Polygonatum sibiricum medicinal material species identification method based on polygonatum sibiricum characteristic compound and application

Abstract

The invention discloses a polygonatum sibiricum medicinal material species identification method based on a polygonatum sibiricum characteristic compound and application of the polygonatum sibiricum medicinal material species identification method. The invention discloses a polygonatum sibiricum medicinal material species identification method based on a polygonatum sibiricum characteristic compound. The method comprises the following steps: 1) preparing a polygonatum sibiricum rhizome sample solution; 2) analyzing different sealwort rhizome metabolites by liquid chromatography-mass spectrometry; and 3) discriminating the species of the rhizoma polygonati medicinal material, namely discriminating the species of the rhizoma polygonati medicinal material if the rhizoma polygonati rhizome metabolite contains a corresponding characteristic compound. The polygonatum sibiricum variety is polygonatum kingianum, polygonatum sibiricum, polygonatum cyrtonema and polygonatum odoratum. According to the application of the polygonatum kingianum characteristic compounds in polygonatum kingianum medicinal material species identification, each polygonatum kingianum species selects three representative characteristic compounds which are relatively high in content, have functional specificity representatives and are common in the market, the species of the polygonatum kingianum medicinal material can be identified as long as the three representative characteristic compounds are contained at the same time, the accuracy is high, the number of components to be detected is small, and the accuracy is high. The detection period is short.

Description

Technical Field

[0001] The present invention belongs to the technical field of identification of medicinal herb species, and particularly relates to a method and application for identifying the species of Polygonatum sibiricum Redoute based on characteristic compounds of Polygonatum sibiricum Redoute. Background Art

[0002] Polygonatum sibiricum Redoute belongs to the genus Polygonatum of the family Liliaceae, and is a commonly used plant medicine, which has the effects of improving qi deficiency of the spleen and stomach, fatigue, yin deficiency of the stomach, dry mouth and less food intake, dry cough due to lung deficiency, cough with hemoptysis due to overstrain, insufficiency of essence and blood, soreness and weakness of the waist and knees, early whitening of hair and beard, and internal heat with thirst. Polygonatum sibiricum Redoute is included in the Chinese medicine and food homology catalog. The Polygonatum plants specified in the Chinese Pharmacopoeia (2020 Edition) include: Polygonatum kingianum Coll. et Hemsl. Polygonatum kingianum Coll. et Hemsl. ( Polygonatum sibiricum Red. ) Polygonatum cyrtonema Hua Polygonatum cyrtonema Hua ) Polygonatum cyrtonema Hua Polygonatum odoratum (Mill.) Druce). The rhizome morphologies of different species of Polygonatum sibiricum Redoute are relatively similar, and it is impossible to accurately distinguish them with the naked eye, making it difficult to identify the species of Polygonatum sibiricum Redoute. This restricts the development of the relevant Chinese patent medicine industry and is also a bottleneck problem encountered in the process of the modernization of traditional Chinese medicine in recent years.

[0003] In the Chinese Pharmacopoeia (2020 Edition), the method of observing the morphological characteristics of epidermal cells and vascular bundles in the cross-section is used to analyze and identify the species of Polygonatum sibiricum Redoute. In the prior art, there are methods for identifying the species of Polygonatum sibiricum Redoute based on gene markers. For example, in the relevant research of Guizhou University, a rapid, accurate and standardized method for identifying the species of Polygonatum sibiricum Redoute by DNA molecular markers based on the IRAP markers of Polygonatum sibiricum Redoute with retrotransposon sequences was established. Among them, the method for identifying the species of Polygonatum sibiricum Redoute by observing the morphological characteristics of epidermal cells and vascular bundles in the cross-section is greatly affected by the growth environment, harvesting time and growth years of Polygonatum sibiricum Redoute, lacking specificity and exclusivity, resulting in unstable accuracy of the identification of the species of Polygonatum sibiricum Redoute. Although the method for identifying the species of Polygonatum sibiricum Redoute based on DNA molecular markers is accurate, during the processes of washing, drying, storage, etc. of the medicinal materials, it will cause serious degradation of DNA, and will also be contaminated by exogenous DNA caused by microorganisms, all of which will make the extracted DNA have a low content and poor quality, making it difficult to carry out subsequent molecular identification and resulting in a relatively high operation difficulty.

