CN114113444A - Method for identifying traditional Chinese medicine derived from Panax plant or different parts thereof and application thereof - Google Patents

Abstract

The invention relates to the technical field of traditional Chinese medicine identification, in particular to an identification method of traditional Chinese medicines which are derived from various plants of panax or different parts of panax and have similar chemical compositions and application thereof. The invention uses notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, malonyl ginsenoside Re1, 24(R) -pseudoginsenoside F11 and the like as markers to detect a sample to be detected, and identifies the traditional Chinese medicine source of the component to be detected according to the detection result. The content of the above marked components in the traditional Chinese medicines of different Panax plants and different parts of the same Panax plant is different, the specific variety and the part of the sample to be detected can be identified according to whether the marked components are detected, the content and the ratio of the component contents, the application range is wide, and the method can be used for identifying the Panax traditional Chinese medicines or the parts thereof and can also be used for identifying the authenticity and the source of the Panax components in the Chinese patent medicine prescription.

Description

Method for identifying traditional Chinese medicine derived from Panax plant or different parts thereof and application thereof

Technical Field

The invention relates to the technical field of traditional Chinese medicine identification, in particular to an identification method of traditional Chinese medicines which are derived from panax plants or different parts of the panax plants and have similar chemical compositions and application thereof.

Background

In recent years, ginseng has remarkable advantages in the development of medical treatment and health industry, and is widely researched, developed and utilized. Different ginseng traditional Chinese medicines have different medicinal properties, and can be reasonably used in the practical application process to ensure safety and effectiveness. On the other hand, because the ginseng traditional Chinese medicine has wide market prospect and higher market value, besides ginseng, red ginseng, ginseng leaves, American ginseng, pseudo-ginseng, panax japonicus and rhizoma panacis majoris which are recorded in the Chinese pharmacopoeia (2020 edition), illegal vendors replace the feeding with other ginseng medicinal materials which are not recorded in the pharmacopoeia or different parts (such as stems and leaves and flowers) of recorded plant varieties, the safety and the effectiveness of the traditional Chinese medicine are seriously influenced. Therefore, for the ginseng traditional Chinese medicine and the products containing the ginseng traditional Chinese medicine, the precise identification of the medicinal materials is very important. After the traditional Chinese medicine decoction pieces are crushed, processed and extracted, the traditional Chinese medicine decoction pieces are difficult to distinguish through conventional character identification and microscopic identification. The thin-layer method identification of the recorded traditional Chinese medicine from the panax genus by the Chinese pharmacopoeia standard mostly uses ginsenoside as an index, but the ginsenoside is a common and main active component in the traditional Chinese medicine from the panax genus and is rich in roots, stems, leaves and flowers of various panax plants. Therefore, the identification by using a single or a few ginsenosides as indexes is difficult to accurately distinguish different ginseng traditional Chinese medicines. Therefore, the method for characterizing and identifying the saponin component in the panax traditional Chinese medicine is deeply researched, and has important significance for accurately identifying the panax traditional Chinese medicine and products thereof.

Disclosure of Invention

In order to solve the technical problems, the invention provides an identification method and application of traditional Chinese medicines which are derived from panax plants or different parts of panax plants and have similar chemical compositions.

In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:

a method for identifying traditional Chinese medicine derived from Panax plants or different parts thereof comprises detecting a sample to be detected by using notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, malonyl ginsenoside Re1, 24(R) -pseudoginsenoside F11, ginsenoside Rf, ginsenoside F3, ginsenoside Rg2, ginsenoside Ra2, ginsenoside Rb1, ginsenoside Ra1, ginsenoside Rc, ginsenoside Rb2, malonyl ginsenoside Rc, ginsenoside Ro, ginsenoside Rb3, malonyl ginsenoside Rb2, panax japonicus IV, ginsenoside Rd, panax japonicus IVRd, malonyl ginsenoside Rg, notoginsenoside Rg K and ginsenoside 3 as marker components, and identifying the source of the component to be detected according to the detection result.

The content of the above marked components is different in different panax plants and different parts of the same panax plant, the specific variety and the part of the sample to be detected can be identified according to the detected content and the ratio of the content of the above marked components, the application range is wide, and the method can be used for identifying the traditional Chinese medicine of the panax or the part thereof and can also be used for identifying the authenticity and the source of the components of the panax in the prescription of the Chinese patent medicine.

Preferably, detecting the marker component in the sample to be detected by using a high performance liquid chromatography/mass spectrometry combined analysis method;

the chromatographic conditions of the high performance liquid chromatography are as follows:

a chromatographic column: a polarity-modified octadecylsilane chemically bonded silica chromatographic column;

mobile phase a was acetonitrile and mobile phase B was 0.1% v/v formic acid in water, and a linear gradient elution was performed, which was performed as follows:

flow rate: 0.28-0.32 mL/min;

column temperature: 30-40 ℃.

The mobile phase and the linear elution program adopted by the invention can improve the separation degree of different ginsenosides and reduce the tailing phenomenon, and the peak area of partial components under the mobile phase is larger, thus being beneficial to effectively detecting each marker component.

Preferably, the column is BEH Shield RP 18. Under the chromatographic conditions of the invention, better resolution, peak type and peak number of chromatographic peaks can be achieved by adopting the chromatographic column.

Preferably, the column temperature is 40 ℃. Under the chromatographic condition of the invention, more peak output numbers and better separation degree can be achieved at the column temperature of 40 ℃.

Preferably, the flow rate is 0.3 mL/min.

Preferably, the mass spectrometer is a Q-Trap 4500 triple quadrupole-linear ion Trap mass spectrometer.

Preferably, the ion source of the mass spectrum is an electrospray ion source, and data are collected in a negative ion mode; the parameters of the ion source are: spray voltage (Spray voltage) was-4500V, Capillary Temperature (TEM) was 550.0 deg.C, sheath Gas (CUR) was 35.0psi, atomization voltage (Gas 1) was 55.0psi, desolvation voltage (Gas 2) was 55.0psi, collisional Gas pressure (CAD) was High, and collisional cell extension voltage (CXP) was-13.0V.

