Method for identifying radix tetrastigme cultivated in wild-imitated mode and radix tetrastigme cultivated in greenhouse

Abstract

The invention discloses a method for identifying radix tetrastigme cultivated in a wild-imitating manner and radix tetrastigme cultivated in a greenhouse. The specific stable isotope ratio and the specific mineral element content which have significant differences in the radix tetrastigme cultivated in an imitated wild mode and the radix tetrastigme cultivated in a greenhouse are adopted, and particularly the specific stable isotope ratio and the specific mineral element content which have significant differences in different parts of the radix tetrastigme are adopted. Specific identification models of the leaves, the stems, the tuberous roots and the fibrous roots of the radix tetrastigme are established, the radix tetrastigme cultivated in a wild-imitating mode and the radix tetrastigme cultivated in a greenhouse can be accurately and efficiently identified from the leaves, the stems, the tuberous roots and the fibrous roots, and the accuracy rate is 100%.

Description

Technical Field

[0001] The invention relates to the technical field of Chinese medicinal material identification, and in particular to a method for identifying wild-simulated cultivated Tripterygium wilfordii and greenhouse-cultivated Tripterygium wilfordii. Background Art

[0002] Tetrastigma hemsleyanum Diels et Gilg belongs to the genus Tetrastigma of the grape family. It is also known as stone monkey, snake aconite, golden silk hanging gourd, etc. Its scientific name is Tetrastigma hemsleyanum. It is often used as medicine with tubers or whole plants. Tetrastigma hemsleyanum mainly contains active ingredients such as flavonoids, flavonoid glycosides, reducing sugars, steroid compounds, amino acids, etc. It has pharmacological activities such as accelerating cancer cell apoptosis, inhibiting cancer cell growth, and improving body immunity in clinical medicine. Wild Tetrastigma hemsleyanum usually grows in the undergrowth of the forest on the hillside at an altitude of 300m-1300m, and in the cracks of rocks under the forest beside the valley stream. With the increasing importance of Tetrastigma hemsleyanum in medicine, it has gradually entered people's field of vision, and the market demand has been continuously increasing. However, the resources of wild Tetrastigma hemsleyanum are limited. In order to protect the resources of Tetrastigma hemsleyanum and promote the development of the Tetrastigma hemsleyanum industry, the artificial cultivation industry of Tetrastigma hemsleyanum has been continuously developing. At present, there are two main artificial cultivation methods of Tetrastigma hemsleyanum: wild cultivation (simulated wild cultivation) and greenhouse cultivation (greenhouse cultivation). Compared with greenhouse-cultivated Tripterygium wilfordii, simulated wild-cultivated Tripterygium wilfordii has richer active substances such as polysaccharides, saponins, and amino acids. Therefore, the market price of simulated wild-cultivated Tripterygium wilfordii is higher than that of greenhouse-cultivated Tripterygium wilfordii, and people have a higher desire to consume simulated wild-cultivated Tripterygium wilfordii. Driven by profit, many unscrupulous vendors choose to mislabel Tripterygium wilfordii when selling it, and sell greenhouse-cultivated Tripterygium wilfordii at the price of simulated wild-cultivated Tripterygium wilfordii to obtain higher profits. Since simulated wild-cultivated Tripterygium wilfordii and greenhouse-cultivated Tripterygium wilfordii are the same kind of Chinese medicinal materials, it is difficult to distinguish them from the perspectives of appearance, fingerprint patterns, etc. Therefore, it is realistic to establish a method for identifying simulated wild-cultivated Tripterygium wilfordii and greenhouse-cultivated Tripterygium wilfordii.

[0003] In recent years, the identification technology of Tripterygium wilfordii has focused on the identification of the authenticity of Tripterygium wilfordii and the identification of the origin of Tripterygium wilfordii. Chinese patent application CN103421907A discloses a method for identifying the medicinal plant Tripterygium wilfordii. Through the specific nucleotide sequence of Tripterygium wilfordii and specific primers based on the sequence, PCR technology is used to quickly and accurately identify Tripterygium wilfordii, providing an objective, fast and accurate method for the identification of the medicinal plant Tripterygium wilfordii, and solving the problem of counterfeit Tripterygium wilfordii in the medicinal material market to deceive consumers. Chinese patent application CN107101972A discloses a method for rapid identification of the origin of Tripterygium wilfordii by near-infrared spectroscopy. Combined with the characteristics of Tripterygium wilfordii near-infrared spectral data, the algorithm has been improved in three aspects: ① Estimating the probability density of samples from the perspective of distance; ② Calculating the bandwidth parameter in the form of training sample credibility; ③ In the identification of unknown new origins, a recognition method based on the probability density function of training set samples is proposed. Chinese patent application CN112684096A discloses a method for identifying the medicinal material of Tripterygium wilfordii based on polyamide thin layer chromatography, which can complete the identification of flavonoids in Tripterygium wilfordii medicinal materials under normal pressure, which is convenient, fast, accurate and reliable, and provides technical support for the quality control of Tripterygium wilfordii drugs. Chinese patent application CN113466224A discloses an array sensor for identifying the origin of Tripterygium wilfordii and its preparation method and application, which innovatively applies colorimetric array sensing technology to the identification of the origin of Tripterygium wilfordii. The prepared array sensor is fast in identifying the origin of Tripterygium wilfordii, only needs 5-10min, is accurate, efficient and low in cost. However, these methods are not suitable for the identification of simulated wild cultivated Tripterygium wilfordii and greenhouse cultivated Tripterygium wilfordii, and there is currently no effective method for identifying simulated wild cultivated Tripterygium wilfordii and greenhouse cultivated Tripterygium wilfordii. Summary of the invention

[0004] In order to solve the problem that there is no effective identification method for simulated wild-cultivated Tripterygium wilfordii and greenhouse-cultivated Tripterygium wilfordii in the prior art, the present invention provides an accurate and efficient method for identifying simulated wild-cultivated Tripterygium wilfordii and greenhouse-cultivated Tripterygium wilfordii.

[0005] The present invention finds that there are significant differences in specific stable isotope ratios and specific mineral element contents between simulated wild-cultivated clover and greenhouse-cultivated clover, especially there are significant differences in specific stable isotope ratios and specific mineral element contents in different parts of clover. Based on the finding, the present invention establishes several groups of specific identification models, which can accurately and efficiently identify simulated wild-cultivated clover and greenhouse-cultivated clover.

