Qualitative or quantitative analysis methods for the monosaccharide glycosidic bond linkage mode of polysaccharides in traditional Chinese medicine

Abstract

The present invention discloses a qualitative or quantitative analysis method for the glycosidic bond connection mode of monosaccharides of traditional Chinese medicine polysaccharides. The present invention uses the separation technology of GC-MS / MS to separate partially methylated sugar nitrile acetate derivatives of different glycosidic bond connection types, and obtains the qualitative and quantitative ion pairs and corresponding OCE values ​​of partially methylated sugar nitrile acetate, and constructs the qualitative and quantitative targeted MRM data array and OCE data array of partially methylated sugar nitrile acetate of multiple monosaccharides, which can perform qualitative analysis on different glycosidic bond connection modes of the same monosaccharide. Compared with the existing glycosidic bond analysis method of traditional Chinese medicine polysaccharides, it solves the problem that there are many interference peaks in the chromatogram obtained by the existing GC-MS method only through the full scan mode, resulting in many errors in the connection attribution of sugar residues in the glycosidic bond analysis, effectively eliminates the interference of by-products, improves the sensitivity and accuracy of the analysis, and has significant advantages in the qualitative and quantitative analysis between isomers of different glycosidic bond connection modes of the same monosaccharide.

Description

Technical Field

[0001] The invention relates to an analysis method for the glycosidic bond connection mode of polysaccharides, in particular to a qualitative or quantitative analysis method for the monosaccharide glycosidic bond connection mode of traditional Chinese medicine polysaccharides, and belongs to the field of glycosidic bond analysis of traditional Chinese medicine polysaccharides. Background Art

[0002] Gas chromatography-mass spectrometry (GC-MS) is the most widely used method in the field of gas chromatography. It applies the high separation ability of GC and the high identification ability of EI-MS to the identification of unknown compounds in complex systems. It plays an important role in the analysis of the connection position and order of sugar residues in traditional Chinese medicine glycoside compounds. At present, the study of glycosidic bond connection sites of traditional Chinese medicine glycoside compounds mainly relies on the full scan mode of GC-MS. This mode has poor selectivity and sensitivity, and the interference peaks in the chromatographic peaks cannot be eliminated, which increases the difficulty of glycoside connection analysis and hinders the accurate analysis of glycosidic bond connection mode of traditional Chinese medicine polysaccharides. It is urgent to improve. Summary of the invention

[0003] The main purpose of the present invention is to provide a qualitative or quantitative analysis method for the monosaccharide glycosidic bond connection mode of traditional Chinese medicine polysaccharides;

[0004] To achieve the above object, the technical solution adopted by the present invention includes:

[0005] A qualitative or quantitative analysis method for the monosaccharide glycosidic bond connection mode of traditional Chinese medicine polysaccharides:

[0006] (1) preparing partially methylated nitrile acetates of different types of monosaccharides;

[0007] (2) using GC-MS / MS method to separate different partially methylated sugar nitrile acetates and obtain EI-MS / MS spectra;

[0008] (3) determining the partially methylated sugar nitrile acetate qualitative ion pair and the partially methylated sugar nitrile acetate quantitative ion pair according to the EI-MS / MS spectrum and constructing a targeted qualitative and quantitative MRM data array in combination with the collision energy corresponding to the partially methylated sugar nitrile acetate qualitative ion pair and the partially methylated sugar nitrile acetate quantitative ion pair;

[0009] (4) Calculate the OCE values ​​of partially methylated sugar nitrile acetate ion pairs to obtain the OCE data array;

[0010] (5) Qualitative or quantitative analysis of the glycosidic bond connection patterns of Chinese medicine polysaccharides based on MRM data arrays or OCE data arrays.

[0011] As a preferred embodiment of the present invention, the different types of monosaccharides described in step (1) include but are not limited to any one or more of Glc (glucose), Man (mannose), Gal (galactose), Ara (arabinose), Xyl (D-xylose), Rib (ribose), Rha (rhamnose) or Fuc (fucose); wherein the partially methylated sugar nitrile acetate obtained from Glc has 24 Glc glycosidic bond connection modes, and the partially methylated sugar nitrile acetate obtained from Gal has 24 Gal glycosidic bond connection modes. , the partially methylated sugar nitrile acetate obtained from Man has 23 Man glycosidic bond connection modes, the partially methylated sugar nitrile acetate obtained from Ara has 12 Ara glycosidic bond connection modes, the partially methylated sugar nitrile acetate obtained from Xyl has 12 Xyl glycosidic bond connection modes, the partially methylated sugar nitrile acetate obtained from Rib has 12 Rib glycosidic bond connection modes, the partially methylated sugar nitrile acetate obtained from Rha has 8 Rha glycosidic bond connection modes, and the partially methylated sugar nitrile acetate obtained from Fuc ​​has 8 Fuc glycosidic bond connection modes.

