Cubilose origin traceability method based on high-field nuclear magnetic resonance technology and its application in quality monitoring

Abstract

The application provides a bird's nest origin tracing method based on high-field nuclear magnetic resonance technology, comprising the following steps: taking bird's nests from different origins and a to-be-tested substance, performing extraction treatment, and preparing a to-be-tested sample; performing pretreatment on the to-be-tested sample before high-field nuclear magnetic resonance detection, collecting detection data; performing multivariate statistical analysis on the detection data; wherein the extraction treatment comprises the following steps: soaking the bird's nests from different origins and the to-be-tested substance, mixing them with artificial saliva respectively, performing first water bath heating, and preparing a first mixed solution; mixing the first mixed solution with artificial gastric juice, performing second water bath heating, and then performing enzyme inactivation; the second water bath heating conditions comprise: a temperature of 37 DEG C + / - 1 DEG C, and a time of 3h+ / -0.5h; the high-field nuclear magnetic resonance detection comprises the following conditions: a frequency spectrum width of 12kHz+ / -2kHz, and an accumulated scanning number of 64 times+ / -8 times. The method is efficient, accurate, convenient, environmentally friendly, and low in cost.

Description

Technical Field

[0001] This invention relates to the field of bird's nest quality control technology, and in particular to a method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology and its application in quality monitoring. Background Technology

[0002] Bird's nest is the nest built by swiftlets and other species of swiftlets, mainly produced in Southeast Asian countries and regions such as Malaysia and Indonesia. It has long been regarded as a precious traditional Chinese medicine and a rare food. To effectively regulate the quality of bird's nest, research on the identification of genuine bird's nest has been continuously developing and innovating.

[0003] The purpose of bird's nest origin traceability is to determine the actual place of origin of bird's nest through rapid and effective traceability technology, thereby enabling an objective evaluation of the quality and price of bird's nest and preventing the sale of inferior products. Currently, the technologies used in food traceability mainly include IoT and label traceability, isotope traceability, near-infrared spectroscopy traceability, mineral element traceability, DNA traceability, and organic component traceability. IoT and label traceability primarily uses tattoos, labels, animal ID cards, and RFID to identify the origin of products. While these methods are intuitive, they are prone to problems such as lost labels, incorrect records, incomplete information, and tampering with product information, making it impossible to obtain the true origin information and introducing significant human error into traceability. In addition, chemical methods can be used to identify the product's origin. Mass spectrometry, spectroscopy, chromatography, and other techniques are used to analyze the product, combined with chemometrics, to accurately identify the true origin of the product. Chemical component analysis is considered a relatively effective and accurate traceability method with good development prospects. In the food industry, common chemical composition analysis methods include isotope tracing (IRMS), near-infrared spectroscopy tracing (near-infrared spectrometer), mineral element tracing (ICP-MS), and chromatographic analysis tracing (gas chromatography, liquid chromatography-mass spectrometry). Regarding mineral element tracing, traditional techniques using inductively coupled plasma mass spectrometry (ICP-MS) to determine elements in bird's nest require sample digestion, which involves the use of toxic and harmful reagents such as nitric acid and hydrogen peroxide, posing hazards to the environment and laboratory personnel. The testing process takes approximately 1 to 2 days. In the biological field, DNA tracing technology is a relatively common tracing method. Traditional techniques use DNA source identification to distinguish bird's nests from different origins. The sample pretreatment steps require extracting total DNA, followed by PCR amplification and sequencing. The entire sample processing is quite complex, involving more than a dozen reagents and equipment. Furthermore, the PCR amplification process is susceptible to factors such as temperature and cleanliness, which can easily lead to DNA aerosol contamination. This requires high standards for the environment and personnel's operating techniques, and the testing time is relatively long, taking about 2 to 3 days.

[0004] Currently, there are no methods or studies on applying nuclear magnetic resonance (NMR) technology to trace the origin of bird's nest, and there are no reports on applying NMR technology to trace the origin of food in the food industry. Summary of the Invention

[0005] Based on this, the present invention provides a method for tracing the origin of bird's nests based on high-field nuclear magnetic resonance technology, which provides a more efficient, accurate, convenient, environmentally friendly and low-cost method for identifying the origin of bird's nests.

