Method for detecting authenticity and content of torreya grandis seed oil based on characteristics of 1, 1.4 a-trimethyl-7-isopropyl-1, 2, 3. 4 a, 9.10. 10 a-octahydrophenanthrene

Abstract

The invention relates to the technical field of quality control of torreya grandis seed oil, and discloses a method for detecting authenticity and content of torreya grandis seed oil based on characteristics of 1, 1, 4a-trimethyl-7-isopropyl-1, 2, 3, 4, 4a, 9, 10, 10a-octahydrophenanthrene, and the authenticity and content of the torreya grandis seed oil are detected by analyzing the existence and content of the 1, 1, 4a-trimethyl-7-isopropyl-1, 2, 3, 4, 4a, 9, 10, 10a-octahydrophenanthrene in vegetable oil. Performing qualitative judgment and quantitative analysis on the torreya grandis seed oil; the method comprises the following steps: extracting, purifying and measuring 1, 1, 4a-trimethyl-7-isopropyl-1, 2, 3, 4, 4a, 9, 10, 10a-octahydrophenanthrene in the vegetable oil by adopting the technologies of dissolving the vegetable oil with n-hexane, degreasing with a sulfonation method, deacidifying and purifying with a sodium sulfate aqueous solution, detecting with a gas chromatograph-mass spectrometer and the like, and determining whether the 1, 1, 4a-trimethyl-7-isopropyl-1, 2, 3, 4, 4a, 9, 10, 10a-octahydrophenanthrene exists in the vegetable oil or not and the content of the 1, 1, 4a-trimethyl-7-isopropyl-1, 2, 3, 4, 4a, 9, 10, 10a-octahydrophenanthrene in the vegetable oil. The authenticity and the content of the torreya grandis seed oil are indicated. The method is simple and feasible in technology and good in reproducibility, can realize accurate identification and content analysis of the torreya grandis seed oil, and has a good application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the quality control technology field of torreya grandis seed oil, and in particular to a method for detecting the authenticity and content of torreya grandis seed oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene characteristics. BACKGROUND

[0002] Torreya grandis, also known as yew nut, is a plant of Taxaceae, mainly produced in Zhejiang Province. There are wild Torreya trees in 25 counties (cities) of the province, with a total resource of over 600,000 trees, including more than 480,000 mature trees and more than 50,000 ancient trees of over 100 years old. As a characteristic economic tree species in Zhejiang, Torreya grandis is the core income source for Torreya farmers and the pillar industry of regional agriculture and forestry economy. Under the policy of "Torreya grandis southward expansion" and "Torreya grandis westward expansion", the cultivation area has increased to 800,000 mu, and the yield has jumped from 1,000 tons in 2006 to more than 8,000 tons in 2020. However, the industry is highly dependent on traditional roasted Torreya seed products, and the single processing method leads to oversupply, with the price falling from a peak of 300 yuan / kg to 50 yuan / kg (2020). The problem of "increased production but not increased income" is highlighted. Most of the newly added Torreya trees are young trees, and the future production will continue to rise, exacerbating the supply and demand imbalance. Therefore, the development of high-value and healthy products (such as Torreya seed oil) has become a key path for the sustainable development of the industry.

[0003] Torreya grandis kernel contains oil at a rate of 42% to 61%, with unsaturated fatty acids accounting for more than 80%. Abietic acid (5,11,14-eicosatrienoic acid) as a characteristic functional ingredient, the content is 4.3% to 12.3%. Animal experiments show that Torreya seed oil can significantly reduce the total cholesterol (TC), triglyceride (TG) and atherosclerosis index (AI) of Wistar rats, and has clear health benefits. Due to its excellent quality, the market price of Torreya seed oil has been higher than 600 yuan / kg for a long time. However, the current "Edible Vegetable Oil Hygiene Standard" (GB 2716-2018) cannot effectively control the quality of Torreya seed oil, leading to an increasing risk of adulteration driven by high profits.

[0004] Current methods for identifying the authenticity of vegetable oil mainly include: chromatography (gas / liquid chromatography, such as GC-MS for determining fatty acids); spectroscopy (infrared, fluorescence, ultraviolet-visible spectroscopy, etc.); combined techniques (chromatography-mass spectrometry, carbon isotope ratio mass spectrometry); rapid detection methods (color reaction, electronic nose / tongue). Most of the above methods rely on functional groups or characteristic substances (such as sterols, volatile components) in crude oil, but Torreya seed oil is mainly refined oil, and the characteristic components may be removed or their content fluctuates during processing, causing the traditional methods to fail. Compared with the specific identification research of pecan seed oil, oil tea seed oil and Torreya seed oil, there is a lack of research.

