic compound is extracted, and the same molecular ion peak is presented at the retention time consistent with the ion flow diagram of the characteristic compound reference substance.
Preferably, in the above method, in S1, the representative samples of fructus Aurantii, fructus Citri Grandis and fructus Citri Grandis are collected in the main production area of the country, and the coverage area includes Jiangxi, zhejian, fujian, hunan, etc. with a certain regional representativeness. Fructus Aurantii identified as Citrus aurantium L. Of Rutaceae, citrus paradisi Citrus wilsonii Tanaka, and Rutaceae, citrus maxima (Burm) Merr.
Preferably, in the above method, in S1, the alcohol reagent is methanol (analytically pure); preferably, the extraction is carried out by using an ultrasonic extraction method for 30min, wherein the addition amount of the methanol is 25ml calculated by 0.1g of traditional Chinese medicinal materials.
Preferably, in the method, the condition of the ultrasonic wave is 250W and 40kHz.
Preferably, in the above method, in S1, the total ion flow diagram of fructus Aurantii, fructus Citri Grandis and its molecular ion peak extracted at m/z305.1021 are shown in FIG. 1.
Preferably, in the above method, in S2, the screening process of the characteristic components is to collect MS/MS mass spectrum data by adopting a UPLC-Q-TOF high-resolution mass spectrometry, introduce Agilent MPP software for PCA and multivariate discriminant analysis, establish a discriminant model of fructus Aurantii, xiang yuan, and fructus Citri grandis, and screen the differential components among groups; the total number of the unique components of the incense circles is 53 (see figure 2); the method comprises the steps of searching and matching by adopting a compound database of Agilent TCM library-V20-04-17, selecting 9 compounds identified by matching of data parameters of a database search mass spectrum, screening (table 1), and verifying by combining a comparison quality spectrogram to find out hydrated peucedanin Oxypeucedanin hydrate (mass spectrum information of the hydrated peucedanin is shown in table 2, a total ion flow diagram of the hydrated peucedanin and molecular ion peaks extracted at m/z305.1021 are shown in figure 1, and a typical mass spectrogram of the hydrated peucedanin is shown in figure 3), and can effectively distinguish a fragrance garden from a bitter orange to be a fragrance circle specific component, so that a mass spectrum screening method is established.
In the technical scheme, the purpose of adopting the pomelo is to reduce noise of cluster analysis, reduce the screening range of the characteristic components of the garden in the discrimination model, and facilitate the simplification of experimental flow.
TABLE 1 search and identification Table of the library of spectrum of differential compounds of the invention
Preferably, in the above method, in S1, the ultra-high performance liquid chromatography uses the following conditions:
chromatographic conditions: chromatographic column: an Agilent SB C18 (100 mm. Times.2.1 mm,1.8 μm) column; column temperature: 30 ℃; flow rate: 0.4ml/min; binary gradient elution is adopted for detection, the mobile phase A is acetonitrile, and the mobile phase B is 0.1% formic acid aqueous solution; from 0 to 7 minutes, based on 100% of the total volume of the mobile phase, the volume of the mobile phase A is increased from 15% to 25%, and the volume of the mobile phase B is decreased from 85% to 75%; at 7-8 min, the volume of the mobile phase A is increased from 25% to 40%, and the volume of the mobile phase B is decreased from 75% to 60%; at 8-10 min, the volume of the mobile phase A is increased from 40% to 45%, and the volume of the mobile phase B is decreased from 60% to 55%; at 10-13 min, the volume of the mobile phase A is increased from 45% to 60%, and the volume of the mobile phase B is decreased from 55% to 40%; at 13-15 min, the volume of the mobile phase A is reduced from 60% to 15%, and the volume of the mobile phase B is increased from 40% to 85%. .
Preferably, in the method, in S1, the quadrupole time-of-flight mass spectrum uses the following conditions:
ion source: electrospray ionization (ESI); positive ion scan mode: electrospray voltage: 5500V; ion source temperature: 550 ℃; air curtain gas: 35psi; atomizing gas: 55psi; auxiliary gas: 55psi; cluster removal voltage: 60V; collision voltage: 35V; the scanning mode is primary mass spectrum full scanning and secondary mass spectrum full scanning, and the scanning range is 100-1500 m/z; reference ion m/z:121.0480 and 922.0098.