[0012] As a preferred embodiment of the present invention, the parameters of the gas phase conditions and mass spectrometry conditions of the GC-MS / MS method in step (2) include:

[0013] The gas phase conditions included: injection temperature of 250°C; injection volume of 1 μL; split ratio of 20:1, capillary column of TG-1MS (30 m×0.25 mm×0.25 μm), and the heating program was as follows: 110°C for 2 min, gradually heated to 210°C at a rate of 5°C / min and maintained for 2 min, then heated to 280°C at a rate of 20°C / min and maintained for 2 min;

[0014] The mass spectrometry conditions included: EI source, ion source temperature of 280°C, scan rate of 0.2 scan / s, and automatic full scan mode with a scan range of m / z 50-550.

[0015] As a preferred specific embodiment of the present invention, in step (3), according to the EI-MS / MS spectrum, an ion pair with a large response intensity is selected as the quantitative ion pair of partially methylated sugar nitrile acetate, and an ion pair with structural specificity is selected as the quantitative ion pair of partially methylated sugar nitrile acetate.

[0016] As a preferred specific embodiment of the present invention, in step (3), the response intensity of the ion pair is obtained under the background of a collision energy range of 5-45 eV (variation interval is 2 eV), and the collision energy corresponding to the partially methylated sugar nitrile acetate qualitative ion pair and the partially methylated sugar nitrile acetate quantitative ion pair is determined by comparing the response intensities under different collision energies.

[0017] As a preferred specific embodiment of the present invention, the method for constructing an OCE data array by calculating the OCE value of the partially methylated sugar nitrile acetate ion pair in step (4) includes: obtaining the response intensity of the ion pair under the background of a collision energy range of 5-45eV (variation interval of 2eV), fitting the obtained data with a normal regression curve through a Guass function, and calculating the OCE value of the partially methylated sugar nitrile acetate ion pair (Q1 / Q3).

[0018] As a preferred embodiment of the present invention, in step (5), the main components of the monosaccharide of the Chinese medicine polysaccharide to be detected are first analyzed, and then the monosaccharide glycosidic bond connection mode of the Chinese medicine polysaccharide is qualitatively or quantitatively analyzed according to the MRM data array or the OCE data array.

[0019] As a preferred embodiment of the present invention, in step (5), the isomers of the same monosaccharide are qualitatively or quantitatively analyzed according to the MRM data array, or the isomers of some of the same monosaccharides and heterogeneous monosaccharides are qualitatively analyzed according to the OCE data array; more preferably, the isomeric glycoside connections in the traditional Chinese medicine polysaccharides are identified by the MRM data array and the OCE values.

[0020] As a preferred specific embodiment of the present invention, the Chinese medicinal polysaccharide includes but is not limited to any one of Auricularia auricula, Astragalus membranaceus and American ginseng polysaccharide.

[0021] The present invention adopts the GC-MS / MS separation technology to separate partially methylated sugar nitrile acetate derivatives of different glycosidic bond connection types, obtains partially methylated sugar nitrile acetate qualitative and quantitative ion pairs and corresponding OCE values, constructs qualitative and quantitative targeted MRM data arrays and OCE data arrays of partially methylated sugar nitrile acetates of multiple monosaccharides, can perform qualitative analysis on different glycosidic bond connection modes of the same monosaccharide, and solves the problem that there are many interference peaks in the chromatogram obtained only by full scanning in the existing GC-MS method, resulting in many errors in the connection attribution of sugar residues in glycosidic bond analysis, effectively eliminates the interference of by-products, improves the sensitivity and accuracy of the analysis, has significant advantages in the qualitative and quantitative analysis between isomers of the same monosaccharide with different glycosidic bond connection modes, and has broad application prospects in the glycosidic bond analysis of traditional Chinese medicine polysaccharides.

[0022] Abbreviations used in the present invention

[0023] GC-MS / MS: gas chromatography tandem mass spectrometry.