Preferably, the identification method further comprises analyzing the mass spectrum detection result of the sample to be detected by using an MRM acquisition mode. The specific method comprises the following steps:

the ginseng has ginsenoside Rg1 (hereinafter referred to as Rg1), ginsenoside Re (hereinafter referred to as Re), malonyl ginsenoside Re1 (hereinafter referred to as m-Re1), ginsenoside Rb1 (hereinafter referred to as Rb1), ginsenoside Rc (hereinafter referred to as Rc), malonyl ginsenoside Rc (hereinafter referred to as m-Rc), ginsenoside Ro (hereinafter referred to as Ro), ginsenoside Rb3 (hereinafter referred to as Rb3), malonyl ginsenoside Rb2 (hereinafter referred to as m-Rb2), ginsenoside Rd (hereinafter referred to as Rd), malonyl ginsenoside Rd, ginsenoside Rf (hereinafter referred to as Rf), and ginsenoside Rb2 (hereinafter referred to as Rb2), does not have 24(R) -pseudoginsenoside F11 (hereinafter referred to as p-F11), and Rf/ginsenoside IV (hereinafter referred to as chiku-IV) >50 (hereinafter referred to the ratio of the two types of saponin M peaks), notoginsenoside R1 (hereinafter referred to as noto-R1)/Ro <1, and Rd/Re < 0.5;

the ginseng leaves have Rg1, Re, m-Re1, Rb1, Rc, Rb2, Rb3, Rd and m-Rd peaks, almost no p-F11 is contained, Rg1/Rb1>1, and most of the ginseng leaves are also represented by Rc/Rg1<2, Rb1/Rg1<0.5, Rc/Rb3> 5;

the ginseng flower has Rg1, Re, m-Re1, Rb1, Rc, Rb2, Rd and m-Rd peaks, the Rf content is low or almost free, and Rc/Rg1>2, Rb1/Rg1>1, Rg 1/not-R1 >1, Rb 3/not-R1 <1, m-Re 1/not-R1 >5, m-Rd/not-R1 > 10;

red ginseng has Rg1, Re, Rf, Rb1, Rc, Ro, Rd, Rf, and Rb2 peaks, and Rd/Re > 0.5;

american ginseng has Re, Rb1, m-Rb1, Rc, Ro, Rd, m-Rd and p-F11 peaks, Rg1, Rf and Rb2 contents are very low, and Rf/chiku-IV <0.5, p-F11/chiku-IV >15, noto-R1/Ro <0.5, p-F11/Rb1<1, Rb1/Rb2>1, Rb1/Rc >1, Rb1/Rb3> 1;

american ginseng leaves have noto-R1, Re, Rb1, Rc, Rb2, Rb3, m-Rb2, Rd, m-Rd and p-F11 peaks, and p-F11/Rb1>10, Rb1/Rb2<1, Rb1/Rc <1, Rb1/Rb3<1, Rb2/Rg1<12, Rc/Rb3<1, most American ginseng also shows Rc/Rg1<4, Rb3/Rg1<30, 0.6< Rg1/Rb1< 1;

american ginseng flowers have noto-R1, Re, Rb1, Rc, Rb2, Rb3, m-Rb2, Rd, m-Rd, p-F11 peaks, and p-F11/Rb1>10, Rb1/Rb2<1, Rb1/Rc <1, Rb1/Rb3<1, Rc/Rg1>5, Rb2/Rg1>14, Rb3/Rg1>30, Rg1/noto-R1<1, 10< Rb3/noto-R1<15, and most American ginseng flowers also exhibit Rb3/Rc < 50;

pseudo-ginseng has Rg1, Re, F3, Rb1, Rd, not-K and not-R1 chromatographic peaks, almost does not contain Ro, Rc and Rb2, and not-R1/Ro >1000, and most of the pseudo-ginseng also shows that 1< Rf/chiku-IV < 10;

the notoginseng leaf has Ra2, Rb1, Ra1, Rc, Rb2, m-Rc, m-Rb2, Rd and Rb3 peaks, almost no p-F11, Rc/Rb3<1, Rg1/Rb1<0.1, and most of the notoginseng leaf also shows Rb1/Re <160, Rc/Re <600, Rb3/Re < 900;

notoginseng has Ra2, Rb1, Ra1, Rc, Rb2, Rb3 and Rd peaks, and Rg 1/not-R1 <1, Rb3/Rc >100, m-Re 1/not-R1 <0.1, m-Rd/not-R1 <2, most of Notoginseng also exhibits Rb1/Re >450, Rc/Re >800, Rb3/Re >1000, Rb 3/not-R1 > 190;

the panax japonicus contains OA type saponins Ro, chiku-IV and chikun saponins IVa (hereinafter abbreviated as chiku-IVa), and Ro/chiku-IV is less than 2, most panax japonicus is also represented as chiku-IVa/Ro <1, Re/Rb1> 1;

the Panax japonicum contains OA type saponins of Ro, chiku-IV and chiku-IVa, and most Panax japonicum also has chiku-IVa/Ro >1, Ro/chiku-IV >2 and Re/Rb1< 1.

The invention also provides the application of the identification method of the traditional Chinese medicine from the panax plants or different parts thereof in identifying the sources of the panax traditional Chinese medicines in the preparation of the formula. The above detection method can identify specific Panax plant and part of the sample, and can also identify the authenticity and source of Panax component even in complex system of Chinese patent medicine.

The invention has the beneficial effects that: the method provided by the invention combines the MRM acquisition mode with good linear range with the difference analysis technology, and the specific 23 marker components can be used for realizing the comparison and the differentiation of the interspecies difference, thereby providing an effective basis for the identification of the varieties and the parts of the ginseng traditional Chinese medicines and the identification of the authenticity of the varieties of the ginseng traditional Chinese medicines in the Chinese patent medicines.

Drawings

The invention will be further described with reference to the accompanying drawings and examples, in which:

FIG. 1 is a flow chart of the prediction of OA-type molecule in example 1 of the present invention;

FIG. 2 is a PCA score plot based on Full MS data for 95 batches of Panax samples in example 1 of the present invention;

FIG. 3 is an OPLS-DA score plot based on Full MS data for 95 batches of Panax samples in example 1 of the present invention;

FIG. 4 is a graph of a displacement test based on Full MS data from 95 batches of Panax samples according to example 1 of the present invention;

FIG. 5 is a VIP graph based on Full MS data from 95 batches of Panax samples in example 1 of the present invention;

FIG. 6 is a multi-channel MRM map of ginseng in embodiment 1 of the present invention;

FIG. 7 is a multichannel MRM chart of Red Ginseng according to example 1 of the present invention;

FIG. 8 is a multi-channel MRM chart of American ginseng in example 1 of the present invention;

FIG. 9 is a multi-channel MRM map of Panax notoginseng in example 1 of the present invention;

FIG. 10 is a multichannel MRM chart of leaves of Panax ginseng in example 1 of the present invention;

FIG. 11 is a multichannel MRM chart of ginseng flowers in example 1 of the present invention;

FIG. 12 is a multichannel MRM chart of leaves of American ginseng in accordance with example 1 of the present invention;