[0006] A method for identifying simulated wild-cultivated clover and greenhouse-cultivated clover, comprising using one or more of leaves, stems, tuberous roots, and fibrous roots of a clover sample to be tested to identify simulated wild-cultivated clover and greenhouse-cultivated clover;

[0007] The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the leaves of the tested Tripterygium wilfordii sample comprises: determining the δ 15 The N stable isotope ratio, Pb element content and S element content were respectively substituted into the following identification model of the leaves of the three-leaf green for calculation, and the F 大棚叶 and F 仿野生叶 If F 大棚叶 >F 仿野生叶 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生叶 >F 大棚叶 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii;

[0008] F 大棚叶 =2.462δ 15 N+2.990Pb+0.022S-36.876

[0009] F 仿野生叶 =0.584δ 15 N+14.090Pb+0.016S-25.508;

[0010] The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the stem of the tested Tripterygium wilfordii sample comprises: determining the δ 13 C stable isotope ratio, δ 15 The N stable isotope ratio and Ca content were substituted into the following identification model of the stem of Tripterygium wilfordii for calculation, and the F 大棚茎 and F 仿野生茎 If F 大棚茎 >F 仿野生茎 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生茎 >F 大棚茎 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii;

[0011] F 大棚茎 =0.652δ 15 N+0.007Ca-21.506δ 13 C-359.658

[0012] F 仿野生茎 =-0.898δ 15 N+0.006Ca-22.990δ 13 C-390.142;

[0013] The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the tubers of the tested Tripterygium wilfordii sample comprises: determining the δ 13 C stable isotope ratio, δ 15The N stable isotope ratio and Na content were substituted into the following identification model of the root of Tripterygium wilfordii for calculation, and the F 大棚块根 and F 仿野生块根 If F 大棚块根 >F 仿野生块根 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生块根 >F 大棚块根 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii;

[0014] F 大棚块根 =4.599δ 15 N+0.051Na-19.317δ 13 C-297.410

[0015] F 仿野生块根 =3.942δ 15 N+0.044Na-19.926δ 13 C-311.255;

[0016] The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the fibrous roots of the tested Tripterygium wilfordii sample comprises: determining the δ 13 C stable isotope ratio, δ 15 The N stable isotope ratio and Ca element content were substituted into the following identification model of the root hairs of Tripterygium wilfordii for calculation, and the F 大棚须根 and F 仿野生须根 If F 大棚须根 >F 仿野生须根 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生须根 >F 大棚须根 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii;

[0017] F 大棚须根 =-9.164δ 15 N+0.010Ca-38.071δ 13 C-600.163

[0018] F 仿野生须根 =-11.746δ 15 N+0.010Ca-41.321δ 13 C-694.687;

[0019] The content of each element is expressed in mg / kg, and the stable isotope ratio is expressed in ‰.

[0020] The method for determining the ratio of each stable isotope and the content of each element in the sample to be tested of the present invention can select the existing detection method. In order to achieve better technical effects, the following optimization is performed:

[0021] The stable isotope ratio calculation formula is: δE=(R 样品 / R 标准 -1);

[0022] Where, E is the target element isotope; R 样品 is the abundance ratio of heavy isotopes to light isotopes in the measured sample, that is, 13 C / 12 C. 15 N / 14 N, R 标准 is the abundance ratio of heavy isotopes to light isotopes in the standard sample; the δ value is multiplied by 1000 to obtain the final unit ‰, and the result is calibrated using the international unified standard unit, δ 13 C uses the natural abundance relative to the VPDB standard, δ 15 N refers to the abundance of nitrogen relative to air.

[0023] The isotope abundance in each sample can be determined by elemental analysis-stable isotope ratio mass spectrometry. 13 C.δ 15 The detection conditions for elemental analysis during N determination are preferably: the main filler of the combustion tube is WO 3 , 1150℃; reduction tube filler is Cu, 850℃; helium is used as carrier gas; the detection conditions of stable isotope ratio mass spectrometry are: CO 2 Sink current 100μA, N 2 Sink current 400μA.

[0024] The sample of the three-leaf green to be tested is first pretreated and then the content of each element is determined. The pretreatment includes: classifying the sample of the three-leaf green to be tested according to leaves, stems, tubers, and fibrous roots and drying them, crushing them and passing them through a 100-mesh sieve, collecting the sieve under the sieve and placing them in a high-pressure digestion tank, adding nitric acid, and after the sample is fully infiltrated by the acid solution, placing it in a 120°C pre-digestion until the yellow smoke dissipates, cooling it and sealing it, placing it in a microwave digestion instrument for digestion, and the digestion conditions are: first heating to 120°C for 5 minutes, keeping the temperature at 120°C for 5 minutes, then heating to 160°C for 5 minutes, keeping the temperature at 160°C for 10 minutes, then heating to 180°C for 5 minutes, and keeping the temperature at 180°C for 10 minutes; after the digestion is completed, driving out the acid at 180°C for 30 minutes, making up the remaining liquid to volume, shaking it evenly, letting it stand, filtering it, and using it as a test sample.

[0025] The content of each element can be determined by inductively coupled plasma tandem quadrupole mass spectrometry (ICP-MS / MS).

[0026] Preferably, the ICP-MS / MS operating parameters are: high-frequency plasma emission power 1550W; carrier gas is argon; concentric nebulizer: nebulizing gas flow rate 1.05L·min -1; Gas compensation gas flow 0.15L·min -1 ; Sampling depth 8.0mm; Peristaltic pump 0.3r·min -1 Ca, Na, Pb, and S elements can be detected using ICP-MS / MS-(KED mode), with helium as the collision gas and a flow rate of 4.5 mL min -1 .