Preferably, in the method, in step S3, searching and matching the sample retention time, the primary accurate mass number and the secondary mass spectrogram with a mass spectrum software traditional Chinese medicine database to identify the characteristic compound;
preferably, in the above method, in S3, the characteristic compound is a hydrated oxidized peucedanum praeruptorum (Oxypeucedanin hydrate), which is a characteristic component of a garden incense, and has a molecular weight of 304.29Da; MS (MS) 1 (m/z)305.1021[M+H] + ;MS 2 (m/z)203.0345[M+H] + 、147.0437[M+H] + The method comprises the steps of carrying out a first treatment on the surface of the A typical mass spectrum of the hydrated oxidized peucedanum praeruptorum Oxypeucedanin hydrate is shown in FIG. 3.
Preferably, in the above method, in S4, the method for identifying the genuine product of fructus Aurantii is as follows: the ion flow chromatograph of the sample to be extracted with the mass-to-charge ratio (m/z) 305.1021 should not show a chromatographic peak corresponding to the reference substance, and if the corresponding chromatographic peak is shown, the corresponding secondary mass spectrum should not be consistent with the reference substance.
TABLE 2 mass spectrum information table of the hydrated oxidized peucedanum praeruptorin Oxypeucedanin hydrate of the invention
A method for distinguishing traditional Chinese medicine fructus Aurantii from garden incense based on high resolution mass spectrum technology comprises the following steps:
(1) Preparing a reference substance solution: taking a proper amount of characteristic components of the hydrated oxidized peucedanum praeruptorum, precisely weighing, adding an alcohol reagent for dissolution, and obtaining a reference substance solution;
(2) Sample solution preparation: taking a sample medicinal material, precisely weighing, and extracting with an alcohol reagent to obtain a sample solution;
(3) And (3) measuring: respectively collecting total ion flow diagrams of the reference substance solution and the sample solution by adopting ultra-high performance liquid chromatography and quadrupole time-of-flight mass spectrometry, and respectively extracting a primary mass spectrogram and/or a secondary mass spectrogram of the total ion flow diagrams;
(4) And (3) result judgment: if the first-order mass spectrum of the sample solution extracted by the mass-to-charge ratio (m/z) 305.1021 does not have chromatographic peaks corresponding to the first-order mass spectrum of the reference solution extracted by the same mass-to-charge ratio, the sample is a fructus aurantii genuine product;
conversely, if the first-order mass spectrum of the sample solution extracted at the mass-to-charge ratio (m/z) 305.1021 shows a chromatographic peak corresponding to the first-order mass spectrum of the reference solution extracted at the same mass-to-charge ratio, if the second-order mass spectrum of the sample solution extracted at the mass-to-charge ratios (m/z) 203.0345 and 147.0437 does not show a chromatographic peak corresponding to the second-order mass spectrum of the reference solution extracted at the same mass-to-charge ratio, the sample is a fructus Aurantii genuine product;
conversely, if the secondary mass spectrum of the sample solution extracted at mass-to-charge ratios (m/z) 203.0345 and 147.0437 shows chromatographic peaks respectively corresponding to the secondary mass spectrum of the reference solution extracted at the same mass-to-charge ratio, the sample is in a garden adulteration.
In the above technical scheme, in the step (2), the sample to be identified is a suspected sample of fructus Aurantii or garden which is indistinguishable based on conventional morphological appearance identification.
The application of the hydrated oxidized peucedanin in distinguishing traditional Chinese medicine fructus aurantii from garden incense based on high-resolution mass spectrum technology.
The basic principle of the invention is as follows:
with the rapid development of mass spectrometry technology, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) has the advantages of high analysis speed, high sensitivity and resolution, simultaneous determination of multiple components, and obvious advantages in separation and analysis of endogenous components in near-source traditional Chinese medicines and complex compound patent medicine matrixes.