[0006] The present invention is achieved through the following technical solution.

[0007] A method for tracing the origin of bird's nests based on high-field nuclear magnetic resonance technology includes the following steps:

[0008] Bird's nests from different origins and the test substance were extracted, processed, and used to prepare the test samples;

[0009] The sample to be tested is pretreated and then subjected to high-field nuclear magnetic resonance detection to collect detection data.

[0010] The detection data were subjected to multivariate statistical analysis.

[0011] The extraction process includes the following steps: soaking the bird's nests from different origins and the test substance, mixing them separately with artificial saliva, heating them in a water bath for the first time to prepare a first mixture; mixing the first mixture with artificial gastric juice, heating it in a water bath for the second time, and then performing enzyme inactivation; the conditions for the second water bath heating include: temperature of 37℃±1℃ and time of 3h±0.5h.

[0012] High-field NMR detection includes the following conditions: spectral width of 12 kHz and cumulative scan count of 64.

[0013] In one embodiment, the pretreatment includes the following steps:

[0014] Centrifuge the sample to be tested, collect the supernatant, acidify it, centrifuge again, collect the supernatant, mix it with heavy water containing trisodium phosphate, centrifuge again, and collect the supernatant.

[0015] In one embodiment, the solvent used for acidification is trichloroacetic acid.

[0016] In one embodiment, the conditions for the first water bath heating include: a temperature of 35°C ± 1°C and a time of 30 min ± 10 min.

[0017] In one embodiment, the enzyme inactivation conditions include heating to 100°C ± 5°C.

[0018] In one embodiment, the multivariate statistical analysis includes one or more of PCA analysis, OPLS-DA analysis, and PLS-DA analysis.

[0019] In one embodiment, the high-field nuclear magnetic resonance detection further includes the following condition: the detection temperature is 25°C.

[0020] In one embodiment, the high-field nuclear magnetic resonance detection also includes the following condition: the number of sampling points is 32K.

[0021] In one embodiment, the high-field nuclear magnetic resonance detection further includes the following condition: the magnetic field strength is 600 MHz.

[0022] The present invention also provides the application of the bird's nest origin traceability method based on high-field nuclear magnetic resonance technology as described above in bird's nest quality control.

[0023] Compared with existing technologies, the bird's nest origin tracing method based on high-field nuclear magnetic resonance technology of the present invention has the following beneficial effects:

[0024] This invention employs high-field nuclear magnetic resonance (NMR) technology, while simultaneously limiting the spectral width, cumulative scan count, and extraction processing steps and parameters. Multivariate statistical analysis of the collected data allows for clear differentiation of bird's nests from different origins. The pretreatment steps required by the method provided by this invention are relatively simple and rapid, significantly shortening the detection time and demonstrating good stability. Furthermore, aside from the reagents used in sample preparation, almost no other consumables are required. The reagents used are relatively safe and environmentally friendly, with very small quantities required, making it environmentally friendly and posing minimal risk to human health, exhibiting good environmental tolerance.

[0025] In summary, this invention, as the first application of high-field nuclear magnetic resonance analysis technology to the traceability of bird's nest origins, is a highly efficient, accurate, convenient, environmentally friendly, and low-cost method that fills the gap in bird's nest origin traceability and expands the application scope of nuclear magnetic resonance technology. It is of great significance for the effective supervision of bird's nest quality in the future. Attached Figure Description

[0026] Figure 1 The multivariate analysis plot provided for this invention, wherein (a) represents PCA analysis and (b) represents PLS-DA analysis;

[0027] Figure 2 The OPLS-DA model diagram provided by the present invention is shown in which (a) represents the model of Vietnamese bird's nest and Malaysian bird's nest, and (b) represents the model of Indonesian bird's nest and Malaysian bird's nest. Detailed Implementation

[0028] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0029] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of the invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified. In the description of the present invention, "a number" means at least one, such as one, two, etc., unless otherwise explicitly specified.

[0030] The terms "preferred," "more preferably," etc., used in this invention refer to embodiments of the invention that provide certain beneficial effects under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are unavailable, nor is it intended to exclude other embodiments from the scope of this invention.