[0027] The establishment of the identification model includes: establishing a stable isotope and element database of greenhouse-cultivated Tripterygium wilfordii and simulated wild-cultivated Tripterygium wilfordii samples, and screening out stable isotopes and elements with extremely significant differences between greenhouse-cultivated Tripterygium wilfordii and simulated wild-cultivated Tripterygium wilfordii with p<0.01 as variables through independent sample t-test comparison; using the principal component analysis method to screen the variables: extracting components with eigenvalues>1 in the principal component variance contribution to perform load analysis of each principal component variable, and selecting variables with absolute values ​​greater than 0.5 as preliminary variables; using partial least squares-discriminant analysis to screen out variables with important factors greater than 1 from the preliminary variables as stable isotopes and elements for constructing the identification model, and importing the stable isotopes and elements for constructing the identification model into a linear analysis function to obtain the identification model of Tripterygium wilfordii. Preferably, the operation of using partial least squares method-discriminant analysis to screen out variables with important factors greater than 1 from the preliminary selected variables can be performed more than twice, and the variables with important factors greater than 1 screened out in the last time are used as stable isotopes and elements for constructing the identification model; the accuracy of the identification model can be improved while reducing the detection workload.

[0028] The present invention has the following beneficial effects:

[0029] The present invention adopts specific stable isotope ratios and specific mineral element contents that are significantly different between simulated wild cultivated Tripterygium wilfordii and greenhouse cultivated Tripterygium wilfordii, especially specific stable isotope ratios and specific mineral element contents that are significantly different in different parts of Tripterygium wilfordii, and establishes a specific identification model suitable for leaves, stems, tuberous roots, and fibrous roots of Tripterygium wilfordii, which can accurately and efficiently identify simulated wild cultivated Tripterygium wilfordii and greenhouse cultivated Tripterygium wilfordii from leaves, stems, tuberous roots, and fibrous roots. The method can achieve an overall identification accuracy of 100% for simulated wild cultivated (wild tended) Tripterygium wilfordii and greenhouse cultivated (greenhouse tended) Tripterygium wilfordii.

[0030] The method of the present invention can be used with existing detection equipment without the need to use specific dedicated detection equipment, which is conducive to promotion and application. DETAILED DESCRIPTION

[0031] The present invention is further described in detail below in conjunction with specific embodiments so that those skilled in the art can better understand the technical solution of the present invention, but it should not be construed as limiting the technical solution of the present invention.

[0032] Example 1

[0033] 1. Pretreatment of the tested clover samples: process several samples of greenhouse-cultivated clover and simulated wild-cultivated clover separately, classify and dry the samples according to leaves, stems, tubers, and fibrous roots, crush them and pass them through a 100-mesh sieve, and collect the sieve-through as the clover samples.

[0034] Weigh 0.10g of the three-leaf green sample (100 mesh), place it in a sealed high-pressure digestion tank, add 6.0mL nitric acid, and after the sample is fully infiltrated by the acid solution, place it in 120℃ for pre-digestion for about half an hour until the yellow smoke dissipates. After cooling, cover the tank lid tightly and seal it, and install the digestion tank into the microwave digestion instrument. The digestion parameters are shown in Table 1. After the sample digestion is completed, take out the inner tank and drive the acid at 180℃ for half an hour, transfer about 0.5mL of the remaining liquid to a 25mL volumetric flask, make up the volume, shake well, let it stand, and filter it through a 0.45μm membrane to prepare the test sample for the determination of the content of each element.

[0035] Table 1 Microwave digestion program parameters of Tripterygium wilfordii plants

[0036]

[0037] 2. Analysis of multiple elements in Tripterygium wilfordii

[0038] 2.1 Instruments and Materials:

[0039] 8900 inductively coupled plasma tandem quadrupole mass spectrometer (ICP-MS / MS, Agilent, USA); TOPEX+ all-purpose microwave digestion instrument (Shanghai Yiyao Instrument Technology Development Co., Ltd.); BSA224S-CW electronic balance (Sartorius Scientific Instrument Co., Ltd.).

[0040] Single element standard solutions Ca (batch number GSB04-1720-2004), Na, Pb, S, etc. were purchased from the National Nonferrous Metals and Electronic Materials Analysis and Testing Center; nitric acid, hydrofluoric acid, and hydrochloric acid (super pure, CNW) were purchased from Shanghai Anpu Experimental Technology Co., Ltd., and perchloric acid (super pure, spaceship brand) was purchased from Tianjin Xinyuan Chemical Co., Ltd.; pure water was purchased from Guangzhou Watsons Group.

[0041] 2.2 Instrument conditions

[0042] ICP-MS / MS operating parameters: high-frequency plasma emission power 1550W; carrier gas is high-purity argon (99.999%); concentric nebulizer: nebulizing gas flow rate 1.05L·min -1 ; Gas compensation gas flow 0.15L·min -1 ; Sampling depth 8.0mm; Peristaltic pump 0.3r·min -1Ca, Na, Pb, and S were detected using ICP-MS / MS-(KED mode), with helium as the collision gas at a flow rate of 4.5 mL min -1 .

[0043] 3. Analysis of carbon, nitrogen, hydrogen and oxygen isotopes in Tripterygium wilfordii

[0044] 3.1 Instruments and Materials

[0045] Vario Pyro cube-Isoprime 100 elemental analysis-stable isotope ratio mass spectrometer (TC / EA-IRMS, Elementar, Germany) was used to determine δ 13 C and δ 15 Vario Isotope cube-Biovision elemental analysis-stable isotope ratio mass spectrometer (EA-IRMS, Elementar, Germany) was used to determine δD and δ 18 O; XPR106DUH / AC electronic balance (Mettler Toledo, Switzerland).

[0046] Granular reduced copper (analytical grade, Cu), tungsten trioxide (analytical grade, WO 3 ) and glassy carbon (analytical grade, C) were purchased from Elementar, Germany. The standard substances used included B2155 (δ 13 C=-26.98‰,δ 15 N=5.94‰)、USGS40(δ 15 N=-4.52‰)、IAEA-CH-6(δ 13 C=-10.449‰)、USGS64(δ 13 C=-40.82‰)、IAEA-N-2(δ 15 N=20.3‰), USGS54 (δD=-150.4‰, δ 18 O=17.79‰), USGS55 (δD=-28.2‰, δ 18 O=19.12‰), USGS56 (δD=-44.0‰, δ 18 O=27.23‰) were purchased from the International Atomic Energy Agency (IAEA) Joint Laboratory and subjected to multi-point calibration.