[0024] EI-MS / MS spectrum: electron impact source-mass spectrometry / mass spectrometry spectrum.

[0025] MRM data array: Multiple reaction monitoring data array.

[0026] OCE data array: optimal collision energy data array. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Comparison of the full scan chromatogram and MRM chromatogram of Gal-methylated sugar nitrile acetate.

[0028] Figure 2 Regression curve fitting results of OCE values ​​of 4,6-di-O-methyl-2,3,5-tri-O-acetyl-1-cyano-galactitol (A) and 4,6-di-O-methyl-2,3,5-tri-O-acetyl-1-cyano-glucitol (B).

[0029] Figure 3 MRM chromatogram of methylation analysis of Auricularia auricularia polysaccharide.

[0030] Figure 4 MRM chromatogram of methylation analysis of Astragalus polysaccharides.

[0031] Figure 5 MRM chromatogram of methylation analysis of American ginseng polysaccharides. DETAILED DESCRIPTION

[0032] The present invention will be further described below in conjunction with specific embodiments, and the advantages and features of the present invention will become clearer as the description proceeds. However, these embodiments are exemplary only and do not constitute any limitation to the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the scope of protection of the present invention.

[0033] Example 1 Construction of MRM data array and OCE data array for qualitative or quantitative analysis or identification of glycosidic bond connection mode of traditional Chinese medicine polysaccharides

[0034] 1. Instruments and Reagents

[0035] Thermo Trace 1300-TSQ 9000GC-MS / MS was used as the detection platform. ML104 / 02 electronic analytical balance (Mettler Toledo Instrument Shanghai Co., Ltd.); Vortex 3000 vortex shaker (Wiggens, Germany); Mikro 200R centrifuge (Scilogex, USA); KQ-500DB digital control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.). Iodomethane (Zhengzhou Alpha Chemical Co., Ltd.); barium oxide (Gade Co., Ltd.); N, N-dimethylformamide (Tianjin Fuyu Fine Chemical Co., Ltd.); barium hydroxide (Tianjin Tianli Chemical Reagent Co., Ltd.); trifluoroacetic acid (Shanghai McLean Biochemical Technology Co., Ltd.); acetic anhydride (Tianjin Alpha Biotechnology Co., Ltd.); pyridine (Tianjin Fuyu Fine Chemical Co., Ltd.); hydroxylamine hydrochloride (Tianjin Aopusheng Chemical Co., Ltd.).

[0036] 2. Test methods

[0037] This example is based on the partially methylated nitrile acetates of Glc, Gal, Man, Ara, Xyl, Rib, Rha and Fuc, and uses the Thermo Trace 1300-TSQ 9000 detection method to separate different monosaccharide derivatives, with helium as the carrier gas, a volume flow rate of 1.2 mL / min, and an interface temperature maintained at 280°C. The gas phase conditions are as follows: injection temperature is 250°C; injection volume is 1 μL; split ratio is 20:1, capillary column is TG-1MS (30m×0.25mm×0.25μm), and the heating program is as follows: 110°C for 2 min, gradually heated to 210°C at a rate of 5°C / min, and maintained for 2 min, then heated to 280°C at a rate of 20°C / min, and maintained for 2 min. Mass spectrometry conditions: EI source, ion source temperature of 280°C, scan rate of 0.2 scan / s, Auto full scan mode scanning range of m / z 50-550, detection information of Auto product ion and Auto SRM mode is shown in Table 1.

[0038] 3. Test results

[0039] Taking Gal as an example, the qualitative and quantitative targeted MRM data array of partially methylated sugar nitrile acetate is shown in Table 2. The chromatograms of Gal obtained by full scan and MRM detection modes are shown in Table 2. Figure 1 As shown, by comparison, it can be found that the MRM detection method eliminates the influence of interferences, can improve the sensitivity and accuracy of the analysis, simplifies the existing detection methods, shortens the time for data analysis, and can perform qualitative analysis on isomers of the same monosaccharide with different glycosidic bond connection methods.

[0040] Taking 4,6-di-O-methyl-2,3,5-tri-O-acetyl-1-cyano-galactitol (2,3-Galp) and 4,6-di-O-methyl-2,3,5-tri-O-acetyl-1-cyano-glucitol (2,3-Glcp) as examples, the fitting results of OCE values ​​are as follows: Figure 2 As shown, the OCE values ​​are 16.43 eV and 24.46 eV, respectively.