FIG. 13 is a multichannel MRM map of American ginseng flowers in example 1 of the present invention;

FIG. 14 is a multichannel MRM map of leaves of Panax notoginseng in example 1 of the present invention;

FIG. 15 is a multichannel MRM chart of Notoginseng flower in example 1 of the present invention;

FIG. 16 is a multi-channel MRM chart of Panax japonicus in example 1 of the present invention;

FIG. 17 is a multi-channel MRM map of a reference bead in example 1 of the present invention;

FIG. 18 is a chromatogram of a characteristic marker in 8 Chinese patent drugs based on the MRM method in example 2 of the present invention;

FIG. 19 is a PCA diagram obtained by analysis of a QTrap 4500 mass spectrometer in comparative example 1 of the present invention using a pseudo-targeted metabolomics approach;

FIG. 20 is a PCA plot obtained from analysis of a Q-Orbitrap mass spectrometer of comparative example 1 of the present invention using a non-targeted metabolomics approach;

FIG. 21 is a PCA score plot based on MRM data for 95 batches of Panax samples in comparative example 1 of the present invention;

FIG. 22 is an OPLS-DA score plot based on MRM data for 95 batches of Panax samples in example 1 of the present invention;

FIG. 23 is a graph of a displacement test based on MRM data from 95 batches of Panax samples in example 1 of the present invention;

FIG. 24 is a VIP graph based on MRM data from 95 batches of Panax species samples in example 1 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Aiming at the problem that the ginseng belongs to the traditional Chinese medicines with various varieties and similar chemical compositions, and is difficult to accurately identify and distinguish, the experiment researches various ginseng traditional Chinese medicines (ginseng, red ginseng, American ginseng, pseudo-ginseng, panax japonicus, rhizoma panacis japonici, rhizoma panacis majoris, ginseng leaves, ginseng flowers, American ginseng leaves, American ginseng flowers, pseudo-ginseng leaves and pseudo-ginseng flowers), constructs a quasi-target metabonomics technology based on an ultra-high performance liquid chromatography/triple quadrupole-linear ion trap mass spectrometer, combines an MRM acquisition mode with good linear range and a differential analysis technology to form a quasi-target metabonomics strategy, directly observes the peak area under each ion pair channel, intuitively and truly reflects the content difference of intraspecific and interspecific saponins, and firstly carries out differential component analysis on five medicinal materials which are included in the ginseng traditional Chinese medicines and are not included but are similar in chemical compositions, difficult to distinguish and are easy to replace, the seven ginseng source traditional Chinese medicines recorded in the pharmacopoeia and the chemical markers which are different among the five medicinal materials which are not recorded in the pharmacopoeia but are easily replaced and used are found, and the comparison and the differentiation of the interspecies difference can be realized.

Reagents and drugs used in the following examples:

acetonitrile (Fisher, Fair lawn, NJ, USA), methanol (Fisher, Fair lawn, NJ, USA), formic acid Fisher, Fair lawn, NJ, USA) are all LC-MS grades. Deionized water was purified by a Milli-Q system (Millipore, Bedford, MA, USA).

The 60 controls were mainly divided into:

18 protopanaxatriol-type ginsenosides (PPT): ginsenoside F1 (hereinafter referred to as F1), 20(S) -ginsenoside Rh1 (hereinafter referred to as Rh1), 20(R) -ginsenoside Rh1 (hereinafter referred to as 20(R) -Rh1), ginsenoside F3 (hereinafter referred to as F3), 20(S) -notoginsenoside A3 (hereinafter referred to as A3), ginsenoside F5 (hereinafter referred to as F5), notoginsenoside R2 (hereinafter referred to as R2), 20(R) -notoginsenoside R2 (hereinafter referred to as 20(R) -R2), ginsenoside Rg2 (hereinafter referred to as Rg2), ginsenoside Rf (hereinafter referred to as Rf), ginsenoside Rg1 (hereinafter referred to as Rg1), notoginsenoside R1 (hereinafter referred to as noto-R1), notoginsenoside Fp1 (hereinafter referred to as Fp1), ginsenoside Re (hereinafter referred to as Re), 20-O-glucosyl-ginsenoside-R1-Glu), ginsenoside Re2 (hereinafter referred to as Re2), ginsenoside Re3 (hereinafter referred to as Re3), malonyl ginsenoside Re1 (hereinafter referred to as m-Re1), and ginsenoside Rb1 (hereinafter referred to as Rb 1);

22 protopanaxadiol-type ginsenosides (PPD): ginsenoside Rh2 (hereinafter referred to as Rh2), 20(R) -ginsenoside Rh2 (hereinafter referred to as 20(R) -Rh2), ginsenoside K (hereinafter referred to as K), ginsenoside F2 (hereinafter referred to as F2), ginsenoside Rg3 (hereinafter referred to as Rg3), 20(R) -ginsenoside Rg3 (hereinafter referred to as 20(R) -Rg3), notoginsenoside Fe (hereinafter referred to as Fe), ginsenoside Rd2 (hereinafter referred to as Rd2), notoginsenoside K (hereinafter referred to as noto-K), ginsenoside Rd (hereinafter referred to as Rd), gypenoside XVII (hereinafter referred to as XVII), malonyl ginsenoside Rd (hereinafter referred to as m-Rd), ginsenoside Rb2 (hereinafter referred to as Rb2), ginsenoside Rb3 (hereinafter referred to as Rb3), ginsenoside Rc (hereinafter referred to as malonyl ginsenoside Rc), malonyl ginsenoside Rc (Rc) and m-Rc, malonyl ginsenoside Rb2 (hereinafter referred to as m-Rb2), malonyl ginsenoside Rb1 (hereinafter referred to as m-Rb1), ginsenoside Ra1 (hereinafter referred to as Ra1), ginsenoside Ra2 (hereinafter referred to as Ra2), notoginsenoside R4 (hereinafter referred to as R4), and notoginsenoside T (hereinafter referred to as T);

4 oleanolic acid type ginsenosides (OA): chikusetsusaponin IVa (hereinafter abbreviated as chiku-IVa), chikusetsusaponin IV (hereinafter abbreviated as chiku-IV), pseudoginsenoside Rt1 (hereinafter abbreviated as Rt1), and ginsenoside Ro (hereinafter abbreviated as Ro);

two oxtriptolone-type ginsenosides: 24(R) -pseudoginsenoside Rt5 (hereinafter referred to as 24(R) -Rt5), 24(R) -pseudoginsenoside F11 (hereinafter referred to as p-F11);

other types of saponins: ginsenoside Rk3 (hereinafter abbreviated as Rk3), ginsenoside Rh4 (hereinafter abbreviated as Rh4), notoginsenoside T5 (hereinafter abbreviated as T5), ginsenoside Rg5 (hereinafter abbreviated as Rg5), ginsenoside Rg6 (hereinafter abbreviated as Rg6), ginsenoside F4 (hereinafter abbreviated as F4), ginsenoside Rk1 (hereinafter abbreviated as Rk1), 5, 6-dehydroginsenoside Rd, vietnamese ginsenoside R8 (hereinafter abbreviated as R8), 5, 6-dehydroginsenoside Rb 1;

four ginsengenin: 20(S) -protopanaxatriol, 20(S) -protopanaxadiol, oleanolic acid, 20(S),24(R) -ocroterenol aglycone.