According to the situation, a special identification method of traditional Chinese medicine-characteristic components is established, a method for distinguishing traditional Chinese medicine fructus aurantii from a fragrance garden based on an ultra-high performance liquid quadrupole time-of-flight mass spectrometry technology is developed, the problem that the traditional Chinese medicine fructus aurantii and the fragrance garden are difficult to distinguish in actual work can be effectively solved, and a technical method and a basis are provided for striking illegal behaviors that a fragrance garden mixed counterfeit product impersonates the traditional Chinese medicine fructus aurantii.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention develops a method for distinguishing traditional Chinese medicine fructus aurantii from a garden based on an ultra-high performance liquid quadrupole time-of-flight mass spectrometry technology, establishes a special distinguishing method of traditional Chinese medicine-characteristic components, finds out a characteristic compound which can distinguish fructus aurantii from a garden, namely hydrated oxidized peucedanum praeruptorum Oxypeucedanin hydrate, and can distinguish the fructus aurantii from the garden quickly and effectively;
(2) The invention adopts the conventional morphological appearance-based experience identification method different from the bitter orange and the aromatic garden, avoids subjective fault tolerance, enhances the specificity of the method, can effectively solve the difficult problem that the traditional Chinese medicine bitter orange and the aromatic garden are difficult to distinguish in actual work, and provides a technical method and basis for striking illegal behaviors of the traditional Chinese medicine bitter orange which is impersonated by the mixed and counterfeited product of the aromatic garden.
Drawings
FIG. 1 is a total ion flow diagram of fructus Aurantii, fructus Citri Grandis, and hydrated peucedanum oxide of the present invention (A. Fructus Aurantii total ion flow diagram; C. Fructus Citri Grandis total ion flow diagram; E. Fructus Citri Grandis total ion flow diagram; G. Pre-hydrated oxidation Hu Suzong ion flow diagram) and molecular ion peaks extracted at m/z305.1021 (B. Fructus Aurantii molecular ion peak; D. Fructus Citri Grandis molecular ion peak; F. Fructus Citri Grandis molecular ion peak; H. Hydrated oxidation peucedanum oxide molecular ion peak);
FIG. 2 is a diagram of the wien's differential compounds of fructus Aurantii, xiang yuan and fructus Citri Grandis of the present invention;
FIG. 3 is a typical mass spectrum of the hydrated oxidized peucedanin Oxypeucedanin hydrate of the present invention;
FIG. 4 shows the mass spectrum screening results of the Chinese patent medicine preparation in Baoxinning of the embodiment 1 (A. The total ion flow chart of the test sample; B. The first mass spectrum of the test sample (m/z 305.1021), C. The second mass spectrum of the test sample (m/z 203.0345), D. The second mass spectrum of the test sample (m/z 147.0437), E. The total ion flow chart of the negative sample; F. The first mass spectrum of the negative sample (m/z 305.1021), G. The second mass spectrum of the negative sample (m/z 203.0345), H. The second mass spectrum of the negative sample (m/z 147.0437), I. The total ion flow chart of the reference sample; J. The first mass spectrum of the reference sample (m/z 305.1021), K. The second mass spectrum of the reference sample (m/z 203.0345), L. The second mass spectrum of the reference sample (m/z 147.0437), wherein the test sample is the Chinese patent medicine preparation to be tested, the negative sample is the Chinese patent medicine preparation of the fructus Aurantii positive charge and the Chinese patent medicine preparation, and the negative sample is the hydrated as the oxidation product
Detailed Description
For a better understanding of the present invention, reference will be made to the following description of specific embodiments and accompanying drawings. It is to be understood that these examples are provided only for further illustration of the present invention and are not intended to limit the scope of the present invention. It should be further understood that after reading the description of the present invention, those skilled in the art make some insubstantial changes or modifications to the present invention, which still fall within the scope of the present invention.
Example 1
Identification of the pseudo-product of the garden in the Baoxinning traditional Chinese medicine preparation (early identification already determines that the formula in the Baoxinning traditional Chinese medicine preparation contains the bitter orange or the garden but cannot be further distinguished):
1.1 chromatographic conditions
Chromatographic column: an Agilent SB C18 (100 mm. Times.2.1 mm,1.8 μm) column; column temperature: 30 ℃; flow rate: 0.4ml/min; binary gradient elution (acetonitrile as mobile phase a, 0.1% formic acid in water as mobile phase B) was used: 0-7 min,15% -25% A,85% -75% B; 7-8 min,25% -40% A,75% -60% B; 8-10 min,40% -45% A,60% -55% B; 10-13 min,45% -60% A,55% -40% B; 13-15 min,60% -15% A,40% -85% B.