[0004] Therefore, there is an urgent need in the prior art for a method and application for identifying the species of Polygonatum sibiricum Redoute based on characteristic compounds of Polygonatum sibiricum Redoute, which has strong specificity, high accuracy and simple operation. Summary of the Invention

[0005] The first object of the present invention is to overcome the deficiencies existing in the prior art and provide a method for identifying the species of Polygonatum sibiricum Redoute based on characteristic compounds of Polygonatum sibiricum Redoute, which has strong specificity, high accuracy and simple operation.

[0006] The first object of the present invention is achieved by the following technical solutions: A method for identifying the species of Polygonatum odoratum medicinal materials based on characteristic compounds of Polygonatum odoratum, comprising the following steps: 1) Preparation of a solution of Polygonatum odoratum rhizome samples; 2) Liquid chromatography-mass spectrometry analysis of metabolites in different Polygonatum odoratum rhizomes; 3) Discrimination of the species of Polygonatum odoratum medicinal materials: If the metabolites in the Polygonatum odoratum rhizome contain corresponding characteristic compounds, the species of the Polygonatum odoratum medicinal materials can be identified; The Polygonatum odoratum varieties are Polygonatum kingianum, Polygonatum cyrtonema, Polygonatum corymbiflorum and Polygonatum odoratum; The characteristic compounds of Polygonatum kingianum are as follows: Number Compound Name Molecular Formula CAS No. 1 Pizotyline C19H21NS 15574-96-6 2 Imidapril C20H27N3O6 89371-37-9 3 3-Polyprenyl-4,5-dihydroxybenzoate C17H22O4 4 6-{3,5-dihydroxy-2-[3-(3,4,5-trimethoxyphenyl)propanoyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid C24H28O13 5 1-Guanidino-1-deoxy-scyllo-inositol 4-phosphate C7H16N3O8P 6 Isocyclomorusin C25H22O6 7 Pyracarbolid C13H15NO2 24691-76-7 8 Virginiamycin M1 C28H35N3O7 21411-53-0 9 5'-Butyrylphosphouridine C13H19N2O10P 10 Compound IV C20H26N2O3 87495-31-6 11 PKOOA-PI C33H57O15P 12 Bisindolylmaleimide IV C20H13N3O2 119139-23-0 13 PA(15:0 / 14:1(9Z)) C32H61O8P 14 Estrone C18H22O2 53-16-7 15 Dextromoramide C25H32N2O2 5666-11-5 16 Galabiose C24H44O22 17 1-Dehydro-9-fluoro-11-oxotestololactone C19H21FO4 6875-24-7 18 Leurosine C46H56N4O9 23360-92-1 19 18-Fluoro-9,10-Epoxyoctadecanoic acid C18H33FO3 20 Cannabidiolic acid C22H30O4 1244-58-2 21 Lyciumoside VII C44H74O21 22 1,2,3,4-Tetrahydro-2-[(isopropylamino)methyl]-7-nitro-6-quinolinecarboxylic acid C14H19N3O4 23 PtdIns-(4,5)-P2 (1,2-dipalmitoyl) (sodium salt) C41H81O19P3 24 PS(16:0 / 18:2(9Z,12Z)) C40H74NaO10P 322647-44-9 25 xi-Linalool 3-[rhamnosyl-(1->6)-glucoside] C22H38O10 26 