The above controls were all from Shanghai Shidan De Biotechnology, Inc. and Chengderster Biotechnology, Inc.

The information of 95 batches of samples of ginseng root, ginseng leaf, ginseng flower, American ginseng root, American ginseng leaf, American ginseng flower, pseudo-ginseng root, pseudo-ginseng leaf, pseudo-ginseng flower, red ginseng, panax japonicus and rhizoma panacis majoris is shown in table 1.

Table 195 batch different variety medicinal material information table

The analytical instruments used in the following examples:

QTrap 4500 triple quadrupole-linear ion trap mass spectrometer (Applied Biosystems-SCIEX Scientific, Concord, Canada).

Example 1

The embodiment of the invention provides an obtaining process of a difference identification marker.

1. Data acquisition

1.1 preparation of sample solutions

Respectively taking 25mg of each of 7 batches of samples (ginseng, red ginseng, American ginseng, pseudo-ginseng, panax japonicus and ginseng leaves) of the traditional Chinese medicines of the genus Panax, which are collected and carried by pharmacopoeia, adding 5mL of 70% methanol-water solution (V/V) for dissolving, carrying out vortex oscillation for 2min, carrying out ultrasonic extraction for 1h, cooling an extracting solution, then supplementing the weight loss with the 70% methanol-water solution (V/V), centrifuging at 14000rpm (11481g) for 10min, taking supernatant, filtering with a 0.22 mu m microporous filter membrane to obtain a sample solution of the traditional Chinese medicines of the genus Panax with the concentration of 5mg/mL, and taking 100 mu L of filtrate for LC-MS analysis.

1.2 liquid phase and Mass Spectrometry conditions

The high performance liquid chromatography analysis is completed in an I-Class ultra performance liquid system, and the chromatographic conditions are as follows:

a chromatographic column: BEH Shield RP18 (2.1X 100mm, 1.7 μm);

mobile phase: acetonitrile (A), 0.1% v/v aqueous formic acid (B);

column temperature: 40 ℃;

flow rate: 0.3 mL/min;

sample introduction amount: 3 mu L of the solution;

a linear gradient elution was performed, the procedure of which was as follows:

the mass spectrum conditions are as follows:

the ion source was an electrospray ion source, the Spray voltage (Spray voltage) was-4500V, the Capillary Temperature (TEM) was 550.0 deg.C, the sheath Gas (CURAIn Gas, CUR) was 35.0psi, the atomization voltage (Gas 1) was 55.0psi, the desolvation voltage (Gas 2) was 55.0psi, the collisional Gas pressure (CAD) was High, and the collisional cell exit voltage (CXP) was-13.0V.

By adopting the parameters, the sample injection amount is set to be 3 mu L, the seven ginseng traditional Chinese medicine samples are subjected to target collection of two collection methods of three mother ion lists, and the data analysis adopts Analyst (1.6.3) software.

1.3 the information-dependent acquisition-enhanced product ion scanning (IDA-EPI) technology is adopted to characterize and identify the ginsenoside, the MRM-IDA-EPI mode is used for detecting neutral saponin and malonylated saponin, and the MIM-IDA-EPI mode is used for detecting oleanolic acid type ginsenoside. CE values in the MIM-IDA-EPI mode are 70eV to 100eV, and CE values in the MRM-IDA-EPI mode are 70eV to 120 eV; the information-dependent acquisition response intensity is 1-2.

2. Construction of ion pairs

2.1MRM ion Pair construction

And (3) carrying out condition selection on the parent ions transmitted by the Q1 in an IDA mode, sending the parent ions into the Q3 to obtain daughter ions, confirming the components by matching the parent ions with the daughter ions, and sending the components into an ion trap for cracking. The neutral saponin and malonylated saponin ion pair is confirmed, the unified standard is that the data of seven ginseng source traditional Chinese medicine samples collected under NL (46/44) -IDA-EPI mode are considered, and the mass number in the list is found to be [ M-H [ -H ]]^-Peaks, as parent ions for the MRM mode, and each component corresponding to the secondary fragment that responds most strongly as daughter ions, construct ion pairs for MRM-IDA-EPI mode acquisition. Finally, 106 ion pairs are obtained from the neutral saponin, 23 ion pairs are obtained from malonic acylation, and the mass number of the component capable of capturing the secondary fragment is totally 150 in the MIM-IDA-EPI mode.

2.2MIM ion Pair construction

Firstly, predicting possible structures (aglycon + glycosyl + substituent) according to the known structural characteristics of the OA type saponin, and constructing an OA type saponin database. According to the known reported OA type aglycone structure, the characteristics of the saponin are totally counted as follows; the OA type saponin has been reported to be 44 in total, as shown in Table 2, the number of sugar substrates is 5, and the substituents are 4 (-CH)₂/-C₂H₄/-C₃H₆/-C₄H₈)。

Table 2 has reported information table of oleanolic acid type saponins

According to the above rules, the invention summarizes the number of the glycosyl groups and the number of the substituent groups contained in the structures of 44 saponins, and predicts the glycosyl groups and the substituent groups on the basis of the known original structure of oleanolic acid type saponin: setting a molecular design rule that the number of 5 glycosyl groups connected to each oleanolic acid type aglycone in the prediction process is at least 1 and at most 6, the number of non-glycosyl groups is at most 2 and at least 1, and finally, the predicted number of glycosyl groups and the number of substituent groups both contain the existing number in the known saponin structure and meet the condition of the rule.

According to the rule, the OA type saponin database is expanded and predicted, 8905 molecular formulas are constructed in total, wherein the molecular formulas comprise known oleanolic acid type saponin, all predicted mass numbers are subjected to de-duplication, and 338 different mass numbers are finally obtained and used as a list of OA type ginsenoside prediction parent ion input MIM methods. A flow chart for molecular prediction of type OA is shown in FIG. 1.

