Method for analyzing volatile components of roxburgh rose flavor substances and application of method

Abstract

The invention discloses a roxburgh rose flavor substance analysis method and application thereof, and belongs to the technical field of traditional Chinese medicine analysis. Comprising the following steps: (1) extracting a roxburgh rose sample by adopting a volatile oil extraction method to obtain a test solution; (2) detecting the roxburgh rose flavor substances in the test solution by adopting a gas chromatography-mass spectrometry method; in the step (1), the sampling amount of the rosa roxburghii tratt extract is that the mass of solid matters in the rosa roxburghii tratt extract is 20-30g, and the conditions of gas chromatography and mass spectrometry in the step (2) are described in the specification. According to the analysis method provided by the invention, the flavor substance fingerprints of the roxburgh rose clear juice and the roxburgh rose concentrated juice are constructed, volatile components in the roxburgh rose flavor substances are determined, the established method meets methodological requirements, a standard basis is provided for detection of the roxburgh rose extract flavor substances, and development of the roxburgh rose related flavor substances is facilitated.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine analysis, and particularly relates to a method for analyzing volatile components of rose apple flavor substances and application thereof. BACKGROUND

[0002] Rose apple (Rosa roxburghii Tratt), also known as raspberry fruit and Wenxian fruit, belongs to a kind of shrub of Rosaceae Rosaceae, and is the fruit of a kind of perennial deciduous shrub, which is rich in vitamin C, flavonoids, polyphenols, superoxide dismutase (SOD), polysaccharides and other substances, and has high edible and medicinal value. In addition to fresh rose apple, there are many processed products of rose apple on the market, such as rose apple preserved fruit, rose apple powder, rose apple dry, rose apple juice and rose apple concentrated juice. At present, the production of rose apple, from cultivation, management and protection, picking, processing, sales, the monitoring method of effective components and related flavor substance components in the production process of raw materials, intermediate products and finished products has problems such as complicated determination process, unobvious target components and single index components. For example, Chinese patent application CN118584001A discloses a method for preparing rose apple root extract, a detection method and a method for constructing a characteristic spectrum, the rose apple root extract is obtained by two times of water decoction and filtration of rose apple root decoction pieces, and the decoction liquid is concentrated and dried to obtain the rose apple root extract. The detection method of the rose apple root extract is established by taking the content of ellagic acid as an index component. However, since only ellagic acid is used as an index component, the detection component is single, and the overall quality of the product cannot be comprehensively reflected. Chinese patent application CN113533553A discloses a detection method of rose apple and its products and a method for establishing a rose apple component database. Based on LC-MS / MS technology, real-time, rapid and economical detection of rose apple sample components is realized. In addition, the existing data resources are compared and identified with the LC-MS / MS results. The obtained identification results are established into a rose apple database, and different rose apple samples are used to verify the database, which simultaneously realizes the quality analysis of each rose apple sample, helps the control of the quality of rose apple products, and is beneficial to the further exploration of the efficacy of rose apple. However, the above-mentioned patent application includes more than 120 components, the target components are not outstanding, and the above-mentioned patent application does not give which components are the index components for identifying the flavor of rose apple. Chinese patent CN115343429B discloses a detection method of volatile flavor substances of spice oil based on PCA analysis technology and application, and Heracles II ultrafast gas phase electronic nose is used to monitor the volatile flavor substances in each process of the processing of pepper seasoning oil. However, the detection method is not suitable for the detection of rose apple flavor substances.

[0003] Roselle lacks clear flavor markers, making it difficult to scientifically quantify the quality differences of roselle from different producing areas and varieties, affecting standardized production. Meanwhile, beverages, preserved fruits and other products need to be manually flavored to make up for processing losses, but due to the lack of basic data, relying on trial and error, the cost is high and the efficiency is low. SUMMARY

[0004] In order to solve the problems in the prior art, the present application provides a method for analyzing volatile components of roselle flavor substances and its application. According to the characteristics of roselle extract and flavor substances, the method for preparing the test sample is optimized, the fingerprint method of the flavor substances of the roselle extract is established, the related volatile index components of the flavor substances in the roselle extract are determined, and the method is verified. At the same time, the fingerprint of the flavor substances of the roselle product is established.