6'-HMG SDG C38H54O20 27 Tetrahydrogeranylgeranyl diphosphate C20H40O7P2 28 Isoleucyl-Threonine C10H20N2O4 29 PA(18:3(9Z,12Z,15Z) / 20:5(5Z,8Z,11Z,14Z,17Z)) C41H65O8P 30 1,3-(8R,9R-epoxy-octadec-13Z,15Z-dien-4,6-diynoyl)-2-(8-hydroxy-13E,17E-octadecadien-9,11-diynoyl)-sn-glycerol C56H68O9 31 Granisetron metabolite 1 C18H24N4O3 32 CL(8:0 / 8:0 / 8:0 / 11:0) C44H84O17P2 33 PA(20:3(5Z,8Z,11Z) / 15:0) C38H69O8P 34 PI(22:6(4Z,7Z,10Z,13Z,16Z,19Z) / 18:4(6Z,9Z,12Z,15Z)) C49H75O13P 35 N-Formyl-norleucyl-leucyl-phenylalanyl-methylester C23H35N3O5 36 Benzhydrylpiperazine C26H28N2O The characteristic compounds of Polygonatum cyrtonema are as follows: Number Compound Name Molecular Formula CAS No. 1 2-Hydroxy-1,4-benzoquinone C6H4O3 2474-72-8 2 Xanthoplanine C21H26NO4 6872-88-4 3 N-Carboxytocainide C12H16N2O3 4 Sulfamerazine C11H12N4O2S 127-79-7 5 L-902,688 C21H27F2N5O2 634193-54-7 6 7-Methylxanthosine C11H14N4O6 29885-96-9 7 Niazidin C15H18N2O6S 8 LysoPE(0:0 / 16:1(9Z)) C21H42NO7P 9 Tinidazole C8H13N3O4S 19387-91-8 10 Carboxy-ibuprofen C13H16O4 11 Rivenprost C24H34O6S 256382-08-8 12 Gallic acid C7H6O5 149-91-7 13 N-(2E,4E,10E,12Z-tetradecatetraen-8-yn-oyl) isobutylamine C18H25NO 14 Angelicolide C24H28O4 90826-58-7 15 Dialicor C21H27ClNO2 2192-21-4 16 2,3-Diphenyl-3-(2-pyridinyl)acrylonitrile C20H14N2 39077-64-0 17 Sphingosine-1-phosphate C18H38NO5P 26993-30-6 18 Lophophorine C13H17NO3 17627-78-0 19 Isocyclocalamin C27H34O9 20 PC(O-12:0 / O-1:0) C21H46NO6P 21 2-mercapto-octadecanoic acid C18H36O2S 600730-11-8 22 Verruculogen C27H33N3O7 23 Gossypetin 7-methyl ether 8-acetate C18H14O9 24 PS(19:1(9Z) / 0:0) C25H48NO9P 25 PI(P-16:0 / 12:0) C37H71O12P 26 Narasin B C43H70O11 The characteristic compounds of Polygonatum corymbiflorum are as follows: Number Compound Name Molecular Formula CAS No. 1 L-Histidinol phosphate C6H12N3O4P 25679-93-0 2 Alternariol C14H10O5 641-38-3 3 Metamitron C10H10N4O 41394-05-2 4 Nicotinuric acid C8H8N2O3 583-08-4 5 2-Decarboxybetanin C23H27N2O11+ 6 7-Methylthioheptyl glucosinolate C15H29NO9S3 7 Amylopectin C30H52O26 9037-22-3 8 Torachrysone 8-(6-oxalylglucoside) C22H24O12 9 Bluensomycin C21H39N5O14 11011-72-6 10 PC(16:0 / 25:0)[U] C49H98NO8P The characteristic compounds of Polygonatum odoratum are as follows: Number Compound Name Molecular Formula CAS No. 1 1-(Methylsulfinyl)propyl propyl disulfide C7H16OS3 188055-44-9 2 PA(20:4(5Z,8Z,11Z,14Z) / 20:5(5Z,8Z,11Z,14Z,17Z)) C43H67O8P 474943-33-4 3 Olomoucine C15H18N6O 101622-51-9 4 BRUCINE C23H26N2O4 5892-11-5 5 PE(20:4(5Z,8Z,11Z,14Z) / 0:0) C25H44NO7P In step 3), if the metabolites in the Polygonatum odoratum rhizome contain more than 3 corresponding characteristic compounds, the species of the Polygonatum odoratum medicinal materials can be identified.