The mass numbers collected by the MRM and the MIM are used as parent ions, fragment ions with the highest secondary response are used as daughter ions, and the obtained results are combined and deduplicated to finally obtain 252 ion pairs. Inputting the ion pairs into an MRM method, and constructing a quasi-target metabonomics technology for analyzing the ginseng multi-source multi-variety multi-site medicinal material difference analysis method.

3. Sample analysis

3.1 preparation of test solutions

Respectively taking 25mg of each of 12 different-variety different-batch sample powders in the table 1, adding 5mL of 70% methanol-water solution (V/V) for dissolving, performing vortex oscillation for 2min, performing ultrasonic extraction for 1h, cooling an extracting solution, then complementing weight loss by using 70% methanol-water solution (V/V), centrifuging at 14000rpm (11481g) for 10min, taking a supernatant, filtering through a 0.22 mu m microporous filter membrane to obtain a sample solution with the concentration of 5mg/mL, and taking 100 mu L of filtrate for LC-MS analysis. The conditions of liquid phase and mass spectrum are the same as 1.1, and only a detection scanning mode is needed without triggering an EPI mode.

Construction of 3.212 Chinese medicinal materials of Panax

By a pseudo-targeted metabonomics method, the method of applying the MRM method to identifying the differential components of the ginseng-derived traditional Chinese medicines is established in the embodiment, the developed process is used for analyzing the differential component analysis of 12 ginseng-derived traditional Chinese medicines, and then the respective identification points of the 12 ginseng-derived traditional Chinese medicines are found, so that the differential identification marker of each medicinal material is constructed.

3.2.1 Pattern recognition-based discovery and Structure identification of differential Saponin Compounds

Analyzing the PCA score chart (shown in figure 2), the OPLS-DA score chart (shown in figure 3), the displacement test chart (shown in figure 4) and the VIP chart (shown in figure 5) of 12 medicinal materials of panax ginseng, panax ginseng leaves, panax ginseng flowers, panax notoginseng roots, panax notoginseng leaves, panax notoginseng flowers, red ginseng, panax japonicus, rhizoma panacis japonici, rhizoma panacis majoris, and the like by using SIMCA-P14.1 software and adopting a multivariate statistical analysis method, analyzing data by using the OPLS-DA and searching potential ginsenoside markers by using the VIP chart. The data of the sample to be tested in the PCA score map are clustered well, which shows that the data are stable and reliable. In fig. 3, the result is that ginseng, red ginseng and American ginseng are gathered into a cluster, pseudo-ginseng root, pseudo-ginseng leaf and pseudo-ginseng flower are gathered into a cluster, ginseng leaf, American ginseng leaf, ginseng flower and American ginseng flower are gathered into a cluster, and panax japonicus are still gathered into a cluster. From the results, the difference between different source varieties is obvious, the distance between homologous different parts is short, and the difference of chemical components is small. Therefore, the method has good effect when being applied to metabonomics analysis of various medicinal materials with multiple sources. This example sets up that VIP >1 identifies a total of 38 differential components. This example identifies these differential components based on standard controls (retention time and mass spectral information), secondary fragmentation analysis, and the order of appearance of the binding compounds reported in the literature, with the results shown in table 3.

TABLE 3 information Table of 38 potential marker components determined based on negative ion mode MRM data

Note:^aindicating that the test sample is identified by a control sample. Not labeled^aThis indicates that the sample was not identified by the control and is the result of the estimated identification.

3.2.2 construction of identification marker for ginsenoside in Chinese medicine of Panax

According to the identified difference components, peak marking is carried out in an original MRM spectrogram, and the difference among 12 medicinal materials is more intuitively shown, as shown in figures 6-17. All the medicinal materials are compared in groups, and the internal standard substance in each group is selected from the marked difference components, so that the content difference among different varieties can be conveniently and visually represented. Analyzing through the peak area ratio of the difference components to the internal standard, summarizing the difference identification points of the traditional Chinese medicine from the genus Panax, and finding the difference identification markers of each medicinal material.

(1) Human participated in red ginseng:

common peaks of ginseng: rg1, Re, m-Re1, Rb1, Rc, Rb2, m-Rc, Ro, Rb3, m-Rb2, Rd, m-Rd and the like, and the characteristic peaks are Rf and Rb2, as shown in FIG. 6. Common peaks of red ginseng: rg1, Re, Rf, Rb1, Rc, Ro and Rd, and the characteristic peaks are Rf and Rb2, as shown in FIG. 7.

The abundance of Rg1, Re, Rf, Rb2 and Ro contained in the ginseng is high, and the two are mainly distinguished in that the malonic acid acylated ginsenoside and some small polar saponins (such as Rg3 and Rg 2); in addition, the red ginseng has higher Rd content, wherein Rd/Re in the ginseng is less than 0.5(7 batches meet, 0.29-0.47), and Rd/Re in the red ginseng is more than 0.5(10 batches meet, 0.53-1.34).

(2) Root system: ginseng, American ginseng and pseudo-ginseng

Common peaks of ginseng: rg1, Re, m-Re1, Rb1, Rc, Rb2, m-Rc, Ro, Rb3, m-Rb2, Rd, m-Rd and the like, and the characteristic peaks are Rf and Rb 2. Common peaks of american ginseng: re, Rb1, m-Rb1, Rc, Ro, Rd, m-Rd and the like, and the characteristic peak is p-F11, as shown in FIG. 8. Common peaks of notoginseng: rg1, Re, F3, Rb1, Rd, noto-K, etc., and the characteristic peak is noto-R1, and Ro, Rc, Rb2 are hardly contained, as shown in FIG. 9.

The ginseng contains rich Rg1, Re, Rf, Rb2 and Ro and does not contain p-F11, wherein Rf/chiku-IV in the ginseng is more than 50(10 batches are all satisfied, the ratio range is 70-1000), Rf/chiku-IV in the American ginseng is less than 0.5(10 batches are all satisfied, the ratio range is 0.01-0.3), and 1< Rf/chiku-IV in the panax notoginseng is less than 10 (8 batches are satisfied in 10 batches, the ratio range is 5-9.2); the content of Rg1, Rf and Rb2 in the American ginseng is very low, and the characteristic components are p-F11, p-F11/chiku-IV >15(10 batches are all satisfied, 17.1-106.3); pseudo-ginseng contains abundant noto-R1, wherein the noto-R1/Ro >1000(10 batches are all satisfied, the ratio range is 1490-2900), the noto-R1/Ro <1 in the ginseng (9 batches are satisfied, 0.2-0.9 in 10 batches), the noto-R1/Ro <0.5 in the American ginseng (10 batches are all satisfied, 0.06-0.23) and the Ro, Rc and Rb2 in the pseudo-ginseng are very low.