[0047] 3.2 Instrumental analysis

[0048] δ 13 C and δ 15N determination: Weigh 5.0 mg of Tripterygium wilfordii sample, wrap it in a tin foil cup and place it in the EA-IRMS sample tray. The carbon and nitrogen elements in the sample are converted into pure CO 2 and N 2 After the gas is discharged, it enters the gas isotope mass spectrometer for detection. Specific conditions for element analysis: The main filling material of the combustion tube is WO 3 , 1150℃; reduction tube filling is Cu, 850℃; high purity helium (He, 99.999%) as carrier gas. Main conditions for isotope mass spectrometry: CO 2 Sink current 100μA, N 2 The sink current is 400 μA. 2 The gas is diluted to achieve simultaneous measurement of two gases.

[0049] δD and δ 18 O determination: Weigh 0.5 mg of Tripterygium wilfordii sample, wrap it in a silver cup and place it in the instrument room for at least 48 hours for equilibrium, then place it in the TC / EA-IRMS sample tray. The hydrogen and oxygen elements in the sample are converted into pure H 2 After the reaction of the gas and CO, it enters the gas isotope mass spectrometer for detection. Specific conditions for elemental analysis: the main filler of the cracking tube is glassy carbon, the cracking temperature is 1450°C, and high-purity helium (He, 99.999%) is used as the carrier gas. The main conditions for isotope mass spectrometry: H 2 The trap current is 600 μA and the CO trap current is 200 μA. The CO gas is diluted using CentrION to achieve simultaneous measurement of the two gases.

[0050] The δ 13 C.δ 15 N, δD, δ 18 The formula for calculating the O stable isotope ratio is:

[0051] δE=(R 样品 / R 标准 -1);

[0052] Where, E is the target element isotope; R 样品 is the abundance ratio of heavy isotopes to light isotopes in the measured sample, that is, 13 C / 12 C. 15 N / 14 N. 3 H / 2 H. 18 O / 16 O, R 标准 is the abundance ratio of heavy isotopes to light isotopes in the standard sample; the δ value is multiplied by 1000 to obtain the final unit of ‰. The results are calibrated using international unified standard units, δ 13C uses the natural abundance relative to the VPDB standard, δ 15 N is calculated using the abundance, δD and δ 18 O uses natural abundance relative to the V-SMOW standard.

[0053] 4. Screening of identification model parameters for greenhouse cultivation and simulated wild cultivation of Tripterygium wilfordii

[0054] 4.1 Screening of stem discrimination parameters for greenhouse cultivation and simulated wild cultivation of Tripterygium wilfordii

[0055] Table 2 Isotope and element status of stems of Sanyeqing cultivated in greenhouse and simulated wild cultivation (mean ± SD)

[0056]

[0057]

[0058]

[0059] Note: After independent sample t-test comparison, p<0.05 represents that there are significant differences in the green stem elements of three leaves between greenhouse cultivation and simulated wild cultivation, and p<0.01 represents that there are extremely significant differences in the green stem elements of three leaves between greenhouse cultivation and simulated wild cultivation.

[0060] Through t-test statistics, 10 variables with extremely significant differences (p<0.01) were selected from the stable isotopes and elements tested, namely δ 13 C.δ 15 N, δD, Na, Ca, Ti, P, Ga, Tl, and Pb. The selected elements were used for further variable parameter screening using the principal component analysis method, as shown in Tables 3 and 4.

[0061] Table 3 Contribution of the variance of isotopes and elements in the stems of different planting methods of Tripterygium wilfordii

[0062]

[0063] Table 4 Loading analysis of the variables of stem isotopes and element principal components in different planting methods of Tripterygium wilfordii

[0064]

[0065]

[0066] According to Table 3, the principal component model established by isotopes and elements of stems of different planting methods of Tripterygium wilfordii extracted two principal components (eigenvalue>1), and the variance percentage of the first principal component far exceeded that of the second principal component. Therefore, the contribution of the variables in the first principal component was mainly considered. In Table 4, there are 8 variables with absolute values ​​greater than 0.5 in the first principal component, which is δ13 C.δ 15 N, Ca, Ti, P, Ga, Tl, Pb. In order to further screen the parameters of the discriminant model, the parameter screening method (important factor parameter) of the supervised model (partial least squares-discriminant analysis) was subsequently adopted. When the important factor was greater than 1, the parameter was selected as the construction variable of the final model. Table 5 shows the ranking of the important factor parameters of different cultivation methods. Two latent variables were screened from the supervised model. The important factors of each parameter in each latent variable were greater than 1. 13 C.δ 15 The three variables of N and Ca were used for the final model construction.

[0067] Table 5 Ranking of important factors of isotopes and elements of Tripterygium wilfordii in different planting methods

[0068] Latent variable 1 Latent variable 2 <![CDATA[δ 15 N]]> 1.2938 1.2864 Ca 1.1446 1.0568 <![CDATA[δ 13 C]]> 1.0952 1.0431 P 1.0557 0.9949 Pb 0.9998 0.9997 Ga 0.8475 0.9633 Ti 0.8016 0.8071 Tl 0.5712 0.7480

[0069] 4.2 Screening of parameters for distinguishing leaves of Tripterygium wilfordii from greenhouse cultivation and simulated wild cultivation

[0070] Table 6 Isotope and element status of leaves of Tripterygium wilfordii grown in greenhouse and simulated wild (mean ± SD)

[0071]

[0072]

[0073] Note: After independent sample t-test comparison, p<0.05 represents that there are significant differences in the leaf elements of the three-leaf green grass cultivated in the greenhouse and the simulated wild cultivation, and p<0.01 represents that there are extremely significant differences in the leaf elements of the three-leaf green grass cultivated in the greenhouse and the simulated wild cultivation.

[0074] Through t-test statistics, 12 variables with extremely significant differences (p<0.01) were selected from the stable isotopes and elements tested, namely δ 13 C.δ 15 N, Ca, S, Rb, Se, Cd, Pr, Nd, Sm, Pb, and Bi. The selected isotopes and elements were used for further variable parameter screening using the principal component analysis method, as shown in Tables 7 and 8.