[0031] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0032] Unless otherwise specified, all percentages, fractions, and ratios are calculated based on the total mass of the compositions of the present invention. Unless otherwise specified, all masses of the listed ingredients are for the content of the active substance and therefore do not include solvents or byproducts that may be present in commercially available materials. The term "percentage by mass" is expressed using the symbol "%". Unless otherwise specified, all formulations and tests herein are performed at 25°C. The terms "comprising," "including," "containing," "containing," "having," or other variations thereof are intended to cover non-closed inclusions, and no distinction is made between these terms. The term "comprising" means additional steps and ingredients that may be added without affecting the final result. The compositions and methods / processes of the present invention comprise, consist of, and substantially consist of the essential elements and limitations described herein, as well as any additional or optional ingredients, components, steps, or limitations described herein. No distinction is made between the terms "efficacy," "performance," "effect," and "potency" herein.

[0033] The weights of the relevant components mentioned in the embodiments of this invention can refer not only to the specific content of each component, but also to the proportional relationship between the weights of the components. Therefore, any scaling up or down of the content of the relevant components according to the embodiments of this invention is within the scope disclosed in the embodiments of this invention. Specifically, the weights mentioned in the embodiments of this invention can be well-known mass units in the chemical industry, such as ug, mg, g, and kg.

[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0035] This invention provides a method for tracing the origin of bird's nests based on high-field nuclear magnetic resonance technology, comprising the following steps:

[0036] Bird's nests from different origins and the test substance were extracted, processed, and used to prepare the test samples;

[0037] The sample to be tested is pretreated, and then high-field nuclear magnetic resonance detection is performed to collect detection data.

[0038] Perform multivariate statistical analysis on the detection data;

[0039] The extraction process includes the following steps: soaking bird's nests from different origins and the test substance, mixing them separately with artificial saliva, heating in a water bath for the first time to prepare a first mixture; mixing the first mixture with artificial gastric juice, heating in a water bath for the second time, and then performing enzyme inactivation; the conditions for the second water bath heating include: temperature of 37℃±1℃ and time of 3h±0.5h.

[0040] High-field NMR detection includes the following conditions: spectral width of 12 kHz and cumulative scan count of 64.

[0041] Understandably, in this invention, the temperature of the second water bath heating includes, but is not limited to, 36°C, 37°C, and 38°C, and the time of the second water bath heating includes, but is not limited to, 2.5h, 2.6h, 2.7h, 2.8h, 2.9h, 3h, 3.1h, 3.2h, 3.3h, 3.4h, and 3.5h.

[0042] Understandably, in this invention, the spectral width of high-field nuclear magnetic resonance detection includes, but is not limited to, 10 kHz, 11 kHz, 12 kHz, 13 kHz, and 14 kHz.

[0043] Understandably, in this invention, soaking, also known as soaking or softening, involves immersing dried bird's nest in water to absorb water and expand, thereby improving the properties of the dried bird's nest, increasing its volume and softening its texture, thus meeting the requirements for stewing and cooking.

[0044] In a specific example, the pretreatment process includes the following steps:

[0045] Centrifuge the sample to be tested, collect the supernatant, acidify it, centrifuge again, collect the supernatant, mix it with heavy water containing trisodium phosphate, centrifuge again, and collect the supernatant.

[0046] In one specific example, the solvent used for acidification is trichloroacetic acid.

[0047] In a specific example, the conditions for the first water bath heating are: a temperature of 35℃±1℃ and a time of 30min±10min.

[0048] Understandably, in this invention, the temperature of the first water bath heating includes, but is not limited to, 34°C, 35°C, and 36°C, and the time of the first water bath heating includes, but is not limited to, 20 min, 21 min, 22 min, 23 min, 24 min, 25 min, 26 min, 27 min, 28 min, 29 min, 30 min, 31 min, 32 min, 33 min, 34 min, 35 min, 36 min, 37 min, 38 min, 39 min, and 40 min.

[0049] In a specific example, the conditions for enzyme inactivation include heating to 100℃±5℃.

[0050] In a specific example, multivariate statistical analysis includes one or more of PCA analysis, OPLS-DA analysis, and PLS-DA analysis.

[0051] In a specific example, high-field NMR detection also includes the following condition: the detection temperature is 25°C.