[0007] In step 3), if the metabolites in the Polygonatum odoratum rhizome contain Estrone, Leurosine and Cannabidiolic acid, the species of the Polygonatum odoratum medicinal materials can be identified as Polygonatum kingianum; If the metabolites in the Polygonatum odoratum rhizome contain Xanthoplanine, Gallic acid and Sphingosine-1-phosphate, the species of the Polygonatum odoratum medicinal materials can be identified as Polygonatum cyrtonema; If the metabolites in the Polygonatum odoratum rhizome contain Alternariol, Nicotinuric acid and Amylopectin, the species of the Polygonatum odoratum medicinal materials can be identified as Polygonatum corymbiflorum; If the metabolites in the Polygonatum odoratum rhizome contain 1-(Methylsulfinyl)propyl propyl disulfide, Olomoucine and BRUCINE, the species of the Polygonatum odoratum medicinal materials can be identified as Polygonatum odoratum.

[0008] The second object of the present invention is to overcome the deficiencies of the existing technology and provide the application of characteristic compounds of Polygonatum odoratum in the identification of the species of Polygonatum odoratum medicinal materials.

[0009] The second object of the present invention is achieved through the following technical solutions: the application of the characteristic compounds of Polygonatum in the species identification of Polygonatum medicinal materials, by analyzing the metabolite sample solution of Polygonatum rhizome through liquid chromatography-mass spectrometry; Estrone, Leurosine and Cannabidiolic acid are contained in the metabolites of Polygonatum rhizome, and it can be identified that the species of this Polygonatum medicinal material is Polygonatum kingianum Coll. et Hemsl.; Xanthoplanine, Gallic acid and Sphingosine-1-phosphate are contained in the metabolites of Polygonatum rhizome, and it can be identified that the species of this Polygonatum medicinal material is Polygonatum sibiricum Red.; Alternariol, Nicotinuric acid and Amylopectin are contained in the metabolites of Polygonatum rhizome, and it can be identified that the species of this Polygonatum medicinal material is Polygonatum cyrtonema Hua; 1-(Methylsulfinyl)propyl propyl disulfide, Olomoucine and BRUCINE are contained in the metabolites of Polygonatum rhizome, and it can be identified that the species of this Polygonatum medicinal material is Polygonatum odoratum (Mill.) Druce.

[0010] The beneficial effects of the present invention are as follows: by analyzing the secondary metabolite compounds in the rhizomes of 4 Polygonatum species, 36 characteristic compounds are found in Polygonatum kingianum Coll. et Hemsl., 26 characteristic compounds are found in Polygonatum sibiricum Red., 10 characteristic compounds are found in Polygonatum cyrtonema Hua, and 5 characteristic compounds are found in Polygonatum odoratum (Mill.) Druce. For each Polygonatum species, 3 representative characteristic compounds with relatively high content, functional specificity and common in the market are selected. When detected by the LC-ESI-Q-TOF method, as long as the corresponding three representative characteristic compounds are contained at the same time, the species of this Polygonatum medicinal material can be identified, with high accuracy, few components to be detected and short detection cycle. Description of the Drawings