1.2 Mass Spectrometry conditions
Ion source: electrospray ionization (ESI); positive ion scan mode: electrospray voltage: 5500V; ion source temperature: 550 ℃; air curtain gas: 35psi; atomizing gas: 55psi; auxiliary gas: 55psi; cluster removal voltage: 60V; collision voltage: 35V; the scanning modes are primary mass spectrum full scanning (MS) and secondary mass spectrum full scanning (MS/MS), and the scanning ranges are 100-1500 m/z; reference ion m/z:121.0480 and 922.0098.
1.3 preparation of control solution
Taking a proper amount of a hydrated oxidized peucedanum praeruptorum reference substance, precisely weighing, adding methanol to prepare a solution containing 2 mug per 1ml, and obtaining the reference substance solution.
1.4 preparation of negative sample solutions
Taking a negative sample, namely about 0.1g of the finished product of the fructus aurantii, adding the materials and the finished product of the Baoxinning, precisely weighing, placing into a conical bottle with a plug, precisely adding 25ml of methanol, weighing, performing ultrasonic treatment (250W, 40 kHz) for 30min, cooling, weighing again, supplementing the lost weight with methanol, shaking uniformly, filtering, and taking a subsequent filtrate to obtain a negative sample solution.
1.5 preparation of sample solutions
Taking about 0.1g of a test sample, namely the Chinese patent medicine powder of the Baoxinning to be tested, precisely weighing, placing into a conical bottle with a plug, precisely adding 25ml of methanol, weighing, performing ultrasonic treatment (250W, 40 kHz) for 30min, cooling, weighing again, supplementing the lost weight with methanol, shaking uniformly, filtering, and taking a subsequent filtrate to obtain a test sample solution.
1.6 assay
2 mu l of the reference substance solution, 1-2 mu l of the negative sample solution and 1-2 mu l of the test substance solution are respectively sucked precisely, and are injected into a liquid chromatograph-mass spectrometer (Agilent 1290-UPLC and Agilent 6545Q-TOF) for measurement, and a primary mass spectrogram and a secondary mass spectrogram are recorded.
1.7 result determination
If the first-order mass spectrum of the sample solution extracted by the mass-to-charge ratio (m/z) 305.1021 does not have a chromatographic peak corresponding to the first-order mass spectrum of the reference sample solution extracted by the same mass-to-charge ratio, the sample is not mixed with the material of the incense garden and is a qualified medicine, namely the sample is a finished product of the fructus aurantii, the material of the sample is added and the heart is protected;
on the contrary, if the first-order mass spectrum of the sample solution extracted with the mass-to-charge ratio (m/z) 305.1021 shows a chromatographic peak corresponding to the first-order mass spectrum of the reference solution extracted with the same mass-to-charge ratio, since different varieties of fructus Aurantii may have other components (i.e., isomers) with the same molecular weight as those of the hydrated oxidized peucedanin, in order to eliminate the interference of the matrix, the second-order mass spectrum is needed to be further used for judgment;
further, if the secondary mass spectrograms of the sample solution extracted with the mass-to-charge ratios (m/z) 203.0345 and 147.0437 do not have chromatographic peaks respectively corresponding to the secondary mass spectrograms of the reference sample solution extracted with the same mass-to-charge ratios, the sample is not mixed with the material of the artificial incense garden and is a qualified medicine, namely the sample is a finished product of the fructus aurantii, namely the material of the sample is added to the Chinese patent medicine of the Baoxinning;
otherwise, if chromatographic peaks respectively corresponding to the secondary mass spectrograms of the sample solution extracted with the mass-to-charge ratios (m/z) 203.0345 and 147.0437 appear in the secondary mass spectrograms of the reference sample solution extracted with the same mass-to-charge ratio, the sample is fed into the pseudo-aromatic garden and is a disqualified drug, namely the sample is a finished drug preparation in the bitter orange pseudo-aromatic garden feeding and heart protecting state.