[0052] In a specific example, high-field NMR detection also includes the following condition: the number of sampling points is 32K.

[0053] In a specific example, high-field NMR detection also includes the following condition: magnetic field strength of 600 MHz.

[0054] In one specific example, the bird's nests from different origins are selected from at least two of the following: Vietnamese bird's nests, Malaysian bird's nests, and Indonesian bird's nests.

[0055] The present invention also provides the application of the above-mentioned method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology in the quality control of bird's nest.

[0056] The following detailed description of the bird's nest origin traceability method based on high-field nuclear magnetic resonance technology of the present invention is provided in conjunction with specific embodiments. Unless otherwise specified, all raw materials used in the following embodiments are commercially available products.

[0057] Example 1

[0058] This embodiment provides a method for tracing the origin of bird's nests based on high-field nuclear magnetic resonance technology, as detailed below:

[0059] 1. Experimental Instrument Parameters

[0060] A 600MHz Ascend nuclear magnetic resonance spectrometer was used for spectral acquisition. The instrument was equipped with an ultra-low temperature broadband multi-nuclear forward detection probe. The sampling pulse sequence was a zgpr sequence containing water peak suppression. The number of sampling points was set to 32K, the spectral width was set to 12kHz, the number of accumulations (NS, scan counts) was set to 64, and the experimental temperature was 298K.

[0061] 2. Sample preparation

[0062] Bird's nests (raw bird's nests) from Malaysia, Vietnam, and Indonesia were selected and divided into three groups based on their origin, with six samples per group. The bird's nests were first soaked at a solid-liquid ratio of 1:30. 250 mg (±5 mg) of the soaked bird's nest was then mixed with 250 μL of artificial saliva and heated in a water bath at 35°C for 30 minutes. Subsequently, 500 μL of artificial gastric juice was added to the mixture and shaken well. The mixture was then heated in a water bath at 37°C for 3 hours, followed by a temperature increase to 100°C to inactivate the relevant enzymes. The digested mixture was centrifuged (4℃, 10000rpm, 10min). 300µL of the supernatant was taken and mixed with trichloroacetic acid at a 1:1 ratio with shaking to remove protein from the bird's nest and prevent it from forming a complex with trisodium phosphate (TSP). Finally, the acidified solution was centrifuged again, and 300µL of the supernatant was taken. 300µL of D2O containing trisodium phosphate (TSP) was added, and the mixture was shaken and centrifuged again. 550µL of the supernatant was transferred to a 5mm NMR tube for HF-NMR detection.

[0063] 3. Multivariate statistical analysis

[0064] The results of multivariate statistical analysis of all test data are as follows: Figure 1 As shown in a), there are obvious intergroup distinctions in the first principal component of PCA for bird's nests from Malaysia (MY), Indonesia (ID), and Vietnam (VN); Figure 1 As shown in b), in the supervised PLS-DA (Partial Least Squares Regression Analysis) model, the bird's nests from the three origins show significant differences, suggesting that there are inherent differences between bird's nests from different origins. Further analysis using the OPLS-DA model, such as... Figure 2 As shown in a), R 2 X = 0.534, R 2 Y = 0.977, Q 2 Y = 0.829, as Figure 2 As shown in b), R 2 X = 0.577, R 2 Y = 0.936, Q 2 Y = 0.809, indicating that its R 2 X, R 2 Y, Q 2 Y values ​​greater than 0.5 indicate that the established OPLS-DA model is good. Figure 2 It can be seen that Malaysian bird's nests are significantly different from those from Vietnam and Indonesia.

[0065] Example 2

[0066] This embodiment provides a method for identifying adulterated bird's nest based on high-field nuclear magnetic resonance technology, as detailed below:

[0067] 1. Experimental Instrument Parameters

[0068] A 600MHz Ascend nuclear magnetic resonance spectrometer was used for spectral acquisition. The instrument was equipped with an ultra-low temperature broadband multi-nuclear forward detection probe. The sampling pulse sequence was a zgpr sequence containing water peak suppression. The number of sampling points was set to 32K, the spectral width was set to 12kHz, the number of accumulations (NS, scan counts) was set to 64, and the experimental temperature was 298K.