[0041] The results show that by comparing the OCE values, the different glycosidic bond connection modes of some isomeric monosaccharides can be qualitatively analyzed. Taking Gal as an example, the constructed OCE data array for qualitative identification of the connection analysis of some methylated sugar nitrile acetate glycosides is shown in Table 2.

[0042]

[0043]

[0044]

[0045] Experimental Example 1 Application of the MRM data array and OCE data array constructed in Example 1 in the analysis of glycosidic bond connection mode of Auricularia auricula polysaccharides

[0046] In order to accurately analyze the connection mode of glycosidic bonds in wood ear polysaccharides, this experiment first detected wood ear polysaccharides through monosaccharide composition analysis. The results showed that wood ear polysaccharides were mainly Man, with a small amount of Xyl. On this basis, the constructed partially methylated sugar nitrile acetate connection analysis qualitative and quantitative targeted MRM data array and OCE data array were applied to the analysis of glycosidic bond connection mode of wood ear polysaccharides using GC-MS / MS detection technology and TG-1MS capillary column as the platform. The analysis results are as follows: Figure 3 shown.

[0047] The characteristic ion pairs of m / z 117 / 43 and m / z 101 / 39 indicated that the molecule contained T-Xylp. The characteristic ion pairs of m / z 117 / 43 and m / z 101 / 55 indicated that the molecule contained 2-Xylp. The characteristic ion pairs of m / z 129 / 87 and m / z 161 / 43 indicated that the molecule contained T-Manp. The characteristic ion pairs of m / z 129 / 87 and m / z 101 / 68 indicated that the molecule contained 3-Manp. The characteristic ion pairs of m / z 129 / 87 and m / z 101 / 85 indicated that the molecule contained 2,3-Manp. The characteristic ion pairs of m / z 129 / 87 and m / z 169 / 109 indicated that the molecule contained 3,4-Manp. The characteristic ion pair of m / z 186 / 84 can be used to determine the presence of 3,6-Manp connection mode; the characteristic ion pairs of m / z 129 / 87 and m / z 197 / 112 can be used to determine the presence of 2,3,4-Manp connection mode; the characteristic ion pairs of m / z 129 / 87 and m / z 214 / 84 can be used to determine the presence of 2,3,6-Manp connection mode; the characteristic ion pairs of m / z 127 / 85 and m / z 184 / 66 can be used to determine the presence of 3,4,6-Manp connection mode; the characteristic ion pairs of m / z 115 / 73 and m / z 187 / 43 can be used to determine the presence of 2,3,4,6-Manp connection mode.

[0048] Among them, the OCE value of 2,3-Manp in Auricularia auricularia polysaccharide is 16.55eV, the OCE value of 2,3,6-Manp is 19.38eV, and the OCE value of 2,3,4,6-Manp is 20.61eV. By comparing the values ​​in the OCE data array of partially methylated sugar nitrile acetate glycoside connection analysis, it can be confirmed that the above analysis results are correct.

[0049] Results of qualitative and quantitative analysis of the glycosidic bond connection mode of Auricularia auricularia polysaccharide ( Figure 3 ), the results showed that the main glycosidic bond connection mode of Auricularia auriculariae polysaccharide was Man, and the percentage of Man in the polysaccharide was 94.41%, among which 3-Manp, 2,3-Manp, 3,6-Manp and 2,3,6-Manp were the main connection modes, and the percentages of Man in the polysaccharide were 37.32%, 16.77%, 15.22% and 17.37%, respectively.

[0050] Experimental Example 2 Application of the MRM data array and OCE data array constructed in Example 1 in the analysis of glycosidic bond connection mode of astragalus polysaccharides

[0051] In order to accurately analyze the connection mode of glycosidic bonds in astragalus polysaccharides, this experiment first detected astragalus polysaccharides through monosaccharide composition analysis. The results showed that astragalus polysaccharides were mainly composed of Glc and a small amount of Ara. On this basis, the constructed partially methylated sugar nitrile acetate connection analysis qualitative and quantitative targeted MRM data array and OCE data array were applied to the analysis of glycosidic bond connection mode of astragalus polysaccharides using GC-MS / MS detection technology and TG-1MS capillary column as the platform. The results are as follows: Figure 4 shown.