As can be seen from F and J in fig. 4, since the first-order mass spectrum of the negative sample extracted with the mass-to-charge ratio (m/z) 305.1021 does not show a chromatographic peak corresponding to the first-order mass spectrum of the reference substance extracted with the mass-to-charge ratio (m/z) 305.1021, the negative sample does not contain the hydrated oxidized peucedanum praeruptorum, thereby indicating that the negative sample is not doped with the artificial incense garden charge and is a qualified medicine, namely the negative sample is the finished product of fructus aurantii, the material is added and the medicine is prepared in the Baoxinning.
As can be seen from B, C, D and J, K, L in fig. 4, since the first-order mass spectrum of the sample solution extracted with the mass-to-charge ratio (m/z) 305.1021 shows a chromatographic peak corresponding to the first-order mass spectrum of the reference hydrated peucedanum oxide extracted with the mass-to-charge ratio (m/z) 305.1021, and the second-order mass spectrum of the sample solution extracted with the mass-to-charge ratios (m/z) 203.0345 and 147.0437 shows a chromatographic peak corresponding to the second-order mass spectrum of the reference hydrated peucedanum oxide extracted with the mass-to-charge ratios (m/z) 203.0345 and 147.0437, respectively, it is indicated that the sample contains the hydrated peucedanum oxide, thereby indicating that the sample is a adulterated incense garden charge, i.e., the sample is a finished drug in the bitter orange adulteration incense garden charge and the heart protection.
The above description is not intended to limit the invention, nor is the invention limited to the examples described above. Variations, modifications, additions, or substitutions will occur to those skilled in the art and are therefore within the spirit and scope of the invention.
Claims (1)
1. The method for distinguishing the traditional Chinese medicine fructus aurantii from the garden incense based on the high-resolution mass spectrum technology is characterized by comprising the following steps of:
(1) Preparing a reference substance solution: taking a proper amount of characteristic components of the hydrated oxidized peucedanum praeruptorum, precisely weighing, adding an alcohol reagent for dissolution, and obtaining a reference substance solution;
(2) Sample solution preparation: taking a medicinal material of a test sample, precisely weighing, adding an alcohol reagent for dissolution to obtain a solution of the test sample;
(3) And (3) measuring: collecting total ion flow diagrams of the reference substance solution and the sample solution respectively by adopting ultra-high performance liquid chromatography and quadrupole time-of-flight mass spectrometry, and extracting a primary mass spectrogram of the total ion flow diagrams or a primary mass spectrogram and a secondary mass spectrogram of the total ion flow diagrams respectively;
(4) And (3) result judgment: if the first-order mass spectrum of the sample solution extracted with the mass-to-charge ratio m/z305.1021 does not have a chromatographic peak corresponding to the first-order mass spectrum of the reference sample solution extracted with the same mass-to-charge ratio, the sample is a fructus aurantii genuine product;
conversely, if the first-order mass spectrum of the sample solution extracted with the mass-to-charge ratio m/z305.1021 shows a chromatographic peak corresponding to the first-order mass spectrum of the reference sample solution extracted with the same mass-to-charge ratio, if the second-order mass spectrum of the sample solution extracted with the mass-to-charge ratios m/z 203.0345 and 147.0437 does not show a chromatographic peak corresponding to the second-order mass spectrum of the reference sample solution extracted with the same mass-to-charge ratio, the sample is a fructus aurantii genuine product;