[0075] Table 7 Variance contribution of isotopes and elements in leaves of Tripterygium wilfordii under different planting methods

[0076]

[0077] Table 8 Analysis of the variables loading of leaf isotopes and element principal components in different planting methods of Tripterygium wilfordii

[0078] variable Ingredients 1 Ingredient 2 Sm 0.941 0.275 Pb 0.916 -0.067 Pr 0.912 0.347 Nd 0.910 0.357 Se 0.905 0.338 Cd 0.853 -0.019 Bi 0.757 -0.338 <![CDATA[δ 15 N]]> -0.754 -0.087 R 0.753 -0.438 Ca -0.743 0.160 <![CDATA[δ 13 C]]> -0.700 0.585 S -0.637 0.126

[0079] According to Table 7, the principal component model established by isotopes and elements of leaves of different planting methods of Tripterygium wilfordii extracted two principal components (eigenvalue>1), and the variance percentage of the first principal component far exceeded that of the second principal component. Therefore, the contribution of the variables in the first principal component was mainly considered. In Table 8, there are still 12 variables with absolute values ​​greater than 0.5, which is δ 13 C.δ 15 N, Ca, S, Rb, Se, Cd, Pr, Nd, Sm, Pb, Bi. In order to further screen the parameters of the discriminant model, the parameter screening method (important factor parameter) of the supervised model (partial least squares-discriminant analysis) was subsequently used. When the important factor was greater than 1, the parameter was selected as the construction variable of the final model. Table 9 shows the ranking of the important factor parameters of different cultivation methods. Two latent variables were screened from the supervised model. The important factors of each parameter in each latent variable were greater than 1, namely Pb, δ 15 The three variables of N and S are used for the final model construction.

[0080] Table 9 Ranking of important factors of isotopes and elements in leaves of Tripterygium wilfordii under different planting methods

[0081] Latent variable 1 Latent variable 2 Pb 1.2841 1.1893 <![CDATA[δ 15 N]]> 1.2708 1.3690 S 1.0621 1.1073 R 1.0546 0.9602 Cd 1.0521 0.9236 Sm 1.0160 0.9575 Ca 0.9445 0.8301 Nd 0.9015 0.9554 Pr 0.8787 0.9779 Se 0.8683 0.9636 <![CDATA[δ 13 C]]> 0.8286 0.7389 Bi 0.6629 0.8722

[0082] 4.3 Screening of root discrimination parameters between greenhouse cultivation and simulated wild cultivation of Tripterygium wilfordii

[0083] Table 10 Isotope and element status of root tubers of Tripterygium wilfordii grown in greenhouse and simulated wild cultivation (mean ± SD)

[0084]

[0085]

[0086] Note: After independent sample t-test comparison, p<0.05 represents that there are significant differences in the root elements of the greenhouse-cultivated and simulated wild-cultivated Tripterygium wilfordii, and p<0.01 represents that there are extremely significant differences in the root elements of the greenhouse-cultivated and simulated wild-cultivated Tripterygium wilfordii.

[0087] Through t-test statistics, 7 variables with extremely significant differences (p<0.01) were selected from the stable isotopes and elements tested, namely δ 13 C.δ 15 N, Na, Ca, S, Rb, Bi, the screened elements were used for further variable parameter screening using the principal component analysis method, as shown in Tables 11 and 12.

[0088] Table 11 Contribution of variance of isotopes and elements in the tubers of Tripterygium wilfordii under different planting methods

[0089]

[0090] Table 12 Analysis of the variables loading of isotopes and element principal components of root tubers in different planting methods of Tripterygium wilfordii

[0091] parameter Ingredients 1 Na 0.913 S 0.883 R -0.553 <![CDATA[δ 13 C]]> 0.823 Ca 0.749 Bi -0.486 <![CDATA[δ 15 N]]> 0.661

[0092] According to Table 11, the principal component model established by isotopes and elements of tubers of different planting methods extracted one principal component (eigenvalue>1). In Table 12, there are 5 variables with absolute values ​​greater than 0.5 in the first principal component (the absolute value of Rb is slightly greater than 0.5 and is not included), which is δ 13 C.δ 15 N, Ca, S, Na. In order to further screen the parameters of the discriminant model, the parameter screening method (important factor parameter) of the supervised model (partial least squares-discriminant analysis) was subsequently used. When the important factor was greater than 1, the parameter was selected as the construction variable of the final model. Table 13 shows the ranking of the important factor parameters of different cultivation methods. One latent variable was screened from the supervised model. The important factors of each parameter greater than 1 were Na, δ 13 C.δ 15 The three variables of N are used for the final model construction.

[0093] Table 13 Ranking of important factors of isotopes and elements in root tubers of Tripterygium wilfordii under different planting methods

[0094] parameter Latent variable 1 Na 1.2938 <![CDATA[δ 13 C]]> 1.1446 <![CDATA[δ 15 N]]> 1.0952 S 0.9557 Ca 0.9998

[0095] 4.4 Screening of fibrous root discrimination parameters between greenhouse cultivation and simulated wild cultivation of Tripterygium wilfordii

[0096] Table 14 Isotope and element status of fibrous roots of Tripterygium wilfordii grown in greenhouse and simulated wild cultivation (mean ± SD)

[0097]

[0098]

[0099] Note: After independent sample t-test comparison, p<0.05 represents that there are significant differences in the fibrous root elements of the greenhouse-cultivated and simulated wild-cultivated Tripterygium wilfordii, and p<0.01 represents that there are extremely significant differences in the fibrous root elements of the greenhouse-cultivated and simulated wild-cultivated Tripterygium wilfordii.

[0100] Through t-test statistics, 12 variables with extremely significant differences (p<0.01) were selected from the stable isotopes and elements tested, namely δ 13 C.δ 15 N, Mg, Ca, Zn, Ga, Sr, Mo, Ce, Pr, Nd, and Ho. The screened elements were used for further variable parameter screening using the principal component analysis method, as shown in Tables 15 and 16.

[0101] Table 15 Variance contribution of isotopes and elements in the fibrous roots of different planting methods of Tripterygium wilfordii

[0102]

[0103] Table 16 Analysis of the variable loadings of isotopes and elemental principal components of fibrous roots in different planting methods of Tripterygium wilfordii

[0104]

[0105]

[0106] According to Table 15, the principal component model established by isotopes and elements of fibrous roots of different planting methods extracted three principal components (eigenvalue>1), and the variance percentage of the first principal component far exceeded that of the second and third principal components. Therefore, the contribution of the variables in the first principal component was mainly considered. In Table 16, there are 9 variables with absolute values ​​greater than 0.5, which are δ 13 C.δ 15 N, Ca, Ga, Nd, Ce, Mg, Ho, Zn. In order to further screen the parameters of the discriminant model, the parameter screening method (important factor parameter) of the supervised model (partial least squares-discriminant analysis) was used. When the important factor was greater than 1, the parameter was selected as the construction parameter of the final model. Table 17 shows the ranking of the important factor parameters of different cultivation methods. The supervised model was screened twice and finally selected Ca, δ 13 C.δ 15 The three parameters of N are used for the final model construction.