[0005] Therefore, the inventors focus on the stable and refined-tolerant octahydrophenanthrene derivative marker in torreya grandis seed oil, which can remain after processing and provide reliable basis for authenticity discrimination and quantitative blending, and fill the blank of the prior art. SUMMARY

[0006] In view of the deficiencies in the prior art, the purpose of the present application is to provide a method for determining the authenticity and content of torreya grandis seed oil using 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene as an index, to solve the technical problem of lack of analysis of torreya grandis seed oil content in the finished product "torreya grandis seed oil" in the prior art.

[0007] In order to achieve the above-mentioned purpose, the present application provides the following technical scheme: A method for detecting the authenticity and content of torreya grandis seed oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, comprising the following steps: A. Mix the plant oil sample uniformly, and weigh the plant oil sample into a test tube with a plug; B. Add n-hexane to dissolve the plant oil in the test tube containing the plant oil sample, cover the plug and mix uniformly at high speed; C. Add concentrated sulfuric acid to the mixed test tube, cover the plug and mix uniformly at high speed; D. Place the vortexed test tube on a test tube rack for static treatment and layering to remove fatty acids, pigments and impurities in the plant oil; E. Absorb the supernatant (n-hexane layer) after static treatment into another test tube with a plug, add sodium sulfate aqueous solution, cover the plug and vortex at high speed; F. Place the vortexed test tube on a test tube rack for static treatment and layering to remove residual sulfuric acid in the n-hexane extract; G. Absorb 1ml of the supernatant (n-hexane layer) after static treatment in F, pass through a microporous filter membrane and then into a sample bottle, and then detect and analyze it on a gas chromatograph-mass spectrometer; H. Establish a calibration curve for the percentage content of torreya grandis seed oil: weigh the self-made and pressed torreya grandis seed oil, supplement the plant oil to 1g if it is less than 1g, prepare torreya grandis seed oil with mass fractions of 0, 10%, 30%, 50%, 70% and 100%, respectively, and analyze them simultaneously according to steps B to G, and establish a calibration curve for the mass percentage concentration of torreya grandis seed oil with the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene as the ordinate and the mass percentage of torreya grandis seed oil as the abscissa; I. The sample is verified whether the vegetable oil contains torreya grandis seed oil by detecting 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, and the mass percentage concentration of the torreya grandis seed oil in the sample is calculated by using the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene and the calibration curve established in step H.

[0008] The present application obtains an octahydrophenanthrene derivative, i.e., 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene (C 20 H 30 , CAS No. 1000210-28-9), by screening characteristic components of torreya grandis seed oil products from different sources, oil extraction methods and processing degrees, and the mass spectrum thereof is shown in Figure 1 The component only exists in torreya grandis seed oil among common edible vegetable oils, and the content deviation of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is only 5% by determining 20 torreya grandis seed oils with different processing methods, which shows that the content of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is relatively stable in crude oil and refined oil of torreya grandis seed oil. The present application qualitatively determines and quantitatively analyzes torreya grandis seed oil by analyzing the presence or absence and content of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in vegetable oil.

[0009] Meanwhile, vegetable oil is a kind of oil and fat mainly composed of fatty acid glycerides, rich in vitamins, pigments, proteins, terpenes and other substances, and the components are extremely complex. If the pretreatment is not ideal, it will directly affect the detection results of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene by subsequent instruments. The degreasing treatment is extremely critical, and the present method adopts sulfonation method for degreasing and impurity removal, and sodium sulfate aqueous solution for deacidification and purification, so as to achieve ideal purification effect.

[0010] As preferred, 0.5-1.5 g of the vegetable oil sample is weighed in a 25 ml test tube with a plug in step A, and preferably the mass of the vegetable oil sample is 1.00 g.

[0011] As preferred, chromatographically pure n-hexane is used as n-hexane in step B, 10 ml of n-hexane is added, and the vortex time is 10-30 s, preferably 30 s.

[0012] As preferred, concentrated sulfuric acid of superior grade is used in step C, the volume is 3-5 ml, preferably 3 ml, and the plug is covered and vortexed at high speed for 20 s.