(3) Ginseng: root/leaf/flower

Ginseng shares peaks: rg1, Re, m-Re1, Rb1, Rc, Rb2, m-Rc, Ro, Rb3, m-Rb2, Rd, m-Rd, etc.; the characteristic peak is Rf; common peaks of ginseng leaves: rg1, Re, m-Re1, Rb1, Rc, Rb2, Rb3, Rd, m-Rd and the like, and the Rf content is low, as shown in FIG. 10; ginseng flower common peaks: rg1, Re, m-Re1, Rb1, Rc, Rb2, Rd, m-Rd, etc., with low Rf content, as shown in FIG. 11.

The ginseng root is rich in Rf, Rb1, Ro, Rc and Rb2, and has medium content of m-Rb1, m-Rc and m-Rb 2; leaves and flowers are rich in Rg1, Re, Rc, Rb1 and malonylated ginsenosides m-Re1 and m-Rd, but are low in content or almost free of Ro, Rf and Rb 3. The content of m-Re1, Rc and Rb1 in the ginseng flower is higher in combination with peak area, wherein Rc/Rg1>2(5 batches meet all, 2.8-3.3) and Rb1/Rg1>1(5 batches meet all, 1.7-2.3); Rc/Rg1<2 (9 batches of 10 are satisfied, 0.7-1.7), Rb1/Rg1<0.5 (9 batches of 10 are satisfied, 0.34-0.47) in ginseng leaf.

(4) American ginseng: root/leaf/flower

Common peaks of american ginseng: re, Rb1, m-Rb1, Rc, Ro, Rd, m-Rd and the like, and the characteristic peak is p-F11; common peaks of American ginseng leaves: the content of the p-F11 is rich, such as noto-R1, Re, Rb1, Rc, Rb2, Rb3, m-Rb2, Rd, m-Rd and the like, as shown in figure 12; common peaks of American ginseng flowers: noto-R1, Re, Rb1, Rc, Rb2, Rb3, m-Rb2, Rd, m-Rd, p-F11, as shown in FIG. 13.

The characteristic components of the American ginseng are p-F11, Rb1 is rich, and contains moderate Rd and m-Rd, but the content is very low or does not contain Rg1, Rf and Rb 2; american ginseng leaves and flowers, which are richer in p-F11 compared with roots, p-F11/Rb1<1 in roots (10 lots are all satisfied, 0.07-0.23), and p-F11/Rb1>10 in leaves and flowers (10 lots are all satisfied, 12.6-44.7); the American ginseng root contains abundant Rb1, and the contents of three PPD type saponins of Rc, Rb2 and Rb3 in leaves and flowers are higher, the chromatographic peak heights of the American ginseng root are Rb1/Rb2, Rb1/Rc and Rb1/Rb3 are all greater than 1, and the chromatographic peak heights of the stems, leaves and flowers are all Rb1/Rb2, Rb1/Rc and Rb1/Rb3 are all less than 1; compared with the flowers, the American ginseng leaves have richer contents of Rc, Rb2 and Rb3 in the flowers, Rc/Rg1>5(5 batches all meet, 5.3-15.7), Rb2/Rg1>14(5 batches all meet, 14.8-22.1), Rb3/Rg1>30(5 batches all meet, 32.6-53.4), Rc/Rg1<4 (4 batches in 5 meet, the ratio range is 0.7-3.4), Rb2/Rg1<12(5 batches all meet, 2.6-11.8) and Rb3/Rg1<30 (4 batches in 5 meet, 10.9-24.2) in the American ginseng flowers.

(5) Pseudo-ginseng: root/leaf/flower

Common peaks of notoginseng roots: rg1, Re, F3, Rb1, Rd, noto-K, etc., characterized by high levels of noto-R1, but low or essentially no levels of Ro and Rc; common peaks of panax notoginseng leaves: ra2, Rb1, Ra1, Rc, Rb2, m-Rc, m-Rb2, Rd and the like, wherein the content of Rb3 is high, but the contents of noto-R1, Rg1 and Re are low, as shown in FIG. 14; common peaks of notoginseng flowers: ra2, Rb1, Ra1, Rc, Rb2, Rb3 and Rd, wherein the Rc content is rich, but the noto-R1, Rg1 and Re content are low, as shown in FIG. 15.

The pseudo-ginseng root is rich in noto-R1 and Rg1, but has low Ro, chiku-IV and Rb2 contents; the pseudo-ginseng leaf and pseudo-ginseng flower are rich in Rc and Rb3, have moderate Rd content, and also contain Ra1 and Ra2, but have very low noto-R1 content. Compared with the pseudo-ginseng leaf, the content of Rb1, Rc and Rb3 in the pseudo-ginseng flower is higher, Rb1/Re >450 (4 in 5 batches meet 467.3-1001.8), Rc/Re >800 (4 in 5 batches meet 825.7-1611.9), Rb3/Re >1000 (4 in 5 batches meet 1205.3-2559.4); rb1/Re <160 in leaves (4 in 5 lots, 54.7-157.7), Rc/Re <600 (4 in 5 lots, 187.6-595.8), and Rb3/Re <900 (4 in 5 lots, 350.1-885.6).

(6) Rhizoma Panacis Japonici and rhizoma Panacis Majoris

Compared with other ginseng traditional Chinese medicines, the panax japonicus is rich in OA-type saponins Ro, chiku-IV and chiku-IVa and is a characteristic component. The content of chiku-IVa in the panax japonicus is richer, namely chiku-IVa/Ro is greater than 1 (7 of 10 batches meet, 1.03-3.2), while chiku-IVa/Ro in the panax japonicus is less than 1(8 of 10 batches meet, 0.4-0.7); the chiku-IV and Re contents in the panax japonicus are higher, Ro/chiku-IV <2(10 batches all meet, 0.1-1.7), and Ro/chiku-IV >2 in the panax japonicus (7 batches meet, 2.0-4.2 in 10 batches); Re/Rb1>1 in Panax japonicus (9 out of 10 batches are satisfied, 2.3-24.1), and Re/Rb1<1 in Panax japonicus (7 out of 10 batches are satisfied, 0.05-0.9). As shown in fig. 16 and 17.

(7) Leaves: folium Ginseng/folium Panacis Quinquefolii/folium Notoginseng

Common peaks of ginseng leaves: rg1, Re, m-Re1, Rb1, Rc, Rb2, Rb3, Rd, m-Rd and the like, wherein Rg1 and m-Re1 are high in content, but p-F11 is low in content; common peaks of American ginseng leaves: noto-R1, Re, Rb1, Rc, Rb2, Rb3, m-Rb2, Rd, m-Rd, etc.; is rich in p-F11; common peaks of panax notoginseng leaves: ra2, Rb1, Ra1, Rc, Rb2, m-Rc, m-Rb2, Rd, etc.; but the content of Rg1 and p-F11 is low.