[0011] Figure 1 It is the total ion current chromatogram of the metabolites of Polygonatum cyrtonema Hua; Figure 2 It is the total ion current chromatogram of the metabolites of Polygonatum kingianum Coll. et Hemsl.; Figure 3 It is the total ion current chromatogram of the metabolites of Polygonatum odoratum (Mill.) Druce; Figure 4 It is the total ion current chromatogram of the metabolites of Polygonatum sibiricum Red.; Figure 5Principal component analysis chart of the rhizome metabolites of four Polygonatum plants, where the abscissa PC1 represents the first principal component, the ordinate PC2 represents the second principal component, QC represents quality control, and PK represents Polygonatum kingianum Polygonatum kingianum (Coll. et Hemsl.), PS represents Polygonatum sibiricum Polygonatum sibiricum (Red.), PC represents Polygonatum cyrtonema Polygonatum cyrtonema (Hua), and PO represents Polygonatum odoratum Polygonatum odoratum (Mill.) Druce). Specific implementation mode

[0012] The present invention will be described in detail below with reference to the accompanying drawings.

[0013] Example Analysis of characteristic compounds of Polygonatum plants S1. Preparation of sample solution: Collect fresh rhizomes of Polygonatum kingianum Polygonatum kingianum Coll. et Hemsl. (), Polygonatum sibiricum Polygonatum sibiricum Red. (), Polygonatum cyrtonema Polygonatum cyrtonema Hua (), and Polygonatum odoratum Polygonatum odoratum (Mill.) Druce () at the mature stage. After cryogenic freeze-drying, grind them into powder, and then operate according to the following steps: 1. Weigh 80 mg of the sample, add 20 μL of the internal standard solution (L-2-chlorophenylalanine, 0.3 mg / mL; prepared with methanol) and 1 mL of methanol-water (V:V = 7:3); 2. Add two small steel balls, place them at -20 °C for 2 min for pre-cooling, and add them to the grinder (60 Hz, 2 min); 3. Ultrasonic extract in an ice-water bath for 30 min and let it stand overnight at -20 °C; 4. Centrifuge for 10 min (13000 rpm, 4 °C), suck 150 μL of the supernatant with a syringe, filter it through a 0.22 μm organic phase needle filter, transfer it to an LC injection vial, and store it at -80 °C until LC-MS analysis; 5. The quality control sample (QC) is prepared by mixing equal volumes of the extraction solutions of all samples, and the volume of QC is the same as that of the samples.

[0014] All of the above extraction reagents are pre-cooled at -20 °C before use.

[0015] S2. A liquid chromatography - mass spectrometry system composed of a Dionex U3000 UHPLC ultra - high performance liquid chromatography tandem QE high - resolution mass spectrometer was used to analyze the rhizome metabolites of different species of Polygonatum: The sample solution was subjected to ultra - high performance liquid chromatography tandem mass spectrometry analysis. An ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 um) was used; the column temperature was 45°C; mobile phase A was ultrapure water containing 0.1% (V / V) formic acid, and mobile phase B was acetonitrile containing 0.1% (V / V) formic acid; the gradient elution program was as follows: at 0.01 min, the proportion of phase B was 5%, within 2.00 min, the proportion of phase B was 5%, within 4.00 min, the proportion of phase B was 30%, within 8.00 min, the proportion of phase B was 50%, within 10.00 min, the proportion of phase B was 80%, within 14.00 min, the proportion of phase B was 100%, within 15.00 min, the proportion of phase B was 100%, at 15.10 min, the proportion of phase B was 5%, within 18.00 min, the proportion of phase B was 5%; the flow rate was 0.35 mL / min; the column temperature was 40°C; the injection volume was 2 μL; Mass spectrometry conditions: The ion source was ESI; the sample mass spectrometry signal acquisition was performed using positive and negative ion scanning modes respectively; the ESI ion source temperature was 350°C; the mass - to - charge ratio range of the mass spectrometer was 100 to 1000. The resolution of the full mass spectrometry scan was set to 70,000, and the resolution of the MS / MS scan was set to 17,500; the collision energy was set to 10, 20, and 40 electron volts.

[0016] The mass spectrometer was operated as follows: The ion spray voltage was 3800 V (positive ion mode) / - 3200 V (negative ion mode); the sheath gas flow rate was 40 units; the auxiliary gas flow rate was 8 units; the capillary temperature was 320°C. During the analysis, QCs were injected at fixed intervals for every 10 samples to provide a set of data that could be used to evaluate the repeatability.