[0005] The first aspect of the present application relates to a method for analyzing volatile components of roselle flavor substances, which comprises the following steps: (1) treating the roselle extract by volatile oil extraction method to obtain a test sample solution; (2) detecting the volatile components of the roselle flavor substances in the test sample solution by gas chromatography-mass spectrometry (GC-MS) method; wherein, In step (1), the sampling amount of the roselle extract is 20-30g of solid mass; In step (2), the temperature programming conditions of the gas chromatograph are: the initial temperature is 50℃, which is increased to 250℃ at a rate of 3℃ / min, and maintained for 5min; the conditions of the mass spectrometer are: detected by triple quadrupole mass spectrometer, the ion source is electron impact source, the ion source temperature is 240±20℃, the collision gas is argon, the mass spectrometer transmission line temperature is 240℃, the mass spectrometer monitoring mode is SCAN full scan detection mode, the solvent delay time is 0-7min, the MS program: 7min filament on, 10min filament off, 13min filament on.

[0006] Further preferably, in step (1), the roselle extract is roselle clear juice or roselle concentrated juice.

[0007] Further preferably, the solid mass is 20g.

[0008] Preferably, the preparation step of the test sample solution in step (1) is: accurately weighing a proper amount of roselle sample, adding water and glass beads to make the solid concentration of the roselle sample 2-3g / 100mL, connecting the volatile oil tester, adding water and ethyl acetate from the upper end of the tester, connecting the reflux condenser, micro-boiling extraction, collecting the ethyl acetate layer, and constant volume with ethyl acetate.

[0009] Further preferably, the time of the micro-boiling extraction is 3-5 h, the added amount of ethyl acetate is 2 mL, and the final volume of the ethyl acetate constant volume is 2 mL.

[0010] Preferably, the gas chromatography conditions in step (2) are as follows: SH-624 capillary column, high-purity helium as the carrier gas, carrier gas flow rate of 1 mL / min, injection port temperature of 240±20℃, split injection, split ratio of 2:1, and injection volume of 1 μL.

[0011] Further preferably, the injection port temperature is 240℃.

[0012] Preferably, in step (2), the ion source temperature is 250℃.

[0013] The second aspect of the present application relates to the use of the above-mentioned analysis method in the construction of a fingerprint of volatile components of a flavor substance of a roxburgh rose product.

[0014] Preferably, the relevant index components in the fingerprint are dimethyl sulfide, dimethyl butyraldehyde, butyric acid, leaf alcohol, furfural, hexanol, leaf alcohol acetate, hexanoic acid, linalool, octanoic acid, α-terpineol and α-ionol.