[0013] As preferred, the standing layer separation time in steps D and F is 10-20 min, preferably 15 min.

[0014] As preferred, the volume of the supernatant sucked in step E is 3-5 ml, preferably 5 ml; the added sodium sulfate aqueous solution is 10 ml of 2% mass concentration of anhydrous sodium sulfate aqueous solution, and high-speed vortex is performed for 20 s.

[0015] As preferred, the volume of the supernatant sucked in step G is 1-2 ml, preferably 1 ml; the pore size of the microporous filter membrane is 0.25 μm.

[0016] As preferred, the plant oil supplemented in step H is soybean oil.

[0017] The present application has the following technical effects due to the above technical solutions: The present application is based on the fact that 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene only exists in torreya grandis seed oil among common edible plant oils, and the content of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in torreya grandis seed oil of different sources and processing stages is relatively stable. The presence or absence and content of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in plant oil are analyzed to qualitatively determine and quantitatively analyze torreya grandis seed oil. Meanwhile, the sulfonation method is used to remove fat and impurities, and the sodium sulfate aqueous solution is used to remove acid to purify the gas chromatograph-mass spectrometer for detecting 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in plant oil. The presence or absence and content of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in plant oil indicate the authenticity and content of torreya grandis seed oil. The method is simple and feasible, has good reproducibility, can realize accurate identification and content analysis of torreya grandis seed oil, and has good application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The mass spectrum of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene.

[0019] Figure 2 The total ion current chromatogram of the sample in Example 1 (containing the part of “1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene”).

[0020] Figure 3Mass Spectrum of 1,1,4a-Trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene in Example 1 Sample (where the top half is the sample spectrum and the bottom half is the spectrum of 1,1,4a-Trimethyl-7-isopropyl- 1,2,3,4,4a,9,10,10a-octahydrophenanthrene from the NIST 20 library).

[0021] Figure 4 Selected Ion Spectrum of 1,1,4a-Trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene in Example 1 Sample.

[0022] Figure 5 Total Ion Chromatogram of Example 2 Sample (with "1,1,4a-Trimethyl-7- isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene" should be present).

[0023] Figure 6 Mass Spectrum of 1,1,4a-Trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene in Example 2 Sample (where the top half is the sample spectrum and the bottom half is the spectrum of 1,1,4a-Trimethyl-7-isopropyl- 1,2,3,4,4a,9,10,10a-octahydrophenanthrene from the NIST 20 library).

[0024] Figure 7 Selected Ion Spectrum of 1,1,4a-Trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene in Example 2 Sample.

[0025] Figure 8 Total Ion Chromatogram of Example 3 Sample (with "1,1,4a-Trimethyl-7- isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene" should be present).

[0026] Figure 9 Selected Ion Mass Spectrum of 1,1,4a-Trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene in Example 3 Sample at the expected peak position.

[0027] Figure 10 Selected Ion Chromatogram of 1,1,4a-Trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene in Example 3 Sample at the expected peak position time period. DETAILED DESCRIPTION

[0028] The application will be further described in conjunction with the drawings and examples.

[0029] Example 1 A method for detecting the authenticity and content of torreya grandis seed oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, comprising the following steps: A. Mix the vegetable oil sample uniformly, and take 1.00 g of the vegetable oil sample in a 25 ml test tube with a plug; B. Add 10 ml of chromatographically pure n-hexane to the test tube containing the vegetable oil sample, and vortex at high speed for 30 s; C. Add 3 ml of high-purity concentrated sulfuric acid to the mixed test tube, cover the plug, and vortex at high speed for 20 s; D. Place the vortexed test tube on the test tube rack and stand for 15 min, and then separate the layers to remove the fatty acids, pigments and impurities in the vegetable oil; E. Take 5 ml of the supernatant (n-hexane layer) after standing in another 25 ml test tube with a plug, and add 10 ml of 2% mass concentration of anhydrous sodium sulfate aqueous solution, and vortex at high speed for 20 s; F. Place the vortexed test tube on the test tube rack and stand for 15 min, and then separate the layers to remove the residual sulfuric acid in the n-hexane extract; G. Take 1 ml of the supernatant (n-hexane layer) after standing in F, pass through a 0.25 μm microporous filter membrane, and then put it into a sample bottle for gas chromatography-mass spectrometry detection and analysis; H. Establish a calibration curve for the percentage content of torreya grandis seed oil: take self-made and pressed torreya grandis seed oil, and supplement with soybean oil to 1 g if the weight is less than 1 g, and prepare torreya grandis seed oil with mass fractions of 0, 10%, 30%, 50%, 70%, and 100%, respectively, and analyze them simultaneously according to steps B-G, and establish a calibration curve for the mass percentage concentration of torreya grandis seed oil with the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene as the vertical coordinate and the mass percentage of torreya grandis seed oil as the horizontal coordinate; I. Whether 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is detected in the sample verifies whether the vegetable oil contains torreya grandis seed oil, and the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is brought into the calibration curve established in step H to calculate the mass percentage concentration of torreya grandis seed oil in the sample.