otherwise, if chromatographic peaks respectively corresponding to the secondary mass spectrograms of the sample solution extracted with the mass-to-charge ratios m/z 203.0345 and 147.0437 appear in the secondary mass spectrograms of the reference sample solution extracted with the same mass-to-charge ratios, the sample is in a fragrance garden adulteration;
in the step (1), the screening process of the characteristic components adopts a UPLC-Q-TOF high-resolution mass spectrometry to collect MS/MS mass spectrum data, leads Agilent MPP software to carry out PCA and multivariate discriminant analysis, establishes a discriminant model of fructus Aurantii, xiang yuan and shaddock, and screens the differential components among groups; obtaining 53 unique components of the sweet circles; selecting 9 compounds which are identified by matching the mass spectrum data parameters retrieved by a database, screening, and combining with a comparison quality spectrogram to verify that the hydrated oxypeucedanin can effectively distinguish the incense circle from the fructus aurantii and is a peculiar component of the incense circle, thereby establishing a mass spectrum screening method;
in the step (1), the alcohol reagent is methanol; adding the methanol to prepare a solution with 2 mug per 1ml, thus obtaining the reference substance solution;
in the step (2), the alcohol reagent is methanol; when in dissolution, the adding amount of the methanol is 25ml calculated by 0.1g of Chinese medicinal materials, and ultrasonic treatment is carried out for 30min;
in the step (3), respectively precisely sucking the reference substance solution 2 [ mu ] l and the sample solution 1-2 [ mu ] l, injecting into a liquid chromatograph-mass spectrometer for measurement, and recording a primary mass spectrogram or respectively recording a primary mass spectrogram and a secondary mass spectrogram;
in the step (3), the ultra performance liquid chromatography uses the following conditions:
chromatographic conditions: chromatographic column: an Agilent SB C18 chromatographic column, 100mm x 2.1mm,1.8 μm; column temperature: 30 ℃; flow rate: 0.4ml/min; binary gradient elution is adopted, the mobile phase A is acetonitrile, and the mobile phase B is 0.1% formic acid aqueous solution; at 0-7 min, the volume of the mobile phase A is increased from 15% to 25% and the volume of the mobile phase B is decreased from 85% to 75% by taking the total volume of the mobile phase as 100%; at 7-8 min, the volume of the mobile phase A is increased from 25% to 40%, and the volume of the mobile phase B is decreased from 75% to 60%; at 8-10 min, the volume of the mobile phase A is increased from 40% to 45%, and the volume of the mobile phase B is decreased from 60% to 55%; at 10-13 min, the volume of the mobile phase A is increased from 45% to 60%, and the volume of the mobile phase B is decreased from 55% to 40%; at 13-15 min, the volume of the mobile phase A is reduced from 60% to 15%, and the volume of the mobile phase B is increased from 40% to 85%;
in the step (3), the quadrupole time-of-flight mass spectrum uses the following conditions:
ion source: electrospray ionization ESI; positive ion scan mode: electrospray voltage: 5500V; ion source temperature: 550 ℃; air curtain gas: 35psi; atomizing gas: 55psi; auxiliary gas: 55psi; cluster removal voltage: 60V; collision voltage: 35V; the scanning mode is primary mass spectrum full scanning and secondary mass spectrum full scanning, and the scanning range is 100-1500 m/z; reference ion m/z:121.0480 and 922.0098.
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Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103954724A (en) * | 2014-04-28 | 2014-07-30 | 四川逢春制药有限公司 | Method for detecting Jingfang granules |
CN107389813A (en) * | 2017-07-10 | 2017-11-24 | 北京中医药大学 | Rascal, dried orange peel, the dried immature fruit of citron orange and the method for Fructus Aurantii are differentiated based on chemical classification and UPLC Tof MS |
CN109828052A (en) * | 2019-03-13 | 2019-05-31 | 西南民族大学 | A kind of quick identification Rhizoma Et Radix Notopterygii type, the true and false or whether there is or not method |
CN110530999A (en) * | 2019-09-18 | 2019-12-03 | 山东省食品药品检验研究院 | A kind of distinguishing method between true and false of caribou horn |