[0015] Compared with the prior art, the present application has the following beneficial effects: (1) the fingerprint method of flavor substances of roxburgh rose extract is established through optimization of the test sample preparation method; (2) the method is verified for repeatability, stability and durability, and the results show that the established method meets the methodological requirements; (3) the fingerprint method of 12 components of roxburgh rose clear juice and roxburgh rose concentrated juice is established, and methodological study and similarity evaluation are performed, thereby providing a standard basis for roxburgh rose extract and effectively controlling the quality of raw materials and volatile components of flavor substances of roxburgh rose extract; (4) 37 common substances of roxburgh rose extract are explored by using gas chromatography-mass spectrometry (GC-MS) and NIST spectral library, and the flavor and health function of roxburgh rose products are improved. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 : the SCAN full scan spectrum of the roxburgh rose extract test sample prepared by the solvent extraction method in experimental example 1; Figure 2 : the SCAN full scan spectrum of the roxburgh rose extract test sample prepared by the static headspace method in experimental example 1; Figure 3 : the SCAN full scan spectrum of the roxburgh rose extract test sample prepared by the volatile oil extraction method in experimental example 1, wherein S1 is roxburgh rose clear juice, and S2 is roxburgh rose concentrated juice; Figure 4 : the spectrum after similarity matching of the repeatability solution in experimental example 2; Figure 5: The chromatogram of the stability solution after similarity matching of the experimental example 2, wherein A-F represent the chromatogram of the test solution at 0h, 6h, 12h, 24h, 36h and 48h respectively; Figure 6 : The full scan overlay of the gas chromatography injection port temperature investigation of the experimental example 2, wherein A-C represent the chromatogram of the injection port temperature of 220℃, 240℃ and 260℃ respectively; Figure 7 : The full scan overlay of the ion source temperature investigation of the mass spectrometry of the experimental example 2, wherein A-C represent the chromatogram of the ion source temperature of 220℃, 240℃ and 260℃ respectively; Figure 8 : The flavor substance fingerprint of the Rosa roxburghii clear juice; Figure 9 : The total ion current chromatogram of the mixed reference solution; Figure 10 : The total ion current chromatogram of the test solution of the Rosa roxburghii clear juice; Figure 11 : The flavor substance fingerprint of the Rosa roxburghii concentrated juice; Figure 12 : The total ion current chromatogram of the test solution of the Rosa roxburghii concentrated juice; Figure 13 : The fingerprint of the Rosa roxburghii extract; Note: Figure 1 0-6min is solvent delay, Figure 3 0-3.5min is solvent delay, Figures 4-13 0-7min is solvent delay, the chromatogram is not shown, which does not affect the confirmation of the chromatogram of the test substance in the test solution; in Figure 9 , Figure 10 , Figure 12 , peak 1 is dimethyl sulfide, peak 2 is dimethyl butyraldehyde, peak 6 is butyric acid, peak 7 is leaf alcohol, peak 8 is furfural, peak 9 is hexanol, peak 11 is ethyl acetic acid leaf alcohol ester, peak 12 is hexanoic acid, peak 15 is linalool, peak 16 is octanoic acid, peak 17 is alpha-terpineol, and peak 20 is alpha-ionol. DETAILED DESCRIPTION

[0017] The present application is described below by examples to make the technical scheme of the present application easier to understand and grasp, but the present application is not limited to this. The experimental methods described in the following examples are all conventional methods, unless otherwise specified; the medicinal materials and reagents described in the following examples are all commercially available, unless otherwise specified; and the performance of products from different sources does not have a significant impact.

[0018] When the embodiments give numerical ranges, it is to be understood that unless the application specifically states otherwise, the endpoints of the ranges and any values between the endpoints are included. Unless defined otherwise, all technical and scientific terms used in the application have the same meaning as is commonly understood by one of ordinary skill in the art to which this application belongs.

[0019] The various chemical reagents and raw materials used in the embodiments of the application are obtained through conventional commercial channels unless otherwise specified, and the products from different manufacturers do not have a significant effect on the results.

[0020] The list of equipment used in the following experiments is shown in Table 1, the information of the reference substance is shown in Table 2, and the information of the roxburgh rose sample is shown in Table 3.

[0021] Table 1: List of equipment

[0022] Table 2: Information of reference substance

[0023] Table 3: Information of sample

[0024] The following experimental conditions are used in the following experiments unless otherwise specified: Chromatographic conditions: SH-624 capillary column (60 m x 0.32 mm x 1.8 μm). The carrier gas is high-purity helium (He), and the carrier gas flow rate is 1 mL / min. The programmed temperature is 50°C initially, and then increased to 250°C at a rate of 3°C / min, and maintained for 5 min. The injection port temperature is 240°C. Split injection is used, and the split ratio is 2:1. The injection volume is 1 μL.

[0025] Mass spectrometry conditions: detected by triple quadrupole tandem mass spectrometer; the ion source is electron impact source (EI), and the ion source temperature is 250°C. The collision gas is argon. The mass spectrometry transmission line temperature is 240°C. The mass spectrometry monitoring mode is SCAN full scan detection mode. The solvent delay time is 0-7 min. The MS program is 7 min with the filament on, 10 min with the filament off, and 13 min with the filament on.

[0026] Experimental Example 1: Preparation of test solution and investigation of extraction method The solvent extraction method, static headspace method and volatile oil extraction method were used to process roxburgh rose extract (roxburgh rose concentrated juice with sample number QT23110037) to obtain different test solutions, and the specific processing process is as follows: Solvent extraction method: 200 mL of concentrated Malus micromalus juice numbered QT23110037 and 10 μL of cyclohexanone were measured in a 500 mL separatory funnel, 50 mL, 40 mL and 30 mL of dichloromethane were used to extract three times respectively, and then the extract was combined, 5 g of anhydrous sodium sulfate was added to the extract, after water removal, filtration, the filtrate was concentrated to 500 μL using nitrogen as the test sample solution.