[0030] Chromatographic conditions of gas chromatography-mass spectrometer: Injection port: 250℃; Injection volume: 1 μl, no split injection; Chromatographic column: HP-5MS 5% Phenyl Methyl Silox (30 m x 0.25 mm, 0.25 μm); Carrier gas: He, flow rate 1 ml / min; Programmed temperature: 50℃ for 0.5min, 5℃ / min to 250℃ for 5min, 10℃ / min to 280℃ for 1min; Auxiliary heating zone (MSD transfer line): 280℃; Mass spectrometry acquisition mode: full scan (scan) for qualitative analysis, mass number 50.0~500; selected ion scan (sim) for quantitative analysis, mass number 159, residence time 50ms, mass number 255, residence time 100ms, mass number 270, residence time 50ms; Solvent delay: 3.0min; Ion source: 230℃; MS quadrupole rod: 150℃.

[0031] Detection results: Figure 2 、 Figure 3 、 Figure 4 are total ion chromatogram of the sample (containing "1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene" part), mass spectrum of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in the sample and selected ion spectrum of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in the sample, respectively.

[0032] Figure 2 、 Figure 3 It is shown that the sample contains torreya grandis seed oil, Figure 4 The peak area of the quantitative ion (255) is measured as 39621, and the calibration curve (y = 394.2 x + 1422, R 2 = 0.9998) obtained in the H step is brought in to calculate that the content of torreya grandis seed oil in the sample is 96.9%.

[0033] Conclusion: The content of torreya grandis seed oil in the sample is 96.9%±5%, which is pure torreya grandis seed oil.

[0034] Example 2 A method for detecting the authenticity and content of torreya grandis seed oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, comprising the following steps: A. Mix the plant oil sample uniformly, and take 1.00g of the plant oil sample in a 25ml test tube with a stopper; B. Add 10ml of chromatographically pure n-hexane to the test tube containing the plant oil sample, and vortex at high speed for 30s; C. Add 3 ml of concentrated sulfuric acid of analytical grade to the mixed test tube, cover the plug, and vortex at high speed for 20 s; D. Place the vortexed test tube on the test tube rack and stand for 15 min to remove the fatty acids, pigments, and impurities in the vegetable oil; E. Absorb 5 ml of the supernatant (n-hexane layer) after standing in another 25 ml test tube with a plug, and add 10 ml of 2% mass concentration of anhydrous sodium sulfate aqueous solution, vortex at high speed for 20 s; F. Place the vortexed test tube on the test tube rack and stand for 15 min to remove the residual sulfuric acid in the n-hexane extract; G. Absorb 1 ml of the supernatant (n-hexane layer) after standing in F, pass through a 0.25 μm microporous filter membrane, and then put it into a sample bottle for gas chromatography-mass spectrometry detection and analysis; H. Establish the calibration curve of the percentage content of torreya grandis seed oil: weigh the self-made and pressed torreya grandis seed oil, supplement it to 1 g with soybean oil if it is less than 1 g, and prepare torreya grandis seed oil with mass fractions of 0, 10%, 30%, 50%, 70%, and 100%, respectively, analyze them simultaneously according to steps B-G, take the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene as the vertical coordinate, and take the mass percentage of torreya grandis seed oil as the horizontal coordinate to establish the calibration curve of the mass percentage concentration of torreya grandis seed oil; I. Whether 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is detected verifies whether the vegetable oil contains torreya grandis seed oil, and the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is brought into the calibration curve established in step H to calculate the mass percentage concentration of torreya grandis seed oil in the sample.