[0069] 2. Sample preparation

[0070] Take the test sample and soak it at a solid-liquid ratio of 1:30. Take 250 mg (±5 mg) of the soaked test sample and mix it with 250 uL of artificial saliva by shaking. Then heat it in a water bath at 35°C for 30 min. Subsequently, add 500 uL of artificial gastric juice to the mixture and shake it well. Heat it in a water bath at 37°C for 3 h. Then raise the temperature to 100°C to inactivate the relevant enzymes. The digested mixture was centrifuged (4℃, 10000rpm, 10min). 300µL of the supernatant was taken and mixed with trichloroacetic acid at a 1:1 ratio with shaking to remove protein from the bird's nest and prevent it from forming a complex with trisodium phosphate (TSP). Finally, the acidified solution was centrifuged again, and 300µL of the supernatant was taken. 300µL of D2O containing trisodium phosphate (TSP) was added, and the mixture was shaken and centrifuged again. 550µL of the supernatant was transferred to a 5mm NMR tube for HF-NMR detection.

[0071] 3. Multivariate statistical analysis

[0072] By substituting the test data into the PLS-DA (Partial Least Squares Regression Analysis) model and the OPLS-DA model established in Example 1, it can be determined whether the test substance is bird's nest from Malaysia, Vietnam, or Indonesia.

[0073] Comparative Example 1

[0074] This comparative example provides a method for tracing the origin of bird's nests based on high-field nuclear magnetic resonance technology, as detailed below:

[0075] 1. Experimental Instrument Parameters

[0076] A 600MHz Ascend nuclear magnetic resonance spectrometer was used for spectral acquisition. The instrument was equipped with an ultra-low temperature broadband multi-nuclear forward detection probe. The sampling pulse sequence was a zgpr sequence containing water peak suppression. The number of sampling points was set to 32K, the spectral width was set to 12kHz, the number of accumulations (NS, scan counts) was set to 64, and the experimental temperature was 298K.

[0077] 2. Sample preparation

[0078] Bird's nests (raw bird's nests) from Malaysia, Vietnam, and Indonesia were selected and divided into three groups based on their origin, with six samples in each group. The bird's nests were first soaked at a solid-liquid ratio of 1:30. 250 mg (±5 mg) of the soaked bird's nest was then mixed with 250 μL of artificial saliva and heated in a water bath at 35°C for 30 minutes. Subsequently, 500 μL of artificial gastric juice was added to the mixture and shaken well. The mixture was then heated in a water bath at 37°C for 1 hour, followed by a temperature increase to 100°C to inactivate the relevant enzymes. The digested mixture was centrifuged (4℃, 10000rpm, 10min). 300µL of the supernatant was taken and mixed with trichloroacetic acid at a 1:1 ratio with shaking to remove protein from the bird's nest and prevent it from forming a complex with trisodium phosphate (TSP). Finally, the acidified solution was centrifuged again, and 300µL of the supernatant was taken. 300µL of D2O containing trisodium phosphate (TSP) was added, and the mixture was shaken and centrifuged again. 550µL of the supernatant was transferred to a 5mm NMR tube for HF-NMR detection.

[0079] 3. Multivariate statistical analysis

[0080] The results of multivariate statistical analysis of all test data, using PCA first principal component and PLS-DA analysis, showed that bird's nests from Malaysia, Vietnam, and Indonesia could not be clearly distinguished. Analysis using the OPLS-DA model showed that R0... 2 X, R 2 Y, Q 2 Y < 0.5 indicates that the established OPLS-DA model lacks representativeness, therefore this method is not suitable for tracing the origin of bird's nest.

[0081] Comparative Example 2

[0082] This comparative example provides a method for identifying adulterated bird's nest based on high-field nuclear magnetic resonance technology, as detailed below:

[0083] 1. Experimental Instrument Parameters

[0084] A 600MHz Ascend nuclear magnetic resonance spectrometer was used for spectral acquisition. The instrument was equipped with an ultra-low temperature broadband multi-nuclear forward detection probe. The sampling pulse sequence was a zgpr sequence containing water peak suppression. The number of sampling points was set to 32K, the spectral width was set to 15kHz, the number of cumulative scans (NS) was set to 32, and the experimental temperature was 25℃.