[0107] Table 17 Ranking of important factors of isotopes and elements in the fibrous roots of Tripterygium wilfordii under different planting methods

[0108]

[0109] 5. Identification model of greenhouse cultivation and simulated wild cultivation of Tripterygium wilfordii

[0110] In order to explore the differences between greenhouse-reared and wild-reared Tripterygium wilfordii, the project collected 40 samples of wild and greenhouse-reared Tripterygium wilfordii from different origins, and used stable isotope and mineral element technology to study their inherent correlation. This experiment determined the isotope values ​​and element contents in the leaves, stems, tubers and fibrous roots of Tripterygium wilfordii. Through parameter screening, the final model parameters were determined respectively. The linear discriminant analysis method was used to analyze wild and greenhouse Tripterygium wilfordii, and an identification model for different growth modes of Tripterygium wilfordii was established. The basic idea of ​​this method is to find a suitable projection axis so that the overlapping parts of the projections of samples of different categories on the axis are minimized, thereby achieving the best classification effect. The general principle of the identification model is:

[0111] A discriminant function is constructed with the help of variance analysis, namely:

[0112] F = k 1 X 1 +k 2 X 2 +……+k p X p ; coefficient k 1 , k 2 ……k p The principle of determination is to maximize the gap between groups and minimize the gap within groups. The judgment criterion is: suppose the population is divided into p categories, if

[0113] |F p -F 1 |<|F p -F q |(p, q=1, 2, ... m, q=1), then F p Belongs to the first category.

[0114] 5.1 Leaf Discriminant Model

[0115] The δ of leaves from the two cultivation methods 15 The elements N, Pb (mg / kg), and S (mg / kg) were introduced into the linear analysis function to obtain the identification model of the leaves of Tripterygium wilfordii:

[0116] F 大棚叶 =2.462δ 15 N+2.990Pb+0.022S-36.876

[0117] F 仿野生叶 =0.584δ 15 N+14.090Pb+0.016S-25.508

[0118] 5.2 Stem discrimination model

[0119] The δ of the stems of the two cultivation methods 13 C.δ 15 The N value and Ca (mg / kg) element were introduced into the linear analysis function to obtain the identification model of the stem of Tripterygium wilfordii:

[0120] F 大棚茎 =0.652δ 15 N+0.007Ca-21.506δ 13 C-359.658

[0121] F 仿野生茎 =-0.898δ 15 N+0.006Ca-22.990δ 13 C-390.142

[0122] 5.3 Root tuber identification model

[0123] The δ of the tuberous roots of the two cultivation methods was 13 C.δ 15 The N value and the Na content (mg / kg) were imported into the linear analysis function to obtain the identification model of the tuberous roots of Tripterygium wilfordii:

[0124] F 大棚块根 =4.599δ 15 N+0.051Na-19.317δ 13 C-297.410

[0125] F 仿野生块根 =3.942δ 15 N+0.044Na-19.926δ 13 C-311.255

[0126] 5.4 Root Discrimination Model

[0127] The δ of fibrous roots of the two cultivation methods was 13 C.δ 15 The N value and Ca content (mg / kg) were imported into the linear analysis function to obtain the identification model of the fibrous roots of Tripterygium wilfordii:

[0128] F 大棚须根 =-9.164δ 15 N+0.010Ca-38.071δ 13 C-600.163

[0129] F 仿野生须根 =-11.746δ 15 N+0.010Ca-41.321δ 13 C-694.687

[0130] 5.5 Model Validation

[0131] The model was back-tested and verified, that is, the corresponding parameter content of each sample was substituted into the model of greenhouse cultivated Tripterygium wilfordii and simulated wild cultivated Tripterygium wilfordii for calculation. 大棚茎 >F 仿野生茎 , F 大棚叶 >F 仿野生叶 , F 大棚块根 >F 仿野生块根 or F 大棚须根 >F 仿野生须根 , then the sample is judged to be greenhouse-cultivated Tripterygium wilfordii, otherwise it is simulated wild-cultivated Tripterygium wilfordii. This is the discrimination criterion given when establishing the identification model. Simply put, the larger this value is, the closer the sample point is to this category, and the greater the probability of belonging to this category.

[0132] Example 2

[0133] Two root samples of Tripterygium wilfordii were taken, and the stable carbon and nitrogen isotope ratios and the mineral element Ca content are shown in Table 18.

[0134] Table 18 Stable carbon and nitrogen isotope ratios and mineral element Ca content in the root samples of Tripterygium wilfordii

[0135]

[0136] Substitute the stable carbon and nitrogen isotope ratios and Ca content of sample 1 into the two identification models of Tripterygium wilfordii root to obtain F 大棚须根 =601.82 and F 仿野生须根 =587.41, that is, F 大棚须根 >F 仿野生须根 Therefore, it was determined that Sample 1 was the fibrous roots of Tripterygium wilfordii cultivated in a greenhouse, which was consistent with Sample 1 actually obtained from Tripterygium wilfordii cultivated in a greenhouse.

[0137] Substitute the stable carbon and nitrogen isotope ratios and Ca content of sample 2 into the two identification models of Tripterygium wilfordii root, and obtain F 大棚须根 =790.68 and F 仿野生须根 =807.87, that is, F 大棚须根 <F 仿野生须根 Therefore, it was determined that Sample 2 was the fibrous root of simulated wild-cultivated Tripterygium wilfordii, which was consistent with Sample 2 actually obtained from simulated wild-cultivated Tripterygium wilfordii.

[0138] It can be seen that the accuracy of the method of the present invention for identifying the simulated wild-cultivated Tripterygium wilfordii and the greenhouse-cultivated Tripterygium wilfordii can reach 100%.