The ginseng leaf contains Rg1 and m-Re1, and the American ginseng leaf contains rich p-F11 and m-Rd; rb3 and Rc are Rc/Rb3>5 in ginseng leaf (7 of 10 batches are satisfied, 5.75-8.16), and Rc/Rb3<1 in American ginseng stem and leaf and notoginseng leaf (10 batches are satisfied, 0.08-0.68); the ginseng leaf and the pseudo-ginseng leaf hardly contain p-F11, and the p-F11/Rb1 in the American ginseng leaf is more than 10(5 batches completely meet, 15.0-44.7); the content of Rg1 in the leaves of American ginseng and the leaves of pseudo-ginseng is very low, 0.6< Rg1/Rb1<1 (4 batches of 5 are satisfied, 0.65-0.93), Rg1/Rb1<0.1(5 batches are satisfied, 0.01-0.02), and the leaves of ginseng 1/Rb1>1(10 batches are satisfied, 1.7-3.2); in addition, the notoginseng leaf contains Ra1 and Ra2, which are contained in the ginseng leaf and the American ginseng leaf in a very low content.

(8) Flowers: ginseng flower/American ginseng flower/Notoginseng flower

Ginseng flower common peaks: rg1, Re, m-Re1, Rb1, Rc, Rb2, Rd, m-Rd and the like, wherein the content of m-Re1 is high, but the content of p-F11 is low; common peaks of American ginseng flowers: p-F11 and m-Rb2 with high contents such as noto-R1, Re, Rb1, Rc, Rb2, Rb3, m-Rb2, Rd, m-Rd and the like are contained; common peaks of notoginseng flowers: ra2, Rb1, Ra1, Rc, Rb2, Rb3, Rd and the like, but the content of p-F11 is low.

The ginseng flower is rich in Rg1, Re and m-Re1, Rg1/noto-R1>1(5 batches are all satisfied, 1.23-2.0); but Rg1/noto-R1<1(10 batches are all satisfied, 0.2-0.5) in the American ginseng flower and the sanchi flower; rb3 is rich in Notoginseng flower, but has low content in flos Ginseng, and flos Ginseng Rb3/noto-R1<1 (all of 5 batches meet, 0.6-0.9); 10< Rb3/noto-R1<15 in American ginseng flowers (5 batches all meet, 10.7-14.9); and Rb3/noto-R1>190 in sanchi flower (5 batches all meet, 192.8-353.0); the American ginseng flower contains high content of p-F11 as characteristic components, and contains Rb3 and m-Rb 2; and American ginseng flower Rb3/Rc <50 (4 of 5 batches meet, ratio range 28.3-46.7), and Notoginseng flower Rb3/Rc >100(5 batches all meet, ratio range 103.3-364.3); notoginseng flower contains almost no Rg1 and Re, has relatively low content of malonylated saponin, m-Re1/noto-R1<0.1(5 batches meet all, 0.01-0.02) and m-Rd/noto-R1<2(5 batches meet all, 1.1-1.9); and m-Re 1/not-R1 >5(5 batches are all satisfied, 5.01-8.0) and m-Rd/not-R1 >10(5 batches are all satisfied, 10.9-17.0) in the ginseng flower respectively; the ginseng flower is rich in Ra1, Ra2 and Rb 3.

In the comparison of all the different components, 23 components are mentioned, and are respectively: noto-R1(977.4-931.3), Rg1(845.4-475.3), Re (991.4-799.4), m-Re1(1031.6-945.5), p-F11(845.7-799.3), Rf (845.7-799.3), F3(815.3-475.3), Rg2(829.5-637.3), Ra2(1255.4-1209.6), Rb1(1153.6-1107.5), Ra1(1255.4-1209.6), Rc (1123.3-1077.6), rb2(1123.3-1077.6), m-Rc (1163.3-1077.7), Ro (955.5-793.4), Rb3(1123.3-1077.6), m-Rb2(1163.3-1077.7), chiku-IV (925.5-613.3), Rd (991.1-945.5), chiku-IVa (793.4-569.4), m-Rd (1031.6-945.5), noto-K (991.1-945.5), Rg3(829.5-459.4), and the 23 components can be used as difference identification markers of 12 medicinal materials.

Example 2

In this example, 29 Chinese patent medicines of different dosage forms containing ginseng-derived Chinese medicines were selected, as shown in table 4.

TABLE 429 batch Chinese patent medicine information table

Selecting the following 14 ion channels to detect the ginsenoside markers: noto-R1(977.4-931.3), Rg1(845.4-475.3), Re (991.4-799.4), p-F11(845.7-799.3), Rf (845.7-799.3), F3(815.3-475.3), Rg2(829.5-637.3), Rb1(1153.6-1107.5), Rc (1123.3-1077.6), Rb2(1123.3-1077.6), Ro (955.5-793.4), Rb3(1123.3-1077.6), Rd (991.1-945.5) and Rg3(829.5-459.4), wherein the types of traditional Chinese medicines from Panax in the Chinese patent medicine are identified by the ion pair marker components. By setting the ion pair information in the MRM mode, different components of the Chinese patent medicines are captured in different types of Chinese patent medicines so as to verify the capability of the difference identification marker in the authenticity identification of the Chinese patent medicines. As shown in fig. 18 (the corresponding Chinese patent medicines for the abbreviations are shown in table 4), for example, the Chinese patent medicine containing ginseng and red ginseng has more abundant components, while the Chinese patent medicine containing American ginseng and pseudo-ginseng has relatively less components. The contents of Rf, Rc, Rb2 and Rb3 are relatively high, which possibly indicates that ginseng Yangrong pills, Shenling Baizhu powder, Guiling and Shengmai drink may contain ginseng, and the Guiling and Shengmai drink contains relatively rich small-polarity saponins Rg2 and Rg3, but the ginseng does not contain ginseng, thus proving that the two Chinese patent medicines contain red ginseng; rare Rg1, Rf and Rb2 and rich p-F11 can prove that American ginseng lung-protecting pills and twenty-seven-ingredient women pills contain American ginseng; the noto-R1 and Rg1 are rich and have rare Rf, Rc and Rb2, and the powder for promoting blood circulation and relieving pain and the capsule for treating the blood injury of pseudo-ginseng are proved to contain pseudo-ginseng. The method can be used for identifying Panax Chinese medicinal species in other 21 Chinese patent medicines, as shown in Table 5.