[0052] The characteristic ion pairs of m / z 129>87 and m / z 161 / 43 indicated that the peptide contained T-Araf. The characteristic ion pairs of m / z 117 / 43 and m / z 101 / 39 indicated that the peptide contained 4-Arap. The characteristic ion pairs of m / z 129 / 87 and m / z 161 / 43 indicated that the peptide contained T-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 126 / 82 indicated that the peptide contained 2-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 101 / 68 indicated that the peptide contained 3-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 113 / 41 indicated that the peptide contained 4-Glcp. The characteristic ion pair of m / z 99 / 39 can be used to determine that the compound contains a 6-Glcp connection mode; the characteristic ion pair of m / z 129 / 87 and m / z 131 / 41 can be used to determine that the compound contains a 2,4-Glcp connection mode; the characteristic ion pair of m / z 129 / 87 and m / z 169 / 109 can be used to determine that the compound contains a 3,4-Glcp connection mode; the characteristic ion pair of m / z 129 / 87 and m / z 186 / 84 can be used to determine that the compound contains a 3,6-Glcp connection mode; the characteristic ion pair of m / z 127 / 85 and m / z 85 / 55 can be used to determine that the compound contains a 4,6-Glcp connection mode; the characteristic ion pair of m / z 129 / 87 and m / z 197 / 112 can be used to determine that the compound contains a 2,3,4-Glcp connection mode; the characteristic ion pairs of m / z 127 / 85, m / z The characteristic ion pair of 184 / 66 can be determined to contain 3,4,6-Glcp connection mode.

[0053] Among them, the OCE value of 4-Xylp in Astragalus polysaccharide is 16.35eV, the OCE value of 4-Glcp is 21.04eV, the OCE value of 2,4-Glcp is 21.45eV, and the OCE value of 4,6-Glcp is 15.93eV. By comparing the values ​​in the OCE data array of partially methylated sugar nitrile acetate glycoside connection analysis, it can be confirmed that the above analysis results are correct.

[0054] The results of qualitative and quantitative analysis of the glycosidic bond connection modes of astragalus polysaccharides showed that the main glycosidic bond connection mode of astragalus polysaccharides was Glc, accounting for 93.90% of the polysaccharide content. Among them, T-Glcp, 4-Glcp and 3,4,6-Glcp were the main connection modes, and their percentages in the polysaccharides were 61.07%, 13.17% and 10.19% respectively.

[0055] Experimental Example 3 Application of the MRM data array and OCE data array constructed in Example 1 in the analysis of glycosidic bond connection mode of American ginseng polysaccharides

[0056] In order to accurately analyze the connection mode of glycosidic bonds in American ginseng polysaccharides, this experiment first detected American ginseng polysaccharides using the monosaccharide composition analysis method. The results showed that Glc was the main component of American ginseng polysaccharides. On this basis, the constructed partially methylated sugar nitrile acetate connection analysis qualitative and quantitative targeted MRM data array and OCE data array were applied to the analysis of glycosidic bond connection mode of American ginseng polysaccharides using GC-MS / MS detection technology and TG-1MS capillary column as a platform. The results are as follows: Figure 5 shown.

[0057] The characteristic ion pairs of m / z 129 / 87 and m / z 161 / 43 indicated that the molecule contained T-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 126 / 82 indicated that the molecule contained 2-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 101 / 68 indicated that the molecule contained 3-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 113 / 41 indicated that the molecule contained 4-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 99 / 39 indicated that the molecule contained 6-Glcp. The characteristic ion pairs of m / z 129 / 87 and m / z 131 / 41 indicated that the molecule contained 2,4-Glcp. The characteristic ion pair of 169 / 109 can be used to determine the presence of 3,4-Glcp connection mode; the characteristic ion pairs of m / z 129 / 87 and m / z 186 / 84 can be used to determine the presence of 3,6-Glcp connection mode; the characteristic ion pairs of m / z 127 / 85 and m / z 85 / 55 can be used to determine the presence of 4,6-Glcp connection mode; the characteristic ion pairs of m / z 129 / 87 and m / z 197 / 112 can be used to determine the presence of 2,3,4-Glcp connection mode.

[0058] Among them, the OCE value of 4-Glcp in American ginseng polysaccharide is 21.54eV, the OCE value of 2,4-Glcp is 21.75eV, and the OCE value of 4,6-Glcp is 15.53eV. By comparing the values ​​in the OCE data array of partially methylated sugar nitrile acetate glycoside connection analysis, it can be confirmed that the above analysis results are correct.