[0017] S3. Qualitative and quantitative analysis of Polygonatum metabolites: Based on the non - targeted metabolomics of ultra - high performance liquid chromatography tandem high - resolution mass spectrometry, combined with the metabolomics data processing software Progenesis QI v2.3, qualitative and relative quantitative analysis of the raw data was carried out, and the raw data was pre - processed by standardization.

[0018] The BPC (Base Peak Chromatogram) is a chromatogram obtained by continuously depicting the intensity of the strongest ion in the mass spectrometry diagram at each time point. As Figures 1 - 4 are the total ion current diagrams of the positive and negative ion modes of 4 species of Polygonatum plants. The components eluted through chromatographic separation continuously enter the mass spectrometer, and the mass spectrometer continuously scans for data acquisition. Each scan obtains a mass spectrometry diagram. The ion with the largest intensity in each mass spectrometry diagram is continuously depicted. With the ion intensity as the vertical coordinate and time as the horizontal coordinate, the resultingFigures 1 - 4 Chromatogram. The results showed that the peak emergence times of the BPC chromatograms of the 4 samples were basically the same, and the corresponding intensities and retention times of the chromatographic peaks were basically overlapped. The variation caused by instrument error during the whole experimental process was small, and the data quality was reliable, indicating that the 4 kinds of Polygonatum sibiricum Delar. ex Redoute contained most of the same metabolites. In addition, there were differences in the intensities and numbers of some chromatographic peaks, indicating that there were differences in the types and contents of metabolites among the 4 kinds of Polygonatum sibiricum Delar. ex Redoute.

[0019] S4, Quality control: Through the quality control of QC samples, the pretreatment of the experimental part, sample injection into the instrument, and the stability of the mass spectrometry system of this project were analyzed and evaluated.

[0020] QC sample quality control: Before sample detection, QC samples were used to balance the "chromatography - mass spectrometry" system, and during sample detection, they were used to evaluate the stability of the mass spectrometry system.

[0021] (1) RSD screening of QC samples Delete the ion peaks with RSD > 0.4 in the QC group. Relative standard deviation (RSD) is the ratio of the standard deviation to the arithmetic mean of the measurement results, a standard for measuring the dispersion degree of data distribution, used to measure the degree to which data values deviate from the arithmetic mean. The smaller the standard deviation, the less these values deviate from the mean, and vice versa.

[0022] (2) Principal component analysis of QC samples Principal Component Analysis is an unsupervised data analysis method. It uses an orthogonal transformation to transform the original random vector with correlated components into a new random vector with uncorrelated components, so that they can reflect as much of the original variable information as possible, thereby achieving the purpose of dimensionality reduction. At the same time, the system stability was evaluated. As Figure 5 shown, in the PCA model diagram obtained by 7 - fold cross - validation, the QC samples were closely clustered together, indicating that the instrument detection stability was good during this experimental process. Principal component analysis was used to characterize the overall metabolic differences of the rhizomes of four Polygonatum plants. The results showed that the rhizome samples of the four Polygonatum plants were distributed in different regions of the graph, indicating that the metabolites of different varieties were different.

[0023] S5. Screening of differential metabolites: A combination of multidimensional analysis and unidimensional analysis was used to screen differential metabolites between groups. In OPLS-DA analysis, variable weights (VIP) were used to measure the influence and explanatory power of the expression patterns of each metabolite on the classification and discrimination of each group of samples, to mine differential metabolites with biological significance, and to further use the T test to verify whether the differential metabolites between groups were significant. The screening criteria were OPLS-DA model, VIP value of the first principal component>1, and p-value of the T test<0.05. The specific screening methods for differential metabolites are shown in Table 1.