[0035] Chromatographic conditions of gas chromatography-mass spectrometer: Inlet: 250℃; Injection volume: 1 μl, no split injection; Chromatographic column: HP-5MS 5% Phenyl Methyl Silox (30 m x 0.25 mm, 0.25 μm); Carrier gas: He, flow rate 1 ml / min; Programmed temperature: 50℃ for 0.5 min, 5℃ / min to 250℃ for 5 min, 10℃ / min to 280℃ for 1 min; Auxiliary heating zone (MSD transmission line): 280℃; Mass spectrometry acquisition mode: full scan (scan), mass number 50.0~500, for qualitative analysis; selected ion scan (sim), mass number 159, residence time 50ms, mass number 255, residence time 100ms, mass number 270, residence time 50ms, for quantitative analysis; Solvent delay: 3.0 min; Ion source: 230℃; MS quadrupole rod: 150℃.

[0036] Detection results: Figure 5 、 Figure 6 、 Figure 7 Respectively, the total ion chromatogram of the sample (containing the "1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene" part), the mass spectrum of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in the sample, and the selected ion spectrum of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in the sample.

[0037] Figure 5 、 Figure 6 Indicating that the sample contains torreya grandis seed oil, Figure 7 The quantitative ion (255) peak area is 14682, and the calibration curve (y = 394.2 x + 1422, R 2 = 0.9998) obtained by entering the H step is calculated to obtain the content of torreya grandis seed oil in the sample as 33.6%.

[0038] Conclusion: The content of torreya grandis seed oil in the sample is 33.6%±5%, and other oils are doped.

[0039] Example 3 A method for detecting the authenticity and content of torreya grandis seed oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, comprising the following steps: A. Mix the plant oil sample uniformly, and take 1.00g of the plant oil sample in a 25ml test tube with a plug; B. Add 10ml of chromatographically pure n-hexane to the test tube containing the plant oil sample and vortex at high speed for 30s; C. Add 3ml of high-purity concentrated sulfuric acid to the mixed test tube and vortex at high speed for 20s with the plug covered; D. Place the vortexed test tube on the test tube rack and stand for 15min, and then separate the layers to remove the fatty acids, pigments and impurities in the plant oil; E. Take 5 ml of the supernatant (n-hexane layer) after standing in another 25 ml test tube with a stopper, and add 10 ml of 2% mass concentration of anhydrous sodium sulfate aqueous solution, and vortex at high speed for 20 s; F. Place the vortexed test tube on the test tube rack for standing treatment for 15 min, and separate the layers to remove the residual sulfuric acid in the n-hexane extract; G. Take 1 ml of the supernatant (n-hexane layer) after F standing, pass through a 0.25 μm microporous filter membrane, and then put it into a sample bottle for gas chromatograph-mass spectrometer detection and analysis; H. Establish a calibration curve for the percentage content of torreya grandis seed oil: weigh the self-made and pressed torreya grandis seed oil, and supplement it with soybean oil to 1 g if it is less than 1 g, and prepare torreya grandis seed oil with mass fractions of 0, 10%, 30%, 50%, 70%, and 100%, respectively, and analyze them simultaneously according to steps B-G, and establish a calibration curve for the mass percentage concentration of torreya grandis seed oil with the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene as the vertical coordinate and the mass percentage of torreya grandis seed oil as the horizontal coordinate; I. Whether 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is detected verifies whether the plant oil contains torreya grandis seed oil, and the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is brought into the calibration curve established in step H to calculate the mass percentage concentration of torreya grandis seed oil in the sample.

[0040] Chromatographic conditions of gas chromatograph-mass spectrometer: Inlet: 250℃; Injection volume: 1 μl, no split injection; Chromatographic column: HP-5MS 5% Phenyl Methyl Silox (30 m x 0.25 mm, 0.25 μm); Carrier gas: He, flow rate 1 ml / min; Programmed temperature: 50℃ for 0.5 min, 5℃ / min to 250℃ for 5 min, 10℃ / min to 280℃ for 1 min; Auxiliary heating zone (MSD transmission line): 280℃; Mass spectrometry acquisition mode: full scan (scan), mass number 50.0~500, for qualitative analysis; selected ion scan (sim), mass number 159, residence time 50 ms, mass number 255, residence time 100 ms, mass number 270, residence time 50 ms, for quantitative analysis; Solvent delay: 3.0 min; Ion source: 230℃; MS quadrupole rod: 150℃.