[0059] The results of qualitative and quantitative analysis of the glycosidic bond connection mode of American ginseng polysaccharides showed that the main glycosidic bond connection mode of American ginseng polysaccharides was Glc, among which T-Glcp, 4-Glcp and 4,6-Glcp were the main connection modes, and their percentages in the polysaccharides were 36.89%, 39.79% and 12.01%, respectively.

Claims

1. A qualitative analysis method for the monosaccharide glycosidic bond connection mode of traditional Chinese medicine polysaccharides, characterized in that: include: (1) preparing partially methylated nitrile acetates of different types of monosaccharides; (2) using GC-MS / MS method to separate different partially methylated sugar nitrile acetates and obtain EI-MS / MS spectra; (3) determining the partially methylated sugar nitrile acetate qualitative ion pair and the partially methylated sugar nitrile acetate quantitative ion pair according to the EI-MS / MS spectrum and constructing a targeted qualitative and quantitative multiple reaction monitoring data array in combination with the collision energy corresponding to the partially methylated sugar nitrile acetate qualitative ion pair and the partially methylated sugar nitrile acetate quantitative ion pair; (4) Calculating the optimal collision energy of partially methylated sugar nitrile acetate ion pairs to obtain the optimal collision energy data array; (5) Qualitative analysis of the glycosidic bond connection mode of traditional Chinese medicine polysaccharides based on multiple reaction monitoring data array or optimal collision energy data array; In step (3), according to the EI-MS / MS spectrum, an ion pair with a large response intensity is selected as a partially methylated sugar nitrile acetate quantitative ion pair, and an ion pair with structural specificity is selected as a partially methylated sugar nitrile acetate quantitative ion pair; wherein, under the background of a collision energy range of 5-45eV and a variation interval of 2eV, the response intensity of the ion pair is obtained, and the collision energy corresponding to the partially methylated sugar nitrile acetate qualitative ion pair and the partially methylated sugar nitrile acetate quantitative ion pair is determined by comparing the magnitude of the response intensity under different collision energies; The method for constructing the optimal collision energy data array by calculating the optimal collision energy value of the partially methylated sugar nitrile acetate ion pair in step (4) includes: obtaining the response intensity of the ion pair under the background of a collision energy range of 5-45eV and a variation interval of 2eV, fitting the obtained data with a normal regression curve through a Guass function, and calculating the optimal collision energy value of the partially methylated sugar nitrile acetate ion pair.

2. The qualitative analysis method according to claim 1, characterized in that: The different types of monosaccharides described in step (1) include, but are not limited to, any one or more of glucose, mannose, galactose, arabinose, D-xylose, ribose, rhamnose or fucose.

3. The qualitative analysis method according to claim 1, characterized in that: There are 24 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from glucose, 23 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from galactose, 12 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from arabinose, 12 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from D-xylose, 12 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from ribose, 8 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from rhamnose, and 8 glycosidic bond connection modes for the partially methylated sugar nitrile acetates obtained from fucose.

4. The qualitative analysis method according to claim 1, characterized in that: The parameters of the gas phase conditions and mass spectrometry conditions of the GC-MS / MS method in step (2) include: The gas phase conditions included: injection temperature of 250°C; injection volume of 1 μL; split ratio of 20:1, capillary column of TG-1MS, and heating program as follows: 110°C for 2 min, gradually heated to 210°C at a rate of 5°C / min and maintained for 2 min, then heated to 280°C at a rate of 20°C / min and maintained for 2 min; The mass spectrometry conditions included: EI source, ion source temperature of 280°C, scan rate of 0.2 scan / s, and automatic full scan mode with a scan range of m / z 50-550.

5. The qualitative analysis method according to claim 1, characterized in that: In step (5), the main components of the monosaccharides of the Chinese medicine polysaccharide to be detected are first analyzed, and then the monosaccharide glycosidic bond connection mode of the Chinese medicine polysaccharide is qualitatively analyzed based on the optimal collision energy data array.

6. The qualitative analysis method according to claim 1, characterized in that: In step (5), the isomers of the same monosaccharide are qualitatively analyzed based on the multiple reaction monitoring data array, or the isomers of some of the same monosaccharides and heterogeneous monosaccharides are qualitatively analyzed based on the optimal collision energy data array.

7. The qualitative analysis method according to claim 1, characterized in that: The traditional Chinese medicine polysaccharide is selected from any one of fungus, astragalus and American ginseng polysaccharides.

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