TABLE 529 Chinese patent medicine characteristic component information table

It should be noted that the minipill can be found to contain rich noto-R1, Rg1, Re, Rg2 and Rg3, and red ginseng and pseudo-ginseng which are characteristic components of red ginseng and pseudo-ginseng at the same time, so that the minipill is proved to contain the red ginseng and the pseudo-ginseng at the same time.

Through the results, the differential component positioning result obtained by the traditional Chinese medicine from the panax origin is verified in the traditional Chinese medicine sample, and the MRM method is proved to be capable of visually describing the types and distribution conditions of ginsenoside in the traditional Chinese medicine and the traditional Chinese medicine, and simultaneously distinguishing the traditional Chinese medicine from the panax origin, so that the application range is wide, and the method can be applied to true and false identification of the traditional Chinese medicine prescription.

Comparative example 1

The comparative example is a comparative study in the difference analysis of Chinese medicinal herbs in ginseng based on QTrap-MS quasi-targeted metabonomics and Q-Orbitrap-MS non-targeted metabonomics technology.

The MRM acquisition mode has no data information of Full MS, so that the MRM acquisition mode cannot be analyzed by QI software. Therefore, in this embodiment, a manual recording mode is adopted, chromatographic peaks under different retention times in each channel are labeled, retention time and ion pair information are input into Multi Quat software, 483 chromatographic peaks are labeled in total, uniform integration processing is performed on all batches of samples, and a list of ion pair-peak area data is derived. The data results are processed by 80% rule and 30% variation and then introduced into SIMCA-P14.1 software for multivariate statistical analysis. Two groups of data are shown in fig. 19 and 20, fig. 19 is a PCA diagram obtained by analyzing a QTrap 4500 mass spectrometer by a quasi-targeted metabonomics method, and fig. 20 is a PCA diagram obtained by analyzing a Q-Orbitrap mass spectrometer by a non-targeted metabonomics method. As can be seen from the figure, the two methods of the two instruments are different from each other in clustering situations of different kinds of medicinal materials. The QC data of the ginseng and the red ginseng are clustered well, the stability of the collected data is shown to be good, the left image is divided into five communities, wherein the ginseng, the red ginseng, the panax japonicus and the panax japonicus are clustered together respectively, and the American ginseng, the pseudo-ginseng and the ginseng leaf are divided into three independent communities, so that the difference of chemical compositions between the ginseng and the red ginseng processed by the ginseng and the panax japonicus is relatively small. Although the ginseng leaf and the ginseng are the same variety, the ginseng leaf and the ginseng have a certain distance and have a significant difference due to different parts, and the distance between the panax notoginseng and the ginseng is relatively far in a PCA diagram, which proves that the significant difference also exists. According to the data of the right figure, three communities are totally divided, wherein ginseng, ginseng leaves, American ginseng and red ginseng are gathered together, panax japonicus and rhizoma panacis majoris are gathered together, and pseudo-ginseng is independently gathered into a cluster. Although the difference between different varieties is large, the difference between similar varieties such as ginseng and ginseng leaves is small. Therefore, the analysis is carried out according to the clustering degree of the data, and the application of the quasi-targeted metabonomics technology has greater advantages for the difference analysis between different varieties in the same genus.

Next, in this example, two sets of metabonomics methods were analyzed, and OPLS-DA further analysis was performed, and the VIP graph found the main difference components thereof, and the displacement test graph demonstrated that the two sets of models did not over-fit, as shown in fig. 2 to 5 and fig. 21 to 24. Wherein the VIP value of >1.5 under the non-targeted metabonomics method has 41 components in total, 29 components are identified (or derived) according to the standard substance and the secondary fragment in total, the VIP value of >1 under the pseudo-targeted metabonomics method has 50 components in total, and 38 components are identified (or derived) according to the standard substance and the secondary fragment in total. The results of the difference components obtained by the two sets of metabonomics techniques were analyzed and the linear range of peak area of 10 common components of the two sets of data was selected and counted as shown in table 6 below.

TABLE 6 comparison of Linear Range of pseudo-targeted and non-targeted metabolomics

According to the comparison result of the two groups of data, the linear range which can be captured by the MRM method is wider, the coverage is wider, the maximum value and the minimum value cover 4 orders of magnitude at most, and the Full MS method mainly covers 2 orders of magnitude and 3 orders of magnitude. Therefore, the MRM method is shown to cover a wider linear range of the difference components, and can reflect the content difference of the medicinal material components of different batches more truly.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims (8)

1. A method for identifying traditional Chinese medicines derived from Panax plants or different parts thereof is characterized in that notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, malonyl ginsenoside Re1, 24(R) -pseudoginsenoside F11, ginsenoside Rf, ginsenoside F3, ginsenoside Rg2, ginsenoside Ra2, ginsenoside Rb1, ginsenoside Ra1, ginsenoside Rc, ginsenoside Rb2, malonyl ginsenoside Rc, ginsenoside Ro, ginsenoside Rb3, malonyl ginsenoside Rb2, chikusetsusaponin IV, ginsenoside Rd, chikusetaponin IVa, malonyl ginsenoside Rd, notoginsenoside K and ginsenoside Rg3 are used as marking components to detect a sample to be detected, and the source of the component to be detected is identified through the detection result.

2. The method of claim 1, wherein the marker component is detected in the sample by HPLC/MS;

the chromatographic conditions of the high performance liquid chromatography are as follows:

a chromatographic column: a polarity-modified octadecylsilane chemically bonded silica chromatographic column;

mobile phase a was acetonitrile and mobile phase B was 0.1% v/v formic acid in water, and a linear gradient elution was performed, which was performed as follows:

flow rate: 0.28-0.32 mL/min;

column temperature: 30-40 ℃.

3. The method of claim 2, wherein the chromatographic column is BEH Shield RP 18.

4. The method of claim 2, wherein the column temperature is 40 ℃.

5. The method of claim 2, wherein the flow rate is 0.3 ml/min.

6. The method of identifying a herb derived from Panax species or different parts thereof as claimed in any of claims 1 to 5, wherein the mass spectrometry is Q-Trap 4500 triple quadrupole-linear ion Trap mass spectrometer.

7. The method of claim 6, wherein the ion source of the mass spectrometer is an electrospray ion source, and data are collected in negative ion mode; the parameters of the ion source are: the spray voltage was-4500V, the capillary temperature was 550.0 deg.C, the sheath gas was 35.0psi, the atomization voltage was 55.0psi, the desolventizing voltage was 55.0psi, the collision pressure was High, and the collision cell outlet voltage was-13.0V.

8. Use of the method of any one of claims 1 to 7 for identifying a source of a Panax species of Chinese medicinal material in a formulated product.

Images