CN110715994A (en) * | 2019-09-30 | 2020-01-21 | 山西大学 | Method for analyzing difference chemical components of spina date seed and spina date seed by using UHPLC-Q-Orbitrap MS |
CN111198236A (en) * | 2020-01-14 | 2020-05-26 | 山东中医药大学 | Method for identifying honeysuckle and lonicera confusa by utilizing high performance liquid chromatography and liquid chromatography-mass spectrometry |
KR102124999B1 (en) * | 2019-12-23 | 2020-06-22 | 조선대학교산학협력단 | A method for distinguishing between Angelica decursiva and Anthriscus sylvestris |
CN112114073A (en) * | 2020-10-24 | 2020-12-22 | 中国科学院昆明植物研究所 | Fructus aurantii quality marker, screening method and application thereof |
CN112557563A (en) * | 2019-09-10 | 2021-03-26 | 泰州医药城国科化物生物医药科技有限公司 | Method for identifying medicinal material fructus aurantii in western and Jiangxi regions |
CN112903890A (en) * | 2021-02-02 | 2021-06-04 | 湖北省食品质量安全监督检验研究院 | Method for identifying traditional Chinese medicine components in food based on high-resolution mass spectrometry technology |
CN113030325A (en) * | 2021-03-11 | 2021-06-25 | 广东一方制药有限公司 | Method for constructing and identifying characteristic maps of fructus aurantii and fructus aurantii stir-fried with bran |
CN113030332A (en) * | 2021-03-10 | 2021-06-25 | 谱尼测试集团北京检验认证科学研究院有限公司 | Method for rapidly identifying authenticity of rhizoma atractylodis bulk drug |
CN113759003A (en) * | 2020-06-04 | 2021-12-07 | 合肥卫元堂中药股份有限公司 | Licorice origin distinguishing method based on UPLC fingerprint spectrum and chemometrics method |
-
2022
- 2022-01-26 CN CN202210095206.1A patent/CN114441680B/en active Active
Patent Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103954724A (en) * | 2014-04-28 | 2014-07-30 | 四川逢春制药有限公司 | Method for detecting Jingfang granules |
CN107389813A (en) * | 2017-07-10 | 2017-11-24 | 北京中医药大学 | Rascal, dried orange peel, the dried immature fruit of citron orange and the method for Fructus Aurantii are differentiated based on chemical classification and UPLC Tof MS |
CN109828052A (en) * | 2019-03-13 | 2019-05-31 | 西南民族大学 | A kind of quick identification Rhizoma Et Radix Notopterygii type, the true and false or whether there is or not method |
CN112557563A (en) * | 2019-09-10 | 2021-03-26 | 泰州医药城国科化物生物医药科技有限公司 | Method for identifying medicinal material fructus aurantii in western and Jiangxi regions |
CN110530999A (en) * | 2019-09-18 | 2019-12-03 | 山东省食品药品检验研究院 | A kind of distinguishing method between true and false of caribou horn |
CN110715994A (en) * | 2019-09-30 | 2020-01-21 | 山西大学 | Method for analyzing difference chemical components of spina date seed and spina date seed by using UHPLC-Q-Orbitrap MS |
KR102124999B1 (en) * | 2019-12-23 | 2020-06-22 | 조선대학교산학협력단 | A method for distinguishing between Angelica decursiva and Anthriscus sylvestris |
CN111198236A (en) * | 2020-01-14 | 2020-05-26 | 山东中医药大学 | Method for identifying honeysuckle and lonicera confusa by utilizing high performance liquid chromatography and liquid chromatography-mass spectrometry |
CN113759003A (en) * | 2020-06-04 | 2021-12-07 | 合肥卫元堂中药股份有限公司 | Licorice origin distinguishing method based on UPLC fingerprint spectrum and chemometrics method |
CN112114073A (en) * | 2020-10-24 | 2020-12-22 | 中国科学院昆明植物研究所 | Fructus aurantii quality marker, screening method and application thereof |
CN112903890A (en) * | 2021-02-02 | 2021-06-04 | 湖北省食品质量安全监督检验研究院 | Method for identifying traditional Chinese medicine components in food based on high-resolution mass spectrometry technology |
CN113030332A (en) * | 2021-03-10 | 2021-06-25 | 谱尼测试集团北京检验认证科学研究院有限公司 | Method for rapidly identifying authenticity of rhizoma atractylodis bulk drug |
CN113030325A (en) * | 2021-03-11 | 2021-06-25 | 广东一方制药有限公司 | Method for constructing and identifying characteristic maps of fructus aurantii and fructus aurantii stir-fried with bran |
Non-Patent Citations (3)
Title |
---|
基于UPLC-QTOF-MS/MS 分析酸橙果实中的生物活性成分;秦艳;《食品与发酵工业》;全文 * |
枳壳与香圆枳壳的鉴别;沈廷明;《StraitPharmaceuticalJournal》;全文 * |
正品枳壳指纹图谱的构建及其与混伪品的聚类分析;庞文静;《中南药学》;全文 * |
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