[0041] Detection results: Figure 8 , Figure 9 , Figure 10 The total ion chromatogram of the sample (containing the peak position of "1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene") is Figure 9 The selected ion mass spectrum of the peak position of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in the sample of Example 3 and the selected ion chromatogram of the peak position time period of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene in the sample.

[0042] Figure 8 , Figure 9 , Figure 10 All show that "1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene" is not detected in the sample.

[0043] Conclusion: The sample is not Chinese torreya seed oil.

[0044] The above only describes specific embodiments of the present application, but the technical features of the present application are not limited thereto, and any person skilled in the art in the field of the present application can make equivalent changes and modifications within the scope of the patent application of the present application, which are all covered in the patent scope of the present application.

Claims

1. A method for detecting the authenticity and content of torreya grandis seed oil based on the characteristics of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, characterized in that, The method comprises the following steps: A. Mix the vegetable oil sample uniformly, and weigh the vegetable oil sample into a test tube with a plug; B. Add n-hexane to dissolve the vegetable oil in the test tube with the vegetable oil sample, cover the plug, and mix uniformly at high speed; C. Add concentrated sulfuric acid to the mixed test tube, cover the plug, and mix uniformly at high speed; D. Place the vortexed test tube on a test tube rack for static treatment and stratification to remove fatty acids, pigments and impurities in the vegetable oil; E. Absorb the supernatant after static treatment into another test tube with a plug, add an aqueous sodium sulfate solution, cover the plug, and vortex at high speed; F. Place the vortexed test tube on a test tube rack for static treatment and stratification to remove residual sulfuric acid in the n-hexane extract; G. Absorb 1 ml of the supernatant after static treatment in F, pass through a microporous filter membrane, and then put into a sample bottle, and then detect and analyze by a gas chromatograph-mass spectrometer; H. Establish a calibration curve of the percentage content of torreya grandis seed oil: weigh the self-made and pressed torreya grandis seed oil, supplement the vegetable oil to 1 g if less than 1 g, prepare torreya grandis seed oil with mass fractions of 0, 10%, 30%, 50%, 70%, and 100%, respectively, analyze simultaneously according to steps B-G, take the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene as the vertical coordinate, and take the mass percentage of torreya grandis seed oil as the horizontal coordinate to establish a calibration curve of the mass percentage concentration of torreya grandis seed oil; I. Whether 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is detected verifies whether the vegetable oil contains torreya grandis seed oil, and the peak area of 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene is brought into the calibration curve established in step H to calculate the mass percentage concentration of torreya grandis seed oil in the sample.

2. The method according to claim 1 for detecting the authenticity and content of Japanese nut oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene characteristics, characterized in that, In step A, 0.5-1.5 g of the vegetable oil sample is weighed into a 25 ml test tube with a plug, and preferably the mass of the vegetable oil sample is 1.00 g.

3. The method according to claim 1 for detecting the authenticity and content of Japanese nut oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene characteristics, characterized in that, In step B, the n-hexane is chromatographically pure n-hexane, 10 ml of n-hexane is added, and the vortexing time is 10-30 s, and preferably the vortexing time is 30 s.

4. The method according to claim 1 for detecting the authenticity and content of Japanese nut oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene characteristics, characterized in that, In step C, the concentrated sulfuric acid is premium grade concentrated sulfuric acid, the volume is 3-5 ml, and preferably 3 ml, the plug is covered, and the vortexing is performed at high speed for 20 s.

5. The method according to claim 1 for detecting the authenticity and content of Japanese nut oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene characteristics, characterized in that, In steps D and F, the stratification time for static treatment is 10-20 min, and preferably 15 min.

6. The method according to claim 1 for detecting the authenticity and content of Japanese nut oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene characteristics, characterized in that, In step E, the volume of the supernatant is 3-5 ml, and preferably 5 ml; the aqueous sodium sulfate solution added is a 2% mass concentration of anhydrous sodium sulfate aqueous solution of 10 ml, and the vortexing is performed at high speed for 20 s.

7. The method according to claim 1 for detecting the authenticity and content of Japanese nut oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a- octahydrophenanthrene characteristics, characterized in that, In step G, 1-2 ml of the supernatant is absorbed, and preferably 1 ml; the pore size of the microporous filter membrane is 0.25 μm.

8. A method for detecting the authenticity and content of torreya grandis seed oil based on 1,1,4a-trimethyl-7-isopropyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene according to any one of claims 1-7, characterized in that, In step H, the supplementary vegetable oil is soybean oil.

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