[0024] Table 1 Differential metabolite screening table Parameter Parameter Explanation VIP Variable importance in projection, the VIP value from the OPLS-DA model. The larger the VIP, the greater the contribution of the variable to the grouping. P-value The results of the t-test are used to evaluate whether the differences in variables between two groups of samples are significant. A p < 0.05 indicates significance adj.P-value False discovery rate (FDR), used to control the proportion of false positive results in differential analysis, calculated by the Benjamini-Hochberg method Log2(FC) The ratio of the average expression levels of metabolites in two groups of samples. A positive value indicates upregulation, and a negative value indicates downregulation. Average(#) Average expression level of metabolites in group # LC-ESI-Q-TOF method was used to analyze the Polygonatum kingianum Coll. et Hemsl. )、Acanthopanax odoratus( Polygonatum sibiricum Red. )、Potamogeton cymbidium( Polygonatum cyrtonema Hua ) and Polygonatum odoratum ( Polygonatum odoratum (Mill.) Druce) and the results showed that 2381, 2360, 2355 and 2336 unique metabolites were detected, respectively, and there was considerable overlap in the metabolite profiles detected in the four species. More importantly, 36 unique compounds were found in Polygonatum kingianum Coll. et Hemsl. (Table 2), 26 unique compounds were found in Polygonatum sibiricum Red. (Table 3), 10 unique compounds were found in Polygonatum cyrtonema Hua (Table 4), and 5 unique compounds were found in Polygonatum odoratum (Mill.) Druce (Table 5). The specific metabolites of these four plants can serve as clues to distinguish different varieties and provide targets for future experiments.

[0025] Table 2

[0026] In Yunnan Polygonatum Polygonatum kingianumColl. et Hemsl.), Polygonatum cirrhifolium (Wall.) Royle), and Polygonatum odoratum (Mill.) Druce), see Table 6, three representative characteristic compounds with relatively high contents, functional specificity, and commonness in the market were selected for each Polygonatum species. According to the data analysis results of specific metabolites and the above selection requirements, Estrone, Leurosine, and Cannabidiolic acid were used as the representative characteristic compounds of Polygonatum kingianum; Xanthoplanine, Gallic acid, and Sphingosine-1-phosphate (D-threo-sphingosine-1-phosphate) were used as the representative characteristic compounds of Polygonatum cirrhifolium; Alternariol, Nicotinuric acid, and Amylopectin were used as the representative characteristic compounds of Polygonatum cyrtonema; 1-(Methylsulfinyl)propyl propyl disulfide, Olomoucine (2-(2'-hydroxyethylamino)-9-methyl-6-(benzylamino)purine), and BRUCINE were used as the representative characteristic compounds of Polygonatum odoratum. Polygonatum sibiricum Red.), Polygonatum cyrtonema Hua), and Polygonatum odoratum Polygonatum cyrtonema Hua) and Polygonatum odoratum Polygonatum odoratum (Mill.) Druce) among the above characteristic compounds detected in the rhizomes of Polygonatum kingianum Coll. et Hemsl., Polygonatum cirrhifolium (Wall.) Royle), Polygonatum cyrtonema Hua, and Polygonatum odoratum (Mill.) Druce), see Table 6, three representative characteristic compounds with relatively high contents, functional specificity, and commonness in the market were selected for each Polygonatum species. According to the data analysis results of specific metabolites and the above selection requirements, Estrone, Leurosine, and Cannabidiolic acid were used as the representative characteristic compounds of Polygonatum kingianum; Xanthoplanine, Gallic acid, and Sphingosine-1-phosphate (D-threo-sphingosine-1-phosphate) were used as the representative characteristic compounds of Polygonatum cirrhifolium; Alternariol, Nicotinuric acid, and Amylopectin were used as the representative characteristic compounds of Polygonatum cyrtonema; 1-(Methylsulfinyl)propyl propyl disulfide, Olomoucine (2-(2'-hydroxyethylamino)-9-methyl-6-(benzylamino)purine), and BRUCINE were used as the representative characteristic compounds of Polygonatum odoratum.