[0027] Static headspace method: 2 g of sodium chloride was weighed and added to a 20 mL headspace bottle, 5 mL of concentrated Malus micromalus juice numbered QT23110037 was accurately measured and added to the headspace bottle, sealed with a cover, placed in a headspace sample plate, and the headspace bottle was balanced at a temperature of 80°C for 30 min, the quantitative ring temperature was 110°C, and the transmission line temperature was 115°C. The test sample was obtained.

[0028] Volatile oil extraction method: (1) A certain amount of concentrated Malus micromalus juice numbered QT23110037 (equivalent to 20 g of solid content) was precisely weighed and placed in a 1000 mL flask, 300 mL of water and a few glass beads were added, mixed by shaking, and then connected to a volatile oil detector. Add water to the upper end of the detector to fill the scale part, and overflow into the flask. Then use a pipette to add 2 mL of ethyl acetate, and then connect a reflux condenser. Slowly heat to boiling in an electric heating jacket and maintain a slight boil for about 4 hours, stop heating. Place for 1 hour. Collect the ethyl acetate layer, filter with anhydrous sodium sulfate, and then place in a 2 mL volumetric flask. Add ethyl acetate to the mark to obtain the test sample solution of the concentrated Malus micromalus juice.

[0029] (2) Take Malus micromalus juice numbered QT23110079, and prepare the test sample solution of Malus micromalus juice in the same way as (1) above.

[0030] The test sample solutions prepared by the above three methods were detected by GC-MS, and the GC-MS conditions were as follows: Chromatographic conditions: SH-624 capillary column (60 m x 0.32 mm x 1.8 μm). The carrier gas is high-purity helium (He), and the carrier gas flow rate is 1 mL / min. The temperature program is as follows: the initial temperature is 50°C, which is increased to 250°C at a rate of 3°C / min, and maintained for 5 min. The injection port temperature is 240°C. Split injection, split ratio 2:1. Injection volume: 1 μL.

[0031] Mass spectrometry conditions: detected by triple quadrupole mass spectrometer; ion source is electron impact source (EI), ion source temperature: 250°C. Collision gas is argon. Mass spectrometry transmission line temperature: 240°C. Mass spectrometry monitoring mode: SCAN full scan detection mode. Solvent delay time: 0-7 min. MS program: 7 min filament on, 10 min filament off, 13 min filament on.

[0032] The chromatograms of the three test samples are shown in Figures 1-3

[0033] From Figure 1 it can be seen that the number of signal peaks scanned after the test sample solution prepared by solvent extraction method is injected and analyzed is less, which does not meet the expected requirements. From Figure 2 it can be seen that the number of signal peaks scanned after the test sample solution prepared by solvent extraction method is injected and analyzed is less, which does not meet the expected requirements. From Figure 3 it can be seen that the number of signal peaks scanned after the test sample solution prepared by solvent extraction method is injected and analyzed is less, which does not meet the expected requirements. From

[0034] Therefore, the test sample solution is prepared by the volatile oil extraction method.

[0035] Experimental Example 2: Methodology verification of the analysis method The sample No. QT23110037 concentrated juice of Rosa roxburghii Tratt was selected to verify the repeatability, stability and durability, and the results showed that the established method met the methodological requirements and could be used for the construction of the fingerprint of Rosa roxburghii Tratt extract.

[0036] (1) Repeatability Six portions of concentrated juice of Rosa roxburghii Tratt were taken, and six test sample solutions were prepared according to the volatile oil extraction method in Experimental Example 1. The samples were analyzed by injection according to the GC-MS conditions in Experimental Example 1, and the similarity of the obtained chromatograms was evaluated by the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 version). The results are shown in Figure 4 and Table 4.

[0037] Table 4: Similarity of repeatability solutions

[0038] According to the calculation of the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System, the minimum similarity of the fingerprint of the six repeatedly prepared test sample solutions was 0.997, which met the requirements (≥0.90). It showed that the repeatability of the method was good.