[0139] Example 3

[0140] Two leaf samples of Tripterygium wilfordii were taken, and the stable nitrogen isotope ratio, Pb element content and S element content are shown in Table 19.

[0141] Table 19 Stable nitrogen isotope ratios, mineral element Pb and S content in the samples of Tripterygium wilfordii leaves

[0142]

[0143] Substitute the stable nitrogen isotope ratio, Pb content and S content of sample 3 into the two identification models of the leaves of Tripterygium wilfordii to obtain F 大棚叶 =23.36 and F 仿野生叶 =14.72, that is, F 大棚叶 >F 仿野生叶 Therefore, it was determined that sample 3 was the leaf of Tripterygium wilfordii cultivated in a greenhouse, which was consistent with sample 3 actually obtained from Tripterygium wilfordii cultivated in a greenhouse.

[0144] Substitute the stable nitrogen isotope ratio, Pb content and S content of sample 4 into the two identification models of the leaves of Tripterygium wilfordii to obtain F 大棚叶 =12.41 and F 仿野生叶 =25.60, that is, F 大棚叶 <F 仿野生叶 Therefore, sample 4 was judged to be the leaf of simulated wild-cultivated Tripterygium wilfordii, which was consistent with sample 4 actually obtained from simulated wild-cultivated Tripterygium wilfordii.

[0145] It can be seen that the accuracy of the method of the present invention for identifying the simulated wild-cultivated Tripterygium wilfordii and the greenhouse-cultivated Tripterygium wilfordii can reach 100%.

[0146] Example 4

[0147] Two root tuber samples of Tripterygium wilfordii were taken, and the stable carbon and nitrogen isotope ratios and the mineral element Na content are shown in Table 20.

[0148] Table 20 Stable carbon and nitrogen isotope ratios and mineral element Na content in root samples of Tripterygium wilfordii

[0149]

[0150] Substitute the stable carbon and nitrogen isotope ratios and the Na content of sample 5 into the two identification models of the tuberous roots of Tripterygium wilfordii to obtain F 大棚块根 =284.40 and F 仿野生块根 =280.56, that is, F 大棚块根 >F 仿野生块根 Therefore, it was determined that sample 5 was the tuberous root of Tripterygium wilfordii cultivated in a greenhouse, which was consistent with sample 5 actually obtained from Tripterygium wilfordii cultivated in a greenhouse.

[0151] Substituting the stable carbon and nitrogen isotope ratios and the Na content of sample 6 into the two identification models of the tuberous roots of Tripterygium wilfordii, we obtain F 大棚块根 =294.30 and F 仿野生块根 =296.67, that is, F 大棚块根 <F 仿野生块根 Therefore, it was determined that sample 6 was the tuberous root of simulated wild-cultivated Tripterygium wilfordii, which was consistent with sample 6 actually obtained from simulated wild-cultivated Tripterygium wilfordii.

[0152] It can be seen that the accuracy of the method of the present invention for identifying the simulated wild-cultivated Tripterygium wilfordii and the greenhouse-cultivated Tripterygium wilfordii can reach 100%.

[0153] Example 5

[0154] Two stem samples of Tripterygium wilfordii were taken, and the stable carbon and nitrogen isotope ratios and the mineral element Ca content are shown in Table 21.

[0155] Table 21 Stable carbon and nitrogen isotope ratios and mineral element Ca content in the stem samples of Tripterygium wilfordii

[0156]

[0157] Substituting the stable carbon and nitrogen isotope ratios and Ca content of sample 7 into the two identification models of the stem of Tripterygium wilfordii, we obtained F 大棚茎 =343.72 and F 仿野生茎 =337.55, that is, F 大棚茎 >F 仿野生茎 Therefore, sample 7 was judged to be the stem of Tripterygium wilfordii cultivated in a greenhouse, which was consistent with sample 7 actually obtained from Tripterygium wilfordii cultivated in a greenhouse.

[0158] Substituting the stable carbon and nitrogen isotope ratios and Ca content of sample 8 into the two identification models of the stem of Tripterygium wilfordii, we obtain F 大棚茎 =331.53 and F 仿野生茎 =341.03, that is, F 大棚茎 <F 仿野生茎 Therefore, it was determined that sample 8 was the stem of simulated wild-cultivated Tripterygium wilfordii, which was consistent with sample 8 actually obtained from simulated wild-cultivated Tripterygium wilfordii.

[0159] It can be seen that the accuracy of the method of the present invention for identifying the simulated wild-cultivated Tripterygium wilfordii and the greenhouse-cultivated Tripterygium wilfordii can reach 100%.

[0160] Comparative Example 1

[0161] The δ of the fibrous roots of the two cultivation methods in Example 1 13 The C value and Ca content (mg / kg) were introduced into the linear analysis function to obtain the comparative identification model of the fibrous roots of Tripterygium wilfordii. The two fibrous root samples of Tripterygium wilfordii in Example 2 were taken, and the stable carbon isotope ratio and Ca content of sample 1 were respectively substituted into the two comparative identification models of the fibrous roots of Tripterygium wilfordii to obtain F. 大棚须根 <F 仿野生须根 Therefore, it is determined that sample 1 is the simulated wild cultivated Tripterygium wilfordii root, which is inconsistent with sample 1 obtained from the Tripterygium wilfordii cultivated in the greenhouse. The stable carbon isotope ratio and Ca content of sample 2 are respectively substituted into the two comparative identification models of Tripterygium wilfordii root, and F is obtained. 大棚须根 <F 仿野生须根 , so it is determined that sample 2 is the simulated wild cultivated Tripterygium wilfordii root, which is consistent with the sample 2 actually obtained from the simulated wild cultivated Tripterygium wilfordii. 13 The comparative identification model established based on the C and Ca element content (mg / kg) has an accuracy rate of only 50% for identifying the simulated wild-cultivated Tripterygium wilfordii and the greenhouse-cultivated Tripterygium wilfordii, with a large error.

[0162] The identification model of greenhouse-cultivated clover and simulated wild-cultivated clover established by the present invention can be used to identify greenhouse-cultivated clover and simulated wild-cultivated clover, and the authenticity of simulated wild-cultivated clover products can be identified. The method of the present invention has good repeatability, fast analysis speed, simple operation, and the authenticity identification rate of simulated wild-cultivated clover can reach 100%, which is a fast, economical and practical identification technology.