[0027]

[0028] The three representative characteristic compounds can accurately identify the four Polygonatum species. When detected by the LC-ESI-Q-TOF method, as long as the corresponding three representative characteristic compounds are contained simultaneously, the species of the Polygonatum medicinal material can be distinguished, with high accuracy.

[0029] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than limiting the protection scope of the present invention. Any simple modification or equivalent replacement of the technical solution of the present invention by those of ordinary skill in the art shall not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A method for identifying the species of Polygonatum sibiricum based on characteristic compounds of Polygonatum sibiricum, characterized in that The steps include: 1) Preparation of Polygonatum cyrtonema rhizome sample solution; 2) Liquid chromatography-mass spectrometry analysis of metabolites of different Polygonatum rhizomes; 3) Identification of the species of Polygonatum sibiricum: The metabolites of Polygonatum sibiricum rhizomes contain corresponding characteristic compounds, which can be used to identify the species of the Polygonatum sibiricum; The varieties of polygonatum are polygonatum yunnanensis, polygonatum cyrtonema, polygonatum multiflorum and polygonatum odoratum; The characteristic compounds of the Yunnan Polygonatum are as follows: The characteristic compounds of the Polygonatum sibiricum are as follows: The characteristic compounds of Polygonatum cyrtonema are as follows: The characteristic compounds of the polygonatum odoratum are as follows: 。 2. The method for identifying the species of polygonatum sibiricum based on characteristic compounds of polygonatum sibiricum according to claim 1, characterized in that: In step 3), if the rhizome metabolites of Polygonatum sibiricum contain more than three corresponding characteristic compounds, the species of the Polygonatum sibiricum medicinal material can be identified.

3. The method for identifying the species of polygonatum sibiricum based on characteristic compounds of polygonatum sibiricum according to claim 2, characterized in that: In step 3), the metabolites of Polygonatum sibiricum rhizome contain Estrone, Leurosine and Cannabidiolic acid, which can identify the species of the Polygonatum sibiricum medicinal material as Yunnan Polygonatum sibiricum; The metabolites of Polygonatum sibiricum rhizome contain Xanthoplanine, Gallic acid and Sphingosine-1-phosphate, which can identify the species of Polygonatum sibiricum as Polygonatum cyrtonema; The metabolites of Polygonatum sibiricum rhizome contain Alternariol, Nicotinuric acid and Amylopectin, which can identify the species of Polygonatum sibiricum as Polygonatum cyrtonema. The metabolites of the rhizome of Polygonatum sibiricum contain 1-(Methylsulfinyl)propyl propyl disulfide, Olomoucine and BRUCINE, which can identify the species of the medicinal material of Polygonatum sibiricum as Polygonatum odoratum.

4. The application of characteristic compounds of polygonatum to species identification of polygonatum medicinal materials is characterized by: The sample solution of the metabolites of the rhizome of Polygonatum sibiricum was analyzed by liquid chromatography-mass spectrometry; the metabolites of the rhizome of Polygonatum sibiricum contained Estrone, Leurosine and Cannabidiolicacid, which could identify the species of the Polygonatum sibiricum as Polygonatum yunnanensis; the metabolites of the rhizome of Polygonatum sibiricum contained Xanthoplanine, Gallic acid and Sphingosine-1-phosphate, which could identify the species of the Polygonatum sibiricum as Polygonatum cyrtonema; the metabolites of the rhizome of Polygonatum sibiricum contained Alternariol, Nicotinuric acid and Amylopectin, which could identify the species of the Polygonatum sibiricum as Polygonatum cyrtonema; the metabolites of the rhizome of Polygonatum sibiricum contained 1-(Methylsulfinyl)propyl propyldisulfide, Olomoucine and BRUCINE, which could identify the species of the Polygonatum sibiricum as Polygonatum odoratum.

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