[0039] (2) Stability The concentrated juice of Rosa roxburghii Tratt was taken, and one test sample solution was prepared according to the volatile oil extraction method in Experimental Example 1. The samples were analyzed by injection according to the chromatographic conditions in Experimental Example 1 at 0 h, 6 h, 12 h, 24 h, 36 h and 48 h, and the similarity of the obtained chromatograms was evaluated by the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 version). The results are shown in Figure 5 and Table 5.

[0040] Table 5: Stability results of the method

[0041] ​The similarity of the fingerprint chromatogram of the test solution at the 6 observation time points was calculated by the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicines, and the minimum similarity was 0.980, which met the requirement (≥0.90). It indicated that the test solution had good stability within 48 h.

[0042] (3) Durability ① Injection port temperature The concentrated juice of Rosa roxburghii was taken, and 1 portion of the test solution was prepared according to the volatile oil extraction method in Experimental Example 1. The injection was performed for 3 times. Except that the injection port temperature was 220 ℃, 240 ℃ and 260 ℃ respectively, the other conditions were the same as the chromatographic conditions in Experimental Example 1. The similarity of the obtained chromatograms was evaluated by the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicines (2012 version). The results are shown in Table 6. Figure 6

[0043] Table 6: Comparison of similarity under different injection port temperature conditions

[0044] The similarity of the fingerprint chromatogram of the test solution at the 6 observation time points was calculated by the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicines, and the minimum similarity was 0.980, which met the requirement (≥0.90). It indicated that the test solution had good stability within 48 h.

[0045] ② Ion source temperature The concentrated juice of Rosa roxburghii was taken, and 1 portion of the test solution was prepared according to the volatile oil extraction method in Experimental Example 1. The injection was performed for 3 times. Except that the ion source temperature was 220 ℃, 240 ℃ and 260 ℃ respectively, the other conditions were the same as the chromatographic conditions in Experimental Example 1. The similarity of the obtained chromatograms was evaluated by the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicines (2012 version). The results are shown in Table 7. Figure 7

[0046] Table 7: Comparison of similarity under different ion source temperature conditions

[0047] The similarity of the fingerprint chromatogram of the test solution at the 6 observation time points was calculated by the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicines, and the minimum similarity was 0.980, which met the requirement (≥0.90). It indicated that the test solution had good stability within 48 h.

[0048] Experimental Example 3: Establishment of fingerprint chromatogram of Rosa roxburghii juice (1) Solid content determination method: 5 g of sample was taken and placed on a constant weight aluminum pan of a water determination instrument. When the sample was dried to constant weight, the mass was read and calculated to obtain the solid content. The solid content of 16 batches of Rosa roxburghii juice is shown in Table 8.

[0049] Table 8: Solid content of 16 batches of Rosa roxburghii juice​​

[0050] (2) Similarity comparison of 16 batches of roxburgh rose clear juice fingerprint Take 16 batches of roxburgh rose clear juice, respectively, after determining the solid content, prepare the test solution according to the volatile oil extraction method in experimental example 1, sample analysis according to the chromatographic condition in experimental example 1, the obtained chromatogram is introduced into traditional Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version), set the time window as 0.20, automatically match to obtain the roxburgh rose clear juice control fingerprint. The results are shown in Figure 8 and table 9.

[0051] Table 9: Similarity comparison of 16 batches of roxburgh rose clear juice fingerprint

[0052] The minimum similarity of roxburgh rose clear juice fingerprint is 0.904, which meets the requirements (≥0.90).

[0053] (3) Common peak identification Preparation of mixed control solution: precisely weigh 10 mg of each of the 12 kinds of control samples in table 2 into a 10 mL volumetric flask, add ethyl acetate for dissolution, and prepare 12 kinds of mixed control sample stock solution. Precisely take 200 μL of mixed control sample stock solution into a 10 mL volumetric flask, and dilute to the mark with ethyl acetate, mix well, which is the mixed control solution.

[0054] Through comparison with the control sample spectrum, 21 common peaks of roxburgh rose clear juice fingerprint are identified, as shown in Figure 9 . Among them, peak 1 (dimethyl sulfide), peak 2 (dimethyl butanal), peak 6 (butyric acid), peak 7 (leaf alcohol), peak 8 (furfural), peak 9 (hexanol), peak 11 (acetyl leaf alcohol ester), peak 12 (hexanoic acid), peak 15 (linalool), peak 16 (octanoic acid), peak 17 (α-terpineol), peak 20 (α-ionol).

[0055] Experimental example 4: establishment of roxburgh rose concentrated juice fingerprint (1) Solid content determination method Take 12 batches of roxburgh rose concentrated juice, respectively, according to the solid content determination method in experimental example 3, the results are shown in table 10.

[0056] Table 10: Solid content of 12 batches of roxburgh rose concentrated juice

[0057] (2) Similarity comparison of 12 batches of roxburgh rose concentrated juice fingerprint The sample solution was prepared according to the volatile oil extraction method in Experimental Example 1, and was analyzed by injection according to the chromatographic conditions in Experimental Example 1. The obtained chromatogram was introduced into the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 version), and the time window was set to 0.20. The control fingerprint of the concentrated Malus sieversii juice was automatically matched. The results are shown in Table 11. Figure 10

[0058] Table 11: Similarity comparison of 12 batches of concentrated Malus sieversii juice fingerprint

[0059] The minimum similarity of the concentrated Malus sieversii juice fingerprint was 0.924, which met the requirements (≥0.90).

[0060] (3) Identification of common peaks The mixed control solution was prepared according to Experimental Example 3. Through comparison with the control fingerprint, there were 21 common peaks in the concentrated Malus sieversii juice fingerprint, as shown in Table 11. Figure 11 Among them, peak 1 (dimethyl sulfide), peak 2 (dimethyl butanal), peak 6 (butyric acid), peak 7 (leaf alcohol), peak 8 (furfural), peak 9 (hexanol), peak 11 (acetic acid leaf alcohol ester), peak 12 (hexanoic acid), peak 15 (linalool), peak 16 (octanoic acid), peak 17 (α-terpineol), and peak 20 (α-ionol) Experimental Example 5: Establishment of Malus sieversii extract fingerprint The fingerprints of 16 batches of clear juice and 12 batches of concentrated juice were placed together, and the similarity results are shown in Table 12-1, Table 12-2 (due to the large amount of data, the font is too small in one table, so the data is divided into two tables for display) and Figure 12 .

[0061] Table 12-1: Similarity comparison of 16 batches of clear juice and 12 batches of concentrated juice fingerprint

[0062] Table 12-2: Similarity comparison of 16 batches of clear juice and 12 batches of concentrated juice fingerprint

[0063] The similarity of the Malus sieversii clear juice control fingerprint, the Malus sieversii concentrated juice control fingerprint, and the Malus sieversii extract fingerprint was compared, and the results are shown in the following table: Table 13: Similarity comparison of fingerprints

[0064] ​Conclusion: The similarity between the 28 batches of Malus hallings extract fitting fingerprint is the smallest, which is 0.765, and the similarity between the matching fingerprint is the smallest, which is 0.935. The similarity between the Malus hallings clear juice control fingerprint, Malus hallings concentrated juice control fingerprint and Malus hallings extract fingerprint is the smallest, which is 0.948, meeting the requirements (≥0.90). Therefore, the Malus hallings clear juice control fingerprint and Malus hallings concentrated juice control fingerprint can be fitted into one Malus hallings extract fingerprint, which can be used for the analysis of Malus hallings extract.

[0065] Experimental Example 6: Study on flavor substances of Malus hallings extract The common substances of Malus hallings extract were explored by gas chromatography-mass spectrometry (GC-MS) and NIST spectral library, and analyzed.

[0066] Through literature search, 83 flavor substances of Malus hallings fruit have been identified under different processing methods and extraction processes, and the specific list of substances is shown in Table 14.

[0067] Table 14: Flavor substances of Malus hallings

[0068]

[0069]

[0070] Screening of common substances of Malus hallings extract: Take 8 batches of Malus hallings extract numbered as QT23110037, QT23110038, QT23110102-QT23110106 and QT23110079, respectively determine the solid content, then prepare the test solution according to the volatile oil extraction method in Experimental Example 1, analyze by the chromatographic conditions in Experimental Example 1, and match the similarity degree in NIST spectral library. The substances with similarity degree in the top 70 are shown in Table 15: Table 15: NIST spectral library qualitative results of concentrated juice

[0071]

[0072]

[0073]

[0074]

[0075] Screening of common substances of 8 batches of Malus hallings extract in Table 15 showed that there were 37 common substances, which are shown in Table 16.

[0076] Table 16: Common substances of Rosa roxburghii extract

[0077] Relationship between flavor substances and fingerprint of Rosa roxburghii: There are 4 common substances between Rosa roxburghii extract (Table 17) and literature search substances (Table 15), which are ethyl butyrate (CAS No. 105-54-4), dimethyl sulfide (peak 1, CAS No. 75-18-3), 2-methylbutanal (peak 2, CAS No. 96-17-3), and 3-methylbutanal (CAS No. 590-86-3). Among them, dimethyl sulfide is peak 1 in the fingerprint of Rosa roxburghii, and 2-methylbutanal is peak 2 in the fingerprint of Rosa roxburghii.

[0078] The volatile oil extraction method was used to establish the fingerprint of volatile components of Rosa roxburghii extract by GC-MS full scan, and the flavor substances of Rosa roxburghii extract were studied by comparing the similarity of NIST spectrum library, and 37 common substances were obtained. Four common substances were determined by analysis, among which peak 1 and peak 2 in the fingerprint were identified as flavor substances of Rosa roxburghii.

[0079] Finally, it should be noted that the above content is only used to illustrate the technical solutions of the present application, and is not a limitation on the protection scope of the present application. Simple modifications or equivalent replacements of the technical solutions of the present application made by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.

Claims

1. A method for analyzing volatile components of a Malus baccata flavoring substance, characterized by, The analysis method comprises the following steps: (1) treating the rosehip extract by a volatile oil extraction method to obtain a test sample solution; (2) detecting the volatile flavor components in the test sample solution by a gas chromatography-mass spectrometry method; wherein, In step (1), the sampling amount of the rosehip extract is 20-30 g of solid content; In step (2), the temperature programming condition of the gas chromatography is that the initial temperature is 50℃, and is increased to 250℃ at a rate of 3℃ / min and maintained for 5 min; the condition of the mass spectrometry is that a triple quadrupole tandem mass spectrometer is used for detection, the ion source is an electron impact source, the ion source temperature is 240±20℃, the collision gas is argon, the mass spectrometry transmission line temperature is 240℃, the mass spectrometry monitoring mode is a SCAN full-scan detection mode, the solvent delay time is 0-7 min, the MS program is that the filament is turned on for 7 min, turned off for 10 min, and turned on for 13 min.

2. The analysis method according to claim 1, characterized in that The solid content is 20 g.

3. The analysis method of claim 1, wherein, In step (1), the preparation of the test sample solution is as follows: a proper amount of rosehip sample is accurately weighed, water and glass beads are added to make the solid content of the rosehip sample 2-3 g / 100 mL, a volatile oil tester is connected, water and ethyl acetate are added to the upper end of the tester, a reflux condenser tube is connected, micro-boiling extraction is performed, the ethyl acetate layer is collected, and the ethyl acetate is made up to volume, and the test sample solution is obtained.

4. The analysis method according to claim 3, characterized in that, The micro-boiling extraction time is 3-5 h, the amount of ethyl acetate added is 2 mL, and the final volume of the ethyl acetate made up to volume is 2 mL.

5. The analysis method of claim 1, wherein, In step (2), the condition of the gas chromatography is that a SH-624 capillary column is used, high-purity helium is used as the carrier gas, the carrier gas flow rate is 1 mL / min, the injection port temperature is 240±20℃, split injection is used, the split ratio is 2:1, and the injection volume is 1 μL.

6. The analysis method according to claim 5, characterized in that, The injection port temperature is 240℃.

7. The analysis method of claim 1, wherein, In step (2), the ion source temperature is 250℃.

8. The use of the analysis method according to any one of claims 1-7 in the construction of a fingerprint of volatile flavor components of rosehip products.

9. Use according to claim 8, characterized in that, The relevant index components in the fingerprint are dimethyl sulfide, dimethyl butyraldehyde, butyric acid, leaf alcohol, furfural, hexanol, leaf alcohol acetate, hexanoic acid, linalool, octanoic acid, alpha-terpineol and alpha-ionol.

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