Claims

1. A method for identifying wild-cultivated Tripterygium wilfordii and greenhouse-cultivated Tripterygium wilfordii, characterized in that: It includes using one or more of the leaves, stems, tubers and fibrous roots of the tested clover samples to identify the simulated wild cultivated clover and the greenhouse cultivated clover; The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the leaves of the tested Tripterygium wilfordii sample comprises: determining the δ 15 The N stable isotope ratio, Pb element content and S element content were respectively substituted into the following identification model of the leaves of the three-leaf green for calculation, and the F 大棚叶 and F 仿野生叶 If F 大棚叶 >F 仿野生叶 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生叶 >F 大棚叶 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii; F 大棚叶 =2.462δ 15 N+2.990Pb+0.022S-36.876 F 仿野生叶 =0.584δ 15 N+14.090Pb+0.016S-25.508; The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the stem of the tested Tripterygium wilfordii sample comprises: determining the δ 13 C stable isotope ratio, δ 15 The N stable isotope ratio and Ca content were substituted into the following identification model of the stem of Tripterygium wilfordii for calculation, and the F 大棚茎 and F 仿野生茎 If F 大棚茎 >F 仿野生茎 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生茎 >F 大棚茎 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii; F 大棚茎 =0.652d 15 N+0.007Ca-21.506d 13 C-359,658 F 仿野生茎 =-0.898δ 15 N+0.006Ca-22.990δ 13 C-390.142; The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the tubers of the tested Tripterygium wilfordii sample comprises: determining the δ 13 C stable isotope ratio, δ 15 The N stable isotope ratio and Na content were substituted into the following identification model of the root of Tripterygium wilfordii for calculation, and the F 大棚块根 and F 仿野生块根 If F 大棚块根 >F 仿野生块根 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生块根 >F 大棚块根 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii; F 大棚块根 =4.599δ 15 N+0.051Na-19.317δ 13 C-297.410 F 仿野生块根 =3.942δ 15 N+0.044Na-19.926δ 13 C-311.255; The method for identifying the simulated wild cultivated Tripterygium wilfordii and the greenhouse cultivated Tripterygium wilfordii by using the fibrous roots of the tested Tripterygium wilfordii sample comprises: determining the δ 13 C stable isotope ratio, δ 15 The N stable isotope ratio and Ca element content were substituted into the following identification model of the root hairs of Tripterygium wilfordii for calculation, and the F 大棚须根 and F 仿野生须根 If F 大棚须根 >F 仿野生须根 , then the sample is judged to be greenhouse-grown Tripterygium wilfordii. On the contrary, if F 仿野生须根 >F 大棚须根 , then the sample is judged to be simulated wild cultivated Tripterygium wilfordii; F 大棚须根 =-9.164δ 15 N+0.010Ca-38.071δ 13 C-600.163 F 仿野生须根 =-11.746δ 15 N+0.010Ca-41.321δ 13 C-694.687; The content of each element is expressed in mg / kg, and the stable isotope ratio is expressed in ‰.

2. The identification method according to claim 1, characterized in that: The stable isotope ratio calculation formula is: δE=(R 样品 / R 标准 -1); Where, E is the target element isotope; R 样品 is the abundance ratio of heavy isotopes to light isotopes in the measured sample, that is, 13 C / 12 C. 15 N / 14 N, R 标准 is the abundance ratio of heavy isotopes to light isotopes in the standard sample; the δ value is multiplied by 1000 to obtain the final unit ‰, and the result is calibrated using the international unified standard unit, δ 13 C uses the natural abundance relative to the VPDB standard, δ 15 N refers to the abundance of nitrogen relative to air.

3. The identification method according to claim 2, characterized in that: The isotope abundance of each sample was determined by elemental analysis-stable isotope ratio mass spectrometry.

4. The identification method according to claim 3, characterized in that: δ 13 C.δ 15 The conditions for elemental analysis during N determination were as follows: the combustion tube was mainly filled with WO3, 1150°C; the reduction tube was filled with Cu, 850°C; helium was used as the carrier gas; the conditions for stable isotope ratio mass spectrometry were as follows: CO2 trap current 100μA, N2 trap current 400μA.

5. The identification method according to claim 1, characterized in that: The sample of the three-leaf green to be tested is first pretreated and then the content of each element is determined. The pretreatment includes: classifying the sample of the three-leaf green to be tested according to leaves, stems, tubers, and fibrous roots and drying them, crushing them and passing them through a 100-mesh sieve, collecting the sieve under the sieve and placing them in a high-pressure digestion tank, adding nitric acid, and after the sample is fully infiltrated by the acid solution, placing it in a 120°C pre-digestion until the yellow smoke dissipates, cooling it and sealing it, placing it in a microwave digestion instrument for digestion, and the digestion conditions are: first heating to 120°C for 5 minutes, keeping the temperature at 120°C for 5 minutes, then heating to 160°C for 5 minutes, keeping the temperature at 160°C for 10 minutes, then heating to 180°C for 5 minutes, and keeping the temperature at 180°C for 10 minutes; after the digestion is completed, driving out the acid at 180°C for 30 minutes, making up the remaining liquid to volume, shaking it evenly, letting it stand, filtering it, and using it as a test sample.

6. The identification method according to claim 1 or 5, characterized in that: The contents of each element were determined by inductively coupled plasma quadrupole mass spectrometry.

7. The identification method according to claim 1, characterized in that: The establishment of the identification model includes: establishing a stable isotope and element database of greenhouse-cultivated Tripterygium wilfordii and simulated wild-cultivated Tripterygium wilfordii samples, and screening out stable isotopes and elements with extremely significant differences between greenhouse-cultivated Tripterygium wilfordii and simulated wild-cultivated Tripterygium wilfordii with p<0.01 as variables through independent sample t-test comparison; using the principal component analysis method to screen the variables: extracting components with eigenvalues>1 in the principal component variance contribution to perform load analysis of each principal component variable, and selecting variables with absolute values ​​greater than 0.5 as preliminary variables; using partial least squares-discriminant analysis to screen out variables with important factors greater than 1 from the preliminary variables as stable isotopes and elements for constructing the identification model, and importing the stable isotopes and elements for constructing the identification model into a linear analysis function to obtain the identification model of Tripterygium wilfordii.

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