[0085] 2. Sample preparation

[0086] Bird's nests (raw bird's nests) from Malaysia, Vietnam, and Indonesia were selected and divided into three groups based on their origin, with six samples per group. The bird's nests were first soaked at a solid-liquid ratio of 1:30. 250 mg (±5 mg) of the soaked bird's nest was then mixed with 250 μL of artificial saliva and heated in a water bath at 35°C for 30 minutes. Subsequently, 500 μL of artificial gastric juice was added to the mixture and shaken well. The mixture was then heated in a water bath at 37°C for 3 hours, followed by a temperature increase to 100°C to inactivate the relevant enzymes. The digested mixture was centrifuged (4℃, 10000rpm, 10min). 300µL of the supernatant was taken and mixed with trichloroacetic acid at a 1:1 ratio with shaking to remove protein from the bird's nest and prevent it from forming a complex with trisodium phosphate (TSP). Finally, the acidified solution was centrifuged again, and 300µL of the supernatant was taken. 300µL of D2O containing trisodium phosphate (TSP) was added, and the mixture was shaken and centrifuged again. 550µL of the supernatant was transferred to a 5mm NMR tube for HF-NMR detection.

[0087] 3. Multivariate statistical analysis

[0088] The results of multivariate statistical analysis of all test data, using PCA first principal component and PLS-DA analysis, showed that bird's nests from Malaysia, Vietnam, and Indonesia could not be clearly distinguished. Analysis using the OPLS-DA model showed that R0... 2 X, R 2 Y, Q 2 Y < 0.5 indicates that the established OPLS-DA model lacks representativeness, therefore this method is not suitable for tracing the origin of bird's nest.

[0089] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0090] The embodiments described above are merely illustrative of several implementations of the present invention, designed to facilitate a detailed understanding of the technical solutions of the present invention, but should not be construed as limiting the scope of protection of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the scope of protection of the present invention. It should be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided by the present invention through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this invention patent should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology, characterized in that, Includes the following steps: Bird's nests from different origins and the test substance were extracted, processed, and used to prepare the test samples; The sample to be tested is pretreated and then subjected to high-field nuclear magnetic resonance detection to collect detection data. The detection data were subjected to multivariate statistical analysis. The extraction process includes the following steps: soaking the bird's nests from different origins and the test substance, mixing them separately with artificial saliva, heating them in a water bath for the first time to prepare a first mixture; mixing the first mixture with artificial gastric juice, heating it in a water bath for the second time, and then performing enzyme inactivation; the conditions for the second water bath heating include: temperature of 37℃±1℃ and time of 3h±0.5h. High-field NMR detection includes the following conditions: the sampling pulse sequence is a zgpr sequence containing suppressed water peaks, the spectral width is 12kHz, and the cumulative number of scans is 64. The pretreatment process includes the following steps: Centrifuge the sample to be tested, take the supernatant, acidify it, centrifuge it again, take the supernatant, mix it with heavy water containing trisodium phosphate, centrifuge it again, and take the supernatant. The solvent used for acidification is trichloroacetic acid; The conditions for the first water bath heating are: temperature 35℃±1℃, time 30min±10min; The conditions for enzyme inactivation include: heating to 100℃±5℃; The bird's nests from different origins include raw bird's nests from Malaysia, Vietnam, and Indonesia.

2. The method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology according to claim 1, characterized in that, Multivariate statistical analysis includes one or more of PCA analysis, OPLS-DA analysis, and PLS-DA analysis.

3. The method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology according to any one of claims 1 to 2, characterized in that, High-field nuclear magnetic resonance (NMR) detection also includes the following conditions: the detection temperature is 25°C.

4. The method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology according to any one of claims 1 to 2, characterized in that, High-field NMR detection also includes the following conditions: the number of sampling points is 32K.

5. The method for tracing the origin of bird's nest based on high-field nuclear magnetic resonance technology according to any one of claims 1 to 2, characterized in that, High-field nuclear magnetic resonance detection also includes the following conditions: magnetic field strength of 600 MHz.

6. The application of the bird's nest origin traceability method based on high-field nuclear magnetic resonance technology as described in any one of claims 1 to 5 in the quality control of